• Title/Summary/Keyword: Marker enzyme

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Isolation of Urease Positive Vibrio parahaemolyticus and Urease Production (Urease를 생산하는 Vibrio parahaemolyticus의 분리 및 urease 생산)

  • 김종숙;김영희
    • Journal of Life Science
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    • v.10 no.1
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    • pp.94-100
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    • 2000
  • Urease is an important microbial enzyme and its production is a marker to predict potential pathogenicity. An unusual halophilic bacterium producing urease was isolated from sea product and identified as Vibrio parahaemolyticus KH410. Its biochemical properties were indole negative, gelatin positive, sodium citrate positive and Kanagawa positive whereas other characteristics were identical as the standard strain except it showed a positive reaction on Christensen's urea agar. V. parahaemolyticus urease production was directly related to urea concentration. The production of urease was noticeable by the addition of 0.2% urea, 0.5% glucose, 2% NaCl in LB broth, and the initial pH of 5.5. The maximum production reached after 6 hr of incubation at 37$^{\circ}C$. However, NiCl2, metal ions, phosphorus did not affect production of urease.

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Effects of Extracts of Monascus-fermented Angelica gigas Nakai in High-fat Diet-induced Obesity in Rats (고지방 식이 유도 비만 흰쥐에서 홍국발효 참당귀 추출물의 항비만 효과)

  • Kim, Young Wan;Kim, Tae Hoon;Sim, So Yeon;Ahn, Hee Young;Park, Kyu Rim;Kim, Jung Wook;Cho, Young Su
    • Journal of Life Science
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    • v.28 no.1
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    • pp.58-67
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    • 2018
  • This study aimed to identify the effects of extracts of fermented Angelica gigas Nakai (A. gigas) with a Monascus purpureus strain on anti-obesity in a high-fat diet (HFD)-induced obesity rat model. Male Sprague-Dawley rats were randomly divided into seven dietary groups (n=8 per group), as follows: a normal diet group (N) and six HFD groups (C: control, HFD and no treatment; AG: HFD +10% A. gigas extracts; FAG2.5: HFD +2.5% fermented A. gigas extracts; FAG5: HFD +5% fermented A. gigas extracts; FAG10: HFD +10% fermented A. gigas extracts; GC: HFD + Garcinia cambogia extracts). FAG-fed rats exhibited effectively lowered rates of increasing body weight and visceral fat accumulation in the HFD-induced obesity model. The activities of several hepatic marker enzymes, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP), were increased with HFD-induced obesity, but levels of these marker enzymes were significantly decreased in FAG-fed rats. The consumption of FAG reduced triglyceride concentrations in serum to normal levels. FAG-fed rats showed effectively increased leptin concentrations in the HFD-induced obesity model. HFD ingestion induced a significant increase in the thiobarbituric acid reactive substances (TBARS) levels, which was decreased in FAG-fed rats. Hematoxylin and eosin staining and Oil Red O staining of the liver showed that the lipid deposits were decreased via FAG feeding. Moreover, hematoxylin and eosin staining of epididymal adipose adipose tissue showed that the adipocyte were decreased by feeding FAG.

Isolation and Culture of Purified Aortic Endothelial Cells Derived from Alpha 1, 3-galactosyltransferase-deficient Pigs (이종 장기이식 및 조직 공학을 위한 Alpha gal 유전자 결손돼지(1, 3-galactosyltransferase-deficient pigs)에서 혈관내피세포(aortic endothelial cells)의 구축)

  • Ock, Sun A;Lim, Malgum;Kim, Yeongji;Ullah, Imran;Shin, Yurianna;Kim, Youngim;Oh, Keon Bong;Hwang, Seongsoo;Hur, Tai-Young;Lee, Seunghoon;Im, Gi-Sun
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.87-94
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    • 2017
  • Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

Serum Talin-1 is a Potential Novel Biomarker for Diagnosis of Hepatocellular Carcinoma in Egyptian Patients

  • Youns, Mahmoud M.;Abdel Wahab, Abdel Hady A.;Hassan, Zeinab A.;Attia, Mohamed S.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3819-3823
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    • 2013
  • Background: Hepatocellular carcinoma (HCC) is a major cause of cancer mortality worldwide. The outcome of HCC depends mainly on its early diagnosis. To date, the performance of traditional biomarkers is unsatisfactory. Talins were firstly identified as cytoplasmic protein partners of integrins but Talin-1 appears to play a crucial role in cancer formation and progression. Our study was conducted to assess the diagnostic value of serum Talin-1 (TLN1) compared to the most feasible traditional biomarker alpha-fetoprotein (AFP) for the diagnosis of HCC. Methods: TLN1 was detected using enzyme linked immunosorbent assay (ELISA) in serum samples from 120 Egyptian subjects including 40 with HCC, 40 with liver cirrhosis (LC) and 40 healthy controls (HC). Results: ROC curve analysis was used to create a predictive model for TLN1 relative to AFP in HCC diagnosis. Serum levels of TLN1 in hepatocellular carcinoma patients were significantly higher compared to the other groups (p<0.0001). The diagnostic accuracy of TLN1 was higher than that of AFP regarding sensitivity, specificity, positive predictive value and negative predictive value in diagnosis of HCC. Conclusions: The present study showed for the first time that Talin-1 (TLN1) is a potential diagnostic marker for HCC, with a higher sensitivity and specificity compared to the traditional biomarker AFP.

Development of an Economic-trait Genetic Marker by Applying Next-generation Sequencing Technologies in a Whole Genome (NGS 기법을 활용한 전장게놈에서의 경제형질 관련 유전자 마커 발굴)

  • Gim, Jeong-An;Kim, Heui-Soo
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1258-1267
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    • 2014
  • Developing economic traits with a high growth rate, robustness, and disease resistance in livestock is an important challenge. RFLP and AFLP are the classical methods used to develop economic traits. Whole-genome-based economic traits have recently been detected with the advent of next-generation sequencing (NGS) technologies. However, NGS technologies are rather costly for use in studies, and RNA-seq, RAD-Seq, RRL, MSG, and GBS have been used to overcome the issue of high costs. In this study, recent NGS-based studies were reviewed, particularly those that focused on minimum costs and maximum effects. Then, we presented further prospects on how to apply for selection of high economic-trait livestock.

COX-2 Expression in Renal Cell Carcinoma and Correlations with Tumor Grade, Stage and Patient Prognosis

  • Tabriz, Hedieh Moradi;Mirzaalizadeh, Marzieh;Gooran, Shahram;Niki, Farzaneh;Jabri, Maryam
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.535-538
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    • 2016
  • Background: Cyclooxygenase 2 (COX-2) is important as an enzyme in the pathway leading to the production of prostaglandin E2 (PGE2) and arachidonic acid. This pathway is known to play a role in inflammation, tumor growth, invasiveness and metastasis, inhibition of apoptosis and angiogenesis. Inhibition of COX-2 has been shown to be a promising antitumor and antiangiogenic strategy in several tumor types, including renal cell carcinoma (RCC). Therefore, we decided to evaluate the immunohistochemical expression of this marker and its association with several clinicopathological characteristics in a series of cases. Materials and Methods: COX-2 expression was examined immunohistochemically in tumor tissues obtained from 96 patients who underwent radical (94 cases) or partial (2 cases) nephrectomy. Correlations between COX-2 expression and clinicopathologic findings including pathologic stage, nuclear grade and other indicator of prognosis were examined. Results: Of 96 tumors, 20.9% were positive for COX-2 expression. A correlation was found between COX-2 expression and tumor histological subtype (P=0.03).The papillary subtype showed maximum expression of this marker (43.8%) and the clear subtype minimum (14.7%). There were also possible links between COX-2 expression and pathologic stage, nuclear grade and nodal involvement but the results were not statistically significant (P=0.8, P= 0.14 and P=0.06, respectively). No correlation was found between COX2 expression and patient age, gender, tumor size, metastasis or survival. Conclusions: In our study, COX-2 expression was correlated with the histological subtype of RCC. Additional research is required to determine the link between COX-2 expression and prognosis and also evaluation of probable effectiveness of COX-2 inhibitor drugs in treatment of RCC patients.

The Studies on Hydrolysis of the Silk Fibroin by Proteolytic Enzyme, Bombyx mori III. Variations of the Fibroin Hydrolysis by Different Silkworm Varieties (견 Fibroin의 효소분해에 관한 연구 (III) -가잠의 원종 및 교잡종견사의 Fibroin 분해율 차이-)

  • 이용우;최병희;송기언;남중희
    • Journal of Sericultural and Entomological Science
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    • v.18 no.2
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    • pp.101-105
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    • 1976
  • The study was carried out to investigate the variations in the fibroin hydrolysis of leading silkworm varieties in Korea, and the relationship between the fibroin hydrolysis of the silkworm varieties with some characteristics and silk morphological property. The results obtained are summarized as follows: 1) The fibroin hydrolyzing ratio of both parent silkworm cocoons was reduced in the F$_1$hybrids. 2) A positive corelation (r=+0.86) was approved between the mid-parent value and the F$_1$hybrid in the fibroin hydrolysis. 3) It was disclosed that the fibroin hydrolyzing ratio of Jam 103${\times}$Jam 104 was the lowest among the leading silkworm F$_1$hybrids. 4) The fibroin hydrolysis was affected by the silk morphological property, showing a positive corelation between the size of cocoon filament and the fibroin hydrolyzing ratio. 5) It was thought that the determination of the fibroin hydrolyzing ratio was useful as a marker for the selection of its related economical characters in silkworm breeding, because the fibroin hydrolysis of parents silkworm cocoons was heritable in their F$_1$hybrids.

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Evaluation of the role of ischemia modified albumin in neonatal hypoxic-ischemic encephalopathy

  • Talat, Mohamed A.;Saleh, Rabab M.;Shehab, Mohammed M.;Khalifa, Naglaa A.;Sakr, Maha Mahmoud Hamed;Elmesalamy, Walaa M.
    • Clinical and Experimental Pediatrics
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    • v.63 no.8
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    • pp.329-334
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    • 2020
  • Background: Birth asphyxia is a leading cause of neonatal mortality. Ischemia-modified albumin (IMA) levels may have a predictive role in the identification and prevention of hypoxic disorders, as they increase in cases of ischemia of the liver, heart, brain, bowel, and kidney. Purpose: This study aimed to assess the value of IMA levels as a diagnostic marker for neonatal hypoxic-ischemic encephalopathy (HIE). Methods: Sixty newborns who fulfilled 3 or more of the clinical and biochemical criteria and developed HIE as defined by Levene staging were included in our study as the asphyxia group. Neonates with congenital malformation, systemic infection, intrauterine growth retardation, low-birth weight, cardiac or hemolytic disease, family history of neurological diseases, congenital or perinatal infections, preeclampsia, diabetes, and renal diseases were excluded from the study. Sixty healthy neonates matched for gestational age and with no maternal history of illness, established respiration at birth, and an Apgar score ≥7 at 1 and 5 minutes were included as the control group. IMA was determined by double-antibody enzyme-linked immunosorbent assay of a cord blood sample collected within 30 minutes after birth. Results: Cord blood IMA levels were higher in asphyxiated newborns than in controls (250.83±36.07 pmol/mL vs. 120.24±38.9 pmol/mL). Comparison of IMA levels by HIE stage revealed a highly significant difference among them (207.3±26.65, 259.28±11.68, 294.99±4.41 pmol/mL for mild, moderate, and severe, respectively). At a cutoff of 197.6 pmol/mL, the sensitivity was 84.5%, specificity was 86%, positive predictive value was 82.8%, negative predictive value was 88.3%, and area under the curve was 0.963 (P<0.001). Conclusion: IMA levels can be a reliable marker for the early diagnosis of neonatal HIE and can be a predictor of injury severity.

Effect of Dexamethasone on the Surface Expression of Marker Molecules and Differentiation of Murine B Cells (덱사메타손이 생쥐 B세포의 세포 표면 인식자와 분화에 미치는 영향)

  • Yeo, Seung-Geun;Cha, Chang-Il;Park, Dong-Choon
    • IMMUNE NETWORK
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    • v.6 no.3
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    • pp.138-144
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    • 2006
  • Background: There are at least two different subsets of B cells, B-1 and B-2. The characteristic features and function of B-2 cells in addition to the effect of steroids on B-2 cells are well-known. Although B-1 cells have different features and functions from B-2 cells, the effect of steroids on B-1 cells is not completely understood. Therefore, this study examined the effects of dexamethasone on peritoneal (or B-1 cells) and splenic B cells (or B-2 cells). Methods: Purified B cells were obtained from the peritoneal fluid and the spleens of mice. The isolated B cells were cultured in a medium and after adding different concentrations of dexamaethasone. The cell survival rate was measured by flow cytometry using propidium iodide. The expression level of the B cell surface marker was analyzed by flow cytometry. During the culture of these cells, immunoglobulin secreted into the culture supernatants was evaluated by an enzyme-linked immunosorbent assay. Results: The survival rate of peritoneal and splenic B cells decreased with increasing dexamethasone concentration. However, the rate of peritofieal B cell apoptosis was lower than that of splenic B cells. CDS and B7.1 expression in peritoneal B cells and CD23 and sIgM expression in splenic B cells after the dexamethasone treatment were reduced. When B cells were treated with dexamethasone, the spontaneous IgM secretion decreased with increasing dexamethasone concentration. Conclusion: Dexamethasone induces apoptosis in peritoneal and splenic B cells. However, peritoneal B cells are less sensitive to dexamethasone. The dexamethasone suppressed expression of the surface markers in peritoneal B cells is different from those in splenic B cells.

Genetics and Breeding for Modified Fatty Acid Profile in Soybean Seed Oil

  • Lee, Jeong-Dong;Bilyeu, Kristin D.;Shannon, James Grover
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.201-210
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    • 2007
  • Soybean [Glycine max(L.) Merr.] oil is versatile and used in many products. Modifying the fatty acid profile would make soy oil more functional in food and other products. The ideal oil with the most end uses would have saturates(palmitic + stearic acids) reduced from 15 to < 7%, oleic acid increased from 23 to > 55%, and linolenic acid reduced from 8 to < 3%. Reduced palmitic acid(16:0) is conditioned by three or more recessive alleles at the Fap locus. QTLs for reduced palmitic acid have mapped to linkage groups(LGs) A1, A2, B2, H, J, and L. Genes at the Fad locus control oleic acid content(18:1). Six QTLs($R^2$=4-25%) for increased 18:1 in N00-3350(50 to 60% 18:1) explained four to 25% of the phenotypic variation. M23, a Japanese mutant line with 40 to 50% 18:1 is controlled by a single recessive gene, ol. A candidate gene for FAD2-1A can be used in marker-assisted breeding for high 18:1 from M23. Low linolenic acid(18:3) is desirable in soy oil to reduce hydrogenation and trans-fat accumulation. Three independent recessive genes affecting omega-3 fatty acid desaturase enzyme activity are responsible for the lower 18:3 content in soybeans. Linolenic acid can be reduced from 8 to about 4, 2, and 1% from copies of one, two, or three genes, respectively. Using a candidate gene approach perfect markers for three microsomal omega-3 desaturase genes have been characterized and can readily be used in for marker assisted selection in breeding for low 18:3.

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