WorldWideScience

Sample records for viable hlw vitrification

  1. HIGH LEVEL WASTE (HLW) VITRIFICATION EXPERIENCE IN THE US: APPLICATION OF GLASS PRODUCT/PROCESS CONTROL TO OTHERHLW AND HAZARDOUS WASTES

    Energy Technology Data Exchange (ETDEWEB)

    Jantzen, C; James Marra, J

    2007-09-17

    Vitrification is currently the most widely used technology for the treatment of high level radioactive wastes (HLW) throughout the world. At the Savannah River Site (SRS) actual HLW tank waste has successfully been processed to stringent product and process constraints without any rework into a stable borosilicate glass waste since 1996. A unique 'feed forward' statistical process control (SPC) has been used rather than statistical quality control (SQC). In SPC, the feed composition to the melter is controlled prior to vitrification. In SQC, the glass product is sampled after it is vitrified. Individual glass property models form the basis for the 'feed forward' SPC. The property models transform constraints on the melt and glass properties into constraints on the feed composition. The property models are mechanistic and depend on glass bonding/structure, thermodynamics, quasicrystalline melt species, and/or electron transfers. The mechanistic models have been validated over composition regions well outside of the regions for which they were developed because they are mechanistic. Mechanistic models allow accurate extension to radioactive and hazardous waste melts well outside the composition boundaries for which they were developed.

  2. FLUIDIZED BED STEAM REFORMING (FBSR) OF HIGH LEVEL WASTE (HLW) ORGANIC AND NITRATE DESTRUCTION PRIOR TO VITRIFICATION: CRUCIBLE SCALE TO ENGINEERING SCALE DEMONSTRATIONS AND NON-RADIOACTIVE TO RADIOACTIVE DEMONSTRATIONS

    Energy Technology Data Exchange (ETDEWEB)

    Jantzen, C; Michael Williams, M; Gene Daniel, G; Paul Burket, P; Charles Crawford, C

    2009-02-07

    Over a decade ago, an in-tank precipitation process to remove Cs-137 from radioactive high level waste (HLW) supernates was demonstrated at the Savannah River Site (SRS). The full scale demonstration with actual HLW was performed in SRS Tank 48 (T48). Sodium tetraphenylborate (NaTPB) was added to enable Cs-137 extraction as CsTPB. The CsTPB, an organic, and its decomposition products proved to be problematic for subsequent processing of the Cs-137 precipitate in the SRS HLW vitrification facility for ultimate disposal in a HLW repository. Fluidized Bed Steam Reforming (FBSR) is being considered as a technology for destroying the organics and nitrates in the T48 waste to render it compatible with subsequent HLW vitrification. During FBSR processing the T48 waste is converted into organic-free and nitrate-free carbonate-based minerals which are water soluble. The soluble nature of the carbonate-based minerals allows them to be dissolved and pumped to the vitrification facility or returned to the tank farm for future vitrification. The initial use of the FBSR process for T48 waste was demonstrated with simulated waste in 2003 at the Savannah River National Laboratory (SRNL) using a specially designed sealed crucible test that reproduces the FBSR pyrolysis reactions, i.e. carbonate formation, organic and nitrate destruction. This was followed by pilot scale testing of simulants at the Science Applications International Corporation (SAIC) Science & Technology Application Research (STAR) Center in Idaho Falls, ID by Idaho National Laboratory (INL) and SRNL in 2003-4 and then engineering scale demonstrations by THOR{reg_sign} Treatment Technologies (TTT) and SRS/SRNL at the Hazen Research, Inc. (HRI) test facility in Golden, CO in 2006 and 2008. Radioactive sealed crucible testing with real T48 waste was performed at SRNL in 2008, and radioactive Benchscale Steam Reformer (BSR) testing was performed in the SRNL Shielded Cell Facility (SCF) in 2008.

  3. Chemical Oxygen Demand (COD) For Monitoring Reduction-Oxidation (Redox) Equilibrium During High Level Waste (HLW) Vitrification

    Energy Technology Data Exchange (ETDEWEB)

    JANTZEN, CAROLM.

    2004-04-30

    High-level nuclear waste is being immobilized at the Savannah River Site by vitrification into borosilicate glass at the Defense Waste Processing Facility. Control of the REDuction/OXidation (REDOX) equilibrium in the DWPF melter is critical for processing high level liquid wastes. Based upon previous research, an acceptable iron REDOX ratio was defined for the DWPF melts as 0.09 Fe2/SFe 0.33. Controlling the DWPF melter at a REDuction/OXidation (REDOX) equilibrium ofFe2/SFe 0.33 prevents the potential for metallic and metallic sulfide species to form and accumulate on the floor of the melter. Control of foaming due to deoxygenation of manganic species is achieved by converting 66-100 of the MnO2 or Mn2O3 species in a waste feed to MnO before the waste is fed to the DWPF melter. At the lower redox limit of Fe 2/SFe 0.09 about 99 of the Mn 4/Mn 3 is converted to Mn 2. Therefore, the lower REDOX limit eliminates melter foaming from deoxygenation. Organic and nitrate concentrations in the DWPF melter feed are the major parameters influencing melt REDOX. Organics such as formates act as reductants while nitrates, nitrites, and manganic (Mn 4 and Mn 3) species act as oxidants. During melting, the REDOX of the melt pool cannot be measured. Therefore, the Fe 2/SFe ratio in the glass poured from the melter must be related to melter feed organic and oxidant concentrations to ensure production of a high quality glass without impacting production rate (e.g., from foaming) or melter life (e.g., from metal formation and accumulation).

  4. MECHANISMS OF PHASE FORMATION IN THE VITRIFICATION OF HIGH-FERROUS SAVANNAH RIVER SITE SB2 HLW SLUDGE SURROGATE - 9300

    Energy Technology Data Exchange (ETDEWEB)

    Marra, J

    2008-08-27

    Phase formation mechanisms associated with the vitrification of high-ferrous Savannah River Site (SRS) Sludge Batch 2 (SB2) high level waste surrogate were studied by infrared spectroscopy (IRS) and X-ray diffraction (XRD). Two mixtures at 50 wt% waste loading with commercially available Frit 320 (Li{sub 2}O - 8 wt %, B{sub 2}O{sub 3} - 8 wt %, Na{sub 2}O - 12 wt %, SiO{sub 2} - 72 wt %) and batch chemicals (LiOH {center_dot} H{sub 2}O, H{sub 3}BO{sub 3}, NaNO{sub 3}, SiO{sub 2}) to represent the frit formulation were prepared as slurries with a water content of {approx}50 wt%. The mixtures were air-dried at a temperature of 115 C and heat-treated at 500, 700, 900, 1000, 1100, 1200, and 1300 C for 1 hr at each temperature. Infrared spectra and XRD patterns of the products produced at each temperature were recorded. In both mixtures prepared using frit and batch chemicals to represent the frit, phase formation reactions were completed within the temperature range between 900 and 1000 C. However, residual quartz was still present in glass produced from the mixture with batch chemicals even at 1100 C. Although, the phase composition and structure of the glassy products obtained from both mixtures at temperatures over 1000 C were similar, the products obtained from the mixture using actual frit were more homogeneous than those from the mixture with batch chemicals. Thus, the use of frit rather than batch chemicals reduced the temperature range of phase formation and provided for production of higher quality glass.

  5. HLW Tank Space Management, Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Miller, M.S.; Abell, G.; Garrett, R.; d' Entremont, P.; Fowler, J.R.; Mahoney, M.; Poe, L.

    1999-09-20

    The HLW Tank Space Management Team (SM Team) was chartered to select and recommend an HLW Tank Space Management Strategy (Strategy) for the HLW Management Division of Westinghouse Savannah River Co. (WSRC) until an alternative salt disposition process is operational. Because the alternative salt disposition process will not be available to remove soluble radionuclides in HLW until 2009, the selected Strategy must assure that it safely receives and stores HLW at least until 2009 while continuing to supply sludge slurry to the DWPF vitrification process.

  6. FINAL REPORT MELTER TESTS WITH AZ-101 HLW SIMULANT USING A DURAMELTER 100 VITRIFICATION SYSTEM VSL-01R10N0-1 REV 1 2/25/02

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; KOT WK; PEGG IL

    2011-12-29

    This report provides data, analyses, and conclusions from a series of tests that were conducted at the Vitreous State Laboratory of The Catholic of America (VSL) to determine the processing rates that are achievable with AZ-101 HLW simulants and corresponding melter feeds on a DuraMelter 100 (DM100) vitrification system. One of the most critical pieces of information in determining the required size of the RPP-WTP HLW melter is the specific glass production rate in terms of the mass of glass that can be produced per unit area of melt surface per unit time. The specific glass production rate together with the waste loading (essentially, the ratio of waste-in to glass-out, which is determined from glass formulation activities) determines the melt area that is needed to achieve a given waste processing rate with due allowance for system availability. Tests conducted during Part B1 (VSL-00R2590-2) on the DM1000 vitrification system installed at the Vitreous State Laboratory of The Catholic University of America showed that, without the use of bubblers, glass production rates with AZ-101 and C-106/AY-102 simulants were significantly lower than the Project design basis rate of 0.4 MT/m{sup 2}/d. Conversely, three-fold increases over the design basis rate were demonstrated with the use of bubblers. Furthermore, an un-bubbled control test using a replica of the melter feed used in cold commissioning tests at West Valley reproduced the rates that were observed with that feed on the WVDP production melter. More recent tests conducted on the DM1200 system, which more closely represents the present RPP-WTP design, are in general agreement with these earlier results. Screening tests conducted on the DM10 system have provided good indications of the larger-scale processing rates with bubblers (for both HL W and LAW feeds) but significantly overestimated the DM1000 un-bubbled rate observed for C-106/AY-102 melter feeds. This behavior is believed to be a consequence of the role of

  7. Final Report - Effects of High Spinel and Chromium Oxide Crystal Contents on Simulated HLW Vitrification in DM100 Melter Tests, VSL-09R1520-1, Rev. 0, dated 6/22/09

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Matlack, K. S.; Kot, W.; Pegg, I. L.; Chaudhuri, M.; Lutze, W.

    2013-11-13

    The principal objective of the work was to evaluate the effects of spinel and chromium oxide particles on WTP HLW melter operations and potential impacts on melter life. This was accomplished through a combination of crucible-scale tests, settling and rheological tests, and tests on the DM100 melter system. Crucible testing was designed to develop and identify HLW glass compositions with high waste loadings that exhibit formation of crystalline spinel and/or chromium oxide phases up to relatively high crystal contents (i.e., > 1 vol%). Characterization of crystal settling and the effects on melt rheology was performed on the HLW glass formulations. Appropriate candidate HLW glass formulations were selected, based on characterization results, to support subsequent melter tests. In the present work, crucible melts were formulated that exhibit up to about 4.4 vol% crystallization.

  8. FINAL REPORT DETERMINATION OF THE PROCESSING RATE OF RPP WTP HLW SIMULANTS USING A DURAMELTER J 1000 VITRIFICATION SYSTEM VSL-00R2590-2 REV 0 8/21/00

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; KOT WK; PEREZ-CARDENAS F; PEGG IL

    2011-12-29

    This report provides data, analysis, and conclusions from a series of tests that were conducted at the Vitreous State Laboratory of The Catholic University of America (VSL) to determine the melter processing rates that are achievable with RPP-WTP HLW simulants. The principal findings were presented earlier in a summary report (VSL-00R2S90-l) but the present report provides additional details. One of the most critical pieces of information in determining the required size of the RPP-WTP HLW melter is the specific glass production rate in terms of the mass of glass that can be produced per unit area of melt surface per unit time. The specific glass production rate together with the waste loading (essentially, the ratio of waste-in to glass-out, which is determined from glass formulation activities) determines the melt area that is needed to achieve a given waste processing rate with due allowance for system availability. As a consequence of the limited amount of relevant information, there exists, for good reasons, a significant disparity between design-base specific glass production rates for the RPP-WTP LAW and HLW conceptual designs (1.0 MT/m{sup 2}/d and 0.4 MT/m{sup 2}/d, respectively); furthermore, small-scale melter tests with HLW simulants that were conducted during Part A indicated typical processing rates with bubbling of around 2.0 MT/m{sup 2}/d. This range translates into more than a factor of five variation in the resultant surface area of the HLW melter, which is clearly not without significant consequence. It is clear that an undersized melter is undesirable in that it will not be able to support the required waste processing rates. It is less obvious that there are potential disadvantages associated with an oversized melter, over and above the increased capital costs. A melt surface that is consistently underutilized will have poor cold cap coverage, which will result in increased volatilization from the melt (which is generally undesirable) and

  9. DEVELOPMENT OF GLASS MATRICES FOR HLW RADIOACTIVE WASTES

    Energy Technology Data Exchange (ETDEWEB)

    Jantzen, C.

    2010-03-18

    Vitrification is currently the most widely used technology for the treatment of high level radioactive wastes (HLW) throughout the world. Most of the nations that have generated HLW are immobilizing in either borosilicate glass or phosphate glass. One of the primary reasons that glass has become the most widely used immobilization media is the relative simplicity of the vitrification process, e.g. melt waste plus glass forming frit additives and cast. A second reason that glass has become widely used for HLW is that the short range order (SRO) and medium range order (MRO) found in glass atomistically bonds the radionuclides and governs the melt properties such as viscosity, resistivity, sulphate solubility. The molecular structure of glass controls contaminant/radionuclide release by establishing the distribution of ion exchange sites, hydrolysis sites, and the access of water to those sites. The molecular structure is flexible and hence accounts for the flexibility of glass formulations to waste variability. Nuclear waste glasses melt between 1050-1150 C which minimizes the volatility of radioactive components such as Tc{sup 99}, Cs{sup 137}, and I{sup 129}. Nuclear waste glasses have good long term stability including irradiation resistance. Process control models based on the molecular structure of glass have been mechanistically derived and have been demonstrated to be accurate enough to control the world's largest HLW Joule heated ceramic melter in the US since 1996 at 95% confidence.

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

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Wang, C.; Gan, H.; Pegg, I. L.; Chaudhuri, M.; Kot, W.; Feng, Z.; Viragh, C.; McKeown, D. A.; Joseph, I.; Muller, I. S.; Cecil, R.; Zhao, W.

    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 melters 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

  11. Support for HLW Direct Feed - Phase 2, VSL-15R3440-1

    Energy Technology Data Exchange (ETDEWEB)

    Matlack, K. S. [The Catholic Univ. of America, Washington, DC (United States); Pegg, I. [The Catholic Univ. of America, Washington, DC (United States); Joseph, I. [EnergySolutions, Columbia, MD (United States); Kot, W. K. [The Catholic Univ. of America, Washington, DC (United States)

    2017-03-20

    This report describes work performed to develop and test new glass and feed formulations originating from a potential flow-sheet for the direct vitrification of High Level Waste (HLW) with minimal or no pretreatment. In the HLW direct feed option that is under consideration for early operations at the Hanford Tank Waste Treatment and Immobilization Plant (WTP), the pretreatment facility would be bypassed in order to support an earlier start-up of the vitrification facility. For HLW, this would mean that the ultrafiltration and caustic leaching operations that would otherwise have been performed in the pretreatment facility would either not be performed or would be replaced by an interim pretreatment function (in-tank leaching and settling, for example). These changes would likely affect glass formulations and waste loadings and have impacts on the downstream vitrification operations. Modification of the pretreatment process may result in: (i) Higher aluminum contents if caustic leaching is not performed; (ii) Higher chromium contents if oxidative leaching is not performed; (iii) A higher fraction of supernate in the HLW feed resulting from the lower efficiency of in-tank washing; and (iv) A higher water content due to the likely lower effectiveness of in-tank settling compared to ultrafiltration. The HLW direct feed option has also been proposed as a potential route for treating HLW streams that contain the highest concentrations of fast-settling plutoniumcontaining particles, thereby avoiding some of the potential issues associated with such particles in the WTP Pretreatment facility [1]. In response, the work presented herein focuses on the impacts of increased supernate and water content on wastes from one of the candidate source tanks for the direct feed option that is high in plutonium.

  12. HLW Disposal System Development

    Energy Technology Data Exchange (ETDEWEB)

    Choi, J. W.; Choi, H. J.; Lee, J. Y. (and others)

    2007-06-15

    A KRS is suggested through design requirement analysis of the buffer and the canister which are the constituent of disposal system engineered barrier and HLW management plans are proposed. In the aspect of radionuclide retention capacity, the thickness of the buffer is determined 0.5m, the shape to be disc and ring and the dry density to be 1.6 g/cm{sup 3}. The maximum temperature of the buffer is below 100 .deg. which meets the design requirement. And bentonite blocks with 5 wt% of graphite showed more than 1.0 W/mK of thermal conductivity without the addition of sand. The result of the thermal analysis for proposed double-layered buffer shows that decrease of 7 .deg. C in maximum temperature of the buffer. For the disposal canister, the copper for the outer shell material and cast iron for the inner structure material is recommended considering the results analyzed in terms of performance of the canisters and manufacturability and the geochemical properties of deep groundwater sampled from the research area with granite, salt water intrusion, and the heavy weight of the canister. The results of safety analysis for the canister shows that the criticality for the normal case including uncertainty is the value of 0.816 which meets subcritical condition. Considering nation's 'Basic Plan for Electric Power Demand and Supply' and based on the scenario of disposing CANDU spent fuels in the first phase, the disposal system that the repository will be excavated in eight phases with the construction of the Underground Research Laboratory (URL) beginning in 2020 and commissioning in 2040 until the closure of the repository is proposed. Since there is close correlation between domestic HLW management plans and front-end/back-end fuel cycle plans causing such a great sensitivity of international environment factor, items related to assuring the non-proliferation and observing the international standard are showed to be the influential factor and acceptability

  13. PAIRWISE BLENDING OF HIGH LEVEL WASTE (HLW)

    Energy Technology Data Exchange (ETDEWEB)

    CERTA, P.J.

    2006-02-22

    The primary objective of this study is to demonstrate a mission scenario that uses pairwise and incidental blending of high level waste (HLW) to reduce the total mass of HLW glass. Secondary objectives include understanding how recent refinements to the tank waste inventory and solubility assumptions affect the mass of HLW glass and how logistical constraints may affect the efficacy of HLW blending.

  14. Vitrification Facility integrated system performance testing report

    Energy Technology Data Exchange (ETDEWEB)

    Elliott, D.

    1997-05-01

    This report provides a summary of component and system performance testing associated with the Vitrification Facility (VF) following construction turnover. The VF at the West Valley Demonstration Project (WVDP) was designed to convert stored radioactive waste into a stable glass form for eventual disposal in a federal repository. Following an initial Functional and Checkout Testing of Systems (FACTS) Program and subsequent conversion of test stand equipment into the final VF, a testing program was executed to demonstrate successful performance of the components, subsystems, and systems that make up the vitrification process. Systems were started up and brought on line as construction was completed, until integrated system operation could be demonstrated to produce borosilicate glass using nonradioactive waste simulant. Integrated system testing and operation culminated with a successful Operational Readiness Review (ORR) and Department of Energy (DOE) approval to initiate vitrification of high-level waste (HLW) on June 19, 1996. Performance and integrated operational test runs conducted during the test program provided a means for critical examination, observation, and evaluation of the vitrification system. Test data taken for each Test Instruction Procedure (TIP) was used to evaluate component performance against system design and acceptance criteria, while test observations were used to correct, modify, or improve system operation. This process was critical in establishing operating conditions for the entire vitrification process.

  15. Development of the high-level waste high-temperature melter feed preparation flowsheet for vitrification process testing

    Energy Technology Data Exchange (ETDEWEB)

    Seymour, R.G.

    1995-02-17

    High-level waste (HLW) feed preparation flowsheet development was initiated in fiscal year (FY) 1994 to evaluate alternative flowsheets for preparing melter feed for high-temperature melter (HTM) vitrification testing. Three flowsheets were proposed that might lead to increased processing capacity relative to the Hanford Waste Vitrification Plant (HWVP) and that were flexible enough to use with other HLW melter technologies. This document describes the decision path that led to the selection of flowsheets to be tested in the FY 1994 small-scale HTM tests. Feed preparation flowsheet development for the HLW HTM was based on the feed preparation flowsheet that was developed for the HWVP. This approach allowed the HLW program to build upon the extensive feed preparation flowsheet database developed under the HWVP Project. Primary adjustments to the HWVP flowsheet were to the acid adjustment and glass component additions. Developmental background regarding the individual features of the HLW feed preparation flowsheets is provided. Applicability of the HWVP flowsheet features to the new HLW vitrification mission is discussed. The proposed flowsheets were tested at the laboratory-scale at Pacific Northwest Laboratory. Based on the results of this testing and previously established criteria, a reductant-based flowsheet using glycolic acid and a nitric acid-based flowsheet were selected for the FY 1994 small-scale HTM testing.

  16. The Effect of Vitrification on Follicular Morphology of Ovarian Rat

    Directory of Open Access Journals (Sweden)

    Foroozan Esmaeilzadeh

    2015-08-01

    Full Text Available Background & Objective: Some efforts have been made for keeping cryopreservation of gametes and embryos safe, including new vitrification methods of the ovary. This study evaluates the effect of ethylene glycole vitrification on follicular morphology of ovarian rat.Materials & Methods: Eighty ovaries belonging to 40 rats are divided into 2 groups. Twenty five ovaries are control group, 25 the vitrification, and30 toxicologic effects. For freezing, equilibrium solution, ethylene glycole and methyl sulfoxide are used. For defreezeing, different concentrations of saccharose and for morphological evaluation, H&E staining are undertaken. The number of healthy and atretic follicles are determined after 24 hours, 1 week and one month after vitrification.Results: No morphological changes are observed in all follicular cells. The percent of primordial, primary, secondary, anthral and developed follicles in the vitrification group are 34.5%, 17.7%, 17.4%, 15.2% and 50.3%. In vitrification and toxicological groups, the percent of both normal and atretic follicles is 47.5% and 11.9%. These figures for the control group were 59.7% and 16.9%. In vitrification method, 91% of oocytes are viable, 81% have mitosis, and 50% enters blastocyst stage.Conclusion: Because in vitrification of ovary in comparison with the follicles, many types of follicles in different cycles can be recovered with no morphological and structural changes, vitrification of ovary can be a safe method for cryopreservation of the oocytes

  17. Defense HLW Glass Degradation Model

    Energy Technology Data Exchange (ETDEWEB)

    D. Strachan

    2004-10-20

    The purpose of this report is to document the development of a model for calculating the release rate for radionuclides and other key elements from high-level radioactive waste (HLW) glasses under exposure conditions relevant to the performance of the repository. Several glass compositions are planned for the repository, some of which have yet to be identified (i.e., glasses from Hanford and Idaho National Engineering and Environmental Laboratory). The mechanism for glass dissolution is the same for these glasses and the glasses yet to be developed for the disposal of DOE wastes. All of these glasses will be of a quality consistent with the glasses used to develop this report.

  18. Demonstrating compliance with WAPS 1.3 in the Hanford waste vitrification plant process

    Energy Technology Data Exchange (ETDEWEB)

    Bryan, M.F.; Piepel, G.F.; Simpson, D.B.

    1996-03-01

    The high-level waste (HLW) vitrification plant at the Hanford Site was being designed to immobilize transuranic and high-level radioactive waste in borosilicate glass. This document describes the statistical procedure to be used in verifying compliance with requirements imposed by Section 1.3 of the Waste Acceptance Product Specifications (WAPS, USDOE 1993). WAPS 1.3 is a specification for ``product consistency,`` as measured by the Product Consistency Test (PCT, Jantzen 1992b), for each of three elements: lithium, sodium, and boron. Properties of a process batch and the resulting glass are largely determined by the composition of the feed material. Empirical models are being developed to estimate some property values, including PCT results, from data on feed composition. These models will be used in conjunction with measurements of feed composition to control the HLW vitrification process and product.

  19. Korean Reference HLW Disposal System

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Heui Joo; Lee, J. Y.; Kim, S. S. (and others)

    2008-03-15

    This report outlines the results related to the development of Korean Reference Disposal System for High-level radioactive wastes. The research has been supported around for 10 years through a long-term research plan by MOST. The reference disposal method was selected via the first stage of the research during which the technical guidelines for the geological disposal of HLW were determined too. At the second stage of the research, the conceptual design of the reference disposal system was made. For this purpose the characteristics of the reference spent fuels from PWR and CANDU reactors were specified, and the material and specifications of the canisters were determined in term of structural analysis and manufacturing capability in Korea. Also, the mechanical and chemical characteristics of the domestic Ca-bentonite were analyzed in order to supply the basic design parameters of the buffer. Based on these parameters the thermal and mechanical analysis of the near-field was carried out. Thermal-Hydraulic-Mechanical behavior of the disposal system was analyzed. The reference disposal system was proposed through the second year research. At the final third stage of the research, the Korean Reference disposal System including the engineered barrier, surface facilities, and underground facilities was proposed through the performance analysis of the disposal system.

  20. FINAL REPORT TESTS ON THE DURAMELTER 1200 HLW PILOT MELTER SYSTEM USING AZ-101 HLW SIMULANTS VSL-02R0100-2 REV 1 2/17/03

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; KOT WK; BARDAKCI T; GONG W; D' ANGELO NA; SCHATZ TR; PEGG IL

    2011-12-29

    This document provides the final report on data and results obtained from a series of nine tests performed on the one-third scale DuraMelter{trademark} 1200 (DM1200) HLW Pilot Melter system that has been installed at VSL with an integrated prototypical off-gas treatment system. That system has replaced the DM1000 system that was used for HLW throughput testing during Part B1 [1]. Both melters have similar melt surface areas (1.2 m{sup 2}) but the DM1200 is prototypical of the present RPP-WTP HLW melter design whereas the DM1000 was not. These tests were performed under a corresponding RPP-WTP Test Specification and associated Test Plans. The nine tests reported here were preceded by an initial series of short-duration tests conducted to support the start-up and commissioning of this system. This report is a followup to the previously issued Preliminary Data Summary Reports. The DM1200 system was deployed for testing and confirmation of basic design, operability, flow sheet, and process control assumptions as well as for support of waste form qualification and permitting. These tests include data on processing rates, off-gas treatment system performance, recycle stream compositions, as well as process operability and reliability. Consequently, this system is a key component of the overall HLW vitrification development strategy. The primary objective of the present series of tests was to determine the effects of a variety of parameters on the glass production rate in comparison to the RPP-WTP HL W design basis of 400 kg/m{sup 2}/d. Previous testing on the DMIOOO system [1] concluded that achievement of that rate with simulants of projected WTP melter feeds (AZ-101 and C-106/AY-102) was unlikely without the use of bubblers. As part of those tests, the same feed that was used during the cold-commissioning of the West Valley Demonstration Project (WVDP) HLW vitrification system was run on the DM1000 system. The DM1000 tests reproduced the rates that were obtained at the

  1. Glass matrices for vitrification of radioactive waste - an Update on R & D Efforts

    Science.gov (United States)

    Raj, Kanwar; Kaushik, C. P.

    2009-07-01

    Radioactive waste gets generated at different stages of nuclear fuel cycle like mining/milling, fuel fabrication, reactor operation, reprocessing of spent fuel and the production & application of radioisotopes for various industrial, medical and research purposes. High Level radioactive Waste (HLW) is generated during reprocessing of spent nuclear fuel and it contains most of the radioactivity present in entire fuel cycle. Vitrification of HLW in borosilicate matrix is being practiced using induction heated metallic melters at industrial scale plants at Tarapur and Trombay [1]. The nature of HLW largely depends on off - reactor cooling of spent nuclear fuel, its type and burn - up, and reprocessing flow sheet. In view of varying characteristics, processing of HLW at Tarapur and Trombay has offered a wide spectrum of challenges in terms of development of matrices and characterization to accommodate compositional changes in waste. The present paper summarizes details of extensive R and D efforts made in the Department of Atomic Energy towards development and characterization of glass formulations for immobilization of HLW.

  2. Final Report - Melt Rate Enhancement for High Aluminum HLW Glass Formulation, VSL-08R1360-1, Rev. 0, dated 12/19/08

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Pegg, I. L.; Chaudhuri, M.; Gong, W.; Gan, H.; Matlack, K. S.; Bardakci, T.; Kot, W.

    2013-11-13

    The principal objective of the work reported here was to develop and identify HLW glass compositions that maximize waste processing rates for the aluminum limted waste composition specified by ORP while maintaining high waste loadings and acceptable glass properties. This was accomplished through a combination of crucible-scale tests, confirmation tests on the DM100 melter system, and demonstration at pilot scale (DM1200). The DM100-BL unit was selected for these tests since it was used previously with the HLW waste streams evaluated in this study, was used for tests on HLW glass compositions to support subsequent tests on the HLW Pilot Melter, conduct tests to determine the effect of various glass properties (viscosity and conductivity) and oxide concentrations on glass production rates with HLW feed streams, and to assess the volatility of cesium and technetium during the vitrification of an HLW AZ-102 composition. The same melter was selected for the present tests in order to maintain comparisons between the previously collected data. These tests provide information on melter processing characteristics and off-gas data, including formation of secondary phases and partitioning. Once DM100 tests were completed, one of the compositions was selected for further testing on the DM1200; the DM1200 system has been used for processing a variety of simulated Hanford waste streams. Tests on the larger melter provide processing data at one third of the scale of the actual WTP HLW melter and, therefore, provide a more accurate and reliable assessment of production rates and potential processing issues. The work focused on maximizing waste processing rates for high aluminum HLW compositions. In view of the diversity of forms of aluminum in the Hanford tanks, tests were also conducted on the DM100 to determine the effect of changes in the form of aluminum on feed properties and production rate. In addition, the work evaluated the effect on production rate of modest increases

  3. Tank Waste Remediation System tank waste pretreatment and vitrification process development testing requirements assessment

    Energy Technology Data Exchange (ETDEWEB)

    Howden, G.F.

    1994-10-24

    A multi-faceted study was initiated in November 1993 to provide assurance that needed testing capabilities, facilities, and support infrastructure (sampling systems, casks, transportation systems, permits, etc.) would be available when needed for process and equipment development to support pretreatment and vitrification facility design and construction schedules. This first major report provides a snapshot of the known testing needs for pretreatment, low-level waste (LLW) and high-level waste (HLW) vitrification, and documents the results of a series of preliminary studies and workshops to define the issues needing resolution by cold or hot testing. Identified in this report are more than 140 Hanford Site tank waste pretreatment and LLW/HLW vitrification technology issues that can only be resolved by testing. The report also broadly characterizes the level of testing needed to resolve each issue. A second report will provide a strategy(ies) for ensuring timely test capability. Later reports will assess the capabilities of existing facilities to support needed testing and will recommend siting of the tests together with needed facility and infrastructure upgrades or additions.

  4. FINAL REPORT START-UP AND COMMISSIONING TESTS ON THE DURAMELTER 1200 HLW PILOT MELTER SYSTEM USING AZ-101 HLW SIMULANTS VSL-01R0100-2 REV 0 1/20/03

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; KOT WK; BRANDYS M; WILSON CN; SCHATZ TR; GONG W; PEGG IL

    2011-12-29

    This document provides the final report on data and results obtained from commissioning tests performed on the one-third scale DuraMelter{trademark} 1200 (DM 1200) HLW Pilot Melter system that has been installed at VSL with an integrated prototypical off-gas treatment system. That system has replaced the DM1000 system that was used for HLW throughput testing during Part BI [1]. Both melters have similar melt surface areas (1.2 m{sup 2}) but the DM1200 is prototypical of the present RPP-WTP HLW melter design whereas the DM1000 was not. These tests were performed under a corresponding RPP-WTP Test Specification and associated Test Plan. This report is a followup to the previously issued Preliminary Data Summary Report. The DM1200 system will be used for testing and confirmation of basic design, operability, flow sheet, and process control assumptions as well as for support of waste form qualification and permitting. This will include data on processing rates, off-gas treatment system performance, recycle stream compositions, as well as process operability and reliability. Consequently, this system is a key component of the overall HLW vitrification development strategy. The results presented in this report are from the initial series of short-duration tests that were conducted to support the start-up and commissioning of this system prior to conducting the main body of development tests that have been planned for this system. These tests were directed primarily at system 'debugging,' operator training, and procedure refinement. The AZ-101 waste simulant and glass composition that was used for previous testing was selected for these tests.

  5. Vitrification publication bibliography

    Energy Technology Data Exchange (ETDEWEB)

    Schmieman, E.; Johns, W.E.

    1996-02-01

    This document was compiled by a group of about 12 graduate students in the Department of Mechanical Engineering and Material Science at Washington State University and was funded by the U.S. Department of Energy. The literature search resulting in the compilation of this bibliography was designed to be an exhaustive search for research and development work involving the vitrification of mixed wastes, published by domestic and foreign researchers, primarily during 1989-1994. The search techniques were dominated by electronic methods and this bibliography is also available in electronic format, Windows Reference Manager.

  6. COMSOL Multiphysics Model for HLW Canister Filling

    Energy Technology Data Exchange (ETDEWEB)

    Kesterson, M. R. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2016-04-11

    The U.S. Department of Energy (DOE) is building a Tank Waste Treatment and Immobilization Plant (WTP) at the Hanford Site in Washington to remediate 55 million gallons of radioactive waste that is being temporarily stored in 177 underground tanks. Efforts are being made to increase the loading of Hanford tank wastes in glass while meeting melter lifetime expectancies and process, regulatory, and product quality requirements. Wastes containing high concentrations of Al2O3 and Na2O can contribute to nepheline (generally NaAlSiO4) crystallization, which can sharply reduce the chemical durability of high level waste (HLW) glass. Nepheline crystallization can occur during slow cooling of the glass within the stainless steel canister. The purpose of this work was to develop a model that can be used to predict temperatures of the glass in a WTP HLW canister during filling and cooling. The intent of the model is to support scoping work in the laboratory. It is not intended to provide precise predictions of temperature profiles, but rather to provide a simplified representation of glass cooling profiles within a full scale, WTP HLW canister under various glass pouring rates. These data will be used to support laboratory studies for an improved understanding of the mechanisms of nepheline crystallization. The model was created using COMSOL Multiphysics, a commercially available software. The model results were compared to available experimental data, TRR-PLT-080, and were found to yield sufficient results for the scoping nature of the study. The simulated temperatures were within 60 ºC for the centerline, 0.0762m (3 inch) from centerline, and 0.2286m (9 inch) from centerline thermocouples once the thermocouples were covered with glass. The temperature difference between the experimental and simulated values reduced to 40 ºC, 4 hours after the thermocouple was covered, and down to 20 ºC, 6 hours after the thermocouple was covered

  7. Development And Initial Testing Of Off-Gas Recycle Liquid From The WTP Low Activity Waste Vitrification Process - 14333

    Energy Technology Data Exchange (ETDEWEB)

    McCabe, Daniel J.; Wilmarth, William R.; Nash, Charles A.; Taylor-Pashow, Kathryn M.; Adamson, Duane J.; Crawford, Charles L.; Morse, Megan M.

    2014-01-07

    The Waste Treatment and Immobilization Plant (WTP) process flow was designed to pre-treat feed from the Hanford tank farms, separate it into a High Level Waste (HLW) and Low Activity Waste (LAW) fraction and vitrify each fraction in separate facilities. Vitrification of the waste generates an aqueous condensate stream from the off-gas processes. This stream originates from two off-gas treatment unit operations, the Submerged Bed Scrubber (SBS) and the Wet Electrospray Precipitator (WESP). Currently, the baseline plan for disposition of the stream from the LAW melter is to recycle it to the Pretreatment facility where it gets evaporated and processed into the LAW melter again. If the Pretreatment facility is not available, the baseline disposition pathway is not viable. Additionally, some components in the stream are volatile at melter temperatures, thereby accumulating to high concentrations in the scrubbed stream. It would be highly beneficial to divert this stream to an alternate disposition path to alleviate the close-coupled operation of the LAW vitrification and Pretreatment facilities, and to improve long-term throughput and efficiency of the WTP system. In order to determine an alternate disposition path for the LAW SBS/WESP Recycle stream, a range of options are being studied. A simulant of the LAW Off-Gas Condensate was developed, based on the projected composition of this stream, and comparison with pilot-scale testing. The primary radionuclide that vaporizes and accumulates in the stream is Tc-99, but small amounts of several other radionuclides are also projected to be present in this stream. The processes being investigated for managing this stream includes evaporation and radionuclide removal via precipitation and adsorption. During evaporation, it is of interest to investigate the formation of insoluble solids to avoid scaling and plugging of equipment. Key parameters for radionuclide removal include identifying effective precipitation or ion

  8. Environmental Management vitrification activities

    Energy Technology Data Exchange (ETDEWEB)

    Krumrine, P.H. [Waste Policy Institute, Gaithersburg, MD (United States)

    1996-05-01

    Both the Mixed Waste and Landfill Stabilization Focus Areas as part of the Office of Technology Development efforts within the Department of Energy`s (DOE) Environmental Management (EM) Division have been developing various vitrification technologies as a treatment approach for the large quantities of transuranic (TRU), TRU mixed and Mixed Low Level Wastes that are stored in either landfills or above ground storage facilities. The technologies being developed include joule heated, plasma torch, plasma arc, induction, microwave, combustion, molten metal, and in situ methods. There are related efforts going into development glass, ceramic, and slag waste form windows of opportunity for the diverse quantities of heterogeneous wastes needing treatment. These studies look at both processing parameters, and long term performance parameters as a function of composition to assure that developed technologies have the right chemistry for success.

  9. Strategy for addressing composition uncertainties in a Hanford high-level waste vitrification plant

    Energy Technology Data Exchange (ETDEWEB)

    Bryan, M.F.; Piepel, G.F.

    1996-03-01

    Various requirements will be imposed on the feed material and glass produced by the high-level waste (HLW) vitrification plant at the Hanford Site. A statistical process/product control system will be used to control the melter feed composition and to check and document product quality. Two general types of uncertainty are important in HLW vitrification process/product control: model uncertainty and composition uncertainty. Model uncertainty is discussed by Hrma, Piepel, et al. (1994). Composition uncertainty includes the uncertainties inherent in estimates of feed composition and other process measurements. Because feed composition is a multivariate quantity, multivariate estimates of composition uncertainty (i.e., covariance matrices) are required. Three components of composition uncertainty will play a role in estimating and checking batch and glass attributes: batch-to-batch variability, within-batch uncertainty, and analytical uncertainty. This document reviews the techniques to be used in estimating and updating composition uncertainties and in combining these composition uncertainties with model uncertainty to yield estimates of (univariate) uncertainties associated with estimates of batch and glass properties.

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

    Energy Technology Data Exchange (ETDEWEB)

    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 mission 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

  11. HLW Flexible jumper materials compatibility evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Skidmore, T. E. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2017-11-13

    H-Tank Farm Engineering tasked SRNL/Materials Science & Technology (MS&T) to evaluate the compatibility of Goodyear Viper® chemical transfer hose with HLW solutions. The hose is proposed as a flexible Safety Class jumper for up to six months service. SRNL/MS&T performed various tests to evaluate the effects of radiation, high pH chemistry and elevated temperature on the hose, particularly the inner liner. Test results suggest an upper dose limit of 50 Mrad for the hose. Room temperature burst pressure values at 50 Mrad are estimated at 600- 800 psi, providing a safety factor of 4.0-5.3X over the anticipated operating pressure of 150 psi and a safety factor of 3.0-4.0X over the working pressure of the hose (200 psi), independent of temperature effects. Radiation effects are minimal at doses less than 10 Mrad. Doses greater than 50 Mrad may be allowed, depending on operating conditions and required safety factors, but cannot be recommended at this time. At 250 Mrad, burst pressure values are reduced to the hose working pressure. At 300 Mrad, burst pressures are below 150 psi. At a bounding continuous dose rate of 57,870 rad/hr, the 50 Mrad dose limit is reached within 1.2 months. Actual dose rates may be lower, particularly during non-transfer periods. Refined dose calculations are therefore recommended to justify longer service. This report details the tests performed and interpretation of the results. Recommendations for shelf-life/storage, component quality verification, and post-service examination are provided.

  12. Selecting a plutonium vitrification process

    Energy Technology Data Exchange (ETDEWEB)

    Jouan, A. [Centre d`Etudes de la Vallee du Rhone, Bagnols sur Ceze (France)

    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 of 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.

  13. Database and Interim Glass Property Models for Hanford HLW Glasses

    Energy Technology Data Exchange (ETDEWEB)

    Hrma, Pavel R.; Piepel, Gregory F.; Vienna, John D.; Cooley, Scott K.; Kim, Dong-Sang; Russell, Renee L.

    2001-07-24

    The purpose of this report is to provide a methodology for an increase in the efficiency and a decrease in the cost of vitrifying high-level waste (HLW) by optimizing HLW glass formulation. This methodology consists in collecting and generating a database of glass properties that determine HLW glass processability and acceptability and relating these properties to glass composition. The report explains how the property-composition models are developed, fitted to data, used for glass formulation optimization, and continuously updated in response to changes in HLW composition estimates and changes in glass processing technology. Further, the report reviews the glass property-composition literature data and presents their preliminary critical evaluation and screening. Finally the report provides interim property-composition models for melt viscosity, for liquidus temperature (with spinel and zircon primary crystalline phases), and for the product consistency test normalized releases of B, Na, and Li. Models were fitted to a subset of the screened database deemed most relevant for the current HLW composition region.

  14. Vitrification of hazardous and radioactive wastes

    Energy Technology Data Exchange (ETDEWEB)

    Bickford, D.F.; Schumacher, R.

    1995-12-31

    Vitrification offers many attractive waste stabilization options. Versatility of waste compositions, as well as the inherent durability of a glass waste form, have made vitrification the treatment of choice for high-level radioactive wastes. Adapting the technology to other hazardous and radioactive waste streams will provide an environmentally acceptable solution to many of the waste challenges that face the public today. This document reviews various types and technologies involved in vitrification.

  15. Chapter 20 Gavi-Automated Vitrification Instrument.

    Science.gov (United States)

    Roy, Tammie K; Brandi, Susanna; Peura, Teija T

    2017-01-01

    Gavi is intended for use in a laboratory or clinic environment for the preparation and vitrification of oocytes, cleavage stage embryos and blastocysts. Gavi is designed to automate the equilibration steps in the vitrification process to minimize the variability that occurs during cryopreservation. This automated process reduces the potential for errors and ensures a standardized, repeatable procedure for vitrification in a controlled, closed-system environment.

  16. Innovative Vitrification for Soil Remediation

    Energy Technology Data Exchange (ETDEWEB)

    Hnat, James G.; Patten, John S.; Jetta, Norman W.

    1996-12-31

    Vortec has successfully completed Phases 1 and 2 of a technology demonstration program for an ''Innovative Fossil Fuel Fired Vitrification Technology for Soil Remediation.'' The principal objective of the program is to demonstrate the ability of a Vortec Cyclone Melting System (CMS) to remediate DOE contaminated soils and other waste forms containing TM RCRA hazardous materials, low levels of radionuclides and TSCA (PCB) containing wastes. The demonstration program will verify the ability of this vitrification process to produce a chemically stable glass final waste form which passes both TCLP and PCT quality control requirements, while meeting all federal and state emission control regulations. The demonstration system is designed to process 36 ton/day of as-received drummed or bulk wastes. The processing capacity equates to approximately 160 barrels/day of waste materials containing 30% moisture at an average weight of 450 lbs./barrel.

  17. Cryopreservation of Endothelial Cells in Various Cryoprotective Agents and Media - Vitrification versus Slow Freezing Methods.

    Directory of Open Access Journals (Sweden)

    Achim von Bomhard

    Full Text Available Vitrification of endothelial cells (MHECT-5 has not previously been compared with controlled slow freezing methods under standardized conditions. To identify the best cryopreservation technique, we evaluated vitrification and standardized controlled-rate -1°C/minute cell freezing in a -80°C freezer and tested four cryoprotective agents (CPA, namely dimethyl sulfoxide (DMSO, ethylene glycol (EG, propylene glycol (PG, and glycerol (GLY, and two media, namely Dulbecco's modified Eagle medium Ham's F-12 (DMEMand K+-modified TiProtec (K+TiP, which is a high-potassium-containing medium. Numbers of viable cells in proliferation were evaluated by the CellTiter 96® AQueous One Solution Cell Proliferation Assay (Promega Corporation, Mannheim, Germany. To detect the exact frozen cell number per cryo vial, DNA content was measured by using Hoechst 33258 dye prior to analysis. Thus, results could be evaluated unconstrained by absolute cell number. Thawed cells were cultured in 25 cm2 cell culture flasks to confluence and examined daily by phase contrast imaging. With regard to cell recovery immediately after thawing, DMSO was the most suitable CPA combined with K+TiP in vitrification (99 ±0.5% and with DMEM in slow freezing (92 ±1.6%. The most viable cells in proliferation after three days of culture were obtained in cells vitrificated by using GLY with K+TiP (308 ±34% and PG with DMEM in slow freezing (280 ±27%.

  18. Vitrification for stability of scrap and residue

    Energy Technology Data Exchange (ETDEWEB)

    Forsberg, C.W. [Oak Ridge National Lab., TN (United States)

    1996-05-01

    A conference breakout discussion was held on the subject of vitrification for stabilization of plutonium scrap and residue. This was one of four such sessions held within the vitrification workshop for participants to discuss specific subjects in further detail. The questions and issues were defined by the participants.

  19. Preliminary hazards analysis -- vitrification process

    Energy Technology Data Exchange (ETDEWEB)

    Coordes, D.; Ruggieri, M.; Russell, J.; TenBrook, W.; Yimbo, P. [Science Applications International Corp., Pleasanton, CA (United States)

    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 construction 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.

  20. Conceptual design requirements for Korean Reference HLW disposal System

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Heui Joo; Choi, Jong Won; Hahn, Pil Son; Lee, Jong Youl; Kim, Kyung Soo; Kim, Sung Ki; Cho, Dong Keun; Lee, Yang

    2005-05-15

    This report outlined the requirements for the conceptual design of KRS(Korean Reference HLW disposal System). The site for the disposal of high-level radioactive wastes has not yet been selected in Korea. Since the KRS should be designed under these circumstances, the necessary requirements which should be determined are studied in the report. The amounts of spent fuels from the nuclear power plants in the long-term national power development plan are projected. With this estimation the disposal rates of CANDU and PWR spent fuels are analyzed and determined. The national and international regulations regarding the disposal of HLW are summarized. The functions of the underground facilities are defined. The representative geological conditions are determined since no site is yet decided in Korea.

  1. Closed system for bovine oocyte vitrification

    Directory of Open Access Journals (Sweden)

    Helena Ševelová

    2012-01-01

    Full Text Available The aim of our study was to develop a vitrification carrier for bovine oocyte cryopreservation. The carrier was to be cheap enough, elementary in its construction and meet contemporary requirements for a safe closed system. In a closed system, a cell is prevented from direct exposure to liquid nitrogen, thus minimizing the risk of cross-contamination. Furthermore, two questions regarding the proper vitrification technique were resolved: if it is necessary to partially denude the oocytes before the vitrification process or whether intact cumulus oocyte complexes should be frozen; and if it is more advantageous to preheat the vitrification solutions to female body temperature (39 °C or to keep them at room temperature. Our results show that it is better to partially denude the oocytes prior to vitrification because cryopreserved intact cumulus oocyte complexes often proved dark, non-homogeneous or fragmented cytoplasm after warming, with many of them having visibly widened perivitelline spaces or fractured zonae pellucidae as a result of extensive damage during vitrification. Consequently, intact cumulus oocyte complexes showed significantly lower numbers of cleavage stage embryos on Day 3 compared to partially denuded oocytes (7.4% and 26%, respectively. On the other hand, the survival rate and following development of fertilized oocytes in preheated vitrification solution were equal to results reached at room temperature conditions. In conclusion, results achieved with the newly developed carrier were comparable to previously published studies and therefore they could be recommended for common use.

  2. Electric arc vitrification of REFIOM

    Energy Technology Data Exchange (ETDEWEB)

    Fautre, R.; Meunier, R. [Electricite de France, Research and Development Div., Les Renardieres, 77 - Moret sur Loing (France)

    1997-07-01

    The REFIOM produced by the neutralization of incineration fumes accounts for 3 to 5 % of incinerated Municipal Solid Waste. Each year, 370,000 tons of REFIOM are produced in France. The product contains pollutants (heavy metals and salts) which must be stabilized before storage in an hazardous waste dump (Class 1 waste dump in France). Since 1992, the Research and Development Division of Electricite de France has been developing an electric arc REFIOM vitrification process which ensures the confinement of polluting elements in a vitrified or crystallized matrix. Reprocessing the elements vaporized during melting allows a complete vitrification of the pollutants and limits the ultimate waste to less than 1 %. This process stabilizes the REFIOM and converts it into inert vitrified granules which can be recycled. Studies are underway to characterize the vitrified product: long term behavior, leaching tests, mechanical and geotechnical tests. The main partners are C.E.A. for long term behavior, SCREG for mechanical tests, C.E.P for leaching tests. The good results obtained confirm the long term durability of the vitrified product. The evolution of the French regulation is required to allow the valorization of the vitrified product for road building purposes. The experience acquired with our pilot furnace allowed us ro define the basic specifications for an industrial pilot. This is a necessary step prior to commercializing the process. (authors)

  3. [Oocyte vitrification in an ART laboratory].

    Science.gov (United States)

    Boyer, P; Montjean, D; Tourame, P; Gervoise-Boyer, M

    2013-09-01

    Oocyte vitrification has been authorized in France after the modification of French bioethics law in July 2011. This evolution will bring a dramatic change in patients' management since, from 2011, infertile couples, oocyte donation and fertility preservation programs will benefit this technique in France. We have introduced oocyte vitrification in our ART laboratory through a validation of the method using Evidence-Based Medicine model: open system Cryotop, Ethylène-glycol 15% and DMSO 15%. Based on our 1-year experience, oocyte vitrification upgrades our daily practice while also minimizing embryo cryoconservation as recommended by the law. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  4. Development of analytical cell support for vitrification at the West Valley Demonstration Project. Topical report

    Energy Technology Data Exchange (ETDEWEB)

    Barber, F.H.; Borek, T.T.; Christopher, J.Z. [and others

    1997-12-01

    Analytical and Process Chemistry (A&PC) support is essential to the high-level waste vitrification campaign at the West Valley Demonstration Project (WVDP). A&PC characterizes the waste, providing information necessary to formulate the recipe for the target radioactive glass product. High-level waste (HLW) samples are prepared and analyzed in the analytical cells (ACs) and Sample Storage Cell (SSC) on the third floor of the main plant. The high levels of radioactivity in the samples require handling them in the shielded cells with remote manipulators. The analytical hot cells and third floor laboratories were refurbished to ensure optimal uninterrupted operation during the vitrification campaign. New and modified instrumentation, tools, sample preparation and analysis techniques, and equipment and training were required for A&PC to support vitrification. Analytical Cell Mockup Units (ACMUs) were designed to facilitate method development, scientist and technician training, and planning for analytical process flow. The ACMUs were fabricated and installed to simulate the analytical cell environment and dimensions. New techniques, equipment, and tools could be evaluated m in the ACMUs without the consequences of generating or handling radioactive waste. Tools were fabricated, handling and disposal of wastes was addressed, and spatial arrangements for equipment were refined. As a result of the work at the ACMUs the remote preparation and analysis methods and the equipment and tools were ready for installation into the ACs and SSC m in July 1995. Before use m in the hot cells, all remote methods had been validated and four to eight technicians were trained on each. Fine tuning of the procedures has been ongoing at the ACs based on input from A&PC technicians. Working at the ACs presents greater challenges than had development at the ACMUs. The ACMU work and further refinements m in the ACs have resulted m in a reduction m in analysis turnaround time (TAT).

  5. Is vitrification standard method of cryopreservation

    Directory of Open Access Journals (Sweden)

    Safak Tavukcuoglu

    2012-09-01

    Full Text Available Cryopreservation of human oocytes and embryos or blastocyts is an important choice in assisted reproduction treatment that leads to an increased cumulative outcome while decreasing other attempts’ costs. It provides an opportunity for patients to have more than one attempt following an ovarian stimulation cycle, thereby decreasing the exposure of patients to exogenous gonadotrophins. Vitrification is a cryopreservation technique that leads to a glass-like solidification. Oocyte, zygote, embryo and blastocyst freezing by vitrification method for cryopreservation have been used for many years beside sperms preservation. Moreover, the use of vitrification technology for ovarian tissue cryopreservation to freeze eggs offers such an elderly women who sometime find more difficulty in conceiving or in maintaining pregnancy till full term because of old age compared to relatively younger women who might get better chances to get a healthy pregnancy. Furthermore, vitrification helps cancer patients who are looking to preserve their fertility later on after completing their treatment.

  6. TESTS WITH HIGH-BISMUTH HLW GLASSES FINAL REPORT VSL-10R1780-1 REV 0 12/13/10

    Energy Technology Data Exchange (ETDEWEB)

    MATLACK KS; KRUGER AA; JOSEPH I; GAN H; KOT WK; CHAUDHURI M; MOHR RK; MCKEOWN DA; BARDAKEI T; GONG W; BUECCHELE AC; PEGG IL

    2011-01-05

    This Final Report describes the testing of glass formulations developed for Hanford High Level Waste (HLW) containing high concentrations of bismuth. In previous work on high-bismuth HLW streams specified by the Office of River Protection (ORP), fully compliant, high waste loading compositions were developed and subjected to melter testing on the DM100 vitrification system. However, during heat treatment according to the Hanford Tank Waste Treatment and Immobilization Plant (WTP) HLW canister centerline cooling (CCC) curves, crucible melts of the high-bismuth glasses were observed to foam. Clearly, such an occurrence during cooling of actual HLW canisters would be highly undesirable. Accordingly, the present work involves larger-scale testing to determine whether this effect occurs under more prototypical conditions, as well as crucible-scale tests to determine the causes and potentially remediate the observed foaming behavior. The work included preparation and characterization of crucible melts designed to determine the underlying causes of the foaming behavior as well as to assess potential mitigation strategies. Testing was also conducted on the DM1200 HLW Pilot melter with a composition previously tested on the DM100 and shown to foam during crucible-scale CCC heat treatment. The DM1200 tests evaluated foaming of glasses over a range of bismuth concentrations poured into temperature-controlled, 55-gallon drums which have a diameter that is close to that of the full-scale WTP HLW canisters. In addition, the DM1200 tests provided the first large-scale melter test data on high-bismuth WTP HLW compositions, including information on processing rates, cold cap behavior and off-gas characteristics, and data from this waste composition on the prototypical DM1200 off-gas treatment system. This work builds on previous work performed at the Vitreous State Laboratory (VSL) for ORP on the same waste composition. The scope of this study was outlined in a Test Plan that was

  7. Advances in vitrification techniques in Japan

    OpenAIRE

    佐々木 憲明; 虎田 真一郎; 五十嵐 寛; 吉岡 正弘

    1986-01-01

    Liquid-fed Joule-heated ceramic melter (LFCM) process for the vitrification of high-level liquid waste (HLLW) is now under development by Power Reactor and Nuclear Fuel Deyelopment Corporation (PNC) in Japan. All developmental works are focused on the vitrification plant which is in the stage of design improvement in succession to the detailed design finished in 1984. The construction of the plant will be started in late 1987. Major development items in process technology in recent years are ...

  8. Melter Throughput Enhancements for High-Iron HLW

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A. A. [Department of Energy, Office of River Protection, Richland, Washington (United States); Gan, Hoa [The Catholic University of America, Washington, DC (United States); Joseph, Innocent [The Catholic University of America, Washington, DC (United States); Pegg, Ian L. [The Catholic University of America, Washington, DC (United States); Matlack, Keith S. [The Catholic University of America, Washington, DC (United States); Chaudhuri, Malabika [The Catholic University of America, Washington, DC (United States); Kot, Wing [The Catholic University of America, Washington, DC (United States)

    2012-12-26

    This report describes work performed to develop and test new glass and feed formulations in order to increase glass melting rates in high waste loading glass formulations for HLW with high concentrations of iron. Testing was designed to identify glass and melter feed formulations that optimize waste loading and waste processing rate while meeting all processing and product quality requirements. The work included preparation and characterization of crucible melts to assess melt rate using a vertical gradient furnace system and to develop new formulations with enhanced melt rate. Testing evaluated the effects of waste loading on glass properties and the maximum waste loading that can be achieved. The results from crucible-scale testing supported subsequent DuraMelter 100 (DM100) tests designed to examine the effects of enhanced glass and feed formulations on waste processing rate and product quality. The DM100 was selected as the platform for these tests due to its extensive previous use in processing rate determination for various HLW streams and glass compositions.

  9. Comparison between Slow Freezing and Vitrification in Terms of Ovarian Tissue Viability in a Bovine Model.

    Science.gov (United States)

    Campos, Ana Luisa Menezes; Guedes, Janaína de Souza; Rodrigues, Jhenifer Kliemchen; Pace, Walter Antônio Prata; Fontoura, Renato Rinco; Caetano, João Pedro Junqueira; Marinho, Ricardo Mello

    2016-07-01

    Objective To assess the viability of bovine ovarian tissue after cryopreservation through either slow freezing or vitrification, and to compare it to that of control tissue by performing morphological analyses. Methods The study included 20 bovine ovarian cortex fragments that were divided into control, vitrification, and slow freezing groups. Each group consisted of four fragments of the same ovary, two fixed without cultivation, and two fixed with cultivation. Tissues were evaluated based on follicular morphology immediately after heating and after 7 days of culture, and compared with the control group. Results A total of 240 fragments were analyzed, generating a sample of 1,344 follicles without cultivation and 552 with cultivation. When the non-cultivated samples were classified as non-atretic follicles, 572 were found in the control group, 289 in the vitrification group, and 373 in the slow freezing group, showing no significant differences. When classified as atretic, 46 follicles were found in the control group, 23 in the vitrification group, and 41 in the slow freezing group, also showing no statistical difference. In the post-culture sample, an evolution of the follicular stages could be observed. This finding was important to support that the follicles considered non-atretic in the non-cultivated group were actually viable in the morphological evaluation. Conclusion With no differences between the protocols, vitrification was shown to be an advanced and alternative method for patients who will undergo treatments that carry the risk of ovarian failure, as the method is less expensive, faster, and more adaptable to laboratory routine. Thieme Publicações Ltda Rio de Janeiro, Brazil.

  10. Managing Viable Knowledge

    NARCIS (Netherlands)

    Achterbergh, J.M.I.M.; Vriens, D.J.

    2002-01-01

    In this paper, Beer's Viable System Model (VSM) is applied to knowledge management. Based on the VSM, domains of knowledge are identified that an organization should possess to maintain its viability. The logic of the VSM is also used to support the diagnosis, design and implementation of the

  11. Modeling of NOx Destruction Options for INEEL Sodium-Bearing Waste Vitrification

    Energy Technology Data Exchange (ETDEWEB)

    Wood, Richard Arthur

    2001-09-01

    Off-gas NOx concentrations in the range of 1-5 mol% are expected as a result of the proposed vitrification of sodium-bearing waste at the Idaho National Engineering and Environmental Laboratory. An existing kinetic model for staged combustion (originally developed for NOx abatement from the calcination process) was updated for application to vitrification offgas. In addition, two new kinetic models were developed to assess the feasibility of using selective non-catalytic reduction (SNCR) or high-temperature alone for NOx abatement. Each of the models was developed using the Chemkin code. Results indicate that SNCR is a viable option, reducing NOx levels to below 1000 ppmv. In addition, SNCR may be capable of simultaneously reducing CO emissions to below 100 ppmv. Results for using high-temperature alone were not as promising, indicating that a minimum NOx concentration of 3950 ppmv is achievable at 3344°F.

  12. Vitrification of copper flotation waste.

    Science.gov (United States)

    Karamanov, Alexander; Aloisi, Mirko; Pelino, Mario

    2007-02-09

    The vitrification of an hazardous iron-rich waste (W), arising from slag flotation of copper production, was studied. Two glasses, containing 30wt% W were melted for 30min at 1400 degrees C. The first batch, labeled WSZ, was obtained by mixing W, blast furnace slag (S) and zeolite tuff (Z), whereas the second, labeled WG, was prepared by mixing W, glass cullet (G), sand and limestone. The glass frits showed high chemical durability, measured by the TCLP test. The crystallization of the glasses was evaluated by DTA. The crystal phases formed were identified by XRD resulting to be pyroxene and wollastonite solid solutions, magnetite and hematite. The morphology of the glass-ceramics was observed by optical and scanning electron microscopy. WSZ composition showed a high rate of bulk crystallization and resulted to be suitable for producing glass-ceramics by a short crystallization heat-treatment. WG composition showed a low crystallization rate and good sinterability; glass-ceramics were obtained by sinter-crystallization of the glass frit.

  13. Innovative vitrification for soil remediation

    Energy Technology Data Exchange (ETDEWEB)

    Jetta, N.W.; Patten, J.S.; Hnat, J.G. [Vortec Corp., Collegeville, PA (United States)

    1995-10-01

    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 I 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 a 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.

  14. Innovative vitrification for soil remediation

    Energy Technology Data Exchange (ETDEWEB)

    Jetta, N.W.; Patten, J.S.; Hnat, J.G. [Vortec Corp., Collegeville, PA (United States)] [and others

    1996-03-01

    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 a 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.

  15. Demonstration of pyrometallurgical processing for metal fuel and HLW

    Energy Technology Data Exchange (ETDEWEB)

    Tadafumi, Koyama; Kensuke, Kinoshita; Takatoshi, Hizikata; Tadashi, Inoue [Central Research Inst. of Electric Power Industry, Komae, Tokyo (Japan); Ougier, M.; Rikard, Malmbeck; Glatz, J.P.; Lothar, Koch [European Commission JRC, Institute fuer Transuranium Elements, Karlsruhe (Germany)

    2001-07-01

    CRIEPI and JRC-ITU have started a joint study on pyrometallurgical processing to demonstrate the capability of this type of process for separating actinide elements from spent fuel and HLW. The equipment dedicated for this experiments has been developed and installed in JRC-ITU. The stainless steel box equipped with tele-manipulators is operated under pure Ar atmosphere, and prepared for later installation in a hot cell. Experiments on pyro-processing of un-irradiated U-Pu-Zr metal alloy fuel by molten salt electrorefining has been carried out. Recovery of U and Pu from this type alloy fuel was first demonstrated with using solid iron cathode and liquid Cd cathode, respectively. (author)

  16. Vitrification preserves murine and human donor cells for generation of tissue-engineered intestine.

    Science.gov (United States)

    Spurrier, Ryan G; Speer, Allison L; Grant, Christa N; Levin, Daniel E; Grikscheit, Tracy C

    2014-08-01

    Short bowel syndrome causes significant morbidity and mortality. Tissue-engineered intestine may serve as a viable replacement. Tissue-engineered small intestine (TESI) has previously been generated in the mouse model from donor cells that were harvested and immediately reimplanted; however, this technique may prove impossible in children who are critically ill, hemodynamically unstable, or septic. We hypothesized that organoid units (OU), multicellular clusters containing epithelium and mesenchyme, could be cryopreserved for delayed production of TESI. OU were isolated from TESI was analyzed by histology and immunofluorescence. After cryopreservation, the viability of murine OU was significantly higher in the vitrification group (93 ± 2%, mean ± standard error of the mean) compared with standard freezing (56 ± 6%) (P TESI was successfully generated from the preserved OU. Hematoxylin and eosin staining demonstrated a mucosa composed of a simple columnar epithelium whereas immunofluorescence staining confirmed the presence of both progenitor and differentiated epithelial cells. Furthermore, beta-2-microglobulin confirmed that the human TESI epithelium originated from human cells. We demonstrated improved multicellular viability after vitrification over conventional cryopreservation techniques and the first successful vitrification of murine and human OU with subsequent TESI generation. Clinical application of this method may allow for delayed autologous implantation of TESI for children in extremis. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Redox Control For Hanford HLW Feeds VSL-12R2530-1, REV 0

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A. A. [Department of Energy, Office of River Protection, Richland, Washington (United States); Matlack, Keith S. [The Catholic University of America, Washington, DC (United States); Pegg, Ian L. [The Catholic University of America, Washington, DC (United States); Kot, Wing K. [The Catholic University of America, Washington, DC (United States); Joseph, Innocent [The Catholic University of America, Washington, DC (United States)

    2012-12-13

    The principal objectives of this work were to investigate the effects of processing simulated Hanford HLW at the estimated maximum concentrations of nitrates and oxalates and to identify strategies to mitigate any processing issues resulting from high concentrations of nitrates and oxalates. This report provides results for a series of tests that were performed on the DM10 melter system with simulated C-106/AY-102 HLW. The tests employed simulated HLW feeds containing variable amounts of nitrates and waste organic compounds corresponding to maximum concentrations proj ected for Hanford HLW streams in order to determine their effects on glass production rate, processing characteristics, glass redox conditions, melt pool foaming, and the tendency to form secondary phases. Such melter tests provide information on key process factors such as feed processing behavior, dynamic effects during processing, processing rates, off-gas amounts and compositions, foaming control, etc., that cannot be reliably obtained from crucible melts.

  18. Vitrification preserves proliferation capacity in human spermatogonia.

    Science.gov (United States)

    Poels, Jonathan; Van Langendonckt, Anne; Many, Marie-Christine; Wese, François-Xavier; Wyns, Christine

    2013-03-01

    Does vitrification of human immature testicular tissue (ITT) have potential benefits for future fertility preservation? Does vitrification of human ITT have potential benefits in an in vivo murine xenotransplantation model? Vitrification is able to maintain proliferation capacity in spermatogonial cells after 6 months of xenografting. Controlled slow-freezing is the procedure currently applied for ITT cryobanking in clinical practice. Vitrification has been proposed as a promising technique for long-term storage of ITT, with a view to preserving spermatogonial stem cells (SSCs) for future fertility restoration in young boys suffering from cancer. After vitrification of ITT, in vitro survival of SSCs was demonstrated, but their functionality was not evaluated. Ten ITT pieces issuing from 10 patients aged 2-12 years were used. Fragments of fresh tissue (serving as controls) and fresh, frozen-thawed and vitrified-warmed testicular pieces xenografted to the scrotum of nude mice for 6 months were compared. Upon graft removal, histological and immunohistochemical analyses were performed to evaluate spermatogonia (SG) (MAGE-A4), intratubular proliferation (Ki67), proliferating SG and Leydig cells (3β-HSD). The entire piece of grafted tissue was assessed in each case. Seminiferous tubules showed good integrity after cryopreservation and xenografting for 6 months in all three groups. Survival of SG and their ability to proliferate was observed by immunohistochemistry in all grafted groups. SG were able to initiate spermatogenesis, but blockage at the pachytene stage was observed. The recovery rate of SG was 3.4 ± 3.8, 4.1 ± 7.3 and 7.3 ± 6.3%, respectively, for fresh, slow-frozen and vitrified-warmed tissue after 6 months of xenografting. The study is limited by the low availability of ITT samples of human origin. The mouse xenotransplantation model needs to be refined to study human spermatogenesis. The findings of the present study have potential implications for

  19. Public Perspectives in the Japanese HLW Disposal Program

    Energy Technology Data Exchange (ETDEWEB)

    Inatsugu, Shigefumi; Takeuchi, Mitsuo; Kato, Toshiaki [Nuclear Waste Management Organization of Japan (NUNIO), Tokyo (Japan)

    2006-09-15

    Following legislation entitled the 'Specified Radioactive Waste Final Disposal Act', the Nuclear Waste Management Organization of Japan (NUMO) was established in October 2000 as the implementing organization for geological disposal of vitrified high-level waste (HLW). Implementation of NUMO's disposal project will be based on three principles: 1) respecting public initiative and opinion, 2) adopting a stepwise approach and 3) ensuring transparency in information disclosure. NUMO has decided to adopt an open solicitation approach to finding volunteer municipalities for Preliminary Investigation Areas (PIAs). The official announcement of the start of the open solicitation program was made in 2002. Although no official applications had been received from volunteer municipalities by the end of 2005, NUMO has been continuing to carry out various activities aimed specifically at public communication and encouraging dialogue about the deep geological disposal project This paper summarizes the results obtained and lessons learned so far and identifies the issues that NUMO must tackle immediately in the areas of communication and dialogue.

  20. Vitrification of Germinal Vesicle Stage Oocytes

    OpenAIRE

    ABE, Yasuyuki; AONO, Nobuya; Hara, Kenshiro; Matsumoto, Hiromichi; BAKHTIYARI, Mehrdad; Sasada, Hiroshi; Sato, Eimei

    2004-01-01

    In order to cryopreserve germinal vesicle (GV) stage oocytes, we first need to develop a novel container for keeping large quantities of GV oocytes, because of collecting them as cumulus oocytes complexes (COCs) that have bigger size and larger volume than oocytes themselves, and second modify a protocol for optimizing vitrification of them. In this mini-review, we describe our recent progress for attaining these objectives. When 65 bovine COCs having GV oocytes could be placed on a sheet of ...

  1. Hanford Waste Vitrification Plant applied technology plan

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, O.L.

    1990-09-01

    This Applied Technology Plan describes the process development, verification testing, equipment adaptation, and waste form qualification technical issues and plans for resolution to support the design, permitting, and operation of the Hanford Waste Vitrification Plant. The scope of this Plan includes work to be performed by the research and development contractor, Pacific Northwest Laboratory, other organizations within Westinghouse Hanford Company, universities and companies with glass technology expertise, and other US Department of Energy sites. All work described in this Plan is funded by the Hanford Waste Vitrification Plant Project and the relationship of this Plan to other waste management documents and issues is provided for background information. Work to performed under this Plan is divided into major areas that establish a reference process, develop an acceptable glass composition envelope, and demonstrate feed processing and glass production for the range of Hanford Waste Vitrification Plant feeds. Included in this work is the evaluation and verification testing of equipment and technology obtained from the Defense Waste Processing Facility, the West Valley Demonstration Project, foreign countries, and the Hanford Site. Development and verification of product and process models and other data needed for waste form qualification documentation are also included in this Plan. 21 refs., 4 figs., 33 tabs.

  2. Successful ongoing pregnancies after vitrification of oocytes.

    Science.gov (United States)

    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.

  3. Vitrification by Ultra-fast Cooling at a Low Concentration of Cryoprotectants in a Quartz Microcapillary: A Study Using Murine Embryonic Stem Cells

    Science.gov (United States)

    He, Xiaoming; Park, Eric Y.H.; Fowler, Alex; Yarmush, Martin L.; Toner, Mehmet

    2009-01-01

    Conventional cryopreservation protocols for slow-freezing or vitrification involve cell injury due to ice formation/cell dehydration or toxicity of high cryoprotectant (CPA) concentrations, respectively. In this study, we developed a novel cryopreservation technique to achieve ultra-fast cooling rates using a quartz microcapillary (QMC). The QMC enabled vitrification of murine embryonic stem (ES) cells using an intracellular cryoprotectant concentration in the range used for slowing freezing (1–2 M). The cryoprotectants used included 2 M 1,2-propanediol (PROH, cell membrane permeable) and 0.5 M extracellular trehalose (cell membrane impermeable). More than 70% of the murine ES cells post-vitrification attached with respect to non-frozen control cells, and the proliferation rates of the two groups were similar. Preservation of undifferentiated properties of the pluripotent murine ES cells post vitrification cryopreservation was verified using three different types of assays: the expression of transcription factor Oct-4, the presentation of the membrane surface glycoprotein SSEA-1, and the elevated expression of the intracellular enzyme alkaline phosphatase. These results indicate that vitrification at a low concentration (2 M) of intracellular cryoprotectants is a viable and effective approach for the cryopreservation of murine embryonic stem cells. PMID:18462712

  4. Vitrification: a solution for the wastes of wastes; La vitrification: ca chauffe pour les ultimes

    Energy Technology Data Exchange (ETDEWEB)

    Guihard, B. [Europlasma, 33 - Saint Medard en Jalles (France)

    1997-07-01

    The incineration of wastes generates other wastes (fly ashes) that concentrate a large amount of polluting substances (heavy metals, salts..). French law requires a stabilization of this kind of wastes before their storage. Today vitrification can be considered as an alternative to the stabilization and storage way, the vitrified products could be seen as an interesting material in the building industry or in road works. A few years ago the municipality of Bordeaux decided to launch a demonstration program and a REFIOM (fly ashes) vitrification unit has been operating since 1997. (A.C.)

  5. Evaluation of high-level waste vitrification feed preparation chemistry for an NCAW simulant, FY 1994: Alternate flowsheets (DRAFT)

    Energy Technology Data Exchange (ETDEWEB)

    Smith, H.D.; Merz, M.D.; Wiemers, K.D.; Smith, G.L.

    1996-02-01

    High-level radioactive waste stored in tanks at the U.S. Department of Energy`s (DOE`s) Hanford Site will be pretreated to concentrate radioactive constituents and fed to the vitrification plant A flowsheet for feed preparation within the vitrification plant (based on the Hanford Waste Vitrification Plant (HWVP) design) called for HCOOH addition during the feed preparation step to adjust rheology and glass redox conditions. However, the potential for generating H{sub 2} and NH{sub 3} during treatment of high-level waste (HLW) with HCOOH was identified at Pacific Northwest Laboratory (PNL). Studies at the University of Georgia, under contract with Savannah River Technology Center (SRTC) and PNL, have verified the catalytic role of noble metals (Pd, Rh, Ru), present in the waste, in the generation of H{sub 2} and NH{sub 3}. Both laboratory-scale and pilot-scale studies at SRTC have documented the H{sub 2} and NH{sub 3} generation phenomenal Because H{sub 2} and NH{sub 3} may create hazardous conditions in the vessel vapor space and offgas system of a vitrification plant, reducing the H{sub 2} generation rate and the NH{sub 3} generation to the lowest possible levels consistent with desired melter feed characteristics is important. The Fiscal Year 1993 and 1994 studies were conducted with simulated (non-radioactive), pre-treated neutralized current acid waste (NCAW). Neutralized current acid waste is a high-level waste originating from the plutonium/uranium extraction (PUREX) plant that has been partially denitrated with sugar, neutralized with NaOH, and is presently stored in double-shell tanks. The non-radioactive simulant used for the present study includes all of the trace components found in the waste, or substitutes a chemically similar element for radioactive or very toxic species. The composition and simulant preparation steps were chosen to best simulate the chemical processing characteristics of the actual waste.

  6. Generalized Test Plan for the Vitrification of Simulated High-Level -Waste Calcine in the Idaho National Laboratory‘s Bench -Scale Cold Crucible Induction Melter

    Energy Technology Data Exchange (ETDEWEB)

    Vince Maio

    2011-08-01

    This Preliminary Idaho National Laboratory (INL) Test Plan outlines the chronological steps required to initially evaluate the validity of vitrifying INL surrogate (cold) High-Level-Waste (HLW) solid particulate calcine in INL's Cold Crucible Induction Melter (CCIM). Its documentation and publication satisfies interim milestone WP-413-INL-01 of the DOE-EM (via the Office of River Protection) sponsored work package, WP 4.1.3, entitled 'Improved Vitrification' The primary goal of the proposed CCIM testing is to initiate efforts to identify an efficient and effective back-up and risk adverse technology for treating the actual HLW calcine stored at the INL. The calcine's treatment must be completed by 2035 as dictated by a State of Idaho Consent Order. A final report on this surrogate/calcine test in the CCIM will be issued in May 2012-pending next fiscal year funding In particular the plan provides; (1) distinct test objectives, (2) a description of the purpose and scope of planned university contracted pre-screening tests required to optimize the CCIM glass/surrogate calcine formulation, (3) a listing of necessary CCIM equipment modifications and corresponding work control document changes necessary to feed a solid particulate to the CCIM, (4) a description of the class of calcine that will be represented by the surrogate, and (5) a tentative tabulation of the anticipated CCIM testing conditions, testing parameters, sampling requirements and analytical tests. Key FY -11 milestones associated with this CCIM testing effort are also provided. The CCIM test run is scheduled to be conducted in February of 2012 and will involve testing with a surrogate HLW calcine representative of only 13% of the 4,000 m3 of 'hot' calcine residing in 6 INL Bin Sets. The remaining classes of calcine will have to be eventually tested in the CCIM if an operational scale CCIM is to be a feasible option for the actual INL HLW calcine. This remaining calcine

  7. Optimization of vitrification protocol for cryopreservation of groundnut

    African Journals Online (AJOL)

    user

    2014-01-08

    Jan 8, 2014 ... Embryonic axes obtained from seeds of four groundnut genotypes were dehydrated in Plant. Vitrification Solution (PVS2) solution ... The seed contain high quality edible oil (44 to 52%), easily digestible protein (26 to ..... in vitro- grown grape (Vitis) by a two-step vitrification protocol. Euphytica 131:299-304.

  8. Vitrification of immature feline oocytes with a commercial kit for bovine embryo vitrification.

    Science.gov (United States)

    Apparicio, M; Ruggeri, E; Luvoni, G C

    2013-04-01

    The aim of this study was to evaluate the suitability of a commercial kit for bovine embryo vitrification for cryopreserving cat oocytes and to evaluate comparatively the effects of its use with slow freezing procedure on cryotolerance in terms of morphology and oocyte resumption of meiosis. Germinal vesicle stage oocytes isolated from cat ovaries were either vitrified (n = 72) using a vitrification kit for bovine embryo or slow frozen (n = 69) by exposing oocyte to ethylene glycol solution before being transferred to a programmable embryo freezer. After thawing and warming, oocytes were cultured for 48 h and then were examined for meiosis resumption using bisbenzimide fluorescent staining (Hoechst 33342). Fresh immature oocytes (n = 92) were used as the control group. The proportion of oocytes recovered in a morphologically normal state after thawing/warming was significantly higher in frozen oocytes (94.5%) than in the vitrified ones (75%, p vitrification compared to 60.9% of those submitted to slow freezing procedure (p bovine embryos retain their capacity to resume meiosis after warming and culture, albeit at lower rates than slow frozen oocytes. Vitrification and slow freezing methods show similar proportions of oocytes with normal morphology after culture, which demonstrate that thawed and warmed oocytes that resist to cryodamage have the same chances to maintain their integrity after 48 h of culture. © 2012 Blackwell Verlag GmbH.

  9. A study on safety assessment methodology for a vitrification plant

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Y. C.; Lee, G. S.; Choi, Y. C.; Kim, G. H. [Yonsei Univ., Seoul (Korea, Republic of)

    2002-03-15

    In this study, the technical and regulatory status of radioactive waste vitrification technologies in foreign and domestic plants is investigated and analyzed, and then significant factors are suggested which must be contained in the final technical guideline or standard for the safety assessment of vitrification plants. Also, the methods to estimate the stability of vitrified waste forms are suggested with property analysis of them. The contents and scope of the study are summarized as follows : survey of the status on radioactive waste vitrification technologies in foreign and domestic plants, survey of the characterization methodology for radioactive waste form, analysis of stability for vitrified waste forms, survey and analysis of technical standards and regulations concerned with them in foreign and domestic plants, suggestion of significant factors for the safety assessment of vitrification plants, submission of regulated technical standard on radioactive waste vitrification plats.

  10. Modelling spent fuel and HLW behaviour in repository conditions

    Energy Technology Data Exchange (ETDEWEB)

    Esparza, A. M.; Esteban, J. A.

    2003-07-01

    The aim of this report is to give the reader an overall insight of the different models, which are used to predict the long-term behaviour of the spent fuels and HLW disposed in a repository. The models must be established on basic data and robust kinetics describing the mechanisms controlling spent fuel alteration/dissolution in a repository. The UO2 matrix, or source term, contains embedded in it the , majority of radionuclides of the spent fuel (some are in the gap cladding). For this reason the SF radionuclides release models play a significant role in the performance assessment of radioactive waste disposal. The differences existing between models published in the literature are due to the conceptual understanding of the processes and the degree of the conservatism used with the parameter values, and the boundary conditions. They mainly differ in their level of simplification and their final objective. Sometimes are focused the show compliance with regulatory requirements, other to support decision making, to increase the level of confidence of public and scientific community, could be empirical, semi-empirical or analytical. The models take into account the experimental results from radionuclides releases and their extrapolation to the very long term. Its necessary a great statistics for have a representative dissolution rate, due at the number of experimental results is not very high and many of them show a great scatter, independently of theirs different compositions by axial and radial variations, due to linear power or local burnup. On the other hand, it is difficult to predict the spent fuel behaviour over the long term, based in short term experiments. In this report is given a little description of the radionuclides distribution in the spent fuel and also in the cladding/pellet gap, grain boundary, cracks and rim zones (the matrix rim zone can be considered with an especial characteristics very different to the rest of the spent fuel), and structural

  11. Cryopreservation: Vitrification and Controlled Rate Cooling.

    Science.gov (United States)

    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

  12. Comparison of radiation hazard of HLW in several spent nuclear fuel reprocessing scenarios

    Directory of Open Access Journals (Sweden)

    Gladilov D.

    2012-10-01

    Full Text Available Radiation hazard of radionuclide has been calculated as a product of Aε where A is an activity of radionuclide and ε is a dose coefficient through ingestion. The values Aε of 18 radionuclide in spent fuel of WWER-440 are calculated. Because the full division of americium and curium from HLW is very complicated a separation americium from curium is considered. It is shown that a separation of americium in a special fraction allows decreasing the radiation hazard of HLW by 97.6% after 1000 years.

  13. ROLE OF MANGANESE REDUCTION/OXIDATION (REDOX) ON FOAMING AND MELT RATE IN HIGH LEVEL WASTE (HLW) MELTERS (U)

    Energy Technology Data Exchange (ETDEWEB)

    Jantzen, C; Michael Stone, M

    2007-03-30

    High-level nuclear waste is being immobilized at the Savannah River Site (SRS) by vitrification into borosilicate glass at the Defense Waste Processing Facility (DWPF). Control of the Reduction/Oxidation (REDOX) equilibrium in the DWPF melter is critical for processing high level liquid wastes. Foaming, cold cap roll-overs, and off-gas surges all have an impact on pouring and melt rate during processing of high-level waste (HLW) glass. All of these phenomena can impact waste throughput and attainment in Joule heated melters such as the DWPF. These phenomena are caused by gas-glass disequilibrium when components in the melter feeds convert to glass and liberate gases such as H{sub 2}O vapor (steam), CO{sub 2}, O{sub 2}, H{sub 2}, NO{sub x}, and/or N{sub 2}. During the feed-to-glass conversion in the DWPF melter, multiple types of reactions occur in the cold cap and in the melt pool that release gaseous products. The various gaseous products can cause foaming at the melt pool surface. Foaming should be avoided as much as possible because an insulative layer of foam on the melt surface retards heat transfer to the cold cap and results in low melt rates. Uncontrolled foaming can also result in a blockage of critical melter or melter off-gas components. Foaming can also increase the potential for melter pressure surges, which would then make it difficult to maintain a constant pressure differential between the DWPF melter and the pour spout. Pressure surges can cause erratic pour streams and possible pluggage of the bellows as well. For these reasons, the DWPF uses a REDOX strategy and controls the melt REDOX between 0.09 {le} Fe{sup 2+}/{summation}Fe {le} 0.33. Controlling the DWPF melter at an equilibrium of Fe{sup +2}/{summation}Fe {le} 0.33 prevents metallic and sulfide rich species from forming nodules that can accumulate on the floor of the melter. Control of foaming, due to deoxygenation of manganic species, is achieved by converting oxidized MnO{sub 2} or Mn

  14. PNL vitrification technology development project high-waste loaded high-level waste glasses for high-temperature melter: Letter report

    Energy Technology Data Exchange (ETDEWEB)

    Kim, D.; Hrma, P.R.

    1996-02-01

    For vitrification of high-level wastes (HLW) at the Hanford Site, a Joule-heated overflow type melter with bottom draining capability and capable of operating at temperatures up to 1500{degrees}C is being developed. The original proposed Hanford Waste Vitrification Plant (HWVP) melter used a 1150{degrees}C processing temperature and was tested using glasses with up to 28 wt% waste oxide loading for NCAW (Neutralized Current Acid Waste). The goal of the high-temperature melter (HTM) is the volume reduction of the final product and increase of the waste processing rate by processing high-waste loaded glasses at higher temperatures. This would dramatically decrease waste disposal and processing costs. The aim of glass development for the HTM is to determine compositions and melting temperatures for processible and acceptable glasses with a high waste loading. Glass property/composition models for viscosity and liquidus temperature developed in the Glass Envelope Definition (GED) study were used. The results of glass formulation and experimental testing are presented for NCAW and DST/SST (Double-Shell Tank/Single-Shell Tank) Blend waste. Although the purpose of this report was to summarize the glass development study with Blend waste only, the results with NCAW were needed because glass development with Blend waste was based on the results from the glass development study with NCAW.

  15. Ultrastructure of human mature oocytes after vitrification

    Directory of Open Access Journals (Sweden)

    M.A. Khalili

    2012-08-01

    Full Text Available Since the introduction of human assisted reproduction, oocyte cryopreservation has been regarded as an attractive option to capitalize the reproductive potential of surplus oocytes and preserve female fertility. However, for two decades the endeavor to store oocytes has been limited by the not yet optimized methodologies, with the consequence of poor clinical outcome or of uncertain reproducibility. Vitrification has been developed as the promising technology of cryopreservation even if slow freezing remains a suitable choice. Nevertheless, the insufficiency of clinical and correlated multidisciplinary data is still stirring controversy on the impact of this technique on oocyte integrity. Morphological studies may actually provide a great insight in this debate. Phase contrast microscopy and other light microscopy techniques, including cytochemistry, provided substantial morphofunctional data on cryopreserved oocyte, but are unable to unraveling fine structural changes. The ultrastructural damage is one of the most adverse events associated with cryopreservation, as an effect of cryo-protectant toxicity, ice crystal formation and osmotic stress. Surprisingly, transmission electron microscopy has attracted only limited attention in the field of cryopreservation. In this review, the subcellular structure of human mature oocytes following vitrification is discussed at the light of most relevant ultrastructural studies.

  16. Vitrification of organics-containing wastes

    Science.gov (United States)

    Bickford, D.F.

    1995-01-01

    A process for stabilizing organics-containing waste materials and recovery metals therefrom, and a waste glass product made according to the process are described. Vitrification of wastes such as organic ion exchange resins, electronic components and the like can be accomplished by mixing at least one transition metal oxide with the wastes, and, if needed, glass formers to compensate for a shortage of silicates or other glass formers in the wastes. The transition metal oxide increases the rate of oxidation of organic materials in the wastes to improve the composition of the glass-forming mixture: at low temperatures, the oxide catalyzes oxidation of a portion of the organics in the waste; at higher temperatures, the oxide dissolves and the resulting oxygen ions oxidize more of the organics; and at vitrification temperatures, the metal ions conduct oxygen into the melt to oxidize the remaining organics. In addition, the transition metal oxide buffers the redox potential of the glass melt so that metals such as Au, Pt, Ag, and Cu separate form the melt in the metallic state and can be recovered. After the metals are recovered, the remainder of the melt is allowed to cool and may subsequently be disposed of. The product has good leaching resistance and can be disposed of in an ordinary landfill, or, alternatively, used as a filler in materials such as concrete, asphalt, brick and tile.

  17. Vitrification facility at the West Valley Demonstration Project

    Energy Technology Data Exchange (ETDEWEB)

    DesCamp, V.A.; McMahon, C.L.

    1996-07-01

    This report is a description of the West Valley Demonstration Project`s vitrification facilities from the establishment of the West Valley, NY site as a federal and state cooperative project to the completion of all activities necessary to begin solidification of radioactive waste into glass by vitrification. Topics discussed in this report include the Project`s background, high-level radioactive waste consolidation, vitrification process and component testing, facilities design and construction, waste/glass recipe development, integrated facility testing, and readiness activities for radioactive waste processing.

  18. Design of microwave vitrification systems for radioactive waste

    Energy Technology Data Exchange (ETDEWEB)

    White, T.L.; Wilson, C.T.; Schaich, C.R. [Oak Ridge National Lab., TN (United States); Bostick, T.L. [Oak Ridge K-25 Site, TN (United States)

    1995-12-31

    Oak Ridge National Laboratory (ORNL) is involved in the research and development of high-power microwave heating systems for the vitrification of Department of Energy (DOE) radioactive sludges. Design criteria for a continuous microwave vitrification system capable of processing a surrogate filtercake sludge representative of a typical waste-water treatment operation are discussed. A prototype 915-MHz, 75-kW microwave vitrification system or ``microwave melter`` is described along with some early experimental results that demonstrate a 4 to 1 volume reduction of a surrogate ORNL filtercake sludge.

  19. Design of microwave vitrification systems for radioactive waste

    Energy Technology Data Exchange (ETDEWEB)

    White, T.L.; Wilson, C.T.; Schaick, C.R. [Oak Ridge National Lab., TN (United States); Bostick, W.D. [Oak Ridge K-25 Site, TN (United States)

    1996-04-01

    Oak Ridge National Laboratory (ORNL) is involved in the research and development of high-power microwave heating systems for the vitrification of DOE radioactive sludges. Design criteria for a continuous microwave vitrification system capable of processing a surrogate filtercake sludge representative of a typical waste-water treatment operation are discussed. A prototype 915 MHz, 75 kW microwave vitrification system or `microwave melter` is described along with some early experimental results that demonstrate a 4 to 1 volume reduction of a surrogate ORNL filtercake sludge.

  20. Disposal of defense spent fuel and HLW from the Idaho Chemical Processing Plant

    Energy Technology Data Exchange (ETDEWEB)

    Ermold, L.F.; Loo, H.H.; Klingler, R.D.; Herzog, J.D.; Knecht, D.A.

    1992-12-01

    Acid high-level radioactive waste (HLW) resulting from fuel reprocessing at the Idaho Chemical Processing Plant (ICPP) for the US Department of Energy (DOE) has been solidified to a calcine since 1963 and stored in stainless steel bins enclosed by concrete vaults. Several different types of unprocessed irradiated DOE-owned fuels are also in storage ate the ICPP. In April, 1992, DOE announced that spent fuel would no longer be reprocessed to recover enriched uranium and called for a shutdown of the reprocessing facilities at the ICPP. A new Spent Fuel and HLW Technology Development program was subsequently initiated to develop technologies for immobilizing ICPP spent fuels and HLW for disposal, in accordance with the Nuclear Waste Policy Act. The Program elements include Systems Analysis, Graphite Fuel Disposal, Other Spent Fuel Disposal, Sodium-Bearing Liquid Waste Processing, Calcine Immobilization, and Metal Recycle/Waste Minimization. This paper presents an overview of the ICPP radioactive wastes and current spent fuels, with an emphasis on the description of HLW and spent fuels requiring repository disposal.

  1. Application of hollow fiber vitrification for cryopreservation of bovine early cleavage stage embryos and porcine morula-blastomeres.

    Science.gov (United States)

    Uchikura, Ayuko; Matsunari, Hitomi; Nakano, Kazuaki; Hatae, Shota; Nagashima, Hiroshi

    2016-04-22

    A novel hollow fiber vitrification (HFV) method was applied to materials that have previously been difficult to cryopreserve, thereby expanding the potential application of this method. The results showed that zona-free porcine morulae and their isolated blastomeres remained viable even after vitrification. The rate of development to blastocysts after vitrification was similar for zona-free and zona-intact morulae (21/23, 91.3% for both). Vitrified blastomeres had a developmental potential equal to that of non-vitrified blastomeres (blastocyst formation rate after reaggregation: 16/17, 94.1% for both). The HFV method was also effective for the cryopreservation of in vitro matured/fertilized bovine embryos at the 2- to 4-cell, 8- to 16-cell and morula stages. The blastocyst formation rates of vitrified embryos (66.1-82.5%) were similar to those of non-vitrified embryos (74.5-82.5%). These results indicate that this novel HFV method is an effective tool for embryo cryopreservation that can enhance current practices in reproductive biology.

  2. Properties of the platinoid fission products during vitrification of high-level radioactive waste

    Science.gov (United States)

    Gong, W.; Lutze, W.; Perez-Cardenas, F.; Matlack, K. S.; Pegg, I. L.

    2006-05-01

    Platinoid fission products present in high-level nuclear wastes present particular challenges to their treatment by vitrification. The platinoid metals Ru, Rh, Pd, and their compounds are sparingly soluble in borosilicate glass melts. During glass melting under oxidizing conditions, the platinoids form small crystals of highly dense solid intermetallic phases and oxides. Under reducing conditions, the platinoids form only intermetallic phases. A fraction of these crystals settles to the bottom of the melting furnace, forming an immobile sludge. The fraction settling reported in the literature is highly variable. In the present work, the fraction settling was found to be >90% under reducing conditions but only 10 to 20% under oxidizing conditions. The thickness of the sludge layer depends on the volume fraction of platinoid crystals in the sludge, which is poorly known (typically ~0.06 under oxidizing conditions). Since the electrical conductivity of the sludge can be >10X that of the melt, in joule-heated melters the presence of such a layer can lead to diversion of the electric current, thereby compromising melter operability. The time to failure by this mechanism is clearly of practical importance. A variety of data are required in order to estimate the time to failure due to this mechanism and such data must be obtained under conditions representative of those in a full-size melting furnace. We have acquired such data using a melting furnace installed in our laboratory. This furnace is a one-third scale prototype of the system to be used for the vitrification of defense HLW at Hanford, WA. In the present work, simulated Hanford HLW material was combined with glass formers to produce a melter feed slurry that was then spiked with the platinoids. Over one thousand chemical and optical analyses were performed on hundreds of samples taken from the feed, various locations inside the furnace, the glass melt during pouring, the solid glass, and various locations along

  3. Vitrification of mouse MII oocytes: Developmental competency using paclitaxel

    Directory of Open Access Journals (Sweden)

    Farzaneh Fesahat

    2016-12-01

    Conclusion: A high concentration of paclitaxel, an anticancer drug, interrupted the mouse oocyte competency when supplemented to vitrification media. Consequently, the optimal concentration of this cytoskeleton stabilizer may improve the post-thawed developmental abilities of oocytes.

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

    Science.gov (United States)

    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.

  5. Defense waste vitrification studies during FY 1980

    Energy Technology Data Exchange (ETDEWEB)

    Bjorklund, W.J.

    1981-08-01

    During FY-1980, Pacific Northwest Laboratory (PNL) tested three vitrification processes on simulated high-level radioactive waste typical of that stored or being produced at US defense facilities. Processes tested included a spray calciner/in-can melter, spray calciner/ceramic melter and direct liquid feeding of a ceramic melter. Tests were made on pilot-scale as well as fullscale equipment. Over 16,000 kg of glass product were produced from 68,000 L of simulated waste. Several compositions were tested, and the glass products were evaluated. Emphasis was placed on determining the processing rates and the ability of the waste to be processed. Off-gas data were collected on several runs. Major conclusions drawn from this test program are divided into processing results, glass-product results, and general information.

  6. Swirl chamber for vitrification of fly ashes

    Directory of Open Access Journals (Sweden)

    Zarzycki Robert

    2017-01-01

    Full Text Available The study presents the concept of a swirl chamber used for vitrification of fly ashes. It assumes the use of coal dust in the process of fly ash melting. The coal dust supplied to the swirl chamber and gasified in the atmosphere of O2, CO2 and H2O allows for obtaining combustible gases composed of CO and H2, which are burnt with the pneumatically supplied fly ash. The above process allows for obtaining a product in the form of a molten slag which does not contain coal grains. The study presents numerical calculations for the process of combustion and gasification of coal dust and opportunities for ensuring adequate parameters in the fly ash melting zone. The combustible gases obtained during the process of gasification can be supplied to the chamber of a pulverized-bed boiler.

  7. Slow-freezing versus vitrification for human ovarian tissue cryopreservation.

    Science.gov (United States)

    Klocke, Silke; Bündgen, Nana; Köster, Frank; Eichenlaub-Ritter, Ursula; Griesinger, Georg

    2015-02-01

    Ovarian tissue can be cryopreserved prior to chemotherapy using either the slow-freezing or the vitrification method; however, the data on the equality of the procedures are still conflicting. In this study, a comparison of the cryo-damage of human ovarian tissue induced by either vitrification or slow-freezing was performed. Ovarian tissue from 23 pre-menopausal patients was cryopreserved with either slow-freezing or vitrification. After thawing/warming, the tissue was histologically and immunohistochemically analyzed and cultured in vitro. During tissue culture the estradiol release was assessed. No significant difference was found in the proportion of high-quality follicles after thawing/warming in the slow-freezing and vitrification group, respectively (72.7 versus 66.7 %, p = 0.733). Estradiol secretion by the ovarian tissue was similar between groups during 18 days in vitro culture (area-under-the-curve 5,411 versus 13,102, p = 0.11). Addition of Sphingosine-1-Phosphate or Activin A to the culture medium did not alter estradiol release in both groups. The proportion of Activated Caspase-3 or 'Proliferating-Cell-Nuclear-Antigen' positive follicles at the end of the culture period was similar between slow-freezing and vitrification. Slow-freezing and vitrification result in similar morphological integrity after cryopreservation, a similar estradiol release in culture, and similar rates of follicular proliferation and apoptosis after culture.

  8. In-situ vitrification of transuranic wastes: systems evaluation and applications assessment

    Energy Technology Data Exchange (ETDEWEB)

    Oma, K.H.; Brown, D.R.; Buelt, J.L.; FitzPatrick, V.F.; Hawley, K.A.; Mellinger, G.B.; Napier, B.A.; Silviera, D.J.; Stein, S.L.; Timmerman, C.L.

    1983-09-01

    Major advantages of in-situ vitrification (ISV) as a means of stabilizing radioactive waste are: long term durability of the waste form; cost effectiveness; safety in terms of minimizing worker and public exposure; and applicability to different kinds of soils and buried wastes. This document describes ISV technology that is available as another viable tool for in place stabilization of waste sites. The following sections correspond to the chapters in the body of this document: description of the ISV process; analysis of the performane of the ISV tests conducted thus far; parameters of the ISV process; cost analysis for the ISV process; analysis of occupational and public exposure; and assessment of waste site applications.

  9. Crystallization In High Level Waste (HLW) Glass Melters: Operational Experience From The Savannah River Site

    Energy Technology Data Exchange (ETDEWEB)

    Fox, K. M. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2014-02-27

    processing strategy for the Hanford Tank Waste Treatment and Immobilization Plant (WTP). The basis of this alternative approach is an empirical model predicting the crystal accumulation in the WTP glass discharge riser and melter bottom as a function of glass composition, time, and temperature. When coupled with an associated operating limit (e.g., the maximum tolerable thickness of an accumulated layer of crystals), this model could then be integrated into the process control algorithms to formulate crystal tolerant high level waste (HLW) glasses targeting higher waste loadings while still meeting process related limits and melter lifetime expectancies. This report provides a review of the scaled melter testing that was completed in support of the Defense Waste Processing Facility (DWPF) melter. Testing with scaled melters provided the data to define the DWPF operating limits to avoid bulk (volume) crystallization in the un-agitated DWPF melter and provided the data to distinguish between spinels generated by K-3 refractory corrosion versus spinels that precipitated from the HLW glass melt pool. This report includes a review of the crystallization observed with the scaled melters and the full scale DWPF melters (DWPF Melter 1 and DWPF Melter 2). Examples of actual DWPF melter attainment with Melter 2 are given. The intent is to provide an overview of lessons learned, including some example data, that can be used to advance the development and implementation of an empirical model and operating limit for crystal accumulation for WTP. Operation of the first and second (current) DWPF melters has demonstrated that the strategy of using a liquidus temperature predictive model combined with a 100 °C offset from the normal melter operating temperature of 1150 °C (i.e., the predicted liquidus temperature (TL) of the glass must be 1050 °C or less) has been successful in preventing any detrimental accumulation of spinel in the DWPF melt pool, and spinel has not been

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

    Science.gov (United States)

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

    2010-01-01

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

  11. Corrosion Behavior of Simulated HLW Glass in the Presence of Magnesium Ion

    Directory of Open Access Journals (Sweden)

    Toshikatsu Maeda

    2011-01-01

    Full Text Available Static leach tests were conducted for simulated HLW glass in MgCl2 solution for up to 92 days to investigate the dissolution mechanism of HLW glass under coastal repository condition. Under the condition that magnesium ion exists in leachate, the dissolution rate of the glass did not decrease with time during leaching, while the rate decreased when the magnesium ion depleted in the leachate. In addition, altered layer including magnesium and silica was observed at the surface of the glass after the leach tests. The present results imply that dissolution of the glass is accompanied with formation of magnesium silicate consuming silica, a glass network former. As a consequence, the glass dissolved with an initial high dissolution rate.

  12. HLW Melter Control Strategy Without Visual Feedback VSL-12R2500-1 Rev 0

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A A. [Department of Energy, Office of River Protection, Richland, Washington (United States); Joseph, Innocent [The Catholic University of America, Washington, DC (United States); Matlack, Keith S. [The Catholic University of America, Washington, DC (United States); Callow, Richard A. [The Catholic University of America, Washington, DC (United States); Abramowitz, Howard [The Catholic University of America, Washington, DC (United States); Pegg, Ian L. [The Catholic University of America, Washington, DC (United States); Brandys, Marek [The Catholic University of America, Washington, DC (United States); Kot, Wing K. [The Catholic University of America, Washington, DC (United States)

    2012-11-13

    Plans for the treatment of high level waste (HL W) at the Hanford Tank Waste Treatment and Immobilization Plant (WTP) are based upon the inventory of the tank wastes, the anticipated performance of the pretreatment processes, and current understanding of the capability of the borosilicate glass waste form [I]. The WTP HLW melter design, unlike earlier DOE melter designs, incorporates an active glass bubbler system. The bubblers create active glass pool convection and thereby improve heat and mass transfer and increase glass melting rates. The WTP HLW melter has a glass surface area of 3.75 m{sup 2} and depth of ~ 1.1 m. The two melters in the HLW facility together are designed to produce up to 7.5 MT of glass per day at 100% availability. Further increases in HL W waste processing rates can potentially be achieved by increasing the melter operating temperature above 1150°C and by increasing the waste loading in the glass product. Increasing the waste loading also has the added benefit of decreasing the number of canisters for storage.

  13. DOE-Managed HLW and SNF Research: FY15 EBS and Thermal Analysis Work Package Status.

    Energy Technology Data Exchange (ETDEWEB)

    Matteo, Edward N. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Hadgu, Teklu [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)

    2015-11-01

    This report examines the technical elements necessary to evaluate EBS concepts and perform thermal analysis of DOE-Managed SNF and HLW in the disposal settings of primary interest – argillite, crystalline, salt, and deep borehole. As the disposal design concept is composed of waste inventory, geologic setting, and engineered concept of operation, the engineered barrier system (EBS) falls into the last component of engineered concept of operation. The waste inventory for DOE-Managed HLW and SNF is closely examined, with specific attention to the number of waste packages, the size of waste packages, and the thermal output per package. As expected, the DOE-Managed HLW and SNF inventory has a much smaller volume, and hence smaller number of canisters, as well a lower thermal output, relative to a waste inventory that would include commercial spent nuclear fuel (CSNF). A survey of available data and methods from previous studies of thermal analysis indicates that, in some cases, thermo-hydrologic modeling will be necessary to appropriately address the problem. This report also outlines scope for FY16 work -- a key challenge identified is developing a methodology to effectively and efficiently evaluate EBS performance in each disposal setting on the basis of thermal analyses results.

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

    Science.gov (United States)

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

    2005-09-01

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

  15. MAVIS: An integrated system for live microscopy and vitrification

    Energy Technology Data Exchange (ETDEWEB)

    Koning, Roman I., E-mail: r.i.koning@lumc.nl [Department of Molecular Cell Biology, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Faas, Frank G. [Department of Molecular Cell Biology, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Boonekamp, Michael; Visser, Bram de; Janse, Jan [Department of Instrumental Development, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Wiegant, Joop C. [Department of Molecular Cell Biology, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Breij, Anna de [Department of Infectious Diseases, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Willemse, Joost [Department of Molecular Cell Biology, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Nibbering, Peter H. [Department of Infectious Diseases, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands); Tanke, Hans J.; Koster, Abraham J. [Department of Molecular Cell Biology, Albinusdreef 2, P.O. Box 9600, 2300 RC, Leiden University Medical Center, Leiden (Netherlands)

    2014-08-01

    Cryo-electron microscopy of vitrified biological samples can provide three-dimensional reconstructions of macromolecules and organelles within bacteria and cells at nanometer scale resolution, even in native conditions. Localization of specific structures and imaging of cellular dynamics in cellular cryo-electron microscopy is limited by (i) the use of cryo-fixation to preserve cellular structures, (ii) the restricted availability of electron dense markers to label molecules inside cells and (iii) the inherent low contrast of cryo electron microscopy. These limitations can be mitigated to a large extend by correlative light and electron microscopy, where the sample is imaged by both light and electron microscopy. Here we present a Microscopy and Vitrification Integrated System (MAVIS) that combines a light microscope with a plunger to vitrify thin specimens. MAVIS provides the capability for fluorescence light microscopic imaging of living cells and bacteria that are adhered to an electron microscopy grid and subsequent vitrification within a time frame of seconds. The instrument allows targeting of dynamic biological events in time and space by fluorescence microscopy for subsequent cryo light and electron microscopy. Here we describe the design and performance of the MAVIS, illustrated with biological examples. - Highlights: • We developed new plunger: a Microscopy and Vitrification Integrated System (MAVIS). • The MAVIS is a new tool for integrating of live microscopy and vitrification. • The MAVIS allows fluorescence LM of living cells and vitrification within seconds. • Here we describe the MAVIS design and performance, and show biological examples.

  16. In situ vitrification of radioactive underground tanks

    Energy Technology Data Exchange (ETDEWEB)

    Koegler, S.S.; Gibby, R.D.; Thompson, L.E.

    1991-10-01

    In situ vitrification (ISV) is a treatment process with great potential for remediating underground tanks previously used for storing radioactive and hazardous chemical wastes at US Department of Energy (DOE) sites. Tests at several scales have demonstrated the utility of ISV for these tanks. An engineering-scale test vitrified a 30-cm-diameter buried steel and concrete tank that contained simulated tank sludge. Hazardous components of the tank sludge were immobilized, or removed and captured in the off-gas treatment system, and the tank walls were melted or incorporated into the ISV block. A pilot-scale ISV test vitrified a 1-m simulated underground tank than contained a simulated refractory sludge. The ISV process completely vitrified the tank, its contents, and the soil below the tank to a depth of 2.4 m, producing a uniform glass and crystalline monolith with an estimated mass of 30 tons. A large-scale underground tank test is scheduled for early 1991. 5 refs., 4 figs.

  17. Feasibility Study for Vitrification of Sodium-Bearing Waste

    Energy Technology Data Exchange (ETDEWEB)

    J. J. Quigley; B. D. Raivo; S. O. Bates; S. M. Berry; D. N. Nishioka; P. J. Bunnell

    2000-09-01

    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 under a Settlement Agreement between the Department of Energy and the State of Idaho. One of the requirements of the Settlement Agreement is the complete calcination (i.e., treatment) of all SBW by December 31, 2012. One of the proposed options for treatment of SBW is vitrification. This study will examine the viability of SBW vitrification. This study describes the process and facilities to treat the SBW, from beginning waste input from INTEC Tank Farm to the final waste forms. Schedules and cost estimates for construction and operation of a Vitrification Facility are included. The study includes a facility layout with drawings, process description and flow diagrams, and preliminary equipment requirements and layouts.

  18. Vitrification of early-stage bovine and equine embryos.

    Science.gov (United States)

    Campos-Chillòn, L F; Suh, T K; Barcelo-Fimbres, M; Seidel, G E; Carnevale, E M

    2009-01-15

    The objectives of this study were to: (1) determine an optimal method and stage of development for vitrification of bovine zygotes or early embryos; and (2) use the optimal procedure for bovine embryos to establish equine pregnancies after vitrification and warming of early embryos. Initially, bovine embryos produced by in-vitro fertilization (IVF) were frozen and vitrified in 0.25mL straws with minimal success. A subsequent experiment was done using two vitrification methods and super open pulled straws (OPS) with 1- or 8-cell bovine embryos. In Method 1 (EG-O), embryos were exposed to 1.5M ethylene glycol (EG) for 5min, 7M ethylene glycol and 0.6M galactose for 30s, loaded in an OPS, and plunged into liquid nitrogen. In Method 2 (EG-DMSO), embryos were exposed to 1.1M ethylene glycol and 1.1M dimethyl sulfoxide (DMSO) for 3min, 2.5M ethylene glycol, 2.5M DMSO and 0.5M galactose for 30s, and loaded and plunged as for EG-O. Cryoprotectants were removed after warming in three steps. One- and eight-cell bovine embryos were cultured for 7 and 4.5 d, respectively, after warming, and control embryos were cultured without vitrification. Cleavage rates of 1-cell embryos were similar (P>0.05) for vitrified and control embryos, although the blastocyst rates for EG-O and control embryos were similar and higher (Pvitrification and warming. In summary, a successful method was established for vitrification of early-stage bovine embryos, and this method was used to establish equine pregnancies after vitrification and warming of 2- to 8-cell embryos produced by ICSI.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-12-15

    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 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

  2. Bulk Vitrification Castable Refractory Block Protection Study

    Energy Technology Data Exchange (ETDEWEB)

    Hrma, Pavel R.; Bagaasen, Larry M.; Beck, Andrew E.; Brouns, Thomas M.; Caldwell, Dustin D.; Elliott, Michael L.; Matyas, Josef; Minister, Kevin BC; Schweiger, Michael J.; Strachan, Denis M.; Tinsley, Bronnie P.; Hollenberg, Glenn W.

    2005-05-01

    Bulk vitrification (BV) was selected for a pilot-scale test and demonstration facility for supplemental treatment to accelerate the cleanup of low-activity waste (LAW) at the Hanford U.S. DOE Site. During engineering-scale (ES) tests, a small fraction of radioactive Tc (and Re, its nonradioactive surrogate) were transferred out of the LAW glass feed and molten LAW glass, and deposited on the surface and within the pores of the castable refractory block (CRB). Laboratory experiments were undertaken to understand the mechanisms of the transport Tc/Re into the CRB during vitrification and to evaluate various means of CRB protection against the deposition of leachable Tc/Re. The tests used Re as a chemical surrogate for Tc. The tests with the baseline CRB showed that the molten LAW penetrates into CRB pores before it converts to glass, leaving deposits of sulfates and chlorides when the nitrate components decompose. Na2O from the LAW reacts with the CRB to create a durable glass phase that may contain Tc/Re. Limited data from a single CRB sample taken from an ES experiment indicate that, while a fraction of Tc/Re is present in the CRB in a readily leachable form, most of the Tc/Re deposited in the refractory is retained in the form of a durable glass phase. In addition, the molten salts from the LAW, mainly sulfates, chlorides, and nitrates, begin to evaporate from BV feeds at temperatures below 800 C and condense on solid surfaces at temperatures below 530 C. Three approaches aimed at reducing or preventing the deposition of soluble Tc/Re within the CRB were proposed: metal lining, sealing the CRB surface with a glaze, and lining the CRB with ceramic tiles. Metal liners were deemed unsuitable because evaluations showed that they can cause unacceptable distortions of the electric field in the BV system. Sodium silicate and a low-alkali borosilicate glaze were selected for testing. The glazes slowed down molten salt condensate penetration, but did little to reduce the

  3. Hanford Waste Vitrification Plant technical manual

    Energy Technology Data Exchange (ETDEWEB)

    Larson, D.E. [ed.; Watrous, R.A.; Kruger, O.L. [and others

    1996-03-01

    A key element of the Hanford waste management strategy is the construction of a new facility, the Hanford Waste Vitrification Plant (HWVP), to vitrify existing and future liquid high-level waste produced by defense activities at the Hanford Site. The HWVP mission is to vitrify pretreated waste in borosilicate glass, cast the glass into stainless steel canisters, and store the canisters at the Hanford Site until they are shipped to a federal geological repository. The HWVP Technical Manual (Manual) documents the technical bases of the current HWVP process and provides a physical description of the related equipment and the plant. The immediate purpose of the document is to provide the technical bases for preparation of project baseline documents that will be used to direct the Title 1 and Title 2 design by the A/E, Fluor. The content of the Manual is organized in the following manner. Chapter 1.0 contains the background and context within which the HWVP was designed. Chapter 2.0 describes the site, plant, equipment and supporting services and provides the context for application of the process information in the Manual. Chapter 3.0 provides plant feed and product requirements, which are primary process bases for plant operation. Chapter 4.0 summarizes the technology for each plant process. Chapter 5.0 describes the engineering principles for designing major types of HWVP equipment. Chapter 6.0 describes the general safety aspects of the plant and process to assist in safe and prudent facility operation. Chapter 7.0 includes a description of the waste form qualification program and data. Chapter 8.0 indicates the current status of quality assurance requirements for the Manual. The Appendices provide data that are too extensive to be placed in the main text, such as extensive tables and sets of figures. The Manual is a revision of the 1987 version.

  4. Bovine oocyte vitrification using the Cryotop method: effect of cumulus cells and vitrification protocol on survival and subsequent development.

    Science.gov (United States)

    Zhou, X L; Al Naib, A; Sun, Da-Wen; Sun, D W; Lonergan, P

    2010-08-01

    The ability to successfully cryopreserve mammalian oocytes has numerous practical, economical and ethical benefits, which may positively impact animal breeding programs and assisted conception in humans. However, oocyte survival and development following vitrification remains poor. The aim of the present study was (1) to evaluate the effect of the presence of cumulus cells on the outcome of vitrification of immature (GV) or mature (MII) bovine oocytes, (2) to compare empirical and theoretical vitrification protocols, and (3) to assess the effect of adding ice blockers to vitrification media on survival and development competence of bovine oocytes following vitrification using the Cryotop method. In Experiment 1, cumulus-enclosed and partially-denuded GV and MII oocytes were vitrified in 15% EG+15% Me(2)SO+0.5M sucrose in two steps. In Experiment 2, GV oocytes were vitrified either as above or using theoretical modeling based on permeability and osmotic tolerance characteristics in 30% EG+11.4% trehalose in three steps or 40% EG+11.4% trehalose in four steps. In Experiment 3, GV oocytes were vitrified in media supplemented or not with 1 of 2 ice blockers (21st Century Medicine, Fontana, CA) 1% X-1000, 1% Z-1000 or both in three steps. In Experiment 1, the survival, cleavage and blastocyst rate of cumulus-enclosed oocytes was significantly higher than those of partially-denuded oocytes when vitrified at the GV stage (93.8% vs. 81.3%, 65.8% vs. 47.3%, 11.3% vs. 4.0%, respectively, P0.05). In conclusion, cumulus-enclosed GV bovine oocytes survived vitrification and subsequently developed at higher rates than MII oocytes using Cryotop method and conventional IVF procedure. Theoretical analysis of permeability characteristics and tolerance limits could not explain the low developmental competence of vitrified oocytes. (c) 2010 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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.

  6. Are We Serious in the US about the Disposal of HLW? - 13561

    Energy Technology Data Exchange (ETDEWEB)

    Neill, Robert H. [New Mexico Environmental Evaluation Group (EEG), 9409 Thornton Ave., N.E., Albuquerque, New Mexico 87109 (United States)

    2013-07-01

    Since all efforts to date to dispose of HLW in the US have been unsuccessful, the following specific actions need to be taken if we are serious about such disposal: - The requirement in the EPA environmental radiation protection standards to predict the behavior of these unwanted residuals for one million years is meaningless. The Standards must be revisited. - Characterize two sites. There are myriad ways a site can be found to be unacceptable. Additionally, the existing HLW inventory requires a second repository. - Congress should specify incentives to states under consideration for a site. Perhaps 5% of total cost would be appropriate. - An independent technical review group should be established in such states to evaluate a proposed repository similar to the New Mexico Environmental Evaluation Group (EEG) for the WIPP Project because the state's interests are not necessarily the same as DOE's. - Acceptance or rejection of a proposed site should be based on technical issues, not social ones. Professionals in this field should present papers identifying the merits of HLW disposal in their own state. The scarcity of such research suggests Not In My Back Yard (NIMBY) syndrome. - Medical diagnostic ionizing radiation exposure to the US public is now 8,000 times greater than radiation exposure from nuclear energy. People accept this believing the benefits outweigh any risks. A major effort needs to focus on both benefits as well as risks of radioactive waste disposal. - DOE needs to announce preferences of host rock formations, incentives for states, and potential consequences should we fail to act. (author)

  7. HIGH ALUMINUM HLW (HIGH LEVEL WASTE ) GLASSES FOR HANFORDS WTP (WASTE TREATMENT PROJECT)

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; BOWAN BW; JOSEPH I; GAN H; KOT WK; MATLACK KS; PEGG IL

    2010-01-04

    This paper presents the results of glass formulation development and melter testing to identify high waste loading glasses to treat high-Al high level waste (HLW) at Hanford. Previous glass formulations developed for this HLW had high waste loadings but their processing rates were lower that desired. The present work was aimed at improving the glass processing rate while maintaining high waste loadings. Glass formulations were designed, prepared at crucible-scale and characterized to determine their properties relevant to processing and product quality. Glass formulations that met these requirements were screened for melt rates using small-scale tests. The small-scale melt rate screening included vertical gradient furnace (VGF) and direct feed consumption (DFC) melter tests. Based on the results of these tests, modified glass formulations were developed and selected for larger scale melter tests to determine their processing rate. Melter tests were conducted on the DuraMelter 100 (DMIOO) with a melt surface area of 0.11 m{sup 2} and the DuraMelter 1200 (DMI200) HLW Pilot Melter with a melt surface area of 1.2 m{sup 2}. The newly developed glass formulations had waste loadings as high as 50 wt%, with corresponding Al{sub 2}O{sub 3} concentration in the glass of 26.63 wt%. The new glass formulations showed glass production rates as high as 1900 kg/(m{sup 2}.day) under nominal melter operating conditions. The demonstrated glass production rates are much higher than the current requirement of 800 kg/(m{sup 2}.day) and anticipated future enhanced Hanford Tank Waste Treatment and Immobilization Plant (WTP) requirement of 1000 kg/(m{sup 2}.day).

  8. Process system evaluation-consolidated letters. Volume 1. Alternatives for the off-gas treatment system for the low-level waste vitrification process

    Energy Technology Data Exchange (ETDEWEB)

    Peurrung, L.M.; Deforest, T.J; Richards, J.R.

    1996-03-01

    This report provides an evaluation of alternatives for treating off-gas from the low-level waste (LLW) melter. The study used expertise obtained from the commercial nonradioactive off-gas treatment industry. It was assumed that contact maintenance is possible, although the subsequent risk to maintenance personnel was qualitatively considered in selecting equipment. Some adaptations to the alternatives described may be required, depending on the extent of contact maintenance that can be achieved. This evaluation identified key issues for the off-gas system design. To provide background information, technology reviews were assembled for various classifications of off-gas treatment equipment, including off-gas cooling, particulate control, acid gas control, mist elimination, NO{sub x} reduction, and SO{sub 2} removal. An order-of-magnitude cost estimate for one of the off-gas systems considered is provided using both the off-gas characteristics associated with the Joule-heated and combustion-fired melters. The key issues identified and a description of the preferred off-gas system options are provided below. Five candidate treatment systems were evaluated. All of the systems are appropriate for the different melting/feed preparations currently being considered. The lowest technical risk is achieved using option 1, which is similar to designs for high-level waste (HLW) vitrification in the Hanford Waste Vitrification Project (HWVP) and the West Valley. Demonstration Project. Option 1 uses a film cooler, submerged bed scrubber (SBS), and high-efficiency mist eliminator (HEME) prior to NO{sub x} reduction and high-efficiency particulate air (HEPA) filtration. However, several advantages were identified for option 2, which uses high-temperature filtration. Based on the evaluation, option 2 was identified as the preferred alternative. The characteristics of this option are described below.

  9. Collaboration, Automation, and Information Management at Hanford High Level Radioactive Waste (HLW) Tank Farms

    Energy Technology Data Exchange (ETDEWEB)

    Aurah, Mirwaise Y.; Roberts, Mark A.

    2013-12-12

    Washington River Protection Solutions (WRPS), operator of High Level Radioactive Waste (HLW) Tank Farms at the Hanford Site, is taking an over 20-year leap in technology, replacing systems that were monitored with clipboards and obsolete computer systems, as well as solving major operations and maintenance hurdles in the area of process automation and information management. While WRPS is fully compliant with procedures and regulations, the current systems are not integrated and do not share data efficiently, hampering how information is obtained and managed.

  10. Evaluation of the Cryotech Vitrification Kit for bovine embryos.

    Science.gov (United States)

    Gutnisky, C; Alvarez, G M; Cetica, P D; Dalvit, G C

    2013-12-01

    The purpose of this work was to assess commercially available Cryotech Vitrification Kit, in terms of survival, in vitro development and pregnancy rate for bovine embryos. Cumulus-oocyte complexes (COCs) were recovered from ovaries obtained from slaughtered cows and then matured in vitro for 22 h. COCs were fertilized by sex-sorted sperm in IVF-mSOF and cultured in IVC-mSOF for 7 days to the blastocyst stage. Blastocysts were vitrified with the Cryotech Vitrification Kit(®) and then either warmed to check viability or transferred to synchronized heifers. We observed 100% survival of the in vitro produced blastocysts and obtained the same pregnancy rate (46.8%) as that obtained using fresh in vitro produced blastocysts. We thus conclude that the Cryotech vitrification method is a valid alternative to other vitrification or slow-cooling methods in the bovine species and that it is ready for livestock production. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Bond-controlled configurational entropy reduction in chemical vitrification.

    Science.gov (United States)

    Corezzi, Silvia; Fioretto, Daniele; Rolla, Pierangelo

    2002-12-12

    Glass formation is usually viewed in terms of physical vitrification: a liquid in a metastable state is cooled or compressed so as to avoid crystallization. However, glasses may also be formed by chemical vitrification, a process involving 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 are based on chemical vitrification. Despite the differences in the molecular processes involved in chemical and physical vitrification, surprising similarities are observed in the slowing down of the dynamics and in the thermodynamical properties of the resulting glasses. Explaining such similarities would improve general understanding of the glass transition and may disclose its universal nature. Here we report dielectric and photon-correlation measurements that reveal the origin of the similarity in the dynamical behaviour of physical and chemical glass formers. We find that the evolution of their configurational restrictions proceeds in a similar manner. In particular, we make a connection between the reduction in configurational entropy and the number of chemical bonds, a quantity that can be controlled in experiments.

  12. Vitrification of caudal fin explants from zebrafish adult specimens.

    Science.gov (United States)

    Cardona-Costa, J; Roig, J; Perez-Camps, M; García-Ximénez, F

    2006-01-01

    No data on vitrification of tissue samples are available in fishes. Three vitrification solutions were compared: V1: 20% ethylene glycol and 20% dimethyl sulphoxide; V2: 25% propylene glycol and 20% dimethyl sulphoxide, and; V3: 20% propylene glycol and 13% methanol, all three prepared in Hanks' buffered salt solution plus 20 percent FBS, following the same one step vitrification procedure developed in mammals. Caudal fin tissue pieces were vitrified into 0.25 ml plastic straws in 30s and stored in liquid nitrogen for 3 days minimum, warmed (10s in nitrogen vapour and 5s in a 25 degree C water bath) and cultured (L-15 plus 20% FBS at 28.5 degree C). At the third day of culture, both attachment and outgrowing rates were recorded. V3 led to the worst results (8% of attachment rate). V1 and V2 allow higher attachment rates (V1: 63% vs V2: 50%. P < 0.05) but not significantly different outgrowing rates (83% to 94%). Vitrification of caudal fin pieces is advantageous in fish biodiversity conservation, particularly in the wild, due to the simplicity of procedure and equipment.

  13. Low-level waste vitrification contact maintenance viability study

    Energy Technology Data Exchange (ETDEWEB)

    Leach, C.E., Westinghouse Hanford

    1996-07-12

    This study investigates the economic viability of contact maintenance in the Low-Level Waste Vitrification Facility, which is part of the Hanford Site Tank Waste Remediation System. This document was prepared by Flour Daniel, Inc., and transmitted to Westinghouse Hanford Company in September 1995.

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

    Science.gov (United States)

    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 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.

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

    Science.gov (United States)

    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.

  16. Vitrification and neomineralisation of bentonitic and kaolinitic clays ...

    African Journals Online (AJOL)

    Resultant fired mineral phases depicted mineral compositions of ceramic bodies, and the study suggested that these clays could be gainfully utilized in the making of ceramic wares, subject to selected beneficiation processes. Keywords: kaolin, bentonite, vitrification, neomineralization, ceramic, firing. Global Journal of ...

  17. Nuclear maturation of immature bovine oocytes after vitrification ...

    African Journals Online (AJOL)

    In the second experiment, effectiveness of both vitrification methods was compared for cryopreservation of immature bovine oocytes. After warming, COCs were cultured in vitro for 24 h. The polar body (PB+) and metaphase-II (MII) stage rates differed significantly among treatment groups. Oocytes vitrified using cryotop ...

  18. Final Report - Glass Formulation Development and Testing for DWPF High AI2O3 HLW Sludges, VSL-10R1670-1, Rev. 0, dated 12/20/10

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Pegg, I. L.; Kot, W. K.; Gan, H.; Matlack, K. S.

    2013-11-13

    The principal objective of the work described in this Final Report is to develop and identify glass frit compositions for a specified DWPF high-aluminum based sludge waste stream that maximizes waste loading while maintaining high production rate for the waste composition provided by ORP/SRS. This was accomplished through a combination of crucible-scale, vertical gradient furnace, and confirmation tests on the DM100 melter system. The DM100-BL unit was selected for these tests. The DM100-BL was used for previous tests on HLW glass compositions that were used to support subsequent tests on the HLW Pilot Melter. It was also used to process compositions with waste loadings limited by aluminum, bismuth, and chromium, to investigate the volatility of cesium and technetium during the vitrification of an HLW AZ-102 composition, to process glass formulations at compositional and property extremes, and to investigate crystal settling on a composition that exhibited one percent crystals at 963{degrees}C (i.e., close to the WTP limit). The same melter was selected for the present tests in order to maintain comparisons between the previously collected data. The tests provide information on melter processing characteristics and off-gas data, including formation of secondary phases and partitioning. Specific objectives for the melter tests are as follows: Determine maximum glass production rates without bubbling for a simulated SRS Sludge Batch 19 (SB19). Demonstrate a feed rate equivalent to 1125 kg/m{sup 2}/day glass production using melt pool bubbling. Process a high waste loading glass composition with the simulated SRS SB19 waste and measure the quality of the glass product. Determine the effect of argon as a bubbling gas on waste processing and the glass product including feed processing rate, glass redox, melter emissions, etc.. Determine differences in feed processing and glass characteristics for SRS SB19 waste simulated by the co-precipitated and direct

  19. Principles of Product Quality Control of German Radioactive Waste Forms from the Reprocessing of Spent Fuel: Vitrification, Compaction and Numerical Simulation - 12529

    Energy Technology Data Exchange (ETDEWEB)

    Tietze-Jaensch, Holger; Schneider, Stephan; Aksyutina, Yuliya; Bosbach, Dirk [Product Quality Control Office for Radioactive Waste (PKS) at the Institute of Energy- and Climate Research, Nuclear Waste Management and Reactor Safety Research, IEK-6, Forschungszentrum Juelich GmbH, 52425 Juelich (Germany); Gauthier, Rene; Eissler, Alexander [WAK Wiederaufarbeitungsanlage Karlsruhe Rueckbau- und Entsorgungs- GmbH, Post Box 1263, 76339 Eggenstein- Leopoldshafen (Germany)

    2012-07-01

    The German product quality control is inter alia responsible for control of two radioactive waste forms of heat generating waste: a) homogeneous vitrified HLW and b) heterogeneous compacted hulls, end-pieces and technological metallic waste. In either case, significantly different metrology is employed at the site of the conditioning plant for the obligatory nuclide inventory declaration. To facilitate an independent evaluation and checking of the accompanying documentation numerical simulations are carried out. The physical and chemical properties of radioactive waste residues are used to assess the data consistency and uncertainty margins, as well as to predict the long-term behavior of the radioactive waste. This is relevant for repository acceptance and safety considerations. Our new numerical approach follows a bottom-up simulation starting from the burn-up behavior of the fuel elements in the reactor core. The output of these burn-up calculations is then coupled with a program that simulates the material separation in the subsequent dissolution and extraction processes normalized to the mass balance. Follow-up simulations of the separated reprocessing lines of a) the vitrification of highly-active liquid and b) the compaction of residual intermediate-active metallic hulls remaining after fuel pellets dissolution, end-pieces and technological waste, allows calculating expectation values for the various repository relevant properties of either waste stream. The principles of the German product quality control of radioactive waste residues from the spent fuel reprocessing have been introduced and explained. Namely, heat generating homogeneous vitrified HLW and heterogeneous compacted metallic MLW have been discussed. The advantages of a complementary numerical property simulation have been made clear and examples of benefits are presented. We have compiled a new program suite to calculate the physical and radio-chemical properties of common nuclear waste

  20. MELT RATE ENHANCEMENT FOR HIGH ALUMINUM HLW (HIGH LEVEL WASTE) GLASS FORMULATION FINAL REPORT 08R1360-1

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; KOT W; PEGG IL; JOSEPH I; BARDAKCI T; GAN H; GONG W; CHAUDHURI M

    2010-01-04

    This report describes the development and testing of new glass formulations for high aluminum waste streams that achieve high waste loadings while maintaining high processing rates. The testing was based on the compositions of Hanford High Level Waste (HLW) with limiting concentrations of aluminum specified by the Office of River Protection (ORP). The testing identified glass formulations that optimize waste loading and waste processing rate while meeting all processing and product quality requirements. The work included preparation and characterization of crucible melts and small scale melt rate screening tests. The results were used to select compositions for subsequent testing in a DuraMelter 100 (DM100) system. These tests were used to determine processing rates for the selected formulations as well as to examine the effects of increased glass processing temperature, and the form of aluminum in the waste simulant. Finally, one of the formulations was selected for large-scale confirmatory testing on the HLW Pilot Melter (DM1200), which is a one third scale prototype of the Hanford Tank Waste Treatment and Immobilization Plant (WTP) HLW melter and off-gas treatment system. This work builds on previous work performed at the Vitreous State Laboratory (VSL) for Department of Energy (DOE) to increase waste loading and processing rates for high-iron HLW waste streams as well as previous tests conducted for ORP on the same high-aluminum waste composition used in the present work and other Hanford HLW compositions. The scope of this study was outlined in a Test Plan that was prepared in response to an ORP-supplied statement of work. It is currently estimated that the number of HLW canisters to be produced in the WTP is about 13,500 (equivalent to 40,500 MT glass). This estimate is based upon the inventory of the tank wastes, the anticipated performance of the sludge treatment processes, and current understanding of the capability of the borosilicate glass waste form

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

    Science.gov (United States)

    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.

  2. Viable Syntax: Rethinking Minimalist Architecture

    Directory of Open Access Journals (Sweden)

    Ken Safir

    2010-03-01

    Full Text Available Hauser et al. (2002 suggest that the human language faculty emerged as a genetic innovation in the form of what is called here a ‘keystone factor’—a single, simple, formal mental capability that, interacting with the pre-existing faculties of hominid ancestors, caused a cascade of effects resulting in the language faculty in modern humans. They take Merge to be the keystone factor, but instead it is posited here that Merge is the pre-existing mechanism of thought made viable by a principle that permits relations interpretable at the interfaces to be mapped onto c-command. The simplified minimalist architecture proposed here respects the keystone factor as closely as possible, but is justified on the basis of linguistic analyses it makes available, including a relativized intervention theory applicable across Case, scope, agreement, selection and linearization, a derivation of the A/A’-distinction from Case theory, and predictions such as why in situ wh-interpretation is island-insensitive, but susceptible to intervention effects.

  3. The Production of Advanced Glass Ceramic HLW Forms using Cold Crucible Induction Melter

    Energy Technology Data Exchange (ETDEWEB)

    Veronica J Rutledge; Vince Maio

    2013-10-01

    Cold Crucible Induction Melters (CCIMs) will favorably change how High-Level radioactive Waste (from nuclear fuel recovery) is treated in the 21st century. Unlike the existing Joule-Heated Melters (JHMs) currently in operation for the glass-based immobilization of High-Level Waste (HLW), CCIMs offer unique material features that will increase melt temperatures, increase throughput, increase mixing, increase loading in the waste form, lower melter foot prints, eliminate melter corrosion and lower costs. These features not only enhance the technology for producing HLW forms, but also provide advantageous attributes to the waste form by allowing more durable alternatives to glass. This paper discusses advantageous features of the CCIM, with emphasis on features that overcome the historical issues with the JHMs presently utilized, as well as the benefits of glass ceramic waste forms over borosilicate glass waste forms. These advantages are then validated based on recent INL testing to demonstrate a first-of-a-kind formulation of a non-radioactive ceramic-based waste form utilizing a CCIM.

  4. Evaluation of the new vacuum infiltration vitrification (viv) cryopreservation technique for native Australian plant shoot tips.

    Science.gov (United States)

    Funnekotter, Bryn; Whiteley, Susan E; Turner, Shane R; Bunn, Eric; Mancera, Ricardo L

    2015-01-01

    The application of a vacuum during the incubation in cryoprotective agents such as PVS2 allows for increased penetration, reducing total incubation times required before vitrification and post-cryopreservation regeneration is achieved. This study compared a conventional droplet-vitrification protocol to the new vacuum infiltration vitrification protocol in four Australian plant species. The new vacuum infiltration vitrification applied an 80 kPa vacuum during incubations in loading solution and PVS2. Infiltration of the cryoprotective agents into shoot tips was determined by differential scanning calorimetry measuring ice formation in the thermographs comparing a range of loading solution and PVS2 incubation times. The application of the vacuum infiltration vitrification technique resulted in a significantly reduced PVS2 incubation time for cryogenic survival and regeneration for all four species, reducing the time needed to adequately protect shoot tips by half to a quarter when compared to a conventional droplet-vitrification technique.

  5. Optimized Method for Bovine Blastocyst Vitrification Using a Simple Hand-Made Cryotip

    OpenAIRE

    Vajiheh Asgari; Mohsen Forouzanfar; Sayed Morteza Hosseini; Mehdi Hajian; Fariba Moulavi; Parvaneh Abedi; Laleh Hosseini; Hossein Sadeghi; Hamid Bahramian; Mohammad Hossein Nasr Esfahani

    2009-01-01

    Objective: This study introduced a simple method for bovine blastocyst vitrification.Materials and Methods: Bovine blastocysts were produced in vitro by means of a wholeco-culture system with vero cells. The blastocysts were randomly divided 1:3 into either vitrification(100 blastocysts) or control (43 blastocysts) groups. For vitrification,expanded - blastocystswere incubated first in equilibration medium for 8 minutes and then in the vitrificationsolution for 1 minute. The blastocysts were ...

  6. Effects of oocyte vitrification on epigenetic status in early bovine embryos.

    Science.gov (United States)

    Chen, Huanhuan; Zhang, Lei; Deng, Tengfei; Zou, Pengda; Wang, Yongsheng; Quan, Fusheng; Zhang, Yong

    2016-08-01

    Oocyte cryopreservation has a great impact on subsequent embryonic development. Currently, several studies have primarily focused on the consequences of vitrification and the development potential of cellular structures. This study determined whether oocyte vitrification caused epigenetic instabilities of bovine embryos. The effects of oocyte vitrification on DNA methylation, histone modifications, and putative imprinted genes' expression in early embryos derived by intracytoplasmic sperm injection were examined. Results showed that oocyte vitrification did not affect zygote cleavage rates (67.0% vs. 73.8% control, P > 0.05) but reduced the blastocyst rate (9.6% vs. 23.0%, P vitrification group during the early cleavage phases. No differences were observed for DNA methylation, H3K9me3, and acH3K9 in the inner cell mass of blastocysts, whereas decreased levels of DNA methylation and acH3K9 (P vitrification. The expression of putative-imprinted genes PEG10, XIST, and KCNQ1O1T was upregulated in blastocysts. These epigenetic abnormalities may be partially explained by altered expression of genes associated with epigenetic regulations. DNA methylation and H3K9 modification suggest that oocyte vitrification may excessively relax the chromosomes of oocytes and early cleavage embryos. In conclusion, these epigenetic indexes could be used as damage markers of oocyte vitrification during early embryonic development, which offers a new insight to assess oocyte vitrification. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Comparison of vitrification and conventional freezing for cryopreservation of caprine embryos.

    Science.gov (United States)

    Araújo-Lemos, Paula F B; Freitas Neto, Leopoldo M; Moura, Marcelo T; Melo, Janaína V; Lima, Paulo F; Oliveira, Marcos A L

    2015-08-01

    The experiment aimed to compare conventional freezing and different vitrification protocols for cryopreservation of caprine embryos at morphological, ultrastructural, and functional levels. Caprine embryos produced in vivo were allocated randomly to three groups: (1) conventional freezing with ethylene glycol (EG); (2) dimethyl sulfoxide + EG (DMSO/EG) vitrification; and (3) dimethylformamide + EG (DMF/EG) vitrification. All groups were scored for cell viability (propidium iodide staining and ultrastructural levels) and re-expansion rate after thawing or warming. Embryos subjected to DMSO/EG vitrification showed higher cell viability (73.33%), compared with DMF/EG vitrification and conventional freezing group embryos (40.00 and 66.66%, respectively). The ultrastructural study revealed that vitrified embryos had greater preservation of cellular structure than embryos from conventional freezing with EG. DMSO/EG vitrification resulted in higher rates of re-expansion in vitro (47.36%) than DMF/EG vitrification (31.58%), and conventional freezing (25.00%). In conclusion, caprine embryos produced in vivo are better cryopreserved after vitrification than conventional freezing, therefore we conclude that DMSO/EG vitrification is the most effective protocol for cryopreservation.

  8. Safeguardability of the vitrification option for disposal of plutonium

    Energy Technology Data Exchange (ETDEWEB)

    Pillay, K.K.S. [Los Alamos National Lab., NM (United States)

    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 and 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.

  9. The role of troublesome components in plutonium vitrification

    Energy Technology Data Exchange (ETDEWEB)

    Li, Hong; Vienna, J.D.; Peeler, D.K.; Hrma, P.; Schweiger, M.J. [Pacific Northwest National Lab., Richland, WA (United States)

    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 issues 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.

  10. Development of a remote bushing for actinide vitrification

    Energy Technology Data Exchange (ETDEWEB)

    Schumacher, R.F.; Ramsey, W.G.; Johnson, F.M. [and others

    1996-12-31

    The Savannah River Site (SRS) and the Savannah River Technology Center (SRTC) are combining their existing experience in handling highly radioactive, special nuclear materials with commercial glass fiberization technology in order to assemble a small vitrification system for radioactive actinide solutions. The vitrification system or {open_quotes}brushing{close_quotes}, is fabricated from platinum-rhodium alloy and is based on early marble remelt fiberization technology. Advantages of this unique system include its relatively small size, reliable operation, geometrical safety (nuclear criticality), and high temperature capability. The bushing design should be capable of vitrifying a number of the actinide nuclear materials, including solutions of americium/curium, neptunium, and possibly plutonium. State of the art, mathematical and oil model studies are being combined with basic engineering evaluations to verify and improve the thermal and mechanical design concepts.

  11. DM100 AND DM1200 MELTER TESTING WITH HIGH WASTE LOADING GLASS FORMULATIONS FOR HANFORD HIGH-ALUMINUM HLW STREAMS

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; KOT WK; PEGG IL; JOSEPH I

    2009-12-30

    This Test Plan describes work to support the development and testing of high waste loading glass formulations that achieve high glass melting rates for Hanford high aluminum high level waste (HLW). In particular, the present testing is designed to evaluate the effect of using low activity waste (LAW) waste streams as a source of sodium in place ofchemical additives, sugar or cellulose as a reductant, boehmite as an aluminum source, and further enhancements to waste processing rate while meeting all processing and product quality requirements. The work will include preparation and characterization of crucible melts in support of subsequent DuraMelter 100 (DM 100) tests designed to examine the effects of enhanced glass formulations, glass processing temperature, incorporation of the LAW waste stream as a sodium source, type of organic reductant, and feed solids content on waste processing rate and product quality. Also included is a confirmatory test on the HLW Pilot Melter (DM1200) with a composition selected from those tested on the DM100. This work builds on previous work performed at the Vitreous State Laboratory (VSL) for Department of Energy's (DOE's) Office of River Protection (ORP) to increase waste loading and processing rates for high-iron HLW waste streams as well as previous tests conducted for ORP on the same waste composition. This Test Plan is prepared in response to an ORP-supplied statement of work. It is currently estimated that the number of HLW canisters to be produced in the Hanford Tank Waste Treatment and Immobilization Plant (WTP) is about 12,500. This estimate is based upon the inventory ofthe tank wastes, the anticipated performance of the sludge treatment processes, and current understanding of the capability of the borosilicate glass waste form. The WTP HLW melter design, unlike earlier DOE melter designs, incorporates an active glass bubbler system. The bubblers create active glass pool convection and thereby improve heat

  12. Independent engineering review of the Hanford Waste Vitrification System

    Energy Technology Data Exchange (ETDEWEB)

    1991-10-01

    The Hanford Waste Vitrification Plant (HWVP) was initiated in June 1987. The HWVP is an essential element of the plan to end present interim storage practices for defense wastes and to provide for permanent disposal. The project start was justified, in part, on efficient technology and design information transfer from the prototype Defense Waste Processing Facility (DWPF). Development of other serial Hanford Waste Vitrification System (HWVS) elements, such as the waste retrieval system for the double-shell tanks (DSTs), and the pretreatment system to reduce the waste volume converted into glass, also was required to accomplish permanent waste disposal. In July 1991, at the time of this review, the HWVP was in the Title 2 design phase. The objective of this technical assessment is to determine whether the status of the technology development and engineering practice is sufficient to provide reasonable assurance that the HWVP and the balance of the HWVS system will operate in an efficient and cost-effective manner. The criteria used to facilitate a judgment of potential successful operation are: vitrification of high-level radioactive waste from specified DSTs on a reasonably continuous basis; and glass produced with physical and chemical properties formally acknowledge as being acceptable for disposal in a repository for high-level radioactive waste. The criteria were proposed specifically for the Independent Engineering Review to focus that assessment effort. They are not represented as the criteria by which the Department will judge the prudence of the Project. 78 refs., 10 figs., 12 tabs.

  13. Development of alternative loading solutions in droplet-vitrification procedures.

    Science.gov (United States)

    Kim, H H; Lee, Y G; Park, S U; Lee, S C; Baek, H J; Cho, E G; Engelmann, F

    2009-01-01

    In plant vitrification protocols, the loading treatment, which involves treating the explants with a moderately concentrated cryoprotectant solution, precedes dehydration of explants with highly concentrated vitrification solutions in order to reduce the toxicity which can be induced by their direct exposure to such highly concentrated solutions. This study aimed at developing alternative loading solutions composed of mixtures of glycerol and sucrose at various concentrations. Differential scanning calorimetry runs of loading solutions and of loaded and dehydrated explants were performed to assay thermal events occurring during cooling and warming. These loading solutions were applied to two model species, viz. garlic and chrysanthemum which were cryopreserved using a droplet-vitrification procedure. The loading treatment proved to be beneficial to both garlic and chrysanthemum and increased recovery of cryopreserved explants. However, response to the loading solutions tested varied between the two model species employed: with garlic, all the loading solutions had a similar effect, whereas survival of chrysanthemum shoot tips was significantly influenced by the composition of the loading solution employed. A loading solution comprising 1.9 M glycerol and 0.5 M sucrose was the most effective. The loading treatment may thus act as an osmotic stress neutralizer and/or induce the physiological adaptation of tissues and cells, including membranes, to both dehydration and freezing.

  14. Improved cryopreservation of chrysanthemum (Chrysanthemum morifolium) using droplet-vitrification.

    Science.gov (United States)

    Lee, Yoon-Geol; Popov, Elena; Cui, Hai-Yan; Kim, Haeng-Hoon; Park, Sang-Un; Bae, Chang-Hyu; Lee, Sheony-Chun; Engelmann, Florent

    2011-01-01

    A droplet-vitrification protocol has been established for cryopreserving Chrysanthemum morifolium cv. Peak using axillary shoot tips and apical shoots of in vitro plants. In the optimized procedure, explants were submitted to a step-wise preculture in liquid sucrose-enriched medium (0.3, 0.5 and 0.7 M for 31,17 and 7 h, respectively). Precultured explants were treated for 40 min with C4 loading solution comprising (w/v) 17.5 percent glycerol + 17.5 percent sucrose, then dehydrated with PVS3 vitrification solution (w/v, 50 percent glycerol + 50 percent sucrose) for 60 min (axillary shoot tips) or 90 min (apical shoots). Explants were cryopreserved by direct immersion in liquid nitrogen in minute drops of PVS3 attached to aluminum foil strips. The optimal age of donor plants was 4-5.5 weeks for apical shoots and 7 weeks for axillary shoot tips, producing post-cryopreservation regeneration percentages of 81.9 percent and 84.9 percernt, respectively. Plants regenerated from cryopreserved samples showed no phenotypical abnormalities and similar profiles of relative DNA content were recorded for control and cryopreserved plants. Our results suggest that the modified droplet-vitrification protocol described in this paper is highly effective and may prove user-friendlier than the cryopreservation protocols already published for chrysanthemum.

  15. Crystallization in high level waste (HLW) glass melters: Savannah River Site operational experience

    Energy Technology Data Exchange (ETDEWEB)

    Fox, Kevin M. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Peeler, David K. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Kruger, Albert A. [USDOE Office of River Protection, Richland, WA (United States)

    2015-06-12

    This paper provides a review of the scaled melter testing that was completed for design input to the Defense Waste Processing Facility (DWPF) melter. Testing with prototype melters provided the data to define the DWPF operating limits to avoid bulk (volume) crystallization in the un-agitated DWPF melter and provided the data to distinguish between spinels generated by refractory corrosion versus spinels that precipitated from the HLW glass melt pool. A review of the crystallization observed with the prototype melters and the full-scale DWPF melters (DWPF Melter 1 and DWPF Melter 2) is included. Examples of actual DWPF melter attainment with Melter 2 are given. The intent is to provide an overview of lessons learned, including some example data, that can be used to advance the development and implementation of an empirical model and operating limit for crystal accumulation for a waste treatment and immobilization plant.

  16. Effect of composition on peraluminous glass properties: An application to HLW containment

    Science.gov (United States)

    Piovesan, V.; Bardez-Giboire, I.; Perret, D.; Montouillout, V.; Pellerin, N.

    2017-01-01

    Part of the Research and Development program concerning high level nuclear waste (HLW) glasses aims to assess new glass formulations able to incorporate a high waste content with enhanced properties in terms of thermal stability, chemical durability, and process ability. This study focuses on peraluminous glasses of the SiO2 - Al2O3 - B2O3 - Na2O - Li2O - CaO - La2O3 system, defined by an excess of aluminum ions Al3+ in comparison with modifier elements such as Na+, Li+ or Ca2+. To understand the effect of composition on physical properties of glasses (viscosity, density, Tg), a Design Of Experiments (DOE) approach was applied to investigate the peraluminous glass domain. The influence of each oxide was quantified to build predictive models for each property. Lanthanum and lithium oxides appear to be the most influential factors on peraluminous glass properties.

  17. Drop Calculations of HLW Canister and Pu Can-in-Canister

    Energy Technology Data Exchange (ETDEWEB)

    Sreten Mastilovic

    2001-07-31

    The objective of this calculation is to determine the structural response of the standard high-level waste (HLW) canister and the canister containing the cans of immobilized plutonium (Pu) (''can-in-canister'' [CIC] throughout this document) subjected to drop DBEs (design basis events) during the handling operation. The evaluated DBE in the former case is 7-m (23-ft) vertical (flat-bottom) drop. In the latter case, two 2-ft (0.61-m) corner (oblique) drops are evaluated in addition to the 7-m vertical drop. These Pu CIC calculations are performed at three different temperatures: room temperature (RT) (20 C ), T = 200 F = 93.3 C , and T = 400 F = 204 C ; in addition to these the calculation characterized by the highest maximum stress intensity is performed at T = 750 F = 399 C as well. The scope of the HLW canister calculation is limited to reporting the calculation results in terms of: stress intensity and effective plastic strain in the canister, directional residual strains at the canister outer surface, and change of canister dimensions. The scope of Pu CIC calculation is limited to reporting the calculation results in terms of stress intensity, and effective plastic strain in the canister. The information provided by the sketches from Reference 26 (Attachments 5.3,5.5,5.8, and 5.9) is that of the potential CIC design considered in this calculation, and all obtained results are valid for this design only. This calculation is associated with the Plutonium Immobilization Project and is performed by the Waste Package Design Section in accordance with Reference 24. It should be noted that the 9-m vertical drop DBE, included in Reference 24, is not included in the objective of this calculation since it did not become a waste acceptance requirement. AP-3.124, ''Calculations'', is used to perform the calculation and develop the document.

  18. DETERMINATION OF HLW GLASS MELT RATE USING X-RAY COMPUTED TOMOGRAPHY

    Energy Technology Data Exchange (ETDEWEB)

    Choi, A.; Miller, D.; Immel, D.

    2011-10-06

    The purpose of the high-level waste (HLW) glass melt rate study is two-fold: (1) to gain a better understanding of the impact of feed chemistry on melt rate through bench-scale testing, and (2) to develop a predictive tool for melt rate in support of the on-going frit development efforts for the Defense Waste Processing Facility (DWPF). In particular, the focus is on predicting relative melt rates, not the absolute melt rates, of various HLW glass formulations solely based on feed chemistry, i.e., the chemistry of both waste and glass-forming frit for DWPF. Critical to the successful melt rate modeling is the accurate determination of the melting rates of various HLW glass formulations. The baseline procedure being used at the Savannah River National Laboratory (SRNL) is to; (1) heat a 4 inch-diameter stainless steel beaker containing a mixture of dried sludge and frit in a furnace for a preset period of time, (2) section the cooled beaker along its diameter, and (3) measure the average glass height across the sectioned face using a ruler. As illustrated in Figure 1-1, the glass height is measured for each of the 16 horizontal segments up to the red lines where relatively large-sized bubbles begin to appear. The linear melt rate (LMR) is determined as the average of all 16 glass height readings divided by the time during which the sample was kept in the furnace. This 'visual' method has proved useful in identifying melting accelerants such as alkalis and sulfate and further ranking the relative melt rates of candidate frits for a given sludge batch. However, one of the inherent technical difficulties of this method is to determine the glass height in the presence of numerous gas bubbles of varying sizes, which is prevalent especially for the higher-waste-loading glasses. That is, how the red lines are drawn in Figure 1-1 can be subjective and, therefore, may influence the resulting melt rates significantly. For example, if the red lines are drawn too low

  19. INTEGRATED DM 1200 MELTER TESTING OF HLW C-106/AY-102 COMPOSITION USING BUBBLERS VSL-03R3800-1 REV 0 9/15/03

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; GONG W; BARDAKCI T; D' ANGELO NA; KOT WK; PEGG IL

    2011-12-29

    This report documents melter and off-gas performance results obtained on the DM1200 HLW Pilot Melter during processing of simulated HLW C-106/AY-102 feed. The principal objectives of the DM1200 melter testing were to determine the achievable glass production rates for simulated HLW C-106/AY-102 feed; determine the effect of bubbling rate on production rate; characterize melter off-gas emissions; characterize the performance of the prototypical off-gas system components as well as their integrated performance; characterize the feed, glass product, and off-gas effluents; and to perform pre- and post test inspections of system components.

  20. Viability of zebrafish (Danio rerio) ovarian follicles after vitrification in a metal container.

    Science.gov (United States)

    Marques, Lis S; Bos-Mikich, Adriana; Godoy, Leandro C; Silva, Laura A; Maschio, Daniel; Zhang, Tiantian; Streit, Danilo P

    2015-12-01

    Cryopreservation of ovarian tissue has been studied for female germline preservation of farm animals and endangered mammalian species. However, there are relatively few reports on cryopreservation of fish ovarian tissue and especially using vitrification approach. Previous studies of our group has shown that the use of a metal container for the cryopreservation of bovine ovarian fragments results in good primordial and primary follicle morphological integrity after vitrification. The aim of this study was to assess the viability and in vitro development of zebrafish follicles after vitrification of fragmented or whole ovaries using the same metal container. In Experiment 1, we tested the follicular viability of five developmental stages following vitrification in four vitrification solutions using fluorescein diacetate and propidium iodide fluorescent probes. These results showed that the highest viability rates were obtained with immature follicles (Stage I) and VS1 (1.5 M methanol + 4.5 M propylene glycol). In Experiment 2, we used VS1 to vitrify different types of ovarian tissue (fragments or whole ovaries) in two different carriers (plastic cryotube or metal container). In this experiment, Stage I follicle survival was assessed following vitrification by vital staining after 24 h in vitro culture. Follicular morphology was analyzed by light microscopy after vitrification. Data showed that the immature follicles morphology was well preserved after cryopreservation. Follicular survival rate was higher (P vitrification devices were compared. Copyright © 2015 Elsevier Inc. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  2. Review of FY2001 Development Work for Vitrification of Sodium Bearing Waste

    Energy Technology Data Exchange (ETDEWEB)

    Barnes, C.M.; Taylor, D.D.

    2002-09-09

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Moore, L. [Rocky Mountain Peace Center, Boulder, CO (United States)

    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.

  4. An improved vitrification protocol for equine immature oocytes, resulting in a first live foal

    NARCIS (Netherlands)

    Ortiz-Escribano, N.; Bogado Pascottini, O.; Woelders, H.; Vandenberghe, L.; Schauwer, De C.; Govaere, J.; Abbeel, Van den E.; Vullers, T.; Ververs, C.; Roels, K.; De Velde, Van M.; Soom, van A.; Smits, K.

    2017-01-01

    Background: The success rate for vitrification of immature equine oocytes is low. Although vitrified-warmed oocytes are able to mature, further embryonic development appears to be compromised. Objectives: The aim of this study was to compare two vitrification protocols, and to examine the effect

  5. Cryopreservation of whole ovaries with vascular pedicles: vitrification or conventional freezing?

    Science.gov (United States)

    Zhang, Jian-Min; Sheng, Yan; Cao, Yong-Zhi; Wang, Hong-Yan; Chen, Zi-Jiang

    2011-05-01

    To compare the efficacy of vitrification and conventional freezing of whole ovaries. Ovaries obtained from 5-year-old female bovines were cryopreserved by conventional freezing, rapid freezing and vitrification. The ovarian cortical strips were cryopreserved by conventional freezing. Follicular viability was assessed using the trypan blue test; the percentage of morphologically normal primordial follicles, hormones concentrations in the culture supernatants, and lactate dehyrogenase levels were measured. The efficacy of cryopreservation of whole ovaries by vitrification was higher than those by conventional freezing and rapid freezing. Conventional freezing of ovarian cortical strips was more effective than cryopreservation of whole ovaries by conventional freezing, rapid freezing, and vitrification. Vitrification seems to be more suitable than conventional freezing for cryopreservation of whole ovaries. However, further studies are required to improve the efficacy of vitrifying whole ovaries.

  6. Pure Quantum Interpretations Are not Viable

    Science.gov (United States)

    Schmelzer, I.

    2011-02-01

    Pure interpretations of quantum theory, which throw away the classical part of the Copenhagen interpretation without adding new structure to its quantum part, are not viable. This is a consequence of a non-uniqueness result for the canonical operators.

  7. Enhanced Sulfate Management in HLW Glass Formulations VSL12R2540-1 REV 0

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A. A. [Department of Energy, Office of River Protection, Richland, Washington (United States); Pegg, Ian L. [The Catholic University of America, Washington, DC (United States); Kot, Wing [The Catholic University of America, Washington, DC (United States); Gan, Hao [The Catholic University of America, Washington, DC (United States); Matlack, Keith S. [The Catholic University of America, Washington, DC (United States)

    2012-11-13

    The Low Activity Waste (LAW) tanks that are scheduled to provide the Hanford Tank Waste Treatment and Immobilization Plant (WTP) with waste feeds contain significant amounts of sulfate. The sulfate content in the LAW feeds is sufficiently high that a separate molten sulfate salt phase may form on top of the glass melt during the vitrification process unless suitable glass formulations are employed and sulfate levels are controlled. Since the formation of the salt phase is undesirable from many perspectives, mitigation approaches had to be developed. Considerable progress has been made and reported by the Vitreous State Laboratory (VSL) in enhancing sulfate incorporation into LAW glass melts and developing strategies to manage and mitigate the risks associated with high-sulfate feeds.

  8. Successful vitrification of bovine blastocysts on paper container.

    Science.gov (United States)

    Kim, Y M; Uhm, S J; Gupta, M K; Yang, J S; Lim, J-G; Das, Z C; Heo, Y T; Chung, H-J; Kong, I-K; Kim, N-H; Lee, H T; Ko, D H

    2012-09-15

    Cryopreservation of bovine embryos can be performed by a variety of methods with variable degree of success. Here, we report a new, easy to perform, simple, inexpensive, and successful method for vitrification of bovine blastocysts. In vitro produced bovine blastocysts were exposed to vitrification solution (5.5 m ethylene glycol, 10% serum and 1% sucrose) in one single step for 20 s, loaded on a paper container prepared from commonly available non-slippery, absorbent writing paper, and then were directly plunged into liquid nitrogen for storage. Vitrified blastocysts were warmed by serial rinsing in 0.5, 0.25 and 0.125 m sucrose solution for 1 min each. Results showed that one step exposure of bovine blastocysts to cryoprotective agents was sufficient to achieve successful cryopreservation. Under these conditions, more than 95% of blastocysts survived the vitrification-warming on paper containers which was significantly higher than those obtained from other containers, such as electron microscope (EM) grid (78.1%), open pulled straw (OPS; 80.2%), cryoloop (76.2%) or plastic straw (73.9%). Embryo transfer of blastocysts vitrified-warmed on paper container resulted in successful conception (19.3%) and full-term live birth of offspring (12.3%) which were lower (P 0.05) to those obtained from blastocysts vitrified-warmed on EM grid (23.3 and 14.2%). Our results, therefore, suggest that paper may be an inexpensive and useful container for the cryopreservation of animal embryos. Copyright © 2012 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2011-11-18

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

  10. Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification

    Science.gov (United States)

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

    2011-01-01

    Cryopreservation of mouse embryos is a technological basis that supports biomedical sciences, because many strains of mice have been produced by genetic modifications and the number is consistently increasing year by year. Its technical development started with slow freezing methods in the 1970s1, then followed by vitrification methods developed in the late 1980s2. Generally, the latter technique is advantageous in its quickness, simplicity, and high survivability of recovered embryos. However, the cryoprotectants contained are highly toxic and may affect subsequent embryo development. Therefore, the technique was not applicable to certain strains of mice, even when the solutions are cooled to 4°C to mitigate the toxic effect during embryo handling. At the RIKEN BioResource Center, more than 5000 mouse strains with different genetic backgrounds and phenotypes are maintained3, and therefore we have optimized a vitrification technique with which we can cryopreserve embryos from many different strains of mice, with the benefits of high embryo survival after vitrifying and thawing (or liquefying, more precisely) at the ambient temperature4. Here, we present a vitrification method for mouse embryos that has been successfully used at our center. The cryopreservation solution contains ethylene glycol instead of DMSO to minimize the toxicity to embryos5. It also contains Ficoll and sucrose for prevention of devitrification and osmotic adjustment, respectively. Embryos can be handled at room temperature and transferred into liquid nitrogen within 5 min. Because the original method was optimized for plastic straws as containers, we have slightly modified the protocol for cryotubes, which are more easily accessible in laboratories and more resistant to physical damages. We also describe the procedure of thawing vitrified embryos in detail because it is a critical step for efficient recovery of live mice. These methodologies would be helpful to researchers and technicians who

  11. INNOVATIVE FOSSIL FUEL FIRED VITRIFICATION TECHNOLOGY FOR SOIL REMEDIATION

    Energy Technology Data Exchange (ETDEWEB)

    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/LLW 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.

  12. INNOVATIVE FOSSIL FUEL FIRED VITRIFICATION TECHNOLOGY FOR SOIL REMEDIATION

    Energy Technology Data Exchange (ETDEWEB)

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

    2001-10-31

    This Final 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/LLW 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 MH/C 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. Because of USEPA policies and regulations that do not require treatment of low level or low-level/PCB contaminated wastes, DOE terminated the project because there is no purported need for this technology.

  13. Final Report - Testing of Optimized Bubbler Configuration for HLW Melter VSL-13R2950-1, Rev. 0, dated 6/12/2013

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Pegg, I. L.; Callow, R. A.; Joseph, I.; Matlack, K. S.; Kot, W. K.

    2013-11-13

    The principal objective of this work was to determine the glass production rate increase and ancillary effects of adding more bubbler outlets to the current WTP HLW melter baseline. This was accomplished through testing on the HLW Pilot Melter (DM1200) at VSL. The DM1200 unit was selected for these tests since it was used previously with several HLW waste streams including the four tank wastes proposed for initial processing at Hanford. This melter system was also used for the development and optimization of the present baseline WTP HLW bubbler configuration for the WTP HLW melter, as well as for MACT testing for both HLW and LAW. Specific objectives of these tests were to: Conduct DM1200 melter testing with the baseline WTP bubbling configuration and as augmented with additional bubblers. Conduct DM1200 melter testing to differentiate the effects of total bubbler air flow and bubbler distribution on glass production rate and cold cap formation. Collect melter operating data including processing rate, temperatures at a variety of locations within the melter plenum space, melt pool temperature, glass melt density, and melter pressure with the baseline WTP bubbling configuration and as augmented with additional bubblers. Collect melter exhaust samples to compare particulate carryover for different bubbler configurations. Analyze all collected data to determine the effects of adding more bubblers to the WTP HLW melter to inform decisions regarding future lid re-designs. The work used a high aluminum HLW stream composition defined by ORP, for which an appropriate simulant and high waste loading glass formulation were developed and have been previously processed on the DM1200.

  14. HLW Feed Delivery AZ101 Batch Transfer to the Private Contractor Transfer and Mixing Process Improvements [Initial Release at Rev 2

    Energy Technology Data Exchange (ETDEWEB)

    DUNCAN, G.P.

    2000-02-28

    The primary purpose of this business case is to provide Operations and Maintenance with a detailed transfer process review for the first High Level Waste (HLW) feed delivery to the Privatization Contractor (PC), AZ-101 batch transfer to PC. The Team was chartered to identify improvements that could be implemented in the field. A significant penalty can be invoked for not providing the quality, quantity, or timely delivery of HLW feed to the PC.

  15. Regional Geologic Evaluations for Disposal of HLW and SNF: The Pierre Shale of the Northern Great Plains

    Energy Technology Data Exchange (ETDEWEB)

    Perry, Frank Vinton [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Kelley, Richard E. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-09-14

    The DOE Spent Fuel and Waste Technology (SWFT) R&D Campaign is supporting research on crystalline rock, shale (argillite) and salt as potential host rocks for disposal of HLW and SNF in a mined geologic repository. The distribution of these three potential repository host rocks is limited to specific regions of the US and to different geologic and hydrologic environments (Perry et al., 2014), many of which may be technically suitable as a site for mined geologic disposal. This report documents a regional geologic evaluation of the Pierre Shale, as an example of evaluating a potentially suitable shale for siting a geologic HLW repository. This report follows a similar report competed in 2016 on a regional evaluation of crystalline rock that focused on the Superior Province of the north-central US (Perry et al., 2016).

  16. Final Report - Sulfate Solubility in RPP-WTP HLW Glasses, VSL-06R6780-1, Rev. 0

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Pegg, I. L.; Feng, A.; Gan, H.; Kot, W. K.

    2013-12-03

    This report describes the results of work and testing specified by Test Specifications 24590-HLW-TSP-RT-01-006 Rev 1, Test Plans VSL-02T7800-1 Rev 1 and Test Exceptions 24590-HLW-TEF-RT-05-00007. The work and any associated testing followed established quality assurance requirements and were conducted as authorized. The descriptions provided in this report are an accurate account of both the conduct of the work and the data collected. Results required by the Test Plans are reported. Also reported are any unusual or anomalous occurrences that are different from the starting hypotheses. The test results and this report have been reviewed and verified.

  17. Hanford Waste Vitrification Plant Project Waste Form Qualification Program Plan

    Energy Technology Data Exchange (ETDEWEB)

    Randklev, E.H.

    1993-06-01

    The US Department of Energy has created a waste acceptance process to help guide the overall program for the disposal of high-level nuclear waste in a federal repository. This Waste Form Qualification Program Plan describes the hierarchy of strategies used by the Hanford Waste Vitrification Plant Project to satisfy the waste form qualification obligations of that waste acceptance process. A description of the functional relationship of the participants contributing to completing this objective is provided. The major activities, products, providers, and associated scheduling for implementing the strategies also are presented.

  18. Vitrification of high-level alumina nuclear waste

    Energy Technology Data Exchange (ETDEWEB)

    Brotzman, J.R.

    1979-01-01

    Borophosphate glass compositions have been developed for the vitrification of a high-alumina calcined defense waste. The effect of substituting SiO/sub 2/, P/sub 2/O/sub 5/ and CuO for B/sub 2/O/sub 3/ on the viscosity and leach resistance was measured. The effect of the alkali to borate ratio and the Li/sub 2/O:Na/sub 2/O ratio on the melt viscosity and leach resistance was also measured.

  19. Vitrification of isolated mice blastomeres using a closed loading device

    Directory of Open Access Journals (Sweden)

    Sharma Rakesh

    2009-02-01

    Full Text Available Abstract Isolated blastomeres obtained by embryo biopsy serve mainly for preimplantation genetic screening. Blastomeres are undifferentiated embryonic cells that include all the embryo genetic information. A lot of developing technologies may benefit by the efficient cryopreservation of blastomeres for future potential use, especially for stem cell culture and differentiation control. We are hereby reporting for the first time the feasibility of preserving individual isolated blastomeres in microvolumes in a closed vitrification system. Using a cryotip and propagation in microvolumes, isolated mice blastomeres were vitrified and warmed with 100% post-warming survival.

  20. Development Of High Waste-Loading HLW Glasses For High Bismuth Phosphate Wastes, VSL-12R2550-1, Rev 0

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A. A. [Department of Energy, Office of River Protection, Richland, Washington (United States); Pegg, Ian L. [The Catholic University of America, Washington, DC (United States); Gan, Hao [The Catholic University of America, Washington, DC (United States); Kot, Wing K. [The Catholic University of America, Washington, DC (United States)

    2012-12-13

    This report presents results from tests with new glass formulations that have been developed for several high Bi-P HLW compositions that are expected to be processed at the WTP that have not been tested previously. WTP HLW feed compositions were reviewed to select waste batches that are high in Bi-P and that are reasonably distinct from the Bi-limited waste that has been tested previously. Three such high Bi-P HLW compositions were selected for this work. The focus of the present work was to determine whether the same type of issues as seen in previous work with high-Bi HLW will be seen in HLW with different concentrations of Bi, P and Cr and also whether similar glass formulation development approaches would be successful in mitigating these issues. New glass compositions were developed for each of the three representative Bi-P HLW wastes and characterized with respect to key processing and product quality properties and, in particular, those relating to crystallization and foaming tendency.

  1. Comparative effects of slow freezing and vitrification on cryosurvival of spermatozoa obtained from west African dwarf goat bucks.

    Science.gov (United States)

    Daramola, J O; Adekunle, E O

    2016-01-01

    Slow freezing and vitrification are used to improve the viability of spermatozoa from various species but comparative effects of these cryoprotocols have never been evaluated for spermatozoa obtained from West African Dwarf (WAD) goat bucks. This study evaluated the comparative effects of slow freezing and vitrification on the viability of spermatozoa of WAD goat bucks. Semen samples collected with the aid of artificial vagina were allocated to slow freezing and vitrification protocols and cryopreserved for 30 days in liquid nitrogen. Consistent higher (Pfreezing compared to vitrification. Abnormal sperm cells and malondialdehyde (MDA) concentrations reduced (Pfreezing compared to vitrification. Higher (Pfreezing compared to vitrification. The findings indicated that spermatozoa obtained from WAD goat bucks were better preserved in slow freezing compared to vitrification.

  2. Final Report - Management of High Sulfur HLW, VSL-13R2920-1, Rev. 0, dated 10/31/2013

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Gan, H.; Pegg, I. L.; Feng, Z.; Gan, H; Joseph, I.; Matlack, K. S.

    2013-11-13

    The present report describes results from a series of small-scale crucible tests to determine the extent of corrosion associated with sulfur containing HLW glasses and to develop a glass composition for a sulfur-rich HLW waste stream, which was then subjected to small-scale melter testing to determine the maximum acceptable sulfate loadings. In the present work, a new glass formulation was developed and tested for a projected Hanford HLW composition with sulfate concentrations high enough to limit waste loading. Testing was then performed on the DM10 melter system at successively higher waste loadings to determine the maximum waste loading without the formation of a separate sulfate salt phase. Small scale corrosion testing was also conducted using the glass developed in the present work, the glass developed in the initial phase of this work [26], and a high iron composition, all at maximum sulfur concentrations determined from melter testing, in order to assess the extent of Inconel 690 and MA758 corrosion at elevated sulfate contents.

  3. Development of geological disposal system; localization of element cost data and cost evaluation on the HLW repository

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Byung Sik; Kim, Kil Jung; Yang, Young Jin; Kim, Sung Chun [KOPEC, Taejeon (Korea)

    2002-03-01

    To estimate Total Life Cycle Cost (TSLCC) for Korea HLW Repository through localization of element cost data, we review and re-organize each basic element cost data for reference repository system, localize various element cost and finally estimate TSLCC considering economic parameters. As results of the study, TSLCC is estimated as 17,167,689 million won, which includes costs for site preparation, surface facilities, underground facilities and management/integration. Since HLW repository Project is an early stage of pre-conceptual design at present, the information of design and project information are not enough to perform cost estimate and cost localization for the Project. However, project cost structure is re-organized based on the local condition and Total System Life Cycle Cost is estimated using the previous cost data gathered from construction experience of the local nuclear power plant. Project results can be used as basic reference data to assume total construction cost for the local HLW repository and should be revised to more reliable cost data with incorporating detail project design information into the cost estimate in a future. 20 refs. (Author)

  4. FINAL REPORT INTEGRATED DM1200 MELTER TESTING USING AZ 102 AND C 106/AY-102 HLW SIMULANTS: HLW SIMULANT VERIFICATION VSL-05R5800-1 REV 0 6/27/05

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; GONG W; BARDAKCI T; D' ANGELO NA; BRANDYS M; KOT WK; PEGG IL

    2011-12-29

    The principal objectives of the DM1200 melter tests were to determine the effects of feed rheology, feed solid content, and bubbler configuration on glass production rate and off-gas system performance while processing the HLW AZ-101 and C-106/AY-102 feed compositions; characterize melter off-gas emissions; characterize the performance of the prototypical off-gas system components, as well as their integrated performance; characterize the feed, glass product, and off-gas effluents; and perform pre- and post test inspections of system components. The specific objectives (including test success criteria) of this testing, along with how each objective was met, are outlined in a table. The data provided in this Final Report address the impacts of HLW melter feed rheology on melter throughput and validation of the simulated HLW melter feeds. The primary purpose of this testing is to further validate/verify the HLW melter simulants that have been used for previous melter testing and to support their continued use in developing melter and off-gas related processing information for the Project. The primary simulant property in question is rheology. Simulants and melter feeds used in all previous melter tests were produced by direct addition of chemicals; these feed tend to be less viscous than rheological the upper-bound feeds made from actual wastes. Data provided here compare melter processing for the melter feed used in all previous DM100 and DM1200 tests (nominal melter feed) with feed adjusted by the feed vendor (NOAH Technologies) to be more viscous, thereby simulating more closely the upperbounding feed produced from actual waste. This report provides results of tests that are described in the Test Plan for this work. The Test Plan is responsive to one of several test objectives covered in the WTP Test Specification for this work; consequently, only part of the scope described in the Test Specification was addressed in this particular Test Plan. For the purpose of

  5. Impact Of Particle Agglomeration On Accumulation Rates In The Glass Discharge Riser Of HLW Melter

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, A. A. [Department of Energy, Office of River Protection, Richland, WA (United States); Rodriguez, C. A. [Pacific Northwest National Laboratory, Richland, WA (United States); Matyas, J. [Pacific Northwest National Laboratory, Richland, WA (United States); Owen, A. T. [Pacific Northwest National Laboratory, Richland, WA (United States); Jansik, D. P. [Pacific Northwest National Laboratory, Richland, WA (United States); Lang, J. B. [Pacific Northwest National Laboratory, Richland, WA (United States)

    2012-11-12

    The major factor limiting waste loading in continuous high-level radioactive waste (HLW) melters is an accumulation of particles in the glass discharge riser during a frequent and periodic idling of more than 20 days. An excessive accumulation can produce robust layers a few centimeters thick, which may clog the riser, preventing molten glass from being poured into canisters. Since the accumulation rate is driven by the size of particles we investigated with x-ray microtomography, scanning electron microscopy, and image analysis the impact of spinel forming components, noble metals, and alumina on the size, concentration, and spatial distribution of particles, and on the accumulation rate. Increased concentrations of Fe and Ni in the baseline glass resulted in the formation of large agglomerates that grew over the time to an average size of ~185+-155 {mu}m, and produced >3 mm thick layer after 120 h at 850 deg C. The noble metals decreased the particle size, and therefore significantly slowed down the accumulation rate. Addition of alumina resulted in the formation of a network of spinel dendrites which prevented accumulation of particles into compact layers.

  6. IMPACT OF PARTICLE AGGLOMERATION ON ACCUMULATION RATES IN THE GLASS DISCHARGE RISER OF HLW MELTER

    Energy Technology Data Exchange (ETDEWEB)

    Matyas, Josef; Jansik, Danielle P.; Owen, Antionette T.; Rodriguez, Carmen P.; Lang, Jesse B.; Kruger, Albert A.

    2013-08-05

    The major factor limiting waste loading in continuous high-level radioactive waste (HLW) melters is an accumulation of particles in the glass discharge riser during a frequent and periodic idling of more than 20 days. An excessive accumulation can produce robust layers a few centimeters thick, which may clog the riser, preventing molten glass from being poured into canisters. Since the accumulation rate is driven by the size of particles we investigated with X-ray microtomography, scanning electron microscopy, and image analysis the impact of spinel forming components, noble metals, and alumina on the size, concentration, and spatial distribution of particles, and on the accumulation rate. Increased concentrations of Fe and Ni in the baseline glass resulted in the formation of large agglomerates that grew over the time to an average size of ~185±155 µm, and produced >3 mm thick layer after 120 h at 850 °C. The noble metals decreased the particle size, and therefore significantly slowed down the accumulation rate. Addition of alumina resulted in the formation of a network of spinel dendrites which prevented accumulation of particles into compact layers.

  7. In vitro evaluation and pregnancy rates after vitrification of in vitro produced bovine embryos.

    Science.gov (United States)

    Martínez, A G; de Matos, D G; Furnus, C C; Brogliatti, G M

    1998-10-01

    The efficacy of different vitrification solutions to cryopreserve in vitro-produced bovine blastocysts was evaluated based on in vitro development of embryos in culture and on in vivo development of embryos transferred into recipients. In the first experiment, 2 vitrification solutions were compared: propylene glycol + glycerol (Pg + Gly) and ethylene glycol + Ficoll + sucrose (EFS). Differences in the overall development and hatching rates in favor of EFS were found (56.4 vs 33.3% and 35.4 vs 13.3%; P vitrification solutions were compared: EFS, modified EFS (EFSm) and ethylene glycol + glycerol (Eg + Gly). The vitrification solutions EFSm and Eg + Gly yield higher hatching rates than did EFS (57.7 vs 59.6 vs 35.7%; P vitrification solutions: EFSm and Eg + Gly. There were no differences between them based on the results obtained after transfer (35.2 vs 43.7%). The vitrification solutions EFSm and Eg + Gly have resulted in good pregnancy rates. These results demonstrated that vitrification can be used successfully in the cryopreservation of in-vitro produced bovine embryos, and it might be considered for use in commercial programs.

  8. Vitrification of human ovarian tissue: effect of different solutions and procedures.

    Science.gov (United States)

    Amorim, Christiani Andrade; David, Anu; Van Langendonckt, Anne; Dolmans, Marie-Madeleine; Donnez, Jacques

    2011-03-01

    To test the effect of different vitrification solutions and procedures on the morphology of human preantral follicles. Pilot study. Gynecology research unit in a university hospital. Ovarian biopsies were obtained from nine women aged 22-35 years. Ovarian tissue fragments were subjected to [1] different vitrification solutions to test their toxicity or [2] different vitrification methods using plastic straws, medium droplets, or solid-surface vitrification before in vitro culture. Number of morphologically normal follicles after toxicity testing or vitrification with the different treatments determined by histologic analysis. In the toxicity tests, only VS3 showed similar results to fresh tissue before and after in vitro culture (fresh controls 1 and 2). In addition, this was the only solution able to completely vitrify. In all vitrification procedures, the percentage of normal follicles was lower than in controls. However, of the three protocols, the droplet method yielded a significantly higher proportion of normal follicles. Our experiments showed VS3 to have no deleterious effect on follicular morphology and to be able to completely vitrify, although vitrification procedures were found to affect human follicles. Nevertheless, the droplet method resulted in a higher percentage of morphologically normal follicles. Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  9. Ash from a pulp mill boiler--characterisation and vitrification.

    Science.gov (United States)

    Ribeiro, Ana S M; Monteiro, Regina C C; Davim, Erika J R; Fernandes, M Helena V

    2010-07-15

    The physical, chemical and mineralogical characterisation of the ash resulting from a pulp mill boiler was performed in order to investigate the valorisation of this waste material through the production of added-value glassy materials. The ash had a particle size distribution in the range 0.06-53 microm, and a high amount of SiO(2) (approximately 82 wt%), which was present as quartz. To favour the vitrification of the ash and to obtain a melt with an adequate viscosity to cast into a mould, different amounts of Na(2)O were added to act as fluxing agent. A batch with 80 wt% waste load melted at 1350 degrees C resulting in a homogeneous transparent green-coloured glass with good workability. The characterisation of the produced glass by differential thermal analysis and dilatometry showed that this glass presents a stable thermal behaviour. Standard leaching tests revealed that the concentration of heavy metals in the leaching solution was lower than those allowed by the Normative. As a conclusion, by vitrification of batch compositions with adequate waste load and additive content it is possible to produce an ash-based glass that may be used in similar applications as a conventional silicate glass inclusively as a building ecomaterial. 2010 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    Adu-Gyamfi, Raphael; Wetten, Andy

    2012-01-01

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

  11. Chemical durability of glasses obtained by vitrification of industrial wastes.

    Science.gov (United States)

    Pisciella, P; Crisucci, S; Karamanov, A; Pelino, M

    2001-01-01

    The vitrification of zinc-hydrometallurgy wastes, electric arc furnace dust (EAFD), drainage mud, and granite mud was shown to immobilize the hazardous components in these wastes. Batch compositions were prepared by mixing the wastes with glass-cullet and sand to force the final glass composition into the glass forming region of the SiO2-Fe2O3-(CaO, MgO) system. The vitrification was carried out in the 1400-1450 degrees C temperature range followed by quenching in water or on stainless steel mold. The United States (US) Environmental Protection Agency (EPA) toxic characterization leaching procedure (TCLP) test was used as a standard method for evaluating the leachability of the elements in the glasses and glass-ceramics samples made with different percentages of wastes. The results for EAFD glasses highlighted that the chemical stability is influenced by the glass structure formed, which, in turn, depends on the Si/O ratio in the glass. The chemical durability of jarosite glasses and glass-ceramics was evaluated by 24 h contact in NaOH, HCl and Na2CO3, at 95 degrees C. Jarosite glass-ceramics containing pyroxene (J40) are more durable than the parent glass in HCl. Jarosite glass-ceramics containing magnetite type spinels (J50) have a durability similar to the parent glass and even lower in HCl because the magnetite is soluble in HCl.

  12. MAVIS: an integrated system for live microscopy and vitrification.

    Science.gov (United States)

    Koning, Roman I; Faas, Frank G; Boonekamp, Michael; de Visser, Bram; Janse, Jan; Wiegant, Joop C; de Breij, Anna; Willemse, Joost; Nibbering, Peter H; Tanke, Hans J; Koster, Abraham J

    2014-08-01

    Cryo-electron microscopy of vitrified biological samples can provide three-dimensional reconstructions of macromolecules and organelles within bacteria and cells at nanometer scale resolution, even in native conditions. Localization of specific structures and imaging of cellular dynamics in cellular cryo-electron microscopy is limited by (i) the use of cryo-fixation to preserve cellular structures, (ii) the restricted availability of electron dense markers to label molecules inside cells and (iii) the inherent low contrast of cryo electron microscopy. These limitations can be mitigated to a large extend by correlative light and electron microscopy, where the sample is imaged by both light and electron microscopy. Here we present a Microscopy and Vitrification Integrated System (MAVIS) that combines a light microscope with a plunger to vitrify thin specimens. MAVIS provides the capability for fluorescence light microscopic imaging of living cells and bacteria that are adhered to an electron microscopy grid and subsequent vitrification within a time frame of seconds. The instrument allows targeting of dynamic biological events in time and space by fluorescence microscopy for subsequent cryo light and electron microscopy. Here we describe the design and performance of the MAVIS, illustrated with biological examples. © 2013 Published by Elsevier B.V.

  13. Vitrification of human germinal vesicle oocytes; before or after in vitro maturation?

    Directory of Open Access Journals (Sweden)

    Evangelia Kasapi

    2017-03-01

    Full Text Available Background The use of immature oocytes derived from stimulated cycles could be of great importance, particularly for urgent fertility preservation cases. The current study aimed to determine whether in vitro maturation (IVM was more successful before or after vitrification of these oocytes. Materials and Methods This prospective study was performed in a private in vitro fertilization (IVF center. We collected 318 germinal vesicle (GV oocytes from 104 stimulated oocyte donation cycles. Oocytes were divided into two groups according to whether vitrification was applied at the GV stage (group 1 or in vitro matured to the metaphase II (MII stage and then vitrified (group 2. In the control group (group 3, oocytes were in vitro matured without vitrification. In all three groups, we assessed survival rate after warming, maturation rate, and MII-spindle/chromosome configurations. The chi-square test was used to compare rates between the three groups. Statistical significance was defined at P<0.05 and we used Bonferroni criterion to assess statistical significance regarding the various pairs of groups. The Statistical Package for the Social Sciences version 17.0 was used to perform statistical analysis. Results There was no significant difference in the survival rate after vitrification and warming of GV (93.5% and MII oocytes (90.8%. A significantly higher maturation rate occurred when IVM was performed before vitrification (82.9% compared to after vitrification (51%. There was no significant difference in the incidence of normal spindle/ chromosome configurations among warmed oocytes matured in vitro before (50.0% or after (41.2% vitrification. However, a higher incidence of normal spindle/chromosome configurations existed in the in vitro matured oocytes which were not subjected to vitrification (fresh oocytes, 77.9%. Conclusion In stimulated cycles, vitrification of in vitro matured MII oocytes rather than GV oocytes seems to be more efficient. This

  14. Using Generic Examples to Make Viable Arguments

    Science.gov (United States)

    Adams, Anne E.; Ely, Rob; Yopp, David

    2017-01-01

    The twenty-first century has seen an increased call to train students to craft mathematical arguments. The third of the Common Core's (CCSS) Standards for Mathematical Practice (SMP 3) (CCSSI 2010) calls for all mathematically proficient students to "construct viable arguments" to support the truth of their ideas and to "critique…

  15. Engineering for Operation of a Future Belgian Deep Geological Repository for ILW and HLW - 12379

    Energy Technology Data Exchange (ETDEWEB)

    Haverkamp, B.; Biurrun, E.; Nieder-Westermann, G.H. [DBE TECHNOLOGY GmbH, Peine (Germany); Van Humbeeck, H. [ONDRAF/NIRAS, Brussels (Belgium); Van Cotthem, Alain [Tractebel Engineering SA, Brussels (Belgium)

    2012-07-01

    In Belgium, an advanced conceptual design is being elaborated for deep geologic disposal of high level waste (HLW) and for low and intermediate level waste (LILW) not amenable for surface disposal. The concept is based on a shielded steel and concrete container for disposal of HLW, i.e., the Super-container. LILW will be disposed of in separately designed concrete caissons. The reference host rock is the Boom Clay, a poorly indurated clay formation in northeastern Belgium. Investigations into the potential host rock are conducted at the HADES underground research laboratory in Mol, Belgium. In 2009 the Belgian Agency for Management of Radioactive Waste and Enriched Fissile Materials (ONDRAF/NIRAS) initiated a four year research project aimed at confirming the fundamental feasibility of building and operating a repository. The goal of the program is to demonstrate at a detailed conceptual level that the proposed geologic disposal system can be safely constructed, operated, and progressively closed. Part of the broader research efforts being conducted includes evaluations optimization of the waste transportation shaft, subsurface transportation system, ventilation system, and evaluation of backfilling and sealing concepts for the repository design. The potential for implementation of a waste retrieval strategy encompassing the first 100 years after emplacement is also considered. In the framework of a four year research program aimed at confirming the fundamental feasibility of building and operating a repository in poorly indurated clay design studies have been underway to optimize the waste transportation shaft, subsurface transportation system, and ventilation system. Additionally backfilling and sealing concepts proposed for the potential repository have been reviewed in conjunction with impacts related to the potential future inclusion of a retrievability requirement in governing regulations. The main engineering challenges in the Belgian repository concept are

  16. Hanford Supplemental Treatment: Literature and Modeling Review of SRS HLW Salt Dissolution and Fractional Crystallization

    Energy Technology Data Exchange (ETDEWEB)

    Choi, A. S.; Flach, G. P.; Martino, C. J.; Zamecnik, J. R.; Harris, M. K.; Wilmarth, W. R.; Calloway, T. B.

    2005-03-23

    In order to accelerate waste treatment and disposal of Hanford tank waste by 2028, the Department of Energy (DOE) and CH2M Hill Hanford Group (CHG), Inc. are evaluating alternative technologies which will be used in conjunction with the Waste Treatment Plant (WTP) to safely pretreat and immobilize the tank waste. Several technologies (Bulk Vitrification and Steam Reforming) are currently being evaluated for immobilizing the pretreated waste. Since the WTP does not have sufficient capacity to pretreat all the waste going to supplemental treatment by the 2028 milestone, two technologies (Selective Dissolution and Fractional Crystallization) are being considered for pretreatment of salt waste. The scope of this task was to: (1) evaluate the recent Savannah River Site (SRS) Tank 41 dissolution campaign and other literature to provide a more complete understanding of selective dissolution, (2) provide an update on the progress of salt dissolution and modeling activities at SRS, (3) investigate SRS experience and outside literature sources on industrial equipment and experimental results of previous fractional crystallization processes, and (4) evaluate recent Hanford AP104 boildown experiments and modeling results and recommend enhancements to the Environmental Simulation Program (ESP) to improve its predictive capabilities. This report provides a summary of this work and suggested recommendations.

  17. Vitrification of mouse MII oocytes: Developmental competency using paclitaxel.

    Science.gov (United States)

    Fesahat, Farzaneh; Faramarzi, Azita; Khoradmehr, Arezoo; Omidi, Marjan; Anbari, Fatemeh; Khalili, Mohammad Ali

    2016-12-01

    Oocyte cryopreservation provides an important alternative for fertility preservation for women who will be treated with cytotoxic drugs. However, it can cause spindle disorganization of microtubules, putting the zygote at risk for aneuploidy. Paclitaxel is known to stabilize the microtubules that constitute the spindle. The aim of this study was to investigate the suitable concentration of paclitaxel for adding to the vitrification media to improve the developmental potential of post-thawed mature oocytes to blastocyst formation in mice. A total of 300 MII oocytes were retrieved from superovulated mice, and were divided into three groups of control, Experimental I, and Experimental II. Oocytes in Experimental I and Experimental II were cryopreserved in the presence of 0.5μM or 1μM of paclitaxel in vitrification media, respectively. After thawing, all oocytes were incubated in G-IVF medium for 1 hour. From each group,12 oocytes were selected for viability evaluation by Hoechst/propidium iodide nuclear staining. Standard in vitro fertilization was performed on the rest of the oocytes and embryo development was followed to the blastocyst stage. Fertilization rate was not significantly different between the three groups. However, the cleavage rate (55%) in Experimental II group was significantly lower compared to Experimental I (88%) and control groups (83%). There was a detectable difference between the three groups at the blastocyst rate (Experimental I and control groups, p = 0.004; Experimental II vs. control and Experimental I, p < 0.001). The highest rates of parthenogenesis and arrest were in Experimental II (16% and 21%, respectively) compared with control (6% and 5%, respectively) and Experimental I (5% and 3%, respectively). There was also a significant decrease in viability rate of oocytes in Experimental II compared to the other groups. A high concentration of paclitaxel, an anticancer drug, interrupted the mouse oocyte competency when supplemented to

  18. Corrosion of Metal Inclusions In Bulk Vitrification Waste Packages

    Energy Technology Data Exchange (ETDEWEB)

    Bacon, Diana H.; Pierce, Eric M.; Wellman, Dawn M.; Strachan, Denis M.; Josephson, Gary B.

    2006-07-31

    The primary purpose of the work reported here is to analyze the potential effect of the release of technetium (Tc) from metal inclusions in bulk vitrification waste packages once they are placed in the Integrated Disposal Facility (IDF). As part of the strategy for immobilizing waste from the underground tanks at Hanford, selected wastes will be immobilized using bulk vitrification. During analyses of the glass produced in engineering-scale tests, metal inclusions were found in the glass product. This report contains the results from experiments designed to quantify the corrosion rates of metal inclusions found in the glass product from AMEC Test ES-32B and simulations designed to compare the rate of Tc release from the metal inclusions to the release of Tc from glass produced with the bulk vitrification process. In the simulations, the Tc in the metal inclusions was assumed to be released congruently during metal corrosion as soluble TcO4-. The experimental results and modeling calculations show that the metal corrosion rate will, under all conceivable conditions at the IDF, be dominated by the presence of the passivating layer and corrosion products on the metal particles. As a result, the release of Tc from the metal particles at the surfaces of fractures in the glass releases at a rate similar to the Tc present as a soluble salt. The release of the remaining Tc in the metal is controlled by the dissolution of the glass matrix. To summarize, the release of 99Tc from the BV glass within precipitated Fe is directly proportional to the diameter of the Fe particles and to the amount of precipitated Fe. However, the main contribution to the Tc release from the iron particles is over the same time period as the release of the soluble Tc salt. For the base case used in this study (0.48 mass% of 0.5 mm diameter metal particles homogeneously distributed in the BV glass), the release of 99Tc from the metal is approximately the same as the release from 0.3 mass% soluble Tc

  19. The solubilities of significant organic compounds in HLW tank supernate solutions -- FY 1995 progress report

    Energy Technology Data Exchange (ETDEWEB)

    Barney, G.S.

    1996-04-26

    At the Hanford Site organic compounds were measured in tank supernate simulant solutions during FY 1995. This solubility information will be used to determine if these organic salts could exist in solid phases (saltcake or sludges) in the waste where they might react violently with the nitrate or nitrite salts present in the tanks. Solubilities of sodium glycolate, succinate, and caproate salts; iron and aluminum and butylphosphate salts; and aluminum oxalate were measured in simulated waste supernate solutions at 25 {degree}C, 30 {degree}C, 40 {degree}C, and 50 {degree}C. The organic compounds were selected because they are expected to exist in relatively high concentrations in the tanks. The solubilities of sodium glycolate, succinate, caproate, and butylphosphate in HLW tank supernate solutions were high over the temperature and sodium hydroxide concentration ranges expected in the tanks. High solubilities will prevent solid sodium salts of these organic acids from precipitating from tank supernate solutions. The total organic carbon concentrations (YOC) of actual tank supernates are generally much lower than the TOC ranges for simulated supernate solutions saturated (at the solubility limit) with the organic salts. This is so even if all the dissolved carbon in a given tank and supernate is due to only one of these eight soluble compounds (an unlikely situation). Metal ion complexes of and butylphosphate and oxalate in supernate solutions were not stable in the presence of the hydroxide concentrations expected in most tanks. Iron and aluminum dibutylphosphate compounds reacted with hydroxide to form soluble sodium dibutylphosphate and precipitated iron and aluminum hydroxides. Aluminum oxalate complexes were also not stable in the basic simulated supernate solutions. Solubilities of all the organic salts decrease with increasing sodium hydroxide concentration because of the common ion effect of Na+. Increasing temperatures raised the solubilities of the organic

  20. Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification

    National Research Council Canada - National Science Library

    Melissa Meneghel; Priscila Chediek Dall’Acqua; Marcela Ambrogi; Beatriz C.S. Leão; Nathália A.S. Rocha-Frigoni; Gisele Z. Mingoti

    The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification...

  1. Viability of zebrafish (Danio rerio) ovarian follicles after vitrification in a metal container.

    OpenAIRE

    MARQUES, L.S.; Bos-Mikich, A.; Godoy, L.C.; Silva,L.A; Maschio, D; Zhang, Tiantian; Streit, D.P.

    2015-01-01

    Cryopreservation of ovarian tissue has been studied for female germline preservation of farm animals and endangered mammalian species. However, there are relatively few reports on cryopreservation of fish ovarian tissue and especially using vitrification approach. Previous studies of our group has shown that the use of a metal container for the cryopreservation of bovine ovarian fragments results in good primordial and primary follicle morphological integrity after vitrification. The aim of t...

  2. Effect of macromolecules in solutions for vitrification of mature bovine oocytes.

    Science.gov (United States)

    Checura, C M; Seidel, G E

    2007-03-15

    This study was designed to evaluate vitrification procedures for in vitro matured bovine oocytes for efficient blastocyst production after warming, IVF and culture. A second goal was to replace serum as the macromolecular component of the vitrification solution, without compromising efficacy. The first experiment compared two containers, open pulled straws (OPS) versus cryoloops, and two vitrification protocols: short equilibration (H-TCM-199+10% EG+10% DMSO+20% FCS for 30s, followed by H-TCM-199+20% EG+20% DMSO+20% FCS+0.48M galactose for 20s) versus long equilibration (H-TCM-199+3% EG+20% FCS for 10min, followed by H-TCM-199+31% EG+20% FCS+1M galactose for 20s). Subsequent experiments used only cryoloops and the short equilibration protocol to evaluate the effect of replacing FCS with defined macromolecules (BSA, Ficoll, PVP, and PVA) in vitrification solutions. Cryoloops were superior to OPS for vitrification of oocytes as determined by blastocyst production (Pvitrification protocols gave similar results. The presence of macromolecules in vitrification solutions for bovine oocytes was necessary for acceptable post-warming developmental capacity; 20% FCS, 1% and 2% BSA, 6% and 18% Ficoll, 6% and 20% PVP, 1% PVA, and the combinations of 18% Ficoll+1% BSA, and 6% PVP+1% BSA provided similar protection during vitrification of oocytes; development ranged from 14.8% to 23.0% blastocysts/oocyte, which was not different (P>0.05) from non-vitrified controls (26.9-34.0% blastocysts/oocyte). Too much (6%) and too little (0.3%) BSA, and 0.3% PVA for vitrification resulted in lower blastocyst production (P<0.05) relative to unvitrified oocytes.

  3. Cryopreservation of bovine oocytes: is cryoloop vitrification the future to preserving the female gamete?

    OpenAIRE

    Mavrides, Andreas; Morroll, David

    2002-01-01

    International audience; The cryoloop is a technique where a thin nylon loop is used to suspend a film of cryoprotectant containing the oocytes and directly immersing them in liquid nitrogen. 508 bovine oocytes were collected, of these 351 were cryopreserved by slow freezing using standard straws or a new vitrification method using our self-constructed cryoloops and the remainder were controls. After thawing, the oocytes were inseminated by ICSI or standard IVF. The cryoloop vitrification meth...

  4. LFCM (liquid-fed ceramic melter) vitrification technology: Quarterly progress report, January--March 1987

    Energy Technology Data Exchange (ETDEWEB)

    Brouns, R. A.; Allen, C. R.; Powell, J. A. (comps.)

    1988-05-01

    This report is compiled by the Nuclear Waste Treatment Program and the Hanford Waste Vitrification Program at Pacific Northwest Laboratory to describe the progress in developing, testing, applying and documenting liquid-fed ceramic melter vitrification technology. Progress in the following technical subject areas during the second quarter of FY 1987 is discussed: melting process chemistry and glass development, feed preparation and transfer systems, melter systems, canister filling and handling systems, and process/product modeling. 23 refs., 14 figs., 10 tabs.

  5. Vitrification of a monatomic simple liquid in two dimensions

    Science.gov (United States)

    Odagaki, Takashi; Mizuguchi, Tomoko

    2011-03-01

    We investigate vitrification and crystallization process of a monatomic system by molecular dynamics simulation, where atoms interact via Lennard-Jones-Gauss potential. We first determine the time-temperature-transformation diagram by observing the crystallization time of the rapidly quenched state from the melt. The crystallization time becomes shortest at a certain temperature T*. The glassy state at low temperatures is shown to be fairly long-lived. In order to examine atomic mechanism of the crystallization, we introduce a modified incoherent intermediate scattering function which measures the structural correlation to a target structure. We show that the crystallization above and below T* take different paths. We also determine the free energy landscape (FEL) and show that the atomic dynamics is consistent with the FEL picture of the glass transition. This work was partially supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture.

  6. Behavior of technetium in nuclear waste vitrification processes.

    Science.gov (United States)

    Pegg, Ian L

    Nearly 100 tests were performed with prototypical melters and off-gas system components to investigate the extents to which technetium is incorporated into the glass melt, partitioned to the off-gas stream, and captured by the off-gas treatment system components during waste vitrification. The tests employed several simulants, spiked with 99m Tc and Re (a potential surrogate), of the low activity waste separated from nuclear wastes in storage in the Hanford tanks, which is planned for immobilization in borosilicate glass. Single-pass technetium retention averaged about 35 % and increased significantly with recycle of the off-gas treatment fluids. The fraction escaping the recycle loop was very small.

  7. Vitrification of bovine matured oocytes and blastocysts in a paper container.

    Science.gov (United States)

    Paul, Ashit Kumar; Liang, Yuanyuan; Srirattana, Kanokwan; Nagai, Takashi; Parnpai, Rangsun

    2017-10-10

    In the present study, we aimed to determine the applicability of a paper container for the vitrification of in vitro matured (IVM) bovine oocytes. In experiment 1, IVM oocytes were exposed to vitrification solution (20% dimethylsulfoxide (DMSO), 20% ethylene glycol (EG), and 5 mol/L sucrose), using a two-step method, for 30 s; loaded onto either a paper container or Cryotop; and stored in liquid nitrogen. No significant difference (P vitrification was observed between the paper container and Cryotop. In experiment 2, IVM oocytes were exposed to either a two- or three-step vitrification solution. The three-step vitrification solution was not significantly different from the two-step solution in terms of oocyte survival, cleavage and blastocyst rates. In experiment 3, in vitro produced blastocysts were graded according to the manual of the International Embryo Transfer Society (grades 1 and 2) and vitrified using the two- and three-step methods. For grade 2 blastocysts, the three-step method showed significantly higher (P vitrification. © 2017 Japanese Society of Animal Science.

  8. Developmental competence and gene expression of immature oocytes following liquid helium vitrification in bovine.

    Science.gov (United States)

    Chen, Jun-Yi; Li, Xiao-Xia; Xu, Ya-Kun; Wu, Hua; Zheng, Jun-Jun; Yu, Xue-Li

    2014-12-01

    The objective of this study was to develop an effective ultra-rapid vitrification method and evaluate its effect on maturation, developmental competence and development-related gene expression in bovine immature oocytes. Bovine cumulus oocyte complexes were randomly allocated into three groups: (1) controls, (2) liquid nitrogen vitrification, and (3) liquid helium vitrification. Oocytes were vitrified and then warmed, the percentage of morphologically normal oocytes in liquid helium group (89.0%) was significantly higher (Pvitrification had higher cleavage and blastocyst rates (41.1% and 10.0%) than that of liquid nitrogen vitrification (33.0% and 4.5%; Pvitrification. Expression of GDF9 and BAX in the liquid helium vitrification group was not significantly different from that of the control, however there were significant differences between the liquid nitrogen vitrification group and control. In conclusion, it was feasible to use liquid helium for vitrifying bovine immature oocytes. There existed an association between the compromised developmental competence and the altered expression levels of these genes for the vitrified oocytes. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. In Vitro Maturation and Embryo Development to blastocyst Mouse Germinal Vesicle Oocytes after Vitrification

    Directory of Open Access Journals (Sweden)

    M Nikseresht

    2013-05-01

    Full Text Available Abstract Background & aim: Vitrification is a simple and ultra rapid technique for the conservation of fertility. Improving pregnancy rate associate with the use of cryopreserved oocytes would be an important advanced in human assisted reproductive technology (ART. The purpose of this study was to evaluate survival, oocytes maturation and embryo development to the blastocyst stage after vitrification of oocytes germinal vesicle-stage and multi stage Methods: In the present experimental study, germinal vesicle oocytes with or without cumulus cells were transferred to vitrification solution containing 30% (v/v ethylene glycol, 18% (w/v Ficoll-70, and 0.3 M sucrose, either by single step or in a step-wise way. After vitrification and storage in liquid nitrogen, the oocytes were thawed and washed twice in culture medium TCM119, and then subjected to in vitro maturation, fertilization, and culture. Data analysis was performed by using One-way variance and Tukey tests. Results: Oocytes survival, metaphase 2 stage oocyte maturation, fertilization and embryo formed blastocyst in vitrification methods multistage were significantly higher than the single step procedure (P<0/05 Conclusion: The Germinal vesicle stage oocytes vitrified with cumulus cells and stepwise procedure had positive effect on the survival, maturation and developmental rate on blastocyst compared to oocytes without cumulus cell and single step procedure. Key words: Germinal Vesicle Oocyte, Blastocyst, Vitrification, Ethylene glycol

  10. Cryopreservation of chicken primordial germ cells by vitrification and slow freezing: A comparative study.

    Science.gov (United States)

    Tonus, C; Connan, D; Waroux, O; Vandenhove, B; Wayet, J; Gillet, L; Desmecht, D; Antoine, N; Ectors, F J; Grobet, L

    2017-01-15

    In the present study, we compare a classical slow freezing (SLF) method and an aseptic vitrification (Vitrif) technique to cryopreserve a stable primordial germ cell (PGCs) line issued from the Ardennaise chicken breed. Viability immediately after warming was close to 80% and did not differ between the two cryopreservation methods. Proliferation tended to be slower for both cryopreservation methods compared with controls, but the difference was significant only for Vitrif. No difference was found between the two methods after flow cytometry analysis of stage-specific embryonic antigen-1 expression and reverse transcription-polymerase chain reaction on several factors related to PGC phenotype. After 1 week in culture, all cryopreserved cells reached controls' main morphologic and expanding (viability/proliferation) features. However, SLF generated more unwanted cells clusters than Vitrif. After injection of the PGCs into recipient embryos, vitrified PGCs reported a clear, yet not significant, tendency to colonize the gonad at a higher rate than slow frozen PGCs. SLF in cryovials remains simple, inexpensive, and less technically demanding than Vitrif. Nevertheless, the intrinsic advantages of our aseptic Vitrif method and the present study suggest that this should be considered as safer than classical SLF for cryopreserving chicken PGCs. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Good Preservation of Stromal Cells and No Apoptosis in Human Ovarian Tissue after Vitrification

    Directory of Open Access Journals (Sweden)

    Raffaella Fabbri

    2014-01-01

    Full Text Available The aim of this study was to develop a vitrification procedure for human ovarian tissue cryopreservation in order to better preserve the ovarian tissue. Large size samples of ovarian tissue retrieved from 15 female-to-male transgender subjects (18–38 years were vitrified using two solutions (containing propylene glycol, ethylene glycol, and sucrose at different concentrations in an open system. Light microscopy, transmission electron microscopy, and TUNEL assay were applied to evaluate the efficiency of the vitrification protocol. After vitrification/warming, light microscopy showed oocyte nucleus with slightly thickened chromatin and irregular shape, while granulosa and stromal cells appeared well preserved. Transmission electron microscopy showed oocytes with slightly irregular nuclear shape and finely dispersed chromatin. Clear vacuoles and alterations in cellular organelles were seen in the oocyte cytoplasm. Stromal cells had a moderately dispersed chromatin and homogeneous cytoplasm with slight vacuolization. TUNEL assay revealed the lack of apoptosis induction by vitrification in all ovarian cell types. In conclusion after vitrification/warming the stromal compartment maintained morphological and ultrastructural features similar to fresh tissue, while the oocyte cytoplasm was slightly damaged. Although these data are encouraging, further studies are necessary and essential to optimize vitrification procedure.

  12. Good preservation of stromal cells and no apoptosis in human ovarian tissue after vitrification.

    Science.gov (United States)

    Fabbri, Raffaella; Vicenti, Rossella; Macciocca, Maria; Pasquinelli, Gianandrea; Paradisi, Roberto; Battaglia, Cesare; Martino, Nicola Antonio; Venturoli, Stefano

    2014-01-01

    The aim of this study was to develop a vitrification procedure for human ovarian tissue cryopreservation in order to better preserve the ovarian tissue. Large size samples of ovarian tissue retrieved from 15 female-to-male transgender subjects (18-38 years) were vitrified using two solutions (containing propylene glycol, ethylene glycol, and sucrose at different concentrations) in an open system. Light microscopy, transmission electron microscopy, and TUNEL assay were applied to evaluate the efficiency of the vitrification protocol. After vitrification/warming, light microscopy showed oocyte nucleus with slightly thickened chromatin and irregular shape, while granulosa and stromal cells appeared well preserved. Transmission electron microscopy showed oocytes with slightly irregular nuclear shape and finely dispersed chromatin. Clear vacuoles and alterations in cellular organelles were seen in the oocyte cytoplasm. Stromal cells had a moderately dispersed chromatin and homogeneous cytoplasm with slight vacuolization. TUNEL assay revealed the lack of apoptosis induction by vitrification in all ovarian cell types. In conclusion after vitrification/warming the stromal compartment maintained morphological and ultrastructural features similar to fresh tissue, while the oocyte cytoplasm was slightly damaged. Although these data are encouraging, further studies are necessary and essential to optimize vitrification procedure.

  13. Need for USA high level waste (HLW) alternate geological repository (AGR) and for a different methodology to enhance its acceptance

    Energy Technology Data Exchange (ETDEWEB)

    Levy, Salomon, E-mail: slevy112@aol.co [3425 South Bascom Avenue, Suite 225, Campbell, CA 95008 (United States)

    2010-10-15

    In early February 2010, the administration stopped work and withdrew the Department of Energy (DOE) application for a construction permit for the Yucca Mountain geological repository from the Nuclear Regulatory Commission (NRC). Also, a 'blue ribbon' Commission was appointed to explore alternatives for storage, processing, and disposal, including evaluation of advanced fuel cycles and to provide a final report in 24 months. That decision, however, failed to recognize that: (1) the U.S. will need an early alternate geological repository (AGR) for its HLW irrespective of the findings of the 'blue ribbon' Commission; (2) the once-through spent fuel inventory from commercial nuclear power reactors will continue to rise and so will the damages against the government for its failure to remove spent fuel from reactors sites, as specified in contracts; (3) there are prepackaged DOE and nuclear weapons HLW ready for shipment to a repository which must be taken into account because of government penalties for failure to do so; (4) the current Nuclear Waste Policy Act (NWPA) needs to be modified to allow the early search and approval of Alternate Geological Repository (AGR) and for an interim centralized HLW storage facility to reduce government liabilities; and (5) the methodology used to license Yucca Mountain needs to undergo serious modifications, including a different non-politicized management and siting credo. This paper reviews and discusses all the preceding shortcomings and proposes significant changes to pursue AGR as soon as possible and to get site approval by the NRC first under a formal, stepwise, well-structured risk-informed decision approach as recommended.

  14. Monotone viable trajectories for functional differential inclusions

    Science.gov (United States)

    Haddad, Georges

    This paper is a study on functional differential inclusions with memory which represent the multivalued version of retarded functional differential equations. The main result gives a necessary and sufficient equations. The main result gives a necessary and sufficient condition ensuring the existence of viable trajectories; that means trajectories remaining in a given nonempty closed convex set defined by given constraints the system must satisfy to be viable. Some motivations for this paper can be found in control theory where F( t, φ) = { f( t, φ, u)} uɛU is the set of possible velocities of the system at time t, depending on the past history represented by the function φ and on a control u ranging over a set U of controls. Other motivations can be found in planning procedures in microeconomics and in biological evolutions where problems with memory do effectively appear in a multivalued version. All these models require viability constraints represented by a closed convex set.

  15. Status of Progress Made Toward Safety Analysis and Technical Site Evaluations for DOE Managed HLW and SNF.

    Energy Technology Data Exchange (ETDEWEB)

    Sevougian, S. David [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Stein, Emily [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Gross, Michael B [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Hammond, Glenn Edward [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Frederick, Jennifer M [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Mariner, Paul [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-11-01

    The Spent Fuel and Waste Science and Technology (SFWST) Campaign of the U.S. Department of Energy (DOE) Office of Nuclear Energy (NE) is conducting research and development (R&D) on generic deep geologic disposal systems (i.e., repositories). This report describes specific activities in FY 2016 associated with the development of a Defense Waste Repository (DWR)a for the permanent disposal of a portion of the HLW and SNF derived from national defense and research and development (R&D) activities of the DOE.

  16. Dose Calculations for the Co-Disposal WP-of HLW-Glass and the Triga SNF

    Energy Technology Data Exchange (ETDEWEB)

    G. Radulescu

    1999-08-02

    This calculation is prepared by the Monitored Geologic Repository (MGR) Waste Package Operations (WPO). The purpose of this calculation is to determine the surface dose rates of a codisposal waste package (WP) containing a centrally located Department of Energy (DOE) standardized 18-in. spent nuclear fuel (SNF) canister, loaded with the TRIGA (Training, Research, Isotopes, General Atomics) SNF. This canister is surrounded by five 3-m long canisters, loaded with Savannah River Site (SRS) high-level waste (HLW) glass. The results are to support the WP design and radiological analyses.

  17. 'Personalisation' of droplet-vitrification protocols for plant cells: a systematic approach to optimising chemical and osmotic effects.

    Science.gov (United States)

    Kim, Haeng-Hoon; Lee, Sheong-Chun

    2012-01-01

    Although an appropriate cryopreservation protocol is a prerequisite for basic studies and large-scale implementation as well as further cryopreservation studies, the process relies on trial and error. Among the vitrification-based cryopreservation techniques, droplet-vitrification produces high post-cryopreservation recovery. However, the protocol itself cannot solve the problems engaged in plant cryopreservation, prominently due to dehydration with cytotoxic vitrification solutions. This paper proposes a set of treatments to develop droplet-vitrification using a standard procedure associated with additional treatments and alternative vitrification solutions. The proposed standard protocol consists of a progressive preculture with 0.3 M sucrose for 31 h and with 0.7 M for 17 h, loading with vitrification solution C4-35% (17.5 percent glycerol + 17.5 percent sucrose, w/v) for 20 to 40 min, dehydration with vitrification solutions A3-90 percent (37.5 percent glycerol + 15% DMSO + 15 percent EG + 22.5 percent sucrose) for 10 to 30 min or B1-100 percent (PVS3) for 40 to 120 min at room temperature, cooling the samples using aluminum foil strips, rewarming by plunging into pre-heated (40 degree C) unloading solution (0.8 M sucrose) and further unloading for 20 to 60 min, depending on size and permeability of the materials. Using this systematic approach we can identify whether the material is tolerant or sensitive to chemical toxicity and to the osmotic stress of dehydration with vitrification solutions, thus revealing which is the main barrier in solution-based vitrification methods. Based on the sensitivity of samples we can design a droplet-vitrification procedure, i.e. preculture, loading, dehydration with vitrification solutions, cooling and rewarming. Using this approach, the development of appropriate droplet-vitrification protocol is facilitated.

  18. Oocyte maturation, embryo development and gene expression following two different methods of bovine cumulus-oocyte complexes vitrification.

    Science.gov (United States)

    Azari, Mehdi; Kafi, Mojtaba; Ebrahimi, Bita; Fatehi, Roya; Jamalzadeh, Mahboobeh

    2017-03-01

    To examine the maturational competence, embryo development and expression of genes involved in oocyte maturation and cumulus expansion (GDF9, BMP15, HAS2, TNFAIP6, FGF17 and FSHr) following two standard methods of bovine COCs vitrification. Bovine cumulus-oocyte complexes (COCs) were aspirated from slaughtered ovaries and then distributed into three groups: non-vitrified COCs (control), vitrification 1 group (V1); vitrification was performed by 15% ethylene glycol (EG) and 15% DMSO in holding media (TCM-199 with 20% FCS); and vitrification 2 group (V2); vitrification was performed by 40% EG in holding media. After vitrification, COCs were warmed in two steps and cultured and then evaluated for nuclear maturation, embryo development and gene expressions. The mean (±SD) percentages of nuclear maturation and blastocyst/cleaved were higher in control group (79.5 ± 8.0 and 31.0 ± 5.1%) than the V1 (34.8 ± 9.1 and 4.4 ± 5.1%) and V2 (47.8 ± 11.7 and 7.1 ± 5.8%) groups (P vitrification groups (P vitrification procedure and conditions. Using EG alone for vitrification of bovine immature COCs, resulted in higher expression of GDF9, BMP15 and production of more in vitro matured and cleaved oocytes.

  19. FINAL REPORT DM1200 TESTS WITH AZ 101 HLW SIMULANTS VSL-03R3800-4 REV 0 2/17/04

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; BARDAKCI T; D' ANGELO NA; GONG W; KOT WK; PEGG IL

    2011-12-29

    This report documents melter and off-gas performance results obtained on the DM 1200 HLW Pilot Melter during processing of simulated HLW AZ-101 feed. The principal objectives of the DM1200 melter testing were to determine the achievable glass production rates for simulated HLW AZ-101 feed; determine the effect of bubbling rate and feed solids content on production rate; characterize melter off-gas emissions; characterize the performance of the prototypical off-gas system components as well as their integrated performance; characterize the feed, glass product, and off-gas effluents; and to perform pre- and post-test inspections of system components. The test objectives (including test success criteria), along with how they were met, are outlined in a table.

  20. Solubilities of significant compounds in HLW tank supernate solutions - FY 1996 progress report

    Energy Technology Data Exchange (ETDEWEB)

    Barney, G.S.

    1996-09-30

    The solubilities of two sodium salts of organic acids that are thought to exist in high-level waste at the Hanford Site were measured in tank supernate simulant solutions during FY1996 This solubility information will be used to determine if these organic salts could exist in solid phases (saltcake or sludges) in the waste where they might react violently with the nitrate or nitrite salts present in the tanks. Solubilities of sodium butyrate and trisodium N-(2-hydroxyethyl)ethylenediaminetriacetate were measured in simulated waste supernate solutions at 25 {degrees}C, 30 {degrees}C, 40 {degrees}C, and 50 {degrees}C. The organic compounds were selected because they are expected to exist in relatively high concentrations in the tanks. Two types of tank supernate simulants were used - a 4.O M sodium nitrate - 0.97 M sodium nitrite solution with sodium hydroxide concentrations ranging from O.00003 M to 2.O M and a 2.O M sodium nitrite solution saturated with crystalline sodium nitrate with sodium hydroxide concentrations ranging from 0.1 M to 2. 0 M. The solubilities of sodium butyrate and trisodium N-(2-hydroxyethyl)ethylene- diaminetriacetate in both types of HLW tank supernate solutions were high over the temperature and sodium hydroxide concentration ranges expected in the tanks. The solubilities of these compounds are similar (in terms of total organic carbon) to sodium glycolate, succinate, caproate, dibutylphosphate, citrate, formate, ethylenediaminetetraacetate, and nitrilotriacetate which were measured previously. High solubilities will prevent solid sodium salts of these organic acids from precipitating from tank supernate solutions. The total organic carbon concentrations (TOC) of actual tank supernates are generaly much lower than the TOC ranges for the simulated supernate solutions saturated (at the solubility limit) with the organic salts. This is true even if all the dissolved carbon in a given tank supernate is due to only one of these eight soluble

  1. Site selection process for a HLW geological repository in France. A convergence approach

    Energy Technology Data Exchange (ETDEWEB)

    Solente, N.; Ouzounian, G.; Miguez, R.; Tison, J-L. [ANDRA Agence Nationale pour la Gestion des Dechets Radioactifs, Chatenay-Malabry (France)

    2011-07-01

    On December 1991, the French National Assembly passed the French Waste Management Research Act, authorizing and initiating a 15 year research program along three options for HLW long term solution: separation and/or transmutation, long-term storage, and geologic disposal. On June 2006, the 'Planning Act on the sustainable management of radioactive materials and waste' sets a new framework and new aims to the above mentioned options. This paper deals only with the geologic disposal research program. In a step by step approach, this program has been broken down into three phases, each having intermediate objectives: site selection for an Underground research Laboratory (URL), disposal feasibility demonstration, reversible disposal design. The first step of the research program aimed at URL site selection. From 1994 to 1996, Andra carried out geological characterization surveys in four French districts, leading to the Request for Licensing and Operation of laboratory facilities on three sites. During this phase, boreholes, 2D seismic campaigns and outcrops geologic studies were the main sources of data. The result was the selection of Bure area, the most suitable site for the implementation of an underground laboratory. Main results on Bure URL will be presented in the paper. In the second phase the research program targeted the safety and technical feasibility of a reversible disposal site, located in Meuse or Haute Marne districts, as selected by the government in 1998. Andra conducted geologic survey during the URL shaft sinking and experiments in drifts at depths of 445 and 490 m. This program allowed consolidating the knowledge already acquired: geological environment, stability of the rock and the regional geology, and containment properties. The 2005 Progress Report presents the results of this phase. The main conclusion is that a potential disposal facility may be safely constructed over a zone with geological characteristics similar to those

  2. Air-spore in Cartagena, Spain: viable and non-viable sampling methods.

    Science.gov (United States)

    Elvira-Rendueles, Belen; Moreno, Jose; Garcia-Sanchez, Antonio; Vergara, Nuria; Martinez-Garcia, Maria Jose; Moreno-Grau, Stella

    2013-01-01

    In the presented study the airborne fungal spores of the semiarid city of Cartagena, Spain, are identified and quantified by means of viable or non-viable sampling methods. Airborne fungal samples were collected simultaneously using a filtration method and a pollen and particle sampler based on the Hirst methodology. This information is very useful for elucidating geographical patterns of hay fever and asthma. The qualitative results showed that when the non-viable methodology was employed, Cladosporium, Ustilago, and Alternaria were the most abundant spores identified in the atmosphere of Cartagena, while the viable methodology showed that the most abundant taxa were: Cladosporium, Penicillium, Aspergillus and Alternaria. The quantitative results of airborne fungal spores identified by the Hirst-type air sampler (non-viable method), showed that Deuteromycetes represented 74% of total annual spore counts, Cladosporium being the major component of the fungal spectrum (62.2%), followed by Alternaria (5.3%), and Stemphylium (1.3%). The Basidiomycetes group represented 18.9% of total annual spore counts, Ustilago (7.1%) being the most representative taxon of this group and the second most abundant spore type. Ascomycetes accounted for 6.9%, Nectria (2.3%) being the principal taxon. Oomycetes (0.2%) and Zygomycestes and Myxomycestes (0.06%) were scarce. The prevailing species define our bioaerosol as typical of dry air. The viable methodology was better at identifying small hyaline spores and allowed for the discrimination of the genus of some spore types. However, non-viable methods revealed the richness of fungal types present in the bioaerosol. Thus, the use of both methodologies provides a more comprehensive characterization of the spore profile.

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

    Science.gov (United States)

    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 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 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.

  4. Successful application of the strategy of blastocyst biopsy, vitrification, whole genome amplification, and thawed embryo transfer for preimplantation genetic diagnosis of neurofibromatosis type 1

    Directory of Open Access Journals (Sweden)

    Yi-Lin Chen

    2011-03-01

    Conclusion: We first demonstrate successful application of blastocyst biopsy, vitrification, WGA, and thawed embryo transfer for PGD of a monogenic disease. Vitrification of blastocysts after biopsy permits sufficient time for shipment of samples and operation of molecular diagnosis.

  5. Parejas viables que perduran en el tiempo

    OpenAIRE

    Juan José Cuervo Rodríguez

    2013-01-01

    El presente artículo científico presenta resultados del proceso llevado a cabo en el proyecto de investigación docente "Mecanismos de autorregulación en parejas viables que perduran en el tiempo". Se soporta en una mirada compleja de la psicología basada en una epistemología de la construcción. En el ámbito metodológico, se inscribe en los estudios de terapia familiar desde una perspectiva de la comunicación humana como un todo integrado. Participaron nueve parejas. Los criterios de inclusión...

  6. Enzymatic isolation of viable human odontoblasts.

    Science.gov (United States)

    Cuffaro, H M; Pääkkönen, V; Tjäderhane, L

    2016-05-01

    To improve an enzymatic method previously used for isolation of rat odontoblasts to isolate viable mature human odontoblasts. Collagenase I, collagenase I/hyaluronidase mixture and hyaluronidase were used to extract mature human odontoblasts from the pulp chamber. Detachment of odontoblasts from dentine was determined with field emission scanning electron microscopy (FESEM) and to analyse the significance of differences in tubular diameter, and the t-test was used. MTT-reaction was used to analyse cell viability, and nonparametric Kruskal-Wallis and Mann-Whitney post hoc tests were used to analyse the data. Immunofluorescent staining of dentine sialoprotein (DSP), aquaporin-4 (AQP4) and matrix metalloproteinase-20 (MMP-20) and quantitative PCR (qPCR) of dentine sialophosphoprotein (DSPP) were used to confirm the odontoblastic nature of the cells. MTT-reaction and FESEM demonstrated collagenase I/hyaluronidase resulted in more effective detachment and higher viability than collagenase I alone. Hyaluronidase alone was not able to detach odontoblasts. Immunofluorescence revealed the typical odontoblastic-morphology with one process, and DSP, AQP4 and MMP-20 were detected. Quantitative PCR of DSPP confirmed that the isolated cells expressed this odontoblast-specific gene. The isolation of viable human odontoblasts was successful. The cells demonstrated morphology typical for odontoblasts and expressed characteristic odontoblast-type genes and proteins. This method will enable new approaches, such as apoptosis analysis, for studies using fully differentiated odontoblasts. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  7. Structural Changes in Cattle Immature Oocytes Subjected to Slow Freezing and Vitrification

    Directory of Open Access Journals (Sweden)

    H. Wahid*, M. Thein1, E.A. El-Hafez2, M.O. Abas3, K. Mohd Azam4, O. Fauziah5, Y. Rosnina and H. Hajarian

    2012-05-01

    Full Text Available This study was conducted to evaluate the effect of different cryopreservation methods (slow-freezing and vitrification on structural changes of bovine immature oocytes. Bovine ovaries were collected from local abattoirs. Cumulus-oocyte-complexes (COCs were retrieved using aspiration method from 2-6 mm follicles. In Experiment 1, selected oocytes were randomly divided into 4 treatment groups namely freezing solution-exposed, frozen-thawed, vitrification solution-exposed and vitrified-thawed and then oocytes abnormalities were examined under a stereomicroscope. In Experiment 2, oocytes were randomly allocated to the same grouping as experiment 1 plus control group. Following freezing or vitrification, all oocytes were fixed in glutaraldehyde and processed for transmission electron microscopy. In experiment 1, there was a higher incidence of abnormalities in the frozen-thawed and vitrified-warmed oocytes compared to those in freezing solution and vitrification solution-exposed groups (P<0.05. In experiment 2, there were marked alterations in the perivitelline space, microvilli and vesicles of frozen-thawed and vitrified-warmed oocytes characterized by loss of elasticity and integrity of cytoplasmic processes and microvilli following cooling and warming. In conclusion, ethylene glycol-based freezing and vitrification solutions are suitable choices for cryopreservation of immature oocytes and most organelles are able to retain their normal morphology following cryopreservation and thawing processes.

  8. Vitrification of bovine preantral follicles with dimethylsulfoxide and sucrose plus α-tocopherol

    Directory of Open Access Journals (Sweden)

    Carolina R. Jimenez

    2016-03-01

    Full Text Available Abstract: The objective of this study was to evaluate the vitrification of bovine preantral follicles with dimethylsulfoxide (D and sucrose (S plus α-tocopherol 5mmol/L (T5 or 10mmol/L (T10 and, evaluate the thawed with minimal essential medium (m with or without sucrose (s. Ovaries of cows were collected from slaughterhouse for the experiment I (n=66 and II (n=51. In the laboratory ovarian fragments were randomly assigned either to fresh control and 8 vitrification treatments (Controle and Dm; Dms, DSm; DSms; DST5m; DST5ms; DST10m; DST10ms. Ovarian fragments were placed in vitrification solution (5 min and immersed in liquid nitrogen (-196°C, after a week, the fragments were thawed and analyzed. In the experiments I, preantral follicles were morphologically observed for histological evaluation, (normal; degenerated and developing of stage. In the experiment II, preantral follicles were mechanically isolated from ovarian tissue and examined with trypan blue, where dead and live corresponded to stained or non-stained. The treatments DSm, DSms and DST10m were effective in preserving the morphology in situ. However, the viability of isolated preantral follicles after vitrification remained high only in treatment DST10m. Thus, DST10m preserves survival rates and morphological integrity during vitrification of bovine preantral follicles.

  9. Effect of bovine ovarian tissue vitrification on the structural preservation of antral follicles.

    Science.gov (United States)

    Faheem, M S; Carvalhais, I; Baron, E; Moreira da Silva, F

    2013-10-01

    This study was performed to evaluate the structural preservation of antral follicles after bovine ovarian tissue vitrification using histological analysis. Ovaries (n = 30) of slaughtered cows were cut into small fragments using a scalpel blade, and the ovarian tissues were randomly assigned to vitrification using 15% dimethyl sulphoxide (DMSO) and 15% ethylene glycol (EG) and fresh tissues (control) groups. For histological evaluations, fresh and post-thawing ovarian tissues were immediately fixed, serially sectioned into 5-μm sections and stained with haematoxylin and eosin (H&E). Nine serial sections per fragment were subjected for morphological assessment. The diameter of the antral follicles was determined and classified into four groups: 1 (≤1 mm), 2 (>1-2 mm), 3 (>2-3 mm) and 4 (>3-4 mm). Then, follicular morphology was evaluated in relation to atresia and categorized into seven grades: Grade A (healthy follicle); Grades B, C and D (early atresia); Grades E and F (moderate atresia); and Grade G (advanced atresia). The results revealed that small diameters of antral follicles (1 and 2 mm) were more susceptible for cryoinjury. The normal follicular morphology (Grade A) was not affected by vitrification throughout follicle diameters. Nevertheless, some damage features were monitored after vitrification. In conclusion, the morphological structure of bovine antral follicles could be successfully preserved by ovarian tissue vitrification. © 2013 Blackwell Verlag GmbH.

  10. Cryopreservation of Thymus cariensis and T. vulgaris shoot tips: comparison of three vitrification-based methods.

    Science.gov (United States)

    Ozudogru, E A; Kaya, E

    2012-01-01

    Thymus is an important genus of the Lamiaceae family, comprising more than 400 perennial aromatic thyme species, which are used extensively for medicinal and culinary purposes. The present study focused on the development of cryopreservation procedures for Thymus vulgaris and T. cariensis, the latter being an endemic and endangered species of Turkey. For cryopreservation of T. vulgaris shoot tips, PVS2-based one-step freezing methods, i.e., PVS2 vitrification, encapsulation-vitrification and droplet-vitrification, were compared. Cold hardening and sucrose preculture were also optimized before the cryopreservation trials. For T. cariensis, a droplet-vitrification method was applied to cold-hardened shoot tips, and after sucrose preculture. In all the methods tested, PVS2 was applied for up to 120 min. The best T. vulgaris cryopreservation was achieved with a droplet-vitrification method, that involved 2-weeks cold hardening of shoot cultures, 48 h preculture of shoot tips on MS medium supplemented with 0.25 M sucrose, and a 90 min PVS2 treatment in droplets. After direct immersion in LN, thawing and plating, 80% of shoot-tips recovered. Post-thaw recovery was significantly lower when the same procedure was applied to T. cariensis shoot tips; however also here 90 min PVS2 treatment produced the highest survival (25 percent) and recovery (25 percent) levels.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Yanzhou Yang

    2015-01-01

    Full Text Available 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.

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

    Science.gov (United States)

    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.

  15. BENEFITS OF VIBRATION ANALYSIS FOR DEVELOPMENT OF EQUIPMENT IN HLW TANKS - 12341

    Energy Technology Data Exchange (ETDEWEB)

    Stefanko, D.; Herbert, J.

    2012-01-10

    Vibration analyses of equipment intended for use in the Savannah River Site (SRS) radioactive liquid waste storage tanks are performed during pre-deployment testing and has been demonstrated to be effective in reducing the life-cycle costs of the equipment. Benefits of using vibration analysis to identify rotating machinery problems prior to deployment in radioactive service will be presented in this paper. Problems encountered at SRS and actions to correct or lessen the severity of the problem are discussed. In short, multi-million dollar cost saving have been realized at SRS as a direct result of vibration analysis on existing equipment. Vibration analysis of equipment prior to installation can potentially reduce inservice failures, and increases reliability. High-level radioactive waste is currently stored in underground carbon steel waste tanks at the United States Department of Energy (DOE) Savannah River Site and at the Hanford Site, WA. Various types of rotating machinery (pumps and separations equipment) are used to manage and retrieve the tank contents. Installation, maintenance, and repair of these pumps and other equipment are expensive. In fact, costs to remove and replace a single pump can be as high as a half million dollars due to requirements for radioactive containment. Problems that lead to in-service maintenance and/or equipment replacement can quickly exceed the initial investment, increase radiological exposure, generate additional waste, and risk contamination of personnel and the work environment. Several different types of equipment are considered in this paper, but pumps provide an initial example for the use of vibration analysis. Long-shaft (45 foot long) and short-shaft (5-10 feet long) equipment arrangements are used for 25-350 horsepower slurry mixing and transfer pumps in the SRS HLW tanks. Each pump has a unique design, operating characteristics and associated costs, sometimes exceeding a million dollars. Vibration data are routinely

  16. Vitrification, a complementary cryopreservation method for Betula pendula Roth.

    Science.gov (United States)

    Ryynänen, Leena; Aronen, Tuija

    2005-10-01

    Cryopreservation--the storage of plant germplasm in liquid nitrogen--provides a modern tool for the conservation of forest genetic resources. It is especially applicable for species in which their micropropagation can be initiated from mature tree buds, e.g., silver birch (Betula pendula Roth), thus enabling the conservation of specific genotypes: endangered elite trees and trees expressing rare, valuable or interesting characteristics. The aim of the present study was to develop a vitrification protocol applicable for the cryostorage of silver birch that avoids the use of expensive sophisticated freezers. The average recovery of vitrified axillary silver birch buds was 71% using a protocol that started with four-week cold hardening of bud-bearing in vitro donor shoots on modified medium under short day conditions. After cold hardening, the excised axillary buds were precultivated on medium containing 0.7 M sucrose for 24 h under the same conditions as during the cold hardening period. Following preculture, the buds were treated with loading solution containing 2M glycerol and 0.4 M sucrose for 20 min at room temperature. Finally, the buds were dehydrated with PVS2 cryoprotectant for 120 min followed by direct immersion in liquid nitrogen. According to the morphology and the RAPD profiles of regenerated plants in the greenhouse, the genetic fidelity of the vitrified birch material seems to have remained unchanged.

  17. Laser-assisted vitrification of large equine embryos.

    Science.gov (United States)

    Scherzer, J; Davis, C; Hurley, D J

    2011-12-01

    The major difficulty in providing the benefits of embryo cryopreservation for equine agriculture is the mismatch between the optimal embryo age for collection from the mare (7-8 days after ovulation was detected) and the optimal age for freezing under current methods (6.5 days after ovulation). To overcome this limitation, we tested a method to enhance penetration of cryopreservative across the capsule and trophoblast of day 7 and 8 embryos combined with rapid freezing by vitrification. Six small embryos (laser system used to create a small opening in the embryonic capsule and trophectoderm. All embryos were vitrified using a CryoLeaf freezing support. After recovery from freezing and embryo transfer, three of four small untreated embryos (300 μm in diameter, 44%) resulted in a vesicle as detected by ultrasonography approximately one week after transfer. However, only one recipient mare was still pregnant on day 23, and she delivered a live foal. Further investigation is required to determine why most of the embryos in this experiment were lost between day 13 and day 23 of gestation. © 2011 Blackwell Verlag GmbH.

  18. Temperature Distribution within a Cold Cap during Nuclear Waste Vitrification.

    Science.gov (United States)

    Dixon, Derek R; Schweiger, Michael J; Riley, Brian J; Pokorny, Richard; Hrma, Pavel

    2015-07-21

    The kinetics of the feed-to-glass conversion affects the waste vitrification rate in an electric glass melter. The primary area of interest in this conversion process is the cold cap, a layer of reacting feed on top of the molten glass. The work presented here provides an experimental determination of the temperature distribution within the cold cap. Because direct measurement of the temperature field within the cold cap is impracticable, an indirect method was developed in which the textural features in a laboratory-made cold cap with a simulated high-level waste feed were mapped as a function of position using optical microscopy, scanning electron microscopy, energy dispersive spectroscopy, and X-ray diffraction. The temperature distribution within the cold cap was established by correlating microstructures of cold-cap regions with heat-treated feed samples of nearly identical structures at known temperatures. This temperature profile was compared with a mathematically simulated profile generated by a cold-cap model that has been developed to assess the rate of glass production in a melter.

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

    Science.gov (United States)

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

  20. Roots of success: cultivating viable community forestry

    Energy Technology Data Exchange (ETDEWEB)

    MacQueen, Duncan

    2009-05-15

    Is community forestry emerging from the shadows? The evidence shows that locally controlled enterprises can be economically viable, and often build on stronger social and environmental foundations than the big private-sector players. Certainly this is an industry in need of a shakeup. Many forests have become flashpoints where agro-industry, large-scale logging concerns and conservation interests clash, while forest-dependent communities are left out in the cold. Meanwhile, governments – driven by concerns over the climate impacts of deforestation – are having to gear up for legal, sustainable forestry production. Community forestry could be crucial to solving many of these challenges. By building on local core capabilities and developing strategic partnerships, they are forging key new business models that could transform the sector.

  1. Enumeration of viable and non-viable larvated Ascaris eggs with quantitative PCR.

    Science.gov (United States)

    Raynal, Maria; Villegas, Eric N; Nelson, Kara L

    2012-12-01

    The goal of this study was to further develop an incubation-quantitative polymerase chain reaction (qPCR) method for quantifying viable Ascaris eggs by characterizing the detection limit and number of template copies per egg, determining the specificity of the method, and testing the method with viable and inactivated larvated eggs. The number of template copies per cell was determined by amplifying DNA from known numbers of eggs at different development stages; the value was estimated to be 32 copies. The specificity of the method was tested against a panel of bacteria, fungi, protozoa and helminths, and no amplification was found with non-target DNA. Finally, fully larvated eggs were inactivated by four different treatments: 254 nm ultraviolet light, 2,000 ppm NH(3)-N at pH 9, moderate heat (48 °C) and high heat (70 °C). Concentrations of treated eggs were measured by direct microscopy and incubation-qPCR. The qPCR signal decreased following all four treatments, and was in general agreement with the decrease in viable eggs determined by microscopy. The incubation-qPCR method for enumerating viable Ascaris eggs is a promising approach that can produce results faster than direct microscopy, and may have benefits for applications such as assessing biosolids.

  2. Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum

    Directory of Open Access Journals (Sweden)

    Ching Giap Tan

    2014-02-01

    Full Text Available The present study was based on the reverse transcription polymerase chain reaction (RT-PCR of the 16S ribosomal nucleic acid (rRNA of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used and the suspensions were stored at different temperatures. The 16S rRNA of M. gallisepticum was detected up to approximately 20–25 h at 37 °C, 22–25 h at 16 °C, and 23–27 h at 4 °C. The test, therefore, could detect viable or recently dead M. gallisepticum (< 20 h. The RT-PCR method was applied during an in vivo study of drug efficacy under experimental conditions, where commercial broiler-breeder eggs were inoculated with M. gallisepticum into the yolk. Hatched chicks that had been inoculated in ovo were treated with Macrolide 1. The method was then applied in a flock of day 0 chicks with naturally acquired vertical transmission of M. gallisepticum, treated with Macrolide 2. Swabs of the respiratory tract were obtained for PCR and RT-PCR evaluations to determine the viability of M. gallisepticum. This study proved that the combination of both PCR and RT-PCR enables detection and differentiation of viable from non-viable M. gallisepticum.

  3. Hanford Waste Vitrification program pilot-scale ceramic melter Test 23

    Energy Technology Data Exchange (ETDEWEB)

    Goles, R.W.; Nakaoka, R.K.

    1990-02-01

    The pilot-scale ceramic melter test, was conducted to determine the vitrification processing characteristics of simulated Hanford Waste Vitrification Plant process slurries and the integrated performance of the melter off-gas treatment system. Simulated melter feed was prepared and processed to produce glass. The vitrification system, achieved an on-stream efficiency of greater than 98%. The melter off-gas treatment system included a film cooler, submerged bed scrubber, demister, high-efficiency mist eliminator, preheater, and high-efficiency particulate air filter (HEPA). Evaluation of the off-gas system included the generation, nature, and capture efficiency of gross particulate, semivolatile, and noncondensible melter products. 17 refs., 48 figs., 61 tabs.

  4. Obesity does not aggravate vitrification injury in mouse embryos: a prospective study

    Directory of Open Access Journals (Sweden)

    Ma Wenhong

    2012-08-01

    Full Text Available Abstract Background Obesity is associated with poor reproductive outcomes, but few reports have examined thawed embryo transfer in obese women. Many studies have shown that increased lipid accumulation aggravates vitrification injury in porcine and bovine embryos, but oocytes of these species have high lipid contents (63 ng and 161 ng, respectively. Almost nothing is known about lipids in human oocytes except that these cells are anecdotally known to be relatively lipid poor. In this regard, human oocytes are considered to be similar to those of the mouse, which contain approximately 4 ng total lipids/oocyte. To date, no available data show the impact of obesity on vitrification in mouse embryos. The aim of this study was to establish a murine model of maternal diet-induced obesity and to characterize the effect of obesity on vitrification by investigating the survival rate and embryo developmental competence after thawing. Methods Prospective comparisons were performed between six–eight-cell embryos from obese and normal-weight mice and between fresh and vitrified embryos. Female C57BL/6 mice were fed standard rodent chow (normal-weight group or a high-fat diet (obese group for 6 weeks. The mice were mated, zygotes were collected from oviducts and cultured for 3 days, and six–eight-cell embryos were then selected to assess lipid content in fresh embryos and to evaluate differences in apoptosis, survival, and development rates in response to vitrification. Results In fresh embryos from obese mice, the lipid content (0.044 vs 0.030, Pvs.9.3%, Pvs. 93.1%, P Conclusions This study demonstrated that differences in survival and developmental rates between embryos from obese and normal-weight mice were eliminated after vitrification. Thus, maternal obesity does not aggravate vitrification injury, but obesity alone greatly impairs pre-implantation embryo survival and development.

  5. An improved vitrification protocol for equine immature oocytes, resulting in a first live foal.

    Science.gov (United States)

    Ortiz-Escribano, N; Bogado Pascottini, O; Woelders, H; Vandenberghe, L; De Schauwer, C; Govaere, J; Van den Abbeel, E; Vullers, T; Ververs, C; Roels, K; Van De Velde, M; Van Soom, A; Smits, K

    2017-08-20

    The success rate for vitrification of immature equine oocytes is low. Although vitrified-warmed oocytes are able to mature, further embryonic development appears to be compromised. The aim of this study was to compare two vitrification protocols, and to examine the effect of the number of layers of cumulus cells surrounding the oocyte during vitrification of immature equine oocytes. Experimental in vitro and in vivo trials. Immature equine oocytes were vitrified after a short exposure to high concentrations of cryoprotective agents (CPAs), or a long exposure to lower concentrations of CPAs. In Experiment 1, the maturation of oocytes surrounded by multiple layers of cumulus cells (CC oocytes) and oocytes surrounded by only corona radiata (CR oocytes) was investigated. In Experiment 2, spindle configuration was determined for CR oocytes vitrified using the two vitrification protocols. In Experiment 3, further embryonic development was studied after fertilisation and culture. Embryo transfer was performed in a standard manner. Similar nuclear maturation rates were observed for CR oocytes vitrified using the long exposure and nonvitrified controls. Furthermore, a lower maturation rate was obtained for CC oocytes vitrified with the short exposure compared to control CR oocytes (P = 0.001). Both vitrification protocols resulted in significantly higher rates of aberrant spindle configuration than the control groups (Pfoal. The relatively low number of equine oocytes and embryo transfer procedures performed. For vitrification of immature equine oocytes, the use of 1) CR oocytes, 2) a high concentration of CPAs, and 3) a short exposure time may be key factors for maintaining developmental competence. © 2017 EVJ Ltd.

  6. Initiating the Validation of CCIM Processability for Multi-phase all Ceramic (SYNROC) HLW Form: Plan for Test BFY14CCIM-C

    Energy Technology Data Exchange (ETDEWEB)

    Maio, Vince [Idaho National Lab. (INL), Idaho Falls, ID (United States)

    2014-08-01

    This plan covers test BFY14CCIM-C which will be a first–of–its-kind demonstration for the complete non-radioactive surrogate production of multi-phase ceramic (SYNROC) High Level Waste Forms (HLW) using Cold Crucible Induction Melting (CCIM) Technology. The test will occur in the Idaho National Laboratory’s (INL) CCIM Pilot Plant and is tentatively scheduled for the week of September 15, 2014. The purpose of the test is to begin collecting qualitative data for validating the ceramic HLW form processability advantages using CCIM technology- as opposed to existing ceramic–lined Joule Heated Melters (JHM) currently producing BSG HLW forms. The major objectives of BFY14CCIM-C are to complete crystalline melt initiation with a new joule-heated resistive starter ring, sustain inductive melting at temperatures between 1600 to 1700°C for two different relatively high conductive materials representative of the SYNROC ceramic formation inclusive of a HLW surrogate, complete melter tapping and pouring of molten ceramic material in to a preheated 4 inch graphite canister and a similar canister at room temperature. Other goals include assessing the performance of a new crucible specially designed to accommodate the tapping and pouring of pure crystalline forms in contrast to less recalcitrant amorphous glass, assessing the overall operational effectiveness of melt initiation using a resistive starter ring with a dedicated power source, and observing the tapped molten flow and subsequent relatively quick crystallization behavior in pans with areas identical to standard HLW disposal canisters. Surrogate waste compositions with ceramic SYNROC forming additives and their measured properties for inductive melting, testing parameters, pre-test conditions and modifications, data collection requirements, and sampling/post-demonstration analysis requirements for the produced forms are provided and defined.

  7. Thermo-hydro-mechanical processes in the nearfield around a HLW repository in argillaceous formations. Vol. I. Laboratory investigations

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Chun-Liang; Czaikowski, Oliver; Rothfuchs, Tilmann; Wieczorek, Klaus

    2013-06-15

    All over the world, clay formations are being investigated as host medium for geologic disposal of radioactive waste because of their favourable properties, such as very low hydraulic conductivity against fluid transport, good sorption capacity for retardation of radionuclides, and high potential of self-sealing of fractures. The construction of a repository, the disposal of heat-emitting high-level radioactive waste (HLW), the backfilling and sealing of the remaining voids, however, will inevitably induce mechanical (M), hydraulic (H), thermal (T) and chemical (C) disturbances to the host formation and the engineered barrier system (EBS) over very long periods of time during the operation and post-closure phases of the repository. The responses and resulting property changes of the clay host rock and engineered barriers are to be well understood, characterized, and predicted for assessing the long-term performance and safety of the repository.

  8. Comparison of sodium zirconium phosphate-structured HLW forms and synroc for high-level nuclear waste immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Zyryanov, V.N. [Argonne National Lab., IL (United States); Vance, E.R. [ANSTO, Menai (Australia). Materials Division

    1996-12-31

    The incorporation of (a) Cs/Sr as simulated heat-generating isotopes contained in Purex reprocessing waste, (b) simulated actinides, and (c) simulated Purex waste in sodium zirconium phosphate (NZP) has been studied. The samples were prepared by sintering, by hot pressing and by hot isostatic pressing in metal bellows containers. The short-term chemical durability of the phosphate-based material containing Purex waste was within an order of magnitude of that for Synroc-C, as measured by 7-day MCC-1 tests at 90{degrees}C. The dissolution behavior showed evidence of re-precipitation phenomena, even after times as short as 28 days. Potential for improvement of NZP-based ceramics for HLW management is discussed. 19 refs., 4 figs., 3 tabs.

  9. Advances in Glass Formulations for Hanford High-Alumimum, High-Iron and Enhanced Sulphate Management in HLW Streams - 13000

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.

    2013-01-16

    The current estimates and glass formulation efforts have been conservative in terms of achievable waste loadings. These formulations have been specified to ensure that the glasses are homogenous, contain essentially no crystalline phases, are processable in joule-heated, ceramic-lined melters and meet Hanford Tank Waste Treatment and Immobilization Plant (WTP) Contract terms. The WTP?s overall mission will require the immobilization of tank waste compositions that are dominated by mixtures of aluminum (Al), chromium (Cr), bismuth (Bi), iron (Fe), phosphorous (P), zirconium (Zr), and sulphur (S) compounds as waste-limiting components. Glass compositions for these waste mixtures have been developed based upon previous experience and current glass property models. Recently, DOE has initiated a testing program to develop and characterize HLW glasses with higher waste loadings and higher throughput efficiencies. Results of this work have demonstrated the feasibility of increases in waste loading from about 25 wt% to 33-50 wt% (based on oxide loading) in the glass depending on the waste stream. In view of the importance of aluminum limited waste streams at Hanford (and also Savannah River), the ability to achieve high waste loadings without adversely impacting melt rates has the potential for enormous cost savings from reductions in canister count and the potential for schedule acceleration. Consequently, the potential return on the investment made in the development of these enhancements is extremely favorable. Glass composition development for one of the latest Hanford HLW projected compositions with sulphate concentrations high enough to limit waste loading have been successfully tested and show tolerance for previously unreported tolerance for sulphate. Though a significant increase in waste loading for high-iron wastes has been achieved, the magnitude of the increase is not as substantial as those achieved for high-aluminum, high-chromium, high-bismuth or sulphur

  10. Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

    OpenAIRE

    Gómez, E.; Rodríguez, A; Muñoz, M.; Caamaño, J.N. (José); Hidalgo, C.O. (Carlos); Morán, E.; Facal, Nieves; Díez, C.

    2013-01-01

    The aim of this study was to examine the effects of co-culture with Vero cells during the in vitro maturation (IVM) and three culture media, B2+5% fetal calf serum (FCS) on Vero cells, synthetic oviduct fluid (SOF)+5% FCS, and SOF+20 gL(-1) bovine serum albumin (BSA), on the developmental competence of the embryos and their ability to survive vitrification/warming. We also tested the effect of morphological quality and the age of the embryo on its sensitivity to vitrification. The IVM system ...

  11. Technology evaluation report: Babcock and Wilcox Cyclone Furnace Vitrification technology. Volume 1

    Energy Technology Data Exchange (ETDEWEB)

    Groeber, P.

    1992-09-01

    The project consists of an analysis of the Babcock and Wilcox (B and W) Cyclone Furnace Vitrification process. The SITE Demonstration took place at the B and W Research and Development Division in Alliance, Ohio. The vitrification process was performed on a synthetic soil matrix (SSM) that was spiked with known concentrations of semivolatile organic compounds, metals, and simulated radionuclides. The Demonstration effort was directed at obtaining information on the performance and cost of the process for use at other sites. Documentation will consist of two reports. This Technology Evaluation Report (TER) is contained in two volumes and describes the field activities and laboratory results.

  12. Vitrification of cesium-contaminated organic ion exchange resin

    Energy Technology Data Exchange (ETDEWEB)

    Sargent, Jr., Thomas N. [Clemson Univ., SC (United States)

    1994-08-01

    Vitrification has been declared by the Environmental Protection Agency (USEPA) as the Best Demonstrated Available Technology (BDAT) for the permanent disposal of high-level radioactive waste. Savannah River Site currently uses a sodium tetraphenylborate (NaTPB) precipitation process to remove Cs-137 from a wastewater solution created from the processing of nuclear fuel. This process has several disadvantages such as the formation of a benzene waste stream. It has been proposed to replace the precipitation process with an ion exchange process using a new resorcinol-formaldehyde resin developed by Savannah River Technical Center (SRTC). Preliminary tests, however, showed that problems such as crust formation and a reduced final glass wasteform exist when the resin is placed in the melter environment. The newly developed stirred melter could be capable of overcoming these problems. This research explored the operational feasibility of using the stirred tank melter to vitrify an organic ion exchange resin. Preliminary tests included crucible studies to determine the reducing potential of the resin and the extent of oxygen consuming reactions and oxygen transfer tests to approximate the extent of oxygen transfer into the molten glass using an impeller and a combination of the impeller and an external oxygen transfer system. These preliminary studies were used as a basis for the final test which was using the stirred tank melter to vitrify nonradioactive cesium loaded organic ion exchange resin. Results from this test included a cesium mass balance, a characterization of the semi-volatile organic compounds present in the off gas as products of incomplete combustion (PIC), a qualitative analysis of other volatile metals, and observations relating to the effect the resin had on the final redox state of the glass.

  13. Flammability Control In A Nuclear Waste Vitrification System

    Energy Technology Data Exchange (ETDEWEB)

    Zamecnik, John R.; Choi, Alexander S.; Johnson, Fabienne C.; Miller, Donald H.; Lambert, Daniel P.; Stone, Michael E.; Daniel, William E. Jr.

    2013-07-25

    The Defense Waste Processing Facility at the Savannah River Site processes high-level radioactive waste from the processing of nuclear materials that contains dissolved and precipitated metals and radionuclides. Vitrification of this waste into borosilicate glass for ultimate disposal at a geologic repository involves chemically modifying the waste to make it compatible with the glass melter system. Pretreatment steps include removal of excess aluminum by dissolution and washing, and processing with formic and nitric acids to: 1) adjust the reduction-oxidation (redox) potential in the glass melter to reduce radionuclide volatility and improve melt rate; 2) adjust feed rheology; and 3) reduce by steam stripping the amount of mercury that must be processed in the melter. Elimination of formic acid in pretreatment has been studied to eliminate the production of hydrogen in the pretreatment systems, which requires nuclear grade monitoring equipment. An alternative reductant, glycolic acid, has been studied as a substitute for formic acid. However, in the melter, the potential for greater formation of flammable gases exists with glycolic acid. Melter flammability is difficult to control because flammable mixtures can be formed during surges in offgases that both increase the amount of flammable species and decrease the temperature in the vapor space of the melter. A flammable surge can exceed the 60% of the LFL with no way to mitigate it. Therefore, careful control of the melter feed composition based on scaled melter surge testing is required. The results of engineering scale melter tests with the formic-nitric flowsheet and the use of these data in the melter flammability model are presented.

  14. The Conceptual Mechanism for Viable Organizational Learning Based on Complex System Theory and the Viable System Model

    Science.gov (United States)

    Sung, Dia; You, Yeongmahn; Song, Ji Hoon

    2008-01-01

    The purpose of this research is to explore the possibility of viable learning organizations based on identifying viable organizational learning mechanisms. Two theoretical foundations, complex system theory and viable system theory, have been integrated to provide the rationale for building the sustainable organizational learning mechanism. The…

  15. Deletion of ultraconserved elements yields viable mice

    Energy Technology Data Exchange (ETDEWEB)

    Ahituv, Nadav; Zhu, Yiwen; Visel, Axel; Holt, Amy; Afzal, Veena; Pennacchio, Len A.; Rubin, Edward M.

    2007-07-15

    Ultraconserved elements have been suggested to retainextended perfect sequence identity between the human, mouse, and ratgenomes due to essential functional properties. To investigate thenecessities of these elements in vivo, we removed four non-codingultraconserved elements (ranging in length from 222 to 731 base pairs)from the mouse genome. To maximize the likelihood of observing aphenotype, we chose to delete elements that function as enhancers in amouse transgenic assay and that are near genes that exhibit markedphenotypes both when completely inactivated in the mouse as well as whentheir expression is altered due to other genomic modifications.Remarkably, all four resulting lines of mice lacking these ultraconservedelements were viable and fertile, and failed to reveal any criticalabnormalities when assayed for a variety of phenotypes including growth,longevity, pathology and metabolism. In addition more targeted screens,informed by the abnormalities observed in mice where genes in proximityto the investigated elements had been altered, also failed to revealnotable abnormalities. These results, while not inclusive of all thepossible phenotypic impact of the deleted sequences, indicate thatextreme sequence constraint does not necessarily reflect crucialfunctions required for viability.

  16. Is Greenberg's "Macro-Carib" viable?

    Directory of Open Access Journals (Sweden)

    Spike Gildea

    Full Text Available In his landmark work Language in the Americas, Greenberg (1987 proposed that Macro-Carib was one of the major low-level stocks of South America, which together with Macro-Panoan and Macro-Ge-Bororo were claimed to comprise the putative Ge-Pano-Carib Phylum. His Macro-Carib includes the isolates Andoke and Kukura, and the Witotoan, Peba-Yaguan, and Cariban families. Greenberg's primary evidence came from person-marking paradigms in individual languages, plus scattered words from individual languages collected into 79 Macro-Carib 'etymologies' and another 64 Amerind 'etymologies'. The goal of this paper is to re-evaluate Greenberg's Macro-Carib claim in the light of the much more extensive and reliable language data that has become available largely since 1987. Based on full person-marking paradigms for Proto-Cariban, Yagua, Bora and Andoke, we conclude that Greenberg's morphological claims are unfounded. For our lexical comparison, we created lexical lists for Proto-Cariban, Proto-Witotoan, Yagua and Andoke, for both Greenberg's 143 putative etymologies and for the Swadesh 100 list. From both lists, a total of 23 potential cognates were found, but no consonantal correspondences were repeated even once. We conclude that our greatly expanded and improved database does not provide sufficient evidence to convince the skeptic that the Macro-Carib hypothesis is viable

  17. Economically viable large-scale hydrogen liquefaction

    Science.gov (United States)

    Cardella, U.; Decker, L.; Klein, H.

    2017-02-01

    The liquid hydrogen demand, particularly driven by clean energy applications, will rise in the near future. As industrial large scale liquefiers will play a major role within the hydrogen supply chain, production capacity will have to increase by a multiple of today’s typical sizes. The main goal is to reduce the total cost of ownership for these plants by increasing energy efficiency with innovative and simple process designs, optimized in capital expenditure. New concepts must ensure a manageable plant complexity and flexible operability. In the phase of process development and selection, a dimensioning of key equipment for large scale liquefiers, such as turbines and compressors as well as heat exchangers, must be performed iteratively to ensure technological feasibility and maturity. Further critical aspects related to hydrogen liquefaction, e.g. fluid properties, ortho-para hydrogen conversion, and coldbox configuration, must be analysed in detail. This paper provides an overview on the approach, challenges and preliminary results in the development of efficient as well as economically viable concepts for large-scale hydrogen liquefaction.

  18. Vitrification of radioactive waste. Application to other kinds of waste; Vitrification des dechets radioactifs. Application a d`autres types de dechets

    Energy Technology Data Exchange (ETDEWEB)

    Jouan, A.

    1993-12-31

    The containment by vitrification of radioactive waste is applied to concentrate solutions of fission products coming from the spent fuel reprocessing. By the way of liquid state to solid state, it is possible to reduce the volume of waste, to get a material with safety guarantees necessary to long storage and the glass by its chemical resistance, its thermal stability and its well resistance to irradiation answers particularly well to these necessities.

  19. Comparison of the level(s) of DNA damage using Comet assay in bovine oocytes subjected to selected vitrification methods.

    Science.gov (United States)

    Stachowiak, E M; Papis, K; Kruszewski, M; Iwaneńko, T; Bartłomiejczyk, T; Modliński, J A

    2009-08-01

    It was suggested that the cryodamage to oocytes' DNA has been responsible for the compromised developmental competence of cryopreserved oocytes. Vitrification of bovine oocytes affected not only cellular components, but also nuclear material. A significant rate of DNA fragmentation was found in bovine frozen or vitrified oocytes analysed by Comet assay regardless of cryopreservation method. Our method of vitrification using droplet system after gentle pre-equilibration treatment is one of the most effective cryopreservation methods employed for bovine oocytes so far, making it possible to develop 30% blastocyst stage embryos. In this study, the extent of DNA damage in bovine oocytes vitrified using three vitrification methods (droplet system, Open Pulled Straw and traditional vitrification in 0.25 ml insemination straws) was compared using Comet assay. Vitrification in droplet system and Open Pull Straws vitrification did not result in detectable cryoinjuries of DNA of bovine oocytes. On the contrary, DNA fragmentation was found in four of 26 oocytes vitrified in 0.25 ml straws (15.4%, p vitrification methods).

  20. Parejas viables que perduran en el tiempo

    Directory of Open Access Journals (Sweden)

    Juan José Cuervo Rodríguez

    2013-01-01

    Full Text Available El presente artículo científico presenta resultados del proceso llevado a cabo en el proyecto de investigación docente "Mecanismos de autorregulación en parejas viables que perduran en el tiempo". Se soporta en una mirada compleja de la psicología basada en una epistemología de la construcción. En el ámbito metodológico, se inscribe en los estudios de terapia familiar desde una perspectiva de la comunicación humana como un todo integrado. Participaron nueve parejas. Los criterios de inclusión fueron: cinco o más años de convivencia, participación voluntaria, no presentar (ni haber presentado problemáticas especiales que ameriten intervención psicoterapéutica y la obtención de un porcentaje significativo en el uso de estrategias de comunicación asertiva en la resolución de conflictos. El método general utilizado fue el análisis de la comunicación en tarea de conversación. Los principales hallazgos señalan una estrecha relación entre el contexto de desarrollo de las parejas, la emergencia de códigos comunicacionales propios y la posibilidad de perdurar en el tiempo; también, se resalta el tipo de comunicación asertiva o constructiva, la construcción de valores como el respeto y la aceptación de las diferencias, y el deseo por vivir y construir bienestar común, como elementos constitutivos de su identidad como pareja.

  1. Pretreatment of in vitro matured bovine oocytes with docetaxel before vitrification: Effects on cytoskeleton integrity and developmental ability after warming.

    Science.gov (United States)

    Chasombat, Jakkhaphan; Nagai, Takashi; Parnpai, Rangsun; Vongpralub, Thevin

    2015-10-01

    The stabilization of spindle fibersis important for successful vitrification of bovine oocytes because microtubules and other cytoskeleton fibers (CSF) can be damaged during vitrification, resulting in failure of fertilization after thawing. Docetaxel, a stabilizing agent, could potentially reduce CSF damage of bovine oocytes induced during vitrification. However, there have been no reports on the effects of docetaxel on their vitrification. Experiment 1 was conducted to investigate the effects of various doses of docetaxel (0.0, 0.05, 0.5, 5.0 and 50 μM) in preincubation medium of in vitro matured (IVM) bovine oocytes on their developmental ability after in vitro fertilization (IVF). The results show that 0.05 μM docetaxel had no adverse effect on embryo development, while docetaxel at a concentration of ⩾0.5 μM inhibited development. Experiments 2 and 3 were conducted to investigate the effects of preincubation of IVM bovine oocytes with 0.05 μM docetaxel for 30 min prior to vitrification-warming on CSF integrity (Experiment 2), and on oocyte survival and viability after IVF (Experiment 3). When preincubated with 0.05 μM docetaxel for 30 min before vitrification, post-thawed oocytes had less CSF damage and higher survival rates compared with those untreated with docetaxel before vitrification. Surviving oocytes also had higher rates of cleavage and development to the blastocyst stage after IVF. In conclusion, preincubation of IVM bovine oocytes with 0.05 μM docetaxel for 30 min before vitrification was effective at preventing CSF damage during vitrification, and improving oocyte viability after warming and subsequent cleavage and blastocyst formation after IVF. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Closed pulled straw vitrification of in vitro-produced and in vivo-produced bovine embryos.

    Science.gov (United States)

    Yu, X L; Deng, W; Liu, F J; Li, Y H; Li, X X; Zhang, Y L; Zan, L S

    2010-03-01

    The objective of this study was to evaluate the efficiency of the closed pulled straw (CPS) method for cryopreserving in vitro-produced and in vivo-produced bovine (Bos taurus) embryos. Based on the open pulled straw (OPS) protocol, the top end of a CPS was closed by tweezers (heated in a flame) to prevent the cryoprotectant medium containing embryos from contacting the liquid nitrogen. Bovine in vitro or in vivo morulae and early blastocyst embryos were frozen by slow cryopreservation, OPS vitrification, or CPS vitrification. Morphology of postthawed embryos was evaluated, and normal embryos were used for successive culture for 72h. There were no significant differences between OPS and CPS freezing groups in postthawed in vitro-produced embryos with respect to rates of morphologically normal embryos (mean+/-SD, 87.9+/-5.2% vs. 85.4+/-4.9%), survival at 24h (58.0+/-6.8% vs. 56.3+/-4.4%), and survival at 72h (35.2+/-6.0% vs. 34.9+/-6.7%). However, both OPS and CPS vitrification resulted in higher postthaw rates of morphologically normal embryo and survival at 24 and 72h than those of the slow-freezing method (Pvitrification was a feasible method to cryopreserve both in vitro-derived and in vivo-derived bovine embryos. This method not only eliminated the risk of embryo contamination by preventing contact with liquid nitrogen but also retained the advantages of the OPS vitrification method. Copyright 2010. Published by Elsevier Inc.

  3. Cholesterol added prior to vitrification on the cryotolerance of immature and in vitro matured bovine oocytes.

    Science.gov (United States)

    Arcarons, Núria; Morató, Roser; Vendrell, Meritxell; Yeste, Marc; López-Bejar, Manel; Rajapaksha, Kosala; Anzar, Muhammad; Mogas, Teresa

    2017-01-01

    This study examines whether incorporating cholesterol-loaded methyl-β-cyclodextrin (CLC) in the bovine oocyte plasma membrane improves oocyte tolerance to vitrification. In vitro matured oocytes were incubated with 2 mg/ml BODIPY-labeled CLC for different time intervals in FCS or PVA supplemented medium or exposed to different CLC concentrations to examine the subcellular localization of cholesterol by confocal microscopy live-cell imaging. Subsequently, the effects of optimized CLC concentrations and incubation times prior to vitrification on early embryo development were assessed. Then, we evaluated the effects of pretreatment with 2 mg/ml CLC for 30 min before the vitrification of immature (GV) and in vitro matured (MII) oocytes on developmental competence and gene expression. Our results indicate a high plasma membrane labeling intensity after 30 min of incubation with 2 mg/ml CLC for 30 min, regardless of the holding medium used. When oocytes were incubated with 1 mg/ml, 2 mg/ml and 3 mg/ml of CLC, intense labeling was observed at the plasma membrane after 40, 30 and 20 min, respectively. CLC pre-treatment before the vitrification of bovine oocytes did not affect subsequent cleavage and embryo development rates irrespective of CLC concentrations, incubation times or meiotic stage. However, pretreatment seems to improve the quality of embryos derived from vitrified oocytes, mainly when oocytes were vitrified at the GV stage.

  4. A chronologic review of mature oocyte vitrification research in cattle, pigs, and sheep.

    Science.gov (United States)

    Mullen, S F; Fahy, G M

    2012-11-01

    Vitrification as a means of cryopreservation has become a standard approach for oocytes from livestock. This paradigm shift occurred primarily as a result of the demonstration in 1996 that bovine oocytes are extremely susceptible to chilling injury. Since that early work, numerous devices have been used as supports for oocytes during so-called "ultra-rapid cooling", and occasionally, trials involving the deposition of small volumes of media containing oocytes directly into liquid nitrogen to facilitate cooling have been reported. Results reporting blastocyst development exceeding 10% are common, but variability remains high, and a standard method for bovine oocytes remains to be established. Oocytes from pigs are particularly difficult to cryopreserve, even with the use of ultrarapid cooling approaches. Few reports have demonstrated blastocyst development exceeding 5%. The application of hydrostatic pressure before vitrification appears to impart stress tolerance to porcine oocytes, as the results of some treatments have shown development to blastocysts at proportions >10%. Work on sheep oocyte vitrification is relatively new, and a few articles have reported blastocyst development at 10% or more. Messenger RNA levels are reportedly altered in sheep oocytes as a result of vitrification, and damage to the cytoskeleton is common across species. Copyright © 2012 Elsevier Inc. All rights reserved.

  5. Cholesterol added prior to vitrification on the cryotolerance of immature and in vitro matured bovine oocytes.

    Directory of Open Access Journals (Sweden)

    Núria Arcarons

    Full Text Available This study examines whether incorporating cholesterol-loaded methyl-β-cyclodextrin (CLC in the bovine oocyte plasma membrane improves oocyte tolerance to vitrification. In vitro matured oocytes were incubated with 2 mg/ml BODIPY-labeled CLC for different time intervals in FCS or PVA supplemented medium or exposed to different CLC concentrations to examine the subcellular localization of cholesterol by confocal microscopy live-cell imaging. Subsequently, the effects of optimized CLC concentrations and incubation times prior to vitrification on early embryo development were assessed. Then, we evaluated the effects of pretreatment with 2 mg/ml CLC for 30 min before the vitrification of immature (GV and in vitro matured (MII oocytes on developmental competence and gene expression. Our results indicate a high plasma membrane labeling intensity after 30 min of incubation with 2 mg/ml CLC for 30 min, regardless of the holding medium used. When oocytes were incubated with 1 mg/ml, 2 mg/ml and 3 mg/ml of CLC, intense labeling was observed at the plasma membrane after 40, 30 and 20 min, respectively. CLC pre-treatment before the vitrification of bovine oocytes did not affect subsequent cleavage and embryo development rates irrespective of CLC concentrations, incubation times or meiotic stage. However, pretreatment seems to improve the quality of embryos derived from vitrified oocytes, mainly when oocytes were vitrified at the GV stage.

  6. Role of cumulus cells during vitrification and fertilization of mature bovine oocytes

    NARCIS (Netherlands)

    Ortiz-Escribano, N.; Smits, K.; Piepers, S.; Abbeel, Van den E.; Woelders, H.; Soom, Van A.

    2016-01-01

    This study was designed to determine the role of cumulus cells during vitrification of bovine oocytes. Mature cumulus-oocyte complexes (COCs) with many layers of cumulus cells, corona radiata oocytes (CRs), with a few layers of cumulus cells, and denuded oocytes (DOs) without cumulus cells were

  7. Vitrification of in vitro produced bovine blastocysts: methodological studies and developmental capacity.

    Science.gov (United States)

    Donnay, I; Auquier, P; Kaidi, S; Carolan, C; Lonergan, P; Mermillod, P; Massip, A

    1998-08-21

    Methodological studies were undertaken to test the validity of a three-step vitrification procedure for bovine in vitro produced embryos using glycerol and ethylene glycol as cryoprotectants. Embryos were produced in a low-phosphate culture system (medium VT1 + 10% foetal calf serum) and vitrified at day 7 post-insemination either in a mixture of 25% glycerol--25% ethylene glycol or a mixture of 10% glycerol--40% ethylene glycol. In the first mixture 67% (n = 283) of blastocysts were re-expanded after 72 h of culture and 53% were hatched while in the second one (n = 65) only 5% survived. The mean number of cells of the surviving blastocysts was correlated with the rate of survival (R2 = 0.47; P = 0.0024). Embryo size (diameter to 180 microm) did not influence blastocyst survival or cell number, but hatching rate was higher for embryos > 180 microm. Embryo survival, hatching rate and cell number 72 h post-warming were not affected by the mode of vitrification (direct plunging into nitrogen liquid or vitrification into nitrogen liquid vapour), the mode of preparation of the vitrification solutions (molar or molal basis) or by the concentration of galactose used as a diluent (0 to 0.85 M). Only one calf was born after transfer of 22 vitrified blastocysts. These results confirm the apparent lack of correlation for cryopreserved embryos between in vitro survival or hatching and viability after transfer.

  8. Ultrastructure of bovine in vitro-produced blastocysts cryopreserved by vitrification.

    Science.gov (United States)

    Ohboshi, S; Fujihara, N; Yoshida, T; Tomagane, H

    1998-02-01

    The objective of this study was to examine ultrastructural aspects of bovine in vitro-produced blastocysts associated with cryopreservation by vitrification. Morphologically good embryos were used and treated with ethylene-glycol-based vitrification solution (VS). The untreated embryos had conventional fine structure. The post-warming embryos treated with direct exposure to VS (one-step procedure) showed cellular damage structurally by cryopreservation, which included loss of microvilli, disruption of the plasma membrane, mitochondrial changes and swelling of the endoplasmic reticulum. However, nuclei and junctional regions seemed to be resistant to cryoinjury. In contrast, the post-warming embryos pre-equilibrated with 10% ethylene glycol for 5 min and subsequent exposure to VS (two-step procedure) showed less damage than those treated by the one-step procedure. Post-warming embryos treated by the two-step procedure were cultured in vitro for 18 h. Some embryos survived and their structures re-formed to the former state, while other embryos showed serious injuries and could not reconstitute the blastocoele. Three post-warming embryos treated by the two-step procedure that survived after in vitro culture were transferred to three recipients and one of these resulted in pregnancy. These results indicate that cryopreservation by vitrification can damage membranous structures of the cells of bovine embryos, the extent and nature of this damage being dependent on the vitrification procedure.

  9. Bovine oocyte vitrification before or after meiotic arrest: effects on ultrastructure and developmental ability.

    Science.gov (United States)

    Diez, Carmen; Duque, Paloma; Gómez, Enrique; Hidalgo, Carlos O; Tamargo, Carolina; Rodríguez, Aida; Fernández, Lina; de la Varga, Santiago; Fernández, Alba; Facal, Nieves; Carbajo, Maite

    2005-07-15

    The nuclear stage at which oocytes are cryopreserved influences further development ability and cryopreservation affects ultrastructure of both cumulus cells and the oocyte. In this work, we analyze the effects of vitrification at different nuclear and cytoplasmic maturation stages on the oocyte ultrastructure and developmental ability. Culture in TCM199 + PVA with roscovitine 25 M during 24h led to meiotic arrest (MA) in cumulus-oocyte complexes (COCs), while permissive in vitro maturation (IVM) was performed in TCM199, 10% FCS, FSH-LH and 17beta-estradiol for 24 h. Oocytes were vitrified using the open pulled straw method (OPS) with minor modifications. Fresh and vitrified/warmed COCs were fixed as immature, after IVM, after meiotic arrest (MA) and after MA + IVM. Vitrification combined with MA followed by IVM produced the highest rates of degeneration, regardless of the vitrification time. As a consequence, lower proportions of embryos cleaved in these groups, although differences were eliminated at the five-eight cell stage. Development rates up to day 8 were similar in all experimental groups, being significantly lower than those in fresh controls. Only oocytes vitrified after IVM were able to give blastociysts. The morphological alterations observed can be responsible for compromised development. More research is needed to explain the low survival rates of the bovine oocyte after vitrification and warming.

  10. 76 FR 13605 - Notice of Availability of Draft Waste Incidental to Reprocessing Evaluation for the Vitrification...

    Science.gov (United States)

    2011-03-14

    ... vitrified the waste (combined it at a high temperature with borosilicate glass) and transferred the molten glass-waste mixture into specially developed ] stainless steel canisters where the mixture hardened into a solid glass waste form. DOE used the vitrification melter as part of this process, specifically to...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2014-10-01

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

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

    Science.gov (United States)

    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...

  14. Effect of NaOH on the vitrification process of waste Ni-Cr sludge

    Energy Technology Data Exchange (ETDEWEB)

    Chou, I-Cheng [Department of Safety Health and Environmental Engineering, Chung Hwa University of Medical Technology, 89 Wenhwa 1st St., Rende Shiang, Tainan County 71703, Taiwan (China); Wang, Ya-Fen [Department of Bioenvironmental Engineering and R and D Center of Membrane Technology, Chung Yuan Christian University, Chung-Li 320, Taiwan (China); Chang, Cheng-Ping [Institute of Occupational Safety and Health, Council of Labor Affairs, Taipei City, Taiwan (China); Wang, Chih-Ta [Department of Safety Health and Environmental Engineering, Chung Hwa University of Medical Technology, 89 Wenhwa 1st St., Rende Shiang, Tainan County 71703, Taiwan (China); Kuo, Yi-Ming, E-mail: yiming@mail.hwai.edu.tw [Department of Safety Health and Environmental Engineering, Chung Hwa University of Medical Technology, 89 Wenhwa 1st St., Rende Shiang, Tainan County 71703, Taiwan (China)

    2011-01-30

    This study investigated the effect of NaOH on the vitrification of electroplating sludge. Ni, the major metal in the electroplating sludge, is the target for recovery in the vitrification. Sludge and encapsulation materials (dolomite, limestone, and cullet) were mixed and various amounts of NaOH were added to serve as a glass modifier and a flux. A vitrification process at 1450 deg. C separated the molten specimens into slag and ingot. The composition, crystalline characteristics, and leaching characteristics of samples were measured. The results indicate that the recovery of Ni is optimal with a 10% NaOH mass ratio; the recoveries of Fe, Cr, Zn, Cu, and Mn all exhibited similar trends. The results of the toxicity characteristic leaching procedure (TCLP) show that leaching characteristics of the slag meet the requirements of regulation in Taiwan. In addition, a semi-quantitative X-ray diffraction analysis revealed that the main crystalline phase of slag changed from Ca{sub 3}(Si{sub 3}O{sub 9}) to Na{sub 4}Ca{sub 4}(Si{sub 6}O{sub 18}) with a NaOH mass ratio of over 15%, because the Ca{sup 2+} ions were replaced with Na{sup +} ions during the vitrification process. Na{sub 4}Ca{sub 4}(Si{sub 6}O{sub 18}), a complex mineral which hinders the mobility of metals, accounts for the decrease of metal recovery.

  15. BULK VITRIFICATION TECHNOLOGY FOR THE TREATMENT AND IMMOBILIZATION OF LOW-ACTIVITY WASTE

    Energy Technology Data Exchange (ETDEWEB)

    ARD KE

    2011-04-11

    This report is one of four reports written to provide background information regarding immobilization technologies under consideration for supplemental immobilization of Hanford's low-activity waste. This paper is intended to provide the reader with general understanding of Bulk Vitrification and how it might be applied to immobilization of Hanford's low-activity waste.

  16. Effect of vitrification and post-thawing interval on the cytoskeleton ...

    African Journals Online (AJOL)

    Lucky Nedambale

    2,3# ... Keywords: Vitrification, cytoskeleton, bovine, oocytes, fertilization, in vitro matured. # Corresponding author. ... the sperm for 6 h in 5% CO2 in air at 39 °C. After 6 h of IVF, oocytes were washed six times in TL-HEPES. (Bio-Whittaker ...

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

    Science.gov (United States)

    Petunia (Petunia × hybrida Vilm.) is a very important crop conserved in the National Genebank of China. Petunia cultivar “Niu 2” was used to develop a droplet-vitrification protocol to cryopreserve shoot tips. Six variables (age of the in vitro plants, concentration of sucrose in the preculture solu...

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

    NARCIS (Netherlands)

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

    2013-01-01

    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

  19. Vitrification of human immature oocytes before and after in vitro maturation: a review.

    Science.gov (United States)

    Khalili, Mohammad Ali; Shahedi, Abbas; Ashourzadeh, Sareh; Nottola, Stefania Annarita; Macchiarelli, Guido; Palmerini, Maria Grazia

    2017-08-18

    The use of immature oocytes subjected to in vitro maturation (IVM) opens interesting perspectives for fertility preservation where ovarian reserves are damaged by pathologies or therapies, as in PCO/PCOS and cancer patients. Human oocyte cryopreservation may offer some advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation and postponing childbirth. It also eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In addition, a successful oocyte cryopreservation program could eliminate the need for donor and recipient menstrual cycle synchronization. Recent advances in vitrification technology have markedly improved the oocyte survival rate after warming, with fertilization and implantation rates comparable with those of fresh oocytes. Healthy live births can be achieved from the combination of IVM and vitrification, even if vitrification of in vivo matured oocytes is still more effective. Recently, attention is given to highlight whether vitrification procedures are more successful when performed before or after IVM, on immature GV-stage oocytes, or on in vitro matured MII-stage oocytes. In this review, we emphasize that, even if there are no differences in survival rates between oocytes vitrified prior to or post-IVM, reduced maturation rates of immature oocytes vitrified prior to IVM can be, at least in part, explained by underlying ultrastructural and biomolecular alterations.

  20. Obesity does not aggravate vitrification injury in mouse embryos: a prospective study.

    Science.gov (United States)

    Ma, Wenhong; Yang, Xing; Liang, Xiaoyan

    2012-08-31

    Obesity is associated with poor reproductive outcomes, but few reports have examined thawed embryo transfer in obese women. Many studies have shown that increased lipid accumulation aggravates vitrification injury in porcine and bovine embryos, but oocytes of these species have high lipid contents (63 ng and 161 ng, respectively). Almost nothing is known about lipids in human oocytes except that these cells are anecdotally known to be relatively lipid poor. In this regard, human oocytes are considered to be similar to those of the mouse, which contain approximately 4 ng total lipids/oocyte. To date, no available data show the impact of obesity on vitrification in mouse embryos. The aim of this study was to establish a murine model of maternal diet-induced obesity and to characterize the effect of obesity on vitrification by investigating the survival rate and embryo developmental competence after thawing. Prospective comparisons were performed between six-eight-cell embryos from obese and normal-weight mice and between fresh and vitrified embryos. Female C57BL/6 mice were fed standard rodent chow (normal-weight group) or a high-fat diet (obese group) for 6 weeks. The mice were mated, zygotes were collected from oviducts and cultured for 3 days, and six-eight-cell embryos were then selected to assess lipid content in fresh embryos and to evaluate differences in apoptosis, survival, and development rates in response to vitrification. In fresh embryos from obese mice, the lipid content (0.044 vs 0.030, Pvitrification, no significant difference was found between thawed embryos from obese and normal-weight mice in apoptosis, survival, and development rates on days 4 and 5. In both groups, pre- and post-vitrification embryo apoptosis, survival, and development rates were similar. This study demonstrated that differences in survival and developmental rates between embryos from obese and normal-weight mice were eliminated after vitrification. Thus, maternal obesity does

  1. An efficient method for the sanitary vitrification of bovine oocytes in straws.

    Science.gov (United States)

    Zhou, Yanhua; Fu, Xiangwei; Zhou, Guangbin; Jia, Baoyu; Fang, Yi; Hou, Yunpeng; Zhu, Shien

    2014-01-01

    At present, vitrification has been widely applied to humans, mice and farm animals. To improve the efficiency of vitrification in straw, bovine oocytes were used to test a new two-step vitrification method in this study. When in vitro matured oocytes were exposed to 20% ethylene glycol (EG20) for 5 min and 40% ethylene glycol (EG40) for 30 s, followed by treatment with 30% glycerol (Gly30), Gly40 or Gly50, a volume expansion was observed in Gly30 and Gly40 but not Gly50. This indicates that the intracellular osmotic pressure after a 30 s differs between EG40 and ranged between Gly40 (approximately 5.6 mol/L) and Gly50 (approximately 7.0 mol/L). Since oocytes are in EG40 just for only a short period of time (30 s) and at a lower temperature (4°C), we hypothesize that the main function of this step in to induce dehydration. Based on these results, we omitted the EG40 step, before oocytes were pretreated in EG20 for 5 min, exposed to pre-cooled (4°C) Gly50, for 30 s, and then dipped into liquid nitrogen. After warming, 81.1% of the oocytes survived, and the surviving oocytes developed into cleavage stage embryos (63.5%) or blastocysts (20.0%) after parthenogenetic activation. These results demonstrate that in a two-step vitrification procedure, the permeability effect in the second step is not necessary. It is possible that the second step is only required to provide adequate osmotic pressure to condense the intracellular concentration of CPAs to a level required for successful vitrification.

  2. Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification.

    Science.gov (United States)

    Gómez, E; Rodríguez, A; Muñoz, M; Caamaño, J N; Hidalgo, C O; Morán, E; Facal, N; Díez, C

    2008-05-01

    The aim of this study was to examine the effects of co-culture with Vero cells during the in vitro maturation (IVM) and three culture media, B2+5% fetal calf serum (FCS) on Vero cells, synthetic oviduct fluid (SOF)+5% FCS, and SOF+20 gL(-1) bovine serum albumin (BSA), on the developmental competence of the embryos and their ability to survive vitrification/warming. We also tested the effect of morphological quality and the age of the embryo on its sensitivity to vitrification. The IVM system neither affects the embryo development up to Day 7 nor survival rates after vitrification. The culture of embryos in SOF+FCS and in Vero cells+B2 allowed obtaining more Day 6 and Day 7 blastocysts, and a higher % of Day 7 blastocysts vitrified than culture in SOF+BSA. Contrarily, on Day 8, more blastocysts were vitrified in SOF+BSA than in SOF+FCS. Blastocysts quality affected development after vitrification/warming, and Day 7 embryos showed higher survival rates than their Day 8 counterparts. Day 7 blastocysts produced in Vero cells or in SOF+BSA survived at higher rates than those produced in SOF+FCS at 24 and 48 h after warming. Embryo culture with BSA allows obtaining hatching rates after vitrification/warming higher than those obtained after co-culture with Vero cells in B2 and FCS. Moreover, this system provides hatching rates from Day 8 blastocysts comparable to those obtained on Day 7 in Vero cells. Further studies, including embryo transfer to recipients, are needed to clarify factors affecting the freezability of in vitro produced bovine embryos.

  3. Vitrification by Cryotop and the Maturation, Fertilization, and Developmental Rates of Mouse Oocytes

    Science.gov (United States)

    Abedpour, Neda; Rajaei, Farzad

    2015-01-01

    Background: Oocyte cryopreservation is an important part of modern fertility treatment. The effect of vitrification on the fertilization and developmental rates of embryo is still a matter of debate. Objectives: This study aimed to investigate the effect of vitrification on the success of mouse oocyte maturation, fertilization, and preimplantation development in vitro. Materials and Methods: In this experimental study, a total of 200 germinal vesicle (GV) and 200 metaphase II (MII) oocytes were obtained from ovaries and fallopian tubes of NMRI mice, respectively and divided into two control and experimental (vitrified) groups. Oocytes in the experimental group were vitrified by Cryotop using vitrification medium (Origio, Denmark) and kept in liquid nitrogen for one month. Then, they were cultured in maturation medium for 24 hours. In vitro maturated metaphase 2 (IVM-MII) and ovulated metaphase 2 (OV-MII) oocytes were inseminated and the fertilized embryos assessed until the hatching blastocyst stage. Outcomes were assessed for statistical significance by Chi-square test using SPSS software. Results: Vitrification caused a significant reduction in the maturation rate of oocytes. Of those that matured, the fertilization rate of vitrified IVM-MII (44.1%) and OV-MII oocytes (50%) was not significantly different from each other but both were significantly lower than the control group (P < 0.05). There was no significant difference in developmental rates of both vitrified groups and the control group. Conclusions: The present study showed that vitrification using Cryotop and freezing medium can damage oocytes by reducing the maturation and fertilization rates in both developmental stages. PMID:26568845

  4. Impact of multiple blastocyst biopsy and vitrification-warming procedures on pregnancy outcomes.

    Science.gov (United States)

    Bradley, Cara K; Livingstone, Mark; Traversa, Maria V; McArthur, Steven J

    2017-12-01

    To assess the impact of multiple blastocyst biopsy and vitrification-warming procedures on clinical outcomes. Retrospective study. Private fertility clinic. Preimplantation genetic diagnosis (PGD) patients undergoing comprehensive chromosome screening, including monogenic disorder and chromosome rearrangement cases. Warming and transfer of euploid blastocysts biopsied and vitrified-warmed once (group 1 [G1, control]; n = 2,130), biopsied once but vitrified-warmed twice (group 2 [G2]; n = 34), or biopsied and vitrified-warmed twice (group 3 [G3]; n = 29). Thaw (for transfer) survival rate and clinical pregnancy rate (CPR). The thaw survival rates were 98.4% for G1, 97.3% for G2, and 93.3% for G3, with once biopsied and vitrified-warmed embryos being significantly higher than twice biopsied and vitrified-warmed embryos (G1 vs. G3; P=.032). There was a slight reduction in CPR with an additional vitrification-warming (G1 54.3% vs. G2 47.1%) and larger reduction with an additional embryo biopsy (G2 47.1% vs. G3 31.0%), but neither difference was statistically significant. However, the combined effect of both additional biopsy and vitrification-warming resulted in a significantly reduced CPR (G1 54.3% vs. G3 31.0%; P=.013). This study indicates that blastocysts biopsied and vitrified-warmed twice have reduced clinical outcomes compared with blastocysts biopsied and vitrified-warmed once. PGD patients should be advised that performing a second biopsy and vitrification-warming in cases of failure to obtain a result from initial biopsy will reduce the chance of pregnancy. Patients with inherited disorders may elect to proceed with the second biopsy and vitrification to avoid transfer of embryos with the genetic condition. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  5. Effect of Vitrification on Sperm Parameters and Apoptosis in Fertile Men

    Directory of Open Access Journals (Sweden)

    M Adib

    2011-01-01

    Full Text Available Introduction & Objective: Today, cryopreservation of the human sperm is a common technique for treating infertility. It has been indicated that cryopreservation by different methods decrease the sperm motility and viability in fertile men, but still effect of freezing of the sperm by vitrification method have not been evaluated on sperm parameters and apoptosis. The aim of this study was to evaluate the effect of vitrification of sperm of fertile men on different sperm parameters (motility, morphology, viability and count and apoptosis after thawing. Materials & Methods: In this experimental study which was conducted at Yazd Infertility Research and Clinical Center in 2009, seventeen semen samples were collected by masturbation from people who came to this centre. Semen analysis was performed according to WHO standards. Smear was provided from these samples and fixed for TUNEL staining. Some samples were directly cryopreserved by cryoloope in liquid nitrogen and stored at least for Seven days. After thawing, samples were evaluated for sperm parameters. The collected data was analyzed by the SPSS software using paired T-test and Willcoxon statistical test. Results: The progressive movement of sperm was significantly decreased by vitrification. Also significant decrease in viability and morphology of the sperm and increase in the rate of apoptosis was observed after vitrification. The amount of apoptosis had negatively correlated with normal parameters of spermatozoa (especially progressive motility and viability. Conclusion: These results indicated that vitrification is harmful for sperm parameters and of apoptosis rate in fertile men. However, the apoptosis rate was lower compared to other freezing methods.

  6. Effect of vitrification using the Cryotop method on the gene expression profile of in vitro-produced bovine embryos.

    Science.gov (United States)

    de Oliveira Leme, Ligiane; Dufort, Isabelle; Spricigo, José Felipe Warmling; Braga, Thiago Felipe; Sirard, Marc-André; Franco, Maurício Machaim; Dode, Margot Alves Nunes

    2016-03-01

    The present study analyzed the changes in gene expression induced by the Cryotop vitrification technique in bovine blastocyst-stage embryos, using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos were vitrified and compared with nonvitrified (control) embryos. After vitrification, embryos were warmed and cultured for an additional 4 hours. Survived embryos were used for microarray analysis and quantitative polymerase chain reaction (qPCR) quantification. Survival rates were higher (P vitrification seems to be the activation of the apoptosis pathway in some cells. Indeed, FOSL1 is part of the activating protein 1 transcription factor complex and is implicated in a variety of cellular processes, including proliferation, differentiation, and apoptosis. Therefore, our results suggest that a limited increase in the rate of apoptosis was the only detectable response of the embryos to vitrification stress. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. What is the net effect of introducing vitrification for cryopreservation of surplus 2PN oocytes in an IVF program?

    Science.gov (United States)

    Golakov, Manja; Depenbusch, Marion; Schultze-Mosgau, Askan; Schoepper, Beate; Hajek, Jennifer; Neumann, Kay; Griesinger, Georg

    2018-02-01

    The aim of this study was to accurately describe outcome differences (cryo-survival, pregnancy rate and live birth rate, both per ET and cumulatively), between the vitrification method and slow-freezing method of surplus 2PN oocytes in an IVF program. In 2004, the freezing method for 2PN oocytes was changed from slow-cooling to vitrification. The data of 711 patients (timespan: 1/1999-7/2011; 410 vitrification and 301 slow-cooling events) undergoing a first IVF/ICSI cycles with freezing of 2PN oocytes were retrospectively analyzed. The outcome of one, the first, IVF cycle per patient was explored. The data were analyzed per freezing-thawing attempt as well as cumulatively per one complete IVF cycle, taking pregnancy occurrence after a fresh embryo transfer preceding the cryo-cycle(s) and other confounders (such as female age, elective vs. surplus 2PN cryopreservation) into account by means of exploratory regression analyses. In the vitrification and slow-cooling group, 756 and 376, respectively, attempts of thawing 2PN oocytes were recorded. Each attempt of thawing 2PN oocytes showed statistically significantly higher mean cryo-survival rates after vitrification (effect size approximately 30-40%, with vitrification cryo-survival consistently above 90% in all thawing attempts). Furthermore, the incidence of "zero survival" was lower after vitrification (0.5 vs. 7.3%, p IVF cycle (fresh and frozen transfers combined) with vitrification of 2PN oocytes is increased approximately 1.4-fold (OR of 1.405, 95% CI 0.968-2.038; p = 0.07); however, statistical significance was not achieved due to sample size. Female age and elective cryopreservation of all 2PN oocytes without a fresh transfer (e.g., hyperresponders) were found to be negatively and positively, respectively, associated with the chance of achieving a live birth. The introduction of vitrification has a measurable impact on the efficacy of an IVF program. However, this effect is not large despite the

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

    Science.gov (United States)

    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 (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

  9. APPLICATION OF MECHANICAL ACTIVATION TO PRODUCTION OF PYROCHLORE CERAMIC CONTAINING SIMULATED RARE-EARTH ACTINIDE FRACTION OF HLW

    Energy Technology Data Exchange (ETDEWEB)

    Stefanovsky, S.V.; Kirjanova, O.I.; Chizhevskaya, S.V.; Yudintsev, S.V.; Nikonov, B.S.

    2003-02-27

    Samples of zirconate pyrochlore ceramic (REE)2(Zr,U)2O7 (REE = La-Gd) containing simulated REE-An fraction of HLW were synthesized by two routes: (1) conventional cold compaction of oxide mixtures in pellets under pressure of 200 MPa and sintering of the pellets at 1550 C for 24 hours; and (2) using preliminary mechanical activation of oxide powders in a linear inductive rotator (LIV-0.5E) and a planetary mill - activator with hydrostatic yokes (AGO-2U) for 5 or 10 min. All the samples sintered at 1550 C were monolithic and dense with high mechanical integrity. As follows from X-ray diffraction (XRD) data, the ceramic sample produced without mechanical activation is composed of pyrochlore as major phase but contains also minor unreacted oxides. The samples prepared from pre-activated mixtures are composed of the pyrochlore structure phase only. Scanning electron microscopy (SEM) data also show higher structural and compositional homogeneity of the samples prepared from mechanically activated batches. The samples produced from oxide mixtures mechanically activated in the LIV for 10 min were slightly contaminated with iron resulting in formation of minor perovskite structure phase not detected by XRD but seen on SEM-images of the samples. Comparison of the samples prepared from non-activated and activated batches showed higher density, lower open porosity, water uptake, and elemental leaching for the samples fabricated from mechanically activated oxide mixtures.

  10. Vitrification of in vivo and in vitro produced ovine blastocysts.

    Science.gov (United States)

    Zhu, S E; Zeng, S M; Yu, W L; Li, S J; Zhang, Z C; Chen, Y F

    2001-11-01

    Although cryopreservation of bovine embryo has made great progress in recent years, little achievement was obtained in ovine embryo freezing, especially in vitro produced embryos. However, a simple and efficient method for cryopreservation of sheep embryos will be important for application of ovine embryonic techniques such as in vitro fertilization, transgenic, cloning and etc. In this study ovine blastocysts, produced in vivo or in vitro, were cryopreserved by vitrification in EFS40 (40% ethylene glycol (EG), 18% ficoll and 0.5 M sucrose) or GFS40 (40% glycerol (GL), 18% ficoll and 0.5 Mol sucrose). In vitro produced, early blastocysts were directly plunged into liquid nitrogen (LN2) after preparation by one of the following procedures at 25 degrees C: (A) equilibration in EFS40 for 1 min; (B) equilibration in EFS40 for 2 min; (C) equilibration in EFS40 for 30 s following pretreatment in 10% EG for 5 min; (D) equilibration in EFS40 for 30 s following pretreatment in EFS20 for 2 min (E) equilibration in GFS30 for 30 s following pretreatment in 10% GL for 5 min. The survival rates observed after thawing and in vitro culture for 12 h were A 78.0% (39/50), B 50.0% (26/52), C 93.3% (70/75), D 92.0% (46/50) and E 68.0% (34/50). Survival rates were not significantly different for treatments C and D (p>0.05), but those for groups C and D were significantly higher than for A, B and E (pvitro culture, hatched blastocyst rates were A 28.0% (14/50), B 21.1% (11/52), C 49.3% (37/75), D 48.0% (24/50), E 32.0% (16/50) and control 54.0% (27/50). The hatching rates for groups A, B and E were significantly lower than the control (p0.05). The freezing procedures A, B and C were used to vitrify in vivo produced, early blastocysts recovered from superovulated ewes. The survival rates of frozen-thawed in vivo embryos were A 94.7% (72/76), B 75.0% (45/60) and C 96.4% (54/56) and for group B was significantly lower than for the other two treatment groups (p0.05). Frozen-thawed embryos

  11. Asouzu's Complementary Ontology as a Foundation for a Viable ...

    African Journals Online (AJOL)

    This paper on “Asouzu's Complementary Ontology as a foundation for a viable Ethic of the Environment”, posits that an ethic of the environment can be seen as viable if it considers the whole of reality as ontologically relevant. This point of view would free environmental ethics of anthropocentric bias and its attendant ...

  12. Hanford Waste Vitrification Plant Quality Assurance Program description for high-level waste form development and qualification. Revision 3, Part 2

    Energy Technology Data Exchange (ETDEWEB)

    1993-08-01

    The Hanford Waste Vitrification Plant Project has been established to convert the high-level radioactive waste associated with nuclear defense production at the Hanford Site into a waste form suitable for disposal in a deep geologic repository. The Hanford Waste Vitrification Plant will mix processed radioactive waste with borosilicate material, then heat the mixture to its melting point (vitrification) to forin a glass-like substance that traps the radionuclides in the glass matrix upon cooling. The Hanford Waste Vitrification Plant Quality Assurance Program has been established to support the mission of the Hanford Waste Vitrification Plant. This Quality Assurance Program Description has been written to document the Hanford Waste Vitrification Plant Quality Assurance Program.

  13. Survival and development of bovine blastocysts produced in vitro after assisted hatching, vitrification and in-straw direct rehydration.

    Science.gov (United States)

    Vajta, G; Holm, P; Greve, T; Callesen, H

    1997-09-01

    The purpose of this study was to establish an efficient combination of assisted hatching and cryopreservation procedures for producing bovine embryos in vitro. A total of 1312 day 7 blastocysts were subjected randomly to 14 different combinations of three factors: osmotic stress, assisted hatching and vitrification. Re-expansion, initiation and completion of the hatching process, as well as attachment to the culture dish, were analysed by SAS Genmod procedure. Incubation with sucrose was found to decrease survival rates; among the assisted hatching procedures used, zona fenestration resulted in higher survival rates compared with partial zona dissection and controls; and vitrification decreased survival and further development. The combined effect of sucrose incubation and vitrification decreased further development markedly, as did partial zona dissection followed by vitrification. Partial zona dissection performed in medium containing sucrose severely lowered embryo survival. Zona fenestration without sucrose incubation followed by vitrification did not compromise further embryo development: 86%, 84% and 79% of the blastocysts initiated, completed hatching and attached to the bottom, respectively. These data were not different from the controls (80%, 76% and 63%, respectively; P > 0.05). Cell count analysis revealed a decrease in the total number of cells as a result of the assisted hatching and vitrification compared with controls (135 versus 202, respectively; P embryo transfer results (36% pregnancy rate and 30% calving rate) require further improvement, this combination of methods may prove useful in the commercial production of bovine embryos in vitro.

  14. Vitrification of immature bovine cumulus-oocyte complexes: effects of cryoprotectants, the vitrification procedure and warming time on cleavage and embryo development.

    Science.gov (United States)

    Prentice-Biensch, Jennifer R; Singh, Jaswant; Mapletoft, Reuben J; Anzar, Muhammad

    2012-09-06

    The present studies evaluated the effects of cryoprotectants, the vitrification procedure and time in the warming solution containing sucrose on cleavage and embryo development of immature (GV stage) bovine cumulus-oocyte complexes (COCs). Two experiments were conducted. In Experiment 1, COCs (n = 420) were randomly assigned to four groups: 1) CONTROL GROUP: no treatment; 2) VS1 group: COCs were exposed to vitrification solution 1 (VS1) containing 7.5% ethylene glycol [EG] + 7.5% dimethyl sulfoxide [DMSO] + 20% calf serum [CS] in TCM-199 at 37 C for 5 min; 3) VS1 + VS2 group: COCs were exposed to VS1 for 5 min followed by VS2 (15% EG + 15% DMSO + 17.1% sucrose + 20% CS) at 37 C for 45-60 sec; and 4) Vitrified group: COCs were exposed to VS1 and VS2, loaded on cryotops, vitrified in liquid nitrogen and then warmed in TCM-199 + 17.1% sucrose + 20% CS at 37 C for 1 min. In Experiment 2, COCs (n = 581) were assigned to the same groups, but those in VS1, VS1 + VS2 and Vitrified groups were sub-divided and exposed to the warming solution for either 1 or 5 min. After treatment and/or warming, all COCs in both experiments underwent in vitro maturation, in vitro fertilization and in vitro culture. Cleavage and blastocyst rates did not differ among Control, VS1 and VS1 + VS2 groups in either experiment. In Experiment 2, there was no effect of time in the warming solution.However, both cleavage and blastocyst rates were lower (P bovine COCs. However, cleavage rate and early embryo development were reduced following the vitrification and warming.

  15. Separation of viable and non-viable tomato (Solanum lycopersicum L.) seeds using single seed near-infrared spectroscopy

    DEFF Research Database (Denmark)

    Shrestha, Santosh; Deleuran, Lise Christina; Gislum, René

    2017-01-01

    -viable tomato seeds of two cultivars using chemometrics. The data exploration were performed by principal component analysis (PCA). Subsequently, viable and non-viable seeds were classified by partial least squares-discriminant analysis (PLS-DA) and interval PLS-DA (iPLS-DA). The indication of clustering...... of viable and non-viable seeds were observed in the PCA of each cultivar and the pooled samples. However, the PCA did not exhibit a pattern of separation among the early, normal and late germinated tomato seeds. The NIR spectral regions of 1160–1170, 1383–1397, 1647–1666, 1860–1884 and 1915–1940 nm were...... identified as important for classification of viable and non-viable tomato seeds by iPLS-DA. The sensitivity i.e. ability to correctly identify the positive samples and specificity i.e. ability to reject the negative samples of the (iPLS-DA) model on identified spectral regions for prediction of viable...

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

    National Research Council Canada - National Science Library

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

    ... actions.The minimal concentration of ethylene glycol (EG) on mouse oocyte survival and subsequent embryonic development was evaluated following vitrification-warming and parthenogenetic activation...

  17. The nature of the volatile technetium species formed during vitrification of borosilicate glass

    Energy Technology Data Exchange (ETDEWEB)

    Childs, Bradley C.; Poineau, Frederic; Czerwinski, Kenneth R.; Sattelberger, Alfred P.

    2015-05-26

    Vitrification of sodium pertechnetate into borosilicate glass was performed in air at 1100 C. A glass with a composition similar to the one developed for vitrification of the low activity waste at the Hanford site was used. A red volatile species was observed above 600° C. The extended X-ray absorption fine structure results indicate the environment of the absorbing Tc atom consists of 2.9(6) O atoms at 1.73(2) A° , 2.2(4) O atoms at 2.02(2) A° , and 0.8(2) O atoms at 2.18(2) A° . The results are consistent with the presence of a mononuclear species with a structure closely related to TcO3(OH)(H2O)2.

  18. Development of the vitrification compositional envelope to support complex-wide application of MAWS technology

    Energy Technology Data Exchange (ETDEWEB)

    Mazer, J.J. [ed.] [Argonne National Lab., IL (United States); Muller, I.S.; Gan, H.; Buechele, A.C.; Lai, S.T.; Pegg, I.L. [Catholic Univ. of America, Washington, DC (United States). Vitreous State Lab.]|[GTS Duratek, Inc., Columbia, MD (United States)

    1996-09-01

    This report presents the results from a study of the application of the Minimum Additive Waste Stabilization (MAWS) approach using vitrification as a treatment technology to a variety of waste streams across the DOE complex. This work has involved both experimental vitrification work using actual mixed wastes and surrogate waste streams from several DOE sites (Hanford, Idaho, and Oak Ridge) as well as the development of a computer-based, integrated glass property-composition database. The long-term objective is that this data base will assist glass formulation studies with single waste streams or combinations of waste streams subject to a variety of user-imposed constraints including waste stream usage priorities, process related constraints (e.g., melt viscosity, electrical conductivity, etc.), and waste form performance related constraints (e.g., TCLP and PCT leaching results). 79 refs., 143 figs., 65 tabs.

  19. Hanford tank waste simulants specification and their applicability for the retrieval, pretreatment, and vitrification processes

    Energy Technology Data Exchange (ETDEWEB)

    GR Golcar; NG Colton; JG Darab; HD Smith

    2000-04-04

    A wide variety of waste simulants were developed over the past few years to test various retrieval, pretreatment and waste immobilization technologies and unit operations. Experiments can be performed cost-effectively using non-radioactive waste simulants in open laboratories. This document reviews the composition of many previously used waste simulants for remediation of tank wastes at the Hanford reservation. In this review, the simulants used in testing for the retrieval, pretreatment, and vitrification processes are compiled, and the representative chemical and physical characteristics of each simulant are specified. The retrieval and transport simulants may be useful for testing in-plant fluidic devices and in some cases for filtration technologies. The pretreatment simulants will be useful for filtration, Sr/TRU removal, and ion exchange testing. The vitrification simulants will be useful for testing melter, melter feed preparation technologies, and for waste form evaluations.

  20. Bovine oocyte vitrification before or after meiotic arrest: effects on ultrastructure and developmental ability

    OpenAIRE

    Díez, Carmen; Duque, Paloma; Gómez, Enrique; Hidalgo, C.O. (Carlos); Tamargo, Carolina; Rodríguez, Aida; Fernández, Lina; Varga, Santiago; Fernández, Alba; Facal, Nieves; Carbajo, Maite

    2011-01-01

    The nuclear stage at which oocytes are cryopreserved influences further development ability and cryopreservation affects ultrastructure of both cumulus cells and the oocyte. In this work, we analyze the effects of vitrification at different nuclear and cytoplasmic maturation stages on the oocyte ultrastructure and developmental ability. Culture in TCM199 + PVA with roscovitine 25 M during 24 h led to meiotic arrest (MA) in cumulus-oocyte complexes (COCs), while permissive in vitro...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  3. Technology evaluation report: Babcock and Wilcox Cyclone Furnace Vitrification technology. Volume 2

    Energy Technology Data Exchange (ETDEWEB)

    Groeber, P.

    1992-09-01

    The Babcock and Wilcox (B and W) Cyclone Furnace Vitrification Technology is a treatment process for contaminated soils. The process was evaluated to determine its ability to destroy semivolatile organics and to isolate metals and simulated radionuclides into a non-leachable slag material. The feed material for the system was a prepared synthetic soil matrix (SSM) that was spiked with two organic compounds and six metals. This volume contains the appendices.

  4. Hanford Waste Vitrification Plant technical background document for toxics best available control technology demonstration

    Energy Technology Data Exchange (ETDEWEB)

    None

    1992-10-01

    This document provides information on toxic air pollutant emissions to support the Notice of Construction for the proposed Hanford Waste Vitrification Plant (HWVP) to be built at the the Department of Energy Hanford Site near Richland, Washington. Because approval must be received prior to initiating construction of the facility, state and federal Clean Air Act Notices of construction are being prepared along with necessary support documentation.

  5. Removal of Aerosol Particles Generated from Vitrification Process for High-Level Liquid Wastes

    OpenAIRE

    加藤 功

    1990-01-01

    The vitrification technology has been developed for the high-level liquid waste (HLLW) from reprocessing nuclear spent fuel in PNC. The removal performance of the aerosol particles generated from the melting process was studied in a nonradioactive full-scale mock-up test facility (MTF). The off-gas treatment system consists of submerged bed scrubber (SBS), venturi scrubber, NOx absorber, high efficiency mist eliminater (HEME). Deoomtamination factors (DFs) were derived from the mass ratio of ...

  6. Development of Vitrification Process and Glass Formulation for Nuclear Waste Conditioning

    Energy Technology Data Exchange (ETDEWEB)

    Petitjean, V.; Fillet, C.; Boen, R.; Veyer, C.; Flament, T.

    2002-02-26

    The vitrification of high-level waste is the internationally recognized standard to minimize the impact to the environment resulting from waste disposal as well as to minimize the volume of conditioned waste to be disposed of. COGEMA has been vitrifying high-level waste industrially for over 20 years and is currently operating three commercial vitrification facilities based on a hot metal crucible technology, with outstanding records of safety, reliability and product quality. To further increase the performance of vitrification facilities, CEA and COGEMA have been developing the cold crucible melter technology since the beginning of the 1980s. This type of melter is characterized by a virtually unlimited equipment service life and a great flexibility in dealing with various types of waste and allowing development of high temperature matrices. In complement of and in parallel with the vitrification process, a glass formulation methodology has been developed by the CEA in order to tailor matrices for the wastes to be conditioned while providing the best adaptation to the processing technology. The development of a glass formulation is a trade-off between material properties and qualities, technical feasibility, and disposal safety criteria. It involves non-radioactive and radioactive laboratories in order to achieve a comprehensive matrix qualification. Several glasses and glass ceramics have thus been studied by the CEA to be compliant with industrial needs and waste characteristics: glasses or other matrices for a large spectrum of fission products, or for high contents of specifics elements such as sodium, phosphate, iron, molybdenum, or actinides. New glasses or glass-ceramics designed to minimize the final wasteform volume for solutions produced during the reprocessing of high burnup fuels or to treat legacy wastes are now under development and take benefit from the latest CEA hot-laboratories and technology development. The paper presents the CEA state

  7. Selection of melter systems for the DOE/Industrial Center for Waste Vitrification Research

    Energy Technology Data Exchange (ETDEWEB)

    Bickford, D.F.

    1993-12-31

    The EPA has designated vitrification as the best developed available technology for immobilization of High-Level Nuclear Waste. In a recent federal facilities compliance agreement between the EPA, the State of Washington, and the DOE, the DOE agreed to vitrify all of the Low Level Radioactive Waste resulting from processing of High Level Radioactive Waste stored at the Hanford Site. This is expected to result in the requirement of 100 ton per day Low Level Radioactive Waste melters. Thus, there is increased need for the rapid adaptation of commercial melter equipment to DOE`s needs. DOE has needed a facility where commercial pilot scale equipment could be operated on surrogate (non-radioactive) simulations of typical DOE waste streams. The DOE/Industry Center for Vitrification Research (Center) was established in 1992 at the Clemson University Department of Environmental Systems Engineering, Clemson, SC, to address that need. This report discusses some of the characteristics of the melter types selected for installation of the Center. An overall objective of the Center has been to provide the broadest possible treatment capability with the minimum number of melter units. Thus, units have been sought which have broad potential application, and which had construction characteristics which would allow their adaptation to various waste compositions, and various operating conditions, including extreme variations in throughput, and widely differing radiological control requirements. The report discusses waste types suitable for vitrification; technical requirements for the application of vitrification to low level mixed wastes; available melters and systems; and selection of melter systems. An annotated bibliography is included.

  8. Good Preservation of Stromal Cells and No Apoptosis in Human Ovarian Tissue after Vitrification

    OpenAIRE

    Raffaella Fabbri; Rossella Vicenti; Maria Macciocca; Gianandrea Pasquinelli; Roberto Paradisi; Cesare Battaglia; Nicola Antonio Martino; Stefano Venturoli

    2014-01-01

    The aim of this study was to develop a vitrification procedure for human ovarian tissue cryopreservation in order to better preserve the ovarian tissue. Large size samples of ovarian tissue retrieved from 15 female-to-male transgender subjects (18–38 years) were vitrified using two solutions (containing propylene glycol, ethylene glycol, and sucrose at different concentrations) in an open system. Light microscopy, transmission electron microscopy, and TUNEL assay were applied to evaluate the ...

  9. Vitrification of bovine embryos followed by in vitro hatching and expansion.

    Science.gov (United States)

    Souza, J F; Oliveira, C M; Lienou, L L; Cavalcante, T V; Alexandrino, E; Santos, R R; Rodrigues, A P R; Campello, C C; Figueiredo, J R; Dias, F E F

    2017-12-18

    The objective of this study was to assess the effects of bovine embryo vitrification by applying three different vitrification solutions containing ethylene glycol (EG) and dimethylsulphoxide (DMSO) at different concentrations (10, 20 or 25% each) combined with 1.0 M glucose or 1.0 M sucrose, on the in vitro hatching and expansion rates. Healthy oocytes were selected for in vitro maturation and fertilization from 200 bovine ovaries, and subsequently cultured up to the blastocyst stage (n = 800). Control (n = 200) and vitrified cells (n = 100 per treatment; 600 in total) were cultured for an extra 24 or 48 h to evaluate hatching and expansion, respectively. Vitrification significantly decreased embryonic re-expansion and hatching rates independently of the tested solution when compared with control embryos, but solutions with 25% EG + 25% DMSO resulted in the highest re-expansion (75%) and hatching (70%) rates, independently of the added sugar. The addition of sucrose resulted in higher rates of re-expanded and hatched embryos when compared with glucose addition. We concluded that the combination of 25% EG + 25% DMSO and 1.0 M sucrose allowed hatching and expansion of vitrified-warmed bovine embryos produced in vitro.

  10. The beneficial effects of antifreeze proteins in the vitrification of immature mouse oocytes.

    Directory of Open Access Journals (Sweden)

    Jun Woo Jo

    Full Text Available Antifreeze proteins (AFPs are a class of polypeptides that permit organismal survival in sub-freezing environments. The purpose of this study was to investigate the effect of AFP supplementation on immature mouse oocyte vitrification. Germinal vesicle-stage oocytes were vitrified using a two-step exposure to equilibrium and vitrification solution in the presence or absence of 500 ng/mL of AFP III. After warming, oocyte survival, in vitro maturation, fertilization, and embryonic development up to the blastocyst stage were assessed. Spindle and chromosome morphology, membrane integrity, and the expression levels of several genes were assessed in in vitro matured oocytes. The rate of blastocyst formation was significantly higher and the number of caspase-positive blastomeres was significantly lower in the AFP-treated group compared with the untreated group. The proportion of oocytes with intact spindles/chromosomes and stable membranes was also significantly higher in the AFP group. The AFP group showed increased Mad2, Hook-1, Zar1, Zp1, and Bcl2 expression and lower Eg5, Zp2, Caspase6, and Rbm3 expression compared with the untreated group. Supplementation of the vitrification medium with AFP has a protective effect on immature mouse oocytes, promoting their resistance to chilling injury. AFPs may preserve spindle forming ability and membrane integrity at GV stage. The fertilization and subsequent developmental competence of oocytes may be associated with the modulation of Zar1, Zp1/Zp2, Bcl2, Caspase6, and Rbm3.

  11. Test Summary Report INEEL Sodium-Bearing Waste Vitrification Demonstration RSM-01-1

    Energy Technology Data Exchange (ETDEWEB)

    Goles, Ronald W.; Perez, Joseph M.; Macisaac, Brett D.; Siemer, Darryl D.; Mccray, John A.

    2001-05-21

    The U.S. Department of Energy's Idaho National Engineering and Environmental Laboratory is storing large amounts of radioactive and mixed wastes. Most of the sodium-bearing wastes have been calcined, but about a million gallons remain uncalcined, and this waste does not meet current regulatory requirements for long-term storage and/or disposal. As a part of the Settlement Agreement between DOE and the State of Idaho, the tanks currently containing SBW are to be taken out of service by December 31, 2012, which requires removing and treatment the remaining SBW. Vitrification is the option for waste disposal that received the highest weighted score against the criteria used. Beginning in FY 2000, the INEEL high-level waste program embarked on a program for technology demonstration and development that would lead to conceptual design of a vitrification facility in the event that vitrification is the preferred alternative for SBW disposal. The Pacific Northwest National Laborator's Research-Scale Melter was used to conduct these initial melter-flowsheet evaluations. Efforts are underway to reduce the volume of waste vitrified, and during the current test, an overall SBW waste volume-reduction factor of 7.6 was achieved.

  12. Optimization of a novel nylon mesh container for human embryo ultrarapid vitrification.

    Science.gov (United States)

    Nakashima, Akira; Ino, Nao; Kusumi, Maki; Ohgi, Shirei; Ito, Megumu; Horikawa, Takashi; Nakagawa, Koji; Saito, Takakazu; Kamura, Toshiharu; Saito, Hidekazu

    2010-05-01

    To evaluate the efficacy of a nylon mesh container in vitrification of human embryos and to determine the optimal osmotic pressure of the initial thawing solution. Retrospective analysis. National Center for Child Health and Development, Tokyo, Japan. Infertile patients undergoing either in vitro fertilization or intracytoplasmic sperm injection in our hospital. Embryos, at the cleavage stage, were cryopreserved using the vitrification method in either a plastic straw or a nylon mesh container. The embryos were thawed using an initial osmotic pressure of either 0.5 M or 1.0 M sucrose with subsequent step-wise dilution. After thawing, the embryos were transferred to the uterus. Survival rate of blastomeres, embryo survival rate, implantation, and pregnancy rates, cancellation rate because of embryo damage. Use of nylon mesh and the 1.0 M sucrose thawing solution significantly improved blastomere survival rate (98.0 +/- 1.0%, mean +/- SEM), pregnancy rate (41.0%) and implantation rate (32.3%). Vitrification using a nylon mesh container and subsequent thawing in a 1.0 M sucrose solution is an easy and inexpensive method that improves the reliability of embryo cryopreservation of embryos without adverse effects on clinical outcomes. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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.

  14. Effects of various freezing containers for vitrification freezing on mouse oogenesis.

    Science.gov (United States)

    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 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. Demonstration plasma gasification/vitrification system for effective hazardous waste treatment.

    Science.gov (United States)

    Moustakas, K; Fatta, D; Malamis, S; Haralambous, K; Loizidou, M

    2005-08-31

    Plasma gasification/vitrification is a technologically advanced and environmentally friendly method of disposing of waste, converting it to commercially usable by-products. This process is a drastic non-incineration thermal process, which uses extremely high temperatures in an oxygen-starved environment to completely decompose input waste material into very simple molecules. The intense and versatile heat generation capabilities of plasma technology enable a plasma gasification/vitrification facility to treat a large number of waste streams in a safe and reliable manner. The by-products of the process are a combustible gas and an inert slag. Plasma gasification consistently exhibits much lower environmental levels for both air emissions and slag leachate toxicity than other thermal technologies. In the framework of a LIFE-Environment project, financed by Directorate General Environment and Viotia Prefecture in Greece, a pilot plasma gasification/vitrification system was designed, constructed and installed in Viotia Region in order to examine the efficiency of this innovative technology in treating industrial hazardous waste. The pilot plant, which was designed to treat up to 50kg waste/h, has two main sections: (i) the furnace and its related equipment and (ii) the off-gas treatment system, including the secondary combustion chamber, quench and scrubber.

  16. Vitrification of ICSI- and IVF-derived bovine blastocysts by minimum volume cooling procedure: effect of developmental stage and age.

    Science.gov (United States)

    Abdalla, H; Shimoda, M; Hara, H; Morita, H; Kuwayama, M; Hirabayashi, M; Hochi, S

    2010-10-01

    The objective was to investigate the effects of developmental stage (fully-expanded or expanding blastocysts) and/or age (harvested on Days 7 or 8) on post-vitrification in vitro survival of bovine blastocysts derived from intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Post-warming survival (re-expansion of blastocoele within 24 h) of ICSI-derived fully-expanded blastocysts (80%) was similar to that of their IVF-derived counterparts (88%). However, the ability of ICSI-derived expanding blastocysts to survive vitrification procedures (61%) was lower than that of IVF-derived blastocysts (85%; P vitrification did not affect cryotolerance for either ICSI-derived (73 and 59% for Days 7 and 8 embryos, respectively) or IVF-derived blastocysts (86% for both Days 7 and 8 embryos). At 24 h of post-warming culture, ICSI-derived blastocysts surviving vitrification contained a higher proportion of dead cells than their IVF-derived counterparts (5-13% vs. 2-4%; P vitrification on the ability of blastocysts to hatch within 72 h of culture only in IVF-derived Day 8 blastocysts (41 and 70% in vitrified and fresh control groups, respectively). In conclusion, the proportion of blastocysts that survived vitrification procedures was similar for ICSI- and IVF-derived bovine blastocysts if the former were cultured to the fully-expanded stage prior to vitrification, with no significant difference between embryos harvested on Day 7 versus Day 8. (c) 2010 Elsevier Inc. All rights reserved.

  17. Viable Cell Culture Banking for Biodiversity Characterization and Conservation.

    Science.gov (United States)

    Ryder, Oliver A; Onuma, Manabu

    2018-02-15

    Because living cells can be saved for indefinite periods, unprecedented opportunities for characterizing, cataloging, and conserving biological diversity have emerged as advanced cellular and genetic technologies portend new options for preventing species extinction. Crucial to realizing the potential impacts of stem cells and assisted reproductive technologies on biodiversity conservation is the cryobanking of viable cell cultures from diverse species, especially those identified as vulnerable to extinction in the near future. The advent of in vitro cell culture and cryobanking is reviewed here in the context of biodiversity collections of viable cell cultures that represent the progress and limitations of current efforts. The prospects for incorporating collections of frozen viable cell cultures into efforts to characterize the genetic changes that have produced the diversity of species on Earth and contribute to new initiatives in conservation argue strongly for a global network of facilities for establishing and cryobanking collections of viable cells.

  18. Worst-Case" Simulant for INTEC Soduim-Bearing Waste Vitrification Tests

    Energy Technology Data Exchange (ETDEWEB)

    Christian, Jerry Dale; Batcheller, Thomas Aquinas

    2001-09-01

    The Idaho Nuclear Technology and Engineering Center (INTEC) is developing technologies to process the radioactive liquid sodium-bearing waste from the waste tanks at INTEC to solidify the waste into a form suitable for disposition in a National high-level waste repository currently being considered at Yucca Mountain, Nevada. The requirement is for a qualified glass waste form. Therefore, vitrification is being developed using laboratory, research-scale, and pilot scale melters. While some laboratory experiments can be done with actual waste, the larger scale and most laboratory experiments must be done on non-radioactive simulant waste solutions. Some tests have previously been done on simulants of a representative waste that has been concentrated and will remain unchanged in tank WM-180 until it is vitrified. However, there is a need to develop glass compositions that will accommodate all future wastes in the tanks. Estimates of those future waste compositions have been used along with current compositions to develop a “worst-case” waste composition and a simulant preparation recipe suitable for developing a bracketing glass formulation and for characterizing the flowpath and decontamination factors of pertinent off-gas constituents in the vitrification process. The considerations include development of criteria for a worst-case composition. In developing the criteria, the species that are known to affect vitrification and glass properties were considered. Specific components that may need to be characterized in the off-gas cleanup system were considered in relation to detection limits that would need to be exceeded in order to track those components. Chemical aspects of various constituent interactions that should be taken into account when a component may need to be increased in concentration from that in the actual waste for detection in experiments were evaluated. The worst-case waste simulant composition is comprised of the highest concentration of each

  19. DATA SUMMARY REPORT SMALL SCALE MELTER TESTING OF HLW ALGORITHM GLASSES MATRIX1 TESTS VSL-07S1220-1 REV 0 7/25/07

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; PEGG IL

    2011-12-29

    Eight tests using different HLW feeds were conducted on the DM100-BL to determine the effect of variations in glass properties and feed composition on processing rates and melter conditions (off-gas characteristics, glass processing, foaming, cold cap, etc.) at constant bubbling rate. In over seven hundred hours of testing, the property extremes of glass viscosity, electrical conductivity, and T{sub 1%}, as well as minimum and maximum concentrations of several major and minor glass components were evaluated using glass compositions that have been tested previously at the crucible scale. Other parameters evaluated with respect to glass processing properties were +/-15% batching errors in the addition of glass forming chemicals (GFCs) to the feed, and variation in the sources of boron and sodium used in the GFCs. Tests evaluating batching errors and GFC source employed variations on the HLW98-86 formulation (a glass composition formulated for HLW C-106/AY-102 waste and processed in several previous melter tests) in order to best isolate the effect of each test variable. These tests are outlined in a Test Plan that was prepared in response to the Test Specification for this work. The present report provides summary level data for all of the tests in the first test matrix (Matrix 1) in the Test Plan. Summary results from the remaining tests, investigating minimum and maximum concentrations of major and minor glass components employing variations on the HLW98-86 formulation and glasses generated by the HLW glass formulation algorithm, will be reported separately after those tests are completed. The test data summarized herein include glass production rates, the type and amount of feed used, a variety of measured melter parameters including temperatures and electrode power, feed sample analysis, measured glass properties, and gaseous emissions rates. More detailed information and analysis from the melter tests with complete emission chemistry, glass durability, and

  20. Vitrification of cleavage stage day 3 embryos results in higher live birth rates than conventional slow freezing: a RCT.

    Science.gov (United States)

    Debrock, S; Peeraer, K; Fernandez Gallardo, E; De Neubourg, D; Spiessens, C; D'Hooghe, T M

    2015-08-01

    Is the live birth rate (LBR) per embryo thawed/warmed higher when Day 3 cleavage stage embryos are cryopreserved by vitrification compared with slow freezing? The LBR per embryo thawed/warmed was higher after vitrification than after slow freezing on Day 3, based on better embryo survival, quality and availability of embryos in the vitrification group. Post-thawing survival rate of cleavage-stage embryos has been reported to be higher after vitrification than after slow freezing. This RCT was performed in an academic tertiary center between September 2011 and March 2013. If supernumerary embryos were available on Day 3, patients were randomized at the time of cryopreservation using a computerized system to determine a simple allocation to the vitrification group or the slow freezing group and all embryos were frozen with the same technique. The primary outcome of this study was the LBR per embryo thawed/warmed. Power calculation revealed that 184 thawed embryos were needed in each group (β = 0.8, α freezing (6%). Patients freezing (using 1,2-propanediol and 0.1 M sucrose as cryoprotectant) or by closed vitrification using commercially available freezing/vitrification media. Survival was defined as ≥50% cells were intact after thawing. Thawed embryos were further cultured overnight. In total, 307 patients were randomized to slow freezing (155 patients, 480 embryos) or vitrification (152 patients, 495 embryos). By March 2013, 200 embryos were thawed after slow freezing in 95 cycles for 79 patients and 217 embryos were warmed after vitrification in 121 cycles in 90 patients. The LBR per embryo thawed/warmed was significantly higher after vitrification (16.1% (35/217)) than after slow freezing (5.0% (10/200); P freezing (7.5% (15/200); P = 0.0012; RR 2.76; CI 1.59-4.81). The survival rate was significantly higher after vitrification (84.3% (183/217) than after slow freezing (52.5% (105/200); P freezing (28.6% (30/105); P freezing (73.7% (70/95); P = 0

  1. Vitrification of bovine oocytes at different meiotic stages using the Cryotop method: assessment of morphological, molecular and functional patterns.

    Science.gov (United States)

    Sprícigo, J F W; Morais, K; Ferreira, A R; Machado, G M; Gomes, A C M; Rumpf, R; Franco, M M; Dode, M A N

    2014-10-01

    This study aimed to investigate the functional, morphological and molecular patterns of bovine oocytes vitrified at different times during in vitro maturation (IVM). Four groups of oocytes were used: non-vitrified control oocytes (CG), oocytes vitrified at 0 h (V0), oocytes vitrified after 8 h of IVM (V8) and oocytes vitrified after 22 h of IVM (V22). After vitrification, the oocytes were warmed and then returned to the incubator to complete a total of 24h of IVM. To evaluate the effect of vitrification, the nuclear maturation and fertilization rates were assessed by lacmoid staining and ultrastructural electron microscopy. The cleavage and blastocyst rates were evaluated at D2, D7 and D8. The expression levels of CASP3, TP53, HDAC2, SUV39H1 and DNMT1 were investigated by RT-qPCR. The nuclear maturation, oocyte fertilization, cleavage and blastocyst rates were higher (P vitrification (P > 0.05). In conclusion, cytoplasm degeneration seems to be the most severe form of damage caused by vitrification. The use of the Cryotop method for vitrification severely reduces bovine oocyte viability regardless of whether it is performed at GV, GVBD or MII stage. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. A SIMPLE AND EFFICIENT VITRIFICATION METHOD FOR IN-STRAW DILUTION AND DIRECT TRANSFER OF BOVINE EMBRYOS.

    Science.gov (United States)

    Zhang, Youwen; Fu, Xiangwei; Chen, Long; Feng, Chuntao; Bi, Jianghua; Mo, Xianhong; Cheng, Keren; Zhang, Rina; Li, Shujing; Zhu, Shien

    2015-01-01

    An easy and user friendly protocol that produces consistent results will facilitate the commercial application of embryo vitrification technology in the field. This study was designed to develop a simple and efficient vitrification, in-straw dilution and direct transfer method for bovine embryos. After being vitrified and in-straw thawed, in vivo-derived and in vitro-produced bovine embryos were subjected to in vitro culture or embryo transplantation. There were no significant differences (P > 0.05) in survival rates (100.0% vs. 93.9%) and expansion rates (93.8% vs. 87.5%) between in vivo-derived and in vitro-produced blastocysts after vitrification and in-straw dilution. And there was also no significant difference (P > 0.05) in conception rates (56.5% vs. 58.8%) after ET between cryopreserved and fresh in vivo-derived blastocysts. Vitrification using EG-based vitrification solution and in-straw dilution with PBS-based diluent is a simple and efficient method for cryopreservation and direct transfer of bovine embryos.

  3. Expression and distribution of cell adhesion-related proteins in bovine parthenogenetic embryos: The effects of oocyte vitrification.

    Science.gov (United States)

    Zeng, Yan; Fu, Xiangwei; Zhou, Guangbin; Yue, Mingxing; Zhou, Yanhua; Zhu, Shien

    2013-07-01

    The objective was to investigate expression of cell adhesion-related proteins (E-cadherin, β-catenin, and the cytoskeletal protein F-actin) in bovine parthenogenetic embryos derived from vitrified-warmed oocytes. Bovine oocytes at metaphase II were randomly allocated into three groups: (1) untreated (control); (2) exposed to vitrification solution without freezing (toxicity); and (3) vitrified and warmed by the open-pulled straw method (vitrification). After parthenogenetic activation, in the vitrification group compared with the control, the timing of compaction was delayed in (108-120 vs. 96-108 hours, respectively), and the percentage of blastocysts that developed from eight-cell embryos was lower (32.08% vs. 61.03%; P vitrification delayed embryo compaction by affecting adhesion junction formation and function, immunostaining and quantitative reverse transcription polymerase chain reaction were done to characterize distribution patterns (E-cadherin, β-catenin, and the cytoskeletal protein F-actin) and expression levels of cell adhesion-related proteins (β-catenin). Distribution of β-catenin in eight-cell embryos from the vitrification group changed dramatically compared with the control and toxicity groups. Relative expression of β-catenin at the mRNA and protein levels was lower (P bovine parthenogenetic eight-cell embryos derived from vitrified-warmed oocytes were associated with embryo compaction and reduced competence for subsequent embryo development. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Three dimensional in vitro culture of preantral follicles following slow-freezing and vitrification of mouse ovarian tissue.

    Science.gov (United States)

    Asgari, Fatemeh; Valojerdi, Mojtaba Rezazadeh; Ebrahimi, Bita; Fatehi, Roya

    2015-12-01

    To evaluate the effects slow-freezing and vitrification on three dimensional in vitro culture of preantral follicles, ovaries of 12-14 days old female NMRI mice were isolated and randomly assigned to fresh control, slow-freezing and vitrification groups. Slow-freezing was performed using programmable freezer. Vitrification was carried out in a medium consisting of ethylene glycol (EG) and dimethyl sulphoxide (Me2SO) by needle immersion method. middle sized preantral follicles were mechanically isolated and cultured for 12 days in 0.7% sodium alginate gel. The follicles development and quantitative expression of oocyte specific genes (Bmp15, Gdf9, Fgf8) and the growth related genes (Igf1, Kit, Kit-l) were assessed after 1, 8 and 12 days of culture. Both cryopreserved groups showed reduction of follicular survival rates compared to the control group on days 8 and 12 of culture (P culture (P culture (P culture (p culture than vitrification and control groups (P culture. Thus, cryopreservation of mouse ovaries by both methods can preserve most developmental parameters and expression of maturation genes. However, vitrification is a better method for cryopreservation of mouse ovaries due to greater antrum formation and expression of growth related markers. Copyright © 2015. Published by Elsevier Inc.

  5. Cryopreservation of human oocytes, zygotes, embryos and blastocysts: A comparison study between slow freezing and ultra rapid (vitrification methods

    Directory of Open Access Journals (Sweden)

    Tahani Al-Azawi

    2013-12-01

    Full Text Available Preservation of female genetics is currently done primarily by means of oocyte and embryo cryopreservation. The field has seen much progress during its four-decade history, progress driven predominantly by research in humans. It can also be done by preservation of ovarian tissue or entire ovary for transplantation, followed by oocyte harvesting or natural fertilization. Two basic cryopreservation techniques rule the field, slow-rate freezing, the first to be developed and vitrification which in recent years, has gained a foothold. The slow-rate freezing method previously reported had low survival and pregnancy rates, along with the high cost of cryopreservation. Although there are some recent data indicating better survival rates, cryopreservation by the slow freezing method has started to discontinue. Vitrification of human embryos, especially at early stages, became a more popular alternative to the slow rate freezing method due to reported comparable clinical and laboratory outcomes. In addition, vitrification is relatively simple, requires no expensive programmable freezing equipment, and uses a small amount of liquid nitrogen for freezing. Moreover, oocyte cryopreservation using vitrification has been proposed as a solution to maintain women’s fertility by serving and freezing their oocytes at the optimal time. The aim of this research is to compare slow freezing and vitrification in cryopreservation of oocytes, zygotes, embryos and blastocysts during the last twelve years. Therefore, due to a lot of controversies in this regard, we tried to achieve an exact idea about the subject and the best technique used.

  6. Vitrification of immature bovine cumulus-oocyte complexes: effects of cryoprotectants, the vitrification procedure and warming time on cleavage and embryo development

    Directory of Open Access Journals (Sweden)

    Prentice-Biensch Jennifer R

    2012-09-01

    Full Text Available Abstract Background The present studies evaluated the effects of cryoprotectants, the vitrification procedure and time in the warming solution containing sucrose on cleavage and embryo development of immature (GV stage bovine cumulus-oocyte complexes (COCs. Methods Two experiments were conducted. In Experiment 1, COCs (n = 420 were randomly assigned to four groups: 1 Control group: no treatment; 2 VS1 group: COCs were exposed to vitrification solution 1 (VS1 containing 7.5% ethylene glycol [EG] + 7.5% dimethyl sulfoxide [DMSO] + 20% calf serum [CS] in TCM-199 at 37 C for 5 min; 3 VS1 + VS2 group: COCs were exposed to VS1 for 5 min followed by VS2 (15% EG + 15% DMSO + 17.1% sucrose + 20% CS at 37 C for 45–60 sec; and 4 Vitrified group: COCs were exposed to VS1 and VS2, loaded on cryotops, vitrified in liquid nitrogen and then warmed in TCM-199 + 17.1% sucrose + 20% CS at 37 C for 1 min. In Experiment 2, COCs (n = 581 were assigned to the same groups, but those in VS1, VS1 + VS2 and Vitrified groups were sub-divided and exposed to the warming solution for either 1 or 5 min. After treatment and/or warming, all COCs in both experiments underwent in vitro maturation, in vitro fertilization and in vitro culture. Results Cleavage and blastocyst rates did not differ among Control, VS1 and VS1 + VS2 groups in either experiment. In Experiment 2, there was no effect of time in the warming solution. However, both cleavage and blastocyst rates were lower (P  Conclusions The permeating cryoprotectants (EG and DMSO present in VS1 and VS2 solutions and the time in the warming solution containing sucrose had no adverse effects on cleavage and blastocyst rates of immature bovine COCs. However, cleavage rate and early embryo development were reduced following the vitrification and warming.

  7. Microtubule stabilisers docetaxel and paclitaxel reduce spindle damage and maintain the developmental competence of in vitro-mature bovine oocytes during vitrification.

    Science.gov (United States)

    Pitchayapipatkul, Jakkhaphan; Somfai, Tamás; Matoba, Satoko; Parnpai, Rangsan; Nagai, Takashi; Geshi, Masaya; Vongpralub, Thevin

    2017-09-01

    This study compared the efficacy of docetaxel (DT) and paclitaxel (PT) in reducing spindle damage during vitrification and maintaining the developmental competence of in vitro-matured (IVM) bovine oocytes after vitrification and warming. Pretreatment of IVM oocytes with 0.05µM DT for 30min before vitrification resulted in significantly higher (Pvitrification or those vitrified without pretreatment. When nuclear status and spindle morphology of vitrified oocytes were assess after warming by immunostaining, DT pretreatment before vitrification resulted in a significantly higher (Pbovine oocytes with 0.05µM DT or 1.0µM PT for 30min before vitrification reduces spindle damage to the same extent, without side effects on fertilisation and development. Pretreatment with 0.05µM DT improved the developmental competence of vitrified-warmed oocytes to a greater degree than 1.0µM PT pretreatment.

  8. Cement encapsulation of low-level waste liquids. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Baker, M.N.; Houston, H.M.

    1999-01-01

    Pretreatment of liquid high-level radioactive waste at the West Valley Demonstration Project (WVDP) was essential to ensuring the success of high-level waste (HLW) vitrification. By chemically separating the HLW from liquid waste, it was possible to achieve a significant reduction in the volume of HLW to be vitrified. In addition, pretreatment made it possible to remove sulfates, which posed several processing problems, from the HLW before vitrification took place.

  9. Effect of vitrification on the mRNA transcriptome of bovine oocytes.

    Science.gov (United States)

    Wang, N; Li, C-Y; Zhu, H-B; Hao, H-S; Wang, H-Y; Yan, C-L; Zhao, S-J; Du, W-H; Wang, D; Liu, Y; Pang, Y-W; Zhao, X-M

    2017-08-01

    Vitrification has been shown to decrease the developmental capacity of mammalian oocytes, and this is closely associated with the abnormal mRNA expressions of vitrified oocytes. However, the effect of vitrification on transcriptional machinery of oocytes examined by RNA sequencing (RNA-seq) has yet to be defined. In the present study, the mRNA transcriptomes of fresh and vitrified bovine oocytes were analysed by Smart-seq2 with the differently expressed genes determined by DEseq2 (an adjusted p-value of .05 and a minimum fold change of 2). The differentially expressed mRNAs were then searched against the Gene Ontology (GO) and Genomes (KEGG) database. Finally, the mRNA expressions of 10 candidate genes were validated using quantitative real-time PCR (qRT-PCR). Approximately 12,000 genes were detected in each sample of fresh or vitrified oocytes. Of these, the expression levels of 102 genes differed significantly in vitrified groups: 12 genes mainly involved in cell cycle, fertilization and glucose metabolism were upregulated, and 90 genes mainly involved in mitochondria, ribosomal protein, cytoskeleton, transmembrane protein, cell cycle and calcium ions were downregulated. GO analysis showed that these genes were mainly enriched in terms of membrane-bounded organelles, macromolecular complex, and intracellular part. The mRNA expression levels of 10 candidate genes selected randomly were in agreement with the results of the RNA-seq. In conclusion, our results showed that vitrification affected the mRNA transcriptome of bovine oocytes by downregulating genes, which contributed to the decreased developmental capacity of vitrified oocytes. Our findings will be useful in determining approaches to improve the efficiency of vitrified oocytes. © 2017 Blackwell Verlag GmbH.

  10. Vitrification of Rattus norvegicus immature cumulus-oocyte complexes using hyaluronic acid.

    Science.gov (United States)

    Paim, L M G; Gal, L L; Lopes, R F F; Oliveira, A T D

    2015-11-01

    The aim of this study was to assess in vitro meiosis resumption and nuclear maturation of Rattus norvegicus oocytes after vitrification with different cryoprotective solutions. Cumulus-oocyte complexes (COCs) were exposed to an equilibration solution for 4 min placed in cryoprotective solutions for 1 min and vitrified in open pulled straws. Cryoprotective solutions were prepared with 15% ethylene glycol + 15% dimethyl sulfoxide + 0.5 M sucrose and different supplements, to form the following groups: G1, 20% fetal bovine serum in modified phosphate-buffered saline (mPBS); G2, 0.4% bovine serum albumine in mPBS; G3, 1% hyaluronic acid in mPBS; and G4, 0.4% polyvinyl alcohol in mPBS. Seven days after vitrification, the COCs from G1 to G4 were warmed and in vitro matured for 30 h along with the control group. Hoechst staining was performed to assess meiosis resumption and nuclear maturation rates. Control group showed higher meiosis resumption (77.88%) and nuclear maturation rates (55.75%) compared to all vitrified groups. Among the vitrified COCs, G3 showed the highest meiosis resumption and nuclear maturation rates (G1, 26.5 and 15.38%; G2, 22.12 and 11.54%; G3, 34.55 and 20%; G4, 20.17 and 9.24%). Supplementation of the vitrification solution with 1% hyaluronic acid provided better results, compared to the other supplements. Hyaluronic acid can be useful to vitrify rat COCs associated with other cryoprotectant agents.

  11. Cryopreservation of bovine in vitro produced embryos using ethylene glycol in controlled freezing or vitrification.

    Science.gov (United States)

    Sommerfeld, V; Niemann, H

    1999-03-01

    In this study, the cryoprotectant ethylene glycol (EG) was tested for its ability to improve and facilitate the cryopreservation of in vitro produced (IVP) bovine embryos. Embryos were cryopreserved in EG solutions supplemented with either newborn calf serum (NBCS) or polyvinyl alcohol (PVA). To assess EG toxicity, the embryos were equilibrated in EG concentrations from 1.8 to 8.9 M at room temperature for 10 min and then cultured for 72 h on a cumulus cell monolayer. The hatching rate was highest for day 7 blastocysts frozen in 3.6 M EG (98%) and was not different from the control group (85%). The controlled freezing (0.3 degrees C/min to -35 degrees C) of expanded day 7 blastocysts resulted in a hatching rate of 81%, which was similar to that of the nonfrozen controls (76%). Differential staining revealed only very few degenerate blastomeres attributed to freezing and thawing. Upon direct nonsurgical transfer of day 7 expanded blastocysts frozen in 3.6 M EG, a pregnancy rate of 43% was achieved, while the pregnancy rate after transfer of other developmental stages was significantly lower (22% with expanded day 8 blastocysts). When bovine IVP embryos were incubated at room temperature in 7.2 M EG preceded by preequilibration in 3.6 M EG, the hatching rate of day 7 expanded blastocysts reached 93%. Upon vitrification of IVP day 7 and day 8 blastocysts and expanded blastocysts in 7.2 M EG, the latter showed a higher hatching rate (42%) than blastocysts (12%). Overall, PVA as supplement to the basic freezing solution instead of NBCS had deleterious effects on survival after controlled freezing or vitrification. The simple cryopreservation protocol employed in this study and the low toxicity of ethylene glycol highlight the usefulness of this approach for controlled freezing of IVP embryos. However, further experiments are needed to improve the pregnancy rate following embryo transfer and to enhance survival after vitrification. Copyright 1999 Academic Press.

  12. Vitrification of in vitro produced bovine embryos: in vitro and in vivo evaluations.

    Science.gov (United States)

    Martínez, A G; Valcárcel, A; de las Heras, M A; de Matos, D G; Furnus, C; Brogliatti, G

    2002-09-16

    The efficacy of different vitrification solutions to cryopreserve in vitro produced bovine blastocysts was evaluated based upon in vitro development of embryos in culture and on in vivo development of embryos transferred into recipients. In the first experiment, ethylene glycol + glycerol (Eg + Gly) + different sucrose concentrations were evaluated. There were no significant differences in development rates among solutions. As for hatching, the Eg + Gly + 0.1 M sucrose group had a greater rate as compared with Eg + Gly + 0 M sucrose and Eg + Gly + 0.5 M sucrose groups in the evaluations of Day 6, Day 7 and Day 6 + Day 7 embryos; and, Eg + Gly + 0.3 M sucrose group had a greater rate as compared with the Eg + Gly + 0 M sucrose and Eg + Gly + 0.5 M sucrose groups in evaluations of Day 6 and Day 6 + Day 7 embryos. There were no significant differences in development and hatching rates between Day 6 and 7 in in vitro produced bovine embryos within each treatment group. There were significant differences in nuclei number after vitrification between Eg + Gly + 0.1 M and Eg + Gly + 0 M sucrose groups and the Eg + Gly + 0.5 M sucrose group. Pregnancy after 60 days of transfer and calving rates showed a difference between in vivo produced embryos freshly transferred and in vitro produced embryos vitrified with Eg + Gly + 0.3 M. There were no significant differences in gestation length and sex ratio between treatments. As for birth weight, there were significant differences between fresh in vivo produced embryos and all treatments of in vitro produced embryos. There were significant differences in dystocial parturition between in vivo produced embryos and all treatments with in vitro produced embryos. These results demonstrate that vitrification can be used successfully in the cryopreservation of in vitro produced bovine embryos, and that it might be considered for use in commercial programs. Copyright 2002 Elsevier Science B.V.

  13. Analysis of the phospholipid profile of metaphase II mouse oocytes undergoing vitrification.

    Directory of Open Access Journals (Sweden)

    Jaehun Jung

    Full Text Available Oocyte freezing confers thermal and chemical stress upon the oolemma and various other intracellular structures due to the formation of ice crystals. The lipid profiles of oocytes and embryos are closely associated with both, the degrees of their membrane fluidity, as well as the degree of chilling and freezing injuries that may occur during cryopreservation. In spite of the importance of lipids in the process of cryopreservation, the phospholipid status in oocytes and embryos before and after freezing has not been investigated. In this study, we employed mass spectrometric analysis to examine if vitrification has an effect on the phospholipid profiles of mouse oocytes. Freshly prepared metaphase II mouse oocytes were vitrified using copper grids and stored in liquid nitrogen for 2 weeks. Fresh and vitrified-warmed oocytes were subjected to phospholipid extraction procedure. Mass spectrometric analyses revealed that multiple species of phospholipids are reduced in vitrified-warmed oocytes. LIFT analyses identified 31 underexpressed and 5 overexpressed phospholipids in vitrified mouse oocytes. The intensities of phosphatidylinositol (PI {18∶2/16∶0} [M-H]- and phosphatidylglycerol (PG {14∶0/18∶2} [M-H]- were decreased the most with fold changes of 30.5 and 19.1 in negative ion mode, respectively. Several sphingomyelins (SM including SM {d38∶3} [M+H]+ and SM {d34∶0} [M+K]+ were decreased significantly in positive ion mode. Overall, the declining trend of multiple phospholipids demonstrates that vitrification has a marked effect on phospholipid profiles of oocytes. These results show that the identified phospholipids can be used as potential biomarkers of oocyte undergoing vitrification and will allow for the development of strategies to preserve phospholipids during oocyte cryopreservation.

  14. Evaluation of alternative chemical additives for high-level waste vitrification feed preparation processing

    Energy Technology Data Exchange (ETDEWEB)

    Seymour, R.G.

    1995-06-07

    During the development of the feed processing flowsheet for the Defense Waste Processing Facility (DWPF) at the Savannah River Site (SRS), research had shown that use of formic acid (HCOOH) could accomplish several processing objectives with one chemical addition. These objectives included the decomposition of tetraphenylborate, chemical reduction of mercury, production of acceptable rheological properties in the feed slurry, and controlling the oxidation state of the glass melt pool. However, the DEPF research had not shown that some vitrification slurry feeds had a tendency to evolve hydrogen (H{sub 2}) and ammonia (NH{sub 3}) as the result of catalytic decomposition of CHOOH with noble metals (rhodium, ruthenium, palladium) in the feed. Testing conducted at Pacific Northwest Laboratory and later at the Savannah River Technical Center showed that the H{sub 2} and NH{sub 3} could evolve at appreciable rates and quantities. The explosive nature of H{sub 2} and NH{sub 3} (as ammonium nitrate) warranted significant mitigation control and redesign of both facilities. At the time the explosive gas evolution was discovered, the DWPF was already under construction and an immediate hardware fix in tandem with flowsheet changes was necessary. However, the Hanford Waste Vitrification Plant (HWVP) was in the design phase and could afford to take time to investigate flowsheet manipulations that could solve the problem, rather than a hardware fix. Thus, the HWVP began to investigate alternatives to using HCOOH in the vitrification process. This document describes the selection, evaluation criteria, and strategy used to evaluate the performance of the alternative chemical additives to CHOOH. The status of the evaluation is also discussed.

  15. Experimental design for the optimization of propidium monoazide treatment to quantify viable and non-viable bacteria in piggery effluents.

    Science.gov (United States)

    Desneux, Jérémy; Chemaly, Marianne; Pourcher, Anne-Marie

    2015-08-16

    Distinguishing between viable and dead bacteria in animal and urban effluents is a major challenge. Among existing methods, propidium monoazide (PMA)-qPCR is a promising way to quantify viable cells. However, its efficiency depends on the composition of the effluent, particularly on total suspended solids (TSS)) and on methodological parameters. The aim of this study was evaluate the influence of three methodological factors (concentration of PMA, incubation time and photoactivation time) on the efficiency of PMA-qPCR to quantify viable and dead cells of Listeria monocytogenes used as a microorganism model, in two piggery effluents (manure and lagoon effluent containing 20 and 0.4 TSS g.kg(-1), respectively). An experimental design strategy (Doehlert design and desirability function) was used to identify the experimental conditions to achieve optimal PMA-qPCR results. The quantification of viable cells of L. monocytogenes was mainly influenced by the concentration of PMA in the manure and by the duration of photoactivation in the lagoon effluent. Optimal values differed with the matrix: 55 μM PMA, 5 min incubation and 56 min photoactivation for manure and 20 μM PMA, 20 min incubation and 30 min photoactivation for lagoon effluent. Applied to five manure and four lagoon samples, these conditions resulted in satisfactory quantification of viable and dead cells. PMA-qPCR can be used on undiluted turbid effluent with high levels of TSS, provided preliminary tests are performed to identify the optimal conditions.

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

    Science.gov (United States)

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

    2017-01-01

    Cryopreservation is known for its marked deleterious effects on embryonic health. Bovine compact morulae were vitrified or slow-frozen, and post-warm morulae were cultured to the expanded blastocyst stage. Blastocysts developed from vitrified and slow-frozen morulae were subjected to microarray analysis and compared with blastocysts developed from unfrozen control morulae for differential gene expression. Morula to blastocyst conversion rate was higher (P vitrification but those which survived up to blastocyst stage did not express ≥ 2 fold change in their gene expression as compared with blastocysts from control morulae.

  17. In vitro assessment of a direct transfer vitrification procedure for bovine embryos.

    Science.gov (United States)

    Campos-Chillòn, L F; Walker, D J; de la Torre-Sanchez, J F; Seidel, G E

    2006-04-01

    We developed a simple vitrification technique for bovine embryos that could permit direct transfer. Embryos were produced in-vitro by standard procedures. The base medium for cryopreservation was a chemically defined medium similar to SOF + 25 mM Hepes and 0.25% fatty acid free bovine serum albumin (FAF-BSA) (HCDM2). In experiment 1, embryos were first exposed to 3.5M ethylene glycol (V1) for 1, 2 or 3 min at room temperature (20-24 degrees C), and then moved to 7 M ethylene glycol (V2) at 4 or 20-24 degrees C and loaded in 0.25-mL straws. After 45 s in 7 M ethylene glycol, straws were placed in liquid nitrogen. Embryos that were loaded at 20-24 degrees C had higher survival rates than those loaded at 4 degrees C (Pembryos survived well after 15 min in straws if warmed at 37 degrees C. In experiment 3, ethylene glycol concentration (3, 4 or 5 M) and exposure time (0.5 or 1 min) during two-step addition of cryoprotectant were studied for bovine morulae. In experiment 4, morulae were exposed to V2 for 30 or 45 s in HCDM2 or Vigro holding medium and then held in 22-24 degrees C air or 37 degrees C water post-warming. Experiment 5 was like experiment 4 except blastocysts were used. Overall survival rates of blastocysts in experiment 5 averaged 80% of non-vitrified controls after 48 h culture. The survival rates with in vitro-produced morulae in experiments 1, 3 and 4 were unacceptable. Vitrification solutions based on Vigro tended to result in higher survival than HCDM2 for blastocysts, but not morulae. In experiment 6, the survival rate in vitro of in vivo-produced morulae and blastocysts after two-step vitrification was nearly 100%. Our vitrification technique was very effective for in vitro produced blastocysts, but not for in vitro-produced morulae.

  18. Hanford Waste Vitrification Plant full-scale feed preparation testing with water and process simulant slurries

    Energy Technology Data Exchange (ETDEWEB)

    Gaskill, J.R.; Larson, D.E.; Abrigo, G.P. [and others

    1996-03-01

    The Hanford Waste Vitrification Plant was intended to convert selected, pretreated defense high-level waste and transuranic waste from the Hanford Site into a borosilicate glass. A full-scale testing program was conducted with nonradioactive waste simulants to develop information for process and equipment design of the feed-preparation system. The equipment systems tested included the Slurry Receipt and Adjustment Tank, Slurry Mix Evaporator, and Melter-Feed Tank. The areas of data generation included heat transfer (boiling, heating, and cooling), slurry mixing, slurry pumping and transport, slurry sampling, and process chemistry. 13 refs., 129 figs., 68 tabs.

  19. Bulk Vitrification Performance Enhancement: Refractory Lining Protection Against Molten Salt Penetration

    Energy Technology Data Exchange (ETDEWEB)

    Hrma, Pavel R.; Bagaasen, Larry M.; Schweiger, Michael J.; Evans, Michael B.; Smith, Benjamin T.; Arrigoni, Benjamin M.; Kim, Dong-Sang; Rodriguez, Carmen P.; Yokuda, Satoru T.; Matyas, Josef; Buchmiller, William C.; Gallegos, Autumn B.; Fluegel, Alexander

    2007-08-06

    Bulk vitrification (BV) is a process that heats a feed material that consists of glass-forming solids and dried low-activity waste (LAW) in a disposable refractory-lined metal box using electrical power supplied through carbon electrodes. The feed is heated to the point that the LAW decomposes and combines with the solids to generate a vitreous waste form. This study supports the BV design and operations by exploring various methods aimed at reducing the quantities of soluble Tc in the castable refractory block portion of the refractory lining, which limits the effectiveness of the final waste form.

  20. CRYOPRESERVATION OF SWEET POTATO SHOOT TIPS USING A DROPLET-VITRIFICATION PROCEDURE.

    Science.gov (United States)

    Park, S U; Kim, H H

    2015-01-01

    Sweet potato is a staple food worldwide, but a problematic species in terms of long term storage, as it is not suitable for germplasm conservation. This study aimed to develop cryopreservation protocols for sweet potato shoot tips based on a droplet-vitrification procedure. As a standard procedure, sweet potato shoot tips were precultured in a liquid MS medium supplemented with 10% sucrose (S-10%) and 17.5% sucrose (S-17.5%) for 31 and 17 h, respectively. They were then osmoprotected with C4-35% (17.5% glycerol + 17.5% sucrose) for 50 min and cryoprotected with PVS3 (50% glycerol + 50% sucrose) for 60 min. A set of experiments was designed to investigate critical factors, i.e. stepwise sucrose preculture, osmoprotection, cryoprotection with PVS2- and PVS3-based vitrification solutions, and their combinational effect, as well as temperature alteration through placement in a cooling/rewarming container. Sucrose preculture was determined to be necessary for the adaptation of sweet potato shoot tips to cryoprotection with PVS3, and the highest post-thaw (LN) regeneration rate was observed in a preculture with S-10% for 31 h → S-17.5% for 17 h (19.0%). The effect of one-step or two-step osmoprotection was not significant on survival or regeneration of either the cryoprotected-control (LNC) or LN shoot tips. Responses of sweet potato shoot tips to osmoprotection and cryoprotection were linked to the level of sucrose preculture. The use of alumimium foil strips (droplet-vitrification) resulted in significantly higher LN survival (89.8%) and regeneration (19.0%), compared to those using cryovials (vitrification, 67.2% and 0%, respectively). LN regeneration increased by 67.5% when cryopreserved shoot tips were transferred to a new postculture medium. This study demonstrates that the combination of stepwise sucrose preculture with a higher final concentration (up to 17.5%), cryoprotection with PVS3 and cooling with foil strip is crucial to the regeneration of LN sweet

  1. Hanford Waste Vitrification Plant technical background document for best available radionuclide control technology demonstration

    Energy Technology Data Exchange (ETDEWEB)

    Carpenter, A.B.; Skone, S.S.; Rodenhizer, D.G.; Marusich, M.V. (Ebasco Services, Inc., Bellevue, WA (USA))

    1990-10-01

    This report provides the background documentation to support applications for approval to construct and operate new radionuclide emission sources at the Hanford Waste Vitrification Plant (HWVP) near Richland, Washington. The HWVP is required to obtain permits under federal and state statutes for atmospheric discharges of radionuclides. Since these permits must be issued prior to construction of the facility, draft permit applications are being prepared, as well as documentation to support these permits. This report addresses the applicable requirements and demonstrates that the preferred design meets energy, environmental, and economic criteria for Best Available Radionuclide Control Technology (BARCT) at HWVP. 22 refs., 11 figs., 25 tabs.

  2. Literature review of arc/plasma, combustion, and joule-heated melter vitrification systems

    Energy Technology Data Exchange (ETDEWEB)

    Freeman, C.J.; Abrigo, G.P.; Shafer, P.J.; Merrill, R.A.

    1995-07-01

    This report provides reviews of papers and reports for three basic categories of melters: arc/plasma-heated melters, combustion-heated melters, and joule-heated melters. The literature reviewed here represents those publications which may lend insight to phase I testing of low-level waste vitrification being performed at the Hanford Site in FY 1995. For each melter category, information from those papers and reports containing enough information to determine steady-state mass balance data is tabulated at the end of each section. The tables show the composition of the feed processed, the off-gas measured via decontamination factors, gross energy consumptions, and processing rates, among other data.

  3. The search for viable local government system in Nigeria: an ...

    African Journals Online (AJOL)

    The history of the Nigerian local government system has been one long episode of trails and errors aimed at achieving viable local government institution without much success. Local government in the country began its long series of reforms from the colonial period when the colonial government attempted to ...

  4. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    A rapid and sensitive assay was developed for the detection of low numbers of viable Vibrio cholerae and Shigella spp. cells in environmental and drinking water samples. Water samples were filtered, and the filters were enriched in a non-selective medium. The enrichment cultures were prepared for polymerase chain ...

  5. Comment: Towards a Viable Local Government Structure in Nigeria ...

    African Journals Online (AJOL)

    Local governments are principally established for development at the grassroots and they must be structured in a manner that makes them viable and capable of achieving this purpose. The objective of this comment is to appraise the current local government structure under the Nigerian constitutional framework with a view ...

  6. Cultivation and multiplication of viable axenic Trypanosoma vivax in ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-01

    Sep 1, 2009 ... Cultivation and multiplication of viable axenic. Trypanosoma vivax in vitro and in vivo. O. A. Idowu, A. B. Idowu, C. F. Mafiana and S. O. Sam-Wobo*. Parasitology Laboratory, Department of Biological Sciences, University of Agriculture, Abeokuta, Nigeria. Accepted 13 April, 2006. Trypanosoma vivax was ...

  7. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    DRINIE

    2003-04-02

    Apr 2, 2003 ... A rapid and sensitive assay was developed for the detection of low numbers of viable Vibrio cholerae and Shigella spp. cells in environmental and drinking water samples. Water samples were filtered, and the filters were enriched in a non-selective medium. The enrichment cultures were prepared for ...

  8. High speed flow cytometric separation of viable cells

    Science.gov (United States)

    Sasaki, Dennis T.; Van den Engh, Gerrit J.; Buckie, Anne-Marie

    1995-01-01

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  9. Vitrification of in vitro produced bovine embryos: effect of embryonic block and developmental kinetics.

    Science.gov (United States)

    Asgari, V; Hosseini, S M; Forouzanfar, M; Hajian, M; Nasr-Esfahani, M H

    2012-12-01

    In order to investigate whether the kinetics and stage of embryo development affect cryosurvival of in vitro produced bovine embryos, cleaved embryos were categorized in six groups based on their developmental kinetics regarding the stage of embryonic block in bovine (8-16 cell stage): I and II--early (day 2) and late (day 3) 5-8 cell, III and IV--early (day 3) and late (day 4) 8-16 cell, and V and VI--early (day 4) and late (day 5) morula. The cryosurvival and developmental competence of these embryos were compared with each other and also with the corresponding control groups. The potential of 5-8 cell stage embryos to survive vitrification and further develop towards blastocyst stage was significantly lower than vitrified and un-vitrified 8-16 cell and morula stage embryos. These results suggest that, the survival rate and potential of embryos to develop towards blastocyst stage might be affected by the kinetic of the embryo development. Moreover, the results of this study indicated that the optimal stages of early embryo vitrification are post-embryonic block. Copyright © 2012 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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.

  11. Bovine oocytes in secondary follicles grow in medium containing bovine plasma after vitrification.

    Science.gov (United States)

    Taketsuru, Hiroaki; Takajo, Asuka; Bao, Rong-Mei; Hamawaki, Atsushi; Yoshikawa, Motoichi; Miyano, Takashi

    2011-02-01

    There has been no culture system that supports the growth of bovine oocytes for more than 2 weeks. In the present study, bovine secondary follicles were cultured for 4 weeks, and the effects of supplemented protein components and FSH in the culture medium on the growth of the oocytes were examined. The effect of vitrification of secondary follicles on the subsequent oocyte growth was also examined. Secondary follicles (150 to 200 µm in diameter) containing growing oocytes (approximately 60 µm in diameter) were dissected from ovaries and cultured in a medium supplemented with FSH (0, 25 or 50 ng/ml) and one of the following four kinds of protein components: bovine serum albumin (BSA), bovine plasma (BPL), fetal calf serum (FCS) and bovine follicular fluid (BFF). In BSA- and BPL-supplemented media with 0 or 25 ng/ml FSH, more than 50% of follicles showed no degenerative signs during culture, and oocytes significantly increased in size after 4 weeks (Pbovine secondary follicles for 4 weeks, even after vitrification and warming of the follicles.

  12. Effect of antifreeze glycoprotein 8 supplementation during vitrification on the developmental competence of bovine oocytes.

    Science.gov (United States)

    Liang, Shuang; Yuan, Bao; Kwon, Jeong-Woo; Ahn, Mija; Cui, Xiang-Shun; Bang, Jeong Kyu; Kim, Nam-Hyung

    2016-07-15

    The purpose of this study was to investigate the effect of antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification on the survival, fertilization, and embryonic development of bovine oocytes and the underlying molecular mechanism(s). Survival, fertilization, early embryonic development, apoptosis, DNA double-strand breaks, reactive oxygen species levels, meiotic cytoskeleton assembly, chromosome alignment, and energy status of mitochondria were measured in the present experiments. Compared with that in the nonsupplemented group; survival, monospermy, blastocyst formation rates, and blastomere counts were significantly higher in the AFGP8-supplemented animals. Oocytes of the latter group also presented fewer double-strand breaks and lower cathepsin B and caspase activities. Rates of normal spindle organization and chromosome alignment, actin filament impairment, and mitochondrial distribution were significantly higher in the AFGP8-supplemented group. In addition, intracellular reactive oxygen species levels significantly decreased in the AFGP8-supplemented groups, maintaining a higher ΔΨm than that in the nonsupplemented group. Taken together, these results indicated that supplementation with AFGP8 during vitrification has a protective effect on bovine oocytes against chilling injury. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Vitrification of bovine blastocysts produced in vitro inflicts selective damage to the inner cell mass.

    Science.gov (United States)

    Gómez, E; Muñoz, M; Rodríguez, A; Caamaño, J N; Facal, N; Díez, C

    2009-04-01

    In contrast to the embryos derived from live animals, the embryos produced in vitro undergo increased damage and reduced survival after cryopreservation, particularly when produced with serum. In medium containing serum, retinoic acid increases cell numbers in the inner cell mass and the trophectoderm without altering their relative proportions in the bovine blastocyst. In this work, in medium without serum, we analyzed the contribution of retinoic acid to the development of blastocyst and survival to vitrification, and found a strong cell reduction in the inner mass when compared to the trophectoderm. Day-6 in vitro-produced morulae were treated for 24 h with retinoic acid (0.7 and 1.4 microm) and subsequently cultured without additives for a further 24 h period. Day-8 blastocyst production and cell counts in hatched blastocysts were unaffected by retinoic acid. However, Day-7 expanded, vitrified embryos produced with retinoic acid 1.4 microm survived at lower rates than controls when cultured after warming. Vitrification greatly reduced cell numbers in the inner mass (p vitro survival to cryopreservation is sometimes scarcely informative on the viability of the embryo after transfer to recipients.

  14. High survival and hatching rates following vitrification of embryos at blastocyst stage: a bovine model study.

    Science.gov (United States)

    Huang, Jack Y J; Chung, Jin-Tae; Tan, Seang Lin; Chian, Ri-Cheng

    2007-04-01

    Cryopreservation of embryos at the blastocyst stage may provide an effective method to increase the cumulative pregnancy rate for each treatment cycle of ovarian-stimulated IVF. The objective of this study was to evaluate the survival rate and hatching rate of bovine blastocysts following vitrification using a method designed for oocytes, with a view to introducing this methodology into human assisted reproduction technology and reproductive medicine. Bovine blastocysts were produced from abattoir materials subjected to in-vitro maturation and in-vitro fertilization. Survival rate of the bovine blastocysts was 100% (94/94) following vitrification using a method designed for oocyte cryopreservation. There was no difference in the hatching rate of the bovine blastocysts between control (62.5%: 60/96) and vitrified (61.7%: 58/94) groups. The number of dead cells in the blastocysts was not significantly different between control (5.0 +/- 2.9) and vitrified (9.5 +/- 4.0) groups. In conclusion, the results of this study indicate that bovine blastocysts can be vitrified successfully using a procedure designed for oocyte cryopreservation. It is possible that this method may also be successful for the cryopreservation of human embryos. A further study into this is currently being organized.

  15. Behavior of lateral buds of Hancornia speciosa after cryopreservation by encapsulation-vitrification

    Directory of Open Access Journals (Sweden)

    Débora de Oliveira Prudente

    2017-05-01

    Full Text Available Hancornia speciosa is a fruitful species from Cerrado biome with high economic potential. However, the intense and disordered extractivism have caused a reduction of its population in its endemic area. In addition, seed recalcitrance negatively affects the conventional conservation of the species. Aiming to find alternatives that enable the long-term conservation of this species, the study’s objective was to assess the behavior of lateral bud’s regrowth after cryopreservation procedures by encapsulation-vitrification technique. Sodium alginate capsules containing lateral buds were pre-cultured in liquid WPM supplemented with 1.0 M glycerol, and subsequently exposed to different concentrations of sucrose (0.3; 0.75 and 1.0 M for 24 or 48 hours. The capsules were subjected to dehydration in silica gel or airflow hood for 0, 1, 2 and 3 hours before different incubation times in PVS2 (0, 15, 30, 60 and 120 minutes at 0°C. A high regeneration percentage of lateral buds was observed after cryopreservation of capsules treated with 0.75 M sucrose plus 1.0 M glycerol (24 hours, associated with dehydration in an airflow hood (1 hour and immersion in PVS2 (15 minutes. Encapsulation-vitrification allowed the long-term conservation, and provided high plant material survival rates after cryopreservation of Hancornia speciosa sensitive explants.

  16. Ultrastructure of preimplantation genetic diagnosis-derived human blastocysts grown in a coculture system after vitrification.

    Science.gov (United States)

    Escribá, María-José; Escobedo-Lucea, Carmen; Mercader, Amparo; de los Santos, María-José; Pellicer, Antonio; Remohí, José

    2006-09-01

    To evaluate ultrastructural features of preimplantation genetic diagnosis (PGD) blastocysts before and after vitrification. Descriptive study of both vitrified and fresh hatching blastocysts. PGD program at the Instituto Universitario, Instituto Valenciano de Infertilidad. Patients undergoing PGD donated their abnormal embryos for research (n = 26). Biopsied embryos were cultured in the presence of human endometrial cells until day 6. Sixteen blastocysts were vitrified. A total of 11 high-scored hatching blastocysts, 6 warmed and 5 fresh, were fixed for ultrastructure. The cytoskeleton structure, type of intercellular junctions, and basic intracellular organelles in trophoectoderm cells and the inner cell mass were analyzed. Ten of 16 blastocysts (62%) survived the warming process. Six of these showed no signs of cell degeneration and light microscopy revealed similar ultrastructural characteristics to those of controls. However, in trophoectoderm cells from both fresh and cryopreserved blastocysts, a reduced number of tight junctions and the presence of degradation bodies were detected. The particular ultrastructural features observed in PGD-derived blastocysts could be related to embryo manipulation and culture conditions. Vitrification does not seem to alter blastocysts, as those that survive hatching do not display detectable cellular alterations when observed through electron microscopy.

  17. Cold-cap reactions in vitrification of nuclear waste glass: Experiments and modeling

    Energy Technology Data Exchange (ETDEWEB)

    Chun, Jaehun; Pierce, David A. [Pacific Northwest National Laboratory, Richland, WA 99352 (United States); Pokorný, Richard [Department of Chemical Engineering, Institute of Chemical Technology in Prague, Technická 5, 166 28 Prague 6 (Czech Republic); Hrma, Pavel, E-mail: pavelhrma@postech.ac.kr [Pacific Northwest National Laboratory, Richland, WA 99352 (United States); Division of Advanced Nuclear Engineering, Pohang University of Science and Technology, Pohang (Korea, Republic of)

    2013-05-10

    Highlights: ► We measured enthalpy-based conversion degree of overlapping reactions using DSC. ► We employed the run/rerun technique to obtain heat flow associated with reactions. ► Batch-to-glass conversion advances via multiple overlapping reactions. ► The kinetic model is intended for the source term in the energy transfer equation. ► The results are relevant for industrial glass making and nuclear waste vitrification. - Abstract: Cold-cap reactions are multiple overlapping reactions that occur in the waste-glass melter during the vitrification process when the melter feed is being converted to molten glass. In this study, we used simultaneous differential scanning calorimetry–thermogravimetry (DSC–TGA) to investigate cold-cap reactions in a high-alumina high-level waste melter feed. To separate the reaction heat from both the heat associated with the heat capacity of the feed and experimental artifacts, we employed the run/rerun method, which enabled us to define the degree of conversion based on the reaction heat and to estimate the heat capacity of the reacting feed. Assuming that the reactions are nearly independent and can be approximated by an nth order kinetic model, we obtained the kinetic parameters using the Kissinger method combined with least squares analysis. The resulting mathematical simulation of the cold-cap reactions provides a key element for the development of an advanced cold-cap model.

  18. How thermal stress alters the confinement of polymers vitrificated in nanopores

    Science.gov (United States)

    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.

  19. Development of a vitrification-based cryopreservation protocol for the storage of saltcedar (Tamarix boveana Bunge).

    Science.gov (United States)

    Cano-Castillo, M; Casas, J L

    2012-01-01

    We cryopreserved in vitro shoot tips of saltcedar (Tamarix boveana Bunge) using the vitrification technique. The success of the cryopreservation protocol was strongly affected by preculture, loading duration, dehydration duration in plant vitrification solution 2 (PVS2), and medium composition during post-warming regrowth. The highest explant regrowth (50 percent) occurred when the following conditions were employed: preculture in 0.4 M glycerol; treatment with a loading solution (LS) consisting of 2 M glycerol + 0.4 M sucrose in culture medium for 40 min at room temperature; and dehydration in PVS2 at 0 degree C for 45 min before rapid immersion in liquid nitrogen (LN). Rewarming was performed in a water-bath at 40 degree C for 2 min. Explants were then immersed in unloading solution for 10 min before plating on recovery medium supplemented with 0.01 mg per liter thidiazuron (TDZ). TDZ was progressively eliminated from the medium over a period of 6 weeks. Plantlets were transferred to a double-layer medium to enhance rooting. This protocol was successfully applied to three individuals of T. boveana harvested from the wild.

  20. Transportable Vitrification System RCRA Closure Practical Waste Disposition Saves Time And Money

    Energy Technology Data Exchange (ETDEWEB)

    Brill, Angie; Boles, Roger; Byars, Woody

    2003-02-26

    The Transportable Vitrification System (TVS) was a large-scale vitrification system for the treatment of mixed wastes. The wastes contained both hazardous and radioactive materials in the form of sludge, soil, and ash. The TVS was developed to be moved to various United States Department of Energy (DOE) facilities to vitrify mixed waste as needed. The TVS consists of four primary modules: (1) Waste and Additive Materials Processing Module; (2) Melter Module; (3) Emissions Control Module; and (4) Control and Services Module. The TVS was demonstrated at the East Tennessee Technology Park (ETTP) during September and October of 1997. During this period, approximately 16,000 pounds of actual mixed waste was processed, producing over 17,000 pounds of glass. After the demonstration was complete it was determined that it was more expensive to use the TVS unit to treat and dispose of mixed waste than to direct bury this waste in Utah permitted facility. Thus, DOE had to perform a Resource Conservation and Recovery Act (RCRA) closure of the facility and find a reuse for as much of the equipment as possible. This paper will focus on the following items associated with this successful RCRA closure project: TVS site closure design and implementation; characterization activities focused on waste disposition; pollution prevention through reuse; waste minimization efforts to reduce mixed waste to be disposed; and lessons learned that would be integrated in future projects of this magnitude.

  1. Solid-surface vitrification and in-straw dilution after warming of in vitro-produced bovine embryos.

    Science.gov (United States)

    Rodriguez-Villamil, P; Ongaratto, F L; Fernandez Taranco, M; Bó, G A

    2014-02-01

    Three experiments were designed to test a solid-surface vitrification system for bovine in vitro-produced embryos and to develop a simple method of in-straw dilution after warming, which can be potentially used for direct transfer in the field. Experiment 1 evaluated embryo survival rates (i.e. re-expansion and hatching) after vitrification and warming in three different solutions: VS1 (20% ethylene glycol (EG) + 20% propanediol (PROH) + 0.25 m trehalose (Tr)), VS2 (20% EG + 1M Tr) or VS3 (30% EG + 0.75 m Tr). Re-expansion and hatching rates were higher (p vitrification: glass micropipettes or solid surface, using the VS1 or VS3 solutions. No significant differences were detected between the two methods; but re-expansion and hatching rates were higher (p vitrification using simplified EG-based solutions and in-straw dilution with holding media may be a practical alternative for cryopreservation and direct transfer of in vitro-produced bovine embryos. © 2013 Blackwell Verlag GmbH.

  2. Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification.

    Science.gov (United States)

    Sudano, Mateus José; Paschoal, Daniela Martins; Rascado, Tatiana da Silva; Magalhães, Luis Carlos Oña; Crocomo, Letícia Ferrari; de Lima-Neto, João Ferreira; Landim-Alvarenga, Fernanda da Cruz

    2011-04-15

    The objective was to evaluate supplementation of fetal calf serum (FCS) and phenazine ethosulfate (PES), a metabolic regulator that inhibits fatty acid synthesis, in culture media during in vitro production (IVP) of bovine embryos. Taking oocyte fertilization (n = 4,320) as Day 0, four concentrations of FCS (0, 2.5, 5, and 10%) and three periods of exposure to PES (without addition-CONTROL; after 60 h-PES Day 2.5 of embryo culture; and after 96 h-PES Day 4) were evaluated. Increasing FCS concentration in the culture media enhanced lipid accumulation (P vitrification (2.5%: 81.6 ± 2.5 vs 10%: 67.3 ± 3.5, P vitrification ( 72.0 ± 3.0 vs PES Day 2.5: 79.9 ± 2.8 or PES Day 4: 86.2 ± 2.4, P vitrification. Moreover, embryo quality, represented by the fresh apoptosis rate, was better than lipid content for predicting embryo survival after vitrification. Copyright © 2011 Elsevier Inc. All rights reserved.

  3. Application of vitrification-derived cryotechniques for long-term storage of poplar and aspen (Populus spp. germplasm

    Directory of Open Access Journals (Sweden)

    I. TSVETKOV

    2008-12-01

    Full Text Available The application of three different vitrification-based freezing strategies for the cryostorage of white poplar (Populus alba L. and hybrid aspen (P. tremula L. × P. tremuloides Michx. have been assessed. The PVS2 vitrification protocol was successfully applied to two white poplar in vitro clones stored for more than 6 months in slow-growth conditions (4 °C, in darkness and showing clear signs of explant etiolation and decay. After 60 min of PVS2 treatment, P. alba L. (cv. Villafranca explants isolated from axillary buds demonstrated significantly better potential for post-freeze regrowth (64% compared to those obtained from apical buds (17%. Similarly, a high level of survival (78% of the frozen hybrid aspen shoot tips was recorded following the application of the same technique. Using the ‘encapsulation-vitrification’ procedure, no toxic effects of the PVS2 treatment were noticed after 120 min exposure, however none of the cryopreserved (poplar and aspen explants survived after 3 weeks. In contrast, the ‘droplet-vitrification’ technique appeared to be very efficient in the cryopreservation of white poplar shoot tips, which increases the opportunities for wider application of this method in other woody species.;

  4. Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

    Directory of Open Access Journals (Sweden)

    Alane Pains Oliveira do Monte

    2015-12-01

    Full Text Available This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced in vivo. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect or not (Direct. The embryo quality was classified as embryos of degrees I (excellent or good, II (regular, III (poor and IV (dead or degenerate. It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P < 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%, higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 % while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.

  5. Cryopreservation of human failed-matured oocytes followed by in vitro maturation: vitrification is superior to the slow freezing method

    Directory of Open Access Journals (Sweden)

    Zhang ZhiGuo

    2011-12-01

    Full Text Available Abstract Background Oocyte cryopreservation is an important method used in a number of human fertility circumstances. Here, we compared the survival, in vitro maturation, fertilization, and early embryonic development rates of frozen-thawed human immature oocytes using two different cryopreservation methods. Methods A total of 454 failed-matured oocytes [germinal vesicle (GV and metaphase I (MI stages] were collected from 135 patients (mean age 33.84 +/- 5.0 y who underwent intracytoplasmic sperm injection (ICSI cycles between February 2009 and December 2009 and randomly divided into a slow freezing group [1.5 mol/L-1, 2-propanediol (PROH + 0.2 mol/l sucrose] and vitrification group [20% PROH + 20% ethylene glycol (EG + 0.5 mol/l sucrose]. Results The vitrification protocol yielded a better survival rate than the slow freezing protocol at each maturation stage assessed. Regardless of the maturation stage (GV + MI, the slow freezing protocol had a significantly lower survival rate than the vitrification protocol (p in vitro maturation (21.2 vs. 54.0%, respectively; p 0.05. For the GV-matured oocytes, no fertilized eggs were obtained in the slow-freezing group, while a 19.0% (4/21 fertilization rate was observed in the vitrification group. For the MI-matured oocytes, fertilization rates for the slow freezing and vitrified groups were 36% and 61.1%, respectively, but no significant difference was found between the two groups (PIn the Methods section in the MS, all procedures were compliant with ethical guidelines, i.e. approved by the Ethical Committee of our university and Informed Consent signed by each patient. > 0.05. In the GV vitrification group, no embryo formed; however, in the MI slow freezing group, 12 oocytes were fertilized, but only two achieved cleavage and were subsequently blocked at the 2-cell stage. In the MI vitrification group, a total of 22 embryos were obtained, five of which developed to the blastocyst stage. Conclusions

  6. Removal of viable bacteria and endotoxins by Electro Deionization (EDI).

    Science.gov (United States)

    Harada, Norimitsu; Otomo, Teruo; Watabe, Tomoichi; Ase, Tomonobu; Takemura, Takuto; Sato, Toshio

    2011-09-01

    Viable bacteria and endotoxins in water sometimes cause problems for human health. Endotoxins are major components of the outer cell wall of gram-negative bacteria (lipopolysaccharides). In medical procedures, especially haemodialysis (HD) and related therapies (haemodiafiltration (HDF), haemofiltration (HF)), endotoxins in the water for haemodialysis can permeate through the haemodialysis membrane and cause microinflammation or various haemodialysis-related illnesses. To decrease such a biological risk, RO and UF membranes are generally used. Also, hot water disinfection or the chemical disinfection is regularly executed to kill bacteria which produce endotoxins. However, simple treatment methods and equipment may be able to decrease the biological risk more efficiently. In our experiments, we confirmed that viable bacteria and endotoxins were removed by Electro Deionization (EDI) technology and also clarified the desorption mechanisms.

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

    Directory of Open Access Journals (Sweden)

    Wan-Hafizah Wan Jusof

    2015-07-01

    Full Text Available 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 developmental viability compared to those that cleaved at a later period (LC. However, the viability 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 oviducts were excised and embryos harvested. Embryos at the 2-cell stage were categorized 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% CO2 atmosphere. Control embryos were cultured until the blastocyst stage without vitrification. Experimental embryos at the 2-cell stage were vitrified 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. Preselection of EC embryos may be used as a tool for selection of embryos that exhibit higher developmental competence after vitrification.

  8. Meiotic maturation and developmental capability of ovine oocytes at germinal vesicle stage following vitrification using different cryodevices.

    Science.gov (United States)

    Quan, Guo Bo; Wu, Guo Quan; Wang, Ya Jing; Ma, Yuan; Lv, Chun Rong; Hong, Qiong Hua

    2016-02-01

    In order to assess effects of vitrification on ovine oocytes at the germinal vesicle (GV) stage, the conventional plastic straw (CS), the open-pulled straw (OPS), and Cryoloop were used to vitrify ovine oocytes. Oocytes were randomly divided into five groups: (1) Control; (2) Oocytes exposed to vitrification and dilution solutions without any cryopreservation (toxicity); (3) Oocytes vitrified using CS (CS); (4) Oocytes vitrified using OPS (OPS), and (5) Oocytes vitrified using Cryoloop (Cryoloop). The viability, cumulus cell expansion, nuclear maturation after in vitro maturation (IVM), and developmental capability of vitrified oocytes following parthenogenetic activation (PA) or in vitro fertilization (IVF) were assessed. The pretreatment in the vitrification and dilution solutions without any freezing or thawing did not adversely influence oocytes. The viability of vitrified oocytes were significantly declined compared to unfrozen oocytes (P straws or Cryoloop was significantly higher than that in the CS group (P plastic straws was significantly less than those of the other freezing groups (P straws. However, the cleavage rate of vitrified oocytes in the CS group was significantly less than that in the OPS or Cryoloop group (P plastic straw developed to the blastocyst stage following IVF. There was no significant difference existing between OPS and Cryoloop with respect to the blastocyst rate. After staining with cFDA and PI, cumulus cells surrounding oocytes were partly damaged by vitrification and thawing while the membrane of vitrified oocyte still remained intact. In conclusion, vitrification can seriously damage ovine immature oocytes and cumulus cells surrounding oocytes, which may subsequently affect their developmental capability. Finally, this study further proves that increasing the freezing and thawing velocity benefits survival of vitrified immature oocytes. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Acupuntura un tratamiento viable para las adicciones en Colombia

    Directory of Open Access Journals (Sweden)

    Hernán López Seuscún

    2013-07-01

    Los tratamientos con auriculoterapia, como el protocolo NADA (National Acupuncture Detoxification Association, son los métodos más usados para las adicciones en el mundo, y aunque no se ha logrado evidenciar su efectividad, por su costo, facilidad y el poco riesgo de efectos adversos se hace viable en un país con pocos recursos económicos como Colombia.

  10. Academic Pediatric Dentistry is a Rewarding, Financially Viable Career Path.

    Science.gov (United States)

    Townsend, Janice A; Chi, Donald L

    2017-09-15

    Newly graduated pediatric dentists have unprecedented levels of debt. High levels of student debt may be perceived as an obstacle to pursue an academic career. However, opportunities exist through faculty compensation models and loan repayment programs that make an academic career financially viable. The purpose of this paper is to outline the benefits of a career in academic dentistry and provide examples of young pediatric dentistry faculty members who have been able to manage student debt while pursuing meaningful and rewarding careers.

  11. How Can We Prevent Violence Becoming a Viable Political Strategy?

    OpenAIRE

    Patricia Justino

    2009-01-01

    A basic issue that conflict analysis investigates is how non-peaceful ways of living and governing become viable political strategies. Macro-level studies provide some important insights but micro-level analysis is vital to understand the mechanisms that make violence possible. This briefing outlines some preliminary findings in this respect from MICROCON, a major research programme analysing violent conflict at the micro level. It also discusses their implications for policies aimed at preve...

  12. Distributions of 14 elements on 60 selected absorbers from two simulant solutions (acid-dissolved sludge and alkaline supernate) for Hanford HLW Tank 102-SY

    Energy Technology Data Exchange (ETDEWEB)

    Marsh, S.F.; Svitra, Z.V.; Bowen, S.M.

    1993-10-01

    Sixty commercially available or experimental absorber materials were evaluated for partitioning high-level radioactive waste. These absorbers included cation and anion exchange resins, inorganic exchangers, composite absorbers, and a series of liquid extractants sorbed on porous support-beads. The distributions of 14 elements onto each absorber were measured from simulated solutions that represent acid-dissolved sludge and alkaline supernate solutions from Hanford high-level waste (HLW) Tank 102-SY. The selected elements, which represent fission products (Ce, Cs, Sr, Tc, and Y); actinides (U, Pu, and Am); and matrix elements (Cr, Co, Fe, Mn, Zn, and Zr), were traced by radionuclides and assayed by gamma spectrometry. Distribution coefficients for each of the 1680 element/absorber/solution combinations were measured for dynamic contact periods of 30 min, 2 h, and 6 h to provide sorption kinetics information for the specified elements from these complex media. More than 5000 measured distribution coefficients are tabulated.

  13. Summary of International Waste Management Programs (LLNL Input to SNL L3 MS: System-Wide Integration and Site Selection Concepts for Future Disposition Options for HLW)

    Energy Technology Data Exchange (ETDEWEB)

    Greenberg, Harris R. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Blink, James A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Halsey, William G. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Sutton, Mark [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2011-08-11

    The Used Fuel Disposition Campaign (UFDC) within the Department of Energy’s Office of Nuclear Energy (DOE-NE) Fuel Cycle Technology (FCT) program has been tasked with investigating the disposal of the nation’s spent nuclear fuel (SNF) and high-level nuclear waste (HLW) for a range of potential waste forms and geologic environments. This Lessons Learned task is part of a multi-laboratory effort, with this LLNL report providing input to a Level 3 SNL milestone (System-Wide Integration and Site Selection Concepts for Future Disposition Options for HLW). The work package number is: FTLL11UF0328; the work package title is: Technical Bases / Lessons Learned; the milestone number is: M41UF032802; and the milestone title is: “LLNL Input to SNL L3 MS: System-Wide Integration and Site Selection Concepts for Future Disposition Options for HLW”. The system-wide integration effort will integrate all aspects of waste management and disposal, integrating the waste generators, interim storage, transportation, and ultimate disposal at a repository site. The review of international experience in these areas is required to support future studies that address all of these components in an integrated manner. Note that this report is a snapshot of nuclear power infrastructure and international waste management programs that is current as of August 2011, with one notable exception. No attempt has been made to discuss the currently evolving world-wide response to the tragic consequences of the earthquake and tsunami that devastated Japan on March 11, 2011, leaving more than 15,000 people dead and more than 8,000 people missing, and severely damaging the Fukushima Daiichi nuclear power complex. Continuing efforts in FY 2012 will update the data, and summarize it in an Excel spreadsheet for easy comparison and assist in the knowledge management of the study cases.

  14. INCONEL 690 CORROSION IN WTP (WASTE TREATMENT PLANT) HLW (HIGH LEVEL WASTE) GLASS MELTS RICH IN ALUMINUM & BISMUTH & CHROMIUM OR ALUMINUM/SODIUM

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; FENG Z; GAN H; PEGG IL

    2009-11-05

    Metal corrosion tests were conducted with four high waste loading non-Fe-limited HLW glass compositions. The results at 1150 C (the WTP nominal melter operating temperature) show corrosion performance for all four glasses that is comparable to that of other typical borosilicate waste glasses, including HLW glass compositions that have been developed for iron-limited WTP streams. Of the four glasses tested, the Bi-limited composition shows the greatest extent of corrosion, which may be related to its higher phosphorus content. Tests at higher suggest that a moderate elevation of the melter operating temperature (up to 1200 C) should not result in any significant increase in Inconel corrosion. However, corrosion rates did increase significantly at yet higher temperatures (1230 C). Very little difference was observed with and without the presence of an electric current density of 6 A/inch{sup 2}, which is the typical upper design limit for Inconel electrodes. The data show a roughly linear relationship between the thickness of the oxide scale on the coupon and the Cr-depletion depth, which is consistent with the chromium depletion providing the material source for scale growth. Analysis of the time dependence of the Cr depletion profiles measured at 1200 C suggests that diffusion of Cr in the Ni-based Inconel alloy controls the depletion depth of Cr inside the alloy. The diffusion coefficient derived from the experimental data agrees within one order of magnitude with the published diffusion coefficient data for Cr in Ni matrices; the difference is likely due to the contribution from faster grain boundary diffusion in the tested Inconel alloy. A simple diffusion model based on these data predicts that Inconel 690 alloy will suffer Cr depletion damage to a depth of about 1 cm over a five year service life at 1200 C in these glasses.

  15. A rapid biosensor for viable B. anthracis spores.

    Science.gov (United States)

    Baeumner, Antje J; Leonard, Barbara; McElwee, John; Montagna, Richard A

    2004-09-01

    A simple membrane-strip-based biosensor assay has been combined with a nucleic acid sequence-based amplification (NASBA) reaction for rapid (4 h) detection of a small number (ten) of viable B. anthracis spores. The biosensor is based on identification of a unique mRNA sequence from one of the anthrax toxin genes, the protective antigen ( pag), encoded on the toxin plasmid, pXO1, and thus provides high specificity toward B. anthracis. Previously, the anthrax toxins activator ( atxA) mRNA had been used in our laboratory for the development of a biosensor for the detection of a single B. anthracis spore within 12 h. Changing the target sequence to the pag mRNA provided the ability to shorten the overall assay time significantly. The vaccine strain of B. anthracis (Sterne strain) was used in all experiments. A 500-microL sample containing as few as ten spores was mixed with 500 microL growth medium and incubated for 30 min for spore germination and mRNA production. Thus, only spores that are viable were detected. Subsequently, RNA was extracted from lysed cells, selectively amplified using NASBA, and rapidly identified by the biosensor. While the biosensor assay requires only 15 min assay time, the overall process takes 4 h for detection of ten viable B. anthracis spores, and is shortened significantly if more spores are present. The biosensor is based on an oligonucleotide sandwich-hybridization assay format. It uses a membrane flow-through system with an immobilized DNA probe that hybridizes with the target sequence. Signal amplification is provided when the target sequence hybridizes to a second DNA probe that has been coupled to liposomes encapsulating the dye sulforhodamine B. The amount of liposomes captured in the detection zone can be read visually or quantified with a hand-held reflectometer. The biosensor can detect as little as 1 fmol target mRNA (1 nmol L(-1)). Specificity analysis revealed no cross-reactivity with 11 organisms tested, among them closely

  16. New device for the vitrification and in-straw warming of in vitro produced bovine embryos.

    Science.gov (United States)

    Morató, Roser; Mogas, Teresa

    2014-04-01

    Two experiments were designed to test the use of a new device designed to vitrify and in-straw warm in vitro produced (IVP) embryos, which can potentially be used for their direct transfer to recipient females in field conditions. In experiment 1, IVP embryos from both prepubertal and adult animals were vitrified on cryotops and warmed in steps (1, 0.5 and 0M sucrose; protocol W3) or directly in 0.5M (protocol W1/0.5) or 0M sucrose (protocol W1/0). Similar survival rates were recorded 24h after warming for calf embryos irrespective of the warming procedure (W3: 79.2%, W1/0.5: 62.5%, W1/0: 66.7%). For cow embryos, survival rates at 24h post-warming were significantly higher when embryos were warmed using the W3 (85.7%) or W1/0.5 (89.1%) protocols compared to the W1/0 protocol (70.5%). In experiment 2, IVP embryos were vitrified on the new designed device followed by their in-straw cryoprotectant (0.5M sucrose) dilution/warming and different warming temperatures (45, 50, 60 and 70°C) were tested. When warming solution passed through the new vitrification/warming device at 45°C, 61.5% of blastocysts were fully re-expanded or hatched at 24h post-warming, being not significantly different to the control (65%). Other warming temperatures triggered significantly lower survival rates at 24h post-warming. No significant differences were detected in total cell numbers and blastocyst apoptosis indices in response to vitrification followed by warming at 45°C respect to the control. Our findings indicate that the new device allows vitrification and in-straw warming of IVP bovine embryos, being a useful option for their direct transfer in field conditions. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Comparison of pregnancy outcomes after vitrification at the cleavage and blastocyst stage: a meta-analysis.

    Science.gov (United States)

    Zeng, MeiFang; Su, SuQin; Li, LiuMing

    2017-09-22

    This systematic review sought to evaluate the clinical outcomes of vitrification at the cleavage stage and blastocyst stage for embryo transfer in patients undergoing assisted reproductive technology (ART) treatment. We searched for related comparative studies published in the PubMed, EMBASE, and Cochrane Library databases up to July 2017. The primary outcomes were clinical pregnancy rate (CPR) and embryo implantation rate (IR). Secondary outcomes were multiple pregnancy rate (MPR), miscarriage rate (MR), live birth rate (LBR), and ongoing pregnancy rate (OPR). The Mantel-Haenszel fixed effects model and random effects model were used to analyze the summary risks ratios (RRs) with 95% confidence intervals (CIs). Eight studies with more than 6590 cycles were included in our meta-analysis. Seven studies were observational retrospective comparative studies. One was a prospective study. Overall, the current study summarizes information from 6590 vitrification warming cycles (cleavage stage n = 4594; blastocysts n = 1996). There was no difference in the primary outcome clinical pregnancy rate (RR = 0.97, 95% CI = 0.90-1.04; fixed effects model; I (2)  = 21%), whereas vitrified blastocyst transfer was significantly superior to vitrified cleavage-stage embryo transfer regarding the implantation rate (RR = 0.85, 95% CI = 0.74-0.97; random effects model; I (2)  = 43). Regarding the secondary outcomes, no differences were found in the multiple pregnancy rate (RR = 1.20, 95% CI = 0.79-1.82; fixed effects model; I (2)  = 22), live birth rate (RR = 1.07, 95% CI = 0.98-1.16; fixed effects model; I (2)  = 0), and ongoing pregnancy rate (RR = 1.01, 95% CI = 0.92-1.120; fixed effects model; I (2)  = 0), whereas a higher miscarriage rate was observed with vitrified blastocyst transfer (RR = 0.65, 95% CI = 0.45-0.93; random effects model; I (2)  = 23). In summary, this meta-analysis shows that vitrification at any stage has no detrimental

  18. Role of cumulus cells during vitrification and fertilization of mature bovine oocytes: Effects on survival, fertilization, and blastocyst development.

    Science.gov (United States)

    Ortiz-Escribano, N; Smits, K; Piepers, S; Van den Abbeel, E; Woelders, H; Van Soom, A

    2016-07-15

    This study was designed to determine the role of cumulus cells during vitrification of bovine oocytes. Mature cumulus-oocyte complexes (COCs) with many layers of cumulus cells, corona radiata oocytes (CRs), with a few layers of cumulus cells, and denuded oocytes (DOs) without cumulus cells were vitrified in 15% ethylene glycol, 15% dimethyl sulfoxide, and 0.5-M sucrose. Oocytes that survived the vitrification process were fertilized. Denuded oocytes were fertilized with or without supplementation of intact COCs (DOsCOCs). First, survival and embryo development rates were studied. Higher survival rates were obtained for DOs and DOsCOCs (94% and 95%, respectively) compared with COCs (82.7%, P vitrification of mature bovine oocytes. Because cumulus cells are required for fertilization, the use of partially DOs (CRs) or the addition of intact COCs (DOsCOCs) during fertilization can result in higher survival and embryo development after vitrification. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Modulation of the endocannabinoid system in viable and non-viable first trimester pregnancies by pregnancy-related hormones

    Directory of Open Access Journals (Sweden)

    Taylor Anthony H

    2011-11-01

    Full Text Available Abstract Background In early pregnancy, increased plasma levels of the endocannabinoid anandamide (AEA are associated with miscarriage through mechanisms that might affect the developing placenta or maternal decidua. Methods In this study, we compare AEA levels in failed and viable pregnancies with the levels of the trophoblastic hormones (beta-human chorionic gonadotrophin (beta-hCG, progesterone (P4 and (pregnancy-associated placental protein-A (PAPP-A essential for early pregnancy success and relate that to the expression of the cannabinoid receptors and enzymes that modulate AEA levels. Results The median plasma AEA level in non-viable pregnancies (1.48 nM; n = 20 was higher than in viable pregnancies (1.21 nM; n = 25; P = 0.013, as were progesterone and beta-hCG levels (41.0 vs 51.5 ng/mL; P = 0.052 for P4 and 28,650 vs 6,560 mIU/L; P = 0.144 for beta-hCG, respectively, but were not statistically significant. Serum PAPP-A levels in the viable group were approximately 6.8 times lower than those in the non-viable group (1.82 vs 12.25 mg/L; P = 0.071, but again these differences were statistically insignificant. In the spontaneous miscarriage group, significant correlations between P4 and beta-hCG, P4 and PAPP-A and AEA and PAPP-A levels were observed. Simultaneously, immunohistochemical distributions of the two main cannabinoid receptors and the AEA-modifying enzymes, fatty acid amide hydrolase (FAAH and N-acylphosphatidylethanolamine-phospholipase D (NAPE-PLD, changed within both the decidua and trophoblast. Conclusions The association of higher AEA levels with early pregnancy failure and with beta-hCG and PAPP-A, but not with progesterone concentrations suggest that plasma AEA levels and pregnancy failure are linked via a mechanism that may involve trophoblastic beta-hCG, and PAPP-A, but not, progesterone production. Although the trophoblast, decidua and embryo contain receptors for AEA, the main AEA target in early pregnancy failure

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

    Science.gov (United States)

    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.

  1. Slow and steady cell shrinkage reduces osmotic stress in bovine and murine oocyte and zygote vitrification.

    Science.gov (United States)

    Lai, D; Ding, J; Smith, G W; Smith, G D; Takayama, S

    2015-01-01

    Does the use of a new cryoprotectant agent (CPA) exchange protocol designed to minimize osmotic stress improve oocyte or zygote vitrification by reducing sublethal cryodamage? The use of a new CPA exchange protocol made possible by automated microfluidics improved oocyte and zygote vitrification with superior morphology as indicated by a smoother cell surface, higher sphericity, higher cytoplasmic lipid retention, less cytoplasmic leakage and higher developmental competence compared with conventional methods. The use of more 'steps' of CPA exposure during the vitrification protocol increases cryosurvival and development in the bovine model. However, such an attempt to eliminate osmotic stress is limited by the practicality of performing numerous precise pipetting steps in a short amount of time. Murine meiotically competent germinal vesicle intact oocytes and zygotes were harvested from the antral follicles in ovaries and ampulla, respectively. Bovine ovaries were obtained from a local abattoir at random stages of the estrous cycle. A total of 110 murine oocytes, 802 murine zygotes and 52 bovine oocytes were used in this study. Microfluidic devices were fabricated using conventional photo- and soft-lithography. CPAs used were 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for equilibration solution and 15% EG, 15% DMSO and 0.5 M sucrose for vitrification solution. End-point analyses include mathematical modeling using Kedem-Katchalsky equations, morphometrics assessed by conventional and confocal microscopy, cytoplasmic lipid quantification by nile red staining, cytoplasmic leakage quantification by fluorescent dextran intercalation and developmental competence analysis by 96 h embryo culture and blastomere quantification. The automated microfluidics protocol decreased the shrinkage rate of the oocyte and zygote by 13.8 times over its manual pipetting alternative. Oocytes and zygotes with a lower shrinkage rate during CPA exposure experienced less

  2. Crystallization and vitrification of electrons in a glass-forming charge liquid

    Science.gov (United States)

    Sasaki, S.; Hashimoto, K.; Kobayashi, R.; Itoh, K.; Iguchi, S.; Nishio, Y.; Ikemoto, Y.; Moriwaki, T.; Yoneyama, N.; Watanabe, M.; Ueda, A.; Mori, H.; Kobayashi, K.; Kumai, R.; Murakami, Y.; Müller, J.; Sasaki, T.

    2017-09-01

    Charge ordering (CO) is a phenomenon in which electrons in solids crystallize into a periodic pattern of charge-rich and charge-poor sites owing to strong electron correlations. This usually results in long-range order. In geometrically frustrated systems, however, a glassy electronic state without long-range CO has been observed. We found that a charge-ordered organic material with an isosceles triangular lattice shows charge dynamics associated with crystallization and vitrification of electrons, which can be understood in the context of an energy landscape arising from the degeneracy of various CO patterns. The dynamics suggest that the same nucleation and growth processes that characterize conventional glass-forming liquids guide the crystallization of electrons. These similarities may provide insight into our understanding of the liquid-glass transition.

  3. Ultrastructure of bovine oocytes exposed to Taxol prior to OPS vitrification

    DEFF Research Database (Denmark)

    Morató, Roser; Mogas, Teresa; Maddox-Hyttel, Poul

    2008-01-01

    for calves: (1) a control group fixed immediately after maturation; (2) an OPS group cryopreserved by conventional OPS; (3) a Taxol/CPA group exposed to 1 microM Taxol and cryoprotective agents (CPAs); and (4) a Taxol/OPS group vitrified by OPS including 1 microM Taxol to the vitrification solution. All...... oocytes were processed for light and transmission electron microscopy. The main injuries were observed on the metaphase plate and the spindle. In control oocytes, the metaphase appeared as condensed chromosomes arranged in a well-organized metaphase plate and the spindle showed well organized microtubules...... in both cow and calf oocytes. However, in cow OPS oocytes, the metaphase plate was disorganized into scattered chromosomes or the chromosomes were condensed into a single block of chromatin. In addition, microtubules were not organized as typical spindles. In contrast, cow Taxol/OPS oocytes as well...

  4. Human Factors engineering criteria and design for the Hanford Waste Vitrification Plant preliminary safety analysis report

    Energy Technology Data Exchange (ETDEWEB)

    Wise, J.A.; Schur, A.; Stitzel, J.C.L.

    1993-09-01

    This report provides a rationale and systematic methodology for bringing Human Factors into the safety design and operations of the Hanford Waste Vitrification Plant (HWVP). Human Factors focuses on how people perform work with tools and machine systems in designed settings. When the design of machine systems and settings take into account the capabilities and limitations of the individuals who use them, human performance can be enhanced while protecting against susceptibility to human error. The inclusion of Human Factors in the safety design of the HWVP is an essential ingredient to safe operation of the facility. The HWVP is a new construction, nonreactor nuclear facility designed to process radioactive wastes held in underground storage tanks into glass logs for permanent disposal. Its design and mission offer new opposites for implementing Human Factors while requiring some means for ensuring that the Human Factors assessments are sound, comprehensive, and appropriately directed.

  5. Test plan for BWID Phase 2 electric arc melter vitrification tests

    Energy Technology Data Exchange (ETDEWEB)

    Soelberg, N.R.; Turner, P.C.; Oden, L.L.; Anderson, G.L.

    1994-10-01

    This test plan describes the Buried Waste Integrated Demonstration (BWID), Phase 2, electric arc melter, waste treatment evaluation tests to be performed at the US Bureau of Mines (USBM) Albany Research Center. The BWID Arc Melter Vitrification Project is being conducted to evaluate and demonstrate existing industrial arc melter technology for thermally treating mixed transuranic-contaminated wastes and soils. Phase 1 baseline tests, performed during fiscal year 1993 at the USBM, were conducted on waste feeds representing incinerated buried mixed wastes and soils. In Phase 2, surrogate feeds will be processed that represent actual as-retrieved buried wastes from the Idaho National Engineering Laboratory`s Subsurface Disposal Area at the Radioactive Waste Management Complex.

  6. Cold-cap reactions in vitrification of nuclear waste glass: experiments and modeling

    Energy Technology Data Exchange (ETDEWEB)

    Chun, Jaehun [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Pierce, David A. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Pokorny, Richard [Inst. of Chemical Technology, Prague (Czech Republic); Hrma, Pavel R. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Pohang Univ. of Science and Techology (Korea, Republic of)

    2013-05-01

    Cold-cap reactions are multiple overlapping reactions that occur in the waste-glass melter during the vitrification process when the melter feed is being converted to molten glass. In this study, we used differential scanning calorimetry (DSC) to investigate cold-cap reactions in a high-alumina high-level waste melter feed. To separate the reaction heat from both sensible heat and experimental instability, we employed the run/rerun method, which enabled us to define the degree of conversion based on the reaction heat and to estimate the heat capacity of the reacting feed. Assuming that the reactions are nearly independent and can be approximated by the nth order kinetics, we obtained the kinetic parameters using the Kissinger method combined with least squares analysis. The resulting mathematical simulation of the cold-cap reactions provides a key element for the development of an advanced cold-cap model.

  7. Vitrification affects nuclear maturation and gene expression of immature human oocytes

    Directory of Open Access Journals (Sweden)

    Abbas Shahedi

    2017-02-01

    Full Text Available Background: Vitrification of oocytes is a fast-freezing technique, which may affect the quality of the human oocyte, and consequently affects the embryo development, pregnancy and birth. The aim of the current study was to investigate the consequence of in-vitro vitrification on maturation status of immature human oocytes, additionally, expression levels of stress, and apoptosis related genes. Materials and Methods: The total of 213 human immature oocytes which routinely discarded from assisted reproduction clinics were collected and divided into two groups including: (I fresh germinal vesicle (GV oocytes (n=106 (matured in-vitro  (fIVM , and  (II GV oocytes (n=107 that initially vitrified, then matured in  in-vitro (vIVM. After 36 hours of incubation, the oocytes were evaluated for nuclear maturation and expression level of DNA methyltransferase (DNMT1, stress related genes (Sod1 and Hsp70, and apoptotic related genes (Bax and Bcl-2 by quantitative Real-Time PCR. Results: Oocyte maturation rates were reduced in vIVM compared to fIVM oocytes (P=0.001. The expression of stress (Sod1 and Hsp70, and apoptotic-related genes (Bax and Bcl-2 in vIVM were significantly higher compared to the fIVM group. Additionally, pro-apoptotic gene up-regulated 4.3 times more than anti-apoptotic gene in vIVM oocyte. However, DNMT1 gene expression was reduced in vIVM oocyte (P = 0.047. Conclusions: The low survival rate of vitrified In-vitro matured GV oocytes could definitely be explained by the alterations of their gene expression profile. 

  8. Improved development by melatonin treatment after vitrification of mouse metaphase II oocytes.

    Science.gov (United States)

    Zhang, Yue; Li, Wei; Ma, Yongshun; Wang, Dian; Zhao, Xiaoxue; Zeng, Changjun; Zhang, Ming; Zeng, Xianyin; Meng, Qinggang; Zhou, Guangbin

    2016-12-01

    The study was aimed to investigate the effect of melatonin on the development potential of mouse MII oocytes after cryopreservation. Mouse MII oocytes were subjected first to vitrification/warming and 2 h of in vitro culture (phase 1), then to parthenogenetic activation (PA) followed by in vitro culture of parthenogenetic embryos (phase 2). Different concentrations of melatonin (0, 10(-9), 10(-6) mol/L) were added to the medium during either phase 1, phase 2 or both phases. The fresh oocytes were used as control. When melatonin was used during both phases, 10(-9) mol/L melatonin-treated group showed similar rates of cleavage and 4-cell embryo development compared with control, which were significantly higher than those of melatonin-free group, while the rates in either 10(-6) mol/L melatonin-treated or melatonin-free groups were significantly lower than that in control. When 10(-9) mol/L melatonin was added during either phase 1 or phase 2, both cleavage and 4-cell embryo development rates of either group were significantly lower than those of control. After oocyte vitrification/warming and PA, the ROS levels increased significantly and maternal-to-zygotic transition (MZT) related genes (Dcp1a, Dcp2, Hspa1a, Eif1ax, Pou5f1, Sox2) expression were disorganized. However, after 10(-9) mol/L melatonin supplementation, the ROS levels decreased significantly compared with melatonin-free group, and the gene expressions were almost recovered to normal level of control group. These results demonstrated that 10(-9) mol/L melatonin supplementation could increase the developmental potential of vitrified-warmed mouse MII oocytes, which may result from ROS scavenging activities and recovery of normal levels of the expressions of MZT-related genes. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Effects of trehalose vitrification and artificial oocyte activation on the development competence of human immature oocytes.

    Science.gov (United States)

    Zhang, Zhiguo; Wang, Tianjuan; Hao, Yan; Panhwar, Fazil; Chen, Zhongrong; Zou, Weiwei; Ji, Dongmei; Chen, Beili; Zhou, Ping; Zhao, Gang; Cao, Yunxia

    2017-02-01

    Sucrose and trehalose are conventional cryoprotectant additives for oocytes and embryos. Ethanol can artificially enhance activation of inseminated mature oocytes. This study aims to investigate whether artificial oocyte activation (AOA) with ethanol can promote the development competence of in vitro matured oocytes. A total of 810 human immature oocytes, obtained from 325 patients undergoing normal stimulated oocyte retrieval cycles, were in vitro maturated (IVM) either immediately after collection (Fresh group n = 291)) or after being vitrified as immature oocytes (Vitrified group n = 519). These groups were arbitrarily assigned. All fresh and vitrified oocytes which matured after a period of IVM then underwent intra-cytoplasmic sperm injection (ICSI). Half an hour following ICSI, they were either activated by 7% ethanol (AOA group) or left untreated (Non-AOA group). Fertilization, cleavage rate, blastocyst quality and aneuploidy rate were then evaluated. High-quality blastocysts were only obtained in both the fresh and vitrified groups which had undergone AOA after ICSI. Trehalose vitrification slightly, but not significantly, increased the formation rates of high-quality embryos (21.7% VS 15.4%, P > 0.05) and blastocysts (15.7% VS 7.69%, P > 0.05)) when compared with sucrose vitrification. Aneuploidy was observed in 12 of 24 (50%) of the AOA derived high quality blastocysts. High-quality blastocysts only developed from fresh or vitrified immature oocytes if the ICSI was followed by AOA. This information may be important for human immature oocytes commonly retrieved in normal stimulation cycles and may be particularly important for certain patient groups, such as cancer patients. AOA with an appropriate concentration of ethanol can enhance the developmental competence of embryos. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Inkjet printing of viable human dental follicle stem cells

    Directory of Open Access Journals (Sweden)

    Mau Robert

    2015-09-01

    Full Text Available Inkjet printing technology has the potential to be used for seeding of viable cells for tissue engineering approaches. For this reason, a piezoelectrically actuated, drop-on-demand inkjet printing system was applied to deliver viable human dental follicle stem cells (hDFSC of sizes of about 15 μm up to 20 μm in diameter. The purpose of these investigations was to verify the stability of the printing process and to evaluate cell viability post printing. Using a Nanoplotter 2.1 (Gesim, Germany equipped with the piezoelectric printhead NanoTip HV (Gesim, Germany, a concentration of 6.6 ×106 cells ml−1 in DMEM with 10% fetal calf serum (FCS could be dispensed. The piezoelectric printhead has a nominal droplet volume of ~ 400 pl and was set to a voltage of 75 V and a pulse of 50 μs while dosing 50 000 droplets over a time of 100 seconds. The volume and trajectory of the droplet were checked by a stroboscope test right before and after the printing process. It was found that the droplet volume decreases significantly by 35% during printing process, while the trajectory of the droplets remains stable with only an insignificant number of degrees deviation from the vertical line. It is highly probable that some cell sedimentations or agglomerations affect the printing performance. The cell viability post printing was assessed by using the Trypan Blue dye exclusion test. The printing process was found to have no significant influence on cell survival. In conclusion, drop-on-demand inkjet printing can be a potent tool for the seeding of viable cells.

  11. Effect of vitrification on the zona pellucida hardening and follistatin and cathepsin B genes expression and developmental competence of in vitro matured bovine oocytes.

    Science.gov (United States)

    Wiesak, Teresa; Wasielak, Marta; Złotkowska, Aleksandra; Milewski, Robert

    2017-06-01

    The purpose of our study was to assess the effect of vitrification with or without the presence of calcium in the vitrification solution on the: 1) diameter of oocytes and thickness of the zona pellucida, 2) zona pellucida hardening, 3) expression of mRNA follistatin (FST) and cathepsin B (CTSB) in oocytes and 4) developmental competence of embryos derived from in vitro matured and vitrified oocytes. The results of our study demonstrate, that vitrification did not alter thickness of the zona pellucida and diameter of the oocytes, however it triggered hardening of the zona pellucida. The presence of calcium in the vitrification solutions intensified hardening of zona in immature and mature oocytes (P vitrification did not change the oocyte diameter and thickness of the zona pellucida and expression of FST and CTSB mRNA. It diminished developmental potential of the vitrified oocytes. The presence of calcium in the vitrification solutions increased hardening of zona pellucida as well as affected the level of FST and CTSB mRNA in oocytes and developmental potential of these oocytes. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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.

  13. Cellular alteration after dilution of cryoprotective solutions used for the vitrification of in vitro-produced bovine embryos.

    Science.gov (United States)

    Kaidi, S; Van Langendonckt, A; Massip, A; Dessy, F; Donnay, I

    1999-08-01

    Embryo quality of in vitro-produced bovine blastocysts was assessed at several steps of a vitrification procedure in which glycerol and ethylene glycol were used as cryoprotectants (3-step equilibration with cryoprotectants followed by vitrification, dilution of the cryoprotectants in 0.85 M galactose then in embryo transfer freezing medium [ETF], and finally co-culture for periods). To visualize cell membrane alterations, double staining was performed using a cell permeant fluorochrome (bisbenzimide--BIS) and a nonpermeant one (propidium iodide--PI). In Experiment 1, the effect of the vitrification procedure on the hatching rate and total cell number was assessed 72 h after treatment. Hatching rate and the number of stained nuclei were decreased in comparison with untreated embryos when blastocysts were exposed to the whole procedure with or without vitrification (respectively 42 and 53% vs 76% for hatching and 128 +/- 17 and 141 +/- 17 vs 226 +/- 13 for stained nuclei). In Experiment 2, the effect of cryoprotectants and their dilution was evaluated on membrane permeability and total cell numbers at various steps of the vitrification procedure. Blastocysts exposed only to cryoprotectant solutions and stained immediately after dilution in galactose showed no modification. After dilution in ETF, the total number of stained nuclei decreased, and the number of blastomeres showing membrane permeabilization (PI-stained) increased (respectively, 74 +/- 5 vs 110 +/- 5 and 32 +/- 2% vs 0.1 +/- 1.8%). In Experiment 3, we demonstrated that the total number of stained nuclei after ethanol fixation (membrane permeabilization) was higher when embryos treated up to dilution in ETF were stained with PI than when the same embryos were stained with BIS. This suggests that, for unknown reasons, some nuclei of the treated embryos were not stained with BIS. Membrane permeabilization and inability of BIS to stain some nuclei were the most obvious alterations probably induced by osmotic

  14. Molar Pregnancy with a Co-Existing Viable Fetus

    Directory of Open Access Journals (Sweden)

    Ruya Deveer

    2014-03-01

    Full Text Available     The aim of this study was to report the clinical features, management, and outcome of a case of molar pregnancy with a coexisting viable fetus and to review the literature. In this article, we report a case of pregnancy with diffuse placental molar change and a normal fetus which presented with hyperemesis gravidarum and hyperthyroidism. Genetic amniocentesis showed normal fetal karyotype. A healthy full-term live male infant was delivered by cesarean section. In molar pregnancies with a normal karyotype fetus, with intensive maternal follow-up, continuation of pregnancy can be suggested.

  15. Radioactive waste combustion / vitrification under arc plasma: thermal and dynamic modelling; Combustion - vitrification de dechets radioactifs par plasma d'arc: modelisation de la thermique et de la dynamique

    Energy Technology Data Exchange (ETDEWEB)

    Barthelemy, B

    2003-07-01

    This thesis concerns the thermal and dynamic modelling for a combustion/vitrification process of surrogate radioactive waste under transferred arc plasma. The writer presents the confinement processes for radioactive waste using arc plasma and the different software used to model theses processes. This is followed by a description of our experimental equipment including a plasma arc reactor and an inductive system allowing the homogenization of glass temperature. A combustion/vitrification test is described. Thermal and material balances were discussed. The temperature fields of plasma arc and the glass frit conductivity are measured. Finally, the writer describes and clarifies the equations solved for the simulations of the electrically plasma arc and the glass melting including the thin layer of glass frit coating the crucible cold walls. The modelling results are presented in the form of spatial distribution of temperature, velocity and volume power... (author)

  16. Radioactive waste combustion-vitrification under arc plasma: thermal and dynamic modelling; Combustion - vitrification de dechets radioactifs par plasma d'arc: modelisation de la thermique et de la dynamique

    Energy Technology Data Exchange (ETDEWEB)

    Barthelemy, B

    2003-06-01

    This thesis concerns the thermal and dynamic modelling for a combustion/vitrification process of surrogate radioactive waste under transferred arc plasma. The writer presents the confinement processes for radioactive waste using arc plasma and the different software used to model theses processes. This is followed by a description of our experimental equipment including a plasma arc reactor and an inductive system allowing the homogenization of glass temperature. A combustion/vitrification test is described. Thermal and material balances were discussed. The temperature fields of plasma arc and the glass frit conductivity are measured. Finally, the writer describes and clarifies the equations solved for the simulations of the electrically plasma arc and the glass melting including the thin layer of glass frit coating the crucible cold walls. The modelling results are presented in the form of spatial distribution of temperature, velocity and voluminal power... (author)

  17. The effect of vitrification of immature bovine oocytes to the subsequent in vitro development and gene expression.

    Science.gov (United States)

    Faheem, Marwa S; Baron, E; Carvalhais, I; Chaveiro, A; Pavani, K; da Silva, F Moreira

    2015-12-01

    Immature bovine oocytes were vitrified using the cryotop method and their post-warming survivability and capability to undergo in vitro maturation, fertilization and subsequent embryonic development were evaluated. In addition throughout the embryonic 2-cell, 4-cell, morula and blastocyst stages, the expression of four developmentally important genes (Cx43, CDH1, DNMT1 and HSPA14) was analysed using the real-time polymerase chain reaction (PCR). Immature oocytes (n = 550) were randomly assigned to non-vitrified (fresh) or cryotop vitrification groups using ethylene glycol (EG) with 1,2 propanediol (PROH) or dimethylsulphoxide (DMSO). After warming, oocytes survivability, embryo cleavage and embryonic developmental rates were not statistically different between the two cryoprotectants groups. However, the DMSO group had a lower (P vitrification of immature bovine oocytes, both for embryonic developmental competence and at the molecular level. Moreover, PROH showed some advantage over DMSO as a cryoprotectant.

  18. Evaluation of melter technologies for vitrification of Hanford site low-level tank waste - phase 1 testing summary report

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, C.N., Westinghouse Hanford

    1996-06-27

    Following negotiation of the fourth amendment to the Tri- Party Agreement for Hanford Site cleanup, commercially available melter technologies were tested during 1994 and 1995 for vitrification of the low-level waste (LLW) stream to be derived from retrieval and pretreatment of the radioactive defense wastes stored in 177 underground tanks. Seven vendors were selected for Phase 1 testing to demonstrate vitrification of a high-sodium content liquid LLW simulant. The tested melter technologies included four Joule-heated melters, a carbon electrode melter, a combustion melter, and a plasma melter. Various dry and slurry melter feed preparation processes also were tested. The technologies and Phase 1 testing results were evaluated and a preliminary technology down-selection completed. This report describes the Phase 1 LLW melter vendor testing and the tested technologies, and summarizes the testing results and the preliminary technology recommendations.

  19. High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification

    DEFF Research Database (Denmark)

    Du, Y; Pribenszky, C S; Molnár, M

    2008-01-01

    The purpose of the present study was to improve cryotolerance using high hydrostatic pressure (HHP) pretreatment of porcine in vitro matured (IVM) oocytes, to facilitate their further developmental competence after parthenogenetic activation. A total of 1668 porcine IVM oocytes were used in our...... present study. The pressure tolerance and optimal duration of recovery after HHP treatment were determined. Oocytes were treated with either 20 or 40 MPa (200 and 400 times greater than atmospheric pressure) for 60 min, with an interval of 10, 70, and 130 min between pressure treatment and subsequent...... vitrification under each pressure parameter. Oocytes from all vitrification groups had much lower developmental competence than fresh oocytes (Ppressure, with either 70...

  20. Cryopreservation of in vitro shoot tips of strawberry by the vitrification method - establishment of a duplicate collection of Fragaria germplasm.

    Science.gov (United States)

    Höfer, Monika

    2016-01-01

    The National German Strawberry Genebank, which includes 183 cultivars and 270 accessions of wild Fragaria species, is maintained in a field collection of the Fruit Genebank at Dresden-Pillnitz. A duplicate collection stored in liquid nitrogen would provide a higher level of security for these irreplaceable genetic resources. Four distinct cryopreservation protocols were tested earlier and the best method, a PVS2 vitrification method with a 14-day alternating-temperature cold acclimation, was modified for further genotype screening. A comprehensive genotype spectrum was tested by screening 107 Fragaria ×ananassa cultivars and 20 Fragaria wild species (51 accessions). The average recovery for the cultivars using the optimized medium was 89.55 % and for the wild species 85.50 %. These results indicate that the method could be used to establish a backup collection in liquid nitrogen. Meristem cultures will be established and the collection will be cryopreserved using this vitrification technique to provide cost effective long-term storage.

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

    Directory of Open Access Journals (Sweden)

    Safrina Rahmah

    2015-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

    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; Melter 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

  3. Embryo vitrification using a novel semi-automated closed system yields in vitro outcomes equivalent to the manual Cryotop method.

    Science.gov (United States)

    Roy, Tammie K; Brandi, Susanna; Tappe, Naomi M; Bradley, Cara K; Vom, Eduardo; Henderson, Chester; Lewis, Craig; Battista, Kristy; Hobbs, Ben; Hobbs, Simon; Syer, John; Lanyon, Sam R; Dopheide, Sacha M; Peura, Teija T; McArthur, Steven J; Bowman, Mark C; Stojanov, Tomas

    2014-11-01

    Can the equilibration steps prior to embryo vitrification be automated? We have developed the 'Gavi' system which automatically performs equilibration steps before closed system vitrification on up to four embryos at a time and gives in vitro outcomes equivalent to the manual Cryotop method. Embryo cryopreservation is an essential component of a successful assisted reproduction clinic, with vitrification providing excellent embryo survival and pregnancy outcomes. However, vitrification is a manual, labour-intensive and highly skilled procedure, and results can vary between embryologists and clinics. A closed system whereby the embryo does not come in direct contact with liquid nitrogen is preferred by many clinics and is a regulatory requirement in some countries. The Gavi system, an automation instrument with a novel closed system device, was used to equilibrate embryos prior to vitrification. Outcomes for embryos automatically processed with the Gavi system were compared with those processed with the manual Cryotop method and with fresh (non-vitrified) controls. The efficacy of the Gavi system (Alpha model) was assessed for mouse (Quackenbush Swiss and F1 C57BL/6J x CBA) zygotes, cleavage stage embryos and blastocysts, and for donated human vitrified-warmed blastocysts. The main outcomes assessed included recovery, survival and in vitro embryo development after vitrification-warming. Cooling and warming rates were measured using a thermocouple probe. Mouse embryos vitrified after processing with the automated Gavi system achieved equivalent in vitro outcomes to that of Cryotop controls. For example, for mouse blastocysts both the Gavi system (n = 176) and manual Cryotop method (n = 172) gave a 99% recovery rate, of which 54 and 50%, respectively, progressed to fully hatched blastocysts 48 h after warming. The outcomes for human blastocysts processed with the Gavi system (n = 23) were also equivalent to Cryotop controls (n = 13) including 100% recovery for both

  4. Ultra-Structural Alterations in In Vitro Produced Four-Cell Bovine Embryos Following Controlled Slow Freezing or Vitrification.

    Science.gov (United States)

    Cavusoglu, T; Popken, J; Guengoer, T; Yilmaz, O; Uyanikgil, Y; Ates, U; Baka, M; Oztas, E; Zakhartchenko, V

    2016-08-01

    Cryopreservation is the process of freezing and preserving cells and tissues at low temperatures. Controlled slow freezing and vitrification have successfully been used for cryopreservation of mammalian embryos. We investigated the effect of these two cryopreservation methods on in vitro produced four-cell stage bovine embryos which were classified according to their quality and separated into three groups. The first group was maintained as untreated controls (n = 350). Embryos of the second (n = 385) and the third (n = 385) groups were cryopreserved either by controlled slow freezing or by vitrification. Embryos in groups 2 and 3 were thawed after 1 day. Hundred embryos were randomly selected from the control group, and 100 morphologically intact embryos from the second and third group were thawed after 1 day and cultured to observe the development up to the blastocyst stage. The blastocyst development rate was 22% in the control group, 1% in the slow-freezing group and 3% in the vitrification group. Remaining embryos of all three groups were examined by light microscopy, transmission electron microscopy and immunofluorescence confocal microscopy with subsequent histological staining procedures. Cryopreservation caused degenerative changes at the ultra-structural level. Compared with vitrification, slow freezing caused an increased mitochondrial degeneration, cytoplasmic vacuolization, disruption of the nuclear and plasma membrane integrity, organelle disintegration, cytoskeletal damage, a reduced thickness of the zona pellucida and a formation of fractures in the zona pellucida. Further studies are required to understand and decrease the harmful effects of cryopreservation. © 2015 Blackwell Verlag GmbH.

  5. Coenzyme Q10 supplementation during in vitro maturation of bovine oocytes (Bos taurus) helps to preserve oocyte integrity after vitrification.

    Science.gov (United States)

    Ruiz-Conca, M; Vendrell, M; Sabés-Alsina, M; Mogas, T; Lopez-Bejar, M

    2017-10-01

    Oocyte vitrification causes less cell stress than slow cooling, but cytoskeletal and spindle alterations may occur affecting the oocyte competence. In vitro maturation (IVM) supplementation with different antioxidant molecules has been performed to attenuate this harmful stress. Coenzyme Q10 (CoQ10 ) supplementation has previously shown to have positive effects in bovine and mouse in vitro embryo development. The aim of this study was to evaluate the effects of CoQ10 during bovine oocyte IVM and vitrification. Cumulus-oocyte complexes (COCs) (n = 311) were cultured under standard maturation conditions with 0 μM (control), 25 μM and 50 μM CoQ10 supplementation. After 22 hr, a cohort of 170 oocytes both from the control and from CoQ10 -supplemented groups were vitrified, warmed and returned to incubation until 24 hr of maturation, while the rest of the oocytes (n = 141) remained fresh. Then, oocyte survival was assessed morphologically by stereomicroscopy. Oocytes from all groups were then fixed and stained for assessing cortical granules (CG) migration and nuclear stage. High rates of oocyte MII progression and appropriate CG migration as a continuous layer beneath the plasma membrane were obtained both in control and in CoQ10 groups. Results showed that although vitrification has great impact in survival of IVM bovine oocytes, 50 μM CoQ10 supplementation significantly improved oocyte survival (p = .045) and reduced the premature CG exocytosis, helping to preserve the CG migration pattern (31.3% control vs. 54.5% in 50 μM CoQ10 ; p = .039), attenuating the negative effects of vitrification. © 2017 Blackwell Verlag GmbH.

  6. Vitrification of in vitro-produced bovine embryos by addition of ethylene glycol in one-step.

    Science.gov (United States)

    Walker, D J; Campos-Chillon, L F; Seidel, G E

    2006-10-01

    The objective of this study was to simplify two-step addition of cryoprotectant for vitrification of bovine embryos by developing a one-step procedure. Survival was calculated as a percentage of non-vitrified controls developed from the same batch of oocytes. In experiment 1, bovine blastocysts were vitrified following one- or two-step addition of cryoprotectant. Exposure of embryos to cryoprotectant in one-step resulted in survival rates not significantly lower (p > 0.1) than those obtained by two-step addition (85% vs 98%, respectively). Based on these results, experiments 2-4 were designed to test one-step addition of cryoprotectant more rigorously. Experiment 2 exposed day 7 blastocysts to 6, 7 or 8 M ethylene glycol for 2.5 or 3.5 min. At 24 h post-vitrification, survival of embryos was similar, irrespective of ethylene glycol concentration or exposure time (6 M 38%, 7 M 51%, 8 M 59%; 2.5 min 54%, 3.5 min 45%). In experiment 3, blastocysts were exposed to 7 M ethylene glycol for shorter times (30 or 60 s); 30 s exposure resulted in decreased survival (8% vs 31%, p bovine morulae, exposed to 7 M ethylene glycol for 1 or 1.5 min. There was no difference in survival between exposure times of 1 or 1.5 min (28% vs 45%, respectively; p > 0.1). It is unclear why many embryos survive vitrification with one-step addition of cryoprotectant, but others do not. Although, one-step addition of cryoprotectant simplifies the vitrification procedure, survival rates were inadequate for routine cryopreservation of in vitro-produced bovine embryos.

  7. Comparison on in vitro fertilized bovine embryos cultured in KSOM or SOF and cryopreserved by slow freezing or vitrification.

    Science.gov (United States)

    Nedambale, T L; Dinnyés, A; Groen, W; Dobrinsky, J R; Tian, X C; Yang, X

    2004-08-01

    The objectives of this study were to identify an improved in vitro cell-free embryo culture system and to compare post-warming development of in vitro produced (IVP) bovine embryos following vitrification versus slow freezing. In Experiment 1, non-selected presumptive zygotes were randomly allocated to four medium treatments without co-culture: (1) SOF + 5% FCS for 9 days; (2) KSOM + 0.1% BSA for 4 days and then KSOM + 1% BSA to Day 9; (3) SOF + 5% FCS for 4 days and then KSOM + 1% BSA to Day 9; and (4) KSOM + 0.1% BSA for 4 days and then SOF + 5% FCS to Day 9. Treatment 4 (sequential KSOM-SOF culture system) improved (P > 0.05) morulae (47%), early blastocysts (26%), Day-7 blastocysts (36%), cell numbers, as well as total hatching rate (79%) compared to KSOM alone (Treatment 2). Embryos cultured in KSOM + BSA alone developed slowly and most of them hatched late on Day 9, compared to other treatments. In Experiment 2, the sequential KSOM-SOF culture system was used and Day-7 blastocysts were subjected to following cryopreservation comparison: (1) vitrification (VS3a, 6.5 M glycerol); or (2) slow freezing (1.36 M glycerol). Warmed embryos were cultured in SOF with 7.5% FCS. Higher embryo development and hatching rates (P vitrification at 6h (71%), 24h (64%), and 48h (60%) post-warming compared to slow freezing (48, 40, and 31%, respectively). Following transfer of vitrified embryos to synchronized recipients, a 30% pregnancy rate was obtained. In conclusion, replacing KSOM with SOF after 4 days of culture produced better quality blastocysts. Vitrification using VS3a may be used more effectively to cryopreserve in vitro produced embryos than the conventional slow freezing method.

  8. Improved cryopreservation of oil palm (Elaeis guineensis Jacq.) polyembryoids using droplet vitrification approach and assessment of genetic fidelity.

    Science.gov (United States)

    Gantait, Saikat; Sinniah, Uma Rani; Suranthran, Periasamy; Palanyandy, Sharrmila Rengeswari; Subramaniam, Sreeramanan

    2015-01-01

    In the present study, polyembryoids of oil palm (Elaeis guineensis Jacq.) were cryopreserved with successful revival of 68 % for the first time using the droplet vitrification technique. Excised polyembryoids (3-5-mm diameter) from 3-month-old in vitro cultures were pre-cultured for 12 h in liquid Murashige and Skoog medium supplemented with 0.5 M sucrose. The polyembryoids were osmoprotected in loading solution [10% (w/v) dimethyl sulphoxide (DMSO) plus 0.7 M sucrose] for 30 min at room temperature and then placed on aluminium strips where they were individually drenched in chilled droplets of vitrification solution (PVS2) [30% (w/v) glycerol plus 15% (w/v) ethylene glycol (EG) plus 15% (w/v) DMSO plus 0.4 M sucrose] for 10 min. The aluminium strips were enclosed in cryovials which were then plunged quickly into liquid nitrogen and kept there for 1 h. The polyembryoids were then thawed and unloaded (using 1.2 M sucrose solution) with subsequent transfer to regeneration medium and stored in zero irradiance. Following for 10 days of storage, polyembryoids were cultured under 16 h photoperiod of 50 μmol m(-2) s(-1) photosynthetic photon flux density, at 23 ± 1 °C. Post-thaw growth recovery of 68% was recorded within 2 weeks of culture, and new shoot development was observed at 4 weeks of growth. Scanning electron microscopy revealed that successful regeneration of cryopreserved polyembryoids was related to maintenance of cellular integrity, presumably through PVS2 exposure for 10 min. The present study demonstrated that cryopreservation by droplet vitrification enhanced the regeneration percentages of oil palm in comparison with the conventional vitrification method previously reported.

  9. Vitrification of in vitro matured oocytes collected from antral follicles at the time of ovarian tissue cryopreservation

    Directory of Open Access Journals (Sweden)

    Fasano Giovanna

    2011-11-01

    Full Text Available Abstract Background In the past few years, cryopreservation of ovarian tissue has become an established procedure proposed in many centers around the world and transplantation has successfully resulted in full-term pregnancies and deliveries in human. This prospective study aims to evaluate the feasibility of vitrifying in vitro matured oocytes (IVM isolated at the time of ovarian tissue cryopreservation to improve the efficiency of fertility preservation programs. Methods Oocyte-cumulus complexes were retrieved from freshly collected ovarian cortex by aspirating antral follicular fluid, and were matured in vitro for 24-48 h prior to vitrification. Oocytes were matured in an IVM commercial medium (Copper Surgical, USA supplemented with 75 mIU/ml FSH and 75 mIU/ml LH and vitrified using a commercial vitrification kit (Irvine Scientific, California in high security vitrification straws (CryoBioSystem, France. Oocyte collection and IVM rates were evaluated according to the age, the cycle period and the amount of tissue collected. Results Immature oocyte retrieval from ovarian tissue was carried out in 57 patients between 8 and 35 years of age, undergoing ovarian tissue cryopreservation. A total of 266 oocytes were isolated, 28 of them were degenerated, 200 were at germinal vesicle stage (GV, 35 were in metaphase I (MI and 3 displayed a visible polar body (MII. The number of oocytes collected was positively correlated with the amount of tissue cryopreserved (p p = 0.005. Oocytes were obtained regardless of menstrual cycle period or contraception. A total maturation rate of 31% was achieved, leading to the vitrification of at least one mature oocyte for half of the cohort. Conclusions The study showed that a significant number of immature oocytes can be collected from excised ovarian tissue whatever the menstrual cycle phases and the age of the patients, even for prepubertal girls.

  10. The Roles of Vitrification of Stabilizers/Matrix Formers for the Redispersibility of Drug Nanocrystals After Solidification: a Case Study.

    Science.gov (United States)

    Yue, PengFei; Xiao, MingSheng; Xie, YuanBiao; Ma, YueQin; Guan, YongMei; Wu, ZhenFeng; Hu, PengYi; Wang, YaQi

    2016-12-01

    To elucidate the roles of vitrification of stabilizers/matrix formers for the redispersibility of drug nanocrystal powder after solidification at storage stress, the influence of different drying methods and storage stresses on stability of drug nanocrystals was systemically investigated. A poorly soluble drug, baicalin, used as model drug was converted into baicalin nanocrystals (BCN-NC). The residual moisture contents of BCN-NC were applied at two different stress conditions defined as "conservative" (1%), respectively. The influence of different stabilizers, matrix formers, and storage stresses on the redispersibility of BCN-NC powder was systemically investigated, respectively. The results showed that storage stresses had significantly influence the redispersibility of BCN-NC. Aggressive storage temperature and residual moisture could be unfavorable factors for stability of drug nanocrystals, due to the exacerbation of aggregation of BCN-NC induced by vitrification. It was demonstrated that vitrification of spray-dried BCN-NC was dependent on temperature and time. The polymeric stabilizers hydroxypropylmethylcellulose (HPMC) and sodium carboxymethyl starch (CMS-Na) with high glass transition temperature (T g) played more important role in protecting the BCN-NC from breakage during storage, compared to the surfactants Tween 80, D-α-tocopherol acid polyethylene glycol 1000 succinate (TPGS), or RH 40. Besides, the polyvinylpyrrolidone K30 (PVP K30) and lactose with high T g were effective matrix formers for preserving the redispersibility of BCN-NC. It was concluded that the vitrification transition of stabilizers/matrix formers could be responsible for aggregation of drug nanocrystals during storage, which was a time-dependent process. The suitable residual moisture contents (RMC) and T g were very important for preserving the stability of drug nanocrystals during storage.

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

    Directory of Open Access Journals (Sweden)

    Wen-yan Song

    2016-05-01

    Full Text Available Background/Aims: 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. Methods: 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. Results: 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 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 P P > 0.05. Rate of comet cells was significantly lower in group A than in groups B and C (all P P Conclusion: 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.

  12. FINAL REPORT INTEGRATED DM1200 MELTER TESTING OF BUBBLER CONFIGURATIONS USING HLW AZ-101 SIMULANTS VSL-04R4800-4 REV 0 10/5/04

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; GONG W; BARDAKCI T; D' ANGELO NA; LUTZE W; CALLOW RA; BRANDYS M; KOT WK; PEGG IL

    2011-12-29

    This report documents melter and off-gas performance results obtained on the DM1200 HLW Pilot Melter during processing of AZ-101 HLW simulants. The tests reported herein are a subset of six tests from a larger series of tests described in the Test Plan for the work; results from the other tests have been reported separately. The solids contents of the melter feeds were based on the WTP baseline value for the solids content of the feeds from pretreatment which changed during these tests from 20% to 15% undissolved solids resulting in tests conducted at two feed solids contents. Based on the results of earlier tests with single outlet 'J' bubblers, initial tests were performed with a total bubbling rate of 651 pm. The first set of tests (Tests 1A-1E) addressed the effects of skewing this total air flow rate back and forth between the two installed bubblers in comparison to a fixed equal division of flow between them. The second set of tests (2A-2D) addressed the effects of bubbler depth. Subsequently, as the location, type and number of bubbling outlets were varied, the optimum bubbling rate for each was determined. A third (3A-3C) and fourth (8A-8C) set of tests evaluated the effects of alternative bubbler designs with two gas outlets per bubbler instead of one by placing four bubblers in positions simulating multiple-outlet bubblers. Data from the simulated multiple outlet bubblers were used to design bubblers with two outlets for an additional set of tests (9A-9C). Test 9 was also used to determine the effect of small sugar additions to the feed on ruthenium volatility. Another set of tests (10A-10D) evaluated the effects on production rate of spiking the feed with chloride and sulfate. Variables held constant to the extent possible included melt temperature, plenum temperature, cold cap coverage, the waste simulant composition, and the target glass composition. The feed rate was increased to the point that a constant, essentially complete, cold cap was

  13. Effects of antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification on the in vitro developmental capacity of expanded bovine blastocysts.

    Science.gov (United States)

    Liang, Shuang; Yuan, Bao; Jin, Yong-Xun; Zhang, Jia-Bao; Bang, Jeong Kyu; Kim, Nam-Hyung

    2017-10-01

    Cryopreservation is an effective method for the long-term storage of valuable germplasm in the field of reproductive research. The present study examined the developmental capacity of post-thaw bovine blastocysts during vitrification after supplementation with antifreeze glycoprotein 8 (AFGP8). Survival and re-expansion rates in culture during the 12h after thawing were significantly higher in the AFGP8-treated than untreated group. In addition, blastocysts from the AFGP8-treated group exhibited lower rates of apoptosis. Real-time reverse transcription-polymerase chain reaction analysis showed that the expression of the Bcl-2 gene, coding for an anti-apoptotic protein, was increased significantly, whereas the expression of the pro-apoptotic gene Bax was decreased significantly in the AFGP8-treated group. The cellular proliferation rate and mitochondrial membrane potential were significantly higher in post-thaw re-expanded blastocysts from the AFGP8-treated compared with untreated group. In addition, outgrowth potential in post-thaw blastocysts in re-expanded blastocysts after vitrification was significantly increased in the AFGP8-treated compared with untreated group. Together, these results are the first to demonstrate that the addition of AFGP8 during vitrification can help protect bovine blastocysts against chill-induced injury.

  14. Vitrification of in vitro produced bovine embryos at different ages using one- and three-step addition of cryoprotective additives.

    Science.gov (United States)

    Saha, S; Suzuki, T

    1997-01-01

    The effect of embryo age on development and ratio of live: dead cells after vitrification and warming was examined. One-step and three-step addition of cryoprotectants in vitrification solution (40% ethylene glycol, 0.3 M trehalose and 12% polyvinylpyrrolidone) were compared using in vitro produced (IVP) bovine blastocysts and expanded blastocysts. Rates of development and hatching were 74.2% and 41.9% for Day 7, 57.8% and 23.8% for Day 8, 33.7% and 6.1% for Day 9 embryos with one-step addition. In three-step addition, those rates were 86.2% and 77.3% for Day 7, 72.3% and 39.0% for Day 8, 47.3% and 10.5% for Day 9 embryos. Day 7 embryos showed highest (P embryos with three-step addition was higher (P embryos (P bovine embryos against vitrification and the potential for three-step addition of cryoprotectants to yield a higher survival rate after warming than with one-step addition.

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

    Science.gov (United States)

    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 (Pvitamins 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.

  16. Social Networking and Smart Technology: Viable Environmental Communication Tools…?

    Science.gov (United States)

    Montain, J.; Byrne, J. M.

    2010-12-01

    To what extent do popular social networking channels represent a viable means for disseminating information regarding environmental change to the general public? Are new forms of communication such as YouTube™, Facebook™, MySpace™ and Twitter™ and smart devices such as iPhone™ and BlackBerry™ useful and effective in terms motivating people into social action and behavioural modification; or do they simply pay ‘lip service’ to these pressing environmental issues? This project will explore the background connections between social networking and environmental communication and education; and outline why such tools might be an appropriate way to connect to a broad audience in an efficient and unconventional manner. Further, research will survey the current prevalence of reliable environmental change information on social networking Internet-based media; and finally, suggestions for improved strategies and new directions will be provided.

  17. Dissolvable tattoo sensors: from science fiction to a viable technology

    Science.gov (United States)

    Cheng, Huanyu; Yi, Ning

    2017-01-01

    Early surrealistic painting and science fiction movies have envisioned dissolvable tattoo electronic devices. In this paper, we will review the recent advances that transform that vision into a viable technology, with extended capabilities even beyond the early vision. Specifically, we focus on the discussion of a stretchable design for tattoo sensors and degradable materials for dissolvable sensors, in the form of inorganic devices with a performance comparable to modern electronics. Integration of these two technologies as well as the future developments of bio-integrated devices is also discussed. Many of the appealing ideas behind developments of these devices are drawn from nature and especially biological systems. Thus, bio-inspiration is believed to continue playing a key role in future devices for bio-integration and beyond.

  18. A viable logarithmic f(R) model for inflation

    Energy Technology Data Exchange (ETDEWEB)

    Amin, M.; Khalil, S. [Center for Fundamental Physics, Zewail City of Science and Technology,6 October City, Giza (Egypt); Salah, M. [Center for Fundamental Physics, Zewail City of Science and Technology,6 October City, Giza (Egypt); Department of Mathematics, Faculty of Science, Cairo University,Giza (Egypt)

    2016-08-18

    Inflation in the framework of f(R) modified gravity is revisited. We study the conditions that f(R) should satisfy in order to lead to a viable inflationary model in the original form and in the Einstein frame. Based on these criteria we propose a new logarithmic model as a potential candidate for f(R) theories aiming to describe inflation consistent with observations from Planck satellite (2015). The model predicts scalar spectral index 0.9615

  19. Simultaneous pyometra and viable puppies’ gestation in a bitch

    Directory of Open Access Journals (Sweden)

    A. Risso

    2014-08-01

    Full Text Available Here we describe a case of pyometra coexisting with gestation in a 4.5 year-old miniature short-haired Dachshund. The dog exhibited depression, vaginal discharge, polydipsia and dehydration. Ultrasound examination revealed the presence of low to moderate anechoic fluid collection in the left uterine horn. Blood analysis revealed mild neutrophilia with a left shift. Based on these findings a presumptive diagnosis of pyometra was made and the bitch was treated using amoxicillin-clavulanate with dopaminergic agonist (cabergoline. A second ultrasound scan revealed the presence of two gestational vesicles in the right uterine horn that were successfully carried to term. Unusually, while pyometra persisted in the left uterine horn, two viable puppies were delivered by caesarean section from the right uterine horn.

  20. Profiling Total Viable Bacteria in a Hemodialysis Water Treatment System.

    Science.gov (United States)

    Chen, Lihua; Zhu, Xuan; Zhang, Menglu; Wang, Yuxin; Lv, Tianyu; Zhang, Shenghua; Yu, Xin

    2017-05-28

    Culture-dependent methods, such as heterotrophic plate counting (HPC), are usually applied to evaluate the bacteriological quality of hemodialysis water. However, these methods cannot detect the uncultured or viable but non-culturable (VBNC) bacteria, both of which may be quantitatively predominant throughout the hemodialysis water treatment system. Therefore, propidium monoazide (PMA)-qPCR associated with HPC was used together to profile the distribution of the total viable bacteria in such a system. Moreover, high-throughput sequencing of 16S rRNA gene amplicons was utilized to analyze the microbial community structure and diversity. The HPC results indicated that the total bacterial counts conformed to the standards, yet the bacteria amounts were abruptly enhanced after carbon filter treatment. Nevertheless, the bacterial counts detected by PMA-qPCR, with the highest levels of 2.14 × 10 7 copies/100 ml in softener water, were much higher than the corresponding HPC results, which demonstrated the occurrence of numerous uncultured or VBNC bacteria among the entire system before reverse osmosis (RO). In addition, the microbial community structure was very different and the diversity was enhanced after the carbon filter. Although the diversity was minimized after RO treatment, pathogens such as Escherichia could still be detected in the RO effluent. In general, both the amounts of bacteria and the complexity of microbial community in the hemodialysis water treatment system revealed by molecular approaches were much higher than by traditional method. These results suggested the higher health risk potential for hemodialysis patients from the up-to-standard water. The treatment process could also be optimized, based on the results of this study.

  1. Vitrification and Product Testing of C-104 and AZ-102 Pretreated Sludge Mixed with Flowsheet Quantities of Secondary Wastes

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Gary L.; Bates, Derrick J.; Goles, Ronald W.; Greenwood, Lawrence R.; Lettau, Ralph C.; Piepel, Gregory F.; Schweiger, Michael J.; Smith, Harry D.; Urie, Michael W.; Wagner, Jerome J.

    2001-02-01

    The U.S. Department of Energy (DOE) Office of River Protection (ORP) has acquired Hanford tank waste treatment services at a demonstration scale. The River Protection Project Waste Treatment Plant (RPP-WTP) team is responsible for producing an immobilized (vitrified) high-level waste (IHLW) waste form. Pacific Northwest National Laboratory, hereafter referred to as PNNL, has been contracted to produce and test a vitrified IHLW waste form from two Envelope D high-level waste (HLW) samples previously supplied to the RPP-WTP project by DOE.

  2. Factors affecting survival rates of in vitro produced bovine embryos after vitrification and direct in-straw rehydration.

    Science.gov (United States)

    Vajta, G; Holm, P; Greve, T; Callesen, H

    1996-12-16

    The aim of this work was to investigate the possibilities of simplification, and to outline the limits of application, of a vitrification method for cow embryos. Morulae and blastocysts were produced by in vitro fertilization of slaughterhouse-derived, in vitro matured oocytes with frozen-thawed bull semen, and subsequent culture on a granulosa cell monolayer. Vitrification was performed by equilibration of embryos with 12.5% ethylene glycol and 12.5% dimethylsulphoxide at 20-22 degrees C for 60 s, then with 25% ethylene glycol and 25% dimethylsulphoxide at 4 degrees C for another 60 s. Embryos were then loaded in straws, placed in liquid nitrogen vapour for 2 min, and then plunged. Straws were thawed in a 22 degrees C water-bath, the embryos were directly rehydrated and further incubated in straw, and were then expelled and cultured in vitro for 72 h. In the first experiment, embryos of different age and developmental stage (Day 5 compacted morulae, Day 6 early blastocysts, Days 6 and 7 blastocysts, Day 7 expanded blastocysts and Day 8 hatched blastocysts) as well as Days 7 and 5 blastocysts previously subjected to partial zone dissection were vitrified. After thawing, the re-expansion rates of blastocysts and zona-dissected embryos did not differ (67 and 87%, respectively), and hatching was more frequent for blastocysts frozen in advanced developmental stages (34, 47 and 63% for early blastocysts, blastocysts and expanded blastocysts, respectively). The re-expansion rate of morulae was lower (10%) and no hatching of these embryos was observed. In the second experiment, Day 7 expanded blastocysts were vitrified using PBS, PBS+albumin, TCM199 and TCM199+calf serum as holding media. No differences in re-expansion and hatching rates were seen. However, when incubation with the concentrated cryoprotectant solution was performed at 20-22 degrees C, the embryo survival rate decreased (PBS+albumin) or no embryo survived (TCM199+calf serum) the vitrification procedure. In

  3. The effect of media, serum and temperature on in vitro survival of bovine blastocysts after Open Pulled Straw (OPS) vitrification.

    Science.gov (United States)

    Vajta, G; Rindom, N; Peura, T T; Holm, P; Greve, T; Callesen, H

    1999-10-01

    The recently introduced Open Pulled Straw (OPS) vitrification technique has successfully been used for cryopreserving porcine embryos as well as for bovine embryos and oocytes. The aim of this work is to investigate several factors on the in vitro survival of bovine blastocysts. In 5 experiments, a total of 862 in vitro produced blastocysts and expanded blastocysts was vitrified and warmed using the OPS technology, then cultured in vitro for an additional 3 days. The culture medium in Experiments 1 to 4 was SOFaa with supplements and 5% calf serum (CS). In Experiment 1, the replacement of TCM-199 + 20% CS with PBS + 20% CS in the holding medium during vitrification and warming did not result in significant differences in the re-expansion (92 vs 95%) and hatching rates (79 vs 72%). In Experiment 2, the PBS holding medium was supplemented with either 20% CS, 5 mg/mL bovine serum albumin (BSA) or 3 mg/mL polyvinylalcohol (PVA). Although the re-expansion rates did not differ (98, 95 and 93%, respectively), there was a decrease in the hatching rate after vitrification with PVA (77 and 78 vs 51%, respectively). In Experiment 3, the influence of temperature of equilibration media prior to and rehydration media after the vitrification was investigated. When the temperature of these media was adjusted to 20 degrees C instead of the standard 35 degrees C, both the re-expansion and the hatching rates decreased markedly. However, increasing the time of equilibration with the diluted cryoprotectant solution at 20 degrees C eliminated these differences. In Experiment 4, the ethylene-glycol and dimethyl sulfoxide cryoprotectant mixture was replaced with ethylene glycol-ficoll-trehalose solution. No difference in the re-expansion (89 vs 96%, respectively) or hatching rate (79 vs 84%, respectively) was detected. In Experiment 5, the vitrified-warmed blastocysts were cultured in SOFaa medium supplemented with 5% CS or 5 mg/mL BSA. Although the re-expansion rates were identical in

  4. Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification

    Directory of Open Access Journals (Sweden)

    Melissa Meneghel

    Full Text Available ABSTRACT: The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification. Embryos were produced from abattoir-derived ovaries and allocated into four groups. In the treatment groups, forskolin was added to the in vitro culture medium on Day 6 and incubated for 24 hours in one of the following concentrations: 2.5μM (Forsk 2.5 group, 5.0μM (Forsk 5.0 group or 10.0μM (Forsk 10.0 group. Embryos from the control group were cultured without forskolin. On Day 7 of culture, the expanded blastocysts were stained with the lipophilic dye Sudan Black B for determination of the intracytoplasmic lipid content or were cryopreserved via the Vitri-Ingá® procedure. Although there were no significant differences (P>0.05 in the blastocyst rates between the Control group (44.9% and the other treatments, the embryo production was lower (P0.05 to that found in Forsk 2.5 (0.92±0.03 and Forsk 10.0 groups (1.06±0.03 groups; however the lipid accumulation in blastocysts from Forsk 5.0 group (0.82±0.04 was lower than in the Control group (P<0.05. Based on these results, Forsk 5.0 treatment was tested for cryotolerance and it was observed that the blastocoel re-expansion rate evaluated 24 hours after warming was greater (P<0.05 in Forsk 5.0 group (72.2% compared to the Control group (46.2%. In conclusion, pre-treatment with forskolin at a concentration of 5.0 μM for 24 hours before vitrification is effective in reducing the intracytoplasmic lipid content and, consequently, improves cryotolerance of IVP bovine embryos.

  5. FINAL REPORT INTEGRATED DM1200 MELTER TESTING OF REDOX EFFECTS USING HLW AZ-101 AND C-106/AY-102 SIMULANTS VSL-04R4800-1 REV 0 5/6/

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; GONG W; BARDAKCI T; D' ANGELO NA; LUTZE W; BIZOT PM; CALLOW RA; BRANDYS M; KOT WK; PEGG IL

    2011-12-29

    This report documents melter and off-gas performance results obtained on the DM1200 HLW Pilot Melter during processing of AZ-101 and C-106/AY-102 HLW simulants. The tests reported herein are a subset of three tests from a larger series of tests described in the Test Plan for the work; results from the remaining tests will be reported separately. Three nine day tests, one with AZ-101 and two with C-106/AY-102 feeds were conducted with variable amounts of added sugar to address the effects of redox. The test with AZ-101 included ruthenium spikes to also address the effects of redox on ruthenium volatility. One of tests addressed the effects of increased flow-sheet nitrate levels using C-106/AY-102 feeds. With high nitrate/nitrite feeds (such as WTP LAW feeds), reductants are required to prevent melt foaming and deleterious effects on glass production rates. Sugar is the baseline WTP reductant for this purpose. WTP HLW feeds typically have relatively low nitrate/nitrite content in comparison to the organic carbon content and, therefore, have typically not required sugar additions. However, HLW feed variability, particularly with respect to nitrate levels, may necessitate the use of sugar in some instances. The tests reported here investigate the effects of variable sugar additions to the melter feed as well as elevated nitrate levels in the waste. Variables held constant to the extent possible included melt temperature, bubbling rate, plenum temperature, cold cap coverage, the waste simulant composition, and the target glass composition. The principal objectives of the DM1200 melter testing were to determine the achievable glass production rates for simulated HLW feeds with variable amounts of added sugar and increased nitrate levels; characterize melter off-gas emissions; characterize the performance of the prototypical off-gas system components as well as their integrated performance; characterize the feed, glass product, and off-gas effluents; and perform pre- and

  6. Addition of L-ascorbic acid to culture and vitrification media of IVF porcine blastocysts improves survival and reduces HSPA1A levels of vitrified embryos.

    Science.gov (United States)

    Castillo-Martín, Miriam; Yeste, Marc; Soler, Albert; Morató, Roser; Bonet, Sergi

    2015-09-01

    The aim of the present study was to determine the effect of L-ascorbic acid on embryo quality and gene expression of porcine blastocysts after supplementations of in vitro culture medium and/or vitrification-warming media. Embryo quality, in terms of total cell number (TCN), DNA fragmentation and peroxide levels, together with the relative transcript abundance of BCL-2 associated X protein (BAX), BCL2-like 1 (BCL2L1), POU class 5 homeobox 1 (POU5F1) and heat shock protein 70 (HSPA1A), was analysed. In Experiment 1, gene expression and embryo quality of fresh blastocysts were evaluated after culture with or without L-ascorbic acid; no significant differences were observed between the groups. In Experiment 2, blastocysts cultured with or without L-ascorbic acid were vitrified using two different vitrification solutions, supplemented or not with L-ascorbic acid. Supplementation of culture and vitrification media significantly enhanced survival rates and reduced peroxide levels. No significant differences in TCN, DNA fragmentation and BAX, BCL2L1 and POU5F1 expression were found in vitrified blastocysts among experimental groups. Vitrification procedures increase HSPA1A transcript abundance, but this increase was significantly lower in embryos cultured and/or vitrified with L-ascorbic acid. Thus, supplementing culture and/or vitrification media with L-ascorbic acid enhances survival rates of porcine blastocysts, suggesting a relationship with HSPA1A expression.

  7. Innovative fossil fuel fired vitrification technology for soil remediation. Phase 1

    Energy Technology Data Exchange (ETDEWEB)

    1994-01-01

    Vortec has successfully completed Phase 1 of the ``Innovative Fossil Fuel Fired Vitrification Technology for Soil Remediation`` program. The Combustion and Melting System (CMS) has processed 7000 pounds of material representative of contaminated soil that is found at DOE sites. The soil was spiked with Resource Conservation and Recovery Act (RCRA) metals surrogates, an organic contaminant, and a surrogate radionuclide. The samples taken during the tests confirmed that virtually all of the radionuclide was retained in the glass and that it did not leach to the environment-as confirmed by both ANS 16.1 and Toxicity Characteristic Leaching Procedure (TCLP) testing. The organic contaminant, anthracene, was destroyed during the test with a Destruction and Removal Efficiency (DRE) of at least 99.99%. RCRA metal surrogates, that were in the vitrified product, were retained and did not leach to the environment as confirmed by the TCLP testing. Semi-volatile RCRA metal surrogates were captured by the Air Pollution Control (APC) system, and data on the amount of metal oxide particulate and the chemical composition of the particulate were established for use in the Phase 2 APC subsystem design.

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

    Science.gov (United States)

    Park, Min Jee; Lee, Seung Eun; Kim, Eun Young; Lee, Jun Beom; Jeong, Chang Jin; Park, Se Pill

    2015-06-01

    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 (pvitrification groups [enucleated-vitrified-thawed (EVT) group, 13.7%; VT group, 15.0%; pvitrification groups (pvitrification groups than in the non-V group (pvitrification 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.

  9. Investigation of Tc Migration Mechanism During Bulk Vitrification Process Using Re Surrogate

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Dong-Sang; Bagaasen, Larry M.; Crum, Jarrod V.; Fluegel, Alex; Gallegos, Autumn B.; Martinez, Baudelio; Matyas, Josef; Meyer, Perry A.; Paulsen, Dan; Riley, Brian J.; Schweiger, Michael J.; Stewart, Charles W.; Swoboda, Robert G.; Yeager, John D.

    2006-12-04

    As a part of Bulk vitrification (BV) performance enhancement tasks, Laboratory scoping tests were performed in FY 2004-2005 to explore possible ways to reduce the amount of soluble Tc in the BV waste package. Theses scoping tests helped identify which mechanisms play an important role in the migration of Tc in the BV process (Hrma et al. 2005 and Kim et al. 2005). Based on the results from these scoping tests, additional tests were identified that will improve the understanding of Tc migration and to clearly identify the dominant mechanisms. The additional activities identified from previous studies were evaluated and prioritized for planning for Tasks 29 and 30 conducted in FY2006. Task 29 focused on the improved understanding of Tc migration mechanisms, and Task 30 focused on identifying the potential process changes that might reduce Tc/Re migration into the castable refractory block (CRB). This report summarizes the results from the laboratory- and crucible-scale tests in the lab for improved Tc migration mechanism understanding utilizing Re as a surrogate performed in Task 29.

  10. Technical issues associated with in situ vitrification of the INEL Subsurface Disposal Area

    Energy Technology Data Exchange (ETDEWEB)

    Stoots, C.M.; Bates, S.O.; Callow, R.A.; Campbell, K.A.; Farnsworth, R.K.; Gratson, G.K.; McKellar, M.G.; Nickelson, D.F.; Slater, C.E.

    1992-01-01

    In situ vitrification (ISV) has been identified as an alternative technology for remediation of the Acid Pit and Transuranic Pits and Trenches (TRU-PTs) that are present at the Idaho National Engineering Laboratory (INEL) Subsurface Disposal Area (SDA). However, a number of technical issues exist that must be resolved before ISV can be considered applicable to these waste sites. To assist in the ISV technology evaluation, an ISV Steering Committee was formed to identify, prioritize, and develop closure roadmaps for technical issues associated with ISV application at the INEL SDA. The activities of the ISV Steering Committee are summarized in three volumes of this report. Volume 1 identifies the systematic approach used to identify and prioritize the ISV technical issues, and briefly discusses the methodology that will be employed to resolve these issues. This document Volume 2 and Volume 3 discusses each technical issue in greater detail and suggest specific closure roadmaps to be used in resolving technical issues associated with ISV at the SDA Acid Pit and TRU-PTs, respectively.

  11. Vitrification: an alternative to minimize environmental impact caused by leather industry wastes.

    Science.gov (United States)

    Basegio, T; Beck Leão, A P; Bernardes, A M; Bergmann, C P

    2009-06-15

    The main purpose of this work was the investigation of the immobilization of chromium ion present in the ash from the incineration of chromium-tanned leather shavings (ACS) by means of vitrified ceramic bodies. To achieve the immobilization samples were initially produced adding soda-lime glass to ACS. After that, new formulations were produced with the addition of pure oxides, TiO(2) and MgO, to the soda-lime glass and ACS sample. These samples were conformed by pressing and fired in an eletrical furnace, at temperatures of 750, 800, 950 and 1000 degrees C. The ceramic bodies were evaluated with respect to their physical properties and mineralogical composition by X-ray diffraction and energy dispersive spectroscopy (EDS) mapping. The chromium immobilization was characterized by leaching tests according to Brazilian Regulations (NBR 10005). The results confirmed the immobilization of the chromium ion within the allowed limits of Brazilian Regulations, NBR 10004 (5mg/L), with the use of glass and vitrification/densification agents such as titanium and magnesium oxides.

  12. Carbon nanomaterials enhance survival of Agapanthus praecox callus after cryopreservation by vitrification.

    Science.gov (United States)

    Chen, S M; Ren, L; Zhang, D; Zhang, Y F; Shen, X H

    Adding exogenous compounds is an effective way to improve cell survival after cryopreservation. Carbon nanomaterials (CNMs) are novel exogenous substances with small particle size and good biocompatibility. In this work, four types of CNMs were used for the cryopreservation of Agapanthus praecox callus and their possible effects and mechanism of action were analyzed. The thermal properties of the vitrification solutions tested were detected by differential scanning calorimetry. Raman spectroscopy and transmission electron microscopy analyses were used to study the distribution of CNMs inside cells. The MDA/H2O2 contents were measured to evaluate the toxicity of CNMs to cells. Supplementation of PVS2 with various CNMs at different concentrations could enhance survival. The most effective concentration was 0.3 g/L C60, which increased survival by 159% compared to untreated controls and decreased the MDA and H2O2 contents. Single-walled carbon nanotubes (SWCNTs) and C60 entered callus cells. C60 was found only in mitochondria, whereas SWCNTs were located only around the cell walls.

  13. Glass optimization for vitrification of Hanford Site low-level tank waste

    Energy Technology Data Exchange (ETDEWEB)

    Feng, X.; Hrma, P.R.; Westsik, J.H. Jr. [and others

    1996-03-01

    The radioactive defense wastes stored in 177 underground single-shell tanks (SST) and double-shell tanks (DST) at the Hanford Site will be separated into low-level and high-level fractions. One technology activity underway at PNNL is the development of glass formulations for the immobilization of the low-level tank wastes. A glass formulation strategy has been developed that describes development approaches to optimize glass compositions prior to the projected LLW vitrification facility start-up in 2005. Implementation of this strategy requires testing of glass formulations spanning a number of waste loadings, compositions, and additives over the range of expected waste compositions. The resulting glasses will then be characterized and compared to processing and performance specifications yet to be developed. This report documents the glass formulation work conducted at PNL in fiscal years 1994 and 1995 including glass formulation optimization, minor component impacts evaluation, Phase 1 and Phase 2 melter vendor glass development, liquidus temperature and crystallization kinetics determination. This report also summarizes relevant work at PNNL on high-iron glasses for Hanford tank wastes conducted through the Mixed Waste Integrated Program and work at Savannah River Technology Center to optimize glass formulations using a Plackett-Burnam experimental design.

  14. Vitrification of surrogate mixed wastes in a graphite electrode arc melter

    Energy Technology Data Exchange (ETDEWEB)

    Soelberg, N.R.; Chambers, A.G.; Ball, L. [and others

    1995-11-01

    Demonstration tests for vitrifying mixed wastes and contaminated soils have been conducted using a small (800 kVA), industrial-scale, three-phase AC, graphite electrode furnace located at the Albany Research Center of the United States Bureau of Mines (USBM). The feed mixtures were non-radioactive surrogates of various types of mixed (radioactive and hazardous), transuranic-contaminated wastes stored and buried at the Idaho National Engineering Laboratory (INEL). The feed mixtures were processed with added soil from the INEL. Objectives being evaluated include (1) equipment capability to achieve desired process conditions and vitrification products for different feed compositions, (2) slag and metals tapping capability, (3) partitioning of transuranic elements and toxic metals among the furnace products, (4) slag, fume, and metal products characteristics, and (5) performance of the feed, furnace and air pollution control systems. The tests were successfully completed in mid-April 1995. A very comprehensive process monitoring, sampling and analysis program was included in the test program. Sample analysis, data reduction, and results evaluation are currently underway. Initial results indicate that the furnace readily processed around 20,000 lb of widely ranging feed mixtures at feedrates of up to 1,100 lb/hr. Continuous feeding and slag tapping was achieved. Molten metal was also tapped twice during the test program. Offgas emissions were efficiently controlled as expected by a modified air pollution control system.

  15. Iron Mössbauer redox and relation to technetium retention during vitrification

    Science.gov (United States)

    Muller, Isabelle S.; Viragh, Carol; Gan, Hao; Matlack, Keith S.; Pegg, Ian L.

    2009-06-01

    Technetium-99 is a long half-life radioactive fission product whose high volatility during waste vitrification (National Research Council, 2001; Migge, Mat Res Symp Proc 127:205-213, 1989) is a factor in the safe disposal of radioactive waste. Oxidation states of +7, +4 mostly, and some traces of metal have been observed in the glass (Migge, Mat Res Symp Proc 176:411-417, 1990; Darab and Smith, Chem Mater 8:1004-1021, 1996) with Tc7 + more volatile than Tc4 + , leading to a range of technetium retention in the glass varying from 0.8% to 92%. In the present experiments, a simulated high-sulfate Hanford tank waste was vitrified using a slurry fed joule heated furnace producing about 30 kg of glass per day. Various combinations of sucrose and urea were used as denitration and reducing agents in order to optimize sulfate and technetium retention. Since iron oxide is used as a glass additive, Fe-Mössbauer spectroscopy was a vital tool to monitor the melt redox through its Fe2 + /Fetot ratio. A Mössbauer glass redox scale was designed to relate the technetium speciation to its retention in the glass.

  16. Computer modeling of fluid flow and combustion in the ISV (In Situ Vitrification) confinement hood

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, R.W.; Paik, S.

    1990-09-01

    Safety and suitability objectives for the application of the In Situ Vitrification (ISV) technology at the INEL require that the physical processes involved in ISVV be modeled to determine their operational behavior. The mathematical models that have been determined to address the modeling needs adequately for the ISV analysis package are detailed elsewhere. The present report is concerned with the models required for simulating the reacting flow that occurs in the ISV confinement hood. An experimental code named COYOTE has been secured that appears adequate to model the combustion in the confinement hood. The COYOTE code is a two-dimensional, transient, compressible, Eulerian, gas dynamics code for modeling reactive flows. It recognizes nonuniform Cartesian and cylindrical geometry and is based on the ICE (Implicit Continuous-fluid Eulerian) family of solution methods. It includes models for chemical reactions based on chemical kinetics as well as equilibrium chemistry. The mathematical models contained in COYOTE, their discrete analogs, the solution procedure, code structure and some test problems are presented in the report. 12 refs., 17 figs., 6 tabs.

  17. Evaluation of defense-waste glass produced by full-scale vitrification equipment

    Energy Technology Data Exchange (ETDEWEB)

    Lukacs, J.M.; Petkus, L.L.; Mellinger, G.B.

    1981-09-01

    Three full-scale vitrification processes at the Pacific Northwest Laboratory produced over 67,000 kg of simulated nuclear-waste glass from March 1979 to August 1980. Samples were analyzed to monitor process operation and evaluate the resulting glass product. These processes are: Spray Calciner/In-Can Melter (SC/ICM); Spray Calciner/Calcine-Fed Ceramic Melter (SC/CFCM); and Liquid-Fed Ceramic Melter (LFCM). Waste components in the process feed varied less than +- 10%. The SC/ICM and SC/CFCM which use separate waste and frit feed systems showed larger glass compositional variation than the LFCM, which processed only premixed feed during this period. The SC/ICM and SC/CFCM product contained significant amounts of acmite crystals, while the LFCM product was largely amorphous. In addition, the lower portion of all SC/ICM-filled canisters contained a zone rich in waste components. A product chemical durability as determined by pH4 and soxhlet leach tests varied considerably. Aside from increased durability under pH4 conditions with decreasing waste content, glass composition, microstructure and melting process did not correlate with glass durability. For all samples analyzed, the weight loss under pH4 conditions ranged from 17.7 to 85.2 wt %. Soxhlet conditions produced weight losses from 1.78 to 3.56 wt %.

  18. The elusive minimum viable population size for white sturgeon

    Energy Technology Data Exchange (ETDEWEB)

    Jager, Yetta [ORNL; Lepla, Ken B. [Idaho Power Company; Van Winkle, Webb [Van Windle Environmental Consulting; James, Mr Brad [Washington Department of Fish and Wildlife; McAdam, Dr Steve [University of British Columbia, Vancouver

    2010-01-01

    Biological conservation of sturgeon populations is a concern for many species. Those responsible for managing the white sturgeon (Acipenser transmontanus) and similar species are interested in identifying extinction thresholds to avoid. Two thresholds that exist in theory are the minimum viable population size (MVP) and minimum amount of suitable habitat. In this paper, we present both model and empirical estimates of these thresholds. We modified a population viability analysis (PVA) model for white sturgeon to include two new Allee mechanisms. Despite this, PVA-based MVP estimates were unrealistically low compared with empirical estimates unless opportunities for spawning were assumed to be less frequent. PVA results revealed a trade-off between MVP and habitat thresholds; smaller populations persisted in longer river segments and vice versa. Our empirical analyses suggested (1) a MVP range based on population trends from 1,194 to 27,700 individuals, and (2) a MVP estimate of 4,000 individuals based on recruitment. Long-term historical population surveys are needed for more populations to pinpoint an MVP based on trends, whereas the available data were sufficient to estimate MVP based on recruitment. Beyond the MVP, we developed a hierarchical model for population status based on empirical data. Metapopulation support was the most important predictor of population health, followed by the length of free-flowing habitat, with habitat thresholds at 26 and 150 km. Together, these results suggest that habitat and connectivity are important determinants of population status that likely influence the site-specific MVP thresholds.

  19. Towards a viable and just global nursing ethics.

    Science.gov (United States)

    Crigger, Nancy J

    2008-01-01

    Globalization, an outgrowth of technology, while informing us about people throughout the world, also raises our awareness of the extreme economic and social disparities that exist among nations. As part of a global discipline, nurses are vitally interested in reducing and eliminating disparities so that better health is achieved for all people. Recent literature in nursing encourages our discipline to engage more actively with social justice issues. Justice in health care is a major commitment of nursing; thus questions in the larger sphere of globalization, justice and ethics, are our discipline's questions also. Global justice, or fairness, is not an issue for some groups or institutions, but a deeper human rights issue that is a responsibility for everyone. What can we do to help reduce or eliminate the social and economic disparities that are so evident? What kind of ethical milieu is needed to address the threat that globalization imposes on justice and fairness? This article enriches the conceptualization of globalization by investigating recent work by Schweiker and Twiss. In addition, I discuss five qualities or characteristics that will facilitate the development of a viable and just global ethic. A global ethic guides all people in their response to human rights and poverty. Technology and business, two major forces in globalization that are generally considered beneficial, are critiqued as barriers to social justice and the common good.

  20. Keeping checkpoint/restart viable for exascale systems.

    Energy Technology Data Exchange (ETDEWEB)

    Riesen, Rolf E.; Bridges, Patrick G. (IBM Research, Ireland, Mulhuddart, Dublin); Stearley, Jon R.; Laros, James H., III; Oldfield, Ron A.; Arnold, Dorian (University of New Mexico, Albuquerque, NM); Pedretti, Kevin Thomas Tauke; Ferreira, Kurt Brian; Brightwell, Ronald Brian

    2011-09-01

    Next-generation exascale systems, those capable of performing a quintillion (10{sup 18}) operations per second, are expected to be delivered in the next 8-10 years. These systems, which will be 1,000 times faster than current systems, will be of unprecedented scale. As these systems continue to grow in size, faults will become increasingly common, even over the course of small calculations. Therefore, issues such as fault tolerance and reliability will limit application scalability. Current techniques to ensure progress across faults like checkpoint/restart, the dominant fault tolerance mechanism for the last 25 years, are increasingly problematic at the scales of future systems due to their excessive overheads. In this work, we evaluate a number of techniques to decrease the overhead of checkpoint/restart and keep this method viable for future exascale systems. More specifically, this work evaluates state-machine replication to dramatically increase the checkpoint interval (the time between successive checkpoint) and hash-based, probabilistic incremental checkpointing using graphics processing units to decrease the checkpoint commit time (the time to save one checkpoint). Using a combination of empirical analysis, modeling, and simulation, we study the costs and benefits of these approaches on a wide range of parameters. These results, which cover of number of high-performance computing capability workloads, different failure distributions, hardware mean time to failures, and I/O bandwidths, show the potential benefits of these techniques for meeting the reliability demands of future exascale platforms.

  1. Is Greenberg’s “Macro-Carib” viable?

    Directory of Open Access Journals (Sweden)

    Spike Gildea

    2007-08-01

    Full Text Available In his landmark work Language in the Americas, Greenberg (1987 proposed that Macro-Carib was one of the major low-level stocks of South America, which together with Macro-Panoan and Macro-Ge-Bororo were claimed to comprise the putative Ge-Pano-Carib Phylum. His Macro-Carib includes the isolates Andoke and Kukura, and the Witotoan, Peba-Yaguan, and Cariban families. Greenberg’s primary evidence came from person-marking paradigms in individual languages, plus scattered words from individual languages collected into 79 Macro-Carib ‘etymologies’ and another 64 Amerind ‘etymologies’. The goal of this paper is to re-evaluate Greenberg’s Macro-Carib claim in the light of the much more extensive and reliable language data that has become available largely since 1987. Based on full person-marking paradigms for Proto-Cariban, Yagua, Bora and Andoke, we conclude that Greenberg’s morphological claims are unfounded. For our lexical comparison, we created lexical lists for Proto-Cariban, Proto-Witotoan, Yagua and Andoke, for both Greenberg’s 143 putative etymologies and for the Swadesh 100 list. From both lists, a total of 23 potential cognates were found, but no consonantal correspondences were repeated even once. We conclude that our greatly expanded and improved database does not provide sufficient evidence to convince the skeptic that the Macro-Carib hypothesis is viable.

  2. Case-based anatomy teaching: a viable alternative?

    Science.gov (United States)

    Eseonu, Onyedikachi; Carachi, Robert; Brindley, Nicola

    2013-08-01

    Over the last two decades, there has been a decline in the amount of time available for anatomy teaching in the medical undergraduate curriculum, and new methods of anatomy teaching have been adopted for pragmatic reasons, with little evidence base to support their proposed educational benefits. This study seeks to establish the effect of a case-based teaching method on students' confidence in anatomy. Forty-three student volunteers in the clinical phase of the Glasgow medical course were given weekly anatomy teaching sessions based on clinical case presentations over 4 weeks. The students were given an anatomy test, and were asked to rate their confidence in their anatomy knowledge before and after the teaching sessions. There was a two-point increase in students' self-rated confidence, and a 10.9 per cent increase in average test score after the case-based anatomy teaching sessions. Both of these increases were statistically significant (p teaching was also highly rated by students, which may make it a viable option for the teaching of anatomy in the modern medical curriculum. © 2013 John Wiley & Sons Ltd.

  3. Protein design algorithms predict viable resistance to an experimental antifolate.

    Science.gov (United States)

    Reeve, Stephanie M; Gainza, Pablo; Frey, Kathleen M; Georgiev, Ivelin; Donald, Bruce R; Anderson, Amy C

    2015-01-20

    Methods to accurately predict potential drug target mutations in response to early-stage leads could drive the design of more resilient first generation drug candidates. In this study, a structure-based protein design algorithm (K* in the OSPREY suite) was used to prospectively identify single-nucleotide polymorphisms that confer resistance to an experimental inhibitor effective against dihydrofolate reductase (DHFR) from Staphylococcus aureus. Four of the top-ranked mutations in DHFR were found to be catalytically competent and resistant to the inhibitor. Selection of resistant bacteria in vitro reveals that two of the predicted mutations arise in the background of a compensatory mutation. Using enzyme kinetics, microbiology, and crystal structures of the complexes, we determined the fitness of the mutant enzymes and strains, the structural basis of resistance, and the compensatory relationship of the mutations. To our knowledge, this work illustrates the first application of protein design algorithms to prospectively predict viable resistance mutations that arise in bacteria under antibiotic pressure.

  4. Towards viable cosmological models of disformal theories of gravity

    Science.gov (United States)

    Sakstein, Jeremy

    2015-01-01

    The late-time cosmological dynamics of disformal gravity are investigated using dynamical systems methods. It is shown that in the general case there are no stable attractors that screen fifth forces locally and simultaneously describe a dark energy dominated universe. Viable scenarios have late-time properties that are independent of the disformal parameters and are identical to the equivalent conformal quintessence model. Our analysis reveals that configurations where the Jordan frame metric becomes singular are only reached in the infinite future, thus explaining the natural pathology resistance observed numerically by several previous works. The viability of models where this can happen is discussed in terms of both the cosmological dynamics and local phenomena. We identify a special parameter tuning such that there is a new fixed point that can match the presently observed dark energy density and equation of state. This model is unviable when the scalar couples to the visible sector but may provide a good candidate model for theories where only dark matter is disformally coupled.

  5. SMA actuators: a viable practical technology (Presentation Video)

    Science.gov (United States)

    Browne, Alan L.; Brown, Jeffrey; Hodgson, Darel E.

    2015-04-01

    Diverse products either based solely on or incorporating Shape Memory Alloys (SMA) have and are being made in a wide range of industries, and IP is being captured. Why then compared to SE (superelastic) Nitinol, and especially conventional technology, do so few ideas reach production? This presentation delves deeply into this topic in reaching the final assessment that SMA actuators are indeed now a viable practical technology. The presentation begins with an introduction to and description of the fundamental basis of SMA actuator technology. Examples of multiple commercially available geometric forms of SMA actuators are given and the functionalities that they provide are described. This is followed by examples of multiple commercial products incorporating such SMA actuators. Given that there are literally millions of commercial products incorporating conventional actuator technologies, indications are given as to why there are their less than 1000 that utilize SMA. Experience based challenges to the commercial use of SMA actuators are described. Besides having to compete with existing non-SMA technology which is quite mature additional challenges that are unique to SM actuators are indicated these including a wider than expected set of technical engineering problems and challenges and that a broader scope of dynamics is required.

  6. Acupuntura un tratamiento viable para las adicciones en Colombia

    Directory of Open Access Journals (Sweden)

    Hernán López-Suescún

    2013-09-01

    Full Text Available Resumen La acupuntura es una antigua técnica terapéutica desarrollada en China, que ha evidenciado ser efectiva en síntomas como las náuseas, vómito y dolor dentario. A pesar del sustento fisiológico que posibilitaría un uso efectivo en otras patologías, incluyendo el campo de las adicciones, los estudios son contradictorios, posiblemente por la diferencias de visión entre la medicina oriental y la occidental. El consumo de psicoactivos es un problema de salud pública en Colombia y en el mundo que genera grandes costos tangibles e intangibles, los cuales, en países desarrollados, puede llegar hasta el 1,6 % del PIB. En contraste, el beneficio económico del tratamiento de las adicciones, según las Naciones Unidas Contra la Droga y el Delito (UNODC, está entre 1:3 a 1:13; por lo tanto, cualquier esfuerzo que se realice en favor de los consumidores es una ganancia. Con base en estos datos, los organismos internacionales han generado políticas que ayudan a aminorar estos efectos. Colombia, como integrante de estos organismos, ha realizado varios compromisos para llevar a cabo dichas metas. Los tratamientos con auriculoterapia, como el protocolo NADA (National Acupuncture Detoxification Association, son los métodos más usados para las adicciones en el mundo, y aunque no se ha logrado evidenciar su efectividad, por su costo, facilidad y el poco riesgo de efectos adversos se hace viable en un país con pocos recursos económicos como Colombia.

  7. Attempts to improve human ovarian transplantation outcomes of needle-immersed vitrification and slow-freezing by host and graft treatments.

    Science.gov (United States)

    Abir, Ronit; Fisch, Benjamin; Fisher, Noa; Samara, Nivin; Lerer-Serfaty, Galit; Magen, Roei; Herman-Edelstein, Michal; Ben-Haroush, Avi; Stein, Anat; Orvieto, Raoul

    2017-05-01

    To investigate if needle-immersed vitrification or slow-freezing yields better implantation results for human ovarian tissue and which method benefits more when combined with the "improvement protocol" of host melatonin treatment and graft incubation with biological glue + vitamin E + vascular endothelial growth factor-A. Human ovarian tissue was preserved by needle-immersed vitrification or slow-freezing and transplanted into immunodeficient mice, either untreated (groups A and C, respectively) or treated with the improvement protocol (groups B and D, respectively). Grafted and ungrafted slices were evaluated by follicle counts, apoptosis assay and immunohistochemistry for Ki67 and platelet endothelial cell adhesion molecule (PECAM). Follicle number in the recovered grafts was limited. The number of atretic follicles was significantly higher after vitrification with/without the improvement protocol and slow-freezing than that after slow-freezing + the improvement protocol. Stroma cell apoptosis was the lowest in the group D. PECAM staining showed a peripheral and diffuse pattern in the group D (mostly normal follicular morphology) and a diffuse pattern in all other groups (few follicles, mostly atretic), with significantly higher diffuse levels in the vitrification groups. Ki67 staining was identified in all normal follicles. Follicles did not survive transplantation in the vitrification groups. Ovarian sample preparation with slow-freezing + the improvement protocol appears to yield better implantation outcomes than needle-immersed vitrification with/without the improvement protocol. The real quality of frozen tissue can be assessed only after grafting and not after thawing/warming.

  8. Morphological and functional preservation of pre-antral follicles after vitrification of macaque ovarian tissue in a closed system.

    Science.gov (United States)

    Ting, A Y; Yeoman, R R; Campos, J R; Lawson, M S; Mullen, S F; Fahy, G M; Zelinski, M B

    2013-05-01

    What are the appropriate conditions to vitrify the macaque ovarian cortex in a large-volume, closed system that will preserve functional pre-antral follicles? The combination of glycerol, ethylene glycol (EG) and polymers with cooling in liquid nitrogen (LN2) vapor and a two-step warming procedure was able to preserve tissue and follicle morphology as well as function of a small population of secondary follicles in the macaque ovarian cortex following vitrification in a closed system. For prepubertal cancer patients or those who require immediate cancer therapy, ovarian tissue cryopreservation offers the only hope for future fertility. However, the efficacy of live birth from the transplantation of cryopreserved ovarian tissue is still unclear. In addition, live birth from cryopreserved ovarian tissue has only been demonstrated after tissue autotransplantation, which poses the risk of transmitting metastatic cancer cells back to the cancer survivor in certain cancers. Non-human primate model, n = 4, randomized, control versus treatment. End-points were collected from tissue histology, tissue culture (48 h) and isolated secondary follicle culture (6 weeks). Two vitrification solutions (VSs) containing EG + glycerol (VEG) and EG + dimethylsulfoxide (VED) were examined for vitrification, devitrification and thermodynamic properties. Once the optimal VS was determined, macaque ovarian cortical pieces (3 × 3 × 0.5 mm(3)) were divided into fresh and two vitrified groups (VEG and VED). For the vitrification groups, tissues were exposed to 1/4, 1/2 and 1× VS for 5 min/step as well as 1× VS + polymers for 1 min at 37°C, loaded into high-security straws with 1 ml of VS + polymers, heat sealed and cooled in LN2 vapor. Samples were warmed in a 40°C water bath and cryoprotective agents were diluted with 1, 0.5, 0.25 and 0 M sucrose. Tissues were fixed for histological analysis and cultured with bromodeoxyuridine (BrdU). Secondary follicles from VEG tissues were

  9. Forced collapse of the blastocoel cavity improves developmental potential in cryopreserved bovine blastocysts by slow-rate freezing and vitrification.

    Science.gov (United States)

    Min, S-H; Kim, J-W; Lee, Y-H; Park, S-Y; Jeong, P-S; Yeon, J-Y; Park, H; Chang, K-T; Koo, D-B

    2014-08-01

    This study was conducted to evaluate the effectiveness of forced collapse of the blastocoel before slow-rate freezing and vitrification of bovine blastocysts. Cryopreservation of bovine blastocysts has been proposed as a tool to improve the feasibility of cattle production using the embryo transfer technique. However, the low efficiency of frozen-thawed embryos survival and further development is a crucial problem. In this study, bovine in vitro and in vivo blastocysts were slow-rate frozen and vitrified after forced blastocoele collapse (FBC) of the blastocyst cavity by puncturing the blastocoele with a pulled Pasteur pipet. Differences in the developmental potential of frozen-thawed blastocysts derived from FBC and non-FBC groups were found in both slow-rate freezing and vitrification. Furthermore, we found that the total cell number of blastocysts in FBC groups was increased and the index of apoptosis in FBC groups was decreased. Consistent with these results, real-time RT-PCR analysis data showed that expression of the anti-apoptotic Bcl-XL gene was significantly increased by FBC groups, whereas expression of the pro-apoptotic Bax gene was significantly decreased by FBC groups. Our results also showed that pregnancy outcomes in both slow-rate frozen and vitrified bovine in vivo blastocysts could be improved by reducing the fluid content after FBC of the blastocyst cavity. Therefore, we suggest that FBC of the blastocyst cavity with a pulled Pasteur pipet is an effective pre-treatment technique for both slow-rate freezing and vitrification of bovine blastocysts. © 2014 Blackwell Verlag GmbH.

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

    Science.gov (United States)

    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.

  11. Cell counts and survival to vitrification of bovine in vitro produced blastocysts subjected to sublethal high hydrostatic pressure.

    Science.gov (United States)

    Trigal, B; Muñoz, M; Gómez, E; Caamaño, J N; Martin, D; Carrocera, S; Casais, R; Diez, C

    2013-04-01

    This work analyses the effects of a high hydrostatic pressure (HHP) treatment on in vitro survival of in vitro produced (IVP) bovine embryos vitrified with the Cryologic Vitrification Method (CVM). Consequences on embryo quality in terms of cell proliferation and differentiation, and levels of embryonic Heat Shock Protein 70 (Hsp-70) were also examined. Day 7 and 8 bovine in vitro-produced blastocysts were submitted to an HHP treatment (60 MPa, at 32 °C for 1 h) and allowed to recover for 1 or 2 h in culture medium. The HHP treatment did not improve blastocyst survival rates after vitrification/warming. Survival (24 h post-warming) and hatching (48 h post-warming) rates were 79.3 ± 4.9 and 51.8 ± 4.2 vs 73.9 ± 4.2 and 44.7 ± 4.1 for untreated controls and HHP-treated embryos, respectively. Total cell numbers measured in fresh embryos were reduced after 1 h at 32 °C, with or without HHP treatment, indicating that cell proliferation was stopped as a result of stress. Vitrified HHP-treated embryos that hatched at 48 h after warming showed increased cell numbers in their ICM compared with untreated controls (50.2 ± 3.1 vs 38.8 ± 2.7), indicating higher embryo quality. Treatment of blastocysts with HHP did not alter the level of the Hsp-70 protein. In our conditions, HHP treatment did not affect the cryoresistance of these embryos. However, combination of HHP treatment and vitrification in fibreplugs resulted in an increase in the ICM cell number of hatched embryos 48 h post-warming. © 2012 Blackwell Verlag GmbH.

  12. Damage to fetal bovine ovarian tissue caused by cryoprotectant exposure and vitrification is mitigated during tissue culture.

    Science.gov (United States)

    Mouttham, Lara; Fortune, Joanne E; Comizzoli, Pierre

    2015-08-01

    The objective of this study is to characterize the impact of exposure to cryoprotectants followed by vitrification on primordial follicle survival and activation using a fetal bovine model. In the first study, fetal bovine cortical pieces were exposed to cryoprotectants with or without sucrose and cultured up to 7 days in the presence or absence of insulin. In the second study, cortical pieces were exposed to cryoprotectants with or without sucrose, vitrified, and cultured up to 7 days after warming in the presence or absence of insulin. Viability and morphology of follicles, as well as proliferation and/or DNA repair in ovarian tissue were analyzed. When compared to non-exposed controls, normal follicular morphology was affected in groups exposed to cryoprotectants only immediately post-exposure and after 1 day of culture, but improved by day 3 and did not significantly differ by day 7. Similarly, normal follicular morphology was compromised in vitrified groups after warming and on day 1 compared to controls, but improved by days 3 and 7. Proliferation and/or DNA repair in ovarian tissue was not affected by vitrification in this model. Cryoprotectant exposure and vitrification of ovarian tissue did not impair the activation of primordial follicles in response to insulin, although activation was delayed relative to non-exposed controls. Interestingly, sucrose had no noticeable protective effect. Vitrified fetal bovine ovarian tissue has the intrinsic capacity to mitigate the immediate damage to primordial follicles' morphology and retains the capacity to activate. These findings provide a basis for a successful cryopreservation protocol for ovarian cortical tissue in other species including humans.

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

    Science.gov (United States)

    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 Embryo survival (hatching rate) was higher in vitrified versus slow-frozen embryos (43% versus 12%, respectively, P 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 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.

  14. Comparison of different vitrification protocols on viability after transfer of ovine blastocysts in vitro produced and in vivo derived.

    Science.gov (United States)

    Dattena, M; Accardo, C; Pilichi, S; Isachenko, V; Mara, L; Chessa, B; Cappai, P

    2004-08-01

    We compare different vitrification protocols on the pregnancy and lambing rate of in vitro produced (IVP) and in vivo derived (IVD) ovine embryos. Ovine blastocysts were produced by in vitro maturation, fertilization and culture of oocytes collected from slaughtered ewes or superovulated and inseminated animals. Embryos were cryopreserved after exposure at room temperature either for 5 min in 10% glycerol (G), then for 5 min in 10% G + 20% ethylene glycol (EG), then for 30 s in 25% G + 25% EG (glycerol group), or for 3 min in 10% EG + 10% dimethyl sulphoxide (DMSO), then for 30s in 20% EG + 20% DMSO + 0.3 M sucrose (DMSO group). One group of in vitro produced embryos was cryopreserved similarly to the DMSO group, but with 0.75 M sucrose added to the vitrification solution (DMSO 0.75 group). Glycerol group embryos were then loaded into French straws or open pulled Straws (OPS) while the DMSO group embryos were all loaded into OPS and directly plunged into liquid nitrogen. Embryos were warmed with either a one step or three step process. In the one step process, embryos were placed in 0.5 M sucrose. The three-step process was a serial dilution in 0.5, 0.25 and 0.125 M sucrose. The embryos of DMSO 0.75 group were warmed directly by plunging them into tissue culture medium-199 (TCM-199) + 20% foetal bovine serum (FBS) in the absence of sucrose (direct dilution). Following these manipulations, the embryos were transferred in pairs into synchronised recipient ewes and allowed to go to term. The pregnancy and the lambing rate within each group of IVP and IVD embryos indicated that there was no statistical difference among the vitrification protocols.

  15. Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation

    Science.gov (United States)

    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

  16. Sheep Isolated Secondary Follicles Are Able to Produce Metaphase II Oocytes After Vitrification and Long-Term In Vitro Growth.

    Science.gov (United States)

    Lunardi, Franciele Osmarini; de Aguiar, Francisco Leo Nascimento; Apolloni, Livia Brunetti; Duarte, Ana Beatriz Graça; de Sá, Naiza Arcângela Ribeiro; Leal, Érica Suzanne Soares; Sales, Antonia Debora; Lobo, Carlos Henrique; Campello, Cláudio Cabral; Smitz, Johan; Apgar, Gary Allen; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

    2017-08-01

    The vitrification of preantral follicles followed by in vitro growth (IVG) could be valuable to produce fertilizable oocytes. However, the meiotic resumption rates of oocytes cultured from vitrified secondary follicles (SF) have been reported as suboptimal. This study aimed to verify two base media (alpha modification of minimum essential medium, α-MEM, and tissue culture medium 199, TCM199) on vitrified SF regarding different requirements during IVG. Sheep ovarian fragments were divided in six groups: (1) Fresh groups (Control α-MEM and TCM199): SF without vitrification; (2) Follicle-Vitrified (Follicle-Vit α-MEM and TCM199): SF vitrified after isolation; and (3) Tissue-Vitrified (Tissue-Vit α-MEM and TCM199): SF vitrified enclosed in ovarian fragments and, subsequently, isolated. The isolated SF were submitted to IVG for 18 days. Thereafter, the recovered cumulus-oocyte complexes (COCs) underwent in vitro maturation (IVM) and evaluation of chromatin configuration. Follicular granulosa cells were analyzed for their gene expression of Bax, Bcl2, and Connexins (CX) 37 and 43. COCs from in vivo antral follicles were used as in vivo control. Data were analyzed by analysis of variance, Tukey, and chi-square tests. Differences were considered significant if p-value is  0.05) oocyte diameter postmaturation. Oocytes from Follicle-Vit in α-MEM reached metaphase II stage after IVM. Gene expression for CX37, CX43, and Bax was lower in Tissue-Vit groups. For Bcl2, the gene expression was the opposite. In conclusion, during IVG for 18 days, maximal oocyte meiotic resumption was not negatively impacted by vitrification and was greatest for isolated SF using α-MEM as a medium.

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

    Energy Technology Data Exchange (ETDEWEB)

    SCHAUS, P.S.

    2006-07-21

    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 Waste 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.

  18. Vitrification of ICSI- and IVF-derived bovine blastocysts by minimum volume cooling procedure: effect of developmental stage and age

    OpenAIRE

    Abdalla, H.; Shimoda, M.; Hara, H.; Morita, H.; Kuwayama, M; Hirabayashi, M.; Hochi, S

    2010-01-01

    The objective was to investigate the effects of developmental stage (fully-expanded or expanding blastocysts) and/or age (harvested on Days 7 or 8) on post-vitrification in vitro survival of bovine blastocysts derived from intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Post-warming survival (re-expansion of blastocoele within 24 h) of ICSI-derived fully-expanded blastocysts (80%) was similar to that of their IVF-derived counterparts (88%). However, the ability of ICS...

  19. Coscheduling in Clusters: Is It a Viable Alternative?

    Energy Technology Data Exchange (ETDEWEB)

    Choi, G S; Kim, J H; Ersoz, D; Yoo, A B; Das, C R

    2003-11-10

    than spin-based techniques like PB on a Linux platform. Third, the proposed HYBRID scheduling provides the best performance-energy behavior and can be implemented on any cluster with little effort. All these results suggest that blocking-based coscheduling techniques are viable candidates to be used instead of batching scheme for significant performance-energy benefits.

  20. Vitrification of noble metals containing NCAW simulant with an engineering scale melter (ESM): Campaign report

    Energy Technology Data Exchange (ETDEWEB)

    Grunewald, W.; Roth, G.; Tobie, W.; Weisenburger, S.; Weiss, K.; Elliott, M.; Eyler, L.L.

    1996-03-01

    ESM has been designed as a 10th-scale model of the DWPF-type melter, currently the reference melter for nitrification of Hanford double shell tankwaste. ESM and related equipment have been integrated to the existing mockup vitrification plant VA-WAK at KfK. On June 2-July 10, 1992, a shakedown test using 2.61 m{sup 3} of NCAW (neutralized current acid waste) simulant without noble metals was performed. On July 11-Aug. 30, 1992, 14.23 m{sup 3} of the same simulant with nominal concentrations of Ru, Rh, and Pd were vitrified. Objective was to investigate the behavior of such a melter with respect to discharge of noble metals with routine glass pouring via glass overflow. Results indicate an accumulation of noble metals in the bottom area of the flat-bottomed ESM. About 65 wt% of the noble metals fed to the melter could be drained out, whereas 35 wt% accumulated in the melter, based on analysis of glass samples from glass pouring stream in to the canisters. After the melter was drained at the end of the campaign through a bottom drain valve, glass samples were taken from the residual bottom layer. The samples had significantly increased noble metals content (factor of 20-45 to target loading). They showed also a significant decrease of the specific electric resistance compared to bulk glass (factor of 10). A decrease of 10- 15% of the resistance between he power electrodes could be seen at the run end, but the total amount of noble metals accumulated was not yet sufficient enough to disturb the Joule heating of the glass tank severely.

  1. Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes.

    Directory of Open Access Journals (Sweden)

    Giulia Rusciano

    Full Text Available Although oocyte cryopreservation has great potentials in the field of reproductive technologies, it still is an open challenge in the majority of domestic animals and little is known on the biochemical transformation induced by this process in the different cellular compartments. Raman micro-spectroscopy allows the non-invasive evaluation of the molecular composition of cells, based on the inelastic scattering of laser photons by vibrating molecules. The aim of this work was to assess the biochemical modifications of both the zona pellucida and cytoplasm of vitrified/warmed in vitro matured bovine oocytes at different post-warming times. By taking advantage of Principal Component Analysis, we were able to shed light on the biochemical transformation induced by the cryogenic treatment, also pointing out the specific role of cryoprotective agents (CPs. Our results suggest that vitrification induces a transformation of the protein secondary structure from the α-helices to the β-sheet form, while lipids tend to assume a more packed configuration in the zona pellucida. Both modifications result in a mechanical hardening of this cellular compartment, which could account for the reduced fertility rates of vitrified oocytes. Furthermore, biochemical modifications were observed at the cytoplasmic level in the protein secondary structure, with α-helices loss, suggesting cold protein denaturation. In addition, a decrease of lipid unsaturation was found in vitrified oocytes, suggesting oxidative damages. Interestingly, most modifications were not observed in oocytes exposed to CPs, suggesting that they do not severely affect the biochemical architecture of the oocyte. Nevertheless, in oocytes exposed to CPs decreased developmental competence and increased reactive oxygen species production were observed compared to the control. A more severe reduction of cleavage and blastocyst rates after in vitro fertilization was obtained from vitrified oocytes. Our

  2. Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes.

    Science.gov (United States)

    Rusciano, Giulia; De Canditiis, Carolina; Zito, Gianluigi; Rubessa, Marcello; Roca, Maria Serena; Carotenuto, Rosa; Sasso, Antonio; Gasparrini, Bianca

    2017-01-01

    Although oocyte cryopreservation has great potentials in the field of reproductive technologies, it still is an open challenge in the majority of domestic animals and little is known on the biochemical transformation induced by this process in the different cellular compartments. Raman micro-spectroscopy allows the non-invasive evaluation of the molecular composition of cells, based on the inelastic scattering of laser photons by vibrating molecules. The aim of this work was to assess the biochemical modifications of both the zona pellucida and cytoplasm of vitrified/warmed in vitro matured bovine oocytes at different post-warming times. By taking advantage of Principal Component Analysis, we were able to shed light on the biochemical transformation induced by the cryogenic treatment, also pointing out the specific role of cryoprotective agents (CPs). Our results suggest that vitrification induces a transformation of the protein secondary structure from the α-helices to the β-sheet form, while lipids tend to assume a more packed configuration in the zona pellucida. Both modifications result in a mechanical hardening of this cellular compartment, which could account for the reduced fertility rates of vitrified oocytes. Furthermore, biochemical modifications were observed at the cytoplasmic level in the protein secondary structure, with α-helices loss, suggesting cold protein denaturation. In addition, a decrease of lipid unsaturation was found in vitrified oocytes, suggesting oxidative damages. Interestingly, most modifications were not observed in oocytes exposed to CPs, suggesting that they do not severely affect the biochemical architecture of the oocyte. Nevertheless, in oocytes exposed to CPs decreased developmental competence and increased reactive oxygen species production were observed compared to the control. A more severe reduction of cleavage and blastocyst rates after in vitro fertilization was obtained from vitrified oocytes. Our experimental

  3. Oocyte pre-IVM with caffeine improves bovine embryo survival after vitrification.

    Science.gov (United States)

    Bernal-Ulloa, Sandra Milena; Lucas-Hahn, Andrea; Herrmann, Doris; Hadeler, Klaus-Gerd; Aldag, Patrick; Baulain, Ulrich; Niemann, Heiner

    2016-09-15

    Cryopreservation of in vitro produced bovine embryos is associated with significantly reduced survival rates, mainly due to insufficient quality of the embryos. Caffeine supplementation during IVM has been used to delay meiotic resumption and concomitantly also increased embryo quality. Here, we investigated the influence of pre-IVM with caffeine on oocyte maturation, intraoocyte cAMP concentration, developmental competence after IVF, and blastocyst cryotolerance. Oocytes were obtained by slicing of ovaries and were submitted to either 2 hours culture before IVM with or without caffeine (0, 1, 5, 10, 20, 30 mM), or standard IVM (no pre-IVM). Oocytes were in vitro matured and fertilized and zygotes were cultured under standard in vitro conditions until Day 8. Expanded blastocysts derived from either standard control or the 10-mM caffeine treatments were submitted to vitrification. Caffeine delayed meiotic resumption after 9-hour IVM in a concentration-dependent manner. The cAMP levels were similar before and after IVM. Matured oocytes, cleavage, and blastocyst rates were reduced in the 30-mM caffeine concentration and were similar among the other treatment groups. Number and proportion of inner cell mass and trophectoderm cells in blastocysts did not differ among treatments. Forty-eight hours after thawing, hatching rates were higher in the 10-mM caffeine group (73.8%) compared with the standard control (59.7%). Reexpansion rates and total number of cells after 48 hours were similar in both treatments. The ratio of live/total cells was higher in the caffeine treatment. These results suggest that caffeine supplementation before IVM delayed meiotic resumption and improved blastocyst quality shown in higher cryotolerance. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Vitrification of incinerated tannery sludge in silicate matrices for chromium stabilization.

    Science.gov (United States)

    Varitis, S; Kavouras, P; Pavlidou, E; Pantazopoulou, E; Vourlias, G; Chrissafis, K; Zouboulis, A I; Karakostas, Th; Komninou, Ph

    2017-01-01

    The vitrification process was applied for the stabilization and solidification of a rich in chromium ash that was the by-product of incineration of tannery sludge. Six different batch compositions were produced, based on silica as the glass former and sodium and calcium oxides as flux agents. As-vitrified products (monoliths) were either composed of silicate matrices with separated from the melt Eskolaite (Cr2O3) crystallites or were homogeneous glasses (in one case). All as-vitrified products were thermally treated in order to transform them to partially crystallized, i.e. devitrified products. Devitrification is an important part of the work since studying the transformation of the initial as-vitrified products into glass-ceramics with better properties could result to stabilized products with potential added value. The devitrified products were diversified by the effective crystallization mode and separated crystal phase composition. These variations originated from differences in: (a) batch composition of the initial as-vitrified products and (b) thermal treatment conditions. In devitrified products crystallization led to the separation of Devitrite (Na2Ca3Si6O16), Combeite (Na4Ca4Si6O18) and Wollastonite (CaSiO3) crystalline phases, while Eskolaite crystallites were not affected by thermal treatment. Leaching test results revealed that chromium was successfully stabilized inside the as-vitrified monoliths. Devitrification impairs chromium stabilization, only in the case where the initial as-vitrified product was a homogeneous glass. In all other cases, devitrification did not affect successful chromium stabilization. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Isothermal vitrification methodology development for non-cryogenic storage of archival human sera.

    Science.gov (United States)

    Less, Rebekah; Boylan, Kristin L M; Skubitz, Amy P N; Aksan, Alptekin

    2013-04-01

    Biorepositories worldwide collect human serum samples and store them for future research. Currently, hundreds of biorepositories across the world store human serum samples in refrigerators, freezers, or liquid nitrogen without following any specific cryopreservation protocol. This method of storage is both expensive and potentially detrimental to the biospecimens. To decrease both cost of storage and the freeze/thaw stresses, we explored the feasibility of storing archival human serum samples at non-cryogenic temperatures using isothermal vitrification. When biospecimens are vitrified, biochemical reactions can be stopped, the specimen ceases to degrade, and macromolecules can be stabilized without requiring cryogenic storage. In this study, 0.2, 0.4, or 0.8M trehalose; 0, 0.005 or 0.01M dextran; and 0 or 10% (v/v) glycerol was added to human serum samples. The samples were either dried diffusively as sessile droplets or desiccated under vacuum after they are adsorbed onto glass microfiber filters. The glass transition temperatures (Tg) of the desiccated samples were measured by temperature-ramp Fourier Transform Infrared (FTIR) spectroscopy. Sera samples vitrified at 4±2°C when 0.8M trehalose and 0.01M dextran were added and the samples were vacuum dried for two hours. Western immunoblotting showed that vitrified serum proteins were minimally degraded when stored for up to one month at 4°C. About 80% of all proteins were recovered after storage at 4°C on glass microfiber filters, and recovery did not decrease with storage time. These results demonstrated the feasibility of long-term storage of vitrified serum at hypothermic (and non-cryogenic) temperatures. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Cryopreservation of seeds of Encholirium spectabile Martius ex Schultes f. by the vitrification method

    Directory of Open Access Journals (Sweden)

    Edilene Aparecida Preti Ferrari

    Full Text Available ABSTRACT The bromeliad Encholirium spectabile Martius ex Schultes f. is a endemic species of the Caatinga. The aim of this study was to evaluate the efficiencies of cryoprotectant in bromeliad seeds. The treatments consisted of immersing seeds in different cryoprotectant solutions and vitrification solutions before immersion in liquid nitrogen (NL at -196 °C, as the following treatments: T1- Control: without cryoprotectants; T2- glycerol 2M (20 min + PVS2 (10 min, T3- glycerol 2M (20 min + PVS2 with phloroglucinol 1% (10 min, T4- sucrose 0.4M (20 min + PVS2 (10 min; T5- sucrose 0.4M (20 min + PVS2 with phloroglucinol 1% (10 min; T6- glycerol 2M (20 min + sucrose 0.4M (20 min + PVS2 (10 min, T7- glycerol 2M (20 min + sucrose 0.4M (20 min + PVS2 with phloroglucinol 1% (10 min. The experimental design was a completely randomized design with seven treatments and five replicates per treatment. Data were subjected to analysis of variance (ANOVA and means were compared by the Tukey test at 5%. The seed moisture at the beginning of the experiment was 8.4%. There were statistically significant for the variables percentage of seeds germinated and seedling length, but for the variable seedling dry weight, T3 treatment was presented a value statistically high to T1, T4, T5 and T6, but did not differ from T2 and T7 treatments. It is concluded that for the cryopreservation of seeds of Encholirium spectabile is not necessary to use cryoprotectant solutions.

  7. In situ vitrification of Oak Ridge National Laboratory soil and limestone

    Energy Technology Data Exchange (ETDEWEB)

    Carter, J.G.; Bates, S.O.; Maupin, G.D.

    1987-03-01

    Process feasibility studies were successfully performed on two different developmental scales to determine the technical application of in situ vitrification (ISV) to Oak Ridge National Laboratory (ORNL) intermediate-level waste. In the laboratory, testing was performed on crucibles containing quantities of 50% ORNL soil and 50% ORNL limestone. In the engineering-scale testing, a 1/12-scaled simulation of ORNL Trench 7 was constructed and vitrified, resulting in waste product soil and limestone concentrations of 68% and 32%, respectively. Results from the two scales of testing indicate that the ORNL intermediate-level waste sites may be successfully processed by ISV; the waste form will retain significant quantities of the cesium and strontium. Because /sup 137/Cs is the major component of the radionuclide inventory in the ORNL seepage pits and trenches, final field process decontamination factors (i.e., off gas at the ground surface relative to the waste inventory) of 10/sup 4/ are desired to minimize activity buildup in the off-gas system. These values were realized during the engineering-scale test for both cesium and strontium. The vitrified material effectively contained 99.996% of the cesium and strontium placed in the engineering-scale test. This is equivalent to decontamination factors of greater than 10/sup 4/. Volume reduction for the engineering-scale test was 60%. No migration of the cesium to the uncontaminated surrounding soil was detected. These favorable results indicate that, once verified in a pilot-scale test, an adequately designed ISV system could be produced to treat the ORNL seepage pits and trenches without excessive activity accumulation in the off-gas treatment system.

  8. Investigation of potential analytical methods for redox control of the vitrification process. [Moessbauer

    Energy Technology Data Exchange (ETDEWEB)

    Goldman, D.S.

    1985-11-01

    An investigation was conducted to evaluate several analytical techniques to measure ferrous/ferric ratios in simulated and radioactive nuclear waste glasses for eventual redox control of the vitrification process. Redox control will minimize the melt foaming that occurs under highly oxidizing conditions and the metal precipitation that occurs under highly reducing conditions. The analytical method selected must have a rapid response for production problems with minimal complexity and analyst involvement. The wet-chemistry, Moessbauer spectroscopy, glass color analysis, and ion chromatography techniques were explored, with particular emphasis being placed on the Moessbauer technique. In general, all of these methods can be used for nonradioactive samples. The Moessbauer method can readily analyze glasses containing uranium and thorium. A shielded container was designed and built to analyze fully radioactive glasses with the Moessbauer spectrometer in a hot cell environment. However, analyses conducted with radioactive waste glasses containing /sup 90/Sr and /sup 137/Cs were unsuccessful, presumably due to background radiation problems caused by the samples. The color of glass powder can be used to analyze the ferrous/ferric ratio for low chromium glasses, but this method may not be as precise as the others. Ion chromatography was only tested on nonradioactive glasses, but this technique appears to have the required precision due to its analysis of both Fe/sup +2/ and Fe/sup +3/ and its anticipated adaptability for radioactivity samples. This development would be similar to procedures already in use for shielded inductively coupled plasma emission (ICP) spectrometry. Development of the ion chromatography method is therefore recommended; conventional wet-chemistry is recommended as a backup procedure.

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

    Directory of Open Access Journals (Sweden)

    Alisha Gupta

    Full Text Available Cryopreservation is known for its marked deleterious effects on embryonic health. Bovine compact morulae were vitrified or slow-frozen, and post-warm morulae were cultured to the expanded blastocyst stage. Blastocysts developed from vitrified and slow-frozen morulae were subjected to microarray analysis and compared with blastocysts developed from unfrozen control morulae for differential gene expression. Morula to blastocyst conversion rate was higher (P < 0.05 in control (72% and vitrified (77% than in slow-frozen (34% morulae. Total 20 genes were upregulated and 44 genes were downregulated in blastocysts developed from vitrified morulae (fold change ≥ ± 2, P < 0.05 in comparison with blastocysts developed from control morulae. In blastocysts developed from slow-frozen morulae, 102 genes were upregulated and 63 genes were downregulated (fold change ≥ ± 1.5, P < 0.05. Blastocysts developed from vitrified morulae exhibited significant changes in gene expression mainly involving embryo implantation (PTGS2, CALB1, lipid peroxidation and reactive oxygen species generation (HSD3B1, AKR1B1, APOA1 and cell differentiation (KRT19, CLDN23. However, blastocysts developed from slow-frozen morulae showed changes in the expression of genes related to cell signaling (SPP1, cell structure and differentiation (DCLK2, JAM2 and VIM, and lipid metabolism (PLA2R1 and SMPD3. In silico comparison between blastocysts developed form vitrified and slow-frozen morulae revealed similar changes in gene expression as between blastocysts developed from vitrified and control morulae. In conclusion, blastocysts developed form vitrified morulae demonstrated better post-warming survival than blastocysts developed from slow-frozen morulae but their gene expression related to lipid metabolism, steroidogenesis, cell differentiation and placentation changed significantly (≥ 2 fold. Slow freezing method killed more morulae than vitrification but those which survived up to

  10. Viabilidade e fertilização in vitro de oócitos bovinos após vitrificação Viability and in vitro fertilization of bovine oocytes after vitrification

    Directory of Open Access Journals (Sweden)

    Sérgio Galbinski

    2003-09-01

    ários para proteção da zona pelúcida e do oolema.PURPOSE: to verify vitrification techniques using 6 M DMSO to cryopreserve in vitro matured bovine oocytes, and to assess the effects of the time of exposure to vitrification solutions (VS. METHODS: dilutions of VS were prepared from the stock VS (VS 100% consisting of 6 M DMSO to give 25 and 65% DMSO solutions. Bovine oocytes were in vitro matured for 18-22 h. Matured oocytes were placed first into 25% VS, at room temperature for 5 min, then transferred to 65% VS, before being pipetted into the 100% VS in plastic straws. Three experimental groups were formed: in the first group, time of pipetting through 65% VS and loading the straw took up to 60 s, in the second group it did not exceed 30 s. For thawing, straws were held in air for 10 s and then in a water bath for 10 s. The contents of each straw were expelled in sucrose solution and held for 5 min. In the third experimental group, oocytes went through all VS, but were not vitrified. All retrieved oocytes were inseminated. For control, fresh, in vitro matured oocytes were inseminated. RESULTS: after vitrification, 69.1 and 59.8% of the oocytes were retrieved from the 30 s and 60 s groups, respectively, and 93 and 89% of these oocytes appeared morphologically normal 24 h after insemination, respectively. In the group of oocytes exposed without vitrification, 75.6% were retrieved and 84.7% were morphologically viable, 24 h after insemination. No fertilization was observed in the experimental groups. Among controls, 65.4% were fertilized. CONCLUSIONS: the vitrification technique using 6 M DMSO is not a feasible approach to cryopreserve in vitro matured bovine oocytes. Decreasing the time of exposure to VS did not overcome deleterious effects of the procedure on the fertilizability of oocytes. Improvements in the technique are needed to protect the zona pellucida and oolemma.

  11. Effect of myocardial revascularisation on left ventricular systolic function in patients with and without viable myocardium: should non-viable segments be revascularised?

    Science.gov (United States)

    Stipac, Alja Vlahovic; Stankovic, Ivan; Vidakovic, Radosav; Putnikovic, Biljana; Ilic, Ivan; Milicic, Biljana; Neskovic, Aleksandar N

    2013-12-01

    To assess the effect of surgical revascularisation on left ventricular (LV) systolic function in patients with viable and non-viable dysfunctional LV segments determined by low dose dobutamine stress echocardiography (DSE). Prospective observational cohort study. Single tertiary care centre. Consecutive patients referred to surgical revascularisation (n=115). DSE and surgical revascularisation. Functional recovery defined as increase in ejection fraction ≥ 5% 1 year after revascularisation in patients with and without viable myocardium (viability defined as improvement of contractility in ≥ 4 LV segments on DSE). The mean age, ejection fraction and wall motion score index (WMSi) of patients were 59 ± 9 years, 44 ± 9% and 1.82 ± 0.31, respectively. There was no difference between DSE positive and DSE negative patients for any of those parameters at baseline study (p>0.05 for all). After 12 months, the ejection fraction increased 11 ± 1% in patients with viable myocardium vs 7 ± 1% in patients without viable myocardium (p=0.002). Moreover, in patients with viable myocardium, the greatest increase of ejection fraction occurred 1 month after surgery (9 ± 1%), whereas in those patients with negative DSE the ejection fraction increased more gradually (2±1% after 1 month, p=0.002 between groups for 1 month vs preoperative value), but still improved after 12 months follow-up (pmyocardial revascularisation. Functional recovery continuously occurs throughout the first year after surgical treatment.

  12. A technique for determining viable military logistics support alternatives

    Science.gov (United States)

    Hester, Jesse Stuart

    A look at today's US military will see them operating much beyond the scope of protecting and defending the United States. These operations now consist of, but are not limited to humanitarian aid, disaster relief, peace keeping, and conflict resolution. This broad spectrum of operational environments has necessitated a transformation of the individual military services to a hybrid force that is attempting to leverage the inherent and emerging capabilities and strengths of all those under the umbrella of the Department of Defense (DOD), this concept has been coined Joint Operations. Supporting Joint Operations requires a new approach to determining a viable military logistics support system. The logistics architecture for these operations has to accommodate scale, time, varied mission objectives, and imperfect information. Compounding the problem is the human in the loop (HITL) decision maker (DM) who is a necessary component for quickly assessing and planning logistics support activities. Past outcomes are not necessarily good indicators of future results, but they can provide a reasonable starting point for planning and prediction of specific needs for future requirements. Adequately forecasting the necessary logistical support structure and commodities needed for any resource intensive environment has progressed well beyond stable demand assumptions to one in which dynamic and nonlinear environments can be captured with some degree of fidelity and accuracy. While these advances are important, a holistic approach that allows exploration of the operational environment or design space does not exist to guide the military logistician in a methodical way to support military forecasting activities. To bridge this capability gap, a method called Adaptive Technique for Logistics Architecture Solutions (ATLAS) has been developed. This method provides a process that facilitates the use of techniques and tools that filter and provide relevant information to the DM. By doing

  13. Survival and developmental competence of bovine embryos at different developmental stages and separated blastomeres after vitrification in different solutions.

    Science.gov (United States)

    Juanpanich, Theesit; Suttirojpattana, Tayita; Takayama, Mari; Liang, Yuanyuan; Dochi, Osamu; Parnpai, Rangsun; Imai, Kei

    2018-01-01

    Generating techniques to enhance the success of blastomere separation is important for bovine economy, because it increases the number of transferable embryos. This study aimed to identify the optimum cryoprotectants for the vitrification of bovine embryos and the separation of blastomeres at different stages. In experiment 1, expanded blastocysts were vitrified in two different vitrification solutions, either (1) ethylene glycol (EG) + propylene glycol (PG) or (2) EG. The survival rate of blastocysts in the EG + PG was higher than that of the EG. In experiment 2, intact two-cell and eight-cell stage embryos were vitrified in the same solutions used in experiment 1. The EG + PG produced more dead embryos than the EG (P < 0.05). In the EG, the rate of blastocyst formation was similar for the vitrified two- and eight-cell embryos and the non-vitrified ywo-cell embryos. In experiment 3, separated blastomeres of two- and eight-cell embryos were vitrified in EG. There was no difference in the rate of blastocyst formation and total number of cells between the two vitrified groups. In summary, at the blastocyst stage, EG + PG was superior, based on both survival rates and cell numbers; however, at the 2-8 cell stage, the use of EG alone was better than the other groups. © 2017 Japanese Society of Animal Science.

  14. Cytochalasin B efficiency in the cryopreservation of immature bovine oocytes by Cryotop and solid surface vitrification methods.

    Science.gov (United States)

    Sripunya, Nucharin; Liang, Yuanyuan; Panyawai, Kanchana; Srirattana, Kanokwan; Ngernsoungnern, Apichart; Ngernsoungnern, Piyada; Ketudat-Cairns, Mariena; Parnpai, Rangsun

    2014-12-01

    The present study was undertaken to compare the efficacies of Cryotop (CT), solid surface vitrification (SSV) methods and cytochalasin B (CB) treatment for the cryopreservation of immature bovine oocytes, in terms of survival, nuclear maturation, and in vitro development. Solution exposed oocytes were in vitro maturated and fertilized. No difference was found in the rates of survival, nuclear maturation and blastocyst among solution exposed groups and fresh control group, except blastocysts rates in oocytes exposed to CB, cryoprotectant (CPA) and fluorescein diacetate (FDA) group (CB-CPA-FDA) (23%) significantly lower than that of control group (32%). CB pretreated ((+)CB) or non-pretreated ((-)CB) COCs were vitrified either by SSV or CT. Among four vitrified groups the nuclear maturation rates (CT(-)CB: 58%, CT(+)CB: 57%, SSV(-)CB: 60%, SSV(+)CB: 63%), cleavage (CT(-)CB: 36%, CT(+)CB: 24%, SSV(-)CB: 34%, SSV(+)CB: 26%) and blastocysts rates (CT(-)CB: 6%, CT(+)CB: 7%, SSV(-)CB: 4%, SSV(+)CB: 6%) did not differ, but the rates of the four vitrified groups were significantly lower than those of non-vitrified group (81%, 71% and 26%, respectively). We thus conclude that CT and SSV perform equally in vitrification of bovine immature oocytes, and CB did not increase the viability, nuclear maturation, or in vitro development of vitrified oocytes. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Effect of container, vitrification volume and warming solution on cryosurvival of in vitro-produced bovine embryos.

    Science.gov (United States)

    Rios, G L; Mucci, N C; Kaiser, G G; Alberio, R H

    2010-03-01

    The aim of the present research was to develop a low cost and easy to perform vitrification method for in vitro-produced cattle embryos. Effect of container material was evaluated (plastic straw compared to glass capillary, experiment 1), two volume sample (1 compared to 0.5 microL, experiment 2) and warming solution composition medium (Tissue Culture Medium 199 (TCM-199) compared to phosphate buffered saline (PBS), experiment 3) as modifications of the open pulled straw (OPS) system in order to reduce embryo damage caused by exposure to cold. In all experiments, day 7 and expanded blastocysts of cattle were exposed to the vitrification solution 1 for 3 min and 30s in solution 2. After this, embryos were placed in a droplet and loaded in a narrow end container, and immediately submerged into liquid nitrogen. For warming, vitrified embryos were plunged into warming solution 1 for 3 min, and transferred into warming solution 2 for 1 min. Fresh embryos kept in culture were used as control group. Hatching rates were recorded in all cases at day 13. In experiment 1 there was no significant effect of container material on hatching rates. Postwarming survival rate of vitrified embryos was lower than control (27.5% plastic straws, 18.9% glass capillary and 80.5% control, Pstraw (OPS) procedure, and that PBS can replace TCM-199 in warming solutions, but lesser hatching rates should be expected.

  16. ROS-INDUCED OXIDATIVE STRESS IN NOBILE-TYPE DENDROBIUM PROTOCORM-LIKE BODIES (PLBS) DURING VITRIFICATION.

    Science.gov (United States)

    Jia, M; Di, W; Liu, Yan; Shi, Yin; Xie, Y

    Oxidative stress involved in cryopreservation protocols may be responsible for the poor survival of tissues after cryopreservation. In the current study, we aimed to clarify the role of oxidative stress and its relationship with survival rate during cryopreservation of PLBs from nobile-type Dendrobium. ROS, antioxidants and oxidative products and the survival rate in PLBs from Dendrobium Hamanal Lake Dream were determined during vitrification. Relative survival of PLBs decreased significantly after preculture and rewarming (P<0.01). Generation of ·O2(-) and protein carbonyl (PCO) increased significantly after preculture. Dramatic increases in ·O2(-), H2O2 and MDA, and significant decreases in AsA content, activities of SOD and CAT were observed after rewarming. ROS-induced oxidative stress was associated with the poor survival of PLBs following vitrification. ·O2(-) was the predominant ROS resulting in the decreased survival after preculture, while H2O2 together with ·O2(-) appear to be responsible for the survival decrease after rewarming.

  17. The potential for modification in cloning and vitrification technology to enhance genetic progress in beef cattle in Northern Australia.

    Science.gov (United States)

    Taylor-Robinson, Andrew W; Walton, Simon; Swain, David L; Walsh, Kerry B; Vajta, Gábor

    2014-08-01

    Recent advances in embryology and related research offer considerable possibilities to accelerate genetic improvement in cattle breeding. Such progress includes optimization and standardization of laboratory embryo production (in vitro fertilization - IVF), introduction of a highly efficient method for cryopreservation (vitrification), and dramatic improvement in the efficiency of somatic cell nuclear transfer (cloning) in terms of required effort, cost, and overall outcome. Handmade cloning (HMC), a simplified version of somatic cell nuclear transfer, offers the potential for relatively easy and low-cost production of clones. A potentially modified method of vitrification used at a centrally located laboratory facility could result in cloned offspring that are economically competitive with elite animals produced by more traditional means. Apart from routine legal and intellectual property issues, the main obstacle that hampers rapid uptake of these technologies by the beef cattle industry is a lack of confidence from scientific and commercial sources. Once stakeholder support is increased, the combined application of these methods makes a rapid advance toward desirable traits (rapid growth, high-quality beef, optimized reproductive performance) a realistic goal. The potential impact of these technologies on genetic advancement in beef cattle herds in which improvement of stock is sought, such as in northern Australia, is hard to overestimate. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  18. PRODUCTIVITY AND CYTOGENETIC STABILITY OF PROTOCORM-LIKE BODIES OF HYBRID CYMBIDIUM CRYOPRESERVED BY ENCAPSULATION-DEHYDRATION AND VITRIFICATION

    Directory of Open Access Journals (Sweden)

    Jaime A. TEIXEIRA DA SILVA

    2014-12-01

    Full Text Available Transformed and untransformed protocorm-like bodies (PLBs of hybrid Cymbidium Twilight Moon ‘Day Light’ were cryopreserved using two methods (encapsulation-dehydration and vitrification to form new PLBs (neo-PLBs on Teixeira Cymbidium (TC medium. The organogenic response was quantified and genetic stability was assessed by flow cytometry. Intact PLBs produced significantly more neo-PLBs and fresh weight of neo-PLBs than half-PLBs or PLB longitudinal thin cell layers (lTCLs, for two vitrification protocols and for an encapsulation-dehydration method. The addition of 2% sucrose significantly improved the number of neo-PLBs than half-PLBs or PLB lTCLs but any other concentration of sucrose, and all other osmotic agents (mannose, PEG-6000, DMSO at any concentration resulted in significantly worse neo-PLB formation. However, the length of exposure to sucrose did not significantly affect the number or fresh weight of neo-PLBs. The desiccation (air current or silica gel method applied negatively affected the number and fresh weight of neo-PLBs. The length of the cryopreservation period negatively impacted the number of neo-PLBs and fresh weight of transformed or untransformed neo-PLBs. Most severe negative effects were registered after one year of cryopreservation. Cryopreserved neo-PLBs showed high levels of endopolyploidy (8C-64C relative to non-cryopreserved PLBs.

  19. Vitrification of Tokuyasu-style immuno-labelled sections for correlative cryo light microscopy and cryo electron tomography.

    Science.gov (United States)

    Bos, Erik; Hussaarts, Leonie; van Weering, Jan R T; Ellisman, Mark H; de Wit, Heidi; Koster, Abraham J

    2014-05-01

    We present an approach for the preparation of immuno-labelled ultrathin sections from cells or tissue that are compatible with both fluorescence and transmission electron microscopy. Our approach is inspired by a method of Sabanay et al. (1991) that is based on the Tokuyasu technique for immunogold labelling of sections from aldehyde-fixed samples. The difference of this method with the original Tokuyasu technique is that the immuno-labelled sections are stabilized in a thin layer of vitreous water by plunge-freezing prior to electron microscopical observation. The vitrification step allows for phase contrast-based imaging at cryogenic conditions. We show that this immuno-labelling method is well-suited for imaging cellular ultrastructure in three dimensions (tomography) at cryogenic conditions, and that fluorescence associated with the sections is retained. This method is a valuable tool for Correlative Light and Electron Microscopy (CLEM), and we refer to this method in combination with CLEM as VOS (vitrification of sections). We provide examples for the application of VOS using dendritic cells and neurons, and show specifically that this method enables the researcher to navigate to lysosomes and synapses. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Grazing of particle-associated bacteria-an elimination of the non-viable fraction.

    Science.gov (United States)

    Gonsalves, Maria-Judith; Fernandes, Sheryl Oliveira; Priya, Madasamy Lakshmi; LokaBharathi, Ponnapakkam Adikesavan

    Quantification of bacteria being grazed by microzooplankton is gaining importance since they serve as energy subsidies for higher trophic levels which consequently influence fish production. Hence, grazing pressure on viable and non-viable fraction of free and particle-associated bacteria in a tropical estuary controlled mainly by protist grazers was estimated using the seawater dilution technique. In vitro incubations over a period of 42h showed that at the end of 24h, growth coefficient (k) of particle-associated bacteria was 9 times higher at 0.546 than that of free forms. Further, 'k' value of viable cells on particles was double that of free forms at 0.016 and 0.007, respectively. While bacteria associated with particles were grazed (coefficient of removal (g)=0.564), the free forms were relatively less grazed indicating that particle-associated bacteria were exposed to grazers in these waters. Among the viable and non-viable forms, 'g' of non-viable fraction (particle-associated bacteria=0.615, Free=0.0086) was much greater than the viable fraction (particle-associated bacteria=0.056, Free=0.068). Thus, grazing on viable cells was relatively low in both the free and attached states. These observations suggest that non-viable forms of particle-associated bacteria were more prone to grazing and were weeded out leaving the viable cells to replenish the bacterial standing stock. Particle colonization could thus be a temporary refuge for the "persistent variants" where the viable fraction multiply and release their progeny. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  1. Vitrification of in vitro matured oocytes collected from surplus ovarian medulla tissue resulting from fertility preservation of ovarian cortex tissue

    DEFF Research Database (Denmark)

    Yin, Huiqun; Jiang, Hong; Kristensen, Stine Gry

    2016-01-01

    PURPOSE: The aim of the study was to investigate the maturation rate of immature oocytes collected from ovarian medulla tissue normally discarded during preparation of ovarian cortical tissue for fertility preservation. Further we evaluated survival of derived MII oocytes following vitrification ...

  2. OPEN PULLED STRAW (OPS) VITRIFICATION OF PORCINE BLASTOCYSTS: SIMPLE PROCEDURE YIELDING EXCELLENT IN VITRO SURVIVAL, BUT SO FAR NO PIGLETS FOLLOWING TRANSFER

    DEFF Research Database (Denmark)

    Holm, P; Vajta, G; Macháty, Z

    1999-01-01

    In 2 trials, 346 porcine in vivo blastocysts were vitrified in heat-softened and pulled French straws by the Open Pulled Straw (OPS) vitrification method. After thawing, embryos were either cultured in vitro (n=89) for 24 to 48 hours or transferred surgically (n=257) to 11 synchronised recipients...

  3. The influence of polyvinylpyrrolidone (PVP) on the toxicity of cryoprotective agents in different liver cell types and on vitrification outcome in precision-cut liver and kidney slices

    NARCIS (Netherlands)

    Guan, Na; Blomsma, Sylvia; de Jager, Marina; Fahy, Gregory M.; Groothuis, Genoveva; de Graaf, Inge

    2012-01-01

    Cryopreservation of precision-cut tissue slices would strongly improve their application possibilities in ADME-tox studies. Low toxic mixtures of cryoprotectant agents (CPAs) are needed for cryopreservation of cells, and tissues by vitrification. Many of these mixtures contain polyvinylpyrrolidone

  4. Cybernetically sound organizational structures II: Relating de Sitter's design theory to Beer's viable system model

    NARCIS (Netherlands)

    Achterbergh, J.M.I.M.; Vriens, D.J.

    2011-01-01

    - Purpose – The purpose of this paper is to show how the viable system model (VSM) and de Sitter's design theory can complement each other in the context of the diagnosis and design of viable organizations. - Design/methodology/approach – Key concepts from Beer's model and de Sitter's design

  5. Analysis of a Polystyrene-Toluene System through "Dynamic" Sorption Tests: Glass Transitions and Retrograde Vitrification.

    Science.gov (United States)

    Pierleoni, Davide; Minelli, Matteo; Scherillo, Giuseppe; Mensitieri, Giuseppe; Loianno, Valerio; Bonavolontà, Francesco; Doghieri, Ferruccio

    2017-10-26

    Exposing a glassy polymer to a fluid phase (in gaseous or liquid state) containing a low molecular weight compound results in the sorption of the latter within the polymer, inducing, among other effects, the plasticization of the material which also promotes a change in the glass transition temperature. The amount of sorbed penetrant is often related in a complex fashion to the temperature and pressure of the fluid, thus determining that the locus of glass transition, when represented in pressure-temperature coordinates, may display as well rather complex patterns. This is an issue of particular importance in several applications of glassy polymers. In particular, we investigated the behavior of polystyrene in contact with toluene vapor by performing several modes of dynamic sorption experiments, in which the rate of change of the temperature of the system and/or of the pressure of the vapor phase are controlled with high accuracy, with the aim of creating a map of rubbery and glassy states of the polymer as a function of temperature and pressure of the toluene vapor. Isothermal tests were performed by changing the pressure at a controlled rate, isobaric tests were performed by changing the temperature at a controlled rate, and isoactivity tests were performed by concurrently changing, in a proper way, both temperature and pressure. A relevant feature resulting from these experiments is the presence of a discontinuity in the slope of the mass of toluene sorbed within polystyrene reported as a function of temperature and/or pressure. This discontinuity has been interpreted as the indication of the occurrence of a glass transition. The elaboration of the experimental results allowed identification of the pressure/temperature conditions at which rubbery or glassy states of the polymer mixture are established. Quite interestingly, the system displays the so- called "retrograde vitrification" phenomenon, which consists of the occurrence of a rubbery-to-glassy state

  6. Hymenolepis nana: immunity against oncosphere challenge in mice previously given viable or non-viable oncospheres of H. nana, H. diminuta, H. microstoma and Taenia taeniaeformis.

    Science.gov (United States)

    Ito, A; Onitake, K; Sasaki, J; Takami, T

    1991-04-01

    When mice, previously given oral inoculation with viable oncospheres of the heterologous cestode species (Hymenolepis diminuta, H. microstoma, Taenia taeniaeformis) and the homologous one (H. nana), were challenged with oncospheres of H. nana 4 days after the primary inoculation, they showed strong and complete resistance to H. nana challenge, respectively. However, the resistance was not evoked in mice given either infective eggs of Toxocara canis or non-viable oncospheres of all cestode species examined. Congenitally athymic nude mice given viable oncospheres did not show any resistance to H. nana either. Eosinophil infiltration around cysticercoids of H. nana in the intestinal villi appeared to be more prominent in mice previously given viable oncospheres of H. diminuta than in mice given non-viable oncospheres or PBS only. Some of the eosinophils in the villus harboring cysticercoid(s) of H. nana invaded the epithelia in the former, whereas all eosinophils remained in the lamina propria in the latter. There was almost no eosinophil infiltration in nude mice. Microscopic observations revealed that oncospheres of H. diminuta, which require beetles as the intermediate host like H. microstoma, could invade the mouse intestinal tissue. Therefore, it is strongly suggested that the strong cross resistance to H. nana in mice, induced by oncospheres of all heterologous cestode species, is thymus-dependent and due to oncospheral invasion into the intestinal tissue of mice.

  7. Technical Exchange on Improved Design and Performance of High Level Waste Melters - Final Report

    Energy Technology Data Exchange (ETDEWEB)

    SK Sundaram; ML Elliott; D Bickford

    1999-11-19

    SIA Radon is responsible for management of low- and intermediate-level radioactive waste (LILW) produced in Central Russia. In cooperation with Minatom organizations Radon carries out R and D programs on treatment of simulated high level waste (HLW) as well. Radon scientists deal with a study of materials for LILW, HLW, and Nuclear Power Plants (NPP) wastes immobilization, and development and testing of processes and technologies for waste treatment and disposal. Radon is mostly experienced in LILW vitrification. This experience can be carried over to HLW vitrification especially in field of melting systems. The melter chosen as a basic unit for the vitrification plant is a cold crucible. Later on Radon experience in LILW vitrification as well as our results on simulated HLW vitrification are briefly described.

  8. Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification.

    Science.gov (United States)

    Morató, Roser; Castillo-Martín, Míriam; Yeste, Marc; Bonet, Sergi

    2016-06-01

    The aim of our study was to assess whether the cryotolerance of in vitro-produced embryos could be influenced by the length of in vitro culture and size of blastocoel cavity before vitrification, using the pig as a model. For this purpose we analysed the cryoresistance and apoptosis rate of blastocysts at different stages of development as derived on Day 5 and 6 of in vitro culture. Blastocysts were subsequently vitrified, warmed and cultured for 24h. Re-expansion rates were recorded at 3 and 24h and total cell number and apoptotic cells were determined at 24h. Day-6 blastocysts showed the highest rates of survival after warming, which indicates higher quality compared with Day-5 blastocysts. Higher re-expansion rates were observed for expanded blastocysts and those in the process of hatching when compared with early blastocysts. Total cell number and apoptotic cells were affected by blastocyst stage, vitrification-warming procedures and length of in vitro culture, as expanding and hatching-hatched blastocysts from Day 6 presented higher percentages of apoptotic cells than fresh blastocysts and blastocysts vitrified at Day 5. Our findings suggest that the cryotop vitrification method is useful for the cryopreservation of porcine blastocysts presenting a high degree of expansion, particularly when vitrification is performed after 6 days of in vitro culture. Furthermore, these results show that faster embryo development underlies higher blastocyst cryotolerance and provide evidence that blastocoel cavity expansion before vitrification is a reliable index of in vitro-produced embryo quality and developmental potential.

  9. Application of evolved gas analysis to cold-cap reactions of melter feeds for nuclear waste vitrification

    Energy Technology Data Exchange (ETDEWEB)

    Kruger, Albert A.; Chun, Jaehun; Hrma, Pavel R.; Rodriguez, Carmen P.; Schweiger, Michael J.

    2014-04-30

    In the vitrification of nuclear wastes, the melter feed (a mixture of nuclear waste and glass-forming and modifying additives) experiences multiple gas-evolving reactions in an electrical glass-melting furnace. We employed the thermogravimetry-gas chromatography-mass spectrometry (TGA-GC-MS) combination to perform evolved gas analysis (EGA). Apart from identifying the gases evolved, we performed quantitative analysis relating the weighed sum of intensities of individual gases linearly proportional with the differential themogravimetry. The proportionality coefficients were obtained by three methods based on the stoichiometry, least squares, and calibration. The linearity was shown to be a good first-order approximation, in spite of the complicated overlapping reactions.

  10. Cryotop vitrification for in vitro produced bovine and buffalo (Bubalus bubalis embryos at different stages of development

    Directory of Open Access Journals (Sweden)

    B. Gasparrini

    2010-02-01

    Full Text Available The aim of this study was to evaluate the possibility to vitrify in vitro produced (IVP buffalo and bovine embryos at different stages of development by an advanced version of the “minimal volume approaches”: the Cryotop method. In both experiments, the embryos were vitrified at the tight morula (TM, early blastocyst (eBl, blastocyst (Bl, expanded blastocyst (xBl and, only for buffalo, at the hatched blastocyst (hBl stage. After warming, the embryos were cultured in vitro for 24 hours. Stage of development affected the freezability of IVP embryos of both species with the highest embryo survival rates at advanced stages (xBl=76% and hBl=75% for buffalos and xBl=75% for bovine. These results suggest that Cryotop vitrification is an efficient method for buffalo and bovine IVP embryo cryopreservation.

  11. Development of pyrometallurgical partitioning technology for TRU in high level radioactive wastes. Vitrification process for salt wastes

    Energy Technology Data Exchange (ETDEWEB)

    Sakamura, Yoshiharu; Inoue, Tadashi [Central Research Inst. of Electric Power Industry, Komae, Tokyo (Japan). Komae Research Lab.; Shimizu, Takafumi; Kobayashi, Kuniaki

    1997-12-31

    A vitrification process for chloride wastes generated in the pyrometallurgical partitioning of TRUs from high level radioactive wastes is being developed. In the process, chlorides are reduced to metals by molten salt electrolysis. The metals are oxidized by air and then vitrified. Lithium metal and chlorine gas are recycled. The behaviors of lithium, sodium and fission products during molten salt electrolysis were studied by using various compositions of salts and cathode materials. It was shown that every metal can be recovered into a liquid lead cathode, and that a liquid cadmium cathode and a solid cathode are suitable for recovering lithium and sodium metal, respectively. Based on the experimental results the process flow sheet was discussed. (author)

  12. Non-viable antagonist cells are associated with reduced biocontrol performance by viable cells of the yeast Papiliotrema flavescens against Fusarium head blight of wheat.

    Science.gov (United States)

    Microbially-based plant disease control products have achieved commercial market success, but the efficacy of such biocontrol products is sometimes deemed inconsistent. Improper processing of harvested microbial biomass or long-term storage can reduce the proportion of viable cells and necessitate t...

  13. Effects of Different Maturation Systems on Bovine Oocyte Quality, Plasma Membrane Phospholipid Composition and Resistance to Vitrification and Warming.

    Science.gov (United States)

    Sprícigo, José F W; Diógenes, Mateus N; Leme, Ligiane O; Guimarães, Ana L; Muterlle, Carolle V; Silva, Bianca Damiani Marques; Solà-Oriol, David; Pivato, Ivo; Silva, Luciano Paulino; Dode, Margot A N

    2015-01-01

    The objective of this study was to evaluate the effects of different maturation systems on oocyte resistance after vitrification and on the phospholipid profile of the oocyte plasma membrane (PM). Four different maturation systems were tested: 1) in vitro maturation using immature oocytes aspirated from slaughterhouse ovaries (CONT; n = 136); 2) in vitro maturation using immature oocytes obtained by ovum pick-up (OPU) from unstimulated heifers (IMA; n = 433); 3) in vitro maturation using immature oocytes obtained by OPU from stimulated heifers (FSH; n = 444); and 4) in vivo maturation using oocytes obtained from heifers stimulated 24 hours prior by an injection of GnRH (MII; n = 658). A sample of matured oocytes from each fresh group was analyzed by matrix associated laser desorption-ionization (MALDI-TOF) to determine their PM composition. Then, half of the matured oocytes from each group were vitrified/warmed (CONT VIT, IMA VIT, FSH VIT and MII VIT), while the other half were used as fresh controls. Afterwards, the eight groups underwent IVF and IVC, and blastocyst development was assessed at D2, D7 and D8. A chi-square test was used to compare embryo development between the groups. Corresponding phospholipid ion intensity was expressed in arbitrary units, and following principal components analyses (PCA) the data were distributed on a 3D graph. Oocytes obtained from superstimulated animals showed a greater rate of developmental (Pvitrification because the blastocyst rate at D7 was similar (P>0.05) for all groups (CONT VIT = 2.8±3.5%, IMA VIT = 2.9±4.0%, FSH VIT = 4.3±7.2% and MII VIT = 3.6±7.2%). MALDI-TOF revealed that oocytes from all maturation groups had similar phospholipid contents, except for 760.6 ([PC (34:1) + H]+), which was more highly expressed in MII compared to FSH (Pbovine oocytes to vitrification.

  14. The use of antifreeze protein type III for vitrification of in vitro matured bovine oocytes.

    Science.gov (United States)

    Chaves, Dowglish F; Campelo, Iana S; Silva, Mirelly M A S; Bhat, Maajid H; Teixeira, Darcio I A; Melo, Luciana M; Souza-Fabjan, Joanna M G; Mermillod, Pascal; Freitas, Vicente J F

    2016-12-01

    The aim of this study was to evaluate the use of antifreeze protein type III (AFP III) into vitrification medium on meiotic spindle morphology of in vitro matured bovine oocytes as well as the fertilization and blastocyst rates. Mature cumulus-oocyte complexes (COC) were distributed in four groups: control (untreated), vitrified without supplementation (AFP0) or supplemented with 500 (AFP500) or 1000 ng/mL (AFP1000) into vitrification solutions. Samples from each group were used to analyze the organization of meiotic spindle by confocal microscopy and the remaining COC were submitted to in vitro fertilization and culture for eight days. Control group exhibited only 15% of abnormal spindle. However, the spindle morphology was affected in all vitrified groups regardless to AFP concentration: 75.8%, 76.1% and 69.2% (P > 0.05) for AFP0, AFP500 and AFP1000, respectively. Similar cleavage rate was obtained among the vitrified groups (AFP0 = 17.9%, AFP500 = 16.9% and AFP1000 = 17.8%), but lower (P < 0.05) compared with control group (68.7%). At Day 5 of culture, embryo production rate of AFP500 (30.8%) and AFP1000 (25.0%) were similar to control group (49.4%). However, at Day 8 of culture, AFP0, AFP500 and AFP1000 groups exhibited lower (P < 0.05) blastocyst rates (10.0%, 3.8% and 9.4%, respectively) when compared to control (41.1%). In conclusion, AFP III did not preserve meiotic spindle organization against the cryoinjuries. However, the use of AFP III improved embryo development at Day 5 of culture, although this effect was not maintained up to the blastocyst formation. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Development of a method of vitrification, thawing, and transfer of mammalian blastocysts using a single closed cryo-straw.

    Science.gov (United States)

    Heo, Young Tae; Lim, Joon Kyo; Xu, Yong Nan; in Jang, Woo; Jeon, Soon Hong; Kim, Nam-Hyung

    2014-01-01

    There are different methods for cryopreservation of mammalian embryos with variable degrees of success. These methods require specific vessels for embryo vitrification, thawing, and transfer. Here, we report a simple and inexpensive way to vitrify, thaw and transfer mammalian blastocysts in one straw. This in-straw vitrification solution with microdrop (ISVDM) was compared with EM grid and normal 0.25 mL straw methods. There were no differences in the rates of re-expanded and hatching-to-hatched murine and bovine blastocysts exposed to 1, 0.5, and 0.3 M of sucrose in the diluent that was loaded into the straw. Low re-expanded and hatching-to-hatched rates of murine and bovine blastocysts were observed with PBS only. The pregnancy rates of control murine blastocysts (57.1%) and blastocysts exposed to 0.3 M sucrose in diluent and ISVDM (71.4%) were significantly higher (p < 0.05) than those exposed to 1, 0.5, and 0 M sucrose and those loaded into 0.25 mL straws. The rate of offspring delivery was highest in the control group. There was no significant difference (p < 0.05) in the rate of offspring delivery among ISVDM, 0.25 mL straw, and EM grid groups. Our results indicate that vitrified embryos can be warmed and diluted in a single straw and that this one-step method enables farm animal embryo transfer without a microscope or other laboratory equipment.

  16. Cryosurvival and pregnancy rates after exposure of IVF-derived Bos indicus embryos to forskolin before vitrification.

    Science.gov (United States)

    Sanches, B V; Marinho, L S R; Filho, B D O; Pontes, J H F; Basso, A C; Meirinhos, M L G; Silva-Santos, K C; Ferreira, C R; Seneda, M M

    2013-09-01

    In vitro-produced (IVP) bovine embryos are more sensitive to cryopreservation than their in vivo counterparts due to their higher lipid concentrations, whereas Bos indicus IVP embryos are even more sensitive than Bos taurus IVP embryos. To examine the effects of a lipolytic agent, before vitrification of Bos indicus IVP embryos, on embryo survival, viability, and pregnancy rates, two experiments were conducted. In experiment 1, Bos indicus (Nelore) embryos were produced from abattoir-derived ovaries and allocated into two groups. In the treatment group, 10 μM of forskolin was added to the in vitro culture medium on Day 5 and incubated for 48 hours. On Day 7 of culture, IVP-expanded blastocysts from both the control (n = 101) and treatment (n = 112) groups were vitrified with ethylene glycol and DMSO via the Cryotop procedure. Although there was no significant difference between the rates of blastocoel reexpansion and hatching of the embryos exposed to forskolin (87.5% and 70.5%, respectively) compared with the control embryos (79.2% and 63.3%, respectively), the numerically superior rates of the embryos exposed to forskolin led to another experiment. In experiment 2, blastocysts produced from the ovum pick up were exposed or not exposed to the lipolytic agent and vitrified as in experiment 1. Embryos treated with forskolin had higher pregnancy rates than the control group (48.8% vs. 18.5%). In view of these results, 1908 Bos indicus embryos were produced from ovum pick up, exposed to the lipolytic agent, and blastocysts were transferred to recipients, and the pregnancy rates of the embryos of various breeds were compared. The mean pregnancy rate obtained was 43.2%. All data were analyzed by chi-square or by binary logistic regression (P ≤ 0.05). In conclusion, treatment with forskolin before vitrification improved cryotolerance of Bos indicus IVP embryos, resulting in good post-transfer pregnancy rates. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. FINAL REPORT REGULATORY OFF GAS EMISSIONS TESTING ON THE DM1200 MELTER SYSTEM USING HLW AND LAW SIMULANTS VSL-05R5830-1 REV 0 10/31/05

    Energy Technology Data Exchange (ETDEWEB)

    KRUGER AA; MATLACK KS; GONG W; BARDAKCI T; D' ANGELO NA; BRANDYS M; KOT WK; PEGG IL

    2011-12-29

    The operational requirements for the River Protection Project - Waste Treatment Plant (RPP-WTP) Low Activity Waste (LAW) and High Level Waste (HLW) melter systems, together with the feed constituents, impose a number of challenges to the off-gas treatment system. The system must be robust from the standpoints of operational reliability and minimization of maintenance. The system must effectively control and remove a wide range of solid particulate matter, acid mists and gases, and organic constituents (including those arising from products of incomplete combustion of sugar and organics in the feed) to concentration levels below those imposed by regulatory requirements. The baseline design for the RPP-WTP LAW primary off-gas system includes a submerged bed scrubber (SBS), a wet electrostatic precipitator (WESP), and a high efficiency particulate air (HEPA) filter. The secondary off-gas system includes a sulfur-impregnated activated carbon bed (AC-S), a thermal catalytic oxidizer (TCO), a single-stage selective catalytic reduction NOx treatment system (SCR), and a packed-bed caustic scrubber (PBS). The baseline design for the RPP-WTP HLW primary off-gas system includes an SBS, a WESP, a high efficiency mist eliminator (HEME), and a HEPA filter. The HLW secondary off-gas system includes a sulfur-impregnated activated carbon bed, a silver mordenite bed, a TCO, and a single-stage SCR. The one-third scale HLW DM1200 Pilot Melter installed at the Vitreous State Laboratory (VSL) was equipped with a prototypical off-gas train to meet the needs for testing and confirmation of the performance of the baseline off-gas system design. Various modifications have been made to the DM1200 system as the details of the WTP design have evolved, including the installation of a silver mordenite column and an AC-S column for testing on a slipstream of the off-gas flow; the installation of a full-flow AC-S bed for the present tests was completed prior to initiation of testing. The DM1200

  18. Immediate natural tooth pontic: A viable yet temporary prosthetic solution: A patient reported outcome

    Directory of Open Access Journals (Sweden)

    Sudhir Bhandari

    2012-01-01

    Conclusion: The concept of immediate pontic placement is surely a viable treatment option and promises an excellent transient esthetic solution for a lost tooth as well as enables good preparation of the extraction site for future prosthetic replacement.

  19. Improved identification of viable myocardium using second harmonic imaging during dobutamine stress echocardiography

    NARCIS (Netherlands)

    F. Sozzi (Fabiola); D. Poldermans (Don); J.J. Bax (Jeroen); A. Elhendy (Abdou); E.C. Vourvouri (Eleni); R. Valkema (Roelf); J. de Sutter; A.F.L. Schinkel (Arend); A. Borghetti; J.R.T.C. Roelandt (Jos)

    2001-01-01

    textabstractOBJECTIVE: To determine whether, compared with fundamental imaging, second harmonic imaging can improve the accuracy of dobutamine stress echocardiography for identifying viable myocardium, using nuclear imaging as a reference. PATIENTS: 30 patients with chronic left

  20. Marine environmental pollution stress detection through direct viable counts of bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah, N.; Kenkre, V.D.; Verlecar, X.N.

    Direct viable counts (DVC) of bacteria were quantified from polluted and relatively less/non-polluted coastal locations during different seasons to assess whether they can be routinely monitored for an understanding of environmental stress(es...