• International Journal of Oral Biology
• /
• 제31권1호
• /
• pp.1-6
• /
• 2006

The primary cause of periodontitis is plaque-associated anaerobic gram-negative bacteria. As shown in the patients with defects in the number or function of neutrophils, innate immunity plays an important role in resistance to bacterial infection and periodontitis. Toll-like receptor 4(TLR4) is one of the key receptors that recognize the molecular patterns of microbes and initiate innate immune response. To understand the role of TLR4 in the pathogenesis of periodontitis, we investigated whether Asp299Gly of TLR4 mutation is associated with periodontitis in Korean population. Subjects for this study included 90 healthy subjects and 98 periodontitis patients. The Asp299Gly mutation was screened by PCR-Restriction Fragment Length Polymorphism(RFLP) of genomic DNA from blood cells using a primer that creates a NcoI restriction site only in the mutant allele. The Asp299Gly mutation was not found in all subjects tested. Our results suggest that the Asp299Gly mutation of TLR4 is very rare in a Korean population. Further mutation screening may be required to determine the role of TLR4 in the pathogenesis of periodontitis.

• Parasites, Hosts and Diseases
• /
• 제55권6호
• /
• pp.601-606
• /
• 2017

Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures ($T_ms$) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified $T_ms$ at $85.8^{\circ}C$ and $88.6^{\circ}C$, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using $qPCR-T_ms$ analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.

• Journal of Microbiology and Biotechnology
• /
• 제13권1호
• /
• pp.155-158
• /
• 2003

In early 2002, over 200 people in the city of Pusan. Korea suffered from paratyphoid fever resulting from Salmonella Paratyphi A Infection. Antimicrobial susceptibility tests and Xbal pulsed-field gel electrophoresis (PFCE) were conducted to 54 Salmonella Paratyphi A isolated from humans during the period of 1998 to 2002. Most of the isolates ($83\%$) were only nalidixic acid-resistant and $78\%$ were X 1 PFGE patterns. Also, we measured the MIC of ciprofloxacin and screened gyrA mutation(5) using allele- specific PCR and restriction fragment length polymorphism (AS-PCR-RFLP). The representative 5 isolates in 2002 and 1 isolate in 2000 were $1{\mu}g/ml$ of MIC and had mutation at the 83rd codon in gyrA. These data suggest that the outbreak in the early 2002 might have been due to dissemination of the strain present In 2000. Also, decreased susceptibility to ciprofloxacin was partly due to the mutation at the 83rd codon in gyrA.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권5호
• /
• pp.3205-3211
• /
• 2013

Background: Bladder cancer is the second most incident malignancy among Lebanese men. The purpose of this study was to investigate potential risk factors associated with this observed high incidence. Methods: A case-control study (54 cases and 105 hospital-based controls) was conducted in two major hospitals in Beirut. Cases were randomly selected from patients diagnosed in the period of 2002-2008. Controls were conveniently selected from the same settings. Data were collected using interview questionnaire and blood analysis. Exposure data were collected using a structured face-to-face interview questionnaire. Blood samples were collected to determine N-acetyltransferase1 (NAT1) genotype by PCR-RFLP. Analyses revolved around univariate, bivariate and multivariate logistic regression, along with checks for effect modification. Results: The odds of having bladder cancer among smokers was 1.02 times significantly higher in cases vs. controls. The odds of exposure to occupational diesel or fuel combustion fumes were 4.1 times significantly higher in cases vs controls. The odds of prostate-related morbidity were 5.6 times significantly higher in cases vs controls. Cases and controls showed different clustering patterns of NAT1 alleles. No significant differences between cases and controls were found for consumption of alcohol, coffee, tea, or artificial sweeteners. Conclusions: This is the first case-control study investigating bladder cancer risk factors in the Lebanese context. Results confirmed established risk factors in the literature, particularly smoking and occupational exposure to diesel. The herein observed associations should be used to develop appropriate prevention policies and intervention strategies, in order to control this alarming disease in Lebanon.

• 한국수산과학회지
• /
• 제30권5호
• /
• pp.804-808
• /
• 1997

한국산 참굴의 유전적 특성을 조사하기 위하여 한국의 지역별 참굴을 대상으로 mtDNA 제한효소 절편분석과 클로닝을 수행하였다. 지역별로 각각 20개체의 mtDNA에 대하여 8가지 제한효소를 사용하여 DNA 절편양상을 분석한 결과 서해안산 참굴에서는 개체간 차이가 없는 단일 양상이었으며 남해안산의 경우는 두 가지 양상을 보였으며 차이는 HindIII 절단양상에서만 나타났다. 그 중 소수 개체들에서 나타난 양상은 서해안산 개체들에서의 양상과 동일하여 남해안에 서해안산 참굴이 유입되어 혼재하는 것으로 추정되었다. 한국산 참굴의 mtDNA를 대장균 E. coli HB101에서 클로닝하여 유전적 분석을 용이하도록 하였다. 전체 mtDNA를 제한효소를 사용하여 세부분으로 나누어 pUC19 유전자운반체에 클로닝하였다. 클로닝된 재조합 DNA를 제한효소들로 절단하여 한국산 참굴 mtDNA의 제한효소지도를 작성하였다. 남해안산과 서해안산 참굴 mtDNA에서 HindIII 적단 양상이 다르게 나타남을 확인하였고 이는 남해안산이나 서해안산에서 염기치환의 돌연변이에 의한 것으로 사료되었다.

• Journal of Animal Science and Technology
• /
• 제47권2호
• /
• pp.159-166
• /
• 2005

Porcine myostatin(MS1N) gene plays a key role in the differentiation of myoblast and muscle development. Genetic polymorphism was screened by single stranded conformation polymorphism(SSCP) analysis and subsequent DNA sequencing detected a nucleotide substitution(C2150T) in exon 3 of MSIN gene. Phenotypic association of the polymorphism was tested in a Landrace population and positive effects of the allele T for lean growth traits were found in the population. Even though it is not significant, the pigs have IT and TC genotypes were heavier for the body weight at birth and at twenty weeks of age than those containing genotype. Cc. However, the allele T was significantly associated with higher eye muscle area(P < 0.05). As a result of this study, we suggested that the allele T in exon 3 of MSTN gene comes a significant effect for increasing the eye muscle area without decreasing backfat thickness. This polymorphism did not change the amino acid but Taq I -RFLP matched to SSCP band patterns in exon 3 of MSTN gene, which will be an useful molecular marker for breeding of Landrace pigs.

• 한국균학회지
• /
• 제25권3호통권82호
• /
• pp.161-166
• /
• 1997

국내에서 처음으로 Hypoxylon truncatum을 채집하여 형태 및 유전적 특징을 관찰하였다. 채집된 균주의 형태적 특징은 문헌에 기록된 외국의 기존 균주의 특정과 일치하였으며 각기 다른 두 형태의 stromata를 지니고 있었다. 대상균주로부터 DNA를 추출한 후 ITS 부분에 대한 PCR-RFLPs를 이용하여 분석을 하였다. 그 결과, 총 6종의 제한효소로부터 유래된 절편들 중 대상균주간에는 서로 공통되는 절편이 관찰되지 않았다. 이러한 결과를 볼 때 다른 형태의 stromata를 지니고 있는 균주간에는 상당한 유전적 거리가 있다고 할 수 있다. 그러므로 다른 형태의 stromata를 지니고 있는 H. truncatum 균주는 다른 종일 것이라 사료된다. 또한 H. truncatum은 백목이 버섯의 인공재배에 있어서 아주 중요한 역할을 할 수 있으므로 본 균주를 사용하기 전에, 이 균에 대한 정확한 분류학적 개념 확립은 백목이 버섯 인공재배를 위한 정확한 균주 선발에 큰 도움이 되리라 생각된다.

• 한국수정란이식학회지
• /
• 제22권4호
• /
• pp.257-263
• /
• 2007

This study was performed to investigate the relationship between PSS-HSP70 gene polymorphism and artificial insemination (AI) reproductivity in the pigs. The RFLP polymorphism of PSS and the SSCP polymorphisms of HSP70 K1, K3 and K4 PCR product were detected different patterns. In the experiment for AI of fresh semen, spring and fall season showed higher litter size born of 10.89 head than 10.47 head of summer season. Landrace was showed higher litter size of 9.96 head than that of Duroc and Yorkshire (p<0.05). Stress relating PSS and HSP70 polymorphism of PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showd a highest litter size born of 10.97 head and litter size born alive of 10.69 head than that of the other polymorphisms(p<0.05). In the experiment for AI of frozen semen, effects of season and pig breeds were not showed for litter size born. The stress relating polymorphism of PSS-Carrier, HSP70 K1-BB, K3-BB, K4-AB showed highest litter size born of 11.29 head and litter size born alive of 10.82 head and PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showed the lowest litter size born of 8.48 head and litter size born alive of 7.33 head than that of the other polymorphisms(p<0.05). These results suggest that AI litter size born for the stress of forzen thawed semen may be affected by PSS and HSP70 polymorphism in pigs.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제18권1호
• /
• pp.18-27
• /
• 2009

B. thuringiensis strain LY-99 belonging to subsp. alesti (H3a3c), was isolated from Chinese tobacco warehouse and showed significantly high toxicity to Plutella xylostella. For the identification of the cry1-type genes from B. thuringiensis LY-99, an extended multiplex PCRrestriction fragment length polymorphism (PCRRFLP) method was established by using two pairs of universal primers based on the conserved regions of the cry1-type genes to amplify around 2.4 kb cry1-type gene fragments. Then the DNA fragment was cloned into pGEM-T Easy vector and digested with EcoRI and EcoRV enzymes. Through this method, a known cry1-type gene was successfully identified from the reference strain, B. thuringiensis subsp. alesti. In addition, the RFLP patterns revealed that B. thuringiensis LY-99 included a novel cry1A-type gene in addition to cry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novel cry1A-type gene was designated cry1Ah2 (Genbank accession No DQ269474). An inverse PCR method was used to amplify the flank regions of cry1Ah2 gene. Finally, 3143 bp HindIII fragment from B. thuringiensis LY-99 plasmid DNA including 5' region and partial ORF was amplified, and sequence analysis revealed that cry1Ah2 gene from LY-99 showed 89.31% of maximum sequence similarity with cry1Ac1 crystal protein gene. In addition, the deduced amino acid sequence of Cry1Ah2 protein shared 87.80% of maximum identity with that of Cry1Ac2. This protein therefore belongs to a new class of B. thuringiensis crystal proteins.

• 한국가축번식학회지
• /
• 제23권3호
• /
• pp.213-220
• /
• 1999

산자수는 돼지의 번식능력에 있어 경제적 형질 중 중요한 요인으로 작용하고 있다. 최근 IGF-Ⅰ이 임신유지와 태아의 성장발달에 역할을 하는 것은 보고되어 있으나 산자수에 따른 혈청 IGF-Ⅰ연구는 부족한 상태이다. 그러므로 본 연구는 돼지의 산자수 그룹에 따른 혈청 IGF-I 농도 및 산자수에 관여하는 ER 유전자 다형성 분석을 통해 혈청 IGF-Ⅰ 농도에 대한 조사를 하고자 한다. 혈청은 산자수 높은 그룹과 낮은 그룹의 임신 45일부터 105령일 까지 모아 RIA로 IGF-Ⅰ의 농도를 측정하였으며 측정결과 두 그룹간에 유의적인 차이는 보이지 않았으나 점차적으로 떨어지는 경향을 보여 주었다. 또한 ER 유전자 다형성 분석에 따른 혈청내 IGF-Ⅰ농도는 산자수 낮은 그룹을 나타내는 단편의 IGF-Ⅰ농도가 높게 나타내는 단편에 IGF-Ⅰ농도보다 높게 나타났다. 따라서 본 연구를 통해 돼지의 내분비 물질 중 혈청내 IGF-Ⅰ이 돼지의 산자수와는 관련이 없으나 ER의 발현을 동시에 관찰한다면 돼지 산자수률 예측할 수 있다는 가능성을 제시할 수 있으며, 돼지의 번식기관 중 배란율에 관여하는 난소의 IGF-Ⅰ발현양상 등의 연구가 추후 진행될 필요가 있다고 사료된다.