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Biomedical subjects

K B Easterbrook

Publications and source records attributed to K B Easterbrook.

At least 19 recordsLinked to original sources

Two bacteriophages of Clostridium difficile.

Two temperate bacteriophages of differing morphology and host range were isolated by screening 94 isolates of Clostridium difficile. Phage 41 had a 300-nm flexible tail, whereas phage 56 had a shorter tail with a contractile sheath. Electron microscopy of phage 56 lysates exposed to elevated magnesium concentrations showed small virus-like particles which were 21 nm in diameter. The addition of MgCl2 to semisolid agar overlays enhanced both the titer and plaque size of phage 56. Phage 56 was more temperature labile than phage 41 and demonstrated unusual lability in buffer at pH 7.0. One-step growth and adsorption experiments revealed that both phages had latent periods of about 60 min, but phage 56 adsorbed to its indicator strain more efficiently. Phage 56, which was obtained from a toxigenic strain of C. difficile, was used to lysogenize its nontoxigenic indicator strain, but no conversion to toxigenicity was observed in this strain.

Bacteriophages↗

Physicochemical and immunological homogeneity of spinin, the subunit-protein of bacterial spinae.

Bacterial spinae from marine bacterium D71 are multi-subunit structures of a single protein. This protein, called spinin, is homogeneous by immunodiffusion and immunoelectrophoresis, amino acid composition, polyacrylamide gel electrophoresis with a number of buffer systems, sedimentation velocity and diffusion boundary analysis. Sedimentation equilibrium gives Mr = 19,000, while phosphate polyacryl-amide gel electrophoresis in presence of dodecyl sulfate gives Mr = 32,000. The lower Mr estimate for spinin is supported by sedimentation equilibrium in 6 M guanidine . HCl, and covalent cross-linking with dimethyl suberimidate or glutaraldehyde. The higher Mr value probably arises from an anomalous spinin-dodecyl sulfate interaction. Isoelectric focusing in polyacrylamide gel gives pI = 3.45; however, the focusing pattern also contains three distinct bands that may arise from hydrolysis of the spinin protomer during anodic migration. This study presents the first extensive physicochemical characterization of spinin and provides the basis for investigating the subunit assembly of spinae.

Amino Acids↗

Susceptibility of human embryonic kidneys in organ culture to herpesvirus hominis.

101 pairs of human embryonic kidneys of 5-12 weeks' gestation were maintained in whole organ culture by our previously described technique, with herpesvirus hominis types1 and 2 added to our regular media. One kidney of each pair served as a control and was exposed to identical culture medium without virus. Cultures were maintained for 24-120 h to study the time sequence of viral infectivity. Organs were then examined for the presence of virus by electron microscopy and histochemical staining. Histological studies of virus-infected kidneys showed either (1) complete organ death or (2) virus localization in undifferentiated cells, plus disorganization of architecture in differentiated areas. Control organs showed normal organization.

Culture Media↗

The attachment of bacterial spinae.

Spinae are attached to protease-sensitive structural proteins in the external surface of the outer membrane. Agents and (or) treatments affecting ionic, hydrophobic, or hydorgen bonds are ineffective in releasing spinae from bacteria. As judged by thin-sectioning and freeze-fracturing techniques, the outer membrane is not modified at the attachment site to a detectable extent, and the other surface layers are not involved. The attachment of spinae is thus differentiated from that of flagella.

Binding Sites↗

Arrangement of morphological subunits in bacterial spinae.

The filament, that is helically arranged to form the bacterial spina, is composed of morphological subunits (oligomers) about 5.6 nm in width and 11 nm in length. The oligomers are asymmetrical in that the inner surface is grooved. Image analysis of negative-stained spinae ribbons indicates that the oligomers are paired, possibly beaded structures, the arrangement of which is easily distorted during preparation. In intact spinae, the oligomer orientation may be normal to the filament axis, but in collapsed freeze-etched spinae, the oligomers are inclined at a constant angle of about 72 degrees to the filament axis.

Bacterial Proteins↗

Application of freeze-etching method to the study of reovirus-infected LLC-MK2 cells.

A continuous cell line of rhesus monkey kidney cells, LLC-MK(2), was infected with reovirus type 1 (Lang). The cells were freeze-etched as a method for observing the structural details of the reovirus-induced cytoplasmic inclusion. Information on maturation may be obtained by preparing infected cells for electron microscopy by freeze-etching.

Animals↗