• Journal of Embryo Transfer
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• v.28 no.1
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• pp.49-55
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• 2013

The objective of this study was to evaluate several types of uterine bacteria in Hanwoo. uterine bacteria from randomly selected 5 uterus was collected by flushing methods into a sterilized 1.5 ml centrifuge tube and was inoculated onto MacConkey agar and blood agar, respectively. After being incubated for 5% $CO_2$, aerobic or anaerobic condition at $37^{\circ}C$ during 48h, bacterial colonies were selected and re-inoculated onto blood agar plates. Re-cultured colonies were identified by Gram staining and finally identified using Vitek system. The identified bacteria were Staphylococcus lentus, Staphylococcus sciuri, Staphylococcus vitulinus, Staphylococcus warneri of Gram (+) and Rhizobium radiobacter, Sphingomonas paucimobilis of Gram (-) bacteria. Although, pathogenicity of identified bacteria was unclear, the bacteria can have an effect on the uterine microenvironment. Therefore, repetitive research will be required to determine the effects of bacteria in cattle exposed to a various environment.

• Archives of Plastic Surgery
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• v.36 no.6
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• pp.679-684
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• 2009

Purpose: Human lipoaspirate cells are relatively easy to obtain in large quantities without cell culture. The aim of this in vitro pilot study was to determine the effect of cell therapy using uncultured lipoaspirate cells on cell proliferation and collagen synthesis of diabetic fibroblasts, which are the major contributing factors in wound healing. Methods: In order to get diabetic fibroblasts, dermis tissues were obtained from foot skin of diabetic patients who underwent debridements or toe amputations(n = 4). In order to isolate lipoaspirate cells, the same diabetic patients' abdominal adipose tissues were obtained by liposuction. The diabetic fibroblasts were co - cultured with or without autogenous lipoaspirate cells using porous culture plate insert. Initial numbers of the lipoaspirate cells and diabetic fibroblasts seeded were 15,000 cells/well, respectively. For cell proliferation assay, two treatment groups were included. In group I, diabetic fibroblasts were cultured with the insert having no cells, which serves as a control. In group II, the lipoaspirate cells were added in the culture plate insert. For collagen synthesis assay, one additional group(group III), in which diabetic fibroblasts were not seeded in the well and only lipoaspirate cells inside the insert were incubated without diabetic fibroblasts, was included for a reference. Results: One hundred to one hundred sixty thousand lipoaspirate cells were isolated per ml of aspirated adipose tissue. After 3 - day incubation, the mean cell numbers in group I and II were 17,294/well and 22,163/well. The mean collagen level in group I, II, and III were 29, 41, and 2 ng/ml, respectively. These results imply that both cell proliferation and collagen synthesis in the lipoaspirate cell treatment group were 28 and 44 percents higher than in the control group, respectively(p < 0.05). Conclusion: Uncultured lipoaspirate cell autografts may stimulate the wound healing activity of diabetic fibroblasts.

• Korean Journal of Veterinary Research
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• v.32 no.4
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• pp.663-669
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• 1992

This experiment was carried out to find out the best condition for the parthenogenetic activation of mouse eggs by treating ethanol and hyaluronidase. For the parthenogenetic activation of eggs with ethanol, cumulus cell enclosed or denuded eggs were treated with 7% ethanol in D-PBS for 5, 7 or 9 minutes. For the activation of eggs with hyaluronidase, the eggs with cumulus masses were released into D-PBS with 100 unit hyaluronidase and treated for 10, 12 or 13 minutes. All of the treated eggs were incubated in BMOC-3 solution for 5 hours at $37^{\circ}C$ at an atmosphere of 5% $CO_2$ in air. The types of parthenogenetic eggs were morphologically classified into haploid, diploid, immediate cleavage eggs under an inverted microscope. The results obtained in this experiment were summarized as follows ; 1. High activation rate(99%) had been achieved by treating the eggs with 7% ethanol for 7 minutes. 2. With 100 IU hyaluronidase, high activation rate (94%) had been achieved by treating for 12 minutes. 3. The most frequent type of parthenogenetic eggs activated with ethanol or hyaluronidase was haploid (p<0.05). 4. The eggs collected from 18 to 22 hours post HCG injection showed higher activation rate than the eggs collected at 16 hours post HCG injection. 5. No significant difference (p>0.05) in activation rate was shown in strain of mouse and in presence of cumulus cells.

• Journal of The Korean Society of Agricultural Engineers
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• v.56 no.6
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• pp.169-176
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• 2014

To evaluate bio-cementation of microbial on sands, laboratory test was conducted using acrylic cubic molding boxes ($5cm{\times}5cm{\times}5cm$). It was incubated the microbial, called Bacillus Pasteurii, according to Park et al (2011, 2012). and applied 50ml each specimen. Two type of sand samples used were Jumoonjin sand and common sand (well graded). These sands were molded in acrylic boxes with the relative density of 30 % and 60 % respectively. Microbial were poured onto the samples molded in acrylic boxes and cured at the room temperature and humidity. After 7, 14 and 21days, it was measured the compressive strength, pH, EC, and density and it were observed SEM and XRD to verify the effect of bio-cementation. It was found that bio-cementation was increased a strength of sands and it was appeared that strengths were related to the type of sand and relative density. Therefore it was confirmed the solidification of sands using the bio-cementation by microbial activation and the usefullness of acrylic molding boxes when tests were conducted on the soil of sands.

• Journal of Pharmaceutical Investigation
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• v.28 no.2
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• pp.121-126
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• 1998

The solubility and physicochemical stability of caroverine hydrochloride (CRV), an antispasmodic, in buffered aqueous solutions were studied using a reverse phase high performance liquid chromatography. The solubilty of the drug at pH 2.76-5.40 was similar at the range 31.9-36.2 mg/ml $(34^{circ}C)$, but, at the pH higher than 6.0, markedly decreased. The use of polyethylene glycol 400 as a cosolvent did not increase the solubility at any compositions examined. Moreover. increasing molar concentration of aqueous phosphate buffer from 0 to 0.5 M remarkably decreased the solubility. The degradation of CRY followed the apparent first-order kinetics. The degradation was accelerated with decreasing pH and increasing storage temperature. The half-lives for the degradation of CRY (1.0 mg/ml) at pH 1.28. 4.01 and 5.93 $(45^{\circ}C)$ were 2.8, 31.4 and 124 hr. respectively. The pHs of incubated solutions were to some extent lowered perhaps due to the formation of acidic degradation products. The addition of disodium edetate (0.01%) to the CRY solution (pH 4.95) retarded 2.5 times the degradation rate at $45^{\circ}C$, but the use of sodium bisulfite (0.1%) accelerated 2.9 times the rate. The activation energy for the CRY solution (20 mg/ml. pH 5.4) containing 0.01% EDTA was calculated to be 5.98 kcal/mole. When the solution was stored under nitrogen displacement in ampoule, there was no significant degradation even after 3 months at $40^{\circ}C$, indicating that protection from oxidation by air (oxygen) is essential for the complete stabilization of CRY solution.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.106-110
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• 1995

An alkaline protease-producing bacterium was isolated from Korean hot pepper paste and identified as Alteromonas sp. CN301. A alkaline protease was purified and characterized. The optimal pH and temperature for the enzyme activity were pH 12.0 and $35^{\circ}C$, respectively. Molecular weight of the enzyme was determined as 31,000 dalton by the SDS-PAGE. The enzyme was stable in the range of $pH\;6.0{\sim}13.0$ showing the residual activity above 80% of the enzyme activity. The residual activity of the enzyme was 64% when the enzyme was incubated at $50^{\circ}C$ for 1 hr. The activity of the enzyme was not affected by most metal ions tested except $Hg^{2+}$, and activated by Triton X-100, Tween 20 and Tween 80. The enzyme activity was severely inhibited by PMSF and EDTA, suggesting that the enzyme is serine protease having metal ion in its structure.

• Korean Journal of Clinical Laboratory Science
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• v.39 no.1
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• pp.19-24
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• 2007

The aim of this study was to evaluate the bacterial effects of Moraxella catarrhalis in otitis media with effusion (OME) by photodynamic therapy (PDT). Bacterial suspensions (10000 CFU/mL) were prepared. The colony forming units (CFU) of Moraxella catarrhalis have been measured after an application of photogem plus 632 nm diode laser irradiation. One ml of the bacterial suspensions have been incubated in the dark for 3h with various concentrations of photogem ($0.625{\sim}5.0_{\mu}g/mL$) and then irradiated with 632 nm diode laser ($15J/cm^2$). After, the PDT Moraxella catarrhalis suspensions ($50{\mu}L$) were inoculated on chocolate agar plate and cultured in the dark at $37^{\circ}C$, 5% $CO_2$ condition for 18h. The colony forming units off the bacteria were measured. Also transmission electron microscopy (TEM) was employed to evaluate the effect of otitis media pathogens by PDT. The nucleus of Moraxella catarrhalis was stained using green fluorescent nucleic acid dye thiazole orange and the fluorescence intensity of the nucleus was measured by flow cytometry. The PDT was effective in killing Moraxella catarrhalis at the photogem dose of $5.0_{\mu}g/mL$, respectively, As assessed by flow cytometry analysis the fluorescence intensity of the nucleus got lower after PDT. TEM result appeared to able to cause damage to the bacterial membranes. On the basis of these findings, bacterial photodynamic therapy with photogem can be considered to be a promising new therapeutic approach for OME.

• Nutrition Research and Practice
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• v.11 no.2
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• pp.83-89
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• 2017

BACKGROUND/OBJECTIVES: Epithelial-mesenchymal transition (EMT) is involved in not only cancer development and metastasis but also non-cancerous conditions. Hypoxia is one of the proposed critical factors contributing to formation of chronic rhinosinusitis or nasal polyposis. Wheatgrass (Triticum aestivum) has antioxidant, anti-aging, and anti-inflammatory effects. In this study, we analyzed whether wheatgrass has an inhibitory effect on the EMT process in airway epithelial cells. MATERIALS/METHODS: A549 human lung adenocarcinoma cells were incubated in hypoxic conditions ($CO_2$ 5%/$O_2$ 1%) for 24 h in the presence of different concentrations of wheatgrass extract (50, 75, 100, and $150{\mu}g/mL$) and changes in expression of epithelial or mesenchymal markers were evaluated by immunoblotting and immunofluorescence. Accordingly, associated EMT-related transcriptional factors, Snail and Smad, were also evaluated. RESULTS: Hypoxia increased expression of N-cadherin and reduced expression of E-cadherin. Mechanistically, E-cadherin levels were recovered during hypoxia by silencing hypoxia inducible factor (HIF)-$1{\alpha}$ or administering wheatgrass extract. Wheatgrass inhibited the hypoxia-mediated EMT by reducing the expression of phosphorylated Smad3 (pSmad3) and Snail. It suppressed the hypoxia-mediated EMT processes of airway epithelial cells via HIF-$1{\alpha}$ and the pSmad3 signaling pathway. CONCLUSION: These results suggest that wheatgrass has potential as a therapeutic or supplementary agent for HIF-1-related diseases.

• Journal of Embryo Transfer
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• v.24 no.2
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• pp.121-125
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• 2009

In the present study, effects of concentration of cryoprotectant solutions on the nuclear maturation of vitrifiedthawed bovine oocytes were examined. Also, the developmental capacity of vitrified-thawed immature oocytes following ICSI was investigated. Oocytes were cultured in TCM-199 medium supplemented with 5% FBS at $38^{\circ}$C in 5% $CO_2$ and air. The in vitro maturation rate of vitrified oocytes was 24.5 ${\pm}$ 4.2%. The in vitro maturation rate of vitrified oocytes was lower than that of the control (72.0 ${\pm}$ 3.5%, p<0.05). The in vitro maturation rate of vitrified${\sim}$thawed oocytes incubated in TCM-199 medium supplemented with 1.0${\sim}$5.0 ug CB were 26.7 ${\pm}$ 3.2%, 35.7 ${\pm}$ 3.2%, 54.0 ${\pm}$ 3.0%, 42.5 ${\pm}$ 3.6%, respectively. The in vitro maturation rate (57.0 ${\pm}$ 3.0%) of the vitrified-thawed oocytes treated with 3.0 ${\mu}$g CB for 20 min was the highest of all vitrification groups, although the maturation rate were significantly (p<0.05) lower than those of fresh oocytes. The in vitro maturation rates of the vitrified-thawed (with EDS and EDT) oocytes were 53.8 ${\pm}$ 3.4%, 51.1 ${\pm}$ 3.5%, respectively. This results were lower than the control group (72.0 ${\pm}$ 3.0%). The in vitro developmental rates of the vitrified-thawed oocytes following ICSI were 28.6 ${\pm}$ 4.5%, 25.6 ${\pm}$ 4.3%, respectively. This results were lower than the control group (40.0 ${\pm}$ 4.0%).

• Journal of Biomedical Engineering Research
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• v.32 no.3
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• pp.185-190
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• 2011

The purpose of this study is to investigate the influence of organism vigor information solution(OVIS) on the animal cell growth and to set up a condition for cell culture. We investigated the reactions on the MG-63 and MCF-7 cell line in mixture culture media with various amount of REVIEW solution, which is an example among various OVIS. DMEM-HG was used as basic media. The concentration range of the mixture was limited from 0% to 15%. MTT assay is used for cell viability test. The cell was incubated for 14days and the MTT assay was performed on day 1, 3, 7, 10 and 14 throughout the experiment. We used the ELISA reader to measure the Optical Density(O.D) at 595 nm wavelength filter. MCF-7 was linearly proliferated according to culture time and concentrations of OVIS. On the other hand, MG-63 cells were measured the highest O.D value at 12%. The growth rates of both cells cultured in mixed culture media with OVIS are much higher than those in only basic media, DMEMHG, after 14days. It was confirmed that cell cultured at OVIS grows rapidly at certain period although cells showed a negative effect in initial stage.