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Chain Reaction-restriction Fragment Length Polymorphism Method (chain + reaction-restriction_fragment_length_polymorphism_method)

Distribution by Scientific Domains
Medical Sciences80%

Kinds of Chain Reaction-restriction Fragment Length Polymorphism Method

  • polymerase chain reaction-restriction fragment length polymorphism method
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    Selected Abstracts

    Identification of shrimp species in raw and processed food products by means of a polymerase chain reaction-restriction fragment length polymorphism method targeted to cytochrome b mitochondrial sequences

    ELECTROPHORESIS, Issue 15 2008
    Ananías Pascoal
    Abstract A novel PCR-RFLP method has been developed for the identification of six commercially relevant penaeid shrimp species in raw and processed food products. The method can be completed within 8,h. To implement the method, PCR amplification with the crustF/crustR primers, targeted to the amplification of a ca. 181,bp region of the cytochrome b (cytb) mitochondrial gene in penaeid shrimps, was coupled to restriction analysis with CviJI, DdeI and NlaIV. The method was also applied successfully to the identification of shrimp species in complex processed foods, including this type of shellfish as an added-value food ingredient. The small size of this molecular target facilitates amplification from fresh, frozen, or precooked samples, where DNA fragmentation may be relevant and fragment size critical. We also report the first cytb mitochondrial sequences described to date for the species Farfantepenaeus notialis, Parapenaeus longirostris and Pleoticus muelleri, and these nearly triplicate current knowledge of reference nucleotide sequences in this mitochondrial region for this group of species. The cytb mitochondrial gene may also be considered as a molecular marker for identification and phylogenetic purposes in penaeid shrimp species. [source]

    Investigation of matrix metalloproteinase-1 ,1607 1G/2G polymorphism in a Turkish population with periodontitis

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 12 2008
    Kemal Ustun
    Abstract Aim: Matrix metalloproteinase-1 (MMP-1) is a proteolytic enzyme that degrades extracellular matrix and plays a fundamental role during destruction of periodontal tissues. The aim of this study was to examine the association between MMP-1 ,1607 1G/2G polymorphism and chronic periodontitis susceptibility in a Turkish population. Material and Methods: A total of 180 subjects were enrolled in this study. All the subjects received a periodontal examination including full-mouth clinical attachment loss measurements, probing depths, plaque index scores, gingival index scores and radiographic bone loss ratios. Three groups formed according to periodontal conditions were healthy, moderate periodontitis and severe periodontitis groups. MMP-1 ,1607 1G/2G gene promoter polymorphism was genotyped using a polymerase chain reaction-restriction fragment length polymorphism method. Results: Analysis of the polymorphism showed no differences in distribution of the MMP-1 ,1607 1G/2G polymorphism among healthy, moderate periodontitis and severe periodontitis groups (p>0.05). When the groups were further stratified by smoking status, we found no significant differences in genotype distributions, allele frequencies and carriage rates among any groups either (p>0.05). Conclusions: On the basis of the results, no significant association is found for the MMP-1 ,1607 1G/2G polymorphism with susceptibility to periodontitis. Moreover, smoking status did not seem to affect this result. [source]

    TLR2 Arg753Gly, TLR4 Asp299Gly and Thr399Ile gene polymorphisms are not associated with chronic periodontitis in a Turkish population

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 7 2007
    Afig Berdeli
    Abstract Aim: Toll-like receptor (TLR) gene polymorphisms could affect the host's ability to respond to microbial pathogens. In this case,control study, the association of TLR2 and TLR4 gene polymorphisms with chronic periodontitis (CP) was investigated. Materials and Methods: Genomic DNA was obtained from the peripheral blood of 83 patients with CP and 106 periodontally healthy subjects. The TLR2 Arg753Gly, Arg677Trp and TLR4 Asp299Gly, Thr399Ile gene polymorphisms were genotyped by the polymerase chain reaction-restriction fragment length polymorphism method. The data were analysed by a ,2 test, logistic regression analysis and the Mann,Whitney U test. Results: The 753Gln allele was found in 6.1% of the CP patients as compared with 6.6% in the control group (p>0.05). The frequency of the 299Gly and 399Ile allele was 2.4% and 1.8% in CP patients. For the healthy subjects, the frequency was 2.8% for the 299Gly and 2.5% for the 399Ile allele (p>0.05). None of the CP patients or healthy subjects showed homozygosity for the TLR2 and TLR4 alleles. Percentage of sites with bleeding on probing and plaque were significantly higher in 299Gly-positive patients compared with 299Gly-negative patients (p<0.05). Conclusion: These results showed that the TLR2 and TLR4 gene polymorphisms studied are not associated with susceptibility to CP in Turkish patients. [source]

    Effect of lansoprazole and rabeprazole on tacrolimus pharmacokinetics in healthy volunteers with CYP2C19 mutations

    JOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 8 2004
    Fumio Itagaki
    The aim of this study was to investigate the effects of the proton pump inhibitors (PPIs), lansoprazole and rabeprazole, on tacrolimus pharmacokinetics in healthy volunteers with mutations in the cytochrome P450 (CYP) 2C19 gene (CYP2C19). An open-label crossover study was performed with 19 healthy subjects. Tacrolimus (2 mg) was administered orally with and without lansoprazole (30 mg per day for 4 days) or rabeprazole (10 mg per day for 4 days). Blood concentrations of tacrolimus were determined before and 1, 2, 4 and 8 h after dosing. Genotyping for CYP2C19 was conducted by a polymerase chain reaction-restriction fragment length polymorphism method. Coadministration of lansoprazole significantly decreased the oral tacrolimus clearance, resulting in an increase in the area under the blood concentration-time curve (AUC0,8) (control vs with lansoprazole: 29.7 ± 3.5 vs 44.1 ± 5.0 ng h mL,1, P<0.05). Large individual variation was observed in the effects of lansorazole on tacrolimus AUC0,8 owing to CYP2C19 genotype status. The percent change for tacrolimus AUC0,8 in subjects with and without CYP2C19 mutant alleles was 81% and 29%, respectively. Coadministration of rabeprazole also increased the mean AUC0,8 of tacrolimus, but the difference was not statistically significant. These observations suggest that drug interaction between tacrolimus and lansoprazole occurs in subjects with higher lansoprazole blood concentrations corresponding to CYP2C19 genetic status. In contrast, rabeprazole has minimal effect on tacrolimus pharmacokinetics regardless of CYP2C19 genotype status. [source]

    TNF-, and intPLA2 genes' polymorphism in anorexia nervosa

    ACTA NEUROPSYCHIATRICA, Issue 6 2004
    Agnieszka Slopien
    Objective:, The aim of this study was the assessment of ,308G/A tumor necrosis factor (TNF)- , gene polymorphism and intPLA2 gene polymorphism in patients with anorexia nervosa (AN) and healthy controls. Subjects:, We studied 91 non-related patients with AN and 144 healthy women (blood donors and students). The mean age of women from study group was 18.22 years (SD ± 3.13 years) and from control group was 31.71 years (SD ± 8.22). Methods:, ,Gene polymorphisms were studied with the use of polymerase chain reaction-restriction fragment length polymorphism method. TNF-, gene polymorphism consists of G/A substitution in ,308 promoter region. IntPLA2 gene polymorphism is related to intron 1, in which restrictive region is found and recognized by BanI enzyme. Results:, We did not obtain statistically significant differences in the frequency of genotypes and alleles of ,308G/A TNF-, polymorphism between the study and control groups (genotypes: P = 0.106, alleles: P = 0.076). We did analogous analysis in the restrictive and bulimic subgroups. We did not observe statistically relevant differences in the frequency of genotypes (P = 0.700) and alleles (P = 0.305). We did not obtain statistically relevant difference in the frequency of genotypes and alleles of intPLA2 gene between the study group and controls (genotypes: P = 0.300, alleles: P = 0.331). We did analogous analysis in both subgroups of AN. We did not observe statistically relevant differences in the frequency of genotypes (P = 0.344) and alleles (P = 0.230). Conclusions:, There was no statistically relevant trend for the association between TNF-, polymorphism and AN. We did not find association between studied polymorphism of intPLA2 gene and risk of AN. [source]