Abstract: Several physical stresses kill cells at low temperatures Intracellular ice is usually fatal, so survival of freezing temperatures involves combinations of dehydration, freezing point depression, supercooling and intracellular vitrification Artificial cryopreservation achieves intracellular vitrification with rapid cooling, modest osmotic contraction and, often, added cryoprotectants High warming rates are required to avoid crystallization during warming Environmental cooling is much slower and temperatures less cold, but environmental freezing damage is important ecologically and agronomically For modest sub-freezing temperatures, supercooling sometimes allows survival At lower temperatures, extracellular water usually freezes and cells may suffer large osmotic contractions This contraction concentrates solutes and thus assists vitrification, but is not necessarily reversible: the rapid osmotic expansion during thawing may rupture membranes Further, membranes and other ultrastructural elements may be damaged by the large, anisotropic mechanical stresses produced when their surfaces interact via hydration forces Solutes reduce these stresses by osmotic, volumetric and other effects

Abstract: This review contains two parts. The first part is devoted to the significant steps in cryopreservation of mammalian embryos with emphasis on cattle and sheep that serve as models of reference. These steps are: (1) shortening of cooling and warming processes; (2) addition and dilution of cryoprotectant in one step; (3) introduction of plastic straw as a freezing and dilution container; (4) the choice of ethylene glycol as the quite universal cryoprotectant because of its low toxicity and high permeability; (5) vitrification, a cryopreservation method which enable passage from the liquid to the solid state by extreme elevation of viscosity due to high concentration of cryoprotectants and very rapid cooling. There are several vitrification solutions which contain dimethyl sulphoxide, glycerol, ethylene glycol, or a mixture of them, as basic cryoprotectants. The second part considers some factors affecting the efficiency of cryopreservation concerning (i) the origin of embryos and (ii) the stage of development and species. The origin of embryos (in vivo versus in vitro): in vitro embryos show a chilling and freezing sensitivity associated with their lipid content which can be modified by the culture conditions. Both conventional freezing and vitrification have been used and it seems that vitrification is more adapted to in vitro embryos when some modifications of initial protocols are carried out, particularly the rate of cooling. Thus considerable progress has been achieved by using the open pulled straw method of Vajta which enables the use of a minimum volume of freezing medium (0.5 microl) and a very high cooling rate that permits rapid traversal of the damaging temperature zone, corresponding to chilling sensitivity. The stage of development and species: not only are there differences between species at the same stage of development but in the same species all stages of development do not survive equally under the same freezing protocol. In cattle for example, oocytes and early stages of development in vivo or in vitro do not survive whereas compacted morulae and blastocysts survive very well. In the pig hatched blastocysts survive better than the other stages. Horse embryos have special characteristics that pose problems for successful freezing. In conclusion, a lot of work remains to be done to define fundamental characteristics of embryos of certain species (pig, horse) and of embryos of some stages or of oocytes.

Abstract: Glass matrix composites have been obtained by mixing and sintering “negative cost” materials coming from municipal solid waste incinerators (MSWIs) and from one aluminium foundry. The bottom ashes from two MSWIs were used to obtain the glass matrix and the solid wastes from an aluminium alloy industry were employed as a second phase. The bottom ashes were vitrified by heating at 1400°C without any additive. The vitrification process reduces the bottom ash initial volume by about 60%. The obtained glass has a Young modulus of 96 GPa and a Vickers hardness of 6 GPa. The composites were prepared in air by a low temperature pressure-less viscous phase sintering process (740–830°C) and were morphologically and mechanically characterised. The composites showed a bending strength up to 50 MPa at room temperature and could be proposed for applications in the field of tiles.

Abstract: Increasing amounts of municipal solid waste incineration (MSWI) residues are treated prior to landfilling or reuse. In Japan, electric arc melting is used for bottom ash vitrification that generates a glasslike slag. The objective of this paper was to assess this pretreatment technique with respect to its effect on metal mobility and metal content. Both bottom ash and slag were sampled and analyzed on total solids (TS), fixed solids (FS), particle density (ρP), specific BET surface area, particle size distribution, and total element content. A six-step wet sequential extraction procedure was used for assessing metal mobility. The results were qualitatively verified by scanning electron microscopy. The major conclusion was that the availability of various metals was affected differently by electric arc vitrification. Metals were solidified, stabilized, and/or separated from the slag. The mobility of Cr, Cu, Zn, Pb, and Ca was reduced. In slag, major fractions of these elements were found in moderately redu...

Abstract: Recently, new vitrification methods for bovine embryos characterized by ultra-rapid cooling have been reported. The present study was undertaken to determine the availability of gel-loading tips as containers for vitrification of in vitro-produced bovine embryos at various developmental stages. Bovine embryos at Days-1, -2, -3, -4, -5, and -7 (the day of insemination was defined as Day-0) were placed in 0.6~0.7 μl of vitrification solution (20% ethylene glycol, 20% DMSO and 0.6 M sucrose) in a commercially available gel-loading tip, and cooled by immersing in liquid nitrogen. Survival of the vitrified embryos was evaluated by in vitro development into the blastocyst stage. There was no significant difference (P ≥0.05) between the survival rates for fresh and vitrified embryos at Day-1 (70.5 and 58.0%, respectively), Day-2 (75.9 and 68.3%, respectively), Day-3 (75.9 and 57.9%, respectively), Day-4 (80.0 and 67.6%, respectively), Day-5 (82.1 and 60.0%, respectively) and Day-7 (100 and 97.8%, respectively). And no significant differences were found between the total numbers of cells or the proportion of inner cell mass cells of blastocysts developed from fresh versus vitrified embryos. These results demonstrate that the gel-loading tip is applicable as a container for vitrification of in vitro-produced bovine embryos at the 2-cell to blastocyst stages.

Abstract: A method for vitrification of a tissue or organ other than a blood vessel includes immersing the tissue or organ in increasing concentrations of cryoprotectant solution at a temperature greater than - 15 °C to a cryoprotectant concentration sufficient for vitrification; cooling the tissue or organ at an average rate of from 2.5-100 °C per minute to a temperature between - 80 °C and the glass transition temperature; and further cooling the tissue or organ at an average rate less than 30 °C per minute to a temperature below the glass transition temperature to vitrify the tissue or organ. After the vitrified tissue or organ has been stored, the tissue or organ may be removed from vitrification by warming the tissue or organ at an average rate of from 20-40 °C per minute to a temperature between - 80 °C and the glass transition temperature; further warming the tissue or organ at a rate of from 200-300 °C per minute to a temperature above - 75 °C; and reducing the concentration of the cryoprotectant.

Abstract: Highly filled thermosets are used in applications such as integrated circuit (IC) packaging. However, a detailed understanding of the effects of the fillers on the macroscopic cure properties is limited by the complex cure of such systems. This work systematically quantifies the effects of filler content on the kinetics, gelation and vitrification of a model silica-filled epoxy/amine system in order to begin to understand the role of the filler in IC packaging cure. At high cure temperatures (100°C and above) there appears to be no effect of fillers on cure kinetics and gelation and vitrification times. However, a decrease in the gelation and vitrification times and increase the reaction rate is seen with increasing filler content at low cure temperatures (60-90°C). An explanation for these results is given in terms of catalysation of the epoxy amine reaction by hydrogen donor species present on the silica surface and interfacial effects.

Abstract: The sodium bearing tank waste (SBW) at Idaho National Engineering and Environmental Laboratory (INEEL) contains high concentrations of sulfur (roughly 5 mass% of SO3 on a nonvolatile oxide basis). The amount of sulfur that can be feed to the melter will ultimately determine the loading of SBW in glass produced by the baseline (low-temperature, joule-heated, liquid-fed, ceramic-lined) melter. The amount of sulfur which can be fed to the melter is determined by several major factors including: the tolerance of the melter for an immiscible salt layer accumulation, the solubility of sulfur in the glass melt, the fraction of sulfur removed to the off-gas, and the incorporation of sulfur into the glass up to it?s solubility limit. This report summarizes the current status of testing aimed at determining the impacts of key chemical and physical parameters on the partitioning of sulfur between the glass, a molten salt, and the off-gas.

Abstract: 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 Laboratory'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.

Abstract: The role of pregrowth and preculture treatments in terms of both medium composition and exposure duration on survival of embryonic axes of Citrus madurensis after cryopreservation using the vitrification procedure was investigated. The optimal pregrowth treatment for excised embryonic axes was a 3-day treatment with 0.1M sucrose. Preculture was also essential in increasing survival after cryopreservation. Among the various media and treatment durations evaluated, a 24h-preculture of embryonic axes on medium with 0.3M sucrose and 0.5M glycerol was found to be optimal. Using these pregrowth and preculture conditions followed by treatment at 25 degrees C for 20 min each with a loading solution (0.4M sucrose + 2.0M glycerol) and then the PVS2 vitrification solution, direct immersion in liquid nitrogen, rapid rewarming, unloading in a 1.2M sucrose solution for 20 min and transfer of embryonic axes on recovery medium, 82.5% survival and regrowth without intermediary callus formation were obtained with C. madurensis embryonic axes.

Abstract: A new method of cryopreservation based on the very fast cooling rates achieved by the direct contact of small droplets of vitrification solution containing biological sample with a very cold solid surface.

Abstract: The electrical behaviour of different glass compositions obtained from mixing waste glass deriving from a community glass recycling program and silicate waste from the incineration of municipal solid residues (from Reggio Emilia city) have been investigated as a function of temperature and frequency. The electrical and dielectric properties were related to structural studies performed on the same glassy materials. As the amount of incinerator wastes increases, on account of lower alkali and higher alkaline-earth content in the final glass composition, conductivity and dielectric losses decrease approaching the behaviour of type E glass fibres, so envisaging a possible use of waste-containing glasses in the production of high voltage insulators.

Abstract: The effects of fish antifreeze protein AFP-I on cryopreservation of rice suspension cells by three different protocols were investigated. During the two-step method, AFP-I at 0.01 mg/ml significantly lowered the viability of both precultured and non-precultured cells. During the vitrification method, AFP-I at 0.2 mg/ml improved the viability of suboptimally thawed cells; however, much higher doses of this protein (10mg/ml) attenuated the cell viability. During rapid freezing of rice cells in the solutions with relatively high (but non-vitrifying) concentrations of cryoprotectant, AFP-I displayed protective action in the higher concentrated cryoprotectants and detrimental effect in more dilute ones. Taken together, it was concluded that, depending upon a number of factors discussed in the present paper, both positive effect and negative effect could be observed during application of AFP to cryopreservation of rice cells. The possible mechanism of this dual character was discussed.

Abstract: The in vitro and in vivo development of mouse morulae after cryopreservation through different methods was examined. The slow-freezing involved an equilibration in 1.5M ethylene glycol (EG) and cooled at 0.5; 0.7; 1.0 or 1.2oC/minute. The vitrification involved a 3 minutes equilibration in 20% EG and 60 seconds in solution containing 40% EG, 18% ficoll and 10.26% sucrose. The quick-freezing involved an equilibration in 3M EG + 0.3M sucrose for 5 minutes and 2 minutes in nitrogen vapor. In all three methods the straws were thawed in air for 10 seconds and in water at 25oC for 20 seconds and the embryos cultured in vitro for 72 hours to estimate blastocyst rate. To assess viability in vivo, frozen morulae as well as fresh embryos were transferred into recipients. The in vitro development rates with 0.5, 0.7; 1.0 and 1.2oC/minute were, respectively, 72.3; 79.6; 76.5 and 84.8%. There was no significant difference among the cooling rates of 0.7; 1.0 and 1.2oC/minute (p > 0.01). The in vitro survival rates of vitrification and quick-freezing (84.5 and 74.3%, respectively) were similar to the slow-freezing. In vivo, the implantation rate and number of fetuses from embryos frozen through slow-freezing at 1.2oC/minute, vitrification and quick-freezing were not significantly different.

Abstract: Technologies are being developed by the US Department of Energy's (DOE) Nuclear Facility sites to convert low-level and mixed (hazardous and radioactive) wastes to a solid stabilized waste form for permanent disposal. One of the alternative technologies is vitrification into a borosilicate glass waste form. The Environmental Protection Agency (EPA) has declared vitrification the Best Demonstrated Available Technology (BDAT) for high-level radioactive mixed waste and produced a Handbook of Vitrification Technologies for Treatment of Hazardous and Radioactive Waste. The DOE Office of Technology Development (OTD) has taken the position that mixed waste needs to be stabilized to the highest level reasonably possible to ensure that the resulting waste forms will meet both current and future regulatory specifications. Stabilization of low level and hazardous wastes in glass are in accord with the 1988 Savannah River Technology Center (SRTC), then the Savannah River Laboratory (SRL), Professional Planning Committee (PPC) recommendation that high nitrate containing (low-level) wastes be incorporated into a low temperature glass (via a sol-gel technology). The investigation into this new technology was considered timely because of the potential for large waste volume reduction compared to solidification into cement.