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MMP-1 and TIMP-1 production in MG-63 cells stimulated with Prevotella nigrescens Lipopolysaccharide (Prevotella nigrescens lipopolysaccharides로 자극된 MG63 세포에서 분비되는 기질금속단백질 MMP-1과 TIMP-1의 수준에 관한 연구)

  • Yang Won-Kyung;Kim Mi-Ri;Shon Won-Jun;Lee In-Bog;Cho Byeong-Hoon;Um Chung-Moon;Son Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.470-478
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    • 2004
  • The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS). and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0, 1, $10{\;}\mu\textrm{g}/ml$) or LPS($10{\;}\mu\textrm{g}/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days. Total RNA was isolated from the cell. and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1. The results were as follows. 1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent. 2. When stimulated with ($1{\;}\mu\textrm{g}/ml$ of LPS. TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However. TIMP-1 expression was suppressed at higher concentration ($10{\;}\mu\textrm{g}/ml$). 3. When P. nigrescens LPS was pretreated with $Ca(OH)_2$. MMP-1 and TIMP-1 gene expression was downregulated. The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS could be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

Association between Genetic Polymorphisms of Blood Proteins and Meat Production Traits in Korean Native Cattle (한우의 혈액단백질의 유전적 다형과 산육형질간의 연관성)

  • Sang, Byung Chan;Han, Sung Wook;Shin, Hyung Doo;Ryoo, Seung Heui;Lee, Sang Hoon
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.68-78
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    • 1998
  • To investigate the genetic polymorphisms and constitutions of blood proteins and enzymes in the Korean native cattle population of National Livestock Cooperatives Federation(NLCF), the genetic variants of transferrin(Tf), post-transferrin-2(pTf-2), albumin(Alb), post-albumin(pAlb), ceruloplasmin(Cp), amylase-I(Am-I) and herroglobin(Hb) were analyzed using the PAGE(polyacrylamide gel electrophoresis) and ST AGE(starch gel electrophoresis) methods. On the genetic variants of the serum proteins, the transferrin(Tf) locus was assumed to be genetically controlled by codominant alleles, Tf A, $D_1$, $D_2$ and E alleles, and the gene frequencies of these were 0.249, 0.248, 0.260 and 0.243, respectively. The post-transferrin locus was observed to be controlled by pTf-2 F and S alleles, and the gene frequencies of these were 0.662 and 0.338, respectively. The post-albumin(pAlb) loci were identified to be controlled by two alleles, pAlb F and S alleles for pAlb locus, and the gene frequncies of these were 0.440 and 0.560 for pAlb F and S alleles, respectively. On the genetic variants of the serum enzymes, ceruloplasmin(Cp) and amylase-I(Am-I) loci were found to be controlled by two alleles, Cp F and S for Cp locus, and Am-I B and C for Am-I locus, and gene frequencies of these were 0.319 and 0.681 for Cp F and S, and 0.871 and 0.120 for Am-I Band C, respectively. On the genetic variants of the hemoglobin(Hb), the distributions of genotypes were 76.5, 21.2 and 2.3% for Hb AA, AB and BB types, and the gene frequnecies for Hb A and B were 0.871 and 0.129, respectively. On the effects of genetic variants of blood proteins, Tf $D_1D_1$, $D_2D_2$ and $D_2E$ genotypes were significantly higher on body weight at 6 month and average daily gain than that of other Tf genotypes.

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Effect of renin inhibition on an experimental glomerulonephritis - a preliminary report (레닌 억제제가 실험적 사구체 콩팥염에 미치는 효과 - 예비 보고)

  • Kang, Ju-Hyung;Heo, Jae-Kyung;Lee, Young-Sook;Han, Ji-Young;Ha, Il-Soo
    • Clinical and Experimental Pediatrics
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    • v.52 no.8
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    • pp.938-943
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    • 2009
  • Purpose : We performed this study in order to investigate the effect of direct renin inhibition on an experimental animal model with nephrotoxic serum nephritis and tried to give useful information for clinical research and renin inhibitor treatment. Methods : Thirty BALB/c 6-week-old male mice were divided into 4 groups: control group (CO, n=5), control-treatment group with aliskiren (CT, n=5), disease group (DO, n=10), and disease treatment group with aliskiren (DT, n=10). Nephritis was induced by an intravenous injection of 0.25 mg/g weight of rabbit anti-GBM immunoglobulin G. Model 2002 Alzet mini-osmotic pumps (Durect Corp.) for aliskiren infusion were implanted into CT and DT. Each group strain was sacrificed serially one at a time on day 14. We estimated the protein-creatinine ratio in 12-hour-collected urine (UP/Cr) and measured the mesangial matrix score in the PAS-stained kidney of each strain. Results : One strain at CT and DT died on day 6 and 7, respectively. Each group strain was sacrificed serially at a time on day 10 because DO were seriously ill. The UP/Cr of each group is as follows: CO, $31.24{\pm}6.54mg/mg$, CT, $23.38{\pm}13.60mg/mg$, DO, $112.72{\pm}10.97mg/mg$, DT $114.07{\pm}32.30mg/mg$. There was no significant difference between DO and DT. The mesangial matrix score of each group was CO, $0.23{\pm}0.10$; CT, $0.13{\pm}0.03$; DO, $1.90{\pm}0.48$; and DT, $1.28{\pm}0.41$, respectively, and there was a significant difference between DO and DT in the extent of mesangial matrix expansion (P=0.008). Conclusion : We found that renin inhibition was able to suppress the mesangial matrix expansion in experimental mice with acute nephritis, although there were no significant differences in UP/Cr.

Renal Effects of a Low Protein Diet and Antihypertensive Drugs on the Progression of Early Chronic Renal Failure in 5/6 Nephrectomized Rats (저단백 식이 및 항고혈압제의 투여가 만성신부전증의 진행에 미치는 영향에 관한 실험적 연구)

  • Kim, Kyo-Sun;Kim, Kee-Hyuk;Kim, Sang-Yun;Kang, Yong-Joo;Maeng, Won-Jae
    • Childhood Kidney Diseases
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    • v.2 no.2
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    • pp.125-132
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    • 1998
  • Purpose : To study whether a low protein diet increase the efficacy of antihypertensive therapy on the progression of renal failure, we conducted an experimental study using 5/6 nephrectomized rats(n=63). Methods : At 7 days after surgery, rats were randomly assigned to three groups according to receiving antihypertensive drug: no antihypertensive drug (U), enalapril (E), and nicardipine (N), respectively and fed a low protein diet (6$\%$ protein). Proteinuria, mesangial matrix expansion score and glomerular volume were assessed at 4, 12 and 16 weeks after renal ablation. Results : Group U rats on a low protein diet developed progressive hypertension ($140{\pm}8,\;162{\pm}5,\;171{\pm}5\;and\;184{\pm}11\;mmHg$ at 4, 8, 12 and 16 weeks) which were controlled by E and N. Group U rats on a low protein diet developed proteinuria ($74{\pm}15\;mg/day$ at 16 weeks) which were decreased by E ($42{\pm}12 mg/day$) or N ($48{\pm}8 mg/day$) (p<0.05). Mesangial matrix expansion score and glomerular volume were not different between groups U, E and N on a low protein diet regardless of the antihypertensive drugs administered. Conclusion : A low protein diet did not affect blood pressure. Enalapril and nicardipine-treated rats on a low protein diet did not have different mesangial matrix expansion and glomerular volumes from rats on a low protein diet at 12 weeks and 16 weeks, in spite of the better controlling of systemic hypertension and lessening of proteinuria. Thus, combined treatment with a low protein diet and antihypertensive drugs didn't appear to show any addition,11 effects to attenuate glomerular injury.

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Diagnosis of Salmonella dubin in Korean Native Calves using PCR and Nucleotide Sequences of rfb5 Gene (송아지에 감염된 Salmonella dublin의 PCR 진단과 rfbS 항원단백 유전자의 염기서열분석)

  • 김철민;이영준;박명규;최경성;김민석
    • Journal of Veterinary Clinics
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    • v.17 no.2
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    • pp.464-469
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    • 2000
  • An epizootic of calf diarrhea occurred in a Korean native cattle farm located in Chonbuk province. The calves that had either bloody or watery diarrhea were 1 to 30 days old. Some of these animals died during the acute course of the disease. Five calves with predominant clinical signs were examined in more detail. Hematological and serum chemical findings were suggestive of dehydration and nutritional insufficiency. Fecal material from the calve was cultured on/in brilliant green agar (BGA), xylose-lysine deoxycholate (XLD) medium, MacConkey agar, eosin methylene blue (EMB) agar and triple sugar iorn (TSI) A bacteria was isolated. which was subsequently identificed as belonging to Salmonella spp. To differentiate Salmoenlla serotype, rfbs gene of S. dublin was amli- find (720 bp) by multiplex (PCR). The rfbS gene sequences of S, dublin ficld isolate(SDC-1) was com- pared with that off S. dublin(S-37) S, dublin(Ahn et al, 1996), S enteritidis(Ahn et al 1996)and S. typhi (Generbak accession No M29682). The identities of nucleotide sequences were 100%. 99.6%, 99.6%, 97.5% respectively.

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Roles of Matrix Metalloproteinases on Intracellular Staphylococcus aureus Growth in Bronchial Epithelial Cell (황색포도알균의 감염에 따른 세포 내에서의 균의 증식과 Matrix Metalloproteinase (MMP)의 역할)

  • Min, Bo Ram;Lee, Young Mi;Park, Jae Seok;Choi, Won-Il;Kwon, Kun Young
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.1
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    • pp.22-27
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    • 2008
  • Background: Staphylococcus aureus frequently colonizes and infects hospitalized patients. Respiratory infections with Staphylococcus aureus are common in patients with compromised airway defenses. However the mechanisms of S. aureus invasion from colonization to the epithelium are unclear. Cell invasion by S. aureus would require destruction of the extracellular matrix, which is believed to be the result of increased matrix metalloproteinases (MMP) activity. Methods: In this study, respiratory epithelial cells were infected with S. aureus. After removing the extracellular bacteria by washing, the internalized bacteria in the cells were assessed by counting the colonized forming units (CFUs). The cell adhesion proteins, dysadherin and E-cadherin, were evaluated by Western blotting. The MMPs in the bacterial invasion were evaluated by pretreating the cells with GM6001, a MMP inhibitor. Results: The internalization of S. aureus was found to be both time and dose dependent, and the increase in MMP 2 and 9 activity was also dependent on the incubation time and the initial amount of bacterial inoculation. The invasion of S. aureus was attenuated by GM6001 after 12 hours incubation with a multiply of infection (MOI)=50. The expression of dysadherin, a membrane protein, was increased in a time and dose dependent manner, while the expression of E-cadherin was decreased. Conclusion: MMPs may mediate the invasion of S. aureus into epithelial cells.

Analysis of General Components, Mineral Contents, and Dietary Fiber Contents of Synurus deltoides (수리취(Synurus deltoids)의 일반성분, 무기질 및 식이섬유에 관한 연구)

  • Park, Min-Hee;Choi, Byoung-Gon;Lim, Sang-Hyun;Kim, Kyung-Hee;Heo, Nam-Ki;Yu, Seong-Hee;Kim, Jong-Dai;Lee, Kwang-Jae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.11
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    • pp.1631-1634
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    • 2011
  • This study was conducted to estimate the general components, minerals, and dietary fiber contents of Synurus deltoides. S. deltoides contained 81.1% moisture content, and the proportions of crude fat, crude protein, crude ash, and crude fiber were 0.3%, 4.2%, 2.6%, and 3.5%, respectively. Potassium (3,249.1 mg) was the most abundant component among the minerals in S. deltoides. In addition, S. deltoides contained many other minerals, e.g. calcium (854.8 mg), phosphorus (60.3 mg), magnesium (344.7 mg), sodium (57.3 mg), zinc (1.7 mg), iron (30.9 mg), copper (0.8 mg), and manganese (5.8 mg). Almost all of the mineral contents of S. deltoides were higher than those of Aster scaber and Ligularia fischeri, except for zinc, copper, and manganese. Total dietary fiber (TDF), insoluble dietary fiber (IDF), and soluble dietary fiber (SDF) contents of S. deltoides were 42.6 g, 37.9 g, and 4.7 g, respectively, and these were also higher than those of A. scaber and L. fischeri used in this study. These results suggest that S. deltoides may be a valuable nutrient source.

PARAMYOSIN OF THE ABALONE, NOTOHALIOTIS DISCUS (전복 Paramyosin의 분리 및 그 성질)

  • PYEUN Jae Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.5 no.1
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    • pp.29-38
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    • 1972
  • The protein composition of abalone muscle was estimated with the following result: on a series of samples analyzed, water-soluble protein, $19\~22\%$, salt-soluble protein, $27\~39\%$: alkali-soluble protein, $20\~26\%$ : and stroma $20\~28\%$ : respectively. It was demonstrated by ultracentrifugal analysis that approximately $65\%$ of the salt-soluble protein is accounted for by paramyosin, $30\%$ by actomyosin, and $5\%$ by myosin, respectively. The ultracentrifugally homogenous paramyosin was prepared by BAILEY's ethanol-dried method. It showed a $S^{\circ}\;_{20,\;{\omega}$ of 3.14s, and was completely salted in with KCl beyond $0.35{\mu}$. The intrinsic viscosity at $25^{\circ}C$ was estimated at 3.1. The paramyosin is rich in several amino acids such as arginine, aspartic acid, glutamic acid, etc., and lacking of both proline and tryptophane, in rough accord with other paramyosins reported. The abalone paramyosin did not show ATPase activity over a pH range of 5 to 9,5 even in the presence of Ca++ or Mg++. So was the case with the paramyosin specimen prepared by BAILEY's wet-extraction method.

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MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide (Prevotella nigrescens lipopolysaccharide로 자극한 치주인대 섬유아세포에서 기질금속단백분해효소와 단백분해효소억제제의 생성 양상에 대한 연구)

  • Yang, Won-Kyung;Lee, Woo-Cheol;Kim, Mi-Ri;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.372-384
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    • 2005
  • The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

Correlation Analysis among Milk Yield, Milk Composition, and Somatic Cell Scores by Definition of Contemporary Group (동기우군의 정의에 따른 유량, 유성분, 체세포 점수간 상관분석)

  • Jung, Woon-Young;Cho, Kwang-Hyun;Choi, Tae-Jeong;Choi, Jae-Kwan;Choi, Ho-Sung;Cho, Ju-Hyun;Choy, Yun-Ho
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.113-121
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    • 2012
  • A total of 150,624 records of Holstein milk production collected from 2005 to 2009 were analyzed to investigate the effects of two different contemporary group definitions, parity and somatic cell score (SCS). The first definition (H BY S) of contemporary group was milking cows and heifers born in the same year and season. And the second thing (H CY S) was milking cow and heifers that delivered calves in the same year and season. Effects of contemporary group, parity and regression effect on SCS from two models were highly significant sources of variation. Coverage of variation ($R^2$) was somewhat higher in models with H BY S as contemporary group. From multivariate models with H BY S, phenotypic correlation coefficients of milk components were estimated high and positive. However, the phenotypic correlation coefficient between milk yield and SCS was -0.09, which was low enough to evidence no correlation between them. Phenotypic correlation between SCS and butter fat or between SCS and protein were also negligible but negative. From multivariate models with H CY S as contemporary group, phenotypic correlation among milk traits and SCS were similar to the estimates from models with H BY S. However, SCS in these models were lowly but negatively correlated with milk yield, milk protein, butter fat or SNF, and the phenotypic correlation coefficients of which were -0.10, -0.08, -0.08, -0.11, respectively.