• Title/Summary/Keyword: gIII gene

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Association of the CYP1B1 Gene Polymorphism with the Risk of Advanced Endometriosis in Korean Women (한국 여성에서 중증자궁내막증과 CYP1B1 유전자 다형성과의 관련성에 관한 연구)

  • Cho, Yeon Jean;Hur, Sung-Eun;Lee, Ji Young;Song, In Ok;Koong, Mi Kyoung;Moon, Hye Sung;Chung, Hye-Won
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.2
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    • pp.85-95
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    • 2006
  • Objective: To investigate whether polymorphisms of gene encoding CYP1B1 is associated with the risk of endometriosis in Korean women. Methods: We investigated 199 patients with histopathologically confirmed endometriosis rAFS stage III/IV and 183 control group women who were surgically proven to have no endometriosis. The genetic distribution of four different CYP1B1 polymorphisms at $G^{119}-T$, $G^{432}-C$, $T^{449}-C$, and $A^{453}-G$ were analyzed by polymerase chain reaction(PCR) and restriction fragment length polymorphism of PCR products. Results: We found no overall association between each individual CYP1B1 genotype and the risk of endometriosis. The odds ratio of genotype GG/GC+GG/TC+TT/AA compared to GG/CC/CC/AA(reference) was calculated as 2.06 with a 95% confidence interval of 1.003~4.216. Conclusion: This results suggest that CYP1B1 genetic polymorphism may be associated with development of endometriosis in Korean women.

Gynostemma pentaphyllum extract and Gypenoside L enhance skeletal muscle differentiation and mitochondrial metabolism by activating the PGC-1α pathway in C2C12 myotubes

  • Kim, Yoon Hee;Jung, Jae In;Jeon, Young Eun;Kim, So Mi;Oh, Tae Kyu;Lee, Jaesun;Moon, Joo Myung;Kim, Tae Young;Kim, Eun Ji
    • Nutrition Research and Practice
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    • v.16 no.1
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    • pp.14-32
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    • 2022
  • BACKGROUND/OBJECTIVES: Peroxisome proliferator-activated receptor-gamma co-activator-1α (PGC-1α) has a central role in regulating muscle differentiation and mitochondrial metabolism. PGC-1α stimulates muscle growth and muscle fiber remodeling, concomitantly regulating lactate and lipid metabolism and promoting oxidative metabolism. Gynostemma pentaphyllum (Thumb.) has been widely employed as a traditional herbal medicine and possesses antioxidant, anti-obesity, anti-inflammatory, hypolipemic, hypoglycemic, and anticancer properties. We investigated whether G. pentaphyllum extract (GPE) and its active compound, gypenoside L (GL), affect muscle differentiation and mitochondrial metabolism via activation of the PGC-1α pathway in murine C2C12 myoblast cells. MATERIALS/METHODS: C2C12 cells were treated with GPE and GL, and quantitative reverse transcription polymerase chain reaction and western blot were used to analyze the mRNA and protein expression levels. Myh1 was determined using immunocytochemistry. Mitochondrial reactive oxygen species generation was measured using the 2'7'-dichlorofluorescein diacetate assay. RESULTS: GPE and GL promoted the differentiation of myoblasts into myotubes and elevated mRNA and protein expression levels of Myh1 (type IIx). GPE and GL also significantly increased the mRNA expression levels of the PGC-1α gene (Ppargc1a), lactate metabolism-regulatory genes (Esrra and Mct1), adipocyte-browning gene fibronectin type III domain-containing 5 gene (Fndc5), glycogen synthase gene (Gys), and lipid metabolism gene carnitine palmitoyltransferase 1b gene (Cpt1b). Moreover, GPE and GL induced the phosphorylation of AMP-activated protein kinase, p38, sirtuin1, and deacetylated PGC-1α. We also observed that treatment with GPE and GL significantly stimulated the expression of genes associated with the anti-oxidative stress response, such as Ucp2, Ucp3, Nrf2, and Sod2. CONCLUSIONS: The results indicated that GPE and GL enhance exercise performance by promoting myotube differentiation and mitochondrial metabolism through the upregulation of PGC-1α in C2C12 skeletal muscle.

Human extracellular superoxide dismutase (EC-SOD) expression in transgenic chicken

  • Byun, Sung June;Ji, Mi-Ran;Jang, Ye-Jin;Hwang, A-In;Chung, Hee Kyoung;Kim, Jeom Sun;Kim, Kyung-Woon;Chung, Hak-Jae;Yang, Byoung-Chul;Jeon, Iksoo;Park, Jin-Ki;Yoo, Jae Gyu;Kim, Tae-Yoon
    • BMB Reports
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    • v.46 no.8
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    • pp.404-409
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    • 2013
  • Extracellular superoxide dismutase (EC-SOD) is a metallo-protein and functions as an antioxidant enzyme. In this study, we used lentiviral vectors to generate transgenic chickens that express the human EC-SOD gene. The recombinant lentiviruses were injected into the subgerminal cavity of freshly laid eggs. Subsequently, the embryos were incubated to hatch using phases II and III of the surrogate shell ex vivo culture system. Of 158 injected embryos, 16 chicks (G0) hatched and were screened for the hEC-SOD by PCR. Only 1 chick was identified as a transgenic bird containing the transgene in its germline. This founder (G0) bird was mated with wild-type hens to produce transgenic progeny, and 2 transgenic chicks (G1) were produced. In the generated transgenic hens (G2), the hEC-SOD protein was expressed in the egg white and showed antioxidant activity. These results highlight the potential of the chicken for production of biologically active proteins in egg white.

Cellular Iron Uptake from Aqueous Solutions depending on Reaction Conditions by genetically engineered Saccharomyces cerevisiae (재조합 Saccharomyces cerevisiae에 있어서 반응조건에 따른 수용성 철의 생체 흡수)

  • Kim Sang-Jun;Chang Yu-Jung;Park Chung-Ung;Jeong Yong-Seob;Kim Kyung-Suk
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.441-445
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    • 2004
  • Cellular iron uptake was performed in the yeast Saccharomyces cerevisiae that transformed with human ferritin H- and L-chain genes. The recombinant yeasts were enriched in YEP medium supplemented with $2\%$ galactose for 3 days and the iron uptake was followed by incubating the cells with iron in 20 mM MOPS buffer (pH 6.5). The reactions were examined under different conditions including the iron compounds of Fe(II) and Fe(III), the concentration of iron, the concentration of cells and the reaction time. From our results, the recombinant yeast YGH2 producing H-chain ferritin showed higher cellular iron concentration at the cell concentration of 100 mg/ml than 200 mg/ml. Iron presented as Fe(II) rather than Fe(III) was taken up more efficiently. Iron uptake increased slightly when iron was added up to 14.3 mM Fe(II) and then its cellular iron concentration was $16.7{\pm}0.7\;{\mu}mol/g$ cell wet wt. In addition, the iron uptake reaction reached to maximum at about 2 hr incubation.

Monitoring of Environmental Arsenic by Cultures of the Photosynthetic Bacterial Sensor Illuminated with a Near-Infrared Light Emitting Diode Array

  • Maeda, Isamu;Sakurai, Hirokazu;Yoshida, Kazuyuki;Siddiki, Mohammad Shohel Rana;Shimizu, Tokuo;Fukami, Motohiro;Ueda, Shunsaku
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1306-1311
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    • 2011
  • Recombinant Rhodopseudomonas palustris, harboring the carotenoid-metabolizing gene crtI (CrtIBS), and whose color changes from greenish yellow to red in response to inorganic As(III), was cultured in transparent microplate wells illuminated with a light emitting diode (LED) array. The cells were seen to grow better under near-infrared light, when compared with cells illuminated with blue or green LEDs. The absorbance ratio of 525 to 425 nm after cultivation for 24 h, which reflects red carotenoid accumulation, increased with an increase in As(III) concentrations. The detection limit of cultures illuminated with near-infrared LED was 5 ${\mu}g$/l, which was equivalent to that of cultures in test tubes illuminated with an incandescent lamp. A near-infrared LED array, in combination with a microplate, enabled the simultaneous handling of multiple cultures, including CrtIBS and a control strain, for normalization by the illumination of those with equal photon flux densities. Thus, the introduction of a near-infrared LED array to the assay is advantageous for the monitoring of arsenic in natural water samples that may contain a number of unknown factors and, therefore, need normalization of the reporter event.

Genetic Relatedness within Streptococcus pneumoniae Serotype 19F and 23F Isolates in Korea by Pulsed-Field Gel Electrophoresis

  • Lee, Kwang-Jun;Bae, Song-Mee;Hwang, Kyu-Jam;Lee, Young-Hee;Kim, Ki-Sang
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.1-6
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    • 2003
  • The genetic relatedness of multidrug-resistant pneumococcal isolates of serotypes 19F and 23F was investigated. The DNA fragments digested with Sma I were resolved by pulsed-field gel electrophoresis (PFGE). PFGE analysis of 365. pneumoniae isolates showed 13 different patterns. Among 22 isolates of serotype 19F, 9 different PFGE patterns were present and 14 isolates of serotype 23F isolates represented 5 distinct PFGE patterns. Two isolates of serotype 19F and six isolates of serotype 23F shared the same PFGE pattern (Pattern I). Based on the genetic relatedness within the strains (one genetic cluster was defined as having more than 85% homology), we divided the pneumococcal strains into genefic clusters (Ⅰ, II, III, IV, V, and VI). The 22 strains of serotype 19F belonged to five distinct genetic clusters (I, II, III, IV, V and VI) and 14 strains of serotype 23F represented two genetic clusters (I and II ). These results showed that strains of serotype 19F are genetically more diverse than those of serotype 23F, Serotype 19F isolates with PFGE patterns H and I appeared to be less related to those of the remaining PFCE patterns (A to G) (less than 60% genetic relatedness), but those strains were genetically closely related with serotype 23f. These results suggest that the latter isolates originated from horizontal transfer of the capsular type 19F gene locus to 23F pneumococcal genotypes. In conclusion, the multidrug-resistant pneumococcal isolates of serotype 19f and 23F isolated in Korea are the result of the spread of a limited number of resistant clones.

Loss of FHIT Expression in Non-Small Cell Lung Cancer; The Clinical Significance and Effects on Apoptosis and Cell Proliferation Cycle (비소세포 폐암에서 FHIT 유전자의 발현소실의 임상적의의 및 세포고사 및 세포분열주기에 미치는 영향)

  • Kim, Hak-Ryul;Yang, Sei-Hoon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.6
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    • pp.610-620
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    • 2003
  • Background : 3p deletion has been shown to be the most frequently occurring change in lung cancers, suggesting the presence of a tumor suppressor gene in this region. Recent attention has focused on a candidate 3p14.2 tumor suppressor gene, FHIT. Therefore, the association of the expression of FHIT, with apoptosis, cell proliferation cycle and the clinicopathological features, including survival, were investigated Materials and Methods : 83 patients with non-small cell lung cancer, who underwent curative operation, between Jan. 1996 and Aug. 2000, at the Wonkwang university hospital, were analyzed. The expression of the FHIT was identified by immunohistochemical staining, and rate of apoptosis and cell proliferation cycle by flow cytometry. Results : 43% (36/83) of patients exhibited no FHIT expression. The rates of FHIT loss were 52% (28/54), 22% (5/23), 50% (3/6); 30% (11/37), 48% (16/33), 69% (9/13); 54% (30/56) and 22% (6/27), in squamous cell cancers, adenocarcinomas, large cell cancers, TNM stages I, II and III, smokers and non-smokers, respectively. All the differences in FHIT loss rates, according to the histopathology, TNM stages and smoking habits, were statistically significant. The median survival time and 2-year survival rate of the FHIT(-) group were 24 months and 44%, and those of the FHIT(+) group were 25 months and 51% (p>0.05), respectively. The apoptotic rate of the FHIT(-) and FHIT(+) groups were 50.72 (${\pm}13.93$) and 59.38 (${\pm}14.33$)%, respectively (p=0.01). The S- and G1-phase fractions of the FHIT(-) and FHIT(+) groups were 13.93 (${\pm}7.35$) and $51.50({\pm}23.15$)% and 15.65(${\pm}6.59$) and 54.16 (${\pm}20.25$)%, respectively (p>0.05). Conclusion : The loss of FHIT expression was increased to a greater extent with advancing TNM stage, smoking habits and squamous cell cancer compared to the adenocarcinomas. However, no survival differences were found according to the expression of FHIT. The apoptotic rate of the FHIT(+) group was greater than in the FHIT(-) group, but differences in the S- and G1-phase fractions, according to the expression of the FHIT, were not found.

Individual-breed Assignment Analysis in Swine Populations by Using Microsatellite Markers

  • Fan, B.;Chen, Y.Z.;Moran, C.;Zhao, S.H;Liu, B.;Yu, M.;Zhu, M.J.;Xiong, T.A.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1529-1534
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    • 2005
  • Individual-breed assignments were implemented in six swine populations using twenty six microsatellites recommended by the Food and Agriculture Organization and the International Society for Animal Genetics (FAO-ISAG). Most microsatellites exhibited high polymorphisms as shown by the number of alleles and the polymorphism information content. The assignment accuracy per locus obtained by using the Bayesian method ranged from 33.33% (CGA) to 68.47% (S0068), and the accumulated assignment accuracy of the top ten loci combination added up to 96.40%. The assignment power of microsatellites based on the Bayesian method had positive correlations with the number of alleles and the gene differential coefficient ($G_{st}$) per locus, while it has no relationship to genetic heterozygosity, polymorphism information content per locus and the exclusion probabilities under case II and case III. The percentage of corrected assignment was highest for the Bayesian method, followed by the gene frequency and distancebased methods. The assignment efficiency of microsatellites rose with increase in the number of loci used, and it can reach 98% when using a ten-locus combination. This indicated that such a set of ten microsatellites is sufficient for breed verification purposes.

Homologous Expression and Quantitative Analysis of T3SS-Dependent Secretion of TAP-Tagged XoAvrBs2 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract

  • Kim, S.H.;Lee, S.E.;Hong, M.K.;Song, N.H.;Yoon, B.;Viet, P.T.;Ahn, Y.J.;Lee, B.M.;Jung, J.W.;Kim, K.P.;Han, Y.S.;Kim, J.G.;Kang, L.W.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.679-685
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    • 2011
  • Xanthomonas oryzae pv. oryzae (Xoo) produces a putative effector, XoAvrBs2. We expressed XoAvrBs2 homologously in Xoo with a TAP-tag at the C-terminus to enable quantitative analysis of protein expression and secretion. Addition of rice leaf extracts from both Xoo-sensitive and Xoo-resistant rice cultivars to the Xoo cells induced expression of the XoAvrBs2 gene at the transcriptional and translational levels, and also stimulated a remarkable amount of XoAvrBs2 secretion into the medium. In a T3SS-defective Xoo mutant strain, secretion of the TAPtagged XoAvrBs2 was blocked. Thus, we elucidated the transcriptional and translational expressions of the XoAvrBs2 gene in Xoo was induced in vitro by the interaction with rice and the induced secretion of XoAvrBs2 was T3SSdependent. It is the first report to measure the homologous expression and secretion of XoAvrBs2 in vitro by rice leaf extract. Our system for the quantitative analysis of effector protein expression and secretion could be generally used for the study of host-pathogen interactions.

Biodiversity and Phylogenetic Analysis of Streptomyces Collected from Bamboo Forest Soil (대나무 산림토양으로부터 수집한 Streptomyces 속 방선균의 계통학적 다양성)

  • Lee, Hyo-Jin;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.262-269
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    • 2010
  • To investigate a quantitative evaluation of the actinobacteria, we have collected samples from various kinds of bamboo forest soil. Each different layers contained $2.7{\times}10^6-2.7{\times}10^8$ CFU/g of actinobacteria which was the highest in litter layers of Sasa boreali forest soil. We obtained 330 actinobacteria from different layers of bamboo forest soil; litter (100 strains), humus (70 strains), and rhizosphere soil (160 strains). Based on the colony morphology (aerial mycelium, substrate mycelium, and soluble pigment), isolates were divided into thirty-six groups and we selected 50 representative isolates. 16S rRNA gene sequence analysis showed Streptomyces was major actinobacteria (94%) and they were categorized as cluster I (2 strains), II (35 strains), III (6 strains), and IV (7 strains), respectively. The diversity index of 50 Streptomyces collected from the bamboo forest soil was calculated with the Shannon-Wiener method. Bamboo litter showed higher diversity index level of 3.33 than that of humus and rhizosphere soil. Also, antibiotic activities of our isolates were investigated against Botrytis cinerea, Xanthomonas campestris, Xanthomonas axonopodis pv. vesicatoria, and Bacillus cereus and found in 74, 16, 25, and 24 strains, respectively.