We undertook a cross-sectional study in rural Jehlum and urban Karachi to evaluate the prevalence of vitamin D deficiency in Pakistani pregnant women and neonates and to assess any association of serum 25(OH) vitamin D [25(OH)D] concentration with vitamin D binding protein (Gc) genotypes. Altogether, 390 women and 266 neonates were recruited from urban and rural sites, respectively. Serum 25(OH)D was measured by an immunoassay, while Gc genotypes were identified using polymerase chain reaction followed by restriction fragment length polymorphism or PCR-RFLP. One-way analysis of variance or ANOVA and linear regression were used for statistical analysis. In urban Karachi, 99.5% of women and 97.3% of neonates were vitamin D deficient (< 50 nmol/L), while 89% of women and 82% of neonates were deficient in rural Jehlum. Gc genotypes were not associated with serum 25(OH)D concentrations in both women and their neonates. We conclude that vitamin D deficiency is highly prevalent in Pakistani women and their neonates, and Gc genotypes are not associated with serum 25(OH)D concentrations.

Objective:To investigate the relationship of vitamin D deficiency and risk of AMI in a Pakistani population, and to find out any association between vitamin D binding protein (VDBP) genotypes and risk of AMI in this population.Methods:In a comparative cross-sectional study, 246 patients (age: 20-70 years; 171 males and 75 females) with first AMI were enrolled with informed consent. Similarly, 345 healthy adults (230 males and 115 females) were enrolled as controls. Their fasting serum samples were analyzed for 25 (OH) vitamin D, lipids and other biomarkers using kit methods, while DNA was analyzed for VDBP genotypes using PCR-RFLP based methods. Chi-squared test and logistic regression were used for association of vitamin D deficiency and VDBP genotypes with AMI.Results:Mean serum concentration of 25(OH) vitamin D was significantly lower in AMI patients compared to healthy subjects (p=0.015) and percent vitamin D deficiency was higher in AMI patients compared to healthy subjects (p=0.003). VDBP IF-IF genotype was positively associated with the risk of AMI in subject above 45 years after adjusting for potential confounders [OR = 9.86; 95% CI=1.16 to 83.43].Conclusion:Vitamin D deficiency and VDBP IF-IF genotype are associated with AMI in Pakistani adults.

With so many diverse functions such as transporter of vitamin D metabolites and fatty acids, actin scavenger and macrophage activating factor, Gc must have been one of the most conserved proteins in animal kingdom. Our objective was to investigate the evolution of Gc by analyzing its differences at protein level. Using BLAST (Basic Local Alignment Search Tool) searches, Gc amino acid sequences were analyzed for homology. Clustal W2 and Jalview were used for multiple sequence alignment analysis, phylogenetic tree by PhyML 3.0 while Batch Web CD-Search Tool was used for identification for conserved domains within protein sequences. Gc protein percent identity between human and rabbit was 83%, which decreased to 81% with cow, 78% with mouse, 76% with rat, 51% with chicken, 41% with frog and 28% with zebrafish. Phylogram showed that rat Gc was the most diverged, while chicken Gc was the most conserved protein. Analysis also indicated high homology among mammals (human, rabbit, cow, rat, and mouse). Gc is a highly conserved protein in chicken and zebrafish. However, the distance from ancestral protein gradually increased in amphibian (frog) and mammals (human, rabbit, cow, rat, and mouse). Human Gc and rabbit Gc appear to be recently evolved proteins. There appears to be an interesting evolutionary pattern- chicken Gc has the least distance from the ancestral protein, while rat Gc is the most diverged. There is no vertebrate devoid of Gc which is suggestive of its important role in vitamin D metabolism in vertebrates.

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