A.P. Singh

Modulatory effect of areca nut on the action of mace (myristica fragrans, houtt) on the hepatic detoxification system in mice

The present paper reports the modifying potential of areca nut (Areca catechu), an ingredient of the habitual masticatory betel quid, on the induction of the hepatic detoxification system in mice by mace (the aril of nutmeg, Myristica fragrans) a known chemopreventor of chemically induced carcinogenesis. The modulatory effect of areca nut was assessed by determining the levels of enzymes of the hepatic detoxification system, such as glutathione S-transferase (GST), cytochrome b5 and cytochrome P-450, and the content of acid-soluble sulphhydryl (-SH). Mice were fed either control diet or diet containing 0.25, 0.5 or 1% areca nut for 45 days. During the last 10 days the diet was supplemented with 0.5 or 1% mace. At 0.5 and 1% in the diet, areca nut decreased mace-induced increases in hepatic GST and -SH levels and elevated further increases in the levels of cytochrome b5 and cytochrome P-450.

Modulatory potential of clocimum oil on mouse skin papillomagenesis and the xenobiotic detoxication system.

The present study was designed to elucidate the mechanistic inhibitory efficacy of clocimum (an eugenol rich variety of Ocimum gratissimum; Labiatae) oil on murine skin papillomagensis. Topical application of clocimum oil (50 microl/animal/day) during peri-initiation stage (1 week before and 2 weeks after initiation) of 7,12-dimethylbenz[a]anthracene (DMBA)-induced papillomagenesis and/or during the tumour promotion stage reduced (P < 0.05) the (i) tumour burden to 5.00, 4.41 and 4.50 (positive control value 5.27); (ii) cumulative number of papillomas to 85, 75 and 72 (positive control value : 95); and (iii) percent incidence of mice bearing papillomas to 94, 89 and 88, respectively (positive control value 94). Significant (P < 0.01) elevation in the hepatic levels of glutathione S-transferase (GST), sulfhydryl (-SH) and cytochrome b5 (Cyt. b5) was observed by the respective topical treatment of clocimum oil. Even in the skin tissue of the mouse, the topical treatment of clocimum oil enhanced (P < 0.05) the -SH content. The results suggest the modest chemopreventive potential of clocimum oil against the murine skin papillomagenesis, and such effects may be partly due to the modulated xenobiotic detoxication system enzymes.

Direct and translactational effect of arecoline alkaloid on the clocimum oil-modulated hepatic drug metabolizing enzymes in mice.

The present study assesses the potential of arecoline alkaloid, by direct exposure in lactating dams and translactational exposure in neonates, to modulate the efficacy of clocimum oil as a blocking agent in chemopreventive pathway. Clocimum oil (25 or 50 microl/dam/day) induced a significant increase in the hepatic levels of phase II glutathione S-transferase (GST) and acid-soluble sulfhydryl in lactating dams and suckling neonates while the elevated levels of hepatic phase I cytochrome b5 (Cyt. b5) and cytochrome P-450 (P450) were observed only in the dams. Arecoline (0.6 mg/dam/day) alone did not modulate the hepatic GST and sulfhydryl levels in either dams or pups, although significant induction was observed in the hepatic levels of Cyt. b5, P450 and malondialdehyde (MDA) in lactating dams and suckling neonates. Clocimum oil-modulated hepatic levels of phase II components were depressed whereas phase I enzymes and lipid peroxides levels were further elevated by clocimum oil-plus-arecoline treatment. The direct or translactationally augmented levels of bioactivated species of the administered compounds, via enhanced phase I oxidative catalysis and less efficient GST/GSH conjugational detoxication, may suggest the antagonistic influence of arecoline on chemopreventive efficacy of clocimum oil.

Inhibition of Infectious Bursal Disease Virus by Vector Delivered SiRNA in Cell Culture

Infectious Bursal Disease (IBD) is major threat to poultry industry. It causes severe immunosuppression and mortality in chicken generally at 3 to 6 weeks of age. RNA intereference (RNAi) emerges as a potent gene regulatory tool in last few years. The present study was conducted to evaluate the efficiency of RNAi to inhibit the IBD virus (IDBV) replication in-vitro. VP2 gene of virus encodes protein involved in capsid formation, cell entry and induction of protective immune responses against it. Thus, VP2 gene of IBDV is the candidate target for the molecular techniques applied for IBDV detection and inhibition assay. In this study, IBDV was isolated from field cases and confirmed by RT-PCR. The virus was then adapted on chicken embryo fibroblast cells (CEF) in which it showed severe cytopathic effects (CPE). The short hairpin RNA (shRNAs) constructs homologous to the VP2 gene were designed and one, having maximum score and fulfilling maximum Reynolds criteria, was selected for evaluation of effective inhibition. Selected shRNA construct (i.e., VP2-shRNA) was observed to be the most effective for inhibiting VP2 gene expression. Real time PCR analysis was performed to measure the relative expression of VP2 gene in different experimental groups. The VP2 gene was less expressed in virus infected cells co-transfected with VP2-shRNA as compared to mock transfected cells and IBDV+ cells (control) at dose 1.6 µg. The result showed ∼95% efficient down regulation of VP2 gene mRNA in VP2-shRNA treated cells. These findings suggested that designed shRNA construct achieved high level of inhibition of VP2 gene expression in-vitro.

Detection of very virulent infectious bursal disease virus from a field outbreak in Central India

In order to detect infectious bursal disease virus (IBDV), bursal tissue was collected from 10 IBD-suspected birds from a 30-day-old, IBDV-vaccinated commercial broiler chicken flock of 2000 birds exhibiting clinical signs suggestive of infectious bursal disease (IBD). The presence of IBDV was confirmed by partial amplification of the VP2 gene by reverse transcription and polymerase chain reaction. Isolates were identified as very virulent strains of IBDV (vvIBDV) by nucleotide sequence analysis. The comparison of the VP2 nucleotide sequences among the isolates revealed the presence of single-nucleotide polymorphisms in the VP2 gene of IBDV in the same flock. The comparative analysis indicated that these viruses were genetically close to the vvIBDVs previously detected in India. Our analysis provided information about the existence of vvIBDV in Central India.

Synthesis of new hybrid sorbent 2-mercaptobenzaldehyde SBA-15 and its application in solid phase extraction of Cd(II) from water and food samples

In this paper, a new mesoporous hybrid material was fabricated by anchoring 2-mercaptobenzaldehyde (2MB) onto the surface of SBA-15. The adsorbent (2MB-SBA-15) was characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), nitrogen adsorption–desorption isotherm measurements and thermogravimetric analysis (TGA) and applied as a sorbent in the preconcentration of Cd(II) in water samples and food samples. The effect of various analytical parameters such as pH, dose of sorbent, amount of eluent, sample flow rate and interfering ions was investigated. The preconcentration capacity of the prepared material under optimized conditions for Cd(II) was 0.94 ± 0.01 mmol g−1. The Cd collected onto the sorbent was eluted out with 10 mL of 1 M HNO3 and determined by AAS. The LOD, LOQ and preconcentration factor of the present method were found to be 6.93 × 10−6, 1.21 × 10−5 mM and 100, respectively. The relative standard deviation for four preconcentration experiments was found to be ≤5%.