Combination Of Ethylene Glycol Research Articles

Experimental Cooling Performance Evaluation of Different Coolants for Data Centre

The direct liquid cooling (DLC) of the data center is becoming popular due to its higher heat removal from the computer chips. The direct liquid cooling method is more effective than the conventional air-cooling system and reduces the higher infrastructure and maintenance costs. The DLC reduces the chip failure rate drastically and increases the life of the data centers. Different liquids can be used as a coolant and some manufacturers are coming up with different coolants where the liquid has high thermal efficiency and is electrically non-conductive. In this article, a heat-transfer cold plate made of aluminum is designed and a different combination of heat-transfer liquids (Distilled water, Ethylene Glycol, and Polyethylene Glycol) is tested to find a comparatively better combination of heat-transfer liquid. It was observed that the combination of Ethylene Glycol and distilled water performs better than other combinations. It was also found that the coolant flow rate plays an important role in the cooling of the chips as well.

Cryoprotective Effects of Longan Honey on Preantral Follicle Integrity of Rat Ovary Post Vitrification

BACKGROUND: Longan honey (LH) has the potential as a natural extracellular cryoprotectant to maintain the integrity of intact preantral follicles against the cryoinjury during ovarian vitrification. OBJECTIVE: This research determined the cryoprotective effects of logan honey on preantral follicles integrity of rat ovary post-vitrification. MATERIALS AND METHODS: After vitrification, the follicle index was determined by observing the ovarian histological sections made using the paraffin method with hematoxylin-eosin staining and analyzed using Optilab 3.0 and Image Raster software. RESULTS: The results showed that the combination of ethylene glycol (EG) with LH and a dose variation of 7.5%-15% (KP1-KP4) increased the density of follicles, the number of intact follicles in G2 and G3, with a decrease in the atretic follicles in G1 better compared to the use of EG only (KKP1-KKP2). The differences in the histological structur e of preantral follicles affected the doses of LH needed by each type of follicle to maintain the integrity of the follicular structure from cryoinjury effects. The comparison of the G2 total follicle index values were KKP1 (90.7±18.3), KKP2 (115.6±9.9) < KP1 (193.6±10.7), KP2 (189.3±10.5), KP3 (182.2±27.1) and KP4 (169.4±8.8). Among the variations in the dose of LH 7.5%-15%, the highest increase in the G3 index value was found in primary (51.653±9.791), tertiary (43.119±8.786), secondary (33.885±4.745), and primordial (28.670±2.516) follicles of KP3 (7.5% of LH). CONCLUSION: The primary and tertiary follicular stages maintain the best integrity and can be used after the vitrification of rat ovaries.

Discerning Comparison of 1 and 0.5% Ethylene Glycol in Sprague-Dawley Rats with Modeled Urolithiasis.

A common method of modeling urolithiasis is the use of 1 and 0.75% ethylene glycol, or a combination of ethylene glycol with other lithogens, but too rapid progression of the disease and multiple organ toxicity have been reported. We developed a urolithiasis model in Sprague-Dawley rats, in which the animals received a relatively low concentration of ethylene glycol (0.5%), but for a long-term period (6 weeks) followed by animal observation during the 6-week recovery period. In urine samples, signs of the urolithiasis development were observed starting from the sixth week: the presence of ketones, decrease in diuresis and urine pH; in the blood, urea, protein, and hematocrit were elevated. However, no leukocytes were detected in the urine; in the blood, no shifts in differential leukocyte count and no elevation in ALT, creatinine, cholesterol, and triglycerides were observed, which indicates the absence of multiple organ failure while using 1% ethylene glycol. In addition, the animals receiving 0.5% ethylene glycol were followed up to 12 weeks in contrast to animals receiving 1% ethylene glycol (the experiment in this case was stopped during the third week for ethical reasons).

P-220 Ice crystals propagation velocity is related to the type and concentration of cryoprotectants and impacts cells and tissue vitrification success

Abstract Study question How different cryoprotectants at various concentrations affect ice crystals formation and propagation during vitrification? Summary answer The velocity of ice crystal propagation is inversely related to concentration and type of cryoprotectants and is responsible for downstream cell damage during vitrification. What is known already Vitrification has replaced slow freezing for oocyte and embryo cryopreservation and is now being considered also for gonadal tissue. However, the results in terms of both vitrification and rewarming survival rates are still not uniform. Understanding the complex physical events controlling ice crystals formation, propagation and expansion are key for improving cryoprotectant solutions and overcoming challenges. Study design, size, duration Basic science research. Ice crystal propagation velocity was measured in solutions of four cryoprotectants: dimethyl sulfoxide (DMSO), propylene glycol (PG), ethylene glycol (EG) and glycerol (Gly), at various concentrations in supercooled temperatures. Each experiment used five 0.25ml and five 2ml straws (volume 0.1 ml and 1 ml) containing different cryoprotectant combinations (DMSO:EG, EG:PG, DSMO:PG, DMSO:Gly, EG:Gly, PG:Gly and EG+PG vs EG+DMSO), immersed in alcohol baths prepared at 8-10 °C below the freezing points of each solution. Participants/materials, setting, methods Alcohol baths were set as follows: (1) between -10 °C and -12 °C for straws with 0% and 10% (v/v) DMSO, EG, PG and Gly; (2) -15 °C to -19 °C for the 20%; (3) -19.5 to -24 °C for the 30%, and for the mixture of 15% EG and 15% PG and 15% EG and 15% DMSO (most used vitrification solutions); and (4) -25 to -36 °C for the 40%. A 50µm thermocouple measured supercooling. Student’s T-test analyzed differences. Main results and the role of chance Seeding was done by touching the straw end with LN- precooled forceps until visualizing a small ice crystal. Following 10 to 60s the distance to which the ice crystals grew inside the straws was evaluated using a fine caliber. Ice crystal propagation was inversely correlated to type and cryoprotectant concentrations. Propylene glycol showed statistically significant lower ice crystals growth velocity up to concentrations of 30% (v/v), compared to DMSO and EG at the same concentrations (p < 0.05). Moreover, the ice crystal velocity of PG was also lower than Gly at these same concentrations. The combination of EG with PG decreased ice crystal propagation better than EG with DMSO. At concentrations of 40%, there were no differences. These results suggest that current cryopreservation vitrification solutions are not optimized and combining PG with other CPs has advantages. Using a vitrification solution of 40% DMSO or EG, the velocity of crystal growth was in the range of 0.1mm/sec (6mm/min). This means that to vitrify large tissues, a cooling rate of over 1000 °C/min from RT to a temperature of -196 °C (ΔT of 220 °C in 13 seconds) is necessary to avoid a single ice nucleus to grow to a size of 1.3mm during cooling or warming. Limitations, reasons for caution Assessing the velocity of ice propagation is a novel concept to perfect vitrification/warming solutions. Wider implications of the findings These results pave the way to develop protocols and platforms able to minimize the deleterious impact of cryoprotectant concentrations on both cells and tissues survival after vitrification. Trial registration number not applicable

Experimental and Numerical studies on the Heat transfer characteristics of Small Shell and Tube Heat Exchanger

This work presents the experimental and numerical investigations on the effectiveness of a small shell and tube heat exchanger for different coolant combinations. A numerical investigation is done using commercial computational tool, Fluent, to analyze heat transfer characteristics for the selected coolant combinations. The coolant combinations used consists of ethylene glycol (EG) and deionized water (DW) with varied percentages of the former. Accordingly, combination of ethylene glycol (EG) and deionized water viz, 60EG:40DW, 30EG:70DW and 20EG:80DW were used. The base fluids of EG and DW were used for the comparison of heat transfer characteristics, respectively with the selected coolant combination. For the numerical investigations, the CAD model of counter flow heat exchanger with 0 degree inclined baffles; with cold liquid flowing through the shell at the rate of 0.3 kg/s while hot liquid flows through the copper tubes at a flow rate of 0.3 kg/s was considered. Different combinations of shell-tube liquid are analyzed for identical initial and boundary conditions, keeping all other Fluent parameters the same. The temperature contours are generated for all the five cases. The temperature distribution for different shell-tube liquid combinations are compared for evaluating the effectiveness of heat exchange.Improved heat exchange is ensured with higher temperature difference between inlet and outlet of shell/tube, and respective combination of shell and tube fluid is selected for industrial cooling applications. Results of CFD approach are validated with the experimental results and results are acceptable. From the results it is found that 60EG:40DW combination showed improved heat transfer characteristics compared to other combinations used. Among the different coolant combinations, 60EG:40DW showed higher heat transfer as indicated by higher temperature difference across both shell and tube sections followed by other combinations when compared to pure ethylene glycol. Shell outlet and Tube outlet temperature with 60EG:40DW increased by 227% and 105% respectively when compared with pure EG, due to improved heat exchange. Heat transfer rate, heat transfer coefficient with 60EG:40DW increased when compared to pure EG.

An insight into the performance of radiator system using ethylene glycol-water based graphene oxide nanofluids

Recent studies showed that nanofluids are considered as a promising way to enhance the heat transfer properties of coolants. Currently, in a car radiator forced convection heat transfer method is carried out to cool circulating fluid. The mixture of water with either ethylene or propylene-glycol is necessary to lower its freezing point and eliminate ice formation. However, the boiling point of deionized water can be pushed up by mixing it with anti-freezing materials like ethylene glycol (EG). Hence the present research work focuses on the heat transfer characteristics of nano-fluids with different combinations of EG and deionized water (60:40, 30:70, and 20:80) along with graphene oxide (GO) nanoparticles (0.1 % wt). The heat transfer potential of graphene oxide/ deionized water-ethylene glycol nanofluids is experimentally investigated in coolant application for car radiators, in which the coolant flow rate is varied from 180 to 420 (LPH) at a fixed coolant temperature of 90 °C. The evaluation of stability is performed using visual inspection and zeta potential test. The density of nanofluid was determined by oscillating U-Tube density meter. The optimized combination of 60% EG, 40% DW and 0.1 wt% GO exhibited higher heat transfer characteristics of the radiator system used. The maximum heat transfer enhancement of 42.77% at 300 LPH, 18.14% at 360 LPH, and 71.1% at 240 LPH for 60:40 EG/DW-based GO nanofluids were obtained. The maximum Nusselt number value of 192 at 420 LPH was observed for 60:40 EG/DW-based GO nanofluid as compared with 20:80, 30:70 EG/DW-based GO nanofluid. It is estimated that by replacing conventional coolant with this nanofluid, reduction in the frontal area of radiator is done which gives more flexibility for industrial design, more eco-friendly vehicle which produces less drag and hence less the fuel cost.

Enhanced Power Factor of PEDOT:PSS Films Post-treated Using a Combination of Ethylene Glycol and Metal Chlorides and Temperature Dependence of Electronic Transport (325–450 K)

We report temperature dependence of electrical conductivity and thermopower for poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) thin films post-treated using a combination of an...

Vitrification of collared peccary ovarian tissue using open or closed systems and different intracellular cryoprotectants

This study aimed to evaluate different vitrification methods using distinct cryoprotectants (CPAs) for the preservation of collared peccary ovarian preantral follicles (PFs). Ovarian pairs from six females were fragmented and three fragments (fresh control group) were immediately evaluated for morphology, viability, cell proliferation capacity (assessed by quantifying the number of argyrophilic nucleolus organizer regions – NORs), and apoptosis (by the identification of activated caspase-3 expression). The remaining 18 fragments were vitrified using the solid surface vitrification (SSV) method or the ovarian tissue cryosystem (OTC) with 3 M ethylene glycol (EG), 3 M dimethylsulfoxide (DMSO), or a combination of the two (1.5 M EG/1.5 M DMSO). After two weeks, samples were rewarmed and evaluated as described previously. The OTC with any of the CPAs provided a similar conservation of morphologically normal PFs as the fresh control group (75.6 ± 8.6%); however, the SSV was only efficient with DMSO alone (63.9 ± 7.6%). Regarding the viability or cell proliferation, all tested groups provided post rewarming values similar to those observed for the fresh control group, 84.0 ± 2.9% viable cells with 2.0 ± 0.2 NORs. Related to apoptosis analysis, only the OTC with EG (46.7%) and the SSV method with EG (43.4%) or the combination of EG and DMSO (33.4%) provided similar values to those found for the fresh control group (36.7%). Our findings indicate the utilization of a closed system, the OTC, with 3 M EG as the CPA for the vitrification of collared peccary ovarian tissue.

Generation of Micro/Nano Scale Particles and Capsules by Electro hydrodynamic Atomization (EHDA) Process

Abstract Electro hydrodynamic atomization (EHDA) process is a technique of generating liquid droplets through the application of a large electrical potential difference. It has a wide range of industrial, agricultural and medical applications. This paper deals with the application of EHDA for generation of particles and capsules in micro/nano scale. Flow visualization is conducted to report different modes appearing during the process. The flow is visualized for different operating conditions, both above and below the grounded ring electrode. It is observed that the number of modes of e-spray increases with decreasing electrical conductivity of fluid. Micro particles are generated for different set of fluids. The sizes of generated particles increase nonlinearly with increasing flow rate for a fluid. It is investigated that the charge carried by droplets in DC elecrospray increases/decreases sharply when the mode of e-spray changes, unlike AC electrospray where generated droplets are neutral in nature. The capsules are generated for the combination of ethylene glycol, PCL solution with sodium silicate solution.

Sifat Serbuk dan Sifat Elektrik Bahan Katod La0.6Sr0.4CoO3-δ yang Disediakan Melalui Kaedah Sol-Gel Terubahsuai Bagi Aplikasi Sel Fuel Oksida Pepejal

The powder properties and electrical conductivity of lanthanum strontium cobalt oxide, La0.6Sr0.4CoO3-δ (LSC) material were systematically characterized with respect to application as cathode material for intermediate temperature solid oxide fuel cell (SOFC). The LSC was prepared via a modified sol-gel method assisted with a combination of ethylene glycol and activated carbon as a chemical additive. The decomposition of impurity compounds such carbon compounds and nitrates in the precursor powder was completed at 870 °C as revealed by thermogravimetry (TG) analysis. As the calcination temperature increased from 800 °C to 1000 °C, the purity and crystallite size of the the calcined precursor powder also increased as confirmed by X-ray diffrcation (XDR) analysis. The micrograph of scanning electron microscope (SEM) image showed that the particle of the calcined powders form agglomeration and this led to low relative density (80.18 %) of the sintered LSC pellet at 1200 °C. The direct current electrical conductivities (σdc) of the sintered LSC pellet measured using Van der Pauw technique in air were 1504 S cm-1 and 1069 S cm-1 at 400 °C and 800 °C, respectively. The relatively high σdc with low activation energy (Ea) of 0.021 eV indicates that the LSC prepared in this work has a great potential to be used as a cathode material for intermediate temperature SOFC application.

Combination of ethylene glycol with sucrose increases survival rate after vitrification of somatic tissue of collared peccaries (Pecari tajacu Linnaeus, 1758)

ABSTRACT: The cryopreservation of somatic tissue in collared peccaries promotes an alternative source of genetic material of this specie. The solid-surface vitrification (SSV) is a great option for tissue conservation; nevertheless, the optimization of SSV requirements is necessary, especially when referred to cryoprotectants that will compose the vitrification solution. Therefore, the aim was to evaluate the effect of the presence of 0.25 M sucrose in addition to different combinations (only or association) and concentrations (1.5 M or 3.0 M) of ethylene glycol (EG) and/or dimethyl sulfoxide (DMSO) in the somatic tissue vitrification of collared peccaries. Subsequently, we tested six combinations of cryoprotectants with or without sucrose in Dulbecco modified Eagle medium (DMEM) plus 10% fetal bovine serum (FBS). Thus, 3.0 M EG with sucrose was able to maintain normal tissue characteristics compared with non-vitrified (control), especially for the volumetric ratio of epidermis (61.2 vs. 58.7%) and dermis (34.5 vs. 36.6%), number of fibroblast (90.3 vs. 127.0), argyrophilic nucleolar organizer region (AgNOR) ratio (0.09 vs. 0.17%) and nucleus area (15.4 vs. 14.5 μm2) respectively. In conclusion, 3.0 M EG with 0.25 M sucrose and 10% FBS resulted in a better cryoprotectant composition in the SSV for somatic tissue of collared peccaries.

Effects of highly conductive PH1000 anode in combination with ethylene glycol additive and H2SO4 immersion treatments on photovoltaic performance and photostability of polymer solar cells

In this study, we have fabricated efficient polymer solar cells (PSCs) by introducing a highly conductive poly(3,4-ethylene dioxy-thiophene):poly(styrene sulfonate) (PH1000) thin film treated with a combination of ethylene glycol (EG) additive and H2SO4 solution immersion as a transparent electrode (PH1000–EG–H2SO4).

48 Effect of Dimethyl Sulfoxide- or Glycerol-Based Vitrification Protocols on Meiotic Spindle of In Vitro-Matured Bovine Oocytes

There is evidence suggesting that high concentrations of cryoprotectants (CPA) and very low temperatures during vitrification cause disruption of the meiotic spindle, resulting in poor post-warming meiotic resumption and other abnormalities at fertilization. This study sought to determine the damage caused by CPA and freezing upon the meiotic spindle of bovine oocytes vitrified at the metaphase II stage, and whether a subsequent incubation could promote recovery from this damage. Bovine cumulus–oocyte complexes were purchased from a commercial vendor (n = 154). Oocytes were removed from in vitro maturation media at 22 h, denuded by vortexing in hyaluronidase, and divided into 4 groups according to CPA exposure and whether they were incubated or not. The resulting groups were DMSO I (n = 36), DMSO NI (n = 41), GLY I (n = 39), GLY NI (n = 38). Two repetitions were carried out for each protocol evaluated, which included a combination of ethylene glycol (EG) with either dimethyl sulfoxide (DMSO) or glycerol (GLY). Oocytes were exposed to equilibration solution consisting of 7.5% EG and 7.5% DMSO or GLY for 9 min at room temperature (RT) and then placed into vitrification solution (VS) that contained 15% EG, 15% DMSO or GLY, and 0.5 M sucrose. While in VS, 3 to 4 denuded oocytes were loaded onto a Cryolock® (Biotech Inc., Alpharetta, GA, USA) and plunged into liquid nitrogen within 1 min. For warming, oocytes were exposed to previously warmed (37°C) dilution solution 1 (DS1) consisting of 0.5 M sucrose for 1 and 2 min at RT, for a total of 3 min in DS1, and then placed in dilution solution 2 containing 0.25 M sucrose for 3 min. Finally, oocytes were washed in base media. Base media for all solutions was PBS supplemented with 20% fetal bovine serum. After warming, half of the oocytes were fixed and the rest were submitted to a 2-h incubation period in maturation media at 37°C and 5.5% CO2, and then fixed. To examine microtubule distribution and chromosome arrangement, fixed oocytes were submitted to an immunofluorescence protocol using α-tubulin antibody (1:100) as primary antibody, Alexa Fluor 488 (1:1000) as secondary antibody, and counterstained with propidium iodide (10 mg mL−1). Oocytes were observed under a fluorescence microscope. The effects of CPA and incubation on the incidence of abnormal spindles measured with both microtubules distribution and chromosome arrangement were evaluated using logistic regression with a binomial response variable (normal/abnormal). For microtubule distribution, results showed that oocytes treated with DMSO presented significantly lower normality (31.17%) than those treated with glycerol (54.55%; P < 0.003). The most common abnormality observed in oocytes treated with DMSO was that the spindle was smaller and more faintly stained than those treated with glycerol. For chromosome arrangement, there was no significant difference between treatments (P = 0.7093). Additionally, there was no sign of improvement when submitting the oocytes to an incubation period for any of the components examined.

Effect of cryoprotectants and thawing temperatures on chicken sperm quality.

There is need for standardization of freezing-thawing protocol for rooster semen to minimize variability among results. Therefore, we aimed to compare effect of four different permeating cryoprotectants and two thawing temperatures (37 vs. 5°C) on sperm post-thaw motility and to analyse combined effect of the best permeating cryoprotectant (P-CPA) with one of four non-permeating cryoprotectants (N-CPA) on post-thaw quality of rooster semen evaluated in vitro. Pooled semen from Ross PM3 rooster heavy line was diluted in Kobidil extender and frozen in cryoprotectant solution containing 6% dimethylacetamide, 7.5% dimethylformamide, 9% N-methylacetamide or 8% ethylene glycol (EG) in liquid nitrogen vapours. To determine the best thawing rate, straws were thawed either at 37 or 5°C. Furthermore, samples were frozen in the presence of the best N-CPA either with 0.75mol/L ficoll, 0.2mol/L sucrose, 0.2mol/L trehalose or 0.05mol/L glycine. Sperm motility, membrane destabilization and viability were analysed to compare different freezing-thawing conditions. In addition, morphology and ultrastructure analysis were performed to compare fresh and frozen-thawed sperm quality. Our results indicate that the combination of EG and the thawing at 5°C improves (p≤.05) sperm post-thaw motility. Moreover, ficoll addition to EG-based freezing extender provided additional beneficial effect (p≤.05) on progressive movement and apoptosis incidence. Further work should evaluate different N-CPA concentrations to improve freezing protocol. In addition, fertility evaluation and testing on different chicken lines are needed in order to contribute to animal genetic resources bank.

48 EFFECT OF DIMETHYL SULFOXIDE- OR GLYCEROL-BASED VITRIFICATION PROTOCOLS ON THE DNA METHYLATION OF BOVINE CUMULUS-OOCYTE COMPLEXES

Although vitrification is becoming increasingly common for oocyte preservation, there has been recent evidence that some cryoprotectants may alter DNA methylation and so result in decreased oocyte developmental competence and abnormal embryonic development post-warming. The objective of this study was to determine the effect of dimethyl sulfoxide (DMSO)- or glycerol-based vitrification protocols on DNA methylation in bovine cumulus-oocyte complexes (COC). The vitrification protocols evaluated included a combination of ethylene glycol (EG) with either DMSO or glycerol (GLY). Cumulus-oocyte complexes were first exposed to equilibration solution (ES) consisting of 7.5% DMSO or GLY and 7.5% EG for 9 min at room temperature (RT) before being transferred to vitrification solution (VS) containing 15% DMSO or GLY, 15% EG, and 0.5 M sucrose. While in VS, 3 to 4 COC were loaded into an open system vitrification device (Cryolock®, BioTech Inc.) and plunged into LN within 1 min. For warming, COC were exposed to dilution solution 1 consisting of 0.5 M sucrose (37°C) for 2.5 min and to dilution solution 2 consisting of 0.25 M sucrose for 2.5 min (RT). Base media for all solution was PBS supplemented with 20% FBS. Cumulus-oocyte complexes (n = 137) were collected from crossbred cows by ovum pick-up on 3 separate days (repetitions), with half of the oocytes each day being matured to metaphase II (MII) stage before vitrification and the other half vitrified as germinal vesicle (GV) COC. The resulting treatments were DMSO GV v. Glycerol GV and DMSO MII v. Glycerol MII. Fresh COC (GV and MII stage, respectively) were utilised as controls. DNA methylation analysis of oocytes was performed through fluorescent staining using a primary antibody anti-5 mC (1:1000) and secondary antibody Alexa Fluor 488 (1:500). Fluorescence intensity of samples was measured using ImageJ software (NIH, Bethesda, MD, USA). Data were analysed through ANOVA with post hoc Tukey’s test. No differences between the vitrification groups or controls were found when analysing DNA methylation of GV stage COC (P = 0.1825). Metaphase II stage COC showed statistical difference between groups (P < 0.001) where glycerol-treated MII oocytes exhibited higher DNA methylation compared with DMSO MII and fresh MII COC. Results of the experiment showed that the glycerol-based vitrification protocol causes alteration of DNA methylation of bovine metaphase II cumulus-oocyte complexes, and as such may not be an optimal choice in mature oocyte vitrification protocols.

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

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

Experimental investigation on suitability of electrolytes for electrochemical micromachining of titanium

Titanium is a potential material for MEMS and biomedical uses owing to its unique high strength to weight ratio and their excellent corrosion resistance property. Machining of titanium by conventional as well as non-conventional methods is a challenge due to its inherent mechanical and chemical properties. Machining of titanium in micro domain is a topic of great research interest. Electrochemical micromachining (EMM) technique could be possible alternative for machining of titanium especially in micro domain. This paper presents systematic investigation of suitability of electrolyte for anodic dissolution of titanium through electrochemical micromachining. In order to investigate the performance of electrolytes, seven different types of electrolytes have been selected. The nature of dissolution of titanium and influence of electrolyte has been studied by machining a set of through micro holes in pure commercial titanium with all these different types of electrolytes. Five electrolytes out of seven have been demonstrated successful anodic dissolution of titanium using EMM process parameters. Out of these five electrolytes, two non-aqueous electrolytes were further evaluated, based on various criteria, i.e. material removal rate, radial overcut and conicity of micro hole. Amongst the two non-aqueous electrolytes, electrolyte with combination of ethylene glycol and sodium bromide has demonstrated excellent results during titanium micromachining by EMM.

Optimization of a vitrification protocol for hatched blastocysts from the dromedary camel (Camelus dromedarius)

The objective of this study was to modify and optimize a vitrification protocol (open pulled straw) that was originally designed for human oocytes and embryos, to make it suitable for the cryopreservation of camel hatched blastocysts. The original open pulled straw protocol was a complex process with 15-minute exposure of oocytes/embryos in 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (Me2SO) for equilibration, and cooling in 16% EG + 16% Me2SO + 1 M sucrose. Recognizing a need to better control the cryoprotectant (CPA) concentrations, while avoiding toxicity to the embryos, the effects on the survival rate and developmental potential of camel embryos in vitro were investigated using two different methods of loading the CPAs into the embryos (stepwise and semicontinuous increase in concentration), two different loading temperature/time (room temperature ∼24 °C/15 min and body 37 °C/3 min), and the replacement of Me2SO with EG alone or in combination with glycerol (Gly). A total of 145 in vivo–derived embryos were subjected to these processes, and after warming their morphological quality and integrity, and re-expansion was assessed after 0, 2, 24, 48, 72, and 96 hours of culture. Exposure of embryos in a stepwise method was more beneficial to the survival of embryos than was the semicontinuous process, and loading of CPAs at 37 °C with a short exposure time (3 minutes) resulted in an outcome comparable to the original processing at room temperature with a longer exposure time (15 minutes). The replacement of the Me2SO + EG mixture with EG only or a combination of EG + Gly in the vitrification medium significantly improved the outcome of all these evaluation criteria (P < 0.05). The modified protocol of loading EG at 37 °C for 3 minutes has increased the embryo survival of the original protocol from 67% to 91% and the developmental rate from 57% to 83% at 5-day culture. These results were comparable to or better than those reported in human or other species, indicating that this optimized method is well suited to any commercial embryo transfer program in the dromedary camel.

Experimental investigation of electrohydrodynamic instabilities in micro channels

An electric field is applied to destabilize the interface between two Newtonian and immiscible liquids flowing in a rectangular micro channel. The liquids are pumped into the micro channel with a syringe pump and a DC electric field is applied either parallel or normal to the flat interface between these liquids. The two liquids used in the experiments are a combination of ethylene glycol, different viscosity silicone oils, castor oil, and olive oil. The onset of electrohydrodynamic instability is investigated for various parameters, including the ratios of the flow rates, and viscosities of the liquids, the width of the micro channel, and the direction of the applied electric field. The order of the voltage applied to destabilize the interface is in the range 95 and 1190 V. The results of the experiments show that an increase in the viscosity ratio and the flow rate ratio of silicone oil to ethylene glycol have a stabilizing effect. It is also found that the important parameter to determine the critical voltage is the flow rate ratio, not the individual flow rates of the liquids. Also, as the width of the micro channel increases, the critical voltage increases. Lastly, for the liquid combinations used in the experiments, the interface could not be destabilized under the influence of a parallel electric field.

62 COMPARISON OF SUGARS, COMBINATIONS OF PERMEABLE CRYOPROTECTANTS, AND EQUILIBRATION REGIMENS FOR THE SOLID SURFACE VITRIFICATION OF IMMATURE PORCINE OOCYTES

Cryotop and solid surface vitrification are frequently used methods for the cryopreservation of porcine oocytes. These methods differ not only in the vitrification carrier but also in the cryoprotectant (CPA) treatment including the type of sugar, permeable CPA (pCPA) combinations, and the equilibration regimen. This study compared the distinct points of CPA treatment of these 2 methods to determine the optimum CPA treatment for the solid surface vitrification of immature porcine oocytes. We vitrified and warmed follicular cumulus-oocyte complexes by our method (Somfai et al. 2014 PLoS One 9, e97731). In each experiment, the vitrification solution consisted of 50 mg mL–1 polyvinyl pyrrolidone, 0.3 M of the actual sugar, and 35% [v/v] in total of the actual pCPA combination (depending on the experiment). After warming, the cumulus-oocyte complexes were subjected to in vitro maturation, IVF, and embryo culture (Kikuchi et al. 2002 Biol. Reprod. 66, 1033–1041). Oocyte survival was assessed after IVF by morphological evaluation, and live oocytes were subjected to in vitro embryo culture. Cleavage and blastocyst rates were calculated from cultured oocytes on Day 2 (Day 0 = IVF) and Day 6, respectively. Each experiment was replicated at least 3 times. Results were analysed by ANOVA. In Experiment 1, we compared trehalose (n = 416) and sucrose (n = 440) as supplementations during vitrification and warming (0.3 M and 0.4 M of each, respectively). There was no significant difference between oocytes vitrified with trehalose or sucrose in terms of survival, cleavage, and blastocyst development (83.2% v. 80.3%, 39.7% v. 42.4%, and 3.6% v. 5.9%, respectively). Thus, vitrification and warming media were supplemented with sucrose thereafter. In Experiment 2, we compared 1 : 1 combinations of ethylene glycol with propylene glycol (EG+PG group, n = 452) and ethylene glycol with dimethyl sulfoxide (EG+DMSO group, n = 465) used as pCPA for equilibration (4% [v/v] pCPA in total for 15 min) and vitrification (35% [v/v] pCPA in total for 30 s). Oocyte survival rate was higher (P &lt; 0.05) in the EG+PG group compared with the EG+DMSO group (73.8% v. 51.1%, respectively); however, cleavage and blastocyst development rates of surviving oocytes were not significantly different between the 2 groups (30.5% v. 44.5% and 4.1% v. 6.3%, respectively). In Experiment 3, we compared an equilibration treatment in 4% [v/v] of EG+PG for 13 to 15 min (regimen A, n = 368) with an equilibration in 15% [v/v] of EG+PG for 5 to 7 min (regimen B, n = 363) for oocyte vitrification. Survival, cleavage, and blastocyst development rates were higher (P &lt; 0.01) for oocytes vitrified using regimen A compared with those vitrified using regimen B (82.5% v. 22.7%, 24.0% v. 7.7%, and 3.2% v. 0%, respectively). In conclusion, trehalose and sucrose are equally effective during vitrification and warming, the combination of EG+PG as pCPA is superior to EG+DMSO, and equilibration in 4% pCPA for 13 to 15 min is superior to that in 15% pCPA for 5 to 7 min for the vitrification of immature porcine oocytes.This work was partly supported by JSPS KAKENHI Grant Number 26870839.