Evaluation of the authenticity of haloarchaeal strains by random-amplified polymorphic DNA

PCR analysis was performed on several archaeal species using 11 different 10-mer oligonucleotides of arbitrary sequence. Duplicates of the type strain of Halococcus morrhuae (ATCC 17082 and CCM 537) showed markedly different RAPD profiles. Moreover, RAPD patterns from the type strain of Haloferax denitrificans ATCC 35960 and the strain ATCC 49116 deposited as 'Haloferax larsenii' , were identical. This method represents an invaluable instrument in microbial screening and can clarify problems with strain identities.     

Genetic variation for foot-rot and Fusarium head-blight resistances among full-sib families of a self-incompatible winter rye (Secale cereale L.) population

The amount of genetic variation for resistance to foot rot caused by Pseudocercosporella herpotrichoides, Fusarium spp., and Microdochium nivale and for resistance to head blight caused by Fusarium culmorum are important parameters when estimating selection gain from recurrent selection in winter rye. One-hundred and eighty-six full-sib families of the selfincompatible population variety Halo, representing the Petkus gene pool, were tested for foot-rot resistance at five German location-year combinations (environments) and for head-blight resistance in three environments with artificial inoculation in all but one environment. Foot-rot rating was based on 25 stems per plot scored individually on a 1–9 scale. Head-blight resistance was plotwise scored on a 1–9 scale and, additionally, grain-weight per spike was measured relative to the non-inoculated control plots. Significant estimates of genotypic variance and medium-sized heritabilities (h ²=0.51–0.69) were observed in the combined analyses for all resistance traits. In four out of five environments, the amount of genetic variance was substantially smaller for foot-rot than for head-blight rating. Considerable environmental effects and significant genotype-environment interactions were found for both foot-rot and head-blight resistance. Coefficients of error-corrected correlation among environments were considerably closer than phenotypic correlations. No significant association was found between the resistances to both diseases (r=-0.20 to 0.17). In conclusion, intra-population improvement by recurrent selection should lead to substantial higher foot-rot and head-blight resistances due to significant quantitative genetic variation within Halo. Selection should be carried out in several environments. Lack of correlation between foot-rot and head-blight resistance requires separate infection tests for improving both resistances.     

A novel 145 kd brain cytosolic protein reconstitutes Ca(2+)-regulated secretion in permeable neuroendocrine cells.

The regulated secretory pathway is activated by elevated cytoplasmic Ca2+; however, the components mediating Ca2+ regulation have not been identified. In semi-intact neuroendocrine cells, Ca(2+)-activated secretion is ATP- and cytosol protein-dependent. We have identified a novel brain protein, p145, as a cytosolic factor that reconstitutes Ca(2+)-activated secretion in two neuroendocrine cell types. The protein is a dimer of 145 kd subunits, exhibits Ca(2+)-dependent interaction with a hydrophobic matrix, and binds phospholipid vesicles, suggesting a membrane-associated function. A p145-specific antibody inhibits the reconstitution of Ca(2+)-activated secretion by cytosol, indicating an essential role for p145. The restricted expression of p145 in tissues exhibiting a regulated secretory pathway suggests a key role for this protein in the transduction of Ca2+ signals into vectorial membrane fusion events.     

Supramaximal cycle tests do not detect seasonal progression in performance in groups of elite speed skaters.

Seven female and eight male elite junior skaters performed cycle ergometer tests at four different times during the 1987/1988 season. The tests consisted of a Wingate-type 30-s sprint test and a 2.5-min supramaximal test. The subjects were tested in February, May and September 1987 and in January 1988. Maximal oxygen consumption was measured during the 2.5-min test. With the exception of the maximal oxygen consumption of the women in May which was about 6% lower than in the other three tests, no seasonal changes in the test results could be observed--this, in spite of a distinct increase in training volume (from 10 to more than 20 h.week-1) and training intensity in the course of the season. When the test data were compared to those of elite senior skaters, it appeared that the junior skaters showed the same values for mean power output during the sprint test [14.2 (SD 0.4) W.kg-1 for the men and 12.6 (SD 0.5) W.kg-1 for the women] and maximal oxygen consumption [63.1 (SD 2.8) ml.kg-1.min-1 for the men and 55.3 (SD 3.5) ml.kg-1.min-1 for the women, respectively] as found for senior skaters. It seemed, therefore, that the effects of training in these skaters had already levelled off in the period before they participated in this investigation. In contrast to previous studies, no relationship could be shown between the test results and skating performance. This was most likely due to the homogenous character of the groups (mean standard deviations in power and oxygen consumption were only 5%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of hypoxia and fatty acids on the distribution of metabolites in rat heart

The effects of exogenous fatty acids and hypoxia on cardiac energy metabolism were studied by measuring mitochondrial and cytosolic adenine nucleotides as well as CoA and carnitine esters using a tissue fractionation technique in non-aqueous solvents. During normoxia, the administration of 0.5 mM palmitate caused a considerable increase in acyl-CoA and acylcarnitine, particularly in mitochondria. High-energy phosphates, however, were only slightly altered. A 90 min low-flow hypoxia caused a dramatic increase in mitochondrial acyl esters. The mitochondrial ATP content decreased significantly, while the cytosolic concentration was only slightly diminished, suggesting an inhibition of mitochondrial adenine nucleotide translocation by long-chain acyl-CoA. Addition of palmitate during hypoxia amplified hypoxic damage and reduced adenine nucleotides in both compartments considerably, while fatty acid metabolites were only slightly affected. In presence of an inhibitor of fatty acid oxidation (BM 42.304), the fatty-acid-induced acceleration of cardiac injury was prevented. Since BM 42.304 decreased mitochondrial acylcarnitine and increased the cytosolic concentration significantly, BM 42.304 was presumed to inhibit mitochondrial acylcarnitine translocase. However, a causal relationship between lipid metabolites and ischemic damage seemed unlikely.     

The role of active forms of oxygen in platelet aggregation and deaggregation

The effect of hydrogen peroxide on ADP-induced platelet aggregation in the presence of active oxygen species scavengers was studied. It was shown that the superoxide radical and singlet oxygen, alongside with hydrogen peroxide, may play a role in platelet interactions.     

Sequence-selective topoisomerase II inhibition by anthracycline derivatives in SV40 DNA: relationship with DNA binding affinity and cytotoxicity

Topoisomerase II mediated double-strand breaks produced by anthracycline analogues were studied in SV40 DNA. The compounds included doxorubicin, daunorubicin, two doxorubicin stereoisomers (4'-epimer and beta-anomer), and five chromophore-modified derivatives, with a wide range of cytotoxic activity and DNA binding affinity. Cleavage of 32P-end-labeled DNA fragments was visualized by autoradiography of agarose and polyacrylamide gels. Structure-activity relationships indicated that alterations in the chromophore structure greatly affected drug action on topoisomerase II. In particular, removal of substituents on position 4 of the D ring resulted in more active inducers of cleavage with lower DNA binding affinity. The stereochemistry between the sugar and the chromophore was also essential for activity. All the active anthracyclines induced a single region of prominent cleavage in the entire SV40 DNA, which resulted from a cluster of sites between nucleotides 4237 and 4294. DNA cleavage intensity patterns exhibited differences among analogues and were also dependent upon drug concentration. Intensity at a given site depended on both stimulatory and suppressive effects depending upon drug concentration and DNA sequence. A good correlation was found between cytotoxicity and intensity of topoisomerase II mediated DNA breakage.