Effects of gamma radiation on the early developmental stages of Zebrafish (Danio rerio).

The zebrafish is gaining importance as a popular vertebrate model organism and is widely employed in ecotoxicological studies, especially for the biomonitoring of pollution in water bodies. There is limited data on the genetic mechanisms governing the adverse health effects in regards to an early developmental exposure to gamma radiation. In the present study zebrafish (Danio rerio) embryos were exposed to 1, 2.5, 5, 7.5 and 10Gy of gamma radiation at 3h post fertilization (hpf). Different developmental toxicity endpoints were investigated. Further, expression of genes associated with the development and DNA damage i.e. (sox2 sox19a and p53) were evaluated using Quantitative PCR (qPCR). The significant changes in the expression of sox2 sox19a and p53 genes were observed. This data was supported the developmental defects observed in the zebrafish embryo exposed to gamma radiation such as i.e. increased DNA damage, decreased hatching rate, increase in median hatching time, decreased body length, increased mortality rate, increased morphological deformities. Further, study shows that the potential ecotoxicological threat of gamma radiation on the early developmental stages of zebrafish. Further, it revealed that the above parameters can be used as predictive biomarkers of gamma radiation exposure.

Tributyltin chloride (TBTCl)-enhanced exopolysaccharide and siderophore production in an estuarine Alcaligenes faecalis strain.

Tributyltin chloride (TBTCl) has been used extensively as an antifouling agent in ship paints, which results in the contamination of aquatic sites. These contaminated sites serve as enrichment areas for TBTCl-resistant bacterial strains. One TBTCl-resistant bacterial strain was isolated from the sediments of Zuari estuary, Goa, India, which is a major hub of various ship-building activities. Based on biochemical characteristics and 16S rDNA sequence analysis, this bacterial strain was identified as Alcaligenes faecalis and designated as strain SD5. It could degrade ≥3 mM TBTCl by using it as a sole carbon source and transform it into the less toxic dibutyltin chloride, which was confirmed by nuclear magnetic resonance and mass spectroscopy. Interestingly, this bacterial strain also showed enhanced exopolysaccharide and siderophore production when cells were exposed to toxic levels of TBTCl, suggesting their involvement in conferring resistance to this antifouling biocide as well as degradative capability respectively.

Rhizospheric fungal community structure of a Bt brinjal and a near isogenic variety.

AIMS: The objective of this study was to investigate the influence of Cry1Ac gene expressing brinjal (VRBT-8) on the rhizospheric fungal community structure. METHODS AND RESULTS: qPCR indicated variations in the fungal ITS rRNA copy numbers of non-Bt (1·43-4·43) × 10(9) g(-1) dws and Bt (1·43-3·32) × 10(9) g(-1) dws plots. Phylogenetic analysis of ITS rRNA clones indicated fungal-related group majority of being Ascomycota compared to that of Basidiomycota and Zygomycota in non-Bt- and Bt-planted soils. Sordariomycetes was the dominant class detected in all the stages. CONCLUSIONS: Despite the variations in the population size and the distribution pattern observed across the non-Bt and Bt brinjal, plant-growth-dependent variability was more prominent compared with genetic modification. Therefore, this study concludes that genetic modification of brinjal crop has minor effect on the fungal community. SIGNIFICANCE AND IMPACT OF THE STUDY: Brinjal, the important solanaceous crop, is also prone to attack by many insect pests, especially by Leucinoides orbonalis, resulting in significant losses in the crop yield. However, the reports on the effect of transgenic crops and the associated microbial community are inconsistent. The present communication takes into account for the first time the possible interactions between Bt brinjal and the associated fungal community; the latter playing a significant role in maintaining soil fertility. As this study is limited to the structural diversity of fungal community, additional information regarding the functional diversity of the group seems imperative before recommending the commercialization of GM crops.

Lead-resistant Providencia alcalifaciens strain 2EA bioprecipitates Pb+2 as lead phosphate.

A lead-resistant bacteria isolated from soil contaminated with car battery waste were identified as Providencia alcalifaciens based on biochemical characteristics, FAME profile and 16S rRNA sequencing and designated as strain 2EA. It resists lead nitrate up to 0·0014 mol l(-1) by precipitating soluble lead as insoluble light brown solid. Scanning electron microscopy coupled with energy-dispersive X-ray spectrometric analysis (SEM-EDX) and X-ray diffraction spectroscopy (XRD) revealed extracellular light brown precipitate as lead orthophosphate mineral, that is, Pb(9) (PO(4))(6) catalysed by phosphatase enzyme. This lead-resistant bacterial strain also demonstrated tolerance to high levels of cadmium and mercury along with multiple antibiotic resistance. Providencia alcalifaciens strain 2EA could be used for bioremediation of lead-contaminated environmental sites, as it can efficiently precipitate lead as lead phosphate.

Bronco T (Shirisadi kasaya), a polyherbal formulation prevents LPS induced septicemia in rats.

OBJECTIVE: Here, Bronco T (BT), a polyherbal formulation developed in 1984 for treating asthma, has been repurposed against septicemia-induced ALI. MATERIALS AND METHODS: Lipopolysaccharides (3 mg/kg BW) were injected intraperitoneally before 24 hours of surgery to assess the cardiorespiratory parameters, blood PaO2/FiO2 and MPO, pulmonary water content and histological changes in the lungs. The pentoxifylline (PTX) (25 mg/kg BW) was used as the positive control and given one hour before LPS. BT was given 3 hours (orally at different doses of 3, 1.5 and 0.75 g/kg BW) before LPS. RESULTS: The LPS treated group showed significant bradypnea, hypotension and bradycardia, through elongated peaks (RR) and (MAP) respectively and finally death after 95 minutes of LPS injection. The PTX and BT (3 g/kg BW) pretreatment significantly prevented these changes (dose-dependent in the BT group). The survival in these groups was maintained up to 190 min after LPS. The Pentoxifylline showed a better response (75%) than Bronco T (72%). In both the treatments, a significant decrease in pulmonary water content and minimal neutrophil infiltration and intact alveoli-capillary membrane was seen in the transverse section (T.S) of the lungs. CONCLUSIONS: Significant improvement was noted in survival time with lesser tissue damage and improved pulmonary function was observed by pre-treating with Bronco T in LPS induced septicemia.

Contamination of ground water as a consequence of land disposal of dye waste mixed sewage effluents: a case study of Panipat district of Haryana, India.

Spatial samples of surface and ground water collected from land disposal site of dye waste mixed sewage effluents at Binjhole, in Haryana, India were analyzed to evaluate its effect on quality of pond, hand pumps and ground waters for human health and irrigation purposes. It was found that average COD and TDS of dye houses discharge (310 and 3,920 mg/L) and treated sewage (428 and 1,470 mg/L) on mixing acquired the values of 245 and 1,780 mg/L and only Pb (0.24 microg/L) was above the permissible limit for irrigation purpose. Disposal of this mixed water to village pond changes the COD and TDS to 428 and 1,470 mg/L, respectively. COD and TDS of hand pump water samples were 264 and 1,190 mg/L, where as in tube well water these values were 151 and 900 mg/L. Though the ground water contamination seemed to decrease with the increasing distance from the pond but COD, TDS and BOD values continued to be quite high in water samples drawn from the hand pumps up to a distance of 500 m from pond. However, the major cause of the concern in these waters was Pb (0.11-0.45 ppm). Crops grown with this water shows accumulation of heavy metals like Pb,Cd, Fe, Mn, Ni, Cu, and Zn but in few crops they (Zn, Pb and Cd) exceed the safe limits. Regular consumption of these crop products may lead heavy metal toxicity. It was concluded from this study that the deep seepage of effluents led to deterioration of ground water quality for drinking purposes and the well waters rendered unfit for irrigation purposes within a span of 2 years. This warrants appropriate disposal measures for sewage and dye industry effluents in order to prevent deterioration of ground water and health of human and animals.

Molecular ecological perspective of methanogenic archaeal community in rice agroecosystem.

Methane leads to global warming owing to its warming potential higher than carbon dioxide (CO2). Rice fields represent the major source of methane (CH4) emission as the recent estimates range from 34 to 112 Tg CH4 per year. Biogenic methane is produced by anaerobic methanogenic archaea. Advances in high-throughput sequencing technologies and isolation methodologies enabled investigators to decipher methanogens to be unexpectedly diverse in phylogeny and ecology. Exploring the link between biogeochemical methane cycling and methanogen community dynamics can, therefore, provide a more effective mechanistic understanding of CH4 emission from rice fields. In this review, we summarize the current knowledge on the diversity and activity of methanogens, factors controlling their ecology, possible interactions between rice plants and methanogens, and their potential involvement in the source relationship of greenhouse gas emissions from rice fields.

Biofiltration of xylene using wood charcoal as the biofilter media under transient and high loading conditions.

The main objective of this study was to evaluate the performance of wood charcoal as biofilter media under transient and high loading condition. Biofiltration of xylene was investigated for 150days in a laboratory scale unit packed with wood charcoal and inoculated with mixed microbial culture at the xylene loading rates ranged from 12 to 553gm-3h-1. The kinetic analysis of the xylene revealed absence of substrate inhibition and possibility of achieving higher elimination under optimum condition. The pH, temperature, pressure drop and CO2 production rate were regularly monitored during the experiments. Throughout experimental period, the removal efficiency (RE) was found to be in the range of 65-98.7% and the maximum elimination capacity (EC) was 405.7gm-3h-1. Molecular characterization results show Bacillus sp. as dominating microbial group in the biofilm.

Mutagenicity and tumorigenicity of dihydrodiols, diol epoxides, and other derivatives of benzo(f)quinoline and benzo(h)quinoline.

The mutagenic activities of benzo[f]quinoline, benzo[h]quinoline, and a number of their derivatives, including dihydrodiols, K-region oxides, diol epoxides, and tetrahydroepoxides, were assessed in strain TA 100 of Salmonella typhimurium. The dihydrodiol derivatives of benzo[f]quinoline and benzo[h]quinoline were also tested for tumorigenic activity in newborn mice. Benzo[f]quinoline was metabolically activated in the presence of rat liver S-9 preparation to products mutagenic to the bacterial system to a greater extent than was benzo[h]quinoline. However, trans-7,8-dihydro-7,8-dihydroxybenzo[f]quinoline was less mutagenic compared to trans-7,8-dihydroxy-7,8-dihydrobenzo[h]quinoline in the presence of rat liver homogenate. The data on the mutagenic activity of the dihydrodiol derivatives of benzoquinolines were consistent with the intrinsic mutagenicity of the corresponding epoxide derivatives, in that the bay-region diol epoxides and tetrahydroepoxide of benzo[h]quinoline exhibited considerably higher mutagenic activities compared to those of the corresponding derivatives of benzo[f]quinoline at equivalent doses. The K-region oxides of benzo[f]quinoline and benzo[h]quinoline were significantly less mutagenic than their corresponding bay-region diol epoxide and tetrahydroepoxide derivatives. The demonstration that benzo[f]quinoline is significantly more mutagenic than trans-7,8-dihydro-7,8-dihydroxybenzo[f]quinoline, a precursor to the weakly mutagenic bay-region diol epoxide, suggests that the bay-region diol epoxide formation is not the principal pathway for the metabolic activation of benzo[f]quinoline to a mutagen. On the other hand, the isomeric benzo[h]quinoline appears to exert its mutagenic effect via the formation of its bay-region diol epoxide. These results indicate that the position of a nitrogen heteroatom in phenanthrene (the analogous carbocyclic aromatic hydrocarbon) not only has a marked effect on the mutagenic activities of the diol epoxide derivatives, but also can alter the metabolic activation pathways of the parent hydrocarbon. Benzo[f]quinoline, benzo[h]quinoline, and their dihydrodiol derivatives were not tumorigenic in newborn mice.

Design and characterization of microstrip based E-field sensor for GSM and UMTS frequency bands.

An Electric (E-) field sensor based on coplanar waveguide-fed microstrip antenna to measure E-field strength for dual-band operation at 914 MHz and 2.1 GHz is proposed, designed, and characterized. The parametric optimization of the design has been performed to obtain resonance at global system for mobile communication and universal mobile telecommunication system frequency band. Low return loss (-17 dB and -19 dB), appropriate gain (0.50 dB and 1.55 dB), and isotropic behaviour (directivity ∼ 1 dB), respectively, at 914 MHz and 2.1 GHz, are obtained for probing application. Antenna factor (AF) is used as an important parameter to characterize the performance of the E-field sensor. The AF measurement is explained in detail and results are reported. Finally, using the designed E-field sensor, the E-field strength measurements are carried out in a transverse electromagnetic cell. The key sources of uncertainties in the measurement are identified, evaluated, and incorporated into the final results. The measurement results are compared with theoretical values, which are found in good agreement. For comparative validation, the results are evaluated with reference to an already calibrated commercially available isotropic probe.

Enantioselective Tissue Distribution of Ketorolac and its Enantiomers in Rats.

The difference in tissue distribution of Ketorolac and its enantiomers were investigated in wistar rats. Separate high performance liquid chromatographic method was developed and validated for determination of Ketorolac and its enantiomers. Oyster BDS (150 × 4.6 mm id., 5 µm particle size) column was used for determination of concentration of Ketorolac. Ketorolac enantiomers were determined using Chiral-AGP column (100 × 4.0 mm I.D., particle size 5 µ, Chrom tech Ltd, Sweden). Detection was done at wavelength of 322 nm using an ultraviolet detector in the analytical system. Ketorolac enantiomers exhibit difference in their disposition in Wistar rats. In kidney, there was a significant difference in pharmacokinetic parameters. The Cmax was nearly 4 times and AUC 0-∞ was found to be more than double for S (-) Ketorolac than that of R (+) Ketorolac. MRT, Ke and t1/2 differ significantly in kidney. In liver, Cmax was found to be approximately 69% higher for S (-) Ketorolac compared to R (+) Ketorolac. AUC 0-∞ did not differ significantly for the enantiomers in liver. In liver, S (-) Ketorolac eliminated very fast in comparison to R (+) Ketorolac having t1/2 (one third) in comparison to R (+) Ketorolac. In lungs, there was no difference observed for Cmax and other parameters but AUC 0-∞ was found to be marginally higher for S (-) ketorolac.

Synthesis of substituted 1-hydroxy-2-naphthanilides as potential cestodicidal agents.

A series of substituted 1-hydroxy-2-naphthanilides 4--14 has been synthesized by treating 1-hydroxy-2-naphthoic acids 2 with substituted anilines 3. The nitronaphthanilides, on reduction and subsequent treatment with thiophosgene, gave the corresponding substituted 2-naphthanilide isothiocyanates 30--33. Substitution of the chlorine of 8 by various cyclic amines gave 3'-nitro-4'-substituted 1-hydroxy-2-naphthanilides 15--21. Various 3-aryl-4-oxo-2,3-dihydro-1,3-naphthoxazine-2-thiones 34-43 and 3 aryl-2,4-dioxo-2,3-dihydro-1,3-naphthoxazines 44--51 have been prepared by reacting the corresonding naphthanilides with thiophosgene and ethyl chloroformate, respectively. All the compounds were tested for their cestodicidal activity against Hymenolepis nana infection in rats; 30 was found to be the most active compound of the series, showing 100% clearance of infection at a single oral dose of 7.5 mg/kg.

Hepatic DNA adduct formation in rats treated with benzo[f]quinoline.

The formation of hepatic DNA adducts in male Sprague-Dawley rats following i.p. administration of benzo[f]quinoline (BfQ) was examined using a 32P-post-labeling assay. BfQ exhibited a low binding (11-27 amol adducts/microgram DNA) to liver DNA. Two BfQ-nucleoside adducts (one major and one minor) were detected. The BfQ-DNA adducts formed in vivo were chromatographically distinct from the adducts formed by the reaction of calf thymus DNA in vitro with BfQ-5,6-oxide, syn-7 beta,8 alpha-dihydroxy-9 beta,10 beta-epoxy-7,8,9,10-tetrahydroBfQ, anti-9 alpha,10 beta-dihydroxy-7 alpha,8 alpha-epoxy-7,8,9,10-tetrahydroBfQ, or anti-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydroBfQ-N- oxide. These results suggest that the bay-region diol epoxide of BfQ, unlike the bay-region diol epoxide derivatives of polynuclear aromatic hydrocarbons, is not involved in the covalent binding of BfQ to DNA.

Immunogenicity of three C-terminal synthetic peptides of the beta subunit of human chorionic gonadotropin and properties of the antibodies raised against 45-amino acid C-terminal peptide.

Immunological studies were carried out in rhesus monkeys and rabbits on three C-terminal synthetic peptides of beta-hCG (115-145; 111-145 and 101-145) after conjugating these to tetanus toxoid (TT). The immunogenicity of the peptide conjugates was comparatively poorer with reference to Pr-beta-hCG-TT conjugates at similar doses and immunization schedule. Amongst the three peptides, the best response was obtained with the 45-amino acid c-terminal peptide (45-CTP; 101-145). The anti-45-CTP recognized native hCG and was devoid of cross-reaction with hLH. hCG-induced testosterone production by mouse Leydig cells was inhibited by anti-45-CTP antiserum, although its neutralization capacity decreased more rapidly upon dilution than anti-beta-hCG sera of comparable titres. Immune complexes formed by the anti-45-CTP with hCG had a lower sedimentation value than those formed by anti-beta-hCG antisera with hCG, suggesting the presence of a limited number of immuno-determinant regions in the 45-amino acid C-terminal synthetic peptide.

Survival of animals injected with Pr-beta-HCG-TT.

PIP: 115 male Swiss strain mice were injected with Pr-B-HCG-TT vaccine and observed for 1 month. In another group of mice, a 2nd injection was given after 1 month and observation continued 8 weeks. Each injection was equivalent to 32 human doses. 6 of the 84 given 1 injection died; 1 of 31 given 2 injections died. A 10% mortality had been observed in the colony of untreated mice for this length of time. Autopsies showed no g ross or microscopic abnormalities. Anti-HCG antibodies were demonstrated in the animals' sera. 6 rabbits were each given 1-3 doses of the vaccine equivalent to 6-10 human doses. No deaths occurred up to 20 months of observation. 19 monkeys were each given 1-4 injections, each equivalent to 1 human dose, together with Freund's complete adjuvant or alum plus SO. At 2-16 months postinjections, survival was 100%. All animals tolerated the vaccine satisfactorily. Results show lack of toxicity of the vaccine.^ieng

Antibody response to Pr-beta-HCG-TT vaccine in human subjects.

PIP: Antibody response to processed beta human chorionic gonadotropin tetanus toxoid vaccine (Pr-beta-HCG-TT) was investigated in 4 human subjects who were in reproductive age and of proven fertility and had undergone tubal ligations. The subjects responded by producing detectable anti-HCG antibodies after a lag period of 6-8 weeks and plateau levels at about 5 months after primary immunization with the vaccine. Anti-HCG antibody titers were still high in 3 subjects after nearly 11 months of immunization. The titers declined to near 0 after 16 months in 1 subject indicating the reversibility of antibody response with this vaccine. The antibodies reacted immunologically with the HCG molecule and were competent to neutralize the biological activity of HCG in the radioligand receptor assay.^ieng

Processing of the preparations of beta-subunit of human chorionic gonadotropin for minimization of cross-reactivity with human luteinizing hormone.

PIP: The immunological reactivity of various components of beta-human cho rionic gonadotropin (beta-HCG) preparations with antisera developed against HCG and human luteinizing hormone (HLH) was assessed. Anti-beta-HCG reactivity was primarily located in 2 or 3 adjacent fractions. Anti-HLH reactivity was partly found in a fraction co-migrating with 1 of the anti-beta-HCG fractions, and also in a fraction migrating more slowly than the anti-beta-HCG fractions. Preparations with no, or minimal, cross-reactivity with anti-HLH were obtained upon absorption of beta-HCG with anti-ovine LH.^ieng

Detection of aromatic nitro compounds by means of their pi-complexes with N,N-diethylaniline.

The pi-complexes of nitro-aromatics are found to be more intensely coloured in the solid state than in solution. Thus nitro compounds, on spraying with N,N-diethylaniline on paper or thin-layer plates, yield yellow, red, brown or violet colours when the solvent is evaporated. The detection limit is 1-5 mug. Several donor groups do not vitiate the test when present as substituents.

Post-irrigation impact of domestic sewage effluent on composition of soils, crops and ground water--a case study.

Long-term irrigation with sewage water adds large amounts of carbon, major and micro- nutrients to the soil. We compared the spatial distribution of N, P, K and other micronutrients and toxic elements in the top 0.6 m of an alluvial soil along with their associated effects on the composition of crops and ground waters after about three decades of irrigation with domestic sewage effluent as a function of distance from the disposal point. Use of sewage for irrigation in various proportions improved the organic matter to 1.24-1.78% and fertility status of soils especially down to a distance of 1 km along the disposal channel. Build up in total N was up to 2908 kg ha(-1), available P (58 kg ha(-1)), total P (2115 kg ha(-1)), available K (305 kg ha(-1)) and total K (4712 kg ha(-1)) in surface 0.15 m soil. Vertical distribution of these parameters also varied, with most accumulations occurring in surface 0.3 m. Traces of NO3-N (up to 2.8 mg l(-1)), Pb (up to 0.35 mg l(-1)) and Mn (up to 0.23 mg l(-1)) could also be observed in well waters near the disposal point thus indicating initiation of ground water contamination. However, the contents of heavy metals in crops sampled from the area were below the permissible critical levels. Though the study confirms that the domestic sewage can effectively increase water resource for irrigation but there is a need for continuous monitoring of the concentrations of potentially toxic elements in soil, plants and ground water.

Heavy metal toxicity in a N2-fixing cyanobacterium, Anabaena doliolum: regulation of toxicity by certain environmental factors.

The toxic effects of chromium and tin salts on growth, 14C uptake, heterocyst differentiation, and nitrate reductase, nitrogenase, and glutamine synthetase activities of Anabaena doliolum and their regulation by pH, salinity, extracellular metabolites (spent), and organic acids have been studied. The toxicity of the test metals was lowered at alkaline pH and increased at acidic pH. NaCl at 20 mM was found to decrease metal toxicity. Extracellular metabolites (spent) in a 1:1 ratio (v/v) with fresh culture medium and organic acids were found to reduce metal toxicity. Among the various organic acids studied, humic acid was the most effective in regulating metal toxicity, apparently due to its multiple binding sites for metal cations. This study demonstrated that environmental factors, such as pH, salinity, extracellular metabolites, and organic acids, can mediate the toxicity of heavy metals to A. doliolum in a laboratory microcosm.