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To quantify differences in ecNOS expression according to the genotypes of the intron 4 VNTR and the E298A polymorphism, ecNOS and GAPDH mRNA were reverse-transcribed and amplified in the same tube (multiplex RT-PCR).
We measured ecNOS protein concentrations, according to the genotypes of the intron 4 VNTR and the E298A polymorphism, by Western blotting; the results are summarized in Fig.
Department of Medicine, Helsinki University, Finland and co-author of the study commented "Our findings indicate that particular alleles and combinations of specific PTHrP VNTR alleles correlate with variability in peak bone mass in the Finnish male population and raise the possibility that this genotypic approach may serve as an important tool in identifying individuals with a genetic predisposition to low bone mass acquisition at a relatively young age and hence a higher risk for developing osteopenia and osteoporosis in later life.
9) detected an association between homozygosity for the 4a allele in the intron 4 VNTR polymorphism of the ecNOS gene and an increased risk of CAD only in current and ex-smokers in the Australian population.
An opposite but significant trend was observed based on the fact that osteoporotic females tended to have a lower frequency of the 252 bp allele of the PTHrP VNTR as compared to healthy females.
At the same time, a multicenter evaluation of recently developed inexpensive and discriminatory VNTR and CRISPR methods is warranted to see if and how they could complement regional surveillance.
MLST and VNTR have been used by numerous research groups to analyze the diversity of outbreak strains and to investigate the origin of epidemics (6,9-77).
The results of the 19-locus VNTR (15 mycobacterial interspersed repetitive unit-VNTR + additional 4 loci) analysis showed that 7 of the 9 RFLP-clusters were identical according to the 19-locus VNTR (online Appendix Figure).
PCR amplification was performed in a reaction mixture of 10 [micro]L containing 5 uJL of Type-it Multiplex Master Mix (QIAGEN, Hilden, Germany), [approximately equal to] 30 ng of DNA template, and VNTR-specific primers for each of the 4 VNTR loci.
Clustered isolates were further genotyped by using an extended panel of 7 hypervariable VNTR loci (27).
This is supported by VNTR data, as these 2 isolates clustered phylogenetically with one of the samples obtained from an outbreak that occurred at the same polo club 3 months prior (groups 5 and 6, Table).
The highest levels of epidemiologic discrimination of strains of the Mycobacterium tuberculosis complex (MTBC) can be achieved by using multilocus VNTR typing, but these results depend on the number and loci used, particularly for homogenous strain groups such as the Beijing family (3-5).