Determination of Multi Elements in Tobacco Plant of Northeast India by Neutron Activation Analysis and Atomic Absorption Spectrometry.

Even when cultivated in uncontaminated soils, tobacco plant has higher propensity to extract and accumulate trace elements. The concentrations (mass fractions) of essential elements (K, Ca, Mg, Na, Cl, Mn, Fe, Cu, and Zn) and 28 non-essential elements in tobacco plant (leaves, stem, and root) of Northeast India and their respective soils were quantitatively measured. Hg mass fraction in all samples analyzed were found to be < 10 mg/kg. The heavy element mass fractions of tobacco are weakly correlated to different soil parameters. The bioconcentration factor values indicated that Cd (7) is selectively absorbed and translocated in the tobacco leaves compared to Zn (1.7), Cu (1.5), Ni (0.12), and Pb (0.1). Under acidic soil conditions, tobacco plant efficiently absorbed and translocated Cl- ion with great ease, whereas it may be a very low accumulator of rare-earth elements. The mass fractions of Mn, Cu, Sb, Cs, Rb, and Pb are very similar to the "reference plant," whereas significantly higher mass fractions of Al, Sc, Ti, Zr, Hf, Ta, Th, and U are present in the roots of tobacco plant relative to the "reference plant." Principal component analysis has revealed that Northeast Indian tobacco can be clearly differentiated from other varieties of tobaccos used in different countries because of their element profiles.

Reduction of hexavalent chromium by Exiguobacterium mexicanum isolated from chromite mines soil.

Hexavalent chromium is a highly toxic element generated due to indiscriminate chromite mining in Sukinda, Odisha. In the present research investigation a relatively higher Cr(VI) resistant (900 mg L-1) bacterium CWB-54 was isolated from the chromite mine water. Based on the biochemical and molecular analysis the strain (CWB-54) was identified as Exiguobacterium mexicanum. When this bacterium was grown at 35 °C, 100 rpm, pH~8.0, and fructose as an electron donor, it could reduce the total hexavalent chromium (100 mg L-1) supplemented in the medium within 33 h of incubation period. Though experiment was carried out to study the effect of Mn, Ni, Cd, Hg and Zn on Cr(VI) reduction by the strain E. mexicanum it has been observed that in the presence of Cd and Hg, Cr(VI) reduction drastically decreased. Characterization of Cr(VI) reduced product by SEM-EDX and TEM analysis revealed intracellular and extracellular Cr(III) deposition in the bacterium, which is assumed to be Cr(OH)3 precipitate in nanometric size. But the extracellular chromate reductase enzyme production is found to be negligible as compared to the intracellular enzyme production. The increased concentration of Cr(VI) above (1000 mg L-1) also showed the genotoxic effect on the DNA. Several reports have been published on Exiguobacterium sp. on different scientific aspect but the current report on the reduction of toxic Cr(VI) by a new species E. mexicanum is a novel one which established the potentiality of this microorganism for a broad area of application.

Macro-, micro-, and trace element distributions in areca nut, husk, and soil of northeast India.

In areca nut and husk, 14 elements (As, Ca, Cd, Cl, Co, Cu, K, Mg, Mn, Na, Rb, Sb, and Zn) were determined, while 34 elements including rare earth elements were detected in the corresponding soil samples using instrumental neutron activation analysis and atomic absorption spectrometry methods, whereas the concentration levels of Hg in tested samples are negligible, perhaps, below the detection limits. No rare earth elements were detected in edible areca nut. The concentration levels of various essential elements and heavy elements such as As, Cd, and Cu present in areca nut are within the permissible levels, whereas Pb content is relatively higher than FAO/WHO's permissible levels. The order of bioaccumulation index for heavy metals in areca nut was Cd > Sb > Cu > Zn ≥ Mn ≥ Co > Pb ≥ As. Bioaccumulation index values are indicating that areca palm may not be able to accumulate other heavy elements in the edible areca nut, except for Cd. On the basis of pollution indices, Northeast Indian soil may be relatively unpolluted.

Saraca asoca seed extract treatment recovers the trace elements imbalances in experimental murine visceral leishmaniasis.

Saraca asoca is an important plant species of India having variety of medicinal activity such as antiviral, anti-diabetic, antimicrobial, anti-inflammatory, anti-cancer etc. Indian Kala-azar (KA) or visceral leishmaniasis (VL) is a protozoan parasitic disease caused by Leishmania sp and is endemic in Indian subcontinent. VL mainly targets the poorest people who have been suffering from deficiency in protein, nutrients and essential trace elements which ultimately leads to immunodeficiency. Essential trace element, Zinc (Zn) controls multiple aspects of innate and adaptive immunity while Iron (Fe) is required for various cellular activities. Bromine (Br) is important for assembly of collagen IV scaffolds in tissue development and helps in signalling and Copper (Cu) performs several functions related to immune system. Intra-cardiac blood was collected from the experimental BALB/c mice groups including (a) healthy control, (b) infected control, (c) Saraca asoca seed extract (Sa-SE) treated groups. The trace elements level in blood of mice was measured by Energy Dispersive X-Ray Fluorescence technique. Interestingly, the decreased level of Zn, Fe and Br as well as increased level of Cu in diseased state came back to almost normal range upon treatment with Sa-SE. The trace elements imbalances thus were almost restored to normalcy by treating the experimental BALB/c mice with ethanolic seed extract of Saraca asoca.

Spatial and seasonal distribution of multi-elements in suspended particulate matter (SPM) in tidally dominated Hooghly river estuary and their ecotoxicological relevance.

The present work represented first study of the spatio-seasonal distribution of the multi-elements in the suspended particulate matter (SPM) of the tropical Hooghly river estuary (HRE), eastern part of India. The high load of SPM (20-3460 mg/l) might have induced negative impact on the phytoplankton density. The relative abundance of the studied elements exhibited the following decreasing trend (concentration in µg/g and %): Si(26.44 ± 3.75%) > Al(7.94 ± 1.52%) > Fe(6.17 ± 1.9%) > K(3.05 ± 1.5%) > Ca(1.97 ± 1.11%) > Mg(1.57 ± 1.71%) > Na(1.45 ± 8.40%) > Mn(1273 ± 2003) > Zn(178.43 ± 130.95) > V(151.54 ± 27.13) > Cr(147.08 ± 32.21) > Cu(62.06 ± 14.03) > Ni(49.64 ± 12.09) > Pb(21.5 ± 10.45). The accumulation of Ni, Cr, Pb, and Cu is mainly controlled by the formation of Fe hydroxides along with particulate organic carbon (POC) and salinity. The average geo-accumulation index (Igeo) and enrichment factor (EF) endorsed the substantial input of Cr (Igeo = 0.037; EF = 1.61) and Zn (Igeo = 0.123; EF = 2.07) from diffused pollution sources. From ecotoxicological point of view, the quality guidelines (QGs) suggested that Cr and Ni might possess frequent adverse biological effects. However, the mean probable effect level (PEL) quotient values revealed 49% probability of toxicity to the aquatic biota for five toxic elements (Cr, Ni, Cu, Zn, and Pb). The geochemical approaches, pollution indices, and statistical evaluation together revealed low to moderate contamination in the estuary. This baseline data would be beneficial in adopting proper management strategies for sustainable utilization and restoration of the water resources. The authors strongly recommend continuous systematic monitoring and installation of treatment plants for management of this stressed estuary.

Elemental distribution in urban sediments of small waterbodies and its implications: a case study from Kolkata, India.

The impacts of elemental pollution in sediments of freshwater bodies are of particular concern in rapidly urbanizing cities of the developing world and have been extensively studied in rivers and lakes. The current study is an attempt to highlight the importance of smaller waterbodies, which happen to form a natural network in cities, for assessing the contamination status of sediments. The distribution of elements (Al, Ca, Fe, K, Mg, S, Si, Ti, Ba, Mn, Sr, V, As, Cr, Cu, Ni, Pb and Zn) in sediments of 15 ponds and 6 canals was studied to understand the overall pollution status and the associated ecological risk to aquatic organisms. Geochemical indices revealed Cr, Cu, Pb and Zn to be the principal elements of concern. The mean concentration of Cr, Cu, Pb and Zn was 308, 174, 76.9 and 446 mg kg-1, respectively. Ecological risk assessment revealed that Cr in 86% sites, Ni in 52% sites, Cu and Zn in 28% sites and Pb in 10% sites were associated with possible ecological toxicity. The findings suggest that multielemental concentration in sediments of ponds and canals could effectively distinguish between pristine and polluted sites and suitably identify the main elements of concern to support cost-efficient waste management solutions customized to both the sites and elements of concern.

Biochemical, molecular, and elemental profiling of Withania somnifera L. with response to zinc stress.

Zn stress seriously induces various toxic responses in Withania somnifera L., when accumulated above the threshold level which was confirmed by investigating the responses of protein, expression of antioxidant enzymes, and elemental profiling on accumulation of Zn. Zn was supplemented in the form of ZnSO4 (0, 25, 50, 100, and 200 µM) through MS liquid medium and allowed to grow the in vitro germinated plants for 7 and 14 days. The study revealed that when the application of Zn increased, a significant reduction of growth characteristics was noticed with alterations of proteins (both disappearance and de novo synthesis). The activity of CAT, SOD, and GPX were increased up to certain concentrations and then declined, which confirmed through in-gel activity under different treatments. RT-PCR was conducted by taking three sets of genes from CAT (RsCat, Catalase1, Cat1) and SOD (SodCp, TaSOD1.2, MnSOD) and found that gene RsCat from CAT and MnSOD from SOD have shown maximum expression of desired genes under Zn stress, which indicate plant's stress tolerance mechanisms. The proton-induced X-ray emission study confirmed an increasing order of uptake of Zn in plants by suppressing and expressing other elemental constituents which cause metal homeostasis. This study provides insights into molecular mechanisms associated with Zn causing toxicity to plants; however, cellular and subcellular studies are essential to explore molecule-molecule interaction during Zn stress in plants.

Synthesis of a novel glucose capped gold nanoparticle as a better theranostic candidate.

Gold nanoparticles are predominantly used in diagnostics, therapeutics and biomedical applications. The present study has been designed to synthesize differently capped gold nanoparticles (AuNps) by a simple, one-step, room temperature procedure and to evaluate the potential of these AuNps for biomedical applications. The AuNps are capped with glucose, 2-deoxy-D-glucose (2DG) and citrate using different reducing agents. This is the first report of synthesis of 2DG-AuNp by the simple room temperature method. The synthesized gold nanoparticles are characterized with UV-Visible Spectroscopy, Fourier transform infrared spectroscopy (FTIR), Transmission electron microscopy (TEM) and selected area electron diffraction (SAED), Dynamic light scattering (DLS), and Energy-dispersive X-ray spectroscopy (SEM-EDS). Surface-enhanced Raman scattering (SERS) study of the synthesized AuNps shows increase in Raman signals up to 50 times using 2DG. 3-(4, 5-dimethylthiozol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay has been performed using all the three differently capped AuNps in different cell lines to assess cytotoxcity if any, of the nanoparticles. The study shows that 2DG-AuNps is a better candidate for theranostic application.

Particle induced X-ray emission study of blood samples of Indian Kala-azar patients.

Visceral leishmaniasis (VL) or Kala-azar (KA) is a neglected tropical disease caused by protozoan parasite, Leishmania sp. and is fatal, if left untreated. In this study, we measured trace elements (K, Fe, Cu, Zn, Br, Cl, S, Ca, Mn, Cr, Ni, As, Se, Rb and Sr) in the blood of Indian VL patients (32) by particle-induced X-ray emission (PIXE) study. Blood was collected from 36 subjects including healthy controls from Rambagh Kala-azar Hospital, Muzaffarpur, Bihar, India. PIXE experiment was carried out at the Institute of Physics, Bhubaneswar, India and data were analyzed by GUPIXWIN software. We observed first time the association of bromine with the disease. The results showed 48.47 % decrease in Br, 35.16 % decrease in Zn and 29.05 % decrease in Fe in untreated state of the KA patients. In the same group, Cu has been increased by 16.73 %. Cu/Zn ratio has been altered in diseased state. The association of bromine with the disease is reported for the first time and altered levels of trace elements (Br, Cu, Fe and Zn) may come back to normal after completion of the treatment regimen with Amphotericin B.

Vertical and horizontal variation of elemental contamination in sediments of Hooghly Estuary, India.

Hooghly Estuary along with the Sunderban mangroves forms one of the most diverse ecosystems in the world. We investigated the vertical and horizontal distribution of elements at nine sampling locations in this estuary for assessing the degree of elemental contamination. The elemental concentrations were analyzed by an Energy Dispersive X-ray Flurosence spectrometer (EDXRF). A higher enrichment factor (EF) value of trace elements (V, Cr, Mn, Fe, Ni, Cu and Zn) is evident considering all the sampling locations. Geo-accumulation index (Igeo) values of all the sampling locations and core depth reveals Cr and Cu are under Igeo Class I level of contamination. The value of the pollution load index (PLI) varies between 0.94-1.65 with maximum at Chemaguri and minimum at Haldi Estuary and Petuaghat. The overall variation in elemental concentration may be due to differential discharge pattern of storm water and agricultural run-off, industrial effluent and domestic sewage.

Effect of iron stress on Withania somnifera L.: antioxidant enzyme response and nutrient elemental uptake of in vitro grown plants.

In the present study the response of antioxidant enzyme activities and the level of expression of their corresponding genes on bioaccumulation of iron (Fe) were investigated. In vitro germinated Withania somnifera L. were grown in Murashige and Skoog's liquid medium with increasing concentrations (0, 25, 50, 100 and 200 µM) of FeSO4 for 7 and 14 days. Root and leaf tissues analyzed for catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (GPX, EC 1.11.1.7), have shown an increase in content with respect to exposure time. Isoforms of CAT, SOD and GPX were separated using non-denaturing polyacrylamide gel electrophoresis and observed that the isoenzymes were greatly affected by higher concentrations of Fe. Reverse transcriptase polymerase chain reaction analysis performed by taking three pairs of genes of CAT (RsCat, Catalase1, Cat1) and SOD (SodCp, TaSOD1.2, MnSOD) to find out the differential expression of antioxidant genes under Fe excess. RsCat from CAT and MnSOD from SOD have exhibited high levels of gene expression under Fe stress, which was consistent with the changes of the activity assayed in solution after 7 days of treatment. Analysis by proton induced X-ray emission exhibited an increasing uptake of Fe in plants by suppressing and expressing of other nutrient elements. The results of the present study suggest that higher concentration of Fe causes disturbance in nutrient balance and induces oxidative stress in plant.

Investigation on mechanism of Cr(VI) reduction and removal by Bacillus amyloliquefaciens, a novel chromate tolerant bacterium isolated from chromite mine soil.

A strain CSB 9 isolated from chromite mine soil of Sukinda, India was identified as Bacillus amyloliquefaciens based on biochemical and 16S rRNA gene sequencing. The strain exhibited relatively high tolerance to Cr(VI) (⩽900mgL(-1)) and fast reduction rate of 2.22mg Cr(VI) L(-1)h(-1), under optimized conditions of 100mgL(-1) Cr(VI), pH 7 and temperature 35°C within 45h. Mechanism of Cr(VI) reduction as well as nature and fate of the reduced product were studied to determine the scope of removal of reduced Cr(III) end product. AAS analyses of the culture products treated with Cr(VI) for 45h showed the distribution of Cr(III) in pellet and culture supernatant to be 37.4±1.7 and 62.6±3.4mgL(-1), respectively. In SEM images, the bacterial pellets with Cr(VI) treatment appeared coagulated, rough and porous whereas the pellets without Cr(VI) treatment appeared regular, smooth and non-porous in structure. SEM-EDX of the bacterial precipitates under Cr(VI) treatment revealed immobilization of Cr(III) species on the bacterial cell surface. Further Raman spectroscopy analysis confirmed the presence of Cr(III) species, with characteristic peak at around 600cm(-1). TEM-EDX study of the bacterial precipitates under Cr(VI) treatment showed intracellular deposition of Cr(III) which are in nanometric range. Further characterization of reduced product by XRD, FT-IR and SAED analyses suggested the formation of poorly crystalline end products. A Cr(VI) removal mechanism considering both the surface immobilization and intracellular accumulation of Cr(III) along with the formation of coagulated cell precipitate by living B. amyloliquefaciens was suggested.

Copper-stress induced alterations in protein profile and antioxidant enzymes activities in the in vitro grown Withania somnifera L.

Withania somnifera L. seedlings were grown in half-strength MS (Murashige and Skoog) basal medium for 4 weeks and then transferred to full-strength MS liquid medium for 3 weeks. The sustainable plants were subcultured in the same medium but with different concentrations (0, 25, 50, 100 and 200 µM) of Cu for 7 and 14 days. The growth parameters (root length, shoot length, leaf length and total number of leaves per plant) showed a declining trend in the treated plants in a concentration dependant manner. Roots and leaves were analyzed for protein profiling and antioxidant enzymes [catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (GPX, EC 1.11.1.7)]. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of crude protein extracts showed the appearance of some new proteins due to Cu treatment. In plant samples grown with 25 and 50 µM of Cu, a rapid increase in antioxidant activities were noticed but at higher concentration (100 and 200 µM) the activities declined. Isoforms of CAT, SOD and GPX were separated using non-denaturing polyacrylamide gel electrophoresis and concentration specific new isoforms were noticed during the study. Isoforms of the antioxidant enzymes synthesized due to Cu stress may be used as biomarkers for other species grown under metal stress.

Elemental alteration, iron overloading and metallothionein induction in experimental hepatocarcinogenesis: a free radical-mediated process?

The azo-dye p-dimethylaminoazobenzene (p-DAB) is a potential tumor initiator in rodents, but the underlying mechanism is not clear. Following chronic feeding of the carcinogen for 3, 5 and 7 weeks, trace elemental status, free radical generation, oxidative damage, antioxidant profile were measured in male and female swiss albino mice. The feeding resulted in iron accumulation in male mice liver. No increase in iron level was observed in similarly exposed female mice. The results of this study suggest that p-DAB-induced iron accumulation in male mice with concomitant production of oxidative free radicals is an early event in the hepatocarcinogenic initiation. This occurs selectively in male mice and affects either directly or indirectly in development of chemically induced liver neoplasia. Again, that upregulation of metallothionein (MT) expression in association with increased free radical generation was demonstrated in male mice. Alteration of copper (Cu) and zinc (Zn) levels are described in the light of antioxidant profile in liver tissue. The current results thus provide evidence in support of iron accumulations producing oxidative damage, and enhanced metallothionein expression as possible contributors in the mode of action of p-DAB induced hepatocarcinogenesis.

PIXE analysis of trace elements in relation to chlorophyll concentration in Plantago ovata Forsk.

Plantago ovata Forsk - an economically important medicinal plant - was analyzed for trace elements and chlorophyll in a study of the effects of gamma radiation on physiological responses of the seedlings. Proton-induced X-ray emission (PIXE) technique was used to quantify trace elements in unirradiated and gamma-irradiated plants at the seedling stage. The experiments revealed radiation-induced changes in the trace element and chlorophyll concentrations.

Analysis of trace elements during different developmental stages of somatic embryogenesis in Plantago ovata Forssk using energy dispersive X-ray fluorescence.

Energy dispersive X-ray fluorescence (ED-XRF) technique has been used for the determination of trace element profile during different developmental stages of somatic embryogenic callus of an economically important medicinal plant, Plantago ovata Forssk. Somatic embryogenesis is a plant tissue culture-based technique, which is used for plant regeneration and crop improvement. In the present investigation, elemental content was analysed using ED-XRF technique during different developmental stages and also determine the effect of additives--casein hydrolysate and coconut water on the trace elemental profile of embryogenic callus tissue of P. ovata. Subsequent experiments showed significant alteration in the concentration of K, Ca, Mn, Fe, Zn, Cu, Br, and Sr in both the embryogenic and non-embryogenic callus. Higher K, Ca, Fe, Cu, and Zn accumulation was in embryogenic tissue stage compared to other stages, suggesting these elements are crucial for successful embryogenesis. The results suggest that this information could be useful for formulating a media for in vitro embryo induction of P. ovata.

In vitro radiation induced alterations in heavy metals and metallothionein content in Plantago ovata Forsk.

Proton Induced X-ray emission (PIXE) and fluorescence-activated cell sorting (FACS) have been used to study the effects of gamma irradiation on heavy metal accumulation in callus tissue of Plantago ovata-an important cash crop of India. PIXE analysis revealed radiation-induced alteration in trace element profile during developmental stages of the callus of P. ovata. Subsequent experiments showed antagonism between Fe and Cu and also Cu and Zn and synergistic effect between Fe and Zn. FACS analysis showed significant induction of the metallothionein (MT) protein following gamma-irradiation, and maximum induction was noted at the 50-Gy absorbed dose. This indicated a progressive increment of MTs as a measure for protection against gamma-rays, to combat alteration in the homeostasis of heavy metals like Fe, Cu, Zn, and Mn.

Potential of Vinca rosea extracts in modulating trace element profile: a chemopreventive approach.

Diethylnitrosamine (DEN) was used as cancer-inducing agent in the experimental animals. Vinca rosea extract was supplemented with the drinking water as a chemopreventive agent. After 4 wk of treatment, animals were sacrificed and livers were excised. Nuclei and mitochondria were separated by differential centrifugation. The proton-induced X-ray emission technique has been used as the analytical method. Elemental analysis were performed for whole liver, nuclei, and mitochondria.V. rosea plant parts were also analyzed for elemental contents. Treatment with DEN caused an increase of Ni, Zn, and Cr levels in the whole liver and nuclei. There is an increase in Fe concentration in the liver, although the level decreased in mitochondria. The concentrations of Br and Ca were unchanged in the liver as a whole, but there were substantial increases of Br in nuclei and mitochondria, whereas Ca levels depleted drastically in these two organelles. Vinca extracts were effective in reverting the changes in the elemental concentration in the hepatic tissue as a whole, but were not that effective at subcellular levels.