• The Journal of Advanced Prosthodontics
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• v.7 no.1
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• pp.21-26
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• 2015

PURPOSE. To evaluate the cytotoxicity of temporary luting cements on bovine dental pulp-derived cells (bDPCs). MATERIALS AND METHODS. Four different temporary cements were tested: Rely X Temp E (3M ESPE), Ultratemp (Ultradent), GC Fuji Temp (GC), and Rely X Temp NE (3M ESPE). The materials were prepared as discs and incubated in Dulbecco's modified eagle's culture medium (DMEM) for 72 hours according to ISO 10993-5. A real-time cell analyzer was used to determine cell vitality. After seeding $200{\mu}L$ of the cell suspensions into the wells of a 96-well plate, the bDPCs were cured with bioactive components released by the test materials and observed every 15 minutes for 98 hours. One-way ANOVA and Tukey-Kramer tests were used to analyze the results of the proliferation experiments. RESULTS. All tested temporary cements showed significant decreases in the bDPCs index. Rely X Temp E, GC Fuji Temp, and Rely X Temp NE were severely toxic at both time points (24 and 72 hours) (P<.001). When the cells were exposed to media by Ultratemp, the cell viability was similar to that of the control at 24 hours (P>.05); however, the cell viability was significantly reduced at 72 hours (P<.001). Light and scanning electron microscopy examination confirmed these results. CONCLUSION. The cytotoxic effects of temporary cements on pulpal tissue should be evaluated when choosing cement for luting provisional restorations.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.11
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• pp.1501-1508
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• 2004

In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.

• Korean Journal of Animal Reproduction
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• v.7 no.1
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• pp.19-23
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• 1983

Present studies were conducted to investigate the developmental stage and the location of embryos in the reproductive tract at various times after ovulation, the morphologically normal after thawing of embryos preserved in liquid nitrogen, and the survival after transferring frozen-thawed embryos. The results obtained were as follows: 1. Embryo stage and location in the reproductive tract after hCG administration. For the investigation of embryo stage and location in the reproductive tract after ovulation, rabbits were laparotomized at 24, 40, 48, 72 and 120 hrs post hCG injection, simultaneously with mating. the oviducts and uteri were flushed out with PBS medium containing 50% rabbit serum, respectively. 1) Most of embryos was remained in the oviduct within 48 hrs, with the lapse of time, embryos were started to move to uterus and shifted in uterus at 72 hrs after hCG injection. 2) The representatives of embryos stage collected at 24, 40, 48, 72 and 120 hrs were 1-cell(60.4%), 8-cell to early morula (52.3, 39.3%), late blastocyst (95.5%) stages, respectively. 2. Morphological normality and survival of the frozen-thawed embryos. For the evalution of the quality and viability on the frozen-thawed embryos, immediately after thawing, embryos were assessed by morphologically normal under a dissecting microscope, and a further test of frozen-thawed embryos was made by transferring the morphologically normal embryos to the uteri of recipient rabbit induced pseudopregnancy by the injection of hCG at the time of hCG injection in donor rabbits. 1) The propotions of embryos which a, pp.ared morphologically normal was higher when 8-cell (85.7%) and morula(90.5%) were used for freezing than when 4-cell (66.7%) and blastocyst (75.8%) were used. 2) Preganacies were observed at Day 15 after transfer of frozen-thawed 8-cell (7/13), morula (19/42) and blastocyst (3/19) but not after transfer of embryos at 4-cell stage.

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.115-118
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• 2008

Glycerol is the cryoprotectant most frequently used to freeze semen in several of species. The objective of the present study was to compare the effect of three different glycerol concentrations (4, 6 or 8%, v/v) on frozen-thawed dog sperm survival rate. Ejaculates from 9 dogs collected by digital manipulation were pooled and assessed by macroscopic and microscopic criteria. Semen was divided into 3 aliquots, which were centrifuged and the sperm pellets rediluted with first Tris-glucose-citric acid extender. After 1 h cooling at $4^{\circ}C$, second extender containing 4, 6 or 8% glycerol was added, respectively. The semen was loaded into 0.25 ml straws and frozen and stored in liquid nitrogen and thawed. Sperm vigor, live:dead spermatozoa ratio using HOS test, and sperm morphology using $Spermac^{(R)}$ stain were evaluated. After thawing, there were no significant differences among groups in vigor, viability and morphology. In conclusion, the three glycerol concentrations (4, 6 or 8%) can be used successfully in cryopreservation of canine semen. Therefore the use of 4% glycerol in the extender has less toxic effect and reduces of freezing injuries.

• Korean Journal of Veterinary Research
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• v.52 no.2
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• pp.89-97
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• 2012

Mycobacterial cell-wall skeleton (CWS) is an immunoactive and biodegradable particulate adjuvant and has been tried to use for immunotherapy. The CWS of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) was studied as an universal vaccine vehicle for antigen conjugation, to develop potentially effective and safe vaccine. Although a variety of biological activities of BCG-CWS have been studied, the effects of BCG-CWS on spleen cells are not fully elucidated. Using MTT assay and trypan blue exclusion test, we found that BCG-CWS significantly enhanced the viability and proliferation of cells. Multiple clusters, indicating proliferation, were observed in BCG-CWS-treated spleen cells and surface marker staining assay revealed that BCG-CWS promoted the proliferation of $CD19^+$ B lymphocyte rather than $CD4^+$ or $CD8^+$ T lymphocyte. In addition, BCG-CWS up-regulated the expression of anti-apoptotic molecules such as bcl-2, bcl-xL. BCG-CWS increased the surface expression of CD25 and CD69 as well as IL-2 production of spleen cells, suggesting increased activation. Furthermore, BCG-CWS enhanced the antigen-specific cell proliferation and interferon-gamma production of spleen cells. Taken together, these results demonstrate the immunostimulatory effects of BCG-CWS on spleen cells via multiple mechanisms, providing valuable information to broaden the use of BCG-CWS in clinical and research settings.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.1-6
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• 2012

This study evaluated a sexed sperm ability to produce embryos by flow cytometer. Hanwoo bulls sperm were separated to X and Y sperm via Hoechst 33342 stained with near UV laser or performed the pre-sorted without near UV laser beam in flow cytometry. Pre-sorted sperm had significantly higher viability ($84{\pm}1.15%$, $p$<0.05) compared to other sorted groups in frozen-thawed semen. For fresh semen, pre-sorted sperm had the higher viability ($79{\pm}3%$, $p$<0.05) than those of the X and Y sperm ($44.7{\pm}1.67$ and $41.7{\pm}1.2%$) separated by differences of DNA content. On the other hand, pre-sorted and X sperm sorted according to differences in DNA content had significantly higher viabilities ($24.3{\pm}1.2$ and $25.7{\pm}0.9%$, $p$<0.05) compared to that of the sorted Y sperm ($13.7{\pm}1.2%$) in the hypoosmotic swelling test. The proportion acrosome reaction in the sorted X sperm was higher ($55.0{\pm}1.7$ and $45.0{\pm}1.5%$) than those of the sorted Y-sperm ($32.3{\pm}0.9%$, $p$<0.05). However, the sperm morphologies of the sorted groups were not significantly differences. In conclusion, the sex-sorting procedure by flow cytometry affected some characteristics of Hanwoo sperm. Further study is needed to determine the optimal procedures to enhance male and female embryos and sorting accuracy.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.3
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• pp.247-257
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• 2014

Oil-in-water nanoemulsions of astaxanthin prepared by new vesicle, glyceryl citrate/ lactate/ linoleate/ oleate, were evaluated thoroughly in terms of cosmeceutical properties such as antioxidant effect, cell viability, influence of protein related enzyme, skin penetration, skin hydration and elasticity. Antioxidant effect and cell viability of nanoemulsion of astaxanthin were evaluated by DPPH and MTT assay. Also other properties of nanoemulsions of astaxanthin were measured by proteome analysis using 2D-PAGE, confocal laser scanning microscope and in-vivo test. We were able to find that the nanoemulsion of astaxanthin is good at scavenging of radical and inhibits the degradation of dermal extracellular matrix with the down-regulation of MMPs and other proteins related to MMP expression. CLSM was adopted for observing penetration of nanoemulsion of astaxanthin and showed high effective penetration rate compared to the nanoemulsion of astaxanthin prepared by conventional lecithin. In-vivo measurement of the nanoemulsions in hydration and elasticity were conducted to 11 Korean female adults for 28 days and showed better results.

• YAKHAK HOEJI
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• v.45 no.5
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• pp.536-544
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• 2001

increase the viability of oral typhoid vaccine during the passage through the castro-intes-tidal tract, numerous attempts have been made including the vaccine coating. However problems such as high death rate during the coating process and its instability in the gastric juice still remain to be solved. In this study, the oral vaccine was made as the micro-enteric beads by adding Salmomella typhi Ty21a cells to sodium alginate solution and spraying onto calcium chloride solution (ionotropic relation method). The vaccine showed more than 90% of its original viability after treating it for 1 hour in the artificial gastric juice (37$^{\circ}C$, 300 rpm). The clearance rate of the Ty21a in the liver and spleen of the mice orally administrated with coated Ty21a was similar to that of the mice intraperitoneally administrated with uncoated Ty21a. The peripheral blood lymphocytes (PBL) isolated from the mice orally administered with this vaccine produced 15.5 fold higher specific IgA antibody titer than that from the control mice administerd with saline solution. furthermore, the mice treated with the coated Ty21a had higher survival rates (50~87%) than the control mice treated with saline solution (0~10%) in the intraperitoneal challenge test with wild type S. typhi Ty21a cells. These results suggest that the alginate-based coating technique is effective to protect live Ty21a from acidic environments, and produces better intestinal immune responses thereby providing a potentially excellent oral typhoid vaccine.

• Journal of Veterinary Clinics
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• v.34 no.2
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• pp.156-160
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• 2017

We investigated the effect of ethylene glycol and antioxidants such as taurine, hypotaurine and trehalose with extenders during cryopreservation of Korean Jeju Black Bull spermatozoa. The cryopreservation of freshly collected spermatozoa was conducted with four different conditions. As a control, spermatozoa were cryopreserved with Tris egg-yolk extenders added 5% ethylene glycol (EG). Taurine (20 mM), hypotaurine (20 mM) and trehalose (20 mM) were individually added into tris egg-yolk extenders with 5% EG. After thawing of frozen spermatozoa with four different conditions, sperm viability, motility, acrosomal integrity, and membrane integrity were investigated. The significant (p < 0.05) improvement of sperm viability showed in all antioxidant treated thawed spermatozoa (taurine; $68.1%{\pm}4.4$, hypotaurine; $69.2%{\pm}6.7$ and trehalose; $68.0%{\pm}4.4$) when compared to control ($63.4%{\pm}5.6$). Neither positive nor detrimental effects of three antioxidants were shown sperm motility after thawing. The results of hypo-osmotic swelling test showed that the membrane integrity of taurine, hypotaurine or trehalose treated thawed spermatozoa ($64.1%{\pm}5.4$, $61.5%{\pm}3.7$ and $59.0%{\pm}4.0$, respectively) had significantly (p < 0.05) higher rate of the swollen sperm compared to control ($53.7%{\pm}9.7$). Hypotaurine treated frozen-thawed spermatozoa had siginificantly higher (p < 0.05) F pattern ratio than taurine, trehalose and control treated frozen-thawed spermatozoa. Trehalose added frozen-thawed spermatozoa had significantly higher (p < 0.05) acrosome reaction pattern ratio than taurine and hypotaurine added frozen-thawd spermatozoa. In this study, we found that antioxidants such taurine, hypotaurine and trehalose treatments during cryopreservation process could reduce damage of spermatozoa of Korean Jeju Black Bull and improved sperm capability of fertilization.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.1
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• pp.20-24
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• 2006

This study was designed to investigate the protective effect of Sanyakbojungbangam-tang Ethanol Extracts (SB Et-OH) on the cisplatin-induced apoptosis of human endothelial cell line ECV304 cells. After cells were treated with cisplatin, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, we used the several measures of apoptosis to determine whether this processes was involved in cisplatin-induced cell damage in ECV304 cells. Also, cells were treated with SB Et-OH and then, followed by the addition of cisplatin. Cisplatin decreased the viability of ECV304 cells in a dose-dependent manner and increased the caspase-3 enzyme activity ECV304 cells treated cisplatin were revealed as apoptosis characterized by nuclear staining. SB Et-OH protected ECV304 cells from cisplatin-induced nuclear fragmentation and chromatin condensation. Also, SB Et-OH inhibited the activation of caspase-3 pretense and the cleavage of poly(ADP-ribose) polymerase (PARP) in cisplatin-treated ECV304 cells. According to above results, SB Et-OH may protect ECV304 cells from the apoptosis induced by cisplatin.