A novel association of a polymorphism in the first intron of adiponectin gene with type 2 diabetes, obesity and hypoadiponectinemia in Asian Indians

Adiponectin is an adipose tissue specific protein that is decreased in subjects with obesity and type 2 diabetes. The objective of the present study was to examine whether variants in the regulatory regions of the adiponectin gene contribute to type 2 diabetes in Asian Indians. The study comprised of 2,000 normal glucose tolerant (NGT) and 2,000 type 2 diabetic, unrelated subjects randomly selected from the Chennai Urban Rural Epidemiology Study (CURES), in southern India. Fasting serum adiponectin levels were measured by radioimmunoassay. We identified two proximal promoter SNPs (−11377C→G and −11282T→C), one intronic SNP (+10211T→G) and one exonic SNP (+45T→G) by SSCP and direct sequencing in a pilot study (n = 500). The +10211T→G SNP alone was genotyped using PCR-RFLP in 4,000 study subjects. Logistic regression analysis revealed that subjects with TG genotype of +10211T→G had significantly higher risk for diabetes compared to TT genotype [Odds ratio 1.28; 95% Confidence Interval (CI) 1.07–1.54; P = 0.008]. However, no association with diabetes was observed with GG genotype (P = 0.22). Stratification of the study subjects based on BMI showed that the odds ratio for obesity for the TG genotype was 1.53 (95%CI 1.3–1.8; P < 10⁻⁷) and that for GG genotype, 2.10 (95% CI 1.3–3.3; P = 0.002). Among NGT subjects, the mean serum adiponectin levels were significantly lower among the GG (P = 0.007) and TG (P = 0.001) genotypes compared to TT genotype. Among Asian Indians there is an association of +10211T→G polymorphism in the first intron of the adiponectin gene with type 2 diabetes, obesity and hypoadiponectinemia.     

Expression analysis on mycoparasitism related genes during antagonism of <Emphasis Type="Italic">Trichoderma</Emphasis> with <Emphasis Type="Italic">Colletotrichum falcatum</Emphasis> causing red rot in sugarcane

Present study was aimed to select a suitable Trichoderma isolate as candidate antagonist based on its efficacy in producing cell wall degrading enzymes (CWDEs), its mycoparasitism activity and expression of related genes against the red rot pathogen caused by Colletotrichum falcatum in sugarcane. For which, six different isolates of Trichoderma selected from our earlier studies (T. harzianum, T. asperullum) were evaluated based on their capability in releasing cell wall degrading enzymes individually and during antagonism with C. falcatum in dual plate. Amongst T. harzianum (T20) exhibited the greatest mycoparasitic potential against the C. falcatum, by producing higher concentration of  CWDEs viz., chitinase and β-1, 3-glucanase, slightly lower amounts of cellulase and protease with significant reduction in polygalacturonase produced by pathogen. Further microscopic observation on interaction of C. falcatum with the selected isolate of T. harzianum (T20) exhibited the mycoparasitic activity of antagonist over pathogen in dual culture and inhibition of C. falcatum pathogenesis in detached sugarcane leaves. In addition, expression pattern of eight genes coding various enzymes involved in mycoparasitism by T. harzianum over C. falcatum were analyzed using qRT-PCR in vitro and on sugarcane leaves. In in vitro interactions, five genes of  cell wall degrading enzymes viz., chitinase (chit33), endochitinase (endo42), β-1, 3-glucanase (glu), exochitinase 1 (exc1), exochitinase 2 (exc2), were upregulated during and after contact as compared to before contact, while three genes related with proteases such as alkaline proteinase (prb1), trypsin-like protease (Pra1), subtilin-like serine protease (ssp), genes were upregulated during the contact with C. falcatum and slightly down regulated after contact. In detached leaves, seven genes were potentially upregulated except subtilin-like serine protease, which was down regulated during interaction of C. falcatum and T. harzianum as compared to T. harzianum inoculation alone. All these biochemical and molecular results confirm the efficacy of T. harzianum (T20) against C. falcatum and justify the right selection of candidate antagonist for our further studies on identification of antifungal genes/proteins against C. falcatum in sugarcane.     

Carbon-13 nmr spectra of vitamin B6 compounds: Aqueous solution equilibria and determination of microscopic acid dissociation constants

The aqueous solution equilibria and solute structure of vitamin B₆ compounds and several model compounds have been investigated using ¹³C-nmr spectroscopy. The unsubstituted α-carbon of these compounds is a very good probe for data which permits assignment of the ionization steps to indicidual groups. While the ionizations of the pyridinium and phenolic groups take place simultaneously in 3-hydroxypyridine, they take place in well-separated steps in pyridoxamine (PM), pyridoxamine phosphate (PMP), and pyridoxal phosphate. It has been established that the ionization with a pK_a value of 3.7 is predominantly phenolic in origin in PM and PMP. A zwitterionic structure consistent with the earlier spectroscopic investigations is proposed for the vitamin B₆ compounds in neutral aqueous solution.