Purification and characterization of a bacteriocin-like compound (Lichenin) produced anaerobically by Bacillus licheniformis isolated from water buffalo.

AIMS: To characterize a bacteriocin-like factor from Bacillus licheniformis 26 L-10/3RA isolated from buffalo rumen. METHODS AND RESULTS: The culture supernatant exhibited the antibacterial activity against a number of indicator organisms in a cut-well agar assay under anaerobic conditions. The inhibitory component was purified by following ammonium sulphate precipitation, gel filtration and ion exchange chromatography and confirmed to be a single peptide. A single band on tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis confirmed that the peptide was purified to homogeneity and having an estimated molecular mass of approximately 1400 dalton. Complete amino acid sequence of the peptide yielded 12 amino acids from the N-terminal end (ISLEICXIFHDN). No homology with previously reported bacteriocins was observed and has been designated as Lichenin. Lichenin was found to be hydrophobic, sensitive to atmospheric oxygen, retained biological activity even after boiling for 10 min and was active over a pH range of 4.0-9.0. CONCLUSIONS: The Lichenin represents the first anaerobiosis specific expression of bacteriocin-like compound isolated from Bacillus licheniformis 26 L-10/3RA of buffalo rumen origin. SIGNIFICANCE AND IMPACT OF THE STUDY: Lichenin could be a potential candidate for manipulating the rumen function at molecular level intended for improving the productivity of the ruminant.

Edwardsiella tarda endotoxin as an immunopotentiator in Singhi, Heteropneustes fossilis fingerlings.

Endotoxin of E. tarda grown in brain heart infusion broth at 30 degrees C for 18 hr was extracted by differential centrifugation. Fingerlings of H. fossilis (weighing 1-2 g) were allowed for hyperosmotic infiltration in the endotoxin at the rate of 0,2,4,8,16 and 20 mg/l. Mortality varied from 20-50% at 2 to 20 mg/ml. Toxin treated fishes were challenged 21 days post treatment with the same E. tarda strain containing 2.1 x 10(9) CFU/ml. There was 80% mortality in the control group whereas only 20% mortality in toxin treated group at 2 mg/l concentration after challenge with homologous E. tarda. Subsequently a second challenge of E. tarda was given to the survivors of fish one month after first challenge using same concentration where no mortality could be observed. It was concluded that the endotoxin could enhance percentage of survival against E. tarda infection in Singhi.

Influence of salinity on methanogenesis and associated microflora in tropical rice soils.

In a laboratory incubation study, methane (CH4) production in two saline soils and a nonsaline soil sample was investigated under flooded conditions. Mean CH4 production was remarkable (630.86 ng CH4/g) in nonsaline alluvial soil, but low (12.97 ng CH4/g) in acid sulfate saline (Pokkali) soil which was attributed to the high sulfate content of the later. CH4 production was also low in the coastal saline (Canning) soil (142.36 ng CH4/g) but increased upon leaching the soil of its salt content. Addition of salts to the nonsaline alluvial soil at 4, 8, 16 and 20 dS/m progressively decreased CH4 production. The inhibition of CH4 production was related to low microbial activities as reflected by decreased microbial biomass C and low soil microbial population including that of methanogens.

Two possible conducting states of the influenza A virus M2 ion channel.

Molecular dynamics simulations have been performed on protonated four-helix bundles based on the 25-residue Duff-Ashley transmembrane sequence of the M2 channel of the influenza A virus. Well-equilibrated tetrameric channels, with one, two and four of the H37 residues protonated, were investigated. The protonated peptide bundles were immersed in the octane portion of a phase-separated water/octane system, which provided a membrane-mimetic environment. The simulations suggest that there could be two conducting states of the M2 channel corresponding to tetramers containing one or two protonated histidines. The more open structure of the doubly protonated state suggests it would have the higher conductance.

The M2 channel of influenza A virus: a molecular dynamics study.

Molecular dynamics simulations have been performed on a tetramer of the 25-residue (SSDPLVVAASIIGILHLILWILDRL) synthetic peptide [1] which contains the transmembrane domain of the influenza A virus M2 coat protein. The peptide bundle was initially assembled as a parallel alpha-helix bundle in the octane portion of a phase separated water/octane system, which provided a membrane-mimetic environment. A 4-ns dynamics trajectory identified a left-handed coiled coil state of the neutral bundle, with a water filled funnel-like structural motif at the N-terminus involving the long hydrophobic sequence. The neck of the funnel begins at V27 and terminates at H37, which blocks the channel. The C-terminus is held together by inter-helix hydrogen bonds and contains water below H37. Solvation of the S23 and D24 residues, located at the rim of the funnel, appears to be important for stability of the structure. The calculated average tilt of the helices in the neutral bundle is 27 +/- 5 degrees, which agrees well with recent NMR data.

Molecular dynamics simulation of a hydrated diphytanol phosphatidylcholine lipid bilayer containing an alpha-helical bundle of four transmembrane domains of the influenza A virus M2 protein.

An alpha-helical bundle composed of four transmembrane portions of the M2 protein from the Influenza A virus has been studied in a hydrated diphytanol phosphatidylcholine bilayer using molecular dynamics (MD) calculations. Experimentally, the sequence utilized is known to aggregate as a four-helix bundle and act as a pH-gated proton-selective ion channel, which is blocked by the drug amantadine hydrochloride. In the presented simulation, the ion channel was initially set up as a parallel four-helix bundle. The all-atom simulation consisted of almost 16,000 atoms, described classically, using a forcefield from the CHARMM22 database. Bilayers with and without the bundle were shown to be stable throughout the nanosecond timescale of the MD simulation. Structural and dynamical properties of the bilayer both with and without the transmembrane protein are reported.

Knotted upper end: a new complication in the use of an indwelling ureteral stent.

Use of the indwelling ureteral stent is a key factor in the management of stone disease by extracorporeal shock wave lithotripsy. We report on a knotted stent in a patient with a solitary kidney, which had to be removed percutaneously. This case demonstrates that the ureteral stent is not free of serious complications.