• Title/Summary/Keyword: PCR­RFLP

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Identification of mutans streptococci isolated from dental plaque between the bracket and tooth surface in orthodontic patients (교정환자의 브라켓과 치아 경계부에 존재하는 치면세균막내 mutans streptococci 종 및 생물형의 식별)

  • Kim, Mi-Ae;Yoo, So-Young;Kim, Hwa-Sook;Kook, Joong-Ki;Lim, Sung-Hoon;Yoon, Young-Jooh;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.35 no.1 s.108
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    • pp.51-59
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    • 2005
  • The aim of this study was to compare the species and biotypes of mutans streptococci isolated from dental plaques sampled from the interfaces between the bracket aid tooth surface and smooth tooth surfaces In orthodontic patients. Dental plaque was collected from the interfaces between brackets aid teeth (test group), and from smooth tooth surfaces distant from brackets by more that 2mm (control group). The dental plaque collected by a sterilized curette was transferred into a vial of 1 X PBS. The sample in the vial was vigorously vortexed for 1 min and plated ou mitis-salivarius bacitracin (MSB) agar plate using cotton tips. The agar plates were incubated at $37^[\circ}C$ in a candle jar for 2 days, and again incubated for 1 more day at anambient temperature Individual colonies were cultured in TH broth at $37^[\circ}C\;CO_2$ incubator. The PCR-RFLP based on dextranase gene was performed for the identification of mutans streptococci at the species-level For biotyping of mutans streptococci, biochemical tests were performed There was no significant difference of the species of mutans streptococci isolated from both test and control groups However, the biotypes of the mutans streptococci isolated from test and control groups were different. These results may offer the basic data to verify the relationship between the mutans streptococci biotype and enamel decalcification or dental caries in orthodontic patients with fixed appliances.

MHC-DQB1 Variation and Its Association with Resistance or Susceptibility to Cystic Echinococcosis in Chinese Merino Sheep

  • Hui, Wenqiao;Shen, Hong;Jiang, Song;Jia, Bin
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.12
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    • pp.1660-1666
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    • 2012
  • Cystic echinococcosis (CE), one of the world's most geographically widespread diseases, still represents a considerable economic and public health significance, although a variety of methods has been used to control the disease. It has been demonstrated that genetic factors, especially variations in MHC loci, can influence the outcome of CE infection in the human population. The study described here was designed to determine whether variation in MHC-DQB1 was associated with susceptibility or resistance to CE in sheep. If so, it would lay a theoretical foundation for breeding disease resistance sheep in future. This study was carried out on 204 Chinese Merino sheep, including 101 CE sheep and 103 healthy controls. The polymorphism of MHC-DQB1 exon 2 was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, and $x^2$ test was used to compare genotype frequencies between CE sheep and healthy controls. A total of 22 alleles and 42 genotypes were identified in DQB1 exon 2 in Chinese Merino sheep. In addition, $x^2$ test showed that frequencies of DQB1-TaqIaa and DQB1-HaeIIInn genotypes were significantly higher in the healthy group (82.5% and 57.3%, respectively) than that in the CE group (57.4% and 28.9%, respectively) (both p values = 0, OR = 0.286, 0.303, respectively), suggesting that these genotypes appeared to be associated with resistance to CE. Whereas, frequencies of DQB1-TaqIab and DQB1-HaeIIImn genotypes were significantly higher in the CE group (36.9% and 32.0%, respectively), as compared with the healthy group (16.5% and 11.15%, respectively) (p = 0.001, 0.001 and OR = 2.963, 3.629, respectively), indicating that these genotypes might be associated with susceptibility to CE. It is concluded that the genetic polymorphism within MHC-DQB1 might influence immune responses to pathogens, thus leading to the development of CE or protection against CE in Chinese Merino sheep, which would pave the way for breeding disease resistance sheep in future.

Autosomal-dominant Polycystic Kidney Disease in a Family of Scottish Fold Cats (스코티쉬 폴드 고양이 가족에 발생한 상염색체 우성 다낭성 신병)

  • Seo, Kyoung-Won;Kim, Sae-Um;Ahn, Jin-Ok;Coh, Ye-Rin;Han, Sung-Young;Youn, Hwa-Young
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.726-728
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    • 2010
  • Autosomal-dominant polycystic kidney disease (AD-PKD) is common in Persian and Persian-related breeds, and is sporadically reported in Scottish Fold cats. A 5-year-old male Scottish Fold cat was diagnosed with polycystic kidney disease based on screening tests and abdominal ultrasonography and died 3.5 months after diagnosis. The cat had 14 kittens with three queens, including his female sibling, with an age range of 3 months to 8 years. Genetic testing to confirm the genetic transmission of AD-PKD which detects the mutated PKD1 gene was performed. Abdominal ultrasonography confirmed the presence of renal cysts. Nineteen cats were screened in the present study (13 males and 6 females), with an age range of 3 months to 8 years. The results of renal ultrasonography agreed with the genetic test results in the 19 cats in which both tests were performed and 8 cats were diagnosed as ADPKD based on these tests. AD-PKD has not been investigated in cats in South Korea. Moreover, this is the first report of AD-PKD in a family unit of Scottish Fold cats.

Single Nucleotide Polymorphism Exploring the 5'-Regions of Estrogen Receptor-${\alpha}$ Gene and Association With Reproduction Performance and Milk Yield in Hanwoo and Holstein Dairy Cattle (Estrogen Receptor-${\alpha}$ 유전자 5' 영역의 Single Nucleotide Polymorphism의 탐색과 한우와 Holstein에서 번식능력 및 산유능력과의 관계)

  • Yeom, Gyu-Tae;Jeon, Hyang-A;Park, Hae-Geum;Kim, Young Sin;Kim, Hyun;Kim, Jae Hwan;Seong, Hwan-Hoo;Cho, Young Moo;Cho, Jae-Hyeon;Ko, Yeoung-Gyu
    • Reproductive and Developmental Biology
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    • v.38 no.3
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    • pp.123-127
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    • 2014
  • This study was conducted for SNPs in the 5'-regions of estrogen receptor-${\alpha}$ (ESR-${\alpha}$), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genetic improvement was incurred low reproduction performance. The objective of this study was to investigate connections between single nucleotide polymorphisms (SNP) of Estrogen receptor-${\alpha}$ (ESR-${\alpha}$) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to genotype the cattle. The result showed two variants of the ESR-${\alpha}$ gene. The Bgl I cut the 492 bp amplification product into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction fragments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in these breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was $382.18{\pm}10.03$ days, and G/G was $381.69{\pm}5.22$ days. The SPC of A/G was $1.62{\pm}0.16$, and G/G was $1.32{\pm}0.04$. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was $7,253.00{\pm}936.00kg$ and of G/G was $8,747.51{\pm}204.88kg$, showing no significant difference.

Effects of Alcohol Intake, Genotypes of Aldehyde Dehydrogenase 2 and N-Acetyltransferase 2 on the Development of Laryngeal Cancer in Koreans (한국인의 후두암 발생에서 음주, Aldehyde Dehydrogenase 2(ALDH2)와 N-Acetyltransferase 2(NAT2) 유전자 다형성의 역할)

  • Kwon Soon-Uk;Shim Yoon-Sang;Lee Yong-Sik;Hong Seong-Chool;Kim Kwang-Il;Hong Young-Joon;Hong Seok-Il;Kim Hyun-Joo;Kim Heon;Lee Guk-Haeng
    • Korean Journal of Head & Neck Oncology
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    • v.17 no.2
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    • pp.131-138
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    • 2001
  • Objectives: Alcohol intake has been reported to be a risk factor of laryngeal cancer. Since the aldehyde dehydrogenase 2 (ALDH2) genotype is a major determinant of personal alcohol drinking habit, there is a possibility that ALDH2 genotype would be a risk factor for laryngeal cancer. N-Acetyltransferase 2 (NAT2) is a detoxifying enzyme and its polymorphism has been reported to be related to the risk of many environmental cancers. However, studies on the associations between these two genotypes and laryngeal cancer risk are scarce. We have assessed the effects of alcohol intake and the genotype of ALDH2 and NAT2 on the risk of laryngeal cancer in Koreans. Materials and Methods: Eighty-four pathologically proven laryngeal cancer patients and 168 age matched controls were included as the study subjects. Information about alcohol intake and smoking habit was collected using a self administered questionnaire. ALDH2 and NAT2 genotypes were analyzed using PCR-RFLP methods. Results: Alcohol intake was significant as a risk factor for laryngeal cancer (OR : 2.58, 95% CI : 1.24, 5.36), especially for supraglottic laryngeal cancer (OR : 3.24, 95% CI : 1.02, 10.31). Personal drinking habit was closely related with personal smoking habit, which was a potent risk factor of laryngeal cancer. In a stratified analysis according to the level of cumulative smoking amount, drinking was significant neither in light smokers (equal or less than 30 pack-years) nor in heavy smoker (over 30 pack-years). The ALDH2 genotype was significantly associated with the risk of laryngeal cancer in a univariate analysis. The statistical significance, however, disappeared after adjusting alcohol intake using a multiple conditional logistic model. The NAT2 genotype was not significant as a risk factor for laryngeal cancer. Conclusion: Alcohol drinking and ALDH2 genotype would have indirect effects on laryngeal cancer by their correlations with cigarette smoking or with alcohol drinking. It is less likely that the NAT2 genotype would be a potent risk factor of laryngeal cancer.

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Association between HSP70 Genotypes and Oocytes Development on In vitro Maturation/Fertilization in Pig

  • Wee, M.S.;Park, C.K.;Cho, S.R.;Lee, S.S.;Yeon, S.H.;Kim, C.D.;Cho, C.Y.;Choi, S.H.;Sang, B.D.;Son, D.S.;Li, Z.D.;Jin, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1404-1410
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    • 2008
  • This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

No Association of Hypoxia Inducible Factor-1α Gene Polymorphisms with Breast Cancer in North-West Indians

  • Sharma, Sarika;Kapahi, Ruhi;Sambyal, Vasudha;Guleria, Kamlesh;Manjari, Mridu;Sudan, Meena;Uppal, Manjit Singh;Singh, Neeti Rajan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9973-9978
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    • 2014
  • Background: Hypoxia inducible factor-1 alpha (HIF-$1{\alpha}$) is the key regulator of cellular responses to hypoxia and plays a central role in tumour growth. Presence of Single nucleotide polymorphisms (SNPs) in the critical regulatory domains of HIF-$1{\alpha}$ may result in the overexpression of the protein and subsequent changes in the expression of the downstream target genes. The aim of study was to investigate the association of three SNPs (g.C111A, g.C1772T and g.G1790A) of HIF-$1{\alpha}$ with the risk of breast cancer in North Indian sporadic breast cancer patients. Materials and Methods: A total of 400 subjects, including 200 healthy controls and 200 patients with breast cancer were recruited in this study. Genotypes were determined using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) method. Results: The CC and CA genotype frequency of HIF-$1{\alpha}$ g.C111A polymorphism was 100 vs 99% and 0 vs 1% in breast cancer patients and healthy controls respectively. The frequencies of CC, CT and TT genotype of g.C1772T polymorphism were 76 vs 74.5%, 19 vs 21% and 5 vs 4.5% in breast cancer patients and control individuals respectively. There was no significant difference in genotype and allele frequencies of HIF-$1{\alpha}$ g.C1772T polymorphism between cases and control individuals (p>0.05). For g.G1790A genotypes, all patients and controls had only GG genotype. Conclusions: The three HIF-$1{\alpha}$ polymorphisms (g.C111A, g.C1772T and g.G1790A) are not associated with breast cancer risk in North-West Indian patients.

Distribution and Haplotype Associations of XPD Lys751Gln, XRCC1 Arg280His and XRCC1 Arg399Gln Polymorphisms with Nasopharyngeal Carcinoma in the Malaysian Population

  • Visuvanathan, Shaneeta;Chong, Pei-Pei;Yap, Yoke-Yeow;Lim, Chin-Chye;Tan, Meng-Kuan;Lye, Munn-Sann
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2747-2751
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    • 2014
  • Background: DNA repair pathways play a crucial role in maintaining the human genome. Previous studies associated DNA repair gene polymorphisms (XPD Lys751Gln, XRCC1 Arg280His and XRCC1 Arg399Gln) with nasopharyngeal carcinoma. These non-synonymous polymorphisms may alter DNA repair capacity and thus increase or decrease susceptibility. The present study aimed to determine the genotype distribution of XPD codon 751, XRCC1 codon 280 and codon 399 polymorphisms and haplotype associations among NPC cases and controls in the Malaysian population. Materials and Methods: We selected 157 NPC cases and 136 controls from two hospitals in Kuala Lumpur, Malaysia for this study. The polymorphisms studied were genotyped by PCR-RFLP assay and allele and genotype frequenci es, haplotype and linkage disequilibrium were determined using SNPstat software. Results: For the XPD Lys751Gln polymorphism, the frequency of the Lys allele was higher in cases than in controls (94.5% versus 85.0%). For the XRCC1 Arg280His polymorphism, the frequency of Arg allele was 90.0% and 89.0% in cases and controls, respectively and for XRCC1 Arg399Gln the frequency of the Arg allele was 72.0% and 72.8% in cases and controls respectively. All three polymorphisms were in linkage disequilibrium. The odds ratio from haplotype analysis for these three polymorphisms and their association with NPC was 1.93 (95%CI: 0.90-4.16) for haplotype CGC vs AGC allele combinations. The global haplotypte association with NPC gave a p-value of 0.054. Conclusions: Our study provides an estimate of allele and genotype frequencies of XRCC1Arg280His, XRCC1 Arg399Gln and XPD Lys751Gln polymorphisms in the Malaysian population and showed no association with nasopharyngeal cancer.

IL-6-6331 (T/C, rs10499563) is Associated with Decreased Risk of Gastric Cancer in Northern Chinese

  • Yang, Li;Sun, Ming-Jun;Liu, Jing-Wei;Xu, Qian;Yuan, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7467-7472
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    • 2013
  • Background: Polymorphisms of genes encoding cytokines could be potential biomarkers to predict risk of gastric cancer (GC). Here, we investigated the association between the IL-6 -6331 (T/C, rs10499563) polymorphism in its promoter region and GC risk. Methods: In this case-control study of 215 GC cases and 518 non-cancer controls, the IL-6 -6331 (T/C, rs10499563) polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: Individuals with the TC or CC genotype were associated with a significantly decreased risk of GC (OR=0.710, 95%CI: 0.504-0.999, P=0.049) compared with TT wild-type carriers. Ther C allele was also associated with significantly decreased risk of GC (OR=0.715, 95%CI: 0.536-0.954, P=0.023) compared with the T allele. In the stratification analysis, TC or CC genotypes were associated with significantly decreased GC risk in subgroups of males, people older than 60, and H. pylori-positive cases. However, no significant interaction was observed for TC or CC genotypes with H. pylori infection. On stratification with the Lauren classification, TC or CC genotypes were associated with significantly decreased risk of diffuse-type GC (OR=0.497, 95%CI: 0.266-0.925, P=0.027), also in subgroups of males, people older than 60, and H. pylori-positive cases. Conclusions: The IL-6 -6331 (T/C, rs10499563) polymorphism is associated with genetic susceptibility of GC and may have the potential to predict GC risk.

MMP2 Gene-735 C/T and MMP9 gene -1562 C/T Polymorphisms in JAK2V617F Positive Myeloproliferative Disorders

  • Sag, Sebnem Ozemri;Gorukmez, Ozlem;Ture, Mehmet;Gorukmez, Orhan;Topak, Ali;Sahinturk, Serdar;Ocakoglu, Gokhan;Gulten, Tuna;Ali, Ridvan;Yakut, Tahsin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.443-449
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    • 2015
  • Background: Myeloproliferative disorders (MPDs) are clonal hematologic malignancies originating at the level of the pluripotent hematopoietic stem cell. Matrix metalloproteases (MMPs) are proteolytic enzymes that contribute to all stages of malignancy progression. Genetic variants in the MMP genes may influence the biological function of these enzymes and change their role in carcinogenesis and progression. To our knowledge, this is the first investigation of associations between the -735 C/T and -1562 C/T polymorphisms in the MMP2 and MMP9 genes, respectively, and the risk of essential thrombocytosis (ET), and polycythemia vera (PV). Materials and Methods: The case-control study included JAK2V617F mutation positive 102 ET and PV patients and 111 controls. Polymorphisms were determined by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and electrophoresis. Results: No statistically significant differences were detected between patient (ET+PV) and control groups regarding genotype distribution for MMP2 gene-735 C/T and MMP9 gene -1562 C/T polymorphisms and C/T allele frequency (p>0.050). Statistically borderline significance was observed between PV and control groups regarding genotype distribution for the MMP9 gene -1562 C/T polymorphism (p=0.050, OR=2.26, 95%Cl=0.99-5.16). Conclusions: Consequently this study supported that CC genotype of MMP9 gene -1562 C/T polymorphism may be related with PV even if with borderline significance.