A physical genome map of Pseudomonas aeruginosa PAO.

A complete macrorestriction map of the 5.9 Mb genome of Pseudomonas aeruginosa PAO (DSM 1707) was constructed by the combination of various one- and two-dimensional pulsed field gel electrophoresis techniques. A total of 51 restriction sites (36 SpeI sites, 15 DpnI sites) were placed on the physical map yielding an average resolution of 110 kb. Several genes encoding virulence factors and enzymes of metabolic pathways were located on the anonymous map by Southern hybridization. Distances between the gene loci were similar on the genetic and physical maps, suggesting an even distribution of genome mobility throughout the bacterial chromosome. The four rRNA operons were organized in pairs of inverted repeats. The two-dimensional macro-restriction techniques described herein are generally applicable for the genome mapping of any prokaryote and lower eukaryote which yields resolvable fragment patterns on two-dimensional pulsed field gels.     

The heat generated by yeast cultures with a mixed metabolism in the transition between respiration and fermentation

The heat generated by both batch and continuous cultures of the yeast K. fragilis was studied using a modified Bench Scale Calorimeter. Batch cultures were used to measure the heat dissipation rates and the heat yields during fully aerobic and completely anaerobic growth, whereas continuous cultures enabled, in addition, a quantitative study of heat dissipation rates during growth on mixed metabolism. In this case, the extent of fermentation versus respiration could be specified and controlled by varying the degree of oxygen limitation. The heat dissipated per unit biomass formed was highest for fully respirative catabolism and fell continuously to a much lower value typical of anaerobic cultures as the catabolism was shifted increasingly to the fermentative mode. The heat generated per mole of oxygen taken up stayed quite close to the fully aerobic value of 506 kJ mol(-1) even when a sizable fraction of the substrate available to catabolism was fermented. If the fraction of respiration in the metabolism is lowered beyond a certain threshold, the ratio of the heat generation to oxygen consumption starts to increase dramatically and finally tends to infinity for fully anaerobic growth. All experimental results were quantitatively analyzed and explained on the basis of a simple model which formally describes the cultures in terms of two parallel "chemical" reactions. In simple cases such as the one presented here, the model enables calculation of the whole stoichiometry of the culture from a single measured heat yield.     

Perturbation in liver mitochondrial Ca2+ homeostasis in experimental iron overload: a possible factor in cell injury

The functional state of isolated mitochondria and specifically the integrity of the inner membrane, were investigated in the liver of rats made siderotic by dietary supplementation with carbonyl iron. The concentration of iron in the hepatic tissue increased progressively up to nearly 40 days and reached a steady-state level. When the iron content reached a threshold value (higher than 90 nmol/mg protein) the occurrence of in vivo lipid peroxidation in the mitochondrial membrane was detected. This process did not result in gross alterations in the mitochondrial membrane, as indicated by electron microscopy, phosphorylative capability and membrane potential measurements. On the contrary, the induction of lipoperoxidative reaction appeared to be associated with the activation of Ca2+ release from mitochondria. This was shown to occur as a consequence of rather subtle modifications in the inner membrane structure via a specific efflux route, which appeared to be linked to the oxidation level of mitochondrial pyridine nucleotides. The induction of this Ca2+ release from iron-treated mitochondria resulted in enhancement of Ca2+ cycling, a process which dissipates energy to reaccumulate into mitochondria the released Ca2+. The perturbation in mitochondrial Ca2+ homeostasis reported here may be a factor in the onset of cell damage in this experimental model of hepatic iron overload.     

Microdissection of the Prader-Willi syndrome chromosome region and identification of potential gene sequences

The Prader-Willi syndrome chromosome region on the long arm of human chromosome 15 was microdissected and microcloned from 20 GTG-banded metaphase chromosomes, and 5000 recombinant clones were obtained. Of these clones, 39% identify single-copy human DNA sequences, most of which map to the dissected chromosome region and are evolutionarily conserved in other species. Three of eleven clones studied in detail are deleted in several patients with Prader-Willi syndrome. The microclones will be useful for the physical characterization of the Prader-Willi syndrome chromosome region and the identification of the affected genes in this disease.     

Comparison of 16S rRNA sequences from the family Pasteurellaceae: phylogenetic relatedness by cluster analysis

The taxonomy of the family Pasteurellaceae has remained controversial despite investigations of biochemistry, serology, and nucleic acid relatedness. In an attempt to resolve some of this confusion, we have partially sequenced the 16S rRNAs of seven members of the family, representing all three genera. The sequences were aligned, similarity scores calculated, and single, average and complete linkage cluster analysis of the resulting distance matrix performed. In this way, an evolutionary branching pattern of these closely related species was reconstructed, and the approximate phylogenetic position of the family determined. Actinobacillus (Haemophilus) actinomycetemcomitans clustered with Haemophilus instead of Actinobacillus, supporting transfer of this species to the genus Haemophilus. Thus cluster analysis of phylogenetic relatedness was found to be particularly useful for studying closely related organisms, and could be performed using a microcomputer.     

A new gene for resistance to green leafhopperNephotettix virescens (Distant) in rice

The mode of inheritance of resistance to green leafhopper in 12 cultivars of riceOryza saliva L. was studied. Seedlings of parent and hybrid populations were artificially infested with second- and third-instar virus-free green leafhopper nymphs. Seedling reaction was scored when TNI, the susceptible check, was completely killed. The results revealed that single dominant genes confer resistance in six varieties, two independent dominant genes in four varieties, and single recessive genes in two varieties. The single dominant genes in Sri Gaya, ARC 7320, and T23 and one of the two genes in Aswina and Bhura Rata 2 are allelic toGlh-1; while Bhawalia hasGlh-5 gene. The second gene of Bhura Rata 2 is allelic to IR28 gene. Resistance in Chamar is controlled by two independent genes one of which is allelic toGlh-5 and the other allelic to IR28 gene. Bazal hasGlh-2 andGlh-5. The single recessive gene in ARC 7012 is allelic toglh-4 but the single recessive gene in DV85 is nonallelic to and independent ofglh-4. This new recessive gene is designated asglh-8. The single dominant genes of Dumai, Gadur, and the second gene of Aswina are nonallelic to all the known genes for resistance.     

On the occurrence and biology of some Antarctic Mysidacea (Crustacea)

Summary 10 species of Mysidacea were sampled during the Polarstern and Walther Herwig SIBEX cruises to the Antarctic Peninsula region from 1983 to 1986. The commonest were Antarctomysis maxima, A. ohlini and Mysidetes posthon. For these species the size and maturity stage composition as well as the length-weight relationships are given. The species considered are growing slowly and attain a long life span, M. posthon reached age class 3+, A. ohlini at least lived as long as 5 years, while A. maxima was found to be of age 5+ in the Peninsula area and probably 6+ in the Weddell Sea. The generation time of Antarctomysis species was 4 years and 3 years in Mysidetes posthon. Fecundity was low, mean number of offspring was about 13 for M. posthon and 21 for A. maxima. Rematuration was observed for the species A. maxima, indicating several spawning events during its life span. Biomass production is low for A. maxima, shown by a P/B-index of 0.98. Mortality of this species was estimated to be Z=1.1, which indicates that 33% of the specimens of an age group survive until the next year. The distribution and spatial separation of the two Antarctomysis species is discussed.