• Title/Summary/Keyword: Marker enzyme

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Water Extract from Rice Bran Fermented with Lactobacillus plantarum Hong Inhibits Thromboxane A2 Production Associated Microsomal Enzyme Activity in Human Platelets

  • Kim, Hyun-Hong;Hong, Jeong Hwa;Ingkasupart, Pajaree;Lee, Dong-Ha;Yeo, DaNa;Park, Hwa-Jin
    • Biomedical Science Letters
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    • v.21 no.4
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    • pp.188-197
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    • 2015
  • In this study, we investigated the effect of rice bran water extract fermented with Lactobacillus plantarum Hong (RBLw), on activities of cyclooxygenase-1 (COX-1) and thromboxane $A_2$ synthase (TXAS), thromboxane $A_2$ ($TXA_2$) production associated microsomal enzymes and evaluated its the antiplatelet effect. RBLw, containing 13.5 mg of ferulic acid, dose-dependently inhibited ADP-induced platelet aggregation, and inhibited the production of $TXA_2$, an aggregation molecule. In addition, RBLw directly inhibited COX-1 activity in a dose-dependent manner, but not TXAS activity in platelet microsomal fraction having cytochrome c reductase (an endoplasmic reticulum marker enzyme) activity and expressing COX-1 (72 kDa) and TXAS (60.5 kDa) proteins. These results suggest that RBLw selectively inhibited the activity of COX-1 rather than TXAS to attenuate $TXA_2$ production in ADP-activated platelets. Thus, we demonstrate that RBLw might have direct COX-1 antagonistic function for platelet aggregation-mediated diseases, such as thrombosis, myocardial infarction, atherosclerosis, and ischemic cerebrovascular disease.

Effects of Samiunkyungtang on inflammation and fecal enzymes in ulcerative colitis animal model (궤양성 대장염 동물모델에서 사미연견탕(四味軟堅湯)의 항염증 및 장내세균 효소활성 억제효과)

  • Kim, So-Yeon;Ryu, Bong-Ha;Park, Jae-Woo
    • The Journal of Korean Medicine
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    • v.29 no.3
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    • pp.50-62
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    • 2008
  • Objectives: This study was aimed to find the effect of Samiunkyungtang on inflammation and microflora in an ulcerative colitis animal model. Methods: We established four groups of normal, control, test 1, and test 2 and assigned 6 rats toeach group. The normal group was not treated by any process and fed by normal saline. The control & test groups were provided with 4% dextran sodium sulfate (DSS) treatment for 7 days. Samiunkyungtang extract was orally administered to test groups (test 1=25mg/kg, test 2 100mg/kg) 3 days after DSS treatment for 10 days. After DSS treatment finished, we sacrificed the mice and measured colon length and enzyme activities such as myeloperoxidase (MPO), alkine phosphatase (ALP), ${\beta}$-glucuronidase, ${\beta}$-glucosidase, chondroitinase, and tryptophanase. Results: The colon lengths of test 1 and 2 groups were longer than the control group (p<0.05). Histologically, the crypts and superficial epitheliums of test 1 and 2 groups were regenerated. Goblet cells from all test groups were retrieved. The inflammatory biochemical marker, MPO and ALP activities in all test groups were highly reduced (p<0.01) compared to the control group. The activities of fecal bacterial enzymes in test groups such as ${\beta}$-glucuronidase, ${\beta}$-glucosidase, chondroitinase, and tryptophanase were reduced compared to the control group (p<0.01). Conclusions: As a result of this experiment, Samiunkyungtang is considered to have an inhibitory effect on inflammation and fecal enzyme activity in DSS-induced colitis animal model. Our results indicate that Samiunkyungtang may possess therapeutic effect on the development of DSS-induced colitis.

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Na+/K+-ATPase Alpha Subunit in the Monogonont Rotifer, Brachionus koreanus: Molecular Cloning and Response to Different Salinity

  • Kim, Hokyun;Lim, Bora;Kim, Byung-Do;Lee, Young-Mi
    • Korean Journal of Environmental Biology
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    • v.34 no.2
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    • pp.97-106
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    • 2016
  • $Na^+/K^+$-ATPase is a membrane protein and plays a key role in osmotic regulation in living organisms. In the present study, a cDNA sequence encoding the $Na^+/K^+$-ATPase alpha subunit from the monogonont rotifer, Brachionus koreanus was cloned by rapid amplification of cDNA ends technique. To investigate the role of this enzyme in osmotic stress, enzymatic activities of $Na^+/K^+$-ATPase were measured after exposure to different salinities for 48 h. The full-length Bk $Na^+/K^+$-ATPase cDNA was 3069 bp-long, encoding a 1022-amino acid polypeptide. Bk $Na^+/K^+$-ATPase possesses eight membrane spanning regions and five conserved domains. Phylogenetic analysis showed that Bk $Na^+/K^+$-ATPase had high identity with those of other species, and was closely clustered with other Brachionus sp. These findings indicate that this protein was conserved both structurally and functionally. B. koreanus $Na^+/K^+$-ATPase activity was stimulated in both hyposaline (6 psu) and hypersaline (32 psu) conditions, suggesting that this protein may play a role in osmoregulation. This study would provide better understanding of the physiology of B. koreanus and this enzyme may be useful as a molecular marker for evaluation of osmotic stress in aquatic environment.

Production of Polyclonal Antibody against $\alpha$-Fetoprotein and Polyclonal Antibody-Based Competitive Enzyme-Linked Immunosorbent Assay for $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein (AFP)에 대한 폴리클로날 항체의 생산 및 $\alpha$-fetoprotein 측정용 효소면역분석법 (competitive ELISA)의 개발)

  • Michung Yoon
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.115-123
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    • 1997
  • $\alpha$-Fetoprotein (AFP) has been a useful marker in screening and/or monitoring patients with hepatocellular carcinoma, gonadal germ cell tumor, gastric carcinoma and neural tube defects. In the present study, it was attempted to produce anti-human AFP polyclonal antibodies and to develop a competitive enzyme-linked immunosorbent assay (ELISA) for the measurement of AFP in human plasma and amniotic fluid. AFP was isolated from amniotic fluid using an isolation procedure consisting of affinity chromatography and preparative polyacrylamide gel electrophoresis. The antibody directed against AFP was raised in rabbits. Double immunodiffusion and Western blotting methods showed that the antiserum was highly specific, reacting with only AFP-containing samples. Standard curve was obtained by using purified AFP and specific antiserum. The assay sensitivity was 5ng/ml and the working range was 5~l,000ng/ml. The within-assay and between-assay coefficient of variance (CV) was 4.5% and 8.5%, respectively. These results indicate that the assay is valuable for the measurement of AFP and found to be simple, reproducible, and accurate.

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BRD7 Promoter Hypermethylation as an Indicator of Well Differentiated Oral Squamous Cell Carcinomas

  • Balasubramanian, Anandh;Subramaniam, Ramkumar;Narayanan, Vivek;Annamalai, Thangavelu;Ramanathan, Arvind
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1615-1619
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    • 2015
  • Background: Promoter hypermethylation mediated gene silencing of tumor suppressor genes is considered as most frequent mechanism than genetic aberrations such as mutations in the development of cancers. BRD7 is a single bromodomain containing protein that functions as a subunit of SWI/SNF chromatin-remodeling complex to regulate transcription. It also interacts with the well know tumor suppressor protein p53 to trans-activate genes involved in cell cycle arrest. Loss of expression of BRD7 has been observed in breast cancers and nasopharyngeal carcinomas due to promoter hypermethylation. However, the genetic status of BRD7 in oral squamous cell carcinomas (OSCCs) is not known, although OSCC is one of the most common among all reported cancers in the Indian population. Hence, in the present study we investigated OSCC samples to determine the occurrence of hypermethylation in the promoter region of BRD7 and understand its prevalence. Materials and Methods: Genomic DNA extracted from biopsy tissues of twenty three oral squamous cell carcinomas were digested with methylation sensitive HpaII type2 restriction enzyme that recognizes and cuts unmethylated CCGG motifs. The digested DNA samples were amplified with primers flanking the CCGG motifs in promoter region of BRD7 gene. The PCR amplified products were analyzed by agarose gel electrophoresis along with undigested amplification control. Results: Methylation sensitive enzyme technique identified methylation of BRD7 promoter region seventeen out of twenty three (74%) well differentiated oral squamous cell carcinoma samples. Conclusions: The identification of BRD7 promoter hypermethylation in 74% of well differentiated oral squamous cell carcinomas indicates that the methylation dependent silencing of BRD7 gene is a frequent event in carcinogenesis. To the best of our knowledge, the present study is the first to report the occurrence of BRD7and its high prevalence in oral squamous cell carcinomas.

The Relationship between Lipid Peroxidation of Red Blood Cell Membrane, and Mean Corpuscular Volume and Liver Enzyme Markers in Alcohol Dependence Patients (알코올 의존 환자의 적혈구막 지질과산화 정도와 적혈구 평균 용적 및 간 효소 지표와의 관련성)

  • Jeong, Seong Yun;Choi, Ihn-Geun;Suh, Guk-Hee;Kang, Hee Jung
    • Korean Journal of Biological Psychiatry
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    • v.5 no.2
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    • pp.235-242
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    • 1998
  • Objectives : Alcohol-induced oxidative stress has been known to injure various tissues or organs. This stress is related with free radicals which are produced as the result of long-term alcohol consumption. Malonyldialdehyde(MDA) is produced by the interaction of free radicals and cell membrane lipids, and indicates the degree of lipid peroxidation indirectly. The purpose of this study was to investigate the relationship between red blood cell(RBC) membrane lipid peroxidation by free radicals, and associated hepatic injuries and hematologic changes. Methods : Thirty-three subjects diagnosed as alcohol dependence according to DSM-IV diagnostic criteria were evaluated within 72 hours after discontinuing alcohol drinking. Clinical characteristics were evaluated by CAGE questionnaire and Korean Michigan Alcoholism Screening Test(MAST). RBC membrane MDA level was measured as the marker of RBC membrane lipid peroxidation. Aspartate aminotransferase(AST), alanine aminotransferase(ALT) and gamma-glutamyltransferase(GGT) were used as the biochemical markers of liver damage due to alcohol ingestion. The alcohol-induced hematologic change was assessed by mean corpuscular volume(MCV). Results : The results were as follows. Clinical characteristics were not different between two groups having normal and abnormal levels of AST, ALT, GGT or MCV. The levels of MDA were not correlated with the clinical characteristics and serum levels of AST, ALT and GGT. However, there was a significant correlation between the levels of MDA and the value of MCV(p=0.017). Conclusions : These findings suggest that oxidative stress in alcohol dependence may not be reflected in liver enzyme markers such as AST, ALT and GGT, but may be reflected in MCV.

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Antimutagenic and Quinone Reductase Inducing Activities of Hericium erinaceus Extracts (노루궁뎅이 버섯 추출물의 항돌연변이원성 및 Quinone Reductase유도 효파)

  • 박선희;김옥미;이갑랑
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1287-1292
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    • 2001
  • The effect of Herricium erinaceus on the mutagenicity in salmonella assay and quinone reductase activity in hapalclc7 cells were studied. Antimutagenic as evaluated by Ames test, the extract and fractions of JHerricium erinaceus had no effects on the mutagenicity by themselves. However, methanol extract and fractions from Hericium erinaceus showed strong inhibitory effect on the mutagenesis induced by N-methyl -N'-nitor-N- nitroso-guanidine (MNNG) and benzo(a)pyrene (B(a)P). Among the solvent fractions of methanol extract, the hexane fraction, the chloroform fraction and the ethylacetate fraction exhibited stronger inhibitory activity against MNNG and B(a)P induced mutagenesis than butanol and water fractions. The methanol extract, the extract, the chloroform and the ethylacetate fractions of Hericium erinaceus induced the activity of quinone reductase, an anticarcinogenic marker enzyme, in murine hepalclc7 cells while the others did have little effect on the enzyme activity.

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Molecular Authentication of Scrophularia herbs by PCR-RFLP Based on rpl-5 Region of Mitochondrial DNA (현삼속 식물의 종판별을 위한 Mitochondrial DNA의 염기서열 및 PCR-RFLP 분석)

  • Lee, Jeong-Hoon;Jo, Ick-Hyun;Lee, Jei-Wan;Park, Chun-Geun;Bang, Kyong-Hwan;Kim, Hong-Sig;Park, Chung-Berm
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.173-179
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    • 2010
  • This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.

The relation between serum levels of epidermal growth factor and necrotizing enterocolitis in preterm neonates

  • Ahmed, Heba Mostafa;Kamel, Nsreen Mostafa
    • Clinical and Experimental Pediatrics
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    • v.62 no.8
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    • pp.307-311
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    • 2019
  • Purpose: Necrotizing enterocolitis (NEC) is one of the most serious complications of prematurity. Many risk factors can contribute to the development of NEC. The epidermal growth factor (EGF) plays a major role in intestinal barrier function, increases intestinal enzyme activity, and improves nutrient transport. The aim of this study was to assess the role of epidermal growth factor in the development of NEC in preterm neonates. Methods: In this study, 130 preterm neonates were included and divided into 3 groups, as follows: group 1, 40 preterm neonates with NEC; group 2, 50 preterm neonates with sepsis; and group 3, 40 healthy preterm neonates as controls. The NEC group was then subdivided into medical and surgical NEC subgroups. The serum EGF level was measured using enzyme-linked immunosorbent assay. Results: Serum EGF levels (pg/dL) were significantly lower in the NEC group (median [interquartile range, IQR], 9.6 [2-14]) than in the sepsis (10.1 [8-14]) and control groups (11.2 [8-14], P<0.001), with no significant difference between the sepsis and control groups, and were positively correlated with gestational age (r=0.7, P<0.001). A binary logistic regression test revealed that low EGF levels and gestational ages could significantly predict the development of NEC. The receiver-operating characteristic curve for EGF showed an optimal cutoff value of 8 pg/mL, with 73.3% sensitivity, 98% specificity, and an area under the curve of 0.92. Conclusion: The patients with NEC in this study had significantly lower serum EGF levels (P<0.001), which indicated that EGF could be a reliable marker of NEC in preterm neonates.

The changes of stresses and ecdysteroid biosynthesis gene expression levels in Kynurenine 3-monooxygenase mutant Bombyx mori

  • Jeong, Chan Young;Lee, Chang Hoon;Kim, Su Bae;Kang, Sang Kuk;Ju, Wan-Taek;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Park, Jong Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.43 no.1
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    • pp.29-36
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    • 2021
  • Silkworms have long been bred with human history to produce silk. It has been with humans for longer than other industrial insects, and the silkworm variety has been continuously improved. Silkworms have been developed into the optimal form for producing high quality silk and pupae. Recently, the production of transgenic silkworms has further expanded the possibility of industrial value of silkworms. Kynurenine 3-monooxygenase (KMO), which is a flavin enzyme, is known for its involvement in ommochrome pigment synthesis. In the field of mammals, including humans, previous studies have revealed the function and role of KMO, which is an important enzyme for various immune responses and cell protection. However, in the case of insects, the function of KMO has only been studied to be involved in the formation of pigment, and accordingly, KMO is used exclusively on screening for generation of transgenic insects as a marker. In this study, using KMO-edited silkworms, it was intended to discover the novel functions and roles of KMO in silkworms by identifying changes in the expression of various genes associated with stress and growth. The changes were observed in expressions of genes regulating on stresses to survive and those on ecdysteroid hormone between wild-type (WT) silkworms and kmo mutant silkworms. The loss of KMO, in particular, decreased the expression of the shadow (sad) gene, one of the Halloween genes in the synthesis of ecdysteroid. In conclusion, these results suggest that silkworm KMO is responsible for potential functions regarding stress response and ecdysteroid synthesis.