Serum proteomic signature of human chagasic patients for the identification of novel potential protein biomarkers of disease

Chagas disease is initiated upon infection by Trypanosoma cruzi. Among the health consequences is a decline in heart function, and the pathophysiological mechanisms underlying this manifestation are not well understood. To explore the possible mechanisms, we employed IgY LC10 affinity chromatography in conjunction with ProteomeLab PF2D and two-dimensional gel electrophoresis to resolve the proteome signature of high and low abundance serum proteins in chagasic patients. MALDI-TOF MS/MS analysis yielded 80 and 14 differentially expressed proteins associated with cardiomyopathy of chagasic and other etiologies, respectively. The extent of oxidative stress-induced carbonyl modifications of the differentially expressed proteins (n = 26) was increased and coupled with a depression of antioxidant proteins. Functional annotation of the top networks developed by ingenuity pathway analysis of proteome database identified dysregulation of inflammation/acute phase response signaling and lipid metabolism relevant to production of prostaglandins and arachidonic acid in chagasic patients. Overlay of the major networks identified prothrombin and plasminogen at a nodal position with connectivity to proteome signature indicative of heart disease (i.e., thrombosis, angiogenesis, vasodilatation of blood vessels or the aorta, and increased permeability of blood vessel and endothelial tubes), and inflammatory responses (e.g., platelet aggregation, complement activation, and phagocyte activation and migration). The detection of cardiac proteins (myosin light chain 2 and myosin heavy chain 11) and increased levels of vinculin and plasminogen provided a comprehensive set of biomarkers of cardiac muscle injury and development of clinical Chagas disease in human patients. These results provide an impetus for biomarker validation in large cohorts of clinically characterized chagasic patients.     

Enzymatic characterization of Catalase from Bacillus anthracis and prediction of critical residues using information theoretic measure of Relative Entropy

In order to cope up with the reactive oxygen species (ROS) generated by host innate immune response, most of the intracellular organisms express Catalase for the enzymatic destruction/detoxification of hydrogen peroxide, to combat its deleterious effects. Catalase thus, scavenges ROS thereby playing a pivotal role in facilitating the survival of the pathogen within the host, and thus contributes to its pathogenesis. Bacillus anthracis harbors five isoforms of Catalase, but none of them has been studied so far. Thus, this study is the first attempt to delineate the biochemical and functional characteristics of one of the isoforms of Catalase (Cat1.4) of B. anthracis, followed by identification of residues critical for catalysis. The general strategy used, so far for mutational analysis in Catalases is structure based, i.e. the residues in the vicinity of heme were mutated to decipher the enzymatic mechanism. However, in the present study, protein sequence analysis was used for the prediction of catalytically important residues of Catalase. Essential measures were adopted to ensure the accuracy of predictions like after retrieval of well-annotated sequences from the database with EC 1.11.1.6, preprocessing was done to remove irrelevant sequences. The method used for multiple alignment of sequences, was guided by structural alignment and thereafter, an information theoretic measure, Relative Entropy was used for the critical residue prediction. By exploiting this strategy, we identified two previously known essential residues, H55 and Y338 in the active site which were demonstrated to be crucial for the activity. We also identified six novel crucial residues (Q332, Y117, H215, W257, N376 and H146) located distantly from the active site. Thus, the present study highlights the significance of this methodology to identify not only those crucial residues which lie in the active site of Catalase, but also the residues located distantly.     

Chromium Removal from Aqueous System and Industrial Wastewater by Agricultural Wastes

This study involved the development of formaldehyde-treated, deseeded sunflower head waste–based biosorbent (FSH) for the biosorption of Cr(VI) from aqueous solution and industrial wastewater. Batch-mode experiments were conducted to determine the kinetics, sorption isotherms, effect of pH, initial Cr(VI) concentration, biosorbent dose, and contact time. The results demonstrated that FSH can sequester Cr(VI) from the aqueous solution. The maximum sorption occurred at pH = 2.0, biosorbent dose = 4.0 g/L, concentration of 100 mg/L at 25°C at 180 rpm after 2 h contact time. The FSH had an adsorption capacity of 7.85 mg/g for Cr(VI) removal at pH 2.0. The rate of adsorption was rapid, and equilibrium was attained within 2 h. The equilibrium sorption data fitted the Langmuir isotherm model, which was further confirmed by the chi-square test.     

Fatty acid desaturation in the intestinal mucosa

Information as to the ability of the enterocyte to desaturate fatty acids is lacking. This is important in understanding whether the source of intestinal arachidonic (20:4(n-6) acid is biliary or from de novo synthesis. Delta 9- and delta 6-desaturase enzymes were assayed in homogenates of rat jejunum, ileum and liver. Rat small intestine possesses desaturase activity to convert palmitic (16:0) to palmitoleic (16:1) and linoleic (18:2(n-6) to linolenic (18:3(n-6) acid. Enzyme activities were highest in liver relative to activity in jejunal and ileal homogenates. It is concluded that delta 9- and delta 6-desaturase activities may have an important role in determining physico-chemical properties and thus transport properties of enterocyte membranes.     

Influence of Water Stress on Water Relations, Photosynthetic Parameters and Nitrogen Metabolism of Moth Bean Genotypes

Effects of water stress at pre-flowering stage were studied in three genotypes (RMO-40, Maru moth and CZM-32 E) of moth bean [Vigna aconitifolia (Jacq.) Marechal]. Increasing water stress progressively decreased plant water potential, leaf area, net photosynthetic rate, starch and soluble protein contents and nitrate reductase activity while contents of reducing sugars, total soluble sugar, free amino acids and free proline progressively increased. Significant genotypic differences were observed and genotype CZM-32-E displayed a better drought tolerance than other genotypes.     

The C-terminal domain (CTD) in linker histones antagonizes anti-apoptotic proteins to modulate apoptotic outcomes at the mitochondrion

The loss of mitochondrial integrity as a consequence of apoptogenic complexes formed on the outer membrane constitutes a key step in controlling progression of apoptotic cascades. Here, we show that multiple members of the linker histone (LH) family of proteins modify apoptotic cascades initiated by the Bcl-2 protein Bak, and impart resistance to its endogenous antagonist Bcl-xL. Our experiments reveal apoptogenic capabilities equivalent to those documented for H1.2 in H1.1 and H1.3 isoforms. Deletion mutants of H1.2 and site-directed mutagenesis of H1.1 and H1.2 implicated the C-terminal domain in apoptogenic activity. In this context, disruption of protein kinase-C activity using chemical inhibitors, dominant-negative approaches and RNA interference coupled with site-directed modifications in H1.1, identified the protein kinase-Cβ1 isoform as a repressor of H1.1/H1.3 apoptogenic activity. Finally, a H1.2 C-terminal tail recombinant attenuated Bcl-xl inhibition of Bak-induced apoptosis, suggesting that the C-terminal domain was necessary and sufficient for apoptogenic functions. Thus, integration with apoptotic intermediates (via C-terminal tail interactions) may constitute a more generalized function of LH isoforms in apoptotic cascades.     

Seedling age and cytokinin effects on glutamate dehydrogenase activity and nitrogen assimilation in maize leaves

Glutamate dehydrogenase (GDH) activity, protein and total nitrogen contents in the secondary leaves of maize(Zea mays L. cv. Ganga Safed-2) seedlings increased during early seedling growth and then declined after reaching a peak level at either 10 d (GDH) or 12 d (metabolites). While the effect of kinetin on enzyme activity was statistically insignificant, benzyladenine supplied with nutrient solution increased GDH activity in secondary leaves of both 10-d as well as 14-d seedlings. However, both growth regulators increased the contents of total soluble proteins, total nitrogen, chlorophyll(a+b) and carotenoids in both 10 and 14-d old leaves.     

Restrictive localization of centromeric heterochromatin (C-bands) in Presbytis e. entellus (Dufresne) as compared to Macaca mulatta (Zimmerman)

C-bands are observed in the centromeric regions of only three pairs of autosomes and the distal portion of the small acrocentric Y in a total complement of 44 chromosomes of a male Presbytis e. entellus. Simultaneously treated slides of a Rhesus monkey, however, have C-bands in all the 42 chromosomes. The lack of C-bands may be due to (1) absence of highly repetitive DNA in the centromeric region of certain chromosomes or (2) presence of minute quantity of such DNA which is imperceptible or (3) different types of centromeric heterochromatin with a varying degree of repetition of DNA sequences all of which do not react in similar manner to various techniques employed at present. It is hypothesized that the centromeric heterochromatin rich in satellite DNA helps in withstanding the force of excessive coiling of chromosomes at the centromere to facilitate the functioning of the genes for microtubular protein during cell division when other genes are rendered inactive due to compactness of chromosomes.