• 제목/요약/키워드: Enzyme susceptibility

검색결과 112건 처리시간 0.03초

Development of Modified Phenylalanine Ammonia-lyase for the Treatment of Phenylketonuria

  • Kim, Woo-Mi
    • Biomolecules & Therapeutics
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    • 제17권1호
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    • pp.104-110
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    • 2009
  • Phenylketonuria (PKU) is an inherited metabolic disorder caused by mutations in the phenylalanine catabolic enzyme, phenylalanine hydroxylase (PAH). The use of phenylalanine ammonia-lase (PAL) by oral and parenteral routes as a therapeutic drug for PKU has been severely limited due to inactivation by intestinal proteolysis and immune reactions. PEGylation was applied to PAL to reduce the degrees of antigenicity and proteolytic inactivation. Kinetic experiments with native PAL and pegylated PALs were performed, and pH stability, temperature stability, and protease susceptibility were evaluated. Enzyme linked immunosorbent assay (ELISA) was carried out to measure the immune complex between pegylated PALs and antiserum that had been extracted from a PAL-immunized mouse. Pegylated PAL, especially branched pegylated PAL (10 kDa, 1:32), was more active for phenylalanine and more stable in pancreatic proteases than native PAL. Native PAL was optimal at pH 8.5, corresponding to the average pH range of the small intestine; the same finding was noted for pegylated PALs. All linear and branched pegylated PALs had low reactivity with mouse antiserum, especially the 1:16 formulation with linear 5-kDa PEG and the 1:32 formulation with branched 10-kDa PEG. Therefore, we suggest the 1:32 formulation with branched 10-kDa PEG as the most promising formulation for enzyme replacement therapy.

압출조리와 단순 열처리에 의한 쌀전분의 분자량 분포변화와 효소민감성에 관한 연구 (Changes in Molecular Weight Distribution and Enzyme Susceptibility of Rice Starch by Extrusion-cooking and Simple Heat-treatment)

  • 김용범;김지용;이철호
    • 한국식품과학회지
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    • 제25권6호
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    • pp.703-709
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    • 1993
  • 쌀가루의 압출 조리조건과 단순 열처리조건에 따른 쌀전분의 분자량 분포와 ${\alpha}-amylase$ 민감성 변화를 조사하였다. 압출조리는 단일축 압출성형기를 이용하였으며 원료의 수분함량 $17{\sim}29%$, 압출온도 $100{\sim}150^{\circ}C$ 범위에서 처리하였다. 단순 열처리는 원통형 ???L슐에 시료를 밀봉하여 $100{\sim}200^{\circ}C$ 유탕기내에서 일정시간 가열하였다. 이들 가열처리된 시료를 다시 ${\alpha}-amylase$를 처리하여 가수분해속도를 측정하였다. 쌀 전분 원료미분의 수분함량이 23% 이하일 때 압출조리시 수분용해도와 분자량 감소가 크게 일어났으며 이러한 변화는 extruder의 기계적 에너지 투입량(MEI)과 비례하였다. 한편 단순 열처리에서는 원료 수분함량이 높아짐에 따라 가열에 의한 분자량 감소가 감지되었으나 그 변화정도는 압출조리에 비교하여 저조하였다 열처리된 시료를 ${\alpha}-amylase$로 처리하였을 경우 수분함량 23% 이하에서 압출조리된 시료는 효소적 가수분해가 크게 일어나 대부분의 전분입자가 5백만 dalton 이하의 분자량을 가지게 되었으나 고수분 압출물과 단순 열처리에서는 효소처리에 의한 분자량 감소가 크게 나타나지 않았다. 이러한 분자량 변화 현상은 전분용액의 고유점도 변화에 의하여 확인되었다. 이러한 결과는 쌀전분의 저분자화에 의한 수용성 증가 및 효소반응 민감성 증대는 단순 열처리보다는 압출조리에 의하여 크게 일어나며 이것도 extruder 내에서 가열과 동시에 가해지는 강한 압력과 층밀림에 의하여 전분입자의 변형이 크게 일어나는 것을 입증하는 것이다.

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Association study between vitamin D receptor gene polymorphism and chronic periodontitis in Koreans

  • Kim, Seon-Jeong;Jang, Dai-Ho;Kang, Byung-Yong;Kim, Hyun-Hee;Lee, Kang-Oh
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 춘계학술대회
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    • pp.177-177
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    • 2003
  • Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RELP) in the vitamin D receptor gene in 141 periodontically healthy controls and 32 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene were detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There were no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

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GSTM1 and GSTT1 Allele Frequencies among Various Indian and non-Indian Ethnic Groups

  • Senthilkumar, K.P.;Thirumurugan, R.
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6263-6267
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    • 2012
  • Background: Glutathione-S-transferase (GST) is an important phase II xenobiotic compound metabolizing enzyme family, involved in tolerance to a particular drug or susceptibility to a diseasec. This study focused the GSTM1 and T1 null allele frequency in the Gujarat population with a comparison across other Inter- and Intra-Indian ethnic groups to predict variation in the possible susceptible status. Methods: DNA was isolated by a salting out method and GSTM1 and T1 homozygous null genotypes were detected by multiplex polymerase chain reaction in 504 unrelated individuals. The genotype distribution of null alleles was compared with Indian and non Indian ethnics reported earlier in the literature using Fisher's test. Results: The frequencies of the homozygous null genotypes of GSTM1 and GSTT1 were 20% (95%CI 16.7-23.9) and 35.5% (95%CI 31.4-39.9) respectively. GSTM1 null frequency did not deviate from most other Indian ethnic groups but differed from the majority of those of non Indian ethnicity studied. The frequency of homozygous null type of GSTT1 was significantly higher and deviated from all Indian groups and a few of non Indian ethnicity. Conclusions: Gujarat ethnicity, possibly the most susceptible for GSTT1 dependent drug disposition and diseases regarding effects of pollution. Further, the results have implications for GSTT1 dependent drugs used for treatment, a serious problem which needs to be solved by physicians and clinical researchers.

경북지역 애견 번식장에서 분리한 Brucella canis의 생화학적특성 및 PFGE 양상 (Biochemical characterization and PFGE pattern of Brucella canis isolated from kennels in Gyoengbuk province)

  • 김성국;김영환;홍현표;엄현정;장성준;조민희;이양수
    • 한국동물위생학회지
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    • 제30권3호
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    • pp.363-374
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    • 2007
  • A biochemical characterization and antimicrobial drugs susceptibility study was conducted in four breeding kennel which was canine abortion caused by Brucella canis in Gyeongbuk province in 2003-2006. Total of 267 dogs domesticated in the four kennel were examination. Among them, 143 (53.6%) dogs were sero-positive and 25 of blood samples were isolated to Brucella canis. At amplification of 35KDa-BCSP gene using PCR, 711 bp DNA fragment was same visible in 25 isolates and B canis RM6/66. Biochemical characterization of B canis isolated was non-hemolytic, no production of $H_2S$, no fermentation of carbohydrates, catalase-positive, oxidase-positive, indol-negative, hydrolyzation of urea, reduction of nitrate and development of thionin dye medium. Using disk-diffusion method, all of 25 strains tested were found to be highly susceptible to tetracycline, aminoglycoside, quinolone, macrolide antibiotics, rifampin and ampicillin in vitro. Using PFGE with restriction enzyme Smi I, 25 isolates tested were typed to 2 pattern, S1 and S2.

Increasing of Macrophage Migration Inhibitory Factor Expression in Human Patients Infected with Virulent Brucella in Iraq

  • Khudhur, Hasan R.;Menshed, Abbas Ali;Hasan, Ahmed Abbas
    • 한국미생물·생명공학회지
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    • 제48권4호
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    • pp.569-573
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    • 2020
  • Brucellosis is a zoonotic disease caused by Brucella infections and humans usually contract this disease from close contact with infected animals or their products, usually via the ingestion of cheese or crude milk. Macrophage migration inhibitory factor (MIF) and Pro- and anti-inflammatory cytokines play an important role in susceptibility/resistance and the immunopathogenesis of Brucella infection. These cytokines are crucial factors in the initiation and progression of protective immunity against Brucella infection but the role of MIF has not been well studied in the human response to intracellular microbes. This study was designed to investigate the effect of MIF expression on Brucella susceptibility. A total of 85 positive rose Bengal tests and 24 samples from healthy individuals were collected for this study and subjected to polymerase chain reaction assays (PCR) of the bcsp31 diagnostic gene. MIF concentrations were evaluated using Enzyme-Linked immunosorbent assay (ELISA) and the results showed that 46 (54%) of the rose Bengal test samples were positive and 39 (46%) were negative for bcsp31 (p ≤ 0.05) and used as the gold standard for all of the comparisons in this study. The ELISA results indicate that the mean concentration of MIF was significantly higher in patients with positive rose Bengal tests when compared to the control groups and that its concentration increases with increasing age in both the patient and control groups (p ≤ 0.05).

Antimicrobial susceptibility and pathogenic genes of Staphylococcus aureus isolated from the oral cavity of patients with periodontitis

  • Kim, Ga-Yeon;Lee, Chong Heon
    • Journal of Periodontal and Implant Science
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    • 제45권6호
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    • pp.223-228
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    • 2015
  • Purpose: The goal of this study was to characterize the patterns of antimicrobial resistance and virulence genes in samples of Staphylococcus aureus (S. aureus) isolated from periodontitis patients. Methods: From July 2015 to August 2015, oral saliva was collected from a total of 112 patients diagnosed with periodontitis, including 80 outpatients in dental hospitals and 32 patients in dental clinics located in Seoul and Cheonan. The samples were subjected to a susceptibility test to evaluate the prevalence of antimicrobial resistance, and the pathogenic factors and antimicrobial resistance factors in the DNA of S. aureus were analyzed using polymerase chain reaction. Results: A susceptibility test against 15 antimicrobial agents showed that 88% of cultures were resistant to ampicillin, 88% to penicillin, and 2% to oxacillin. Resistance to at least two drugs was observed in 90% of cultures, and the most common pattern of multidrug resistance was to ampicillin and penicillin. Enterotoxins were detected in 65.9% of samples. The cell hemolysin gene hld was detected in 100% of cultures and hla was detected in 97.6% of samples. All strains resistant to penicillin and ampicillin had the blaZ gene. The aph(3')IIIa gene, which encodes an aminoglycoside modifying enzyme, was detected in 46.3% of samples. Conclusions: In the treatment of oral S. aureus infections, it is important to identify the pathogenic genes and the extent of antimicrobial resistance. Furthermore, it is necessary to study patterns of antimicrobial resistance and cross-infection in the context of periodontological specialties in which antimicrobials are frequently used, such as maxillofacial surgery, where the frequency of antimicrobial use for minor procedures such as implant placement is increasing.

녹농균의 Exoenzyme 산생능 및 임상검체별로 본 항균제 감수성 (Production of Exoenzyme of Pseudomonas aeruginosa and Susceptibility to Antimicrobial Agents)

  • 최병주;조양자
    • 대한미생물학회지
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    • 제16권1호
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    • pp.19-28
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    • 1981
  • The Pseudomonas infection has been increased in incidence and suspected as a cause of opportunistic pathogen. Protease and elastase produced by Pseudomonas aeruginosa are reported to be closely associated with pathogenicity of Pseudomonas aeruginosa. We examined, in this work, the relationship between production of exoenzyme of Pseudomonas aeruginosa and susceptibility to antimicrobial agents in view of possible application to the management of Pseudomonas infection. 1. In 295 Pseudomonas aeruginosa isolated from clinical specimens, 34.6% were from pus, 20.7% from sputum, 15.6% from wound including burn sites and 12.9% from urine. 2. Distribution of protease and elastase production by clinically isolated Pseudomonas aeruginosa, showed that protease and elastase producing strains were 83.1%, protease producing strains were 7.5%, elastase producing strains were 2.0%, and non producing strains were 7.5%. 3. MIC(minimum inhibitory concentration) peak for tetracycline and chloramphenicol were observed at 25mcg/ml and 200mcg/ml respectively, but there were no Pseudomonas aeruginosa which correspond to MIC peak, 6.25mcg/ml. Gentamicin of aminoglycosides was highly susceptible to Pseudomonas aeruginosa clinically isolated from pus, sputum and wound sites, but susceptible to isolates from nasal discharge and urine. Regarding MIC peak of carbenicillin, 100mcg/ml, 81.8% of Pseudomonas aeruginosa were from urine, 54.8% from wound including burn sites, 52.7% from pus, and 50.8% from sputum. 4. Enzyme producing strains showed no susceptibility to kanamycine and carbenicillin at low concentration, but protease producing strains tend to resistant to antimicrobial agents.

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한국인 구강 편평세포암에서 Glutathione S-transferase와 CYP1A1 유전자의 다형성 (GENETIC POLYMORPHISMS OF THE GLUTATHIONE S-TRANSFERASE AND CYP1A1 GENES IN KOREAN ORAL SQUAMOUS CELL CARCINOMA)

  • 차인호;권종진;박광균
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제28권5호
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    • pp.364-371
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    • 2002
  • Many chemical compopunds are converted into reactive electrophilic metabolites by the oxidative(Phase I) enzymes, which are mainly cytochrome P-450 enzyme(CYPs). Phase II conjugating enzymes, such as glutathione S-transferase(GST), usually act as inactivation of enzymes. Genetic polymorphisms have been found to be associated with increased susceptibility to cancer of the lung, bladder, breast and colorectal. Many of the polymorphic genes of carcinogen metabolism show considerably different type of cancer among different ethnic groups as well as individuals within the same group. The aim of this study is (1) to establish the frequencies of genetic polymorphisms of GSTM1 and CYP1A1 in Korean oral squamous cell carcinoma(SCC), (2) to associate oral SCC with the risk of these genetic polymorphisms. The genetic polymorphisms of the GSTM1 and the CYP1A1 genes among 50 Korean oral SCC were analyzed using polymerase chain reaction(PCR). The results suggest that the homozygote and the mutant type of CYP1A1 MspI polymorphisms may be associated with genetic susceptibility to oral SCC in Korean. A combination of the GSTM1 null type with the homozygote(m1/m1), and the mutant(m2/m2) type of CYP1A1 MspI polymorphisms showed a relatively high risk of oral SCC in Korean. In the smoking group, the GSTM1 wild genotype may be the high risk factor of oral SCC in Korean. These data coincide with the hypothesis which states that different susceptibility to cancer of genetic polymorphisms exist among different ethnic group and different types of human cancer.

Genome-Wide Association Study of Liver Enzymes in Korean Children

  • Park, Tae-Joon;Hwang, Joo-Yeon;Go, Min Jin;Lee, Hye-Ja;Jang, Han Byul;Choi, Youngshim;Kang, Jae Heon;Park, Kyung Hee;Choi, Min-Gyu;Song, Jihyun;Kim, Bong-Jo;Lee, Jong-Young
    • Genomics & Informatics
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    • 제11권3호
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    • pp.149-154
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    • 2013
  • Liver enzyme elevations, as an indicator of liver function, are widely associated with metabolic diseases. Genome-wide population-based association studies have identified a genetic susceptibility to liver enzyme elevations and their related traits; however, the genetic architecture in childhood remains largely unknown. We performed a genome-wide association study to identify new genetic loci for liver enzyme levels in a Korean childhood cohort (n = 484). We observed three novel loci (rs4949718, rs80311637, and rs596406) that were multiply associated with elevated levels of alanine transaminase and aspartate transaminase. Although there are some limitations, including genetic power, additional replication and functional characterization will support the clarity on the genetic contribution that the ST6GALNAC3, ADAMTS9, and CELF2 genes have in childhood liver function.