Inhibition of suicidal erythrocyte death by pyrogallol

Molecular Biology Reports - Pyrogallol, a polyphenolic component of Acacia nilotica has previously been reported to induce apoptosis of diverse cell types. Pyrogallol is in part effective by...     

The alkali molten globule state of ferrocytochrome c: extraordinary stability, persistent structure, and constrained overall dynamics

This paper describes the structural and dynamic properties of a hitherto uncovered alkali molten globule (MG) state of horse "ferrocytochrome c" (ferrocyt c). Several experimental difficulties mainly because of heme autoxidation and extraordinary stability of ferrocyt c have been overcome by working with the carbonmonoxide-bound molecule under extremely basic condition (pH 13) in a strictly anaerobic atmosphere. Structural and molecular properties extracted from basic spectroscopic experiments suggest that cations drive the base-denatured CO-liganded protein to the MG state. The stability of this state is approximately 5.2 kcal mol(-)(1), and the guanidinium-induced unfolding transition is sharp (m(g) approximately 2.3 kcal mol(-)(1) M(-)(1)), suggesting contents of rigid tertiary structure. Strategic experiments involving the measurement of the CO association rate to the base-denatured protein and intrachain diffusion rates measured by laser photolysis of CO indicate a substantially restricted overall motion and stiffness of the polypeptide chain in the MG state. Possible placement of the state in the folding coordinate of ferrocyt c is discussed.     

Membrane bound pyrophosphatase and P-Type adenosine triphosphatase of <Emphasis Type="Italic">Leishmania donovani</Emphasis> as possible chemotherapeutic targets: Similarities and differences in inhibitor sensitivities

The activities of inorganic pyrophosphatase (PPase) and adenosine triphosphatase (ATPase) were studied in the plasma membrane of Leishmania donovani promastigotes and amastigotes. It was shown that the specific activity of PPase was greater than that of ATPase in the promastigote plasma membrane. We characterized H⁺-PPase present in the plasma membrane of L. donovani and investigated its possible role in the survival of promastigote and amastigote. PPase activity was stimulated by K⁺ and sodium orthovanadate and inhibited by pyrophosphate analogs (imidodiphosphate and alendronate), KF, N,N′-dicyclohexylcarbodiimide (DCCD), thiol reagents (p-chloromercuribenzenesulfonate (PCMBS), N-ethylmaleimide (NEM), and phenylarsine oxide (PAO)), the ABC superfamily transport modulator verapamil, and also by the F₁F_o-ATPase inhibitor quercetin. ATPase activity was stimulated by K⁺ and verapamil, inhibited by DCCD, PCMBS, NEM, sodium azide, sodium orthovanadate, and quercetin, and was unaffected by PAO. We conclude that there are significant differences within promastigote, amastigote, and mammalian host in cytosolic pH homeostasis to merit the inclusion of PPase transporter as a putative target for rational drug design.     

Tree diversity and population structure in undisturbed and human-impacted stands of tropical wet evergreen forest in Arunachal Pradesh, Eastern Himalayas, India

Tree species richness, tree density, basal area, population structure and distribution pattern were investigated in undisturbed, mildly disturbed, moderately disturbed and highly disturbed stands of tropical wet evergreen forests of Arunachal Pradesh. The forest stands were selected based on the disturbance index (the basal area of the cut trees measured at ground level expressed as a fraction of the total basal area of all trees including felled ones): (i) undisturbed stand (0% disturbance index), (ii) mildly disturbed (20% disturbance index), (iii) moderately disturbed (40% disturbance index), and (iv) highly disturbed stand (70% disturbance index). Tree species richness varied along the disturbance gradient in different stands. The mildly disturbed stand showed the highest species richness (54 of 51 genera). Species richness was lowest (16 of 16 genera) in the highly disturbed stand. In the undisturbed stand, 47 species of 42 genera were recorded while in the moderately disturbed stand 42 species of 36 genera were found. The Shannon–Wiener diversity index for tree species ranged from 0.7 to 2.02 in all the stands. The highest tree diversity was recorded in the undisturbed stand and the lowest in the highly disturbed stand. The stands differed with respect to the tree species composition at the family and generic level. Fagaceae, Dipterocarpaceae and Clusiaceae dominated over other families and contributed 53% in the undisturbed, 51% in the mildly disturbed, 42% in the moderately disturbed and 49% in the highly disturbed forest stands to the total density of the respective stand. Stand density was highest (5452 stems ha^–1) in the undisturbed stand, followed by the mildly disturbed stand (5014), intermediate (3656) in the moderately disturbed stand and lowest (338) in the highly disturbed stand. Dominance, calculated as the importance value index of different species, varied greatly across the stands. The highest stand density and species richness were represented in the medium girth class (51–110 cm) in all the stands. In the undisturbed stand, the highest density was found in the 111–140 cm girth class, while in the mildly disturbed stand the 51–80 cm girth range recorded the highest density. About 55, 68 and 52% species were found to be regenerating in the undisturbed, mildly disturbed and moderately disturbed stands, respectively. No regeneration was recorded in the highly disturbed stand. Variation in species richness, distribution pattern and regeneration potential is related to human interference and the need for forest conservation is emphasized.     

Participation of chlorobiumquinone in the transplasma membrane electron transport system of Leishmania donovani promastigote: effect of near-ultraviolet light on the redox reaction of plasma membrane

The respiratory quinone composition of the parasitic protozoa Leishmania donovani promastigote was investigated. 1'-oxomenaquinone-7, a chlorobiumquinone was found to be the major isoprenoid quinone. Substantial level of ubiquinone-9 was also present. Isolation and identification of the quinone from the purified plasma membrane yielded mainly 1'-oxomenaquinone-7 and ubiquinone-9; menaquinone was not detected. Membrane bound 1'-oxomenaquinone-7 could be destroyed by near-ultraviolet irradiation, with a concomitant loss or stimulation of plasma membrane electron transport activities. The abilities of different quinones to restore alpha-lipoic acid and ferricyanide reductase activity in near UV-irradiated cell preparations were compared. The order was; conjugate of chlorobiumquinone and sphingosine base approximately conjugate of 2-methyl-3-(1'-oxooctadecyl)-1,4-napthoquinone and octadecylamine > chlorobiumquinone approximately 2-methyl-3-(1'-oxooctadecyl)-1,4-napthoquinone > menaquinone-4 approximately ubiquinone-10. After irradiation with near-UV light, transmembrane alpha-lipoic acid reduction was inhibited, while transmembrane ferricyanide reduction was stimulated. The result obtained indicates that chlorobiumquinone mediates the plasma membrane electron transport between cytosolic reductant and oxygen as well as alpha-lipoic acid. UV-inactivation of chlorobiumquinone shuts down the plasma membrane oxygen uptake and diverts the electron flux towards ferricyanide reduction via ubiquinone-9. Chlorobiumquinone is the only example of a polyisoprenoid quinone containing a side chain carbonyl group from photosynthetic green-sulphur bacteria. Recent work has revealed numerous genes of trypanosomatid sharing common ancestry with plants and/or bacteria. These observations pose some fascinating questions about the evolutionary biology of this important group of parasitic protozoa.     

Short interfering RNA-mediated inhibition of herpes simplex virus type 1 gene expression and function during infection of human keratinocytes

RNA interference (RNAi) is an antiviral mechanism that is activated when double-stranded RNA is cleaved into fragments, called short interfering RNA (siRNA), that prime an inducible gene silencing enzyme complex. We applied RNAi against a herpes simplex virus type 1 (HSV-1) gene, glycoprotein E, which mediates cell-to-cell spread and immune evasion. In an in vitro model of infection, human keratinocytes were transfected with siRNA specific for glycoprotein E and then infected with wild-type HSV-1. RNAi-mediated gene silencing reproduced the small plaque phenotype of a gE-deletion mutant virus. The specificity of gene targeting was demonstrated by flow cytometry and Northern blot analyses. Exogenous siRNA can suppress HSV-1 glycoprotein E expression and function during active infection in vitro through RNAi. This work establishes RNAi as a genetic tool for the study of HSV and provides a foundation for development of RNAi as a novel antiviral therapy.     

Isolation of colour components from native dye-bearing plants in northeastern India

Recently dyes derived from natural sources have emerged as important alternatives to synthetic dyes. A study was initiated in the year 2000 at the RRL (CSIR), Jorhat to extract dyes from parts of five different plant species indigenous to northeastern India. The colour components responsible for dyeing were isolated and their chemical constituents were established based on chemical and spectroscopic investigations. The principal colour components from the species Morinda angustifolia Roxb., Rubia cordifolia Linn. and Tectona grandis Linn. were found to contain mainly anthraquinone moieties in their molecules. Those from the species Mimusops elengi Linn. and Terminalia arjuna (Roxb.) Wight & Arn. contained flavonoid moieties in their molecules. The absorption of dye (%) on fibres increased with increasing concentrations of dye in the dye-bath. Maximum absorption of dyes on fibres was obtained at 3% concentration of dyes obtained from R. cordfolia (35.350%), M. angustifolia (31.580%) and T. grandis (25.888%) and at 4% concentration of the dyes from M. elengi (31.917%) and T. arjuna (12.246%). The K/S values were found to increase with the increase in concentration of mordants. The colour co-ordinates of dyed samples were found to lie in the yellow-red quadrant of the colour space diagram. The dyes obtained from the native plants may be alternative sources to synthetic dyes for the dyeing of natural silk and cotton.