Inhibitory action of selenite on Escherichia coli, Proteus vulgaris, and Salmonella thompson

Weiss, K. F. (Iowa State University, Ames), J. C. Ayres, and A. A. Kraft. Inhibitory action of selenite on Escherichia coli, Proteus vulgaris, and Salmonella thompson. J. Bacteriol. 90:857-862. 1965.-The resistance of three microorganisms, Escherichia coli (ISU-41), Proteus vulgaris (ISU-37c), and Salmonella thompson (ISU-86-2), to increasing concentrations of selenite was determined. E. coli was completely inhibited by 1.25% sodium hydrogen selenite, and 0.25% sodium hydrogen selenite caused a pronounced lag. P. vulgaris survived selenite concentrations of over 3%. S. thompson was inhibited completely by 3% selenite but not by 2.5%, although there was a considerable lag and a decrease in total growth. The relationship of growth, uptake, and reduction of selenite was determined. The susceptible E. coli incorporated up to twice as much selenium as did the other two organisms during the early stages of incubation. Radioautographs of seleno analogues of sulfur-containing amino acids revealed the presence of seleno-cystine in all three organisms, and seleno-methionine in E. coli. Compounds having R(F) values corresponding to possible oxidation products of seleno-methionine were present in the hydrolysates of P. vulgaris and S. thompson. Kinetic aspects of selenite uptake, rather than the ultimate localization of selenite in the cell protein, appear to be the factors that determine the degree of resistance or of susceptibility to selenite.     

STUDIES ON CELL DEFORMABILITY : I. Effect of Surface Charge

The deformability of the surface membranes of Sarcoma 37 and Ehrlich murine ascites tumor cells was assessed by the pressure required to suck a hemispherical bulge from these cells into a micropipette. It was shown that treatment with neuraminidase allowed the cells to be deformed with significantly less suction, and that enzymatic treatment also produced a significant reduction in surface charge as determined by measurement of cellular electrophoretic mobility. It is suggested that the increase in cellular deformability may be related to charge reduction, and that the charge at the cell periphery may affect not only the magnitude of the potential energy barriers hindering contact between cells, but also the ease with which cells can form low radius of curvature probes in order to help overcome these barriers.     

Structural Changes in Mammalian Cells Associated with Cooling to -79°C

Suspensions of HeLa and S37 cells, with and without added glycerol, were cooled in stages to -79°C. and held at that temperature for 30 minutes. After warming to room temperature the cells were fixed, sectioned, and compared by phase contrast and electron microscopy with similar specimens kept at room temperature. Correlated viability tests were made. Abnormal cytological characteristics, visible with the phase contrast microscope, were clearly related to the sequence of freezing and thawing, and the proportion of altered cells was highest in specimens cooled without glycerol. Electron microscopy showed that even in the presence of glycerol all cells were markedly altered, with distinctive vesiculation and disruption of the various intracellular membranes. There is evidence that much cytoplasmic damage is compatible with survival, but it seems likely that separation of the two layers of the nuclear envelope and rearrangement of the nuclear contents are signs of irreversible damage. The findings lend some support to the belief that cell death on cooling is due largely to denaturation of semipermeable membranes, caused by the increasing concentration of electrolytes.     

Polarisationsoptischer Nachweis von Neurosekret mit N,N′-Diäthylpseudoisozyaninchlorid

Zusammenfassung An neurosekretorischen Zellen des Nucleus praeopticus der Bachforelle wurde nach oxydativer Vorbehandlung die Anlagerung von N,N-Diäthylpseudoisozyaninchlorid vergleichend fluoreszenz-und polarisationsmikroskopisch untersucht. Dicht angeordnete dissoziierte Sulfonsäuregruppen binden die Farbstoffassoziate elektrostatisch. Eine orientierte Einlagerung der reversiblen Polymerisate des Pseudoisozyanins in die neurosekretorischen Zellen ist polarisationsoptisch nachweisbar. Die polarisationsmikroskopischen Erscheinungen entsprechen den fluoreszenzoptischen. Gleichartige Anwendung von Akridinderivaten bei niedrigem pH-Wert und histochemische Kontrollversuche unterstützen die angegebene Erklärung der Befunde. Auf die Möglichkeit der polarisationsoptisch-quantitativen Untersuchung des Neurosekrets wird hingewiesen.

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Summary A comparative study (fluorescence and polarization microscopy) was carried out in the neurosecretory cells of the nucleus praeopticus in trouts. The object of the investigation was to study the adsorption of N,N-diethyl pseudoisocyanine chloride following oxidation. Densely arranged dissociated sulphonic acid groups seem to bind the dye electro-staticly. An orientated inclusion of the reversible polymers of the pseudoisocyanine into the neurosesecretory cells is demonstrable by polarization microscopy. The polarization optical image corresponds to the fluorescence microscopic image. The results of a similar investigation using acridine derivatives at a low pH and histochemical controls seem to support the results of the findings. The possibility of quantitative polarization optical studies of the neurosecretion is discussed.

     

Biomechanical interactions of cancer cells with the microvasculature during metastasis

Metastasis is a major, life-threatening complication of cancer. The bloodstream is the most important disseminative route for cancer cells liberated from their parent tumors. Single circulating cancer cells are arrested in the microvasculature, where the vast majority are killed by rapid or slow processes, and the relatively few survivors grow into micrometastases. We review the underlying causes of one type of rapid cancer cell death in the microcirculation, namely, that caused by biomechanical interactions of cancer cells with microvessel walls, which may result in cell surface membrane expansion and lethal rupture. These lethal interactions appear to be important rate-regulators in hematogenous metastasis, and to dictate some aspects of metastatic patterns. Although these are not the only interactions involving cancer cells, in contrast to others involving cellular and humoral defense mechanisms, they have received comparatively little attention.     

A physical map including a new class I gene (cda12) of the human major histocompatibility complex (A2/B13 haplotype) derived from a monosomy 6 mutant cell line

To avoid interpretative problems due to restriction fragment length polymorphisms, the monosomy 6 mutant cell line BM19.7 was employed to establish a molecular map of the human major histocompatibility (HLA) complex in the A2,B13,Bw4,DRw6,DRw52,DQw1,DPw2 haplotype. Results were obtained mainly by field-inversion gel electrophoresis and Southern blotting techniques. The map extends to 4800 kb and includes the HLA complex with a length of 4200 kb. Five HTF islands could be positioned on the map. The class I region has a size of about 2000 kb and includes nonclassical HLA class I genes, some of which must be localized within 200 kb telomeric of HLA-A. A new class I gene, cda12, distinct from HLA-A, HLA-B, or HLA-C, has been localized within 50 kb from HLA-A. The class I region contains a gap of about 500 kb, just telomeric of HLA-C, in which further class I genes could not be detected. The class II region has a size of 1000 kb, which is separated from the class I region by about 1200 kb. The 5' end of the HLA-B gene is situated centromeric, giving an orientation opposite to that of the TNFA and TNFB loci. The estimated length of the HLA complex correlates well with its size determined cytogenetically using mutant cell lines with interstitial deletions.