Arsenic(III) sorption was investigated with nanostructured cerium incorporated manganese oxide (NCMO). The pH between 6.0 and 8.0 was optimized for the arsenic(III) sorption. Kinetics and equilibrium data (pH=7.0±0.2, T=303±1.6K, and I=0.01M) of arsenic(III) sorption by NCMO described, respectively, the pseudo-second order and the Freundlich isotherm equations well. The sorption process was somewhat complicated in nature and divided into two different segments, initially very fast sorption followed by slow intraparticle diffusion process. Sorption reaction of arsenic(III) on NCMO was endothermic (ΔH°=+13.46kJmol(-1)) and spontaneous (ΔG°=-24.75 to -30.15kJmol(-1) at T=283-323K), which took place with increasing entropy (ΔS°=+0.14kJmol(-1)K(-1)) at solid-liquid interface. Energy of arsenic(III) sorption estimated by analyzing the equilibrium data using the D-R isotherm model was 15.4kJmol(-1), indicating the ion-exchange type mechanism. Raman, FT-IR, pH effect, desorption, etc. studies indicated that arsenic(III) was oxidized to arsenic(V) during the sorption process.

Lung cancer is the leading cause of cancer dearth. Cigarette smoking is the strongest risk factor for developing lung cancer, which is conceivably initiated by proliferation. Here, we show that low concentration of aqueous extract of cigarette smoke (AECS) causes excessive proliferation of human lung epithelial cells (A549) without any apoptotic cell death. The causative factor responsible for AECS-induced proliferation has been identified as p-benzoquinone (p-BQ). Coimmunoprecipitation and immunoblot experiments indicate that p-BQ binds with epidermal growth factor receptor (EGFR). However, in contrast to EGF, it causes aberrant phosphorylation of EGFR that lacks c-Cbl-mediated ubiquitination and degradation resulting in persistent activation of EGFR. This is followed by activation of Hras + Kras and the downstream survival and proliferative signaling molecules Akt and ERK1/2, as well as the nuclear transcription factors c-Myc and c-Fos. Vitamin C and/or antibody to p-BQ prevents AECS/p-BQ-induced proliferation of lung cells apparently by inactivating p-BQ and thereby preventing activation of EGFR and the downstream signaling molecules. The results suggest that vitamin C and/or antibody to p-BQ may provide a novel intervention for preventing initiation of lung cancer in smokers.

A new dinuclear nickel(II) complex,[Ni(2)(LH(2))(H(2)O)(2)(OH)(NO(3))](NO(3))(3)(1), of an "end-off" compartmental ligand 2,6-bis(N-ethylpiperazine-iminomethyl)-4-methyl-phenolato, has been synthesized and structurally characterized. The X-ray single crystal structure analysis shows that the piperazine moieties assume the expected chair conformation and are protonated. The complex 1 exhibits versatile catalytic activities of biological significance, viz. catecholase, phosphatase, and DNA cleavage activities, etc. The catecholase activity of the complex observed is very dependent on the nature of the solvent. In acetonitrile medium, the complex is inactive to exhibit catecholase activity. On the other hand, in methanol, it catalyzes not only the oxidation of 3,5-di-tert-butylcatechol (3,5-DTBC) but also tetrachlorocatechol (TCC), a catechol which is very difficult to oxidize, under aerobic conditions. UV-vis spectroscopic investigation shows that TCC oxidation proceeds through the formation of an intermediate. The intermediate has been characterized by an electron spray ionizaton-mass spectrometry study, which suggests a bidentate rather than a monodentate mode of TCC coordination in that intermediate, and this proposition have been verified by density functional theory calculation. The complex also exhibits phosphatase (with substrate p-nitrophenylphosphate) and DNA cleavage activities. The DNA cleavage activity exhibited by complex 1 most probably proceeds through a hydroxyl radical pathway. The bioactivity study suggests the possible applications of complex 1 as a site specific recognition of DNA and/or as an anticancer agent.

A study was conducted to determine the effect of metals on soil microbial biomass and activities in landfill soils as well as normal background soil. The microbial biomass and activities were consistently higher in the landfill soils than in the background soil. Significant positive correlations existed between the microbial parameters and soil organic carbon. The landfill soils contained higher concentrations of metals (iron, manganese, copper, cadmium, lead and zinc) than did the background soil. Microbial parameters were negatively correlated with the metals, with inhibition increasing with the bioavailability of the metals. It is suggested that the metals affected microbial biomass and activities by behaving synergistically or additively with each other. Although the landfill soils had higher microbial biomass and activities than the background soil, due to higher organic matter content, the ratios of microbial parameters/organic carbon indicated that inhibition of microbial growth and activities had occurred due to metal stress.

Encapsidation of nascent genome RNA into an RNase-resistant form by nucleocapsid protein, N is a necessary step in the rhabdoviral life cycle. However, the precise mechanism for viral RNA specific yet processive encapsidation remains elusive. Using Chandipura virus as a model system, we examined RNA binding specificity of N protein and dissected the biochemical steps involved in the rhabdoviral encapsidation process. Our analysis suggested that N protein in its monomeric form specifically binds to the first half of the leader RNA in a 1:1 complex, whereas, oligomerization imparts a broad RNA binding specificity. We also observed that viral P protein and dissociating detergent deoxycholate, both were able to maintain N in a monomeric form and thus promote specific RNA recognition. Finally, use of a minigenome length RNA in an in vitro encapsidation assay revealed the monomeric N and not its oligomeric counterpart, to be the true encapsidating unit. Based on our observations, we propose a model to explain encapsidation that involves two discrete biochemically separable steps, initiation and elongation.

Cigarette smoke (CS) causes oxidative damage and tea polyphenols have strong antioxidant properties. Therefore, we studied the effect of a black tea (BT) infusion on CS-induced oxidative damage of proteins both in vitro and in vivo. In the in vitro experiment, bovine serum albumin (BSA) or a guinea pig tissue microsomal suspension was incubated with an aqueous extract of CS (CS-solution) in the presence or absence of the BT infusion. Protein oxidation was measured by immunoblotting of the dinitrophenylhydrazone derivatives of the protein carbonyls followed by densitometric scanning. Protein degradation was assessed by SDS-PAGE. BT prevented (P < 0.05) CS-induced oxidation of BSA and oxidative degradation of guinea pig lung, liver and heart microsomal proteins. This was also observed when the BT infusion was replaced by its components, i.e, flavonols, theaflavins, thearubigins and catechins. BT prevented microsomal protein degradation by inhibiting oxidative modification of the proteins. The antioxidant effect of BT was similar to that of green tea. In the in vivo experiment, partially ascorbate-deficient guinea pigs were subjected to CS exposure from 5 cigarettes/(guinea pig. d) for 7 d and given water or the BT infusion (20 g/L) to drink. Guinea pigs exposed to CS and given water had extensive oxidation accompanied by 39, 40 and 30% losses (P < 0.05) of microsomal proteins of lung, liver and heart, respectively. However, when the CS-exposed guinea pigs consumed the BT infusion instead of water, the oxidation of microsomal proteins was reduced (P < 0.05) approximately 90, 97 and 70% in lung, liver and heart, respectively. Protein loss was reduced (P < 0.05) approximately 92, 98 and 90% in lung, liver and heart, respectively. The results, if extrapolated to humans, would indicate that regular intake of tea may protect smokers from CS-induced oxidative damage and consequent degenerative diseases.

The molecular events associated with the transcriptive and replicative cycle of negative-stranded RNA viruses are still an enigma. We took Chandipura virus, a member of the Rhabdoviridae family, as our model system to demonstrate that Phosphoprotein P, besides Nucleocapsid protein N, also acts as a leader RNA-binding protein in its unphosphorylated form, whereas CKII-mediated phosphorylation totally abrogates its RNA-binding ability. However, interaction between P protein and leader RNA can be distinguished from N-mediated encapsidation of viral sequences. Furthermore, P protein bound to leader chain can successively recruit N protein on RNA while itself being replaced. We also observed that the accumulation of phosphorylation null mutant of P protein in cells results in enhanced genome RNA replication with concurrent increase in the viral yield. All these results led us to propose a model explaining viral transcription-replication switch where Phosphoprotein P acts as a modulator of genome transcription and replication by its ability to bind to the nascent leader RNA in its unphosphorylated form, promoting read-through of the transcription termination signals and initiating nucleocapsid assembly on the nascent RNA chain.

Our recent in vitro results [4] indicate that cigarette smoke induces oxidation of human plasma proteins and extensive oxidative degradation of the guinea pig lung, heart, and liver microsomal proteins, which is almost completely prevented by ascorbic acid. In this paper, we substantiate the in vitro results with in vivo observations. We demonstrate that exposure of subclinical or marginal vitamin C-deficient guinea pigs to cigarette smoke causes oxidation of plasma proteins as well as extensive oxidative degradation of the lung microsomal proteins. Cigarette smoke exposure also results in some discernible damage of the heart microsomal proteins. The oxidative damage has been manifested by SDS-PAGE, accumulation of carbonyl and bityrosine, as well as loss of tryptophan and protein thiols. Cigarette smoke exposure also induces peroxidation of microsomal lipids as evidenced by the formation of conjugated dienes, malondialdehyde, and fluorescent pigment. Cigarette smoke-induced oxidative damage of proteins and peroxidation of lipids are accompanied by marked drop in the tissue ascorbate levels. Protein damage and lipid peroxidation are also observed in cigarette smoke-exposed pair-fed guinea pigs receiving 5 mg vitamin C/animal/day. However, complete protection against protein damage and lipid peroxidation occurs when the guinea pigs are fed 15 mg vitamin C/animal/day. Also, the cigarette smoke-induced oxidative damage of proteins and lipid is reversed after discontinuation of cigarette smoke exposure accompanied by ascorbate therapy. The results, if extrapolated to humans, indicate that comparatively large doses of vitamin C may protect the smokers from cigarette smoke-induced oxidative damage and associated degenerative diseases.

The phosphoprotein P of Chandipura (CHP) virus, an Indian isolate of rhabdovirus, was found to support transcription upon phosphorylation by casein kinase II (CKII). A phosphorylation-induced change in the protein conformation was found to occur at the N-terminal region of the protein. Biochemical studies for further characterization of this phosphorylation-based conformational alteration demonstrated that phosphorylation leads to the transition from an 'open' to 'closed' structure of the protein. The phosphate group introduced by CKII was found to be resistant to phosphatases. This phosphorylation-based structural alteration changes the accessible hydrophobic surface area of the protein and also the available digestion sites of different proteases. The phosphorylated form of P protein was found to be a dimer by His-tag dilution assay. Using the same approach it was found that the N-terminal 46 amino acids are responsible for P-P dimerization, only after phosphorylation.

A PCR based detection that amplifies the 552-bp intergenic spacer (IGS) region of multicopy rRNA gene of Giardia lamblia and 320-bp internal sequences to first PCR product has been used in diagnosis of giardiasis in stool sample. The primers were found highly specific to Giardia spp. only, because no amplification was observed with DNAs from other enteric pathogens like Escherichia coli, Shigella dysenteriae and Entamoeba histolytica. The test could detect even less than 2 pg of genomic DNA from Giardia trophozoites. In direct diagnosis of Giardia lamblia in stool samples, it was observed that PCR amplification of IGS followed by nested PCR could enhance the sensitivity and specificity of the tests manifold and the system was able to detect as low as 10 parasites in 100 microl of stool. The comparative evaluation of the present system with conventional microscopy, CIEP and ELISA in the diagnosis of giardiasis from diarrhoeic stool samples and control subjects demonstrated a 100% correlation among nested PCR, microscopic examination and ELISA in patients suggestive of giardiasis (Group I) and control subjects (Group II). In Group I cases (patients suffering from other than giardiasis), CIEP, ELISA and nested PCR showed better results than microscopic examination. However, among them, PCR was found most sensitive and specific because 20% positivity was noticed by PCR whereas CIEP and ELISA showed only 7.14% and 12.85%, respectively. Break-up results showed that all the samples which were positive by CIEP or ELISA, also found positive by PCR. The present observation clearly suggests the use of PCR that amplifies the intergenic spacer region of multicopy rRNA gene of Giardia lamblia followed by nested PCR for routine, quick and reliable detection of Giardia lamblia in stool samples.