• Title/Summary/Keyword: Cell treatment

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Effect of angiotensin II inhibition on the epithelial to mesenchymal transition in developing rat kidney (발생 중인 백서 신장에서 Angiotensin II 억제가 epithelial to mesenchymal transition에 미치는 효과)

  • Yim, Hyung-Eun;Yoo, Kee-Hwan;Bae, In-Sun;Hong, Young-Sook;Lee, Joo-Won
    • Clinical and Experimental Pediatrics
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    • v.52 no.8
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    • pp.944-952
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    • 2009
  • Purpose : To investigate the effects of angiotensin II inhibition on the epithelial to mesenchymal transition (EMT) in the developing kidney, we tested the expression of EMT markers and nestin in angiotensin converting enzyme (ACE) inhibitor-treated kidneys. Methods : Newborn rat pups were treated with enalapril (30 mg/kg/d) or a vehicle for 7 days. Immunohistochemistry for the expression of ${\alpha}$-smooth muscle actin (SMA), E-cadherin, vimentin, and nestin were performed. The number of positively-stained cells was determined under 100 magnification in 10 random fields. Results : In the enalapril-treated group, ${\alpha}SMA-positive$ cells were strongly expressed in the dilated tubular epithelial cells. The number of ${\alpha}SMA-positive$ cells in the enalapril-treated group increased in both the renal cortex and medulla, compared to the control group (P<0.05). The expression of E-cadherin-positive cells was dramatically reduced in the cortical and medullary tubular epithelial cells in the enalapril-treated group (P<0.05). The number of vimentin- and nestin-positive cells in the cortex was not different in comparisons between the two groups; however, their expression increased in the medullary tubulointerstitial cells in the enalapril-treated group (P<0.05). Conclusion : Our results show that ACE inhibition in the developing kidney increases the renal EMT by up-regulating ${\alpha}SMA$ and down-regulating E-cadherin. Enalapril treatment was associated with increased expression of vimentin and nestin in the renal medulla, suggesting that renal medullary changes during the EMT might be more prominent, and ACE inhibition might differentially modulate the expression of EMT markers in the developing rat kidney.

Clinical Significance of Pyuria in Pediatric Patients with Febrile Urinary Tract Infection (발열성 소아 요로감염에서 농뇨의 임상적 의의)

  • Choi, Lim;Cho, Sea-Eun;Yim, Hyung-Eun;Yoo, Kee-Hwan;Hong, Young-Sook;Lee, Joo-Won
    • Childhood Kidney Diseases
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    • v.15 no.1
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    • pp.66-75
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    • 2011
  • Purpose: Early diagnosis and treatment of febrile urinary tract infection (UTI) in children is important to prevent kidney damage. This study aims to evaluate the relationship between the presence of pyuria, the severity, and underlying genitourinary anomalies in patients with UTI. Methods: We retrospectively reviewed 293 patients with febrile UTI who were admitted to Korea University Guro Hospital during the period from June, 2007 until January, 2010. We divided the patients into two groups, one with the finding of pyuria at admission, and the other without, and compared the fever duration, white blood cell counts (WBC) and C-reactive protein (CRP) in peripheral bloods, hydronephrosis, cortical defects, vesicoureteral reflux and admission period. Results: Among the 293 patients with febrile UTI, 189 patients showed findings of pyuria whereas 104 patients did not. Patients with pyuria showed an increment of WBC ($14,694{\pm}485.2$ vs. $11,374{\pm}451.2/uL$, P <0.05) and CRP ($46.9{\pm}3.9$ vs $17.1{\pm}3.6$ mg/L, P <0.05) in peripheral blood sample. The presence of cortical defects (21.7 Vs 5.8%, P <0.05) and vesicoureteral reflux (15.9 Vs 6.7%, P <0.05) was also increased in patients with pyuria compared to patients without pyuria. There were no specific differences in fever duration, admission period, and hydronephrosis. Within the group with pyuria, CRP in peripheral blood sample increased proportionally with the increment of pyuria (P <0.05). Conclusion: In patients with febrile UTI, the increment of WBC in the urine sample can be a helpful predictor for increased CRP in peripheral blood and acute pyelonephritis.

Lung Preservation Study for Above 20 Hours of LPDG Solution in Canine Lung Allotransplactation (폐이식 실험견에서 LPDG용액을 이용한 20시간 이상 폐보존효과 관찰)

  • Park, Chang-Gwon;Gwon, Geon-Yeong;Yu, Yeong-Seon
    • Journal of Chest Surgery
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    • v.30 no.10
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    • pp.949-960
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    • 1997
  • Background. Limited ischemic tolerance of the lung has remained one of the factors that limits the expansion of pulmonary transplantation as a treatment for end-stage pulmonary disease. Numerous studies on safe long term preservation for lung transplantation has been performed for the purpose of developing ideal preservation solution with extracellular type or intracellular type solutions. In this. study, we examined the efficacy of L DG solution in lung preservation longer than 20 hours by comparison with modified Euro-Collins solution. Iwethods. Thirty-(our adult mongrel dogs were divided into two groups. Donor lungs were flushed with LPDG solution(n=9) or modified Euro-Collins(MEC) solution(n=8) and stored for 24 hours at 1$0^{\circ}C$. All donor lungs were perfused through the pulmonary arteries with solutions containing prostaglandin El and verapamil. Left canine lung allotransplantations wereperformed. Assessment(hemodynamic indices and arterial blood gas analysis) of left implanted lung was made by occluding the right pulmonary artery for ten minutes using pulmonary artery Cuff. Assessment was repeated at the interval of 30 minutes, one hour, and two hours later after reperfusion and then chest X-ray, computed tomogram and lung perfusion scan were obtained. In survival dogs follow-up studies were done with assessment with chest X-ray, computed tomogram of the chest and lung perfusion scan on 7th day postoperatively. After preservation above 20 hours, pathological examinations for ultrastructural findings on right lung were performed in each group. Results. With respect to arterial oxygen tension, LPDG group was superior to MEC but there was no statistical significance for 2 hours after reperfusion. Mean pulmonary artery pressure was less increased(p < 0.05) and cardiac output higher(p <0.05) than MEC group until 2 hours after reperfusion. After 2 hours of reperfusion, both groups showed transplanted lung function deteriorated gradually. Perfusion scan of the transplanted lung in LPDG group showed better perfusion rate in immediate post-reperfusion, 3 days and 7 days later respectively but there was no statistical significance and corelation with PaO2 and computed tomoRravhic views. In scanning electron microscopy of pulmonary artery after preservation, LPDG group relatively shows less irregular protrusion of the inner surface of endothelial cell of poulmonary artery than MEC group. Conclusions, e concluded that LPDG solution can offer safe lung preservation above 20 hours with adequate immunosuppressive therapy and prevention of the infection.

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Chondrogenesis of Human Adipose Tissue Derived Mesenchymal Stem Cells (ATMSCs) Seeded in Gelatin-Chondroitin-Glucosamine Scaffold (Gelatin-Chondroitin-Glucosamine Scaffold에 접종한 인간지방조직-유래 중간엽 줄기세포의 연골형성)

  • Kim, Eung-Bae;Hong, Soon-Gab;Do, Byung-Rok;Kim, Hae-Kwon;Lee, Joon-Yeong
    • Development and Reproduction
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    • v.15 no.2
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    • pp.99-111
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    • 2011
  • The present experiment was performed to evaluate the chondrogenic differentiation potential of human adipose tissue-derived mesenchymal stem cells (ATMSCs) in the chondrogenic induction medium (CIM) with transforming growth factor-${\beta}1$ (TGF-${\beta}1$) and to evaluate the chondrogenic differentiation of ATMSCs seeded in gelatin-chondroitinglucosamine scaffold (GCG-scaffold). ATMSCs and mouse chondrocytes were cultured in the basic medium and CIM without TGF-${\beta}1$ (CIM1) or with TGF-${\beta}1$ (CIM2) for chondrogenic differentiation potential. The chondrogenic differentiation of ATMSCs was evaluated by glycosaminoglycan (GAG) synthesis and histochemical staining. In pellet culture, GAG synthesis of ATMSCs and chondrocyte was increased in culture on 14 days, but higher in CIM1 than basic medium, especially highest in CIM2. Cartilage matrix was observed in ATMSCs cultured in CIM2 on 14 days by Safranin O and trichrome staining. In well plate culture, proliferation of ATMSCs was continuously increased in culture on 10 days and higher in CIM than basic medium. The cell adhesion rate of ATMSCs seeded in flask or scaffolds was continuously increased during culture period, but higher in scaffold than flask. GAG synthesis of ATMSCs seeded in scaffolds showed no change in control group. In the CIM groups, GAG synthesis of ATMSCs was continuously increased than control group during culture period, especially very high in CIM2 and in the GCG-scaffold was slightly higher than the gelatin scaffold (G-scaffold). The present results demonstrated that ATMSCs showed an low chondrogenic differentiation potential, compared to mouse chondrocytes for 14 days of culture. TGF-${\beta}1$ is important factor in chondrogenic differentiation of ATMSCs. Gelatin scaffold was considered to increasing the effective chondrogenic differentiation environment. ATMSCs seeded in GCG-scaffold was more effective in chondrogenesis than in G-scaffold. Conclusively, the present results demonstrated that the treatment of chondroitin and glucosamine in the scaffold was more effective to promote the cartilage matrix formation.

Effect of Ethane 1,2-Dimethane Sulfonate (EDS) on the Accessory Sex Organs in Adult Rats : A Histological Study (Ethane 1,2-Dimethane Sulfonate(EDS)가 성체 흰쥐의 부속 생식기관에 미치는 효과 : 조직학적 연구)

  • Lee, Won-Yong;Lee, Sung-Ho
    • Development and Reproduction
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    • v.13 no.2
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    • pp.105-114
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    • 2009
  • Ethane 1,2-dimethane sulfonate (EDS) is a well-known alkylating agent used as selective Leydig cell (LC) toxicant to create a testicular dysfunction model. Previous studies including our own clearly demonstrated the dramatic weight loss of the androgen dependent accessory sex organs such as epididymis, seminal vesicle and prostate gland in this 'LC knock-out' rats. The present study was performed to evaluate the effect of EDS administration on histological changes of the epididymis, seminal vesicle and prostate in adult rats. Adult male Sprague-Dawley rats (350$\sim$400 g B.W.) were injected with a single dose of EDS (75 mg/kg, i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. Tissue weights (testis, epididymis, seminal vesicle and prostate gland) were measured. The histological changes of tissue were observed by a light microscopy using hematoxylin & eosin staining. Weights of the reproductive and accessory organs progressively declined after the EDS treatments (weeks 1, 2 and 3). After this, the decrease was stopped, then gradually returned to the normal levels. There was a partial (about 60%) recovery of the epididymis weight during weeks $6{\sim}7$. The cross section of epididymis revealed an increase in thickness of the epithelium during weeks $1{\sim}3$. In contrast, considerable reduction of epithelial thickness in seminal vesicle was observed during same period. Similarly, a reduction in thickness of prostate epithelial layer was found during weeks $1{\sim}3$, then it was back to normal thickness after week 4. Taken together, the present study demonstrated that the temporally induced androgen-deficiency by EDS treatment could result the prominent alterations in histology of the accessory sex organs. Further studies on the physiological and molecular regulation of these androgen-sensitive organs using EDS model will be helpful to understand the normal and pathological development and differentiation mechanism of these organs.

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Expressions of Norepinephrine Transporter in Pre-eclamptic Placenta (자간전증 태반에서의 Norepinephrine Transporter(NET) 발현)

  • Na, Kyu-Hwan;Lee, Hyun-Jung;Jung, Ji-Eun;Kim, Gi-Jin
    • Development and Reproduction
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    • v.14 no.2
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    • pp.65-74
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    • 2010
  • Placenta has been shown to be a site of expression of several of the monoamine membrane uptake transporters. However, the correlation between the expressions of norepinephrine transporter (NET) and placental development including gynecological diseases is still unknown. To investigate the expression and functions of NET in placenta, we conducted to compare NET expression in normal and preeclamptic placenta and analyzed the function of NET in HTR8-SV/neo trophoblast cells after NET gene transfection. The expression of NET was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with preeclamptic placeneta (n=15); and 3) pre-term with preeclamptic placenta (n=11) using semi-quantitative RT-PCR, immunohistochemistry, and Western blot. In order to evaluate the function of NET, NET gene plasmid and NET gene-specific siRNA were trnasfected into HTR-8/SVneo trophoblast cells for 24 hours. NET had low expression in the pre-eclamptic placenta compare with normal placenta but no difference in western blot data. NET was expressed in the trophoblasts, and the up-regulation of NET gene stimulated the invasion of HTR-8/SVneo trophoblast cells by 2.5 fold (p<0.05), whereas the NET-siRNA treatment reduced invasion rates. Also, we observed that the expression of NET induces to expression and activity of MMP-9 in HTR-8/SVneo trophoblast cells in zymography. The results suggest that the expression of NET were reduced in pre-eclampsia and should be inhibited invasion activity of trophoblasts. Therefore, these findings provide useful guidelines for the mechanisms of trophoblast invasion as well as for the basic understanding of gynecological diseases including pre-eclampsia.

Anti-inflammatory Effect of Sargassum coreanum Ethanolic Extract through Suppression of NF-κB Pathway in LPS Induced RAW264.7 Cells in Mice (LPS로 유도된 RAW 264.7 cell로부터 NF-κB 조절 억제와 마우스 모델을 통한 큰잎모자반 에탄올 추출물의 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Bark, Si-Woo;Pak, Won-Min;Ahn, Na-Kyung;Choi, Yeon-Uk;Bae, Nan-Young;Park, Ji-Hye;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.112-119
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    • 2015
  • The anti-inflammatory effect of Sargassum coreanum ethanolic extract (SCEE) was investigated using lipopolysaccharide (LPS)-induced inflammatory responses in this study. It was shown that there was no cytotoxicity in the viability of macrophages treated with SCEE when compared to the control. The production of NO was considerably suppressed by SCEE, approximately up to 50% at 100 μg/ml. This significantly decreased levels of IL-6, TNF-α, and IL-1β. In addition, the expression of iNOS, COX-2, NF-κB was suppressed by SCEE treatment. In in vivo testing, the croton oil-induced mouse ear edema was attenuated by SCEE and there were no mortalities in mice administered with 5000 mg/kg body weight of SCEE over a 2 week observation period. From these results, SCEE inhibits the release of LPS-induced pro-inflammatory cytokines and mediators, suggesting that SCEE could be a potential agent for anti-inflammatory therapies.

Isolation of Synthetic Detergent Decomposing Microorganisms in Wastewater and Synthetic Detergent Decomposition Characterization of the Microorganisms (폐하수중 합성세제분해균의 분리(分離) 및 합성세제 (ABS) 분해특성(分解特性))

  • Lee, Hong-Jae;Heo, Jong-Soo;Cho, Ju-Sik;Han, Mun-Gyu;Choi, Jeong-Ho;Lee, Chun-Hee
    • Korean Journal of Environmental Agriculture
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    • v.12 no.2
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    • pp.144-152
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    • 1993
  • A bacterium which degrades efficiently synthetic detergents was isolated from the polluted waters, activated sludge of wastewater treatment plants or polluted soil. This bacterium showed considerably higher growth rate in the agar plate containing $2,000{\mu}g/ml$ of synthetic detergents than any other isolated strains, was identified as a Pseudomonas fluorescens or strains similar to it. The strain was named as a Pseudomonas fluorescens S1. Optimum pH and temperature for the growth of the Pseudomonas fluorescens S1 were pH 7.0 and $30^{\circ}C$, respectively. The strain was resistant to streptomycin and gentamycin, but sensitive to kanamycin. The strain was greatly resistant to zinc chloride, lead nitrate and copper sulfate, but unable to grow in the presence of relatively low concentrations of mercury chloride and silver nitrate. This strain utilized benzene, catechol, cyclohexane and xylene as a sole carbon source. The strain was well grown in the medium containing ABS 10,000${\mu}g$/ml. Degradation of ABS was 55% and 60% at 20${\mu}g$/ml and 100${\mu}g$/ml of ABS, respectively. Benzene ring was degraded 45% in 100${\mu}g$/ml of ABS. During the incubation of the strain in the medium containing ABS 100${\mu}g$/ml and COD 10,000${\mu}g$/ml for 4 days, degradation of ABS and COD were reduced to 40${\mu}g$/ml and 3,200${\mu}g$/ml, respectively. Total amino acid content of the Pseudomonas fluorescens S1 grown with 1,000${\mu}g$/ml of ABS was 115mg/g cell, whereas its content was decreased in the bacterium grown without synthetic detergent by 9.4%.

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Ultrastructural analysis and quantification of autophagic vacuoles in wild-type and atg5 knockout mouse embryonic fibroblast cells (정상 및 atg5 유전자 제거 섬유아세포에서 자가포식체의 미세구조 및 이들의 정량적 분석)

  • Choi, Suin;Jeon, Pureum;Huh, Yang Hoon;Lee, Jin-A
    • Analytical Science and Technology
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    • v.31 no.5
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    • pp.208-218
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    • 2018
  • Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.

Effects of Exogenous Thyroid Hormone $(T_3)$ on Skeletal Development and Physiological Conditions of Juvenile Black Seabream (Acanthopagrus schlegeli) (감성돔(Acanthopagrus schlegeli) 치어의 골격발달 및 생리적 조건에 미치는 외인성 갑상선호르몬$(T_3)$의 영향)

  • KANG Duk-Young;CHANG Young Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.2
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    • pp.305-312
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    • 1997
  • Juveniles of black seabream, Acanthopagrus schlegeli were fed with the diets containing 0, 10, 20, 50 and 100 ppm of 3,5,3'-triiodo-1-thyronine $(T_3)$ respectively to assess the effect of this hormone on skeletal development and the change of physiological conditions for 50 days. $(T_3)$ treatment lasted for initial 40 days. Fish were fed the prescribed diet by hand to satiation in $2\~4$ times per day. After an initial 40 days period, skeletal development and abnormality were examined, and after a 50 days period, food intake, hepatosomatic index (HSI), thyroid cell height (TCH) and body proximate composition were also examined. Although toed intake was not different among 0, 10 and 20 ppm, the food intake of black seabream fed with the diets containing 50 and 100 ppm of $T_3$was significantly lower than those of 10 ppm. After the initial 40 days of $T_3$ administration, $T_3$ increased the relative growth of operculum, head, caudal fin and pectoral fin to body length, resulting in severe morphological abnormalities at the highest dose. Black seabream treated with 50 and 100 ppm of $T_3$ had abnormal shapes such as lordosis and opercular curl. The HSI parameters were reversely correlation with the dietary concentration of $T_3$. After the initial 40 days of this experiment, atrophy of thyroid gland was observed in fish administered with 50 and 100 ppm of $T_3$. On the 50th day of this experiment, atrophy of thyroid gland was observed only in the group administered with 100 ppm of $T_3$, and no difference was observed on TCH among the rest fours of experimental groups. At the end of the experiment the whore body proximate analyses indicated that there were significant effects of $T_3$ level on moisture, protein, lipid and ash contents.

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