Biomedical subjects

M M Krishna

Publications and source records attributed to M M Krishna.

6 recordsLinked to original sources

PubMed2001

Rotational dynamics of surface probes in lipid vesicles.

Translational and rotational diffusion of fluorescent molecules on the surface of small biological systems such as vesicles, proteins and micelles depolarize the fluorescence. A recent study has treated the case of the translational dynamics of surface probes (M.M.G. Krishna, R. Das, N. Periasamy and R. Nityananda, J. Chem. Phys., 112 (2000) 8502-8514) using Monte Carlo and theoretical methods. Here we extend the application of the methodologies to apply the case of rotational dynamics of surface probes. The corresponding fluorescence anisotropy decays were obtained using the Monte Carlo simulation methods for the two cases: surface probes undergoing rotational dynamics on a plane and on a sphere. The results were consistent with the theoretical equations which show that Monte Carlo methods can be used to simulate the surface diffusion problems. The anisotropy decay for the rotational diffusion of a molecule on a planar surface is single exponential and the residual anisotropy is zero. However, residual anisotropy is finite for the case of rotational diffusion on a sphere because of the spatial averaging of the anisotropy function. The rotational correlation time in both the cases is (4Drot)(-1) with Drot being the rotational diffusion coefficient. Rotational dynamics of a surface bound dye in a single giant liposome and in sonicated vesicles were studied and the results were explained according to the theoretical equations. A fast component of fluorescence depolarization was also observed for sonicated vesicles which was interpreted as wobbling-in-cylinder dynamics of the surface-bound dye.

Carbocyanines↗

PubMed2001

Hydrogen exchange.

Explore the source record for details and available documents.

Biochemistry↗

PubMed1999

Location and orientation of DODCI in lipid bilayer membranes: effects of lipid chain length and unsaturation.

The location and orientation of a linear dye molecule, DODCI, in lipid bilayer membrane were determined by the effect of viscosity and refractive index of the aqueous medium on the fluorescence properties of the dye bound to the membrane. The membrane-bound dye is solubilized in two sites, one near the surface (short fluorescence lifetime) and another in the interior of the membrane (long lifetime). The ratio of the dye in the two locations and the orientation of the dye (parallel or perpendicular to the membrane) are sensitive to the lipid chain length and unsaturation in the alkyl chain. The fraction of the dye in the interior region is higher for short alkyl chains (C12>C14>C16>>C18C20) and in unsaturated lipids (C14:1>C14:0, C16:1>C16:0). These experimental results are consistent with the general principle that the penetration of an amphiphilic organic molecule in the interior region of the membrane is more when the structure of th bilayer is more fluid-like.

Carbocyanines↗

PubMed1997

Spectrally constrained global analysis of fluorescence decays in biomembrane systems.

Dynamic and steady-state fluorescence spectroscopic properties of a dye probe measured in a multicomponent biological system are often required to be separated into the spectra and lifetimes of individual spectroscopically distinct species. The conventional method of obtaining decay-associated spectra fails when the lifetimes of the fluorophore in the membrane phase and in the aqueous phase are very close to each other. This paper describes a global analysis method which takes advantage of the known spectrum and lifetime of the dye in the aqueous phase. This method is used to identify the spectra for two fluorescent species (lifetimes, 0.84 and 1.97 ns) of the dye DODCI in EggPC vesicle membranes by keeping the spectrum and lifetime (0.68 ns) of the dye in the aqueous phase as fixed parameters. The structural identity of the two membrane-bound dye species was established by the effect of refractive index and/or viscosity of the aqueous medium on the lifetimes.

Carbocyanines↗

PubMed1996

Cell surface properties of Clostridium difficile: haemagglutination, relative hydrophobicity and charge.

Five well characterised strains of Clostridium difficile of differing virulence and two Escherichia coli strains, a verotoxigenic O157:H7 isolate and a urinary isolate, were examined for cell surface hydrophobicity and charge, and haemagglutinating ability. Phase partition in hexadecane or octan-1-ol was similar for C. difficile and E. coli, as was retention by hydrophobic interaction chromatography (HIC), indicating moderate hydrophobicity. The salt agglutination test showed E. coli to be hydrophobic and C. difficile to be hydrophilic. Relative hydrophobicity determined by HIC when charge effects were not nullified, i.e., to reflect more closely conditions in vivo, showed C. difficile to bind less well. Growth of C. difficile in caecal emulsions to simulate conditions in vivo did not alter the cell surface hydrophobicity. The phase partition method for charge determination indicated that E. coli and C. difficile had a net negative charge, although this was weaker for C difficile than E. coli. However, although E. coli exhibited a net negative charge as determined by immuno-gold electronmicroscopy (IGEM), in keeping with the results of the phase partition method, C. difficile was shown to be predominantly positively charged by IGEM, and by movement in a charged field as determined by paper electrophoresis and a novel method based on light microscope observation. A cell-wall deficient mutant of C. difficile was weakly positively charged, showing that most of the charge resides in the cell wall.

Cell Membrane↗

PubMed1992

Molecular, immunological, and biological characterization of a toxin A-negative, toxin B-positive strain of Clostridium difficile.

A cytotoxigenic Clostridium difficile strain that fails to produce toxin A but causes hemorrhage and bloody fluid accumulation in ligated ileal loops of rabbits and hemorrhage and diarrhea in hamsters is described. The lack of reaction of DNA from this strain in hybridization studies with a toxin A gene-specific 4.5-kb probe and polymerase chain reaction studies with six toxin A-specific primers indicate the absence of the toxin A gene. The cytotoxin produced by this strain was not responsible for the enterotoxic or hemorrhagic activity and shared characteristics with toxin B, i.e., its cytotoxicity was neutralized by antibodies to toxigenic strains of C. difficile and Clostridium sordellii. Polymerase chain reaction studies with toxin B-specific primers showed that the DNA from this strain produced a 690-bp product in addition to the expected 591-bp product.

Animals↗