Bioproduction and characterization of extracellular melanin-like pigment from industrially polluted metagenomic library equipped Escherichia coli.

To explore the potential genes from the industrially polluted Amlakhadi canal, located in Ankleshwar, Gujarat, India, its community genome was extracted and cloned into E. coli EPI300™-T1R using a fosmid vector (pCC2 FOS™) generating a library of 3,92,000 clones with average size of 40kb of DNA-insert. From this library, the clone DM1 producing brown colored melanin-like pigment was isolated and characterized. For over expression of the pigment, further sub-cloning of the clone DM1 was done. Sub-clone containing 10kb of the insert was sequenced for gene identification. The amino acids sequence of a protein 4-Hydroxyphenylpyruvate dioxygenase (HPPD), which is know to be involved in melanin biosynthesis was obtained from the gene sequence. The sequence-homology based 3D structure model of HPPD was constructed and analyzed. The physico-chemical nature of pigment was further analysed using 1H and 13C NMR, LC-MS, FTIR and UV-visible spectroscopy. The pigment was readily soluble in DMSO with an absorption maximum around 290nm. Based on the genetic and chemical characterization, the compound was confirmed as melanin-like pigment. The present results indicate that the metagenomic library from industrially polluted environment generated a microbial tool for the production of melanin-like pigment.

Benzothiazole analogues: Synthesis, characterization, MO calculations with PM6 and DFT, in silico studies and in vitro antimalarial as DHFR inhibitors and antimicrobial activities.

Benzothiazole analogues are of interest due to their potential activity against malarial and microbial infections. In search of suitable antimicrobial and antimalarial agents, we report here the synthesis, characterization and biological activities of benzothiazole analogues (J 1-J 10). The molecules were characterized by IR, Mass, 1H NMR, 13C NMR and elemental analysis. The in vitro antimicrobial activity was investigated against pathogenic strains; the results were explained with the help of DFT and PM6 molecular orbital calculations. In vitro cytotoxicity and genotoxicity of the molecules were studied against S. pombe cells. In vitro antimalarial activity was studied. The active compounds J 1, J 2, J 3, J 5 and J 6 were further evaluated for enzyme inhibition efficacy against the receptor Pf-DHFR, computational and in vitro studies were carried out to examine their candidatures as lead dihydrofolate reductase inhibitors.

Phytol induces ROS mediated apoptosis by induction of caspase 9 and 3 through activation of TRAIL, FAS and TNF receptors and inhibits tumor progression factor Glucose 6 phosphate dehydrogenase in lung carcinoma cell line (A549).

A number of drugs as well as lead molecules are isolated from natural sources. Phytol is one of such lead molecule belongs to terpenes group distributed widely in medicinal plants. In the present work, we investigated the cytotoxic behavior of phytol on human lung carcinoma cells (A549). Phytol was found to cause characteristic apoptotic morphological changes and generation of ROS in A549 cells. The mechanism of phytol involved the activation of TRAIL, FAS and TNF-α receptors along with caspase 9 and 3. In silico molecular docking studies revealed that phytol has a good binding affinity with glucose-6-phosphate dehydrogenase (G6PD), which is known to promote tumor proliferation. The ability of phytol to become potential drug candidate has been revealed from the pharmacokinetic study performed in the present study.

1,2,4-Triazole and 1,3,4-oxadiazole analogues: Synthesis, MO studies, in silico molecular docking studies, antimalarial as DHFR inhibitor and antimicrobial activities.

1,2,4-Triazole and 1,3,4-oxadiazole analogues are of interest due to their potential activity against microbial and malarial infections. In search of suitable antimicrobial and antimalarial compounds, we report here the synthesis, characterization and biological activities of 1,2,4-triazole and 1,3,4-oxadiazole analogues (SS 1-SS 10). The molecules were characterized by IR, mass, 1H NMR, 13C NMR and elemental analysis. The in vitro antimicrobial activity was investigated against pathogenic strains, the results were explained with the help of DFT and PM6 molecular orbital calculations. In vitro cytotoxicity and genotoxicity of the molecules were studied against S. pombe cells. In vitro antimalarial activity was studied. The active compounds were further evaluated for enzyme inhibition efficacy against the receptor Pf-DHFR computationally as well as in vitro to prove their candidature as lead dihydrofolate reductase inhibitors.

Nano silver-embedded electrospun nanofiber of poly(4-chloro-3-methylphenyl methacrylate): use as water sanitizer.

Water contaminated with microorganisms causes numerous diseases and is a major concern for public health. In search of a simple material which can provide clean water free from pathogens, nanofibers of poly(4-chloro-3-methylphenyl methacrylate, abbreviated as CMPMA, and nano Ag-doped poly(CMPMA) composite nanofibers were used to decontaminate water from microorganisms such as Escherichia coli and Bacillus subtilis. Nanofibers were prepared by electrospinning. X-ray diffraction (XRD) and transmission electron microscopy (TEM) provide the diameters of the Ag nanoparticles which are in the range 18-21 and 13-18 nm. The diameter of the poly(CMPMA) and nano Ag-doped poly(CMPMA) composite nanofiber is seen to vary between 400 and 700 nm with the change of the processing parameters. Optimum parameters for uniform nanofibers have been obtained. The morphology of the fibers is derived from scanning electron microscopy (SEM). The superiority of the nano Ag-doped poly(CMPMA) composite nanofiber was established.

Synthesis and biological screening of novel 2-morpholinoquinoline nucleus clubbed with 1,2,4-oxadiazole motifs.

Novel series of 2-morpholinoquinoline scaffolds (6a-n), containing the 1,2,4-oxadiazole and moiety, was designed and synthesized in good yield (76-86%). The synthesized compounds were screened for their preliminary in vitro antimicrobial activity against a panel of pathogenic strains of bacteria and fungi. Molecular docking and pharmacokinetic study were carried out for the prepared compounds. The cytotoxicity of the synthesized compounds was tested at different concentrations using bioassay of S. pombe cells at the cellular level. The effect of synthesized compounds on the DNA integrity of S. pombe was observed on agarose gel. Compounds 6d, 6e, 6g, 6h, 6j and 6n exhibited excellent antimicrobial potency as compared to the standard drugs (i.e Ampicillin, Norfloxacin, Chloramphenicol, Ciprofloxacin). Compounds 6d, 6e, 6g, 6k and 6n were found to have significant antifungal activity as compared to griseofulvin. Compounds 6f, 6i, 6k, 6l were found very less cytotoxic, while compounds 6d, 6e, 6g, 6h were found to exhibit maximum toxicity. The rest of the synthesized compounds were found to be moderately toxic.

Synthesis, characterization and in silico designing of diethyl-3-methyl-5-(6-methyl-2-thioxo-4-phenyl-1,2,3,4-tetrahydropyrimidine-5-carboxamido) thiophene-2,4-dicarboxylate derivative as anti-proliferative and anti-microbial agents.

A series of eight compounds diethyl-3-methyl-5-(6-methyl-2-thioxo-4-phenyl-1,2,3,4-tetrahydropyrimidine-5-carboxamido) thiophene-2,4-dicarboxilate (KM10-17) analogues have been prepared by conventional methods and characterized by IR, Mass, NMR and elemental analysis. In silico docking studies on Human topoisomerase IIbeta (PDB Id: 3QX3) have been performed for all molecules (KM10-17) synthesized. The compounds were tested for in vitro anti-proliferative activity on VERO and 786-O cell lines. Out of all the synthesized compounds, KM11 &KM16 showed moderate activity on both cell lines. In vitro anti-microbial activity was also checked against Bacillus subtilis (BS), Staphylococcus aurous (SA), Pseudomonas aeruginosa (PA), Escherichia coli (EC) and Candida albicans (CA) by well diffusion method. The compound KM11 was found to have highest zone of inhibition against BS, SA, PA and EC. The molecules KM13 and KM16 exhibited good activity against CA. The compounds KM14 and KM16 indicated good zone of inhibition against BS.

6-Tosyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide analogues: synthesis, characterization, MO calculation, and antibacterial activity.

A series of 6-tosyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide analogues are synthesized by conventional techniques and characterized by elemental analysis, IR, MS, (1)H, and (13)C NMR. These are tested for their antibacterial activity against Bacillus subtilis (abbreviated as BS), Staphylococcus aureus (abbreviated as SA), and Escherichia coli (abbreviated as EC). The synthesized compounds are able to inhibit the growth of the SA and EC. None of the compounds are effective against BS. All valence molecular orbital (abbreviated as MO) calculations with PM6 have been carried out for the molecules for which bioactivity data are available. Ciprofloxacin is taken as the standard antibiotics to compare activity with the molecules synthesized. It has been attempted to correlate the activity of the molecules with their electronic structure.

Study of pulmonary functions in tea industry workers in North Bengal.

This study was intended to compare pulmonary function parameters in tea garden factory workers who are on chronic exposure to tea dust with tea garden plantation workers on the basis of dynamic ventilatory pulmonary function tests. The pulmonary function tests done on 128 subjects included 64 tea garden factory workers and 64 tea garden plantation workers. In tea factory workers the values of vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in one second (FEV1), forced expiratory volume in first 0.75 second (FEV0.75), FEV1/FVC%, peak expiratory flow rate (PEFR) were found significantly lower (p < 0.001) than those of tea garden plantation workers. It may be concluded that pulmonary functions are more affected in tea factory workers than those of tea plantation workers.

Assessment of iodine and non-iodine deficiency hypothyroidism in women of reproductive ages in the sub-Himalayan plains of West Bengal.

Indian women of reproductive age groups commonly suffer from hypothyroidism which may be due to iodine or non iodine deficiency causes. This study was undertaken with a view to ascertain the leading cause of hypothyroidism in women of reproductive age group residing in the sub-Himalayan plain areas of Darjeeling district of West Bengal. Serum TSH, T4, T3 and Urinary Iodine Excretion (UIE) levels were measured in 101 non pregnant women. Our results reveal that among 37.62% (n=38) of bio-chemically established hypothyroid women; majority 76.32% (n=29) are suffering from iodine deficiency and the rest 23.68% (n=9) have Hypothyroidism due to other causes. Moreover, iodine deficiency persists among 57.42% (n=58) of the women in our study. We conclude that iodine deficiency disorders are still a major problem in this region and hypothyroidism due to iodine deficiency is more prevalent than the non iodine deficiency causes. Hence lacunae in the iodine supplementation process needs to be reviewed.

Bioaccumulation of heavy metals by green algae.

The biosorption of metal ions (Cr(+3), Cr(2)O(7)(-2), Cu(+2), and Ni(+2)) on two algal blooms (designated HD-103 and HD-104) collected locally was investigated as a function of the initial metal ion concentration. The main constituent of HD-103 is Cladophora sp., while Spirulina sp. is present significantly in the bloom HD-104. Algal biomass HD-103 exhibited the highest Cu(+2) uptake capacity (819 mg/g). This bloom adsorbed Ni(+2) (504 mg/g), Cr(+3) (347 mg/g), and Cr(2)O(7)(-2), (168 mg/g). Maximum of Ni(+2) (1108 mg/g) is taken by HD-104. This species takes up 306, 202, and 576 mg/g Cr(+3), Cr(2)O(7)(-2), and Cu(+2), respectively. Equilibrium data fit very well to both the Langmuir and the Freundlich isotherm models. The sorption process followed the Freundlich model better. Pseudo-first-order kinetic model could describe the kinetic data. Infrared (IR) spectroscopic data were employed to identify the site(s) of bonding. It was found that phosphate and peptide moieties participate in the metal uptake by bloom HD-103. In the case of bloom HD-104, carboxylate and phosphate are responsible for the metal uptake. The role of protein in metal uptake by HD-103 was investigated using polyacrylamide gel electrophoresis.

Bioremediation potential of Chlorella: spectroscopic, kinetics, and SEM studies.

A dead dried alga, Chlorella sp., was used for the uptake of Cr+3, Cr2O7(-2), Cu+2, and Ni+2 from the aqueous solutions of these metal ions. The equilibrium data were fitted using the Langmuir and Freundlich isotherm model and the maximum uptakes for Cr+3, Cr2O7(-2), Ni+2, and Cu+2 were 98, 104, 108, and 183 mg/g, respectively. The Freundlich model, in comparison to the Langmuir model, better represented the sorption process. The kinetics of metal ions uptake by Chlorella sp. was best described by a pseudo-second order rate equation. Infrared spectroscopic data were employed to identify the site(s) of bonding in Chlorella sp. A scanning electron microscopic (SEM) study of pure dead Chlorella sp. and the species treated with different metal ions provided an idea of the extent of metal uptake by this species. The dead Chlorella sp took up maximum Cu(II). The size of the cell of the metal-treated Chlorella sp. obtained from SEM data is in agreement with the extent of metal uptake.

Biosorption of cadmium by live and dead Spirulina: IR spectroscopic, kinetics, and SEM studies.

Cadmium is an important environmental pollutant and a potent toxicant to bacteria, algae, and fungi. Mechanisms of Cd+2 toxicity and resistance are variable, depending on the organism. The present work reports the use of live and dead Spirulina sp. for sorption of Cd+2. This investigation shows that this biomass takes up substantial amount of Cd+2 ions. IR spectroscopic study, kinetics models, Langmuir & Freundlich adsorption isotherms, scanning electron microscopic analysis of Spirulina sp., and the Spirulina sp. treated with different metal ions have been employed to understand the sorption mechanism. Infrared spectra of live Spirulina treated with Cd+2 ions for different lengths of time have been taken to understand the time dependency of metal interaction.

Bioremediation potential of live and dead Spirulina: spectroscopic, kinetics and SEM studies.

Metal binding by algae has enormous potential for environmental bioremediation targeting towards detoxification of water bodies. The present work reports the use of live and dead Spirulina sp. for sorption of metals like Cr(3+), Ni(2+), Cu(2+), and Cr(6+) in form of Cr(2)O(7)(2-). Preliminary investigation shows that this biomass takes up substantial amount of metal ions indicated above. IR spectroscopic study, kinetics models, Langmuir and Freundlich adsorption isotherms, SEM analysis, and fluorescence microscopic study of Spirulina sp. and the Spirulina sp. treated with different metal ions have been employed to understand the sorption mechanism. It is hoped that live Spirulina sp. will be a strong candidate for management of industrial wastewater.

Spectroscopic and scanning electron microscopy studies of bioaccumulation of pollutants by algae.

The ability of Chlorella species and two other algal blooms collected locally to take up Cu(+2) and Ni(+2) was investigated using infrared and scanning electron microscopy (SEM) data. The percentage of metal uptake was determined with atomic absorption spectroscopy. The effects of pH and initial concentrations of metal ions on bioaccumulation were examined. The uptake of methyl orange dye by Chlorella species was also studied using Langmuir and Freundlich adsorption isotherms.