Dynamics imaging of lipid phases and lipid-marker interactions in model biomembranes

Biomembranes are complex systems that regulate numerous biological processes. Lipid phases that constitute these membranes influence their properties and transport characteristics. Here, we demonstrate the potential of short-range dynamics imaging (excited-state lifetime, rotational diffusion, and order parameter) as a sensitive probe of lipid phases in giant unilamellar vesicles (GUVs). Liquid-disordered and gel phases were labeled with Bodipy-PC at room temperature. Two-photon fluorescence lifetime imaging microscopy of single-phase GUVs reveals more heterogeneity in fluorescence lifetimes of Bodipy in the gel phase (DPPC: 3.8+/-0.6 ns) as compared with the fluid phase (DOPC: 5.2+/-0.2 ns). The phase-specificity of excited-state lifetime of Bodipy-PC is attributed to the stacking of ordered lipid molecules that possibly enhances homo-FRET. Fluorescence polarization anisotropy imaging also reveals distinctive molecular order that is phase specific. The results are compared with DiI-C12-labeled fluid GUVs to investigate the sensitivity of our fluorescence dynamics assay to different lipid-marker interactions. Our results provide a molecular perspective of lipid phase dynamics and the nature of their microenvironments that will ultimately help our understanding of the structure-function relationship of biomembranes in vivo. Furthermore, these ultrafast excited-state dynamics will be used for molecular dynamics simulation of lipid-lipid, lipid-marker and lipid-protein interactions.     

Salt-tolerant and thermostable alkaline protease fromBacillus subtilis NCIM No. 64

The proteolytic activity produced by aBacillus subtilis isolated from a hot spring was investigated. Maximum protease production was obtained after 38 h of fermentation. Effects of various carbon and nitrogen sources indicate the requirement of starch and bacteriological peptone to be the best inducers for maximum protease production. Requirement for phosphorus was very evident, and the protease was secreted over a wide range of pH 5–11. The partially purified enzyme was stable at 60°C for 30 min. Calcium ions were effective in stabilizing the enzyme, especially at higher temperature. The enzyme was extremely salt tolerant and retained 100% activity in 5M NaCl over 96 h. The molecular weight of the purified enzymes as determined by SDS-PAGE was 28,000. The enzyme was completely inactivated by PMSF, but little affected by urea, sodium dodecyl sulfate, and sodium tripoly phosphate.     

Calcium-Dependent Translocation of S100B Is Facilitated by Neurocalcin Delta

S100B is a calcium-binding protein that governs calcium-mediated responses in a variety of cells—especially neuronal and glial cells. It is also extensively investigated as a potential biomarker for several disease conditions, especially neurodegenerative ones. In order to establish S100B as a viable pharmaceutical target, it is critical to understand its mechanistic role in signaling pathways and its interacting partners. In this report, we provide evidence to support a calcium-regulated interaction between S100B and the neuronal calcium sensor protein, neurocalcin delta both in vitro and in living cells. Membrane overlay assays were used to test the interaction between purified proteins in vitro and bimolecular fluorescence complementation assays, for interactions in living cells. Added calcium is essential for interaction in vitro; however, in living cells, calcium elevation causes translocation of the NCALD-S100B complex to the membrane-rich, perinuclear trans-Golgi network in COS7 cells, suggesting that the response is independent of specialized structures/molecules found in neuronal/glial cells. Similar results are also observed with hippocalcin, a closely related paralog; however, the interaction appears less robust in vitro. The N-terminal region of NCALD and HPCA appear to be critical for interaction with S100B based on in vitro experiments. The possible physiological significance of this interaction is discussed.     

Study of the oriented overgrowth of sodium nitrate crystals

Formation of overgrowths of NaNO₃ on crystalline and noncrystalline substrates has been investigated employing the solvent evaporation method. Calcite, Gypsum, Sodium chloride, Mica, Glass and Polystyrene films formed on glass have been employed as substrate materials. The solvents employed are water, liquid ammonia, methyl alcohol and n-butyl alcohol. Random orientation of the overgrowths is observed on the substrates except in the case of calcite and mica. It is found that the morphology of the overgrowths manifests in the form of rhombic-, triangular- and hexagonal platelets. The occurrence of (001), (110) and (111) twins is observed. The effect of supersaturation and nature of the substrate on the formation of overgrowths is considered.     

Growth and characterization of nonlinear optical single crystals of l-arginine diiodate

Growth of good optical quality single crystals of l-arginine diiodate (abbreviated as LADI), a semiorganic nonlinear optical (NLO) material is reported. Crystals of dimension up to were obtained from its aqueous solution by slow solvent evaporation technique. The crystals were characterized by single crystal XRD, FTIR, optical absorption spectrum, microhardness, dielectric and photoconductivity studies. The DTA and TGA traces of LADI confirm the decomposition of the sample at 145 °C.     

Diffuse waves in a random medium layer with rough boundaries

The problem of scattering from a random medium layer with rough boundaries is formulated as an integral equation in which the random fluctuations are represented as a zero-mean random operator. The analysis for the diffuse fields is based on the ladder-approximated Bethe-Salpeter equation. An integral equation for the diffuse intensities thus derived displays the various multiple-scattering processes involved in our problem. Transport equations are also derived and several special cases are considered to illustrate the characteristics of the results and to compare them with those in the literature.     

Dielectric properties of Ba3Sr2DyTi3V7O30 ceramics

A polycrystalline sample, Ba₃Sr₂DyTi₃V₇O₃₀, with tungsten bronze structure was prepared by a mixed-oxide method at high temperature (950°C). Preliminary structural analysis of the compound showed an orthorhombic crystal structure at room temperature. Surface morphology of the compound was studied by scanning electron microscopy. The dielectric anomaly at 321°C may be attributed to the ferro-paraelectric phase transitions. This was also confirmed from the appearance of a hysteresis loop. The nature of variation of the ac conductivity and value of activation energy at different temperature regions, suggest that the conduction process is of mixed-type (i.e., ionic-polaronic and space charge generated from the oxygen ion vacancies).     

Comparison of denitrification between Paracoccus sp. and Diaphorobacter sp

Denitrification was compared between Paracoccus sp. and Diaphorobacter sp. in this study, both of which were isolated from activated sludge of a denitrifying reactor. Denitrification of both isolates showed contrasting patterns, where Diaphorobacter sp. showed accumulation of nitrite in the medium while Paracoccus sp. showed no accumulation. The nitrate reduction rate was 1.5 times more than the nitrite reduction in Diaphorobacter sp., as analyzed by the resting state denitrification kinetics. Increasing the nitrate concentration in the medium increased the nitrite accumulation in Diaphorobacter sp., but not in Paracoccus sp., indicating a branched electron transfer during denitrification. Diaphorobacter sp. was unable to denitrify efficiently at high nitrate concentrations from 1 M, but Paracoccus sp. could denitrify even up to 2 M nitrate. Paracoccus sp. was found to be an efficient denitrifier with insignificant amounts of nitrite accumulation, and it could also denitrify high amounts of nitrate up to 2 M. Efficient denitrification without accumulation of intermediates like nitrite is desirable in the removal of high nitrates from wastewaters. Paracoccus sp. is shown to suffice this demand and could be a potential organism to remove high nitrates effectively.