Heat-induced temperature sensitivity of outgrowing Bacillus cereus spores

Inactivation of Bacillus cereus spores during cooling (10 degrees C/h) from 90 degrees C occurred in two phases. One phase occurred during cooling from 90 to 80 degrees C; the second occurred during cooling from 46 to 38 degrees C. In contrast, no inactivation occurred when spores were cooled from a maximum temperature of 80 degrees C. Inactivation of spores at a constant temperature of 45 degrees C was induced by initial heat treatments from 80 to 90 degrees C. The higher temperatures accelerated the rate of inactivation. Germination of spores was required for 45 degrees C inactivation to occur; however, faster germination was not the cause of accelerated inactivation of spores receiving higher initial heat treatments. Repair of possible injury was not observed in Trypticase soy broth (BBL Microbiology Systems), peptone, beef extract, starch, or L-alanine at 30 or 35 degrees C. Microscopic evaluation of spores outgrowing at 45 degrees C revealed that when inactivation occurred, outgrowth halted at the swelling stage. Inhibition of protein synthesis by chloramphenicol at the optimum temperature also stopped outgrowth at swelling; thus protein synthesis may play a role in the 45 degree C inactivation mechanism.     

Small reactive carbonyl compounds as tissue lipid oxidation products; and the mechanisms of their formation thereby

Small reactive carbonyl compounds (RCCs) such as formaldehyde, acetaldehyde, acrolein, crotonaldehyde, glyoxal, methylglyoxal, glycolaldehyde, glycidaldehyde, and 2-butene-1,4-dial are involved in carbonyl and oxidative stress-related physiological disorders. While some evidence indicates that lipid oxidation could be an important source of these compounds in vivo, this has sometimes been doubted because the mechanisms of their formation thereby are poorly understood. Here, representative literature supporting the significant formation of these compounds during lipid oxidation under physiologically relevant conditions are highlighted, and the strengths and weaknesses of previously proposed mechanisms of their formation thereby are considered. In addition, based on the current understanding of lipid oxidation chemistry, some new pathways of their formation are suggested. The suggested pathways also generate 4-hydroxy-2-butenal, a precursor of the carcinogen furan, whose endogenous formation in tissues has hitherto not been seriously considered.     

NMR spectroscopy of 113Cd(II)-substituted gene 32 protein

Gene 32 protein (g32P), the single-stranded DNA binding protein from bacteriophage T4, contains 1 mol of Zn(II)/mol of protein. This intrinsic zinc is retained within the DNA-binding core fragment, g32P-(A+B) (residues 22-253), obtained by limited proteolysis of the intact protein. Ultraviolet circular dichroism provides evidence that Zn(II) binding causes significant changes in the conformation of the peptide chain coupled with alterations in the microenvironments of tryptophan and tyrosine side chains. NMR spectroscopy of the 113Cd(II) derivative of g32P-(A+B) at both 44.4 and 110.9 MHz shows a single 113Cd resonance, delta 637, a chemical shift consistent with coordination to three of the four sulfhydryl groups in the protein. In vitro mutagenesis of Cys166 to Ser166 creates a mutant g32P that still contains 1 Zn(II)/molecule. This mutant protein when substituted with 113Cd(II) shows a 113Cd signal with a delta and a line width the same as those observed for the wild-type protein. Thus, the S-ligands to the metal ion appear to be contributed by Cys77, Cys87, and Cys90. Relaxation data suggest that chemical shift anisotropy is the dominant, but not exclusive, mechanism of relaxation of the 113Cd nucleus in g32P, since a dipolar modulation from ligand protons is observed at 44.4 MHz but not at 110.9 MHz. Complexation of core 113Cd g32P with d(pA)6 or Co(II) g32P with poly(dT) shows only minor perturbation of the NMR signal or d-d electronic transitions, respectively, suggesting that the metal ion in g32P does not add a ligand from the bound DNA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of the transciption inhibitory protein, TF1, on phage SP01 promoter complex formation and stability.

The uptake of a homologous single-stranded fragment by superhelical DNA produces a complex that contains a stable displacement loop. When the circular DNA was relaxed by the random action of pancreatic DNAase, complexes dissociated by a process which requires that the single-stranded arm of the D-loop be intact. We attribute the dissociation to branch migration, the exchange of like strands at a branch point. The kinetics of dissociation were biphasic. A fraction of the nicked complexes dissociated in a few seconds, the rest dissociated much more slowly. The fraction of molecules that dissociated slowly was directly related to the length of the third strand, and inversely related to temperature. Salt also inhibited dissociation. Under physiological conditions, 37 °C and 0.15 m-NaCl, more than half of complexes containing a third strand of 1000-nucleotide residues survived for at least one minute. These observations provide a guide to handling certain natural or synthetic branched derivatives of DNA. Analyzing our data by the method of Thompson et al. (1976), we have estimated that the time for the exchange of one nucleotide for another at a single-stranded branch is 12 microseconds; but the calculated value depends strongly upon the assumption that single-strand branch migration occurs by a random walk.     

Molecular basis of biodegradation of chloroaromatic compounds

Chlorinated aromatic hydrocarbons are widely used in industry and agriculture, and comprise the bulk of environmental pollutants. Although simple aromatic compounds are biodegradable by a variety of degradative pathways, their halogenated counterparts are more resistant to bacterial attack and often necessitate evolution of novel pathways. An understanding of such evolutionary processes is essential for developing genetically improved strains capable of mineralizing highly chlorinated compounds. This article provides an overview of the genetic aspects of dissimilation of chloroaromatic compounds and discusses the potential of gene manipulation to promote enhanced evolution of the degradative pathways.     

Factors influencing the fecundity of Moniliformis moniliformis (Acanthocephala): constant diet and varied dose

Aspects of the reproductive performance of Moniliformis moniliformis were investigated in rats allowed to feed ad libitum on a purified diet containing 1% (w/w) fructose as an energy source for the worms. The rats were infected with either 10, 20, 40 or 80 cystacanths each with the intention of investigating density-dependent effects on worm fecundity. The establishment of the worms in the gut was independent of dose, but survival, growth and reproductive performance generally were shown to be related to the infective dose given to the rats. The effects could not be related to the absolute numbers of worms present in the small intestine at post-mortem examination. In general, some unidentified regulatory process appeared to operate to create severe density-dependence in survival so that surviving parasites were not present in numbers expected to generate competition. Attainment of sexual maturity, growth and the production of mature eggs by worms from rats given doses of 80 cystacanths each were delayed compared with worms from rats given the other doses, but eventually the performance of the high-dose worms caught up. Worms attached more anteriorly in the small intestine grew bigger and produced more mature eggs. Possible mechanisms responsible for the observed effects are discussed.