Virulence studies on chromosomal α-toxin and Θ-toxin mutants constructed by allelic exchange provide genetic evidence for the essential role of α-toxin in Clostridium perfringens-mediated gas gangrene

The pathogenesis of clostridial myonecrosis, or gas gangrene, involves the growth of the anaerobic bacterium Clostridium perfringens in the infected tissues and the elaboration of numerous extracellular toxins and enzymes. The precise role of each of these toxins in tissue invasion and necrosis has not been determined. To enable genetic approaches to be used to study C. perfringens pathogenesis we developed an allelic exchange method which involved the transformation of C. perfringens cells with a suicide plasmid carrying a gene insertionally inactivated with an erythromycin-resistance determinant. The frequency with which double reciprocal crossover events were observed was increased to a workable level by increasing the amount of homologous DNA located on either side of the inactivated gene. Allelic exchange was used to isolate mutations in the‘chromosomal pfoA gene, which encodes an oxygen-labile haemolysin known as Θ-toxin or perfringolysin O. and in the chromosomal pic gene, which encodes the α-toxin or phospholipase C. The resultant mutants failed to produce detectable Θ-toxin or α-toxin activity, respectively, and could be complemented by recombinant plasmids that carried the respective wild-type genes. The resultant strains were virulence tested in a mouse myonecrosis model. The results showed that the pic mutants had demonstrably reduced virulence and therefore provided definitive genetic evidence for the essential role of α-toxin in gas gangrene or clostridial myonecrosis.     

Biomechanical and Morphometric Differences in Triticum aestivum Seedlings Differing in Rht Gene-Dosage

Biomechanical and morphometric comparisons among coleoptilesfrom wheat seedlings differing in Rht gene-dosage (Rht = 0,2, 4 doses) are presented in an effort to evaluate the influenceof Rht on the mechanics of soil penetration by this organ. Rhtis known to reduce seedling establishment compared to the wildtype. Data from 3–7-day-old seedlings indicate that Rhtreduces tissue elastic modulus E, increases the second momentof area I, and decreases the slenderness ratio (l/r) of coleoptiles.Rht-relatedchanges in E and I are such that the flexural stiffness of coleoptilesfrom Rht plants does not differ significantly from the wildtype-hence the growing coleoptiles of all three genotypes haveequivalent biomechanical capacity to penetrate the soil. Rhtreduction of coleoptile slenderness ratios confers a capacityto safely sustain higher axial compressive loads compared tocoleoptiles with equivalent flexural stiffness but higher ratios.However, wild type seedlings produce longer coleoptiles andlonger subcrown internodes than Rht seedlings. Longer coleoptilesdeliver the crown node closer to the top of the soil beforethe crown node extends beyond the lateral confinement of thecoleoptile. This reduces the potential for buckling of the subcrowninternode and leaves due to the compressive loading of soil.Rht affects a variety of mechanical features whose influenceis dependent upon the stage of seedling growth and the degreeof soil compaction. However, at equivalent depths of burialwhich exceed the maximum length of coleoptiles and moderatesoil compaction, Rht is biomechanically disadvantageous to seedlingestablishment. Wheat, germination, biomechanics, Rht-gene     

Neuroendocrine mediated effects of electromagnetic-field exposure: possible role of the pineal gland

Reports from recent epidemiological studies have suggested a possible association between extremely low frequency (ELF; including 50- or 60-Hz) electric- and magnetic-field exposure, and increased risk of certain cancers, depression, and miscarriage. ELF field-induced pineal gland dysfunction is a possible etiological factor in these effects. Work in our laboratory and elsewhere has shown that ELF electromagnetic-field exposure can alter the normal circadian rhythm of melatonin synthesis and release in the pineal gland. Consequences of reduced or inappropriately timed melatonin release on the endocrine, neuronal, and immune systems are discussed. Laboratory data linking ELF field exposure to changes in pineal circadian rhythms in both animals and humans are reviewed. The authors suggest that the pineal gland, in addition to being a convenient locus for measuring dyschronogenic effects of ELF field exposure, may play a central role in biological response to these fields via alterations in the melatonin signal.     

Destabilization of secondary structure in polyadenylic acid by formaldehyde

Results of circular dichroism and ultraviolet absorption measurements of poly A in neutral aqueous formaldehyde solution are presented, which show that the hydroxymethylated residues retain the ability to stack, but the stacking tendency is diminished. The midpoint of the thermal denaturation profile falls from about 38°C to about 18°C on complete hydroxymethylation. Hydroxymethylation, however, appears to have only a small effect on the apparent standard enthalpy and entropy of stacking, both of which become slightly more negative. When present at concentrations greater than about 2 g/100 ml, formaldehyde has a further and somewhat more dramatic denaturing effect on poly A. This ability is probably related to the chemical similarity of aqueous formaldehyde (methylene glycol) and ethylene glycol, which is known to be an effective denaturant for nucleic acids. In the presence of 20 g/100 ml formaldehyde, a very broad thermally induced transition of poly A is observed through the loss of ultraviolet hypochromism. Only the low-temperature part of the transition is observed; it appears to be completed only at temperatures well over 100°C where the hypochromism with respect to adenosine has largely disappeared. The transition cannot be identified with the unstacking reaction usually observed through thermal denaturation experiments in the absence of formaldehyde, although the associated structure may be related in some way. The molecular nature of the structure associated with the transition is not known.     

Vacuum ultraviolet circular dichroism of poly(galacturonic acid), sodium polygalacturonate and calcium polygalacturonate

Vacuum ultraviolet circular dichroism spectra are reported for poly(galacturonic acid) solution and film, sodium polygalacturonate solution and film, and calcium polygalacturonate gel. In addition to the positive c.d. band near 208 nm previously observed, we find a pair of higher energy bands at 170 180 nm (negative) and 145 nm (positive). The low energy band, assigned to an $\text{n-π}{}^1$ carboxyl transition, is blue-shifted upon gelation or film formation.