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Improvement of Pregnancy Rates by Coculture of Human Embryos with Cumulus Cells in Glucose and Phosphate Free M-TALP Media (Glucose와 Phosphate가 제거된 M-TALP 배지에서의 난구세포 공배양에 의한 임신율 향상에 관한 연구)

  • Chung, B.S.;Chang, W.H.;Lee, M.H.;Kim, J.Y.;Bang, J.H.;Kim, K.H.;Suh, T.K.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.75-81
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    • 1999
  • The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

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Seismic Performance Evaluation of a Cone-type Friction Pendulum Bearing System (원추형 마찰진자베어링의 내진성능평가)

  • Jeon, Bub-Gyu;Chang, Sung-Jin;Park, Kyung-Rock;Kim, Nam-Sik;Jung, Duk-Young
    • Journal of the Earthquake Engineering Society of Korea
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    • v.15 no.2
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    • pp.23-33
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    • 2011
  • In this study, a CFPBS (Cone-type Friction Pendulum Bearing System) was developed which controls the acceleration delivered to the structure to prevent damage and degradation of the critical communication equipment in case of an earthquake. The isolation performance of the CFPBS was evaluated by numerical analysis. The CFPBS was manufactured in the shape of a cone differenced from the existing FPS (Friction Pendulum System), and a pattern was engraved on the friction surface. The natural frequencies of the CFPBS were evaluated from a free-vibration test with the seismic isolator system consisting of four CFPBSs. In order to verify its earthquake-resistant performance, a numerical analysis program was created from the equation of the CFPBS induced from the equations of motion. A simplified theoretical equation of the CFPBS was proposed to manufacture the equipment which could demonstrate the necessary performance. Artificial seismic waves satisfying the maximum earthquake scale of the Korean Building Code-Structural (KBC-2005) were created and verified to review the earthquake-resistant performance of the CFPBS by numerical analysis. The superstructural mass of the CFPBS and skew angle of the friction surface were considered for numerical analysis with El Centro NS (1940), Kobe NS (1995) and artificial seismic waves. The CFPBS isolation performance evaluation was based on the results of numerical analysis and the executed comparative analysis between the results from numerical analysis and the simplified theoretical equation under the same conditions.

Expression of Matrix metalloproteinase-1 between Simple Chronic Periodontitis and Type 2 Diabetes associated Chronic Periodontitis on Protein level (단순만성치주염환자와 2형 당뇨환자의 만성치주염에서 Matrix metalloproteinase-1의 발현양상)

  • Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
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    • v.35 no.3
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    • pp.649-659
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    • 2005
  • The purpose of this study was to quantify and compare the level of MMP-1 in the healthy or inflamed gingival tissue of patients with or without type 2 diabetic mellitus. We investigated whether mean amount of MMP-1 was changed by chronic periodontitis and type 2 DM. Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was divided into the three group. Group 1(n=8) was clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2(n=8) was inflamed gingiva from patients with chronic periodontitis. Group 3(n=8) was inflamed gingiva from patients with chronic periodontitis and type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantitative analysis of MMP-1 was performed using a densitometer and statistically analyzed by ANOVA. MMP-1 was expressed in all samples and an increased MMP-1 level was observed in group 2 compared to group 1 and decreased MMP-1 level was found group 3 compared to group 2, but the differences among 3 groups were not statistically significant. In conclusion, this study demonstrated that MMP-1 levels of inflamed gingiva of systemically healthy patient(group 2) were higher than normal gingiva of systemically health patients and although the severity of gingival inflammation in group 2 and 3 were similar, MMP-1 expression was decreased in diabetic patients than systemically healthy periodontal patients.

Citrus platymamma inhibits the expression of pro-inflammatory cytokines, inducible nitric oxide synthase, and cyclooxygenase-2 in RAW264.7 macrophage (RAW264.7 대식세포에서 Citrus platymamma의 iNOS, COX-2, 염증성 사이토카인 발현 억제 효과)

  • Kim, Sang Suk;Park, Kyung Jin;An, Hyun Joo;Choi, Young Hun
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1026-1032
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    • 2016
  • Citrus platymamma hort. ex Tanaka is widely used in traditional Korean medicine because of its medicinal benefits including an anti-inflammatory effect. This study aimed to evaluate changes in the flavonoid content and anti-inflammatory activities of C. platymamma during its harvest period. Fruit peel samples were obtained between September 2015 and February 2016. The results indicate that C. platymamma peel extract (CPE) was an effective inhibitor of lipopolysaccharide (LPS)-induced NO production in RAW264.7 cells. The inhibitory effects of CPE at $100{\mu}g/mL$ concentration included dose-dependent decreases in the expression of iNOS and COX-2 proteins. In addition, CPE decreased the expression of pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. The highest anti-inflammatory activity and flavonoid content were observed in CPE of C. platymamma peel harvested during the immature fruit period in September. Further, to assess the suitability of CPE for cosmetic use, we performed MTT assays using HaCaT keratinocytes and observed that CPE did not exhibit any cytotoxicity. To test the potential application of CPE as a cosmetic material, we also performed primary skin irritation tests on normal skin of 30 volunteers and no adverse reactions were observed. The results of this study indicate that CPE may be considered as an anti-inflammatory candidate for inclusion in cosmetic materials.

Bioequivalenee of Samchundang Atenolol Tablet to Tenolmin Tablet (테놀민 정에 대한 삼천당아테놀올 정의 생물학적동등성)

  • Cho, Hea-Young;Kang, Hyun-Ah;Lee, Suk;Baek, Seung-Hee;Lee, Yong-Bok
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.339-344
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    • 2003
  • Atenolol is a water soluble, ${\beta}_1$ selective adrenoceptor antagonist used in the treatment of angina and hypertension. It is primarily eliminated renally with minimal hepatic metabolism. The purpose of the present study was to evaluate the bioequivalence of Samchundang Atenolol (Samchundang Pharmaceutical Co., Korea.) to Tenolmin(Hyundai Pharmaceutical Ind. Co., Korea). The atenolol release from the two atenolol tablets in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty four normal male volunteers, 22.83$\pm$1.99 years in age and 65.82$\pm$7.15 kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 50 mg of atenolol was orally administered, blood was taken at predetermined time intervals and the concentrations of atenolol in serum were determined using HPLC method with fluorescence detector. The dissolution profiles of two atenolol tablets were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AUC_{t}$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_{t}$ and $C_{max}$ and untransformed $T_{max}$. The results showed that the differences in $AUC_{t}$, $C_{max}$ and $T_{max}$ between two tablets based on the Tenolmin were 3.74%, 4.38% and 17.77%, respectively. There were no sequence effects between two tablets in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(1.25) (e.g., log(0.98)∼log(1.l1) and log(0.95)∼log(1.l5) for $AUC_{t}$ and $C_{max}$ respectively), indicating that Samchundang Atenolol tablet is bioequivalent to Tenolmin tablet.

Regulation of stf Operon Expression and Its Virulence (살모넬라가 발현하는 stf 오페론의 조절과 병원성 인자로서의 기능)

  • Kim Sam-Woong;Kim Young-Hee;Kang Ho-Young
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.553-560
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    • 2005
  • The stf (Salmonella typhimurium fimbriae) operon consisting of stfA(CDEFG assumes to encode putative fimbriae. The complete stf operon is existed in S. typhimurium and S. choleraesuis, whereas it is absent in S. typhi. Analyses of the amino acid residues between major subunit StfA of the Stf fimbriae and those of known other fimbriaes suggested that Stf belongs to class I type fimbriae. Through comparison of StfD chaperone with the other fimbrial chaperones, and of C-terminus in subunits of Stf fimbriae, it belongs to FGS (with a short Fl-G1 loop) subfamily. In order to investigate the expression of stf operon, we have constructed a Salmonella strain containing a chromosomal stfA::lacZYA transcriptional fusion, resulting in S. typhimurium $_X8532$. The strain $_X8532$ lacked the expression of \beta-galactosidase$ under normal culture conditions. However, with longer incubation time of the S. typhimurium $_X8532$, we have isolated 21 individual strains exhibiting $Lac^+$ phenotype. $Lac^+$ phenotype was appeared as approximately 0.03 frequency per generation. All isolates expressed lacZ constitutively in the various environmental conditions. Various global regulatory proteins including RpoS, OmpR, and CpxR were not involved in the regulation of the stf operon. A S. typhimurium $_X8661$ mutant lacking stfAC function attenuated 6.7 folds more than that of wild type $_X3761$ in the mouse virulence test, suggesting in the somehow involved in the Salmonella pathogenesis.

Effect of Equilibration Time and Cell Stage on the Survival of IVF Bovine Embryos Cryopreserved by Vitrification (한우 체외수정란의 동결보존시 평형시간과 배 발달단계가 생존성에 미치는 영향)

  • 공일근;주영국;이은봉;김용권;박충생
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.7-14
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    • 1994
  • The present experiments on cryopreservation were designed to examine the effects of solution toxicity, equilibration time and cell stages on the post-thaw survival of bovine IVF embryos. The oocytes were matured in vitro(IVM) for 24 hrs. in TCM-199 supplemented with 35 $\mu$g /ml FSH, 10 $\mu$g /ml LH, 1 $\mu$g /ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro(IVC) with bovine oviductal epithelial cells for 7 to 9 days. The bovine IVF embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen, and thawed rapidly. 1. after the bovine blastocysts were exposed to EFS solution for 2 min. at room temperature and then they were washed in 0.5 M sucrose solution and TCM-199, they were cultured to examined cryoprotectant induced injury during exposure, Most of the embryos(95.0%) developed to reexpanded blastocoels. However, when the exposure time was extended to 5 and 10 min, these development rates dropped dramatically in 5 min. (69.5%) and 10 min. (47.4%), respectively, 2. When the bovine IVF embryos were vitrified in EFS solution after the equilibration for 1 and 2 min. exposure, The embryos to have reexpanded blastocoels following thawing, washing and culture processes were found to he 82.6 and 73.9%, respectively. However, when the exposure time was extended to 3 min, this survival rate dropped to 18.2%. The optimal time for equilibration of bovine IVF blastocysts in EFS solution seemed to he 1~2 min. 3. When the bovine IVF embryos were equilibrated for 1 min. the significantly (P<0. 05) higher post-thaw survival rates were obtained from the embryos of blastocyst stage(81.3%) than morulae stage(5. 1%). The optimal cell stage for viterification with EFS solution proven to he blastocyst stage in bovine IVF embryos. 4. The number of blastomeres of blastocyst stage was examined with nuclear staining with Hoechst 33342 during 7 to 9 days post-insemination. The cell counts of frozen bovine IVF embryos were found significantly(P$\geq$7.5 and those of the fresh embryos 76.6$\geq$7. 1, which were cultured in the sarne period and conditions as frozen embryos.

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Effect of Fe on the High Temperature Oxidation of Ti-Al-Fe Alloys (Ti-Al-Fe계 합금의 고온산화거동에 미치는 Fe의 영향)

  • Yoon, Jang-Won;Hyun, Yong-Taek;Kim, Jeoung-Han;Yeom, Jong-Taek;Yoon, Seog-Young
    • Korean Journal of Materials Research
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    • v.21 no.7
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    • pp.357-363
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    • 2011
  • In this paper, high temperature oxidation behavior of newly developed alloys, Ti-6Al-4Fe and Ti-6Al-1Fe, is examined. To understand the effect of Fe on the air oxidation behavior of the Ti-Al-Fe alloy system, thermal oxidation tests are carried out at $700^{\circ}C$ and $800^{\circ}C$ for 96 hours. Ti-6Al-4V alloy is also prepared and tested under the same conditions for comparison with the developed alloys. The oxidation resistance of the Ti-Al-Fe alloy system is superior to that of Ti-6Al-4V alloy. Ti-6Al-4V shows the worst oxidation resistance for all test conditions. This is not a result of the addition of Fe, but rather it is due to the elimination of V, which has deleterious effects on high temperature oxidation. The oxidation of the Ti-Al-Fe alloy system follows the parabolic rate law. At $700^{\circ}C$, Fe addition does not have a noticeable influence on the amount of weight gain of all specimens. However, at $800^{\circ}C$, Ti-6Al-4Fe alloy shows remarkable degradation compared to Ti-6Al-1Fe and Ti-6Al. It is discovered that the formation of $Al_2O_3$, a diffusion resistance layer, is remarkably hindered by a relative decrease of the ${\alpha}$ volume fraction. This is because Fe addition increases the volume fraction of ${\beta}$ phase within the Ti-6Al-xFe alloy system. Activities of Al, Ti, and Fe with respect to the formation of oxide layers are calculated and analyzed to explore the oxidation mechanism.

Therapeutic Benefits of Mesenchymal Stromal Cells in a Rat Model of Hemoglobin-Induced Hypertensive Intracerebral Hemorrhage

  • Ding, Rui;Lin, Chunnan;Wei, ShanShan;Zhang, Naichong;Tang, Liangang;Lin, Yumao;Chen, Zhijun;Xie, Teng;Chen, XiaoWei;Feng, Yu;Wu, LiHua
    • Molecules and Cells
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    • v.40 no.2
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    • pp.133-142
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    • 2017
  • Previous studies have shown that bone marrow mesenchymal stromal cell (MSC) transplantation significantly improves the recovery of neurological function in a rat model of intracerebral hemorrhage. Potential repair mechanisms involve anti-inflammation, anti-apoptosis and angiogenesis. However, few studies have focused on the effects of MSCs on inducible nitric oxide synthase (iNOS) expression and subsequent peroxynitrite formation after hypertensive intracerebral hemorrhage (HICH). In this study, MSCs were transplanted intracerebrally into rats 6 hours after HICH. The modified neurological severity score and the modified limb placing test were used to measure behavioral outcomes. Blood-brain barrier disruption and neuronal loss were measured by zonula occludens-1 (ZO-1) and neuronal nucleus (NeuN) expression, respectively. Concomitant edema formation was evaluated by H&E staining and brain water content. The effect of MSCs treatment on neuroinflammation was analyzed by immunohistochemical analysis or polymerase chain reaction of CD68, Iba1, iNOS expression and subsequent peroxynitrite formation, and by an enzyme-linked immunosorbent assay of pro-inflammatory factors (IL-$1{\beta}$ and TNF-${\alpha}$). The MSCs-treated HICH group showed better performance on behavioral scores and lower brain water content compared to controls. Moreover, the MSC injection increased NeuN and ZO-1 expression measured by immunochemistry/immunofluorescence. Furthermore, MSCs reduced not only levels of CD68, Iba1 and pro-inflammatory factors, but it also inhibited iNOS expression and peroxynitrite formation in perihematomal regions. The results suggest that intracerebral administration of MSCs accelerates neurological function recovery in HICH rats. This may result from the ability of MSCs to suppress inflammation, at least in part, by inhibiting iNOS expression and subsequent peroxynitrite formation.

Selection of Promising 'Ssam' Vegetable for Summer Production in Highland (고랭지 여름출하용 유망 쌈채소 선발)

  • Jang, Suk-Woo;Kim, Won-Bae;Ryu, Kyoung-Ou
    • Horticultural Science & Technology
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    • v.19 no.2
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    • pp.140-144
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    • 2001
  • Field investigations were initiated to determine possible new 'ssam' vegetable that could be grown during the summer season in highland. Korean 'ssam' means cooked rice in vegetables. The 'ssam' vegetable is called that those leaves has been used for wrapping at meal. Seventeen leafy vegetables were studied, including chard (Beta vulgaris L. var. flavescens DC.), leaf broccoli (Brassica oleracea L. var. italica), Chinese leek (Allium tuberosum Rottl.), Japanese hornwort (Cryptotaenia japonica Hassk), red leaf mustard (Brassica juncea L.), green leaf mustard (Brassica juncea L.), leaf parsley (Petroselinum crispum Nijim), pakchoi (Brassica campestris L. ssp. chinensis Jusl.) tatsoi (Brassica campestris var. narisoma), kale (Brassica oleracea var. acephala), collard (Brassica oleracea var. acephala), Korean mint (Agastache rugosa O. Kuntze), romaine lettuce (Lactuca sativa L. var. longifolia Lam.), red leaf chicory (Cichorium intybus L. var. folisum), red chicory (Cichorium intybus L. var. folisum), green leaf chicory (Cichorium intybus L. var. folisum), and sprouting broccoli (Brassica oleracea L. var. italica). Date of the first harvest in 1999 was July 10th in chard, leaf broccoli, red mustard, mustard, leaf parsley, pakchoi, tatsoi, kale, collard, Korean mint, red leaf chicory, red chicory, green leaf chicory, and sprouting broccoli. Date of the first harvest for Chinese leek, Japanese hornwort, and romaine lettuce was from the middle of June to the late of August. Based on ratings on marketable yield, we found good leaf shape, taste and yield in chard, red leaf mustard, green leaf mustard, pakchoi, tatsoi, romaine lettuce, red leaf chicory, red chicory, and green leaf chicory. Even though the most of all these vegetables were obtained high scores by test panels in shape, taste and quality, but some of them revealed inadequate leaf size as a 'ssam' vegetable. That problem will be solved by control of harvesting time. On the basis of this observation, chard, red leaf mustard, green leaf mustard, pakchoi, tatsoi, romaine lettuce, red leaf chicory, red chicory, and green leaf chicory were rated as promising 'ssam' vegetable by analysis of profits, adaptation, and quality.

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