Lack of interaction of circulating T cells with phytohemagglutinin in bacillary positive untreated lepromatous leprosy patients--identification of subpopulation of lymphocytes by shifts in electrophoretic mobility.

Incubation of human peripheral blood lymphocytes from normal healthy subjects with phytohamagglutinin (PHA), causes the reduction of the surface charge of a subpopulation of T cells by 1363 +/- 242 e.s.u./cm2. The affected subpopulation was predominantly the high charge-bearing cells identifiable with early (10 min) rosette-forming cells with sheep erythrocytes. Purified lymphocytes obtained from untreated bacillary-positive, lepromatous leprosy patients contained high charge-bearing T lymphocyte subpopulation. However, incubation with PHA did not result in the shift of electrophoretic mobility of these cells, suggesting the absence of interacting sites for the mitogen on the surface of these cells. The absence of mitogen-interacting sites is not an inherent trait of leprosy patients; the surface charge of lymphocytes from Dapsone-treated bacillary-negative subjects was reduced upon incubation with PHA. A close correlation was found between the number of cells whose charge alters on incubation with PHA and the transformation index obtained with this mitogen.     

Biochemical analysis of jejunal brush border membrane of golden hamster: pathogenic modulations due to ancylostomiasis.

The common hookworm (Ancylostoma ceylanicum) infection of humans was studied in golden hamsters model system. Significant biochemical modulations were observed in hamster jejunal brush border membrane (BBM), the primary site of infection. Analysis of BBM at the peak of infection (3-weeks) revealed a marked decrease in the activities of sucrase, lactase and maltase, while activities of alkaline phosphatase, (Ca2+ + Mg2+)-ATPase and gamma-glutamyl transpeptidase were increased. Kinetic studies conducted with maltase, a superficially localised enzyme of jejunal BBM, revealed loss of enzyme active site during the infection. Among other constituents, the levels of cholesterol and triglycerides were significantly decreased with slight increase in phospholipid content in the infected animals. The hookworm infection also caused a decline in total hexose content indicating an altered membrane glycocalyx. Conversely, there was significant enhancement of hydroxyproline and sialic acid contents. SDS-PAGE analysis showed an enhancement in both low and high molecular weight proteins in jejunal BBM preparations of the infected group. Gel electrophoresis of glycoproteins further revealed the appearance of two additional peaks in the low molecular weight region and concomitant disappearance of a peak in the high molecular weight region. These results strongly support the view that the hookworm infection causes severe damage not to the site of attachment alone but also to the entire cell lining of the jejunum and therefore could influence overall digestion and absorption.     

A natural product inspired hybrid approach towards the synthesis of novel pentamidine based scaffolds as potential anti-parasitic agents

A natural product inspired molecular hybridization approach led us to a series of novel pentamidine based pyrimidine and chalcone scaffolds. All the hybrids were evaluated for their anti-leishmanial potential. Most of the screened compounds have showed significant in vitro anti-leishmanial activity with less cytotoxicity in comparison to the standard drugs (pentamidine, sodium stibogluconate, and miltefosine). Additionally, anti-malarial screening of these compounds was also done and four compounds have shown superior activity against chloroquine resistance strain (K1) of Plasmodium falciparum.     

Calpain from rat intestinal epithelial cells: Age-dependent dynamics during cell differentiation

Micromolar and millimolar Ca²⁺-requiring neutral protease (calpain I and calpain II) along with their endogenous inhibitor calpastatin were isolated and partially purified from the same preparation of rat intestinal epithelial cells. Calpain I and II were partially purified by 1300 and 900-fold with 57 and 53 per cent yield, respectively. The optimum assay conditions revealed pH 7.5, 20 min incubation at 25° C and 0.24% casein substrate for both calpains. The optimum calcium concentration obtained for calpain I and II were 25 M and 4 mM, respectively. Distribution of rat intestinal epithelial cells calpain I and II along with calpastatin during cell differentiation stages in weanling to senescence age were studied. Calpain I in weanling rats was in an increasing order from villus to crypt regions. Adult rats indicated well expressed consistent calpain I throughout the differentiation stages. Whereas, significant lowering towards crypt region cells were evident in old rats. Calpain II in weanling and adult rats was found to be consistent throughout the differentiation stages. Old animals revealed an increasing trend from villus to crypt region with insignificant activity present in upper villus cells. Concomitantly, different concentrations of calpastatin were observed throughout the differentiation stages in all the age groups. Moreover, the levels of calpains exceeded that of calpastatin in most of the epithelial cell populations during developmental stages. In addition to casein, intestinal epithelial cell membranes were found to be equally good substrates for calpains. Proteolytic susceptibility of weanling, adult and old rat membrane proteins varied significantly all along the ageing process in rats. Simultaneous age-dependent calpastatin response were also evident. Taken together the results obtained provided strong evidence that calpain plays significant role in rat intestinal cell differentiation and ageing process with calpastatin as its specific regulatory protein.Abbreviations DEAE-cellulose O-(Diethylaminoethyl)-cellulose - EDTA Ethylene Diamine Tetra Acetic Acid - Tris Tris (hydroxymethyl) amino methane - KH₂PO₄ potassium dihydrogen orthophosphate - Na₂HPO₄ disodium hydrogen phosphate - CaCl₂ Calcium Chloride - TCA Trichloroacetic Acid - PMSF Phenylmethylsulfonyl Fluoride     

WRKY transcription factors: evolution,regulation, and functional diversity in plants

Protoplasma - The recent advancements in sequencing technologies and informatic tools promoted a paradigm shift to decipher the hidden biological mysteries and transformed the biological issues...     

Aqua mediated synthesis of substituted 2-amino-4H-chromenes and in vitro study as antibacterial agents

A simple, clean, environmentally benign route to the synthesis of 2-amino-chromenes is described using K2CO3 as a green catalyst in water under microwave irradiation. This implies a convenient route avoiding the usage of hazardous organic solvents and organic bases. This technique requires only water in both the reaction step and workup, thus rendering the whole procedure into a truly ecofriendly green protocol. All the synthesized compounds were shown to possess antibacterial activity as tested in vitro against standard strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus.     

Cytochemical localization of phosphatase in differentiating secondary vascular cells

Summary Phosphatase activity was studied in the cambium and differentiating vascular cells of beech by using a modified Washstein and Meisel method. After fixation in glutaraldehyde or crotonaldehyde and incubation in a medium containing ATP and lead nitrate at pH 7.2, a deposit of electron-opaque granules was found in the nucleoli, nucleoplasm, nuclear envelope, endoplasmic reticulum, plastids, mitochondria, and at the plasmalemma. Although located at these different sites, the distribution varied both inter- and intra-cellularly. This is thought to be a true reflection of the variation in activity between closely adjacent cells in this part of the stem.Some reaction was obtained when ADP replaced ATP in the reaction mixture, but there was no reaction at all when both ATP and ADP were omitted. Fixation in hydroxyadipaldehyde, or incubation at pH 6.4 both produced very little reaction.     

Diverse Secreted Effectors Are Required for Salmonella Persistence in a Mouse Infection Model

Salmonella enterica serovar Typhimurium causes typhoid-like disease in mice and is a model of typhoid fever in humans. One of the hallmarks of typhoid is persistence, the ability of the bacteria to survive in the host weeks after infection. Virulence factors called effectors facilitate this process by direct transfer to the cytoplasm of infected cells thereby subverting cellular processes. Secretion of effectors to the cell cytoplasm takes place through multiple routes, including two separate type III secretion (T3SS) apparati as well as outer membrane vesicles. The two T3SS are encoded on separate pathogenicity islands, SPI-1 and -2, with SPI-1 more strongly associated with the intestinal phase of infection, and SPI-2 with the systemic phase. Both T3SS are required for persistence, but the effectors required have not been systematically evaluated. In this study, mutations in 48 described effectors were tested for persistence. We replaced each effector with a specific DNA barcode sequence by allelic exchange and co-infected with a wild-type reference to calculate the ratio of wild-type parent to mutant at different times after infection. The competitive index (CI) was determined by quantitative PCR in which primers that correspond to the barcode were used for amplification. Mutations in all but seven effectors reduced persistence demonstrating that most effectors were required. One exception was CigR, a recently discovered effector that is widely conserved throughout enteric bacteria. Deletion of cigR increased lethality, suggesting that it may be an anti-virulence factor. The fact that almost all Salmonella effectors are required for persistence argues against redundant functions. This is different from effector repertoires in other intracellular pathogens such as Legionella.