• Title/Summary/Keyword: Genes

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Diagnosis of Enteropathogens in Children with Acute Gastroenteritis: One Year Prospective Study in a Single Hospital (소아의 급성 위장관염의 원인균 진단: 단일 병원에서 1년간의 전향적 연구)

  • Chang, Ju Young;Choi, Ji Eun;Shin, Sue;Yoon, Jong Hyun
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.1
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    • pp.1-13
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    • 2006
  • Purpose: Acute gastroenteritis in children is one of the frequently encountered diseases with relatively high admission rate. The aim of this study is to determine the isolation trends of common and emerging pathogens in acute gastroenteritis in children over a 12-month period in a community hospital. Methods: The study group included the children who were hospitalized to Seoul National University Boramae Hospital from April, 2003 to March, 2004 or visited outpatient clinic from April, 2003 to July, 2003 with presenting features of acute gastroenteritis. Stool specimens were obtained within 2 days after the visit and examined for the following pathogens: rotavirus, adenovirus, Salmonella, Shigella, Vibrio, pathogenic Escherichia coli (E.coli), Campylobacter and Yersinia species. Viral study was done with commercial kits for antigen detection. Identification of the bacterial pathogens was done by culture using selective media. For pathogenic E.coli, polymerase chain reaction (PCR) was done with the target genes related to the pathogenecity of enterotoxigenic E.coli (ETEC), enteropathogenic E.coli (EPEC) and enterohemorrhagic E.coli (EHEC). Results: The 130 hospitalized children and 28 outpatients were included in this study. The majority of children (>93%) were less than 6 years. Pathogens were isolated in 47% of inpatients and 43% of outpatients, respectively. Rotavirus was the most frequently identified pathogen, accounting for 42.3% of inpatients and 29.6% of outpatients. Nontyphoidal salmonella is the most commonly isolated bacterial pathogen (3.9%) in hospitalized children. Pathogenic E.coli (EPEC, ETEC) was detected in 2.1% (2/97) of inpatients and 25% (3/12) of outpatients. EHEC, adenovirus, Campylobacter, Yersinia and Shigella species were not detected in this study. Conclusion: Rotavirus is the most common enteropathogen in children with acute gastroenteritis. Nontyphoidal salmonella and pathogenic E.coli are important bacterial pathogens. Campylobacter species may not be commonly detected organism in hospitalized children with acute diarrhea.

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.

Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan (한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성)

  • Hong, Seuk-Won;Moon, Ji-Young;Lee, Bok-Kwon;Kim, Yung-Bu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.386-395
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    • 2007
  • A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

Polygenic Association of ACE and ACTN3 Polymorphisms with Korean Power Performance (ACE와 ACTN3의 다중유전형질과 근력운동 경기력간의 관계)

  • Kim, Chul-Hyun
    • Journal of Life Science
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    • v.22 no.3
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    • pp.398-406
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    • 2012
  • This study aimed to examine whether the polygenic profile of ACE ID and ACTN3 R577X polymorphisms is associated with muscle power performance in Korean athletes. For this study, 106 top-class power athletes (top-class group), 158 elite power athletes (elite-class group), and 676 healthy adults (control) aged 18-39 yrs were recruited and their genotypes were analyzed. The top-class group showed higher frequencies of the II genotype and I allele in ACE, as well as higher frequencies of the RR genotype and R allele in ACTN3 (top-class vs. control: 41.4% vs. 32.1% for II genotype, 67.1% vs. 57.7% for I allele, p<0.05; 42.3% vs. 29.0% for RR genotype, 65.3% vs. 54.8% for I allele, p<0.05). In the polygenic profile, the top-class group had significantly higher frequencies of combined-II/ID+RR/RX genotype than the control group (top-class vs. control: 82.9% vs. 66.7% for II/ID+RR/RX, p<0.05), and there was even a sharp increase in total genotype score (TGS) in this group compared to the elite-class and control groups ($66{\pm}0.9$ vs. $58{\pm}1.9$ vs. $56{\pm}2.3$, p<0.05). The combined-II/ID+RR/RX genotype showed the possibility of succussion in the top-class muscle power performance with an odds ratio of 2.3 (CI:1.4-4.1, p<0.05). These results suggested that ACE and ACTN3 need to interact with each other to affect muscle-power performance in an additive form. Furthermore, the polygenic profile of ACE and ACTN3 can predict muscle performance with high success in a homogeneous dominant combined genotype (II/ID+RR/RX). A further study could identify and combine other genes into ACE and ACTN3 for muscle strength.

Development of Agrobacterium-mediated Transformation Method for Domestically Bred Chrysanthemum Cultivar 'Moulinrouge' and Genetic Change of Leaf Morphology Using AtSICKLE Gene (아그로박테리움를 이용한 국내개발 국화품종 '무랑루즈'의 형질전환 기술 및 AtSICKLE 유전자를 이용한 엽형 변화 국화 형질전환체 개발)

  • Kim, Yun-Hye;Park, Hyun-Myung;Jung, Ji-Yong;Kwon, Tack-Min;Jeung, Soon-Jae;Yi, Young-Byung;Kim, Gyung-Tae;Nam, Jae-Sung
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.449-455
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    • 2010
  • 'Moulinrouge' was selected as the best regenerating cultivar among 18 different spray-type chrysanthemum cultivars bred in the Gyeongnam Flowers Breeding Research Institute. When the leaf explants from standard- and spray-type chrysanthemum 'Jinba' and 'Moulinrouge' were incubated on MS basal medium supplemented with $0.5mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA, both 'Jinba' and 'Moulinrouge' induced adventitious shoots that can be regenerated into plantlets. Based on these regeneration conditions, we developed an efficient $Agrobacterium$-mediated chrysanthemum 'Moulinrouge' transformation method by using sequential selection of shoots from low ($10mg{\cdot}L^{-1}$) to high ($30mg{\cdot}L^{-1}$) concentrations of kanamycin after co-cultivation of leaf explants with $Agrobacterium$ for 10 days and induction of shoots. All kanamycin resistant plants investigated with genomic PCR analysis carried the report gene, $AtSICKLE$, in their genome. Although expression levels of the report gene in the transgenic plants investigated with RT-PCR were relatively low because of inefficiency of CaMV 35S promoter in chrysanthemum, transgenic lines expressing $AtSICKLE$ efficiently showed leaf epinasty phenotype. We expect that our results will provide a useful method that can perform a high-throughput investigation of genes isolated and studied well in model plants for molecular breeding of chrysanthemum.

Transcriptional Activation and Repression of Cell Cycle Regulatory Molecules by Trichostatin A (Trichostatin A 처리에 의하 세포주기 조절인자들의 전사활성화 및 불활성화)

  • Baek Jong-Soo;Lee Hee-Kyung;Cho Young-Su;Kim Sung-Young;Park Kwan-Kyu;Chang Young-Chae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.994-1004
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    • 2005
  • The dihydrofolate reductase (dhfr) promoter contains cis-acting element for the transcription factors Spl and E2F. Transcription of dhfr gene shows maximal activity during the Gl/S phase of cell cycle. The member of the Spl transcriptional factor family can act as both negative and positive regulators of gene expression. There was a report that Spl-Rb and E2F4-pl30 complexes cooperate to establish stable repression of dhfr gene expression in CHOC400 cells. Here, we examined the role of HDAC in dhfr, cyclin E, and cyclin A gene regulation using the histone deacetylation inhibitor, trichostatin A (TSA) in U2OS and C33A cells, a Rb-positive human osteosarcoma cell line, and a Rb-negative cervical carcinoma cell line, respectively. When the dhfr promoter constructs were applied in U2OS cells, TSA markedly stimulated over 14-fold of dhfr promoter activity through dhfr-Spl sites by the deletion of an E2F element. In contrast, the deletion of dhfr-Spl binding sites completely abolished promoter stimulation by TSA. The dhfr promoter activity including dhfr-Spl sites increased only 2-fold in C33A cells. Promoter activity containing only dhfr-E2F site did not have much effect by the treatment of TSA in both U2OS and C33A cells. On the other hand, treatment with TSA induced significantly mRNA expression of dhfr and cyclin E, whereas levels of cyclin A decreased in U2OS cells, but had no effect in C33A cells. These results indicate that TSA have contradictory effect, activation of dhfr and cyclin E genes on Gl phase, and down-regulation of cyclin A on G2 phase through transcriptional regulation in U2OS cells.

Detection and Differentiation of Intentional and Unintentional Mixture in Raw Meats Using Real-time PCR (Real-time PCR을 이용한 식육원료의 의도적, 비의도적 혼입 판별법 개발)

  • Kim, Kyu-Heon;Kim, Mi-Ra;Park, Young-Eun;Kim, Yong-Sang;Lee, Ho-Yeon;Park, Yong-Chjun;Kim, Sang Yub;Choi, Jang Duck;Jang, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.340-346
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    • 2014
  • In this study, the detection method was developed using real-time PCR to distinguish 4 species (bovine, porcine, horse, and chicken) of raw meats. The genes for distinction of species about meats targeted at 12S rRNA and 16S rRNA parts in mitochondrial DNA. Probes were designed to have a 5' FAM and a TAMRA at the 3' end. This study is to develop 4 species-specific primer and probes about raw materials and real-time PCR on 10 strains to observe the products of non-specific signal for similar species. As a result, any non-specific signal were not detected among each other. Real-time PCR method was developed for quantitation and identification of intentional and unintentional mixture in ground mixed meat (The difference of $C_T$ value between intentional mixture and 100% meat: $${\leq_-}$$ cycles, The difference of $C_T$ value between unintentional mixture and 100% meat: $${\geq_-}$$ cycles). The detection and differentiation of intentional and unintentional mixture in this study would be applied to food safety management for eradication of adulterated food distribution and protection of consumer's right.

Virulence Profile and Antimicrobial Resistance of Escherichia coli from Flies Captured from Agricultural Environment (농업환경에 서식하는 파리에서 분리된 E. coli의 병원성 유전자 및 항생제 내성 조사)

  • Yun, Bohyun;Jang, Youn Jung;Kim, Yeon Rok;Kim, Hwang-Yong;Kim, Won-Il;Han, Sanghyun;Kim, Se-Ri;Ryu, Jae-Gee;Kim, Hyun Ju
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.147-153
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    • 2017
  • The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.

Mapping and Race Specific Reaction of the Resistance Gene Pi45(t) in Rice (벼 도열병 저항성 유전자 Pi45(t)의 균계 특이적 반응과 고밀도지도 작성)

  • Kim, Dong-Min;Ju, Hong-Guang;Yang, Paul;Han, Seong-Sook;Roh, Jae-Hwan;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.43 no.1
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    • pp.42-49
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    • 2011
  • QTL analysis for blast resistance was carried out using 140 $BC_3F_3$ lines derived from a cross between Ilpum as a recurrent parent and Moroberekan as a donor parent. 140 $BC_3F_3$ lines with the parents were inoculated with nine blast isolates. To identify QTLs for resistance to nine blast isolates, 134 SSR markers showing polymorphisms between the parents were genotyped for the 140 $BC_3F_3$ lines. A total of 17 resistance QTLs to nine isolates were detected on chromosomes 2, 3, 4, 6, 7, 9 and 10. The phenotypic variance explained by each QTL ranged from 8.2% to 26.4%. The Moroberekan alleles contributed the positive effect at these 17 QTL loci. In a previous study, the QTL, Pi45(t) for durable resistance to blast was identified using a sequential planting method. To know the relationship between Pi45(t) and the isolate-specific resistance gene, an $F_2$ population was developed from a cross between Ilpum and an introgression line harboring Pi45(t). $F_3$ lines segregating for the Pi45(t) were inoculated to three isolates. $F_3$ lines from the $F_2$ plants with the Moroberekan segment at the target region showed resistance to two isolates. This result seems to indicate that the Pi45(t) and the isolate-specific resistance gene are tightly linked or the resistance is controlled by the same gene(s). The markers linked to genes controlling blast resistance would be useful in developing blast resistance lines in the breeding program.

The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.