Melatonin and Human Cardiovascular Disease

The possible therapeutic role of melatonin in the pathophysiology of coronary artery disorder (CAD) is increasingly being recognized. In humans, exogenous melatonin has been shown to decrease nocturnal hypertension, improve systolic and diastolic blood pressure, reduce the pulsatility index in the internal carotid artery, decrease platelet aggregation, and reduce serum catecholamine levels. Low circulating levels of melatonin are reported in individuals with CAD, arterial hypertension, and congestive heart failure. This review assesses current literature on the cardiovascular effects of melatonin in humans. It can be concluded that melatonin deserves to be considered in clinical trials evaluating novel therapeutic interventions for cardiovascular disorders.Fil: Pandi Perumal, Seithikurippu R.. King Saud University; Arabia SauditaFil: BaHammam, Ahmed S.. King Saud University; Arabia SauditaFil: Ojike, Nwakile I.. King Saud University; Arabia SauditaFil: Akinseye, Oluwaseun A.. University of New York; Estados UnidosFil: Kendzerska, Tetyana. Sunnybrook Health Sciences Center; CanadáFil: Buttoo, Kenneth. Sleep Disorders Center; CanadáFil: Dhandapany, Perundurai S.. Oregon Health And Science University; Estados UnidosFil: Brown, Gregory M.. University of Toronto; CanadáFil: Cardinali, Daniel Pedro. Pontificia Universidad Católica Argentina ; Argentin

Targeted Genome-Wide Enrichment of Functional Regions

Only a small fraction of large genomes such as that of the human contains the functional regions such as the exons, promoters, and polyA sites. A platform technique for selective enrichment of functional genomic regions will enable several next-generation sequencing applications that include the discovery of causal mutations for disease and drug response. Here, we describe a powerful platform technique, termed “functional genomic fingerprinting” (FGF), for the multiplexed genomewide isolation and analysis of targeted regions such as the exome, promoterome, or exon splice enhancers. The technique employs a fixed part of a uniquely designed Fixed-Randomized primer, while the randomized part contains all the possible sequence permutations. The Fixed-Randomized primers bind with full sequence complementarity at multiple sites where the fixed sequence (such as the splice signals) occurs within the genome, and multiplex amplify many regions bounded by the fixed sequences (e.g., exons). Notably, validation of this technique using cardiac myosin binding protein-C (MYBPC3) gene as an example strongly supports the application and efficacy of this method. Further, assisted by genomewide computational analyses of such sequences, the FGF technique may provide a unique platform for high-throughput sample production and analysis of targeted genomic regions by the next-generation sequencing techniques, with powerful applications in discovering disease and drug response genes

Details of 2092 random population samples from India used to study the 5 bp Deletion Polymorphism.

<p>I/D Insertion/Deletion, D/D Deletion/Deletion, I/I Insertion/Insertion.</p

The hn-RNP's and SR-proteins binding site sequences in controls and DCM as predicted by “Splicing Rainbow” tool.

<p>The hn-RNP's and SR-proteins binding site sequences in controls and DCM as predicted by “Splicing Rainbow” tool.</p

Chi-square and Fisher Exact Probability Test for SNP's found in this study.

<p>*SNP- single nucleotide polymorphism, DCM- Dilated cardiomyopathy.</p

The amino acid arginine at residue 144 in human Troponin T (<i>cTnT</i>) is highly conserved across many species, including mouse, rat, chicken, rabbit, sheep and bovine.

<p>The amino acid arginine at residue 144 in human Troponin T (<i>cTnT</i>) is highly conserved across many species, including mouse, rat, chicken, rabbit, sheep and bovine.</p

Clinical details of the family members carrying R144W mutation.

<p>SCD- Sudden cardiac death; NYHA-New York Heart Association; LVIDd- left ventricular internal diastolic dimension; LVEF- left ventricular ejection fraction.</p

Total number of mutations observed in Troponin T (<i>cTnT</i>) gene.

<p>*SNP- single nucleotide polymorphism, AA-Amino Acid, CON- Controls, DCM- Dilated cardiomyopathy, SS- Splice Site, HP-Highly Polymorphic.</p

1A-1M: Electropherograms showing SNPs of cTnT gene, observed in the present study on South Indian dilated cardiomyopathy patients.

<p>Mutation sites were shown with arrows. <b>Fig. 1A</b><b>. R144W</b> [rs483352832]: Electropherogram (arrow) showing a novel missense mutation (R144W) at the nucleotide position g.14351 of human cTnT gene. The upper lane showing sequences of homozygous wild type allele ‘C’ in a control individual. The middle and the lower lanes were showing the sequences of heterozygous (C/T  = Y) alleles in two individuals (a DCM patient and his relative, respectively). <b>Fig. 1B. G>A [IVS11-1G]</b> [rs483352835]: Electropherogram (arrow) showing a variant at splice acceptor site of human <i>cTnT</i> gene at nucleotide position g.16283, the electropherogram of a upper lane showing sequence of heterozygous (A/G = R) variant in a DCM patient, the lower lane showing sequence of control individual having wild type allele ‘G’ (homozygous). <b>Fig. 1C. N164N</b> [rs483352833]: Electropherogram (arrow) showing a novel synonumous mutation (N164) at the nucleotide position g.15304 of human <i>cTnT</i> gene in 2 DCM patients. The upper lane shows the sequences of heterozygous (C/T  = Y) transition in a DCM patient. The middle lane was the sequences of a control individual showing the wild type allele ‘C’ (homozygous). The lower lane sequences showing heterozygous (C/T  = Y) transition was from a 2^nd DCM patient. <b>Fig. 1D</b>. [rs3729842]: Electropherogram showing (arrow) a single nucleotide polymorphism at the nucleotide position g.10636 (C/T = Y) in intron 5 of human <i>cTnT</i> gene. The upper and the middle lanes were sequences showing heterozygous (C/T = Y) transition in DCM patients, the lower lane showing homozygous wild type (C/C) allele in a control individual. <b>Fig. 1E</b>. [rs3729845]: Electropherogram showing (arrow) at the nucleotide position g.13011 of human <i>cTnT</i> gene. The upper lane showing sequences of the heterozygous (A/G  = R) transition, and the lower lane showing homozygous wild type (G/G) allele of a control. <b>Fig. 1F</b>. [rs1104859]: Electropherogram showing (arrow) at the nucleotide position <b>g.11643</b> (A/C = M) in Intron 11 of human <i>cTnT</i> gene. The upper lane sequences showing the heterozygous (A/C = M) transversion, the middle lane showing homozygous wild type (G/G), and the lower lane sequences showing mutant homozygous (C/C) allele. <b>Fig.1G</b><b>. SNP-rs3729843</b>: Electropherogram showing (arrow) a SNP at the nucleotide position g.10822 (G/A = R) in intron 5 of human <i>cTnT</i> gene. The upper lane sequences showing mutant homozygous (A/A) allele. The middle lane sequences showing heterozygous (G/A = R) transition allele, and the lower lane showing sequences of homozygous wild type (G/G) allele in a control individual. <b>Fig. 1H</b>. [rs45576939]: Electropherogram showing (arrow) a novel mutation G>A at nucleotide position g.10370 in intron 4 of human <i>cTnT</i> gene, the upper lane displaying homozygous mutant (A/A) allele, and the lower lane showing sequences of a wild type allele (G/G). <b>Fig. 1I</b>. [rs45576635]: Electropherogram showing (arrow) a SNP at the nucleotide position g.14492 (C/T = Y) in intron 15 of human <i>cTnT</i> gene, the upper and the middle lanes sequences displaying heterozygous (C/T = Y) transition, and the lower lane sequences showing homozygous wild type (C/C) allele. <b>Fig. 1J</b>. [rs3729547]: Electropherogram showing (arrow) a polymorphic variant at the nucleotide position g.13424 of human <i>cTnT</i> gene, the upper lane displaying sequences of the heterozygous (C/T  = Y) transition, the middle lane sequences showing homozygous wild type (C/C) allele, and the lower lane displaying sequences of the homozygous mutant (T/T) allele. <b>Fig. 1K</b>. [rs483352834]. Electropherogram (arrow) showing a novel mutation at the nucleotide position g.15179 C>T in intron 11 of human <i>cTnT</i> gene, the upper lane displaying sequences of a DCM patient having heterozygous (C/T) transition, and the lower lane exhibiting sequences of a control individual having homozygous wild type allele (C/C). <b>Fig. 1L. K276K</b>. [rs483352836]: Electropherogram (arrow) exhibiting novel synonumous (K276) variant at the nucleotide position g.19429 of human <i>cTnT</i> gene in a DCM patient, the DCM patient displaying heterozygous (G/A  = R) transition. <b>Fig. 1M</b>. Sequence electropherogram showing (CTTCT) 5 bp Polymorphism. <b>Ma</b>. Presence of two copies of CTTCT (Insertion/Insertion – homozygous insertion) in both the chromosomes, <b>Mb</b>. Absence of one copy of CTTCT (Deletion/Deletion – homozygous deletion in both the chromosomes, <b>Mc</b>. Presence of 2 copies of CTTCT in one chromosome and presence of one copy of CTTCT in another chromosome (Insertion/deletion – heterozygous allele). g.6626-30 (5 bp).</p

Haplotypes on 9p21 modify the risk for coronary artery disease among Indians

The chromosomal region 9p21 has been reported to be associated with myocardial infarction, Coronary Artery Disease (CAD), diabetes and many other related multifactorial diseases in humans. Although the genome-wide association studies have identified a limited number of Single Nucleotide Polymorphisms (SNPs) at 9p21 for CAD risk, the role of flanking SNPs has not been studied so far. Therefore, in the present work, we studied the role of flanking SNPs with respect to that of the previously identified SNPs rs10757278 and rs2383207 at 9p21 among the Indian subjects found to have CAD (n = 414) along with age- and sex-matched control subjects (n = 408). Our study replicated the association of genome-wide association studies that had identified SNPs rs2383207 (p = 4.7 × 10⁻⁵) and rs10757278 (p = 5.5 × 10⁻⁵) among Indians with CAD. Further, we evaluated nine additional SNPs, of which two SNPs flanking rs2383207 (rs1537375 [p = 2.4 × 10⁻⁵] and rs1537374 [p = 5.6 × 10⁻⁵]) were also strongly associated with CAD. The haplotypes constructed using four risk SNPs revealed that the haplotypes with combinations of rs10757278 showed CAD risks, whereas the minor alleles of rs2383207, rs1537375, and rs1537374 in combinations reduce the CAD risks substantially. Our study demonstrates that the variation in the chromosomal region 9p21 is involved in modifying progression toward CAD among Indians and the risk may be variable, contributed by the SNPs that are flanking previously identified SNPs