Significance of the inactivation of transport in thermal death of Escherichia coli.

Cells of Escherichia coli ML308-225, harvested from the exponential phase, were heated in 50 mM potassium phosphate, and the loss in viability and inability to transport lactose, proline, and alpha-methylglucoside was compared. After cells were heated at 48 degrees C for 15 min, there was a 16% loss in viability and a similarly small reduction in the steady-state accumulation of lactose at 25 degrees C. The initial rates of lactose and proline transport were severely inhibited by heating at either 48 or 50 degrees C, but substantial recovery occurred within 5 to 7 min at 25 degrees C. Heating at 50 degrees C for 15 min caused an 86% loss in viability, but only a 53% decrease in the steady-state accumulation of lactose and only a 24% reduction in the initial rate of alpha-methylglucoside uptake. Twice as much alpha-methylglucoside was accumulated at 50 degrees C as at 25 degrees C. Although alpha-methylglucoside phosphate leaked from the cells at 50 degrees C, the concentration retained within the cells was about 500 times that externally, when only about 14% of the cells were viable. Overall, these results indicate that cells made nonviable by heating at 50 degrees C still have significant membrane integrity.     

Quantification of two viruses in technical preparations of Orgyia pseudotsugata baculovirus by means of buoyant density centrifugation of viral deoxyribonucleic acid.

A reliable method was developed for the quantitative determination of two nuclear polyhcdrosis viruses present in commercially prepared viral insecticides used against Orgyia pseudotsugata. Deoxyribonucleic acids, from nuclear polyhedrosis bundle virus and nuclear polyhedrosis single-rod virus, were separated on CsCl gradients according to their respective buoyant densities, 1.715 and 1.704 g/ml. The proportions of the two viruses were quantified by measuring the relative absorbance at 254 nm of their deoxyribonucleic acid peaks.     

Cytochemische Untersuchungen über den Zusammenhang von nativer Gärungsintensität und desoxygenem Energiepotential beim anoxybiotischen Phosphatstoffwechsel

Zusammenfassung In vorliegenden Untersuchungen konnte an drei gärungsphysiologisch distinkten Zellkategorien der Nachweis geliefert werden, daß-nach 5-bzw. 24-stündiger anaerober phosphatischer Inkubierung, ausgehend vom P-Mangelstadium — in der ersten Gruppe (stark bis mittelstark gärender Hefetyp) eine M-T-Färbung von stark bis mittelstark eintritt, die in der zweiten Gruppe (mäßig bis schwach gärender Hefetyp) auf mäßig bis schwach absinkt, während in de dritten Gruppe (nicht gärender Hefetyp) der M-T-Test vollkommen negativ ausfällt. Die markant sich abzeichnenden cytologischen Befunde stehen in guter Übereinstimmung mit den gleichzeitig durchgeführten chemischen Kontrollanalysen.Als gesetzmäßiges Reaktionsprinzip wurde erkannt, daß die Intensität der anaeroben P-Regenerierung nach P-Mangelzüchtung abhängig ist vom Grad des nativen Spaltungsvermögens der Gärungszellen, welches seinerseits einen Maßstab für die Aktivierbarkeit des anaeroben Energiepotentials bildet. Dabei kommt jedoch der Gärung, was durch anabiotische Versuche sichergestellt wurde, keine ergonische Funktion in bezug auf die phosphorylierende P-Transferierung unter anoxybiotischen Bedingungen zu.     

The quantification of formazans in tissue sections by microdensitometry. I. The use of neotetrazolium chloride

Synopsis This article describes the use of a microdensitometer for the measurement of formazan deposits in tissue sections. Some examples are given to illustrate the various applications of this technique in the assessment of glucose-6-phosphate dehydrogenase activity. These are (1) the separate measurement of the red half-formazan intermediate and purple diformazan of neotetrazolium, and the effect of incubation time on their production, (2) the measurement of activities in different regions of the liver lobule, and the selective effect of phenobarbitone, and (3) the measurement of enzyme activity in individual cartilage cells in normal and osteoarthrosis-prone animals. All activities can be expressed in absolute units as nmol hydrogen/mm³/hr, and thus compared with standard biochemical data. The activities obtained all fall within the range of published values for biochemical systems.Paper given at the Royal Microscopical Society's European Histochemistry Meeting at Nottingham in September 1975.     

Characterization of ribonucleolytic activity of angiogenin towards tRNA

Yeast tRNA is a convenient substrate for the assay of the ribonucleolytic activity of human angiogenin. The optimal pH, [NaCl], and temperature for tRNA cleavage by angiogenin are approximately 6.8, 15-30 mM, and approximately 55 degrees C, respectively, as compared with approximately 8.0, 100-200 mM, and approximately 65 degrees C, respectively, for RNase A. Polyanions and metals both inhibit angiogenin and RNase A but to different extents.