The contribution of endophytic bacteria to Albizia lebbeck-mediated phytoremediation of tannery effluent contaminated soil

Toxicity of chromium often impairs the remediation capacity of plants used in phytoremediation of polluted soils. In this study, we have identified Albizia lebbeck as a prospective chromium hyperaccumulator and examined cultivable diversity of endophytes present in chromium-treated and control saplings. High numbers (22–100%) of endophytic bacteria, isolated from root, stem, and leaf tissues, could tolerate elevated (1–3 mM) concentrations of K₂CrO₇. 16S rRNA gene sequence-based phylogenetic analysis showed that the 118 isolates obtained comprised of 17 operational taxonomic units affiliated with the proteobacterial genera Rhizobium (18%), Marinomonas (1%), Pseudomonas (16%), and Xanthomonas (7%) but also with members of Firmicutes genera, such as Bacillus (35%) and Salinococcus (3%). The novel isolates belonging to Salinococcus and Bacillus could tolerate high K₂CrO₇ concentrations (3 mM) and also showed elevated activity of chromate reductase. In addition, majority (%) of the endophytic isolates also showed production of indole-3-acetic acid. Taken together, our results indicate that the innate endophytic bacterial community assists plants in reducing heavy metal toxicity.     

Environmental Risk Assessment Studies of Heavy Metal Contamination in the Industrial Area of Kattedan,India—A Case Study

Kattedan is an industrial area near Hyderabad, Andhra Pradesh, India, contaminated with high concentrations of metals attributed to industrial sources (battery manufacturing, metal plating, textile and pharmaceuticals production and others). Twelve different locations in the Kattedan industrial area were assessed for concentrations of metals (Zn, Cr, Cu, Ni, Co, Pb, Hg, Cd, and As) in soils, waters, and vegetation. Application of sequential extraction technique for the soils revealed relatively high percentages of Zn, Cu, and Cr associated with mobile fractions, and correspondingly high concentrations of Zn, Cr, Cu, and Pb in forage grass samples and a high degree of bioavailability to humans. Human exposure assessment revealed high concentrations of Pb, Zn, and Cr in blood and urine samples from the residents of the study area showing a direct pathway and a potential for toxicological hazard due to heavy metal pollution.     

Isolation,structural identification and herbicidal activity of N-phenylpropanamide from Streptomyces sp. KA1-3

The secondary metabolites produced by Streptomyces sp. KA₁-3, cultured on starch casein broth, was extracted by ethyl acetate and concentrated. Purification of the compound by thin layer chromatography lead to isolation of N-phenylpropanamide from one polar fraction. The structure of the herbicidal compound was elucidated on the basis of UV, FT-IR, mass and H¹ NMR spectroscopy. The herbicidal activity of the isolate was tested against the weeds Cassia occidentalis L. and rhizome Cyperus rotundus L. by moist chamber technique and rolled towel paper assay method. Herbicidal activity of the bioactive compound N-phenylpropanamide was further evaluated under in vitro condition. The herbicidal compound showed 80% of seed germination inhibition in C. occidentalis L. and rhizome C. rotundus L. weed. The actinobacterium can be used as a source for bioherbicidal agent.     

Isolation and partial characterization of collagen from outer skin of Sepia pharaonis (Ehrenberg, 1831) from Puducherry coast

Type I collagen from outer skin of Sepia pharaonis was extracted and partially characterized. Yield of Acid Soluble Collagen (ASC) and Pepsin Soluble Collagen (PSC) were calculated as 1.66% and 3.93% and the total protein content of ASC and PSC were found as 18.4% and 48.6%. FT-IR spectrum of ASC and PSC recorded 12 and 14 peaks, respectively. ¹H NMR spectrum of ASC showed singlets at 1.23 ppm, 3.1 ppm, 3.55 ppm and 3.7 ppm and PSC at 1.23 ppm and 2.08 ppm. The molecular weight for ASC was calculated as 102 kDa and for PSC as 110, 108 and 102 kDa through SDS-PAGE. Differential Scanning Calorimetry (DSC) results supported that PSC withstand high thermal stability (82.85 °C) than ASC (73.13 °C). Higher denaturation temperature with high molecular weight well support the property of type I collagen from skin of S. pharaonis and it could be used as another potent source for the extraction of collagen.     

Critical role of bicarbonate and bicarbonate transporters in cardiac function

Bicarbonate is one of the major anions in mammalian tissues and extracellular fluids. Along with accompanying H+, HCO3- is generated from CO2 and H2 O, either spontaneously or via the catalytic activity of carbonic anhydrase. It serves as a component of the major buffer system, thereby playing a critical role in pH homeostasis. Bicarbonate can also be utilized by a variety of ion transporters, often working in coupled systems, to transport other ions and organic substrates across cell membranes. The functions of HCO3- and HCO3--transporters in epithelial tissues have been studied extensively, but their functions in heart are less well understood. Here we review studies of the identities and physiological functions of Cl-/HCO3- exchangers and Na+/HCO3-cotransporters of the SLC4 A and SLC26 A families in heart. We also present RNA Seq analysis of their cardiac mRNA expression levels. These studies indicate that slc4a3(AE3) is the major Cl-/HCO3- exchanger and plays a protective role in heart failure, and that Slc4a4(NBCe1) is the major Na+/HCO3- cotransporter and affects action potential duration. In addition, previous studies show that HCO3- has a positive inotropic effect in the perfused heart that is largely independent of effects on intracellular Ca2+. The importance of HCO3- in the regulation of contractility is supported by experiments showing that isolated cardiomyocytes exhibit sharply enhanced contractility, with no change in Ca2+ transients, when switched from Hepes-buffered to HCO3-- buffered solutions. These studies demonstrate that HCO3- and HCO3--handling proteins play important roles in the regulation of cardiac function.     

Effect of H4R antagonist N-(2-aminoethyl)-5-chloro-1H-indol-2-carboxamides and 5-chloro-2-(piperazin-1-ylmethyl)-1H-benzimidazole on histamine and 4-methylhistamine-induced mast cell response

Context: The histamine plays a decisive role in acute and chronic inflammatory responses and is regulated through its four types of distinct receptors designated from H1 to H4. Recently histamine 4 receptor (H4R) antagonists have been reported to possess various pharmacological effects against various allergic diseases.

Objective: To investigate the inhibitory effect of N-(2-aminoethyl)-5-chloro-1H-indol-2-carboxamide (Compound A) and 5-chloro-2-(piperazin-1-ylmethyl)-1H-benzimidazole (Compound L) on H4R-mediated calcium mobilization, cytokine IL-13 production, ERK1/2, Akt and NF-κB activation in human mastocytoma cells-1 (HMC-1).

Materials and methods: Compounds A and L were synthesized chemically and their inhibitory effect on intracellular calcium release was analyzed by Fluo-4 calcium assay, cytokine measurement through ELISA and activation of signaling molecules by western blot.

Results: Pre-treatment with compounds A and L significantly reduced the H4R-mediated intracellular calcium release. Histamine and 4-methylhistamine (4-MH) induced Th2 cytokine IL-13 production in HMC-1 cells, was inhibited by compound A (77.61%, 74.25% at 1?μM concentration) and compound L (79.63%, 81.70% at 1?μM concentration). Furthermore, histamine induced the phosphorylation of ERK1/2, Akt and NF-κB was suppressed by compounds A and L at varying levels, ERK1/2 (88%, 86%), Akt (88%, 89%) and NF-κB (89%, 87%) in HMC-1 cells.

Discussion and conclusions: Taken together these data demonstrate that compound A and compound L may block H4R-mediated downstream signaling events.     

Channel-forming, self-assembling, bishelical amphiphilic peptides: design, synthesis and crystal structure of Py(Aibn)2, n=2,3,4.

The design, synthesis, characterization and self-assembling properties of a new class of amphiphilic peptides, constructed from a bifunctional polar core attached to totally hydrophobic arms, are presented. The first series of this class, represented by the general structure Py(Aibn)2 (Py=2,6-pyridine dicarbonyl unit; Aib=alpha, alpha'-dimethyl glycine; n=1-4), is prepared in a single step by the condensation of commercially available 2,6-pyridine dicarbonyl dichloride with the methyl ester of homo oligoAib peptide (Aibn-OMe) in the presence of triethyl amine. 1H NMR VT and ROESY studies indicated the presence of a common structural feature of 2-fold symmetry and an NH...N hydrogen bond for all the members. Whereas the Aib3 segment in Py(Aib3)2 showed only the onset of a 3(10)-helical structure, the presence of a well-formed 3(10)-helix in both Aib4 arms of Py(Aib4)2 was evident in the 1H NMR of the bispeptide. X-ray crystallographic studies have shown that in the solid state, whereas Py(Aib2)2 molecules organize into a sheet-like structure and Py(Aib3)2 molecules form a double-stranded string assembly, the tetra Aib bispeptide, Py(Aib4)2, is organized to form a tetrameric assembly which in turn extends into a continuous channel-like structure. The channel is totally hydrophobic in the interior and can selectively encapsulate lipophilic ester (CH3COOR, R=C2H5, C5H11) molecules, as shown by the crystal structures of the encapsulating channel. The crystal structure parameters are: 1b, Py(Aib2)2, C25H37N5O8, sp. gr. P2(1)2(1)2(1), a=9.170(1) A, b=16.215(2) A, c=20.091(3) A, R=4.80; 1c, Py(Aib3)2, C33H51N7O10H2O, sp. gr. P1, a=11.040(1) A, b=12.367(1) A, c=16.959(1) A, alpha =102.41 degrees, beta =97.29 degrees, gamma =110.83 degrees, R1=6.94; 1 da, Py(Aib4)2.et ac, C41H65N9O12.1.5H2O.C4H8O2, sp. gr. P1, a=16.064(4) A, b=16.156 A, c=21.655(5) A, alpha =90.14(1)degrees, beta=101.38(2) degrees, gamma=97.07(1)degrees, Z=4, R1=9.03; 1db, Py(Aib4)2.amylac, C41H65N9O12.H2O.C7H14O2, P2(1)/c, a=16.890(1) A, b=17.523(1)A, c=20.411(1) A, beta=98.18 degrees, Z=4, R=11.1 (with disorder).     

<Emphasis Type="Italic">Planococcus antarcticus</Emphasis> and<Emphasis Type="Italic"> Planococcus psychrophilus</Emphasis> spp. nov. isolated from cyanobacterial mat samples collected from ponds in Antarctica

Thirteen orange-pigmented bacteria associated with cyanobacterial mat samples collected from four different lakes in McMurdo, Antarctica, were isolated. Twelve of the isolates, which were coccoid in shape, were very similar and possessed all the characteristics of the genus Planococcus and represented a new species, which was assigned the name Planococcus antarcticus sp. nov. (CMS 26or(T)). Apart from the phenotypic differences, P. antarcticus differed from all reported species of Planococcus by more than 2.5% at the 16S rRNA gene sequence level. In addition, at the DNA-DNA hybridization level, it exhibited very little similarity either with P. mcmeekinii (30%-35%), P. okeanokoites (26%-29%), or CMS 53or(T) (15%-25%), the three species with which it is closely related at the rRNA gene sequence level (2.5%-2.9%). P. antarcticus also showed only 2.5% difference in its 16S rRNA gene sequence compared with the P. alkanoclasticus sequence. But it was distinctly different from P. alkanoclasticus, which exists only as rods, is mesophilic and phosphatase positive, can hydrolyze starch, cannot utilize succinate, glutamate, or glucose, and cannot acidify glucose. Most important, P. antarcticus and P. alkanoclasticus varied distinctly in their fatty acid composition in that C(15:0), C(15:1), C(16:0), iso-C(16:1), and C(17:0) were present only in P. antarcticus but absent in P. alkanoclasticus. CMS 53or(T), the thirteenth isolate, was also identified as a new species of Planococcus and was assigned the name Planococcus psychrophilus sp. nov. This species was distinctly different from all the reported species, including the new species P. antarcticus, with respect to a number of phenotypic characteristics. At the 16S rRNA gene sequence level, it was closely related to P. okeanokoites (98.1%) and P. mcmeekinii (98%), but with respect to the DNA-DNA hybridization, the similarity was only 35%-36%. The type strain of P. antarcticus is CMS 26or(T) (MTCC 3854; DSM 14505), and that of P. psychrophilus is CMS 530r(T) (MTCC 3812; DSM 14507).     

Tigerinins: novel antimicrobial peptides from the Indian frog Rana tigerina

Four broad-spectrum, 11 and 12 residue, novel antimicrobial peptides have been isolated from the adrenaline-stimulated skin secretions of the Indian frog Rana tigerina. Sequences of these peptides have been determined by automated Edman degradation, by mass spectral analysis and confirmed by chemical synthesis. These peptides, which we have named as tigerinins, are characterized by an intramolecular disulfide bridge between two cysteine residues forming a nonapeptide ring. This feature is not found in other amphibian peptides. Conformational analysis indicate that the peptides tend to form beta-turn structures. The peptides are cationic and exert their activity by permeabilizing bacterial membranes. Tigerinins represent the smallest, nonhelical, cationic antimicrobial peptides from amphibians.