Search PubMed⌕ Search

Biomedical subjects

R Scherrer

Publications and source records attributed to R Scherrer.

53 records · Page 3Linked to original sources

Location of calcium within Bacillus spores by electron probe x-ray microanalysis.

Spectroscopic microanalysis of the element-characteristic X rays produced by a scanning electron microprobe was employed to detect calcium and carbon in both intact and thin-sectioned spores of Bacillus cereus T and B. megaterium QM B1551. Linear scan profiles and multilinear scan images of the X-ray emissions for calcium (Ca(Kalpha)) were compared with those for carbon (C(Kalpha)) as an index of mass. Location was accomplished by stereological comparisons with secondary electron images and conventional transmission electron micrographs. Although the elements could be detected at the attogram level theoretically, spatial resolution was limited to approximately 500 to 1,000 nm in an intact spore, e.g., by the primary electron beam diameter, the electron-excited spore microvolume, and the type of specimen support. The resolution was improved to approximately 100 to 200 nm by use of thin-sectioned spores, with precautions to prevent calcium leakage from the specimen during preparations. In both intact and sectioned spores, calcium was distributed throughout the spore, similarly to carbon, and concentrated mainly in a central region corresponding to the spore protoplast.

Bacillus cereus↗

Molecular sieving by the Bacillus megaterium cell wall and protoplast.

Passive permeabilities of the cell wall and protoplast of Bacillus megaterium strain KM were characterized by use of 50 hydrophilic probing molecules (tritiated water, sugars, dextrans, glycols, and polyglycols) which varied widely in size. Weight per cent uptake values (R(w)) were measured at diffusional equilibrium under conditions that negated the influences of adsorption or active transport. Plots of R(w) for intact cells as a function of number-average molecular weight ( M(n)) or Einstein-Stokes hydrodynamic radius ( r(ES)) of the solutes showed three phases: a protoplast uptake phase with a polydisperse exclusion threshold of M(n) = 0.6 x 10(3) to 1.1 x 10(3), r(ES) = 0.6 to 1.1 nm; a cell wall uptake phase with a polydisperse exclusion threshold of M(n) = 0.7 x 10(5) to 1.2 x 10(5), r(ES) congruent with 8.3 nm; and a total exclusion phase. Isolated cell walls showed only the latter two phases. However, it became evident that the cell wall selectively passed only the smallest molecules in a heterodisperse polymer sample. When the molecular-weight distributions of polyglycol samples ( M(n) = 1,000, 1,450, and 3,350) were determined by analytical gel chromatography before and after uptake by intact cells or isolated cell walls, a quasi-monodisperse exclusion threshold was obtained corresponding to M(n) = 1,200, r(ES) = 1.1 nm. The permeability of isolated protoplasts was assessed by the relative ability of solutes to effect osmotic stabilization. An indefinite exclusion threshold, evident even with monodisperse sugars, was attributed to lengthwise orientation of the penetrating rod-shaped molecules. Altogether, the best estimate of the limiting equivalent porosity of the protoplast was 0.4 to 0.6 nm in radius and of the cell wall, 1.1 nm.

Bacillus megaterium↗

Macromolecular sieving by the dormant spore of Bacillus cereus.

The threshold surface porosity in the dormant spore of Bacillus cereus strain T was assessed by measuring passive permeabilities to a series of polydisperse polyethylene glycol samples which increased in average molecular size. The apparent exclusion threshold at diffusional equilibrium corresponded to a polymer of number-average molecular weight ( M(n)) = 150,000 and equivalent hydrodynamic radius ( r(ES)) = 16 nm, which confirmed a previous report. However, analytical gel chromatography before and after uptake by the spores revealed that only the low molecular weight fractions in a polymer sample distribution were taken up. From graphical analyses of the changes in molecular weight distributions, a quasi-monodisperse exclusion threshold was determined corresponding to M(n) = 8,000 and r(ES) = 3.2 nm. Thus, the equivalent porosity in the limiting outer integument appeared much more restrictive than heretofore shown for spores, although still more open than the monodisperse equivalent for the cell wall of vegetative bacilli.

Bacillus cereus↗

Differentiation of mouse plasmocytomas in vitro: two phenotypically stabilized variants of the same cell.

Mouse myelomas from Balb/c and C3H mice were established in tissue culture for more than 200 passages. When allowed to attach to a surface, they differentiated into two stabilized forms, one of which was fibroblast-shaped, grew without contact inhibition, and could be transplanted back to mice. The other had the morphology of epithelial cells, showed contact inhibition, and was not transplantable back to mice. For one strain (MOPC 173) it was demonstrated that both types of cells synthesize molecules with idiotypic determinants of the original myeloma protein. The relationship between the host cells and a leukemia-type virus present in the original tumor cells has been studied during different stages of cellular differentiation.

Animals↗

Viral and host deoxyribonucleic acid synthesis in Shope fibroma virus-infected cells as studied by means of high-resolution autoradiography.

Incorporation of (3)H-thymidine by BSC-1 cells infected with Shope fibroma virus was studied by means of high-resolution electron microscopic radioautography. One-hour pulses with the radioactive precursor were given at various times after infection, during a one-step growth cycle of the virus. In the cytoplasm of infected cells, reacted grains occurred over foci of viroplasm; these foci are believed to represent the true sites of viral deoxyribonucleic acid (DNA) replication. Shope fibroma virus DNA synthesis began before 3 hr postinfection, reached a maximum at 8 to 9 hr, and then declined rapidly. It was demonstrated that the decline in (3)H-thymidine uptake is correlated with the onset of viral morphogenesis. In comparison with the noninfected culture, the nuclear labeling, which reflects host DNA metabolism, was slightly reduced by 4 hr postinfection. Inhibition became more marked as infection progressed, and host DNA synthesis was almost completely suppressed in late stages of viral development.

Animals↗