• Title/Summary/Keyword: 중합효소 연쇄반응

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BIOSENSORS IN AGRICULTURAL AND BIOLOGICAL SYSTEMS (농업 및 생물계에서의 바이오 센서)

  • Delwiche, M.J.;Jenkins, D.M.;Tang, X.;Jackson, E.S.
    • Proceedings of the Korean Society for Agricultural Machinery Conference
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    • 2000.11a
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    • pp.76-100
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    • 2000
  • A brief general discussion of the nature and function of biosensors is presented. While the primary motivator for biosensor development has been the health-care industries, recent research efforts have spread to problems in agriculture and biological production systems. To illustrate some of the research from our laboratory, three example biosensors and their corresponding applications are presented. The first of these is an immunosensor for measurement of the hormone progesterone during milking as a method to improve reproductive management of dairy herds. The second example is an enzyme sensor for measurement of urea in milk as a menas to determine the efficiency of conversion of input protein to milk protein and, thus, improve nutritional management of dairy herds. The third example is a DNA sensor using polymerase chain reaction to detect pathogenic bacteria in the wash water of fresh and minimally processed fruits and vegetables. The potential for application of biosensors in agriculture, agrobiotechnology, food processing, and environmental monitoring has barely been realized.

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Sequencing analysis of the OFC1 gene on the nonsyndromic cleft lip and palate patient in Korean (한국인 비증후군성 구순구개열 환자의 OFC1 유전자의 서열 분석)

  • Kim, Sung-Sik;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.33 no.3 s.98
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    • pp.185-197
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    • 2003
  • This study was performed to identify the characteristics of the OFC1 gene (locus: chromosome 6p24.3) in Korean patients, which is assumed to be the major gene behind the nonsyndromic cleft lip and palate. The sample consisted of 80 subjects: 40 nonsyndromic cleft lip and palate patients (proband, 20 males and females, mean age 14.2 years); and 40 normal adults (20 males and 20 females, mean age 25.6 years). Using PCR-based assay, the OFC1 gene was amplified, sequenced, and then searched for similar protein structures. Results were as follows: 1. The OFC1 gene contains the microsatellite marker 'CA' repeats. The number of the reference 'CA' repeats was 21 times, and formed as TA(CA)11TA(CA)10. But, in Koreans, the number of tandem 'CA' repeats was varied from 17 to 26 except 18, and 'CA' repeats consisted of TA(CA)n. 2. Nine allelic variants were found. Distribution of the OFC1 allele was similar between the patients and control group. 3. There was a replacement of the base 'T' to 'C' after 11 tandem 'CA' repeats in Koreans compared with Weissenbach's report. However, the difference did not seem to be the ORF prediction results between Koreans and Weissenbach's report. 4. The BLAST search results showed the Telomerase reverse transcriptase (TERT) and the Nucleotide binding protein 2 (NBP2) as similar proteins. The TERT was a protein product by the hTERT gene in the locus 5p15.33 (NCBI Genome Annotation; NT023089) The NBP2 was a protein product by the ABCC3 (ATP-binding cassette, sub-family C) gene in the locus 17q22 (NCBI Genome Annotation; NT010783). 5. In the Pedant-Pro database analysis, the predictable protein structure of the OFC1 gene had at least one transmembrane region and one non-globular region.

Discrimination of Hanwoo from Holstein and Mixed Beef by DHPLC (변성 고성능 액체 크로마토그래피를 이용한 한우, 젖소 그리고 혼입육의 구분)

  • Ahn, Young-Chang;Cho, Min-Ho;Seo, Jae-Won;Yoon, Il-Kyu;Jung, Duck-Hyun;Lee, Eun-Young;Nam, Youn-Hyoung;Park, Su-Min;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
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    • v.53 no.6
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    • pp.742-748
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    • 2009
  • In the meat industry, correct breed information in food labeling is required to assure meat quality. Genetic markers provide corroborating evidence to identify breed. We described the development of DNA markers to discriminate between Korean beef cattle (Hanwoo), Holstein, and mixed cow beefs. As most breeds are standardized for coat colour, the melanocortin 1 receptor (MC1R) gene, involved in the regulation of eu/pheomelanins synthesis, has been suggested as marker for breed traceability of products of animal origin. We also designed sex-determining region Y (SRY) gene specific primers for Y chromosome detection. In this study, fragments of MC1R gene and SRY gene were amplified by multiplex-PCR and subjected to digestion by MspA1I restriction endonuclease. Reaction products were analysised by denaturing high performance liquid chromatography (DHPLC). As a result, we identified 6 DHPLC peak types from MC1R gene and SRY gene analysis. DHPLC method showed more sensitive than RFLP method for DNA fragments analysis. Therefore, DHPLC method can apply to identify for Hanwoo, Holstein and mixed beef.

TNF-α stimulated IL-8 and IL-10 expression in monocytes from patients with chronic granulomatous disease (만성육아종질환 환자 단핵구에서 TNF-α 자극에 의한 IL-8과 IL-10의 발현 양상)

  • Shin, Kyung-Sue
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1096-1101
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    • 2008
  • Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

Clinical Features, Molecular Analysis, and Outcome of ERT in Korean Patients with Mucopolysaccharidosis Type VI (국내 6형 뮤코다당증의 임상 양상, 분자유전학적 특징 및 효소치료의 효과에 대한 고찰)

  • Wichajarn, Khunton;Kim, Jinsup;Yang, Aram;Sohn, Young Bae;Lee, Beom Hee;Yoo, Han-Wook;Cho, Sung Yoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.1
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    • pp.24-33
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    • 2016
  • Mucopolysaccharidosis type VI (MPS VI) is a rare disease caused by the mutation of ARSB with prevalence range from 1/5,000 in northeast Brazil to 1/2,057,529 births in Czech Republic. In Asia, there is only one published figure in Taiwan of about 1/833,000 births. The exact prevalence in the Korean population is unknown, but we estimated the incidence of MPS VI is about 0.03/100,000 live births. Enzyme replacement therapy (ERT) with recombinant human Arylsulfatase B (rhASB) is a modality for the treatment of MPS VI that reduces the excretion of urine glycosaminoglycan (GAG) and improves joint motion, pulmonary function, and endurance. We presented the clinical features, molecular analysis and outcome of ERT in three Korean MPS VI patients. All patients had the typical characteristic clinical features of MPS IV. Short stature, dysostosis multiplex, corneal opacity and valvular heart disease were found at first presentation, while restrictive lung disease and carpal tunnel syndrome developed later in all patients. Molecular analysis demonstrated novel missense and nonsense mutation in the patients, including p.Ile 67Ser, p.Gly328Arg, $p.Arg191^*$, p.Asp352Asn, and p.Gly17Asp. After ERT, urine GAG was decreased in all patients. Skeletal involvement, corneal opacity, heart valve abnormalities and pulmonary function were not improved with ERT, but it had a better outcome on regarding joint motion and endurance. One patient underwent allogeneic bone marrow transplantation (BMT) prior to ERT, but their clinical response was not improved much after BMT. This study demonstrates clinical phenotypes and molecular analysis of the severe form of MPS VI in Korean patients.

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Prevalence of Feline Panleukopenia Virus in Stray and Household Cats in Seoul, Korea (한국의 서울에 사는 길 고양이와 집 고양이에서 고양이 범백혈구감소증 바이러스의 유병률)

  • Kim, Seung-Gon;Lee, Kyo-Im;Kim, Ha-Jung;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.30 no.5
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    • pp.333-338
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    • 2013
  • We investigated the prevalence of feline panleukopenia virus (FPV) in stray and household cats in different regions of Seoul, Republic of Korea. Blood samples were collected from a total of 200 cats (100 stray cats and 100 household cats) and examined by polymerase chain reaction (PCR). The overall prevalence of FPV was 2%. Among test-positive cats, 3% (3/100) were stray cats and 1% (1/100) was a household cat. The incidence of FPV was higher in juvenile cats (< 1 year, 1.5%) than in adult cats (> 1-year-old, 0.5%). The FPV-positive rates of healthy infected cats and sick cats were 1.9% (3/156) and 2.2% (1/44), respectively. We found the positive rate of vaccinated and unvaccinated cats to be 1.3% (1/77) and 2.4% (3/123), respectively. Unlike antibody tests, FPV antigen tests detected current infections in stray and household cats. Therefore, these tests can help in disease diagnosis and treatment. To our knowledge, our study is the first to survey the prevalence of FPV in different cat populations across Seoul. We found a high prevalence of FPV infection in stray and juvenile cats. Therefore, proper vaccination and surveillance are important to prevent FPV outbreaks.

Effects of Ipriflavone on bone remodeling in the rat calvarial cell (백서 두개관세포에서 Ipriflavone이 골조직 개조에 미치는 영향)

  • Lee, Yong-Seung;Kim, Young-Jun;Lee, Ki-Heon;Hwang, Hyeon-Shik
    • The korean journal of orthodontics
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    • v.35 no.4 s.111
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    • pp.275-285
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    • 2005
  • Ipriflavone (isoprofoxyisoflavone), a synthetic derivative from soy isoflavone diazein, has been shown to inhibit bone resorption and perhaps stimulate bone formation This study was performed to examine the effects of ipriflavone on the proliferation and bone remodeling in rat calvarial cells in vitro The rat calvarial cells were isolated from fetus aged 20 to 21 days and cultured In BGJb media The graded concentration of ipriflavone $(10^{-9}\;10^{-5}M)$ was administered into cultured cells. When the cell proliferation was estimated through the measurement of MTT assay, there was no increase in cellular proliferation of the rat calvarial cell at any ipriflavone concentration. The cellular activity was evaluated through the formation of mineralized nodules stained by alizarin red. The formation of mineralized nodules significantly increased at concentrations of $10^{-8}M,\;10^{-7}M\;and\;10^{-6}M$ ipriflavone. Reverse transcription-polymerase chain reaction analyses (RT-PCR) were done at 7 and 14 days after culture to detect the expression of Bone Sialoprotein (BSP), Type I Collagen (COL I) and Osteocalcin(OCN) As a result, the expressions of BSP and COL I increased on the 7th day of culture and the expression of OCN increased on the 14th day of culture. These results indicate that ipriflavone facilitates the bone remodeling process bvy promoting rat calvarial cell differentiation aid stimulating mineralization through increased expression of extracellular matrix genes. such as BSP. COL I and OCN.

Detection of Astrovirus Infection from Hospitalized Young Children Feces by Reverse Transcription-Polymerase Chain Reaction (중증 위장관염 소아의 대변검체에서 역전사-중합효소 연쇄반응을 이용한 Astrovirus 검출)

  • Park, Hae-Kyung;Woo, So-Youn;Seoh, Ju-Young;Chong, Young-Hae;Seo, Jeong-Wan
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.453-459
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    • 1999
  • Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and co infection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows:1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 18 children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).

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Kawasaki Disease with Influenza A Virus and Mycoplasma pneumoniae Infections: A Case Report and Review of Literature (인플루엔자 A 및 폐렴미코플라스마 감염과 병발한 가와사끼병 1례)

  • Moon, Hyeok Soo;Huh, Jae Seong;Kim, Mi Kyung;Lambert, Mulakwa Morisho
    • Pediatric Infection and Vaccine
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    • v.23 no.2
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    • pp.149-154
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    • 2016
  • Although an association of Kawasaki disease (KD) with infectious agents has been suggested, none have been proven to cause KD. In this case study, we present a case of KD with concurrent onset of influenza and Mycoplasma pneumoniae (MP) infections. A 27-month-old boy presented with prolonged fever, cough, and rhinorrhea. During the initial testing, influenza A infection was identified, and he was treated with oseltamivir. Despite the antiviral therapy, the fever persisted, and he had cervical lymph node enlargement, bilateral conjunctival injection, fissured red lips, strawberry tongue, and erythematous skin lesions on the Bacillus Calmette-$Gu{\acute{e}}rin$ vaccination site. Thus, the patient was diagnosed with KD and was treated with intravenous immunoglobulin (IVIG). The result of the initial antimycoplasma immunoglobulin M (IgM) antibody testing and was positive, and an increased IgM titer from baseline was found in a repeat test. We reviewed the hypotheses on pathogens known to be associated with KD and the etiology of KD. Based on our findings, we suspect that symptoms of KD and coronary artery lesions can occur from various infections besides those caused by Mycoplasma species and influenza viruses.

Characterization and Frequency of Vancomycin Resistance in Staphylococcus aureus Isolated in Korea (국내에서 분리된 포도상구균의 Vancomycin 내성빈도 및 특성)

  • 박성언;김종배
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.201-208
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    • 2000
  • The vancomycin, one of the family of glycopeptide antibiotics, inhibits the synthesis of bacterial cell wall peptidoglycan and has been widely used against gram-positive bacterial infections, especially for a treatment of methicillin resistant S. aureus infection. However, clinical isolate which was intermediately resistant to vancomycin (Mu50: MIC 8 $\mu\textrm{g}$/ml) was isolated in recent years. In this study we performed vancomycin susceptibility test with the increment method and population analysis with clinical isolates S. aureus. Also we did several kinds of tests with three selected isolates (s129: MIC 7 $\mu\textrm{g}$/ml, s134: MIC 7 $\mu\textrm{g}$/ml, s135: MIC 8 $\mu\textrm{g}$/ml) to find out possible mechanism of vancomycin resistance. As a result, the prevalence of vancomycin resistant S. aureus isolates among S. aureus strains resistant to methicillin was 23.3% (25/107). The vancomycin resistances of isolated strains of S. aureus were between those of Mu5O and Mu3 strains. By PCR analysis, none of the isolates with decreased vancomycin susceptibility contained known vancomycin resistant genes such as vanA, vanB, vanC1, vanC2, and vanH. Major bands of 81 kDa, 58 kDa, 33 kDa, 28 kDa were demonstrable in whole cell lysates by SDS-PAGE from all three isolates as well as reference strains. And especially,45 kDa protein was overproduced in Mu50 strains. Among them increased production of NAD$^{+}$-linked-$_{D}$-lactate dehydrogenase (dnLDH) were detected from one clinical strain (s135) and Mu5O strain. From these data, we suggest that the mechanism of vancomycin resistance in these isolates are distinct from that in enterococci.

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