Dna stability and survival of bacillus subtilis spores in extreme dryness

The inactivation of Bacillus subtilis spores during long-term exposure (up to several months) to extreme dryness (especially vacuum) is strain-dependent, through only to a small degree. During a first phase (lasting about four days) monolayers of spores lose about 20% of their viability, regardless of the strain studied. During this phase loss in viability can be equally attributed both to damages of hydrophobic structures (membranes and proteins) and DNA. During a second phase lasting for the remaining time of experimental observation (weeks, months and years) the loss in viability is slowed. A viability of 55% to 75% (depending on the strain) is attained after a total exposure of 36 days. The loss in viability during the second phase can be correlated with the occurrence of DNA double strand breaks. Also covalent DNA-protein cross-links are formed by vacuum exposure. If the protein moiety of these cross-links is degraded by proteinase K-treatment in vitro additional DNA double strand breaks result. The data are also discussed with respect to survival on Mars and in near Earth orbits.     

Identification of mitochondrial proteins on two-dimensional electrophoresis gels of extracts of adult Drosophila melanogaster

Several hundred proteins have been resolved on two-dimensional gels of extracts of [³⁵S]methionine-labeled adult Drosophila melanogaster. 27 of these polypeptides disappear from the gel pattern after feeding the K⁺ ionophore nonactin. These proteins have been identified as mitochondrial, since the two-dimensional gel pattern of extracts of isolated mitochondria correlates well with the pattern of the proteins missing from that of nonactin-treated flies. Nine new proteins also appear on the two-dimensional gels of the extracts from the nonactin-treated flies. Apparently, these nine proteins are precursors of the mature mitochondrial forms. These particular data support the concept that processing of many of the cytoplasmically synthesized mitochondrial proteins requires a specific membrane potential, and that some of these proteins are modified intramitochondrially. However, using [³⁵S]methionine incorporation techniques, not all labeled polypeptides disappear from mitochondria during such treatment. Feeding similarly radiolabeled flies with chloramphenicol, an inhibitor of mitochondrial protein synthesis, results in the disappearance of only one protein from the gel pattern with the concurrent appearance of a ‘new’ high-molecular-weight polypeptide. Collectively, these data show that a specific group of [³⁵S]methionine-labeled mitochondrial proteins can be identified by selective inhibition of mitochondrial function in whole cell protein maps of adult D. melanogaster.     

In-situ investigations on small-scale local and short-term changes of sublittoral macrobenthos in Lübeck Bay (western Baltic Sea)

In-situ studies on sublittoral soft bottom macrofauna (depth: 14–16 m) employing the underwater laboratory (UWL) “Helgoland” were carried out. Sets of samples were compared for small-scale local and short-term changes in species richness, faunal abundance, numerical dominance, diversity, evenness, homogeneity, and similarity. It could be shown that minor differences in sediment quality can cause conspicuous heterogeneity within a small sampling area (diameter: ca. 100 m). Both spatfall and mortality of benthic invertebrates can change the faunal structure within a short period (two months). The degree of change varies between species and thus at stations harbouring different faunal assemblages as well.     

The impact of ticks on pheasant territoriality

Pheasants are competent reservoir hosts for the Lyme disease spirochaete, Borrelia burgdorferi s.l., and carry large, but highly over-dispersed, infestations of the vector ticks, Ixodes ricinus . The effects of experimental reduction of tick infestation levels on the survival and territorial behaviour of male pheasants were studied. Over three years in two woodlands in southern England, birds were marked individually and half were fitted with a slow-release acaricide, which substantially reduced their tick burdens from March to August. Acaricide treatment affected reproductive success but had no discernible impact on the survival rates of male pheasants. The degree of wattle inflation by males, an indicator of territorial status and a correlate of harem acquisition, was significantly greater among treated males. In each year, a significantly higher proportion of treated (overall 44%) than control (22%) males acquired harems. Males that acquired females ranged over small areas on field edges. By contrast, those with no females ranged more widely in woods and the adjoining fields, increasing their exposure to questing ticks. The relative contribution of such roving males to tick-borne pathogen transmission may thus increase.     

Effects of nucleotides and divalent cations on phospholipase activity in Saccharomyces cerevisiae

Divalent cations activate the lysophospholipase and transacylase reactions catalyzed by the same enzymes in the yeast Saccharomyces cerevisiae. The activation was observed at neutral pH, but not at the pH optimum of lysophospholipase/transacylase, near 3.5. Adenine nucleotides, especially AMP and ADP, are strong inhibitors of the same group of enzymes. Half maximal inhibition by AMP was found at a concentration of about 20 M. The inhibition by nucleotides in low concentrations is enhanced by divalent cations.     

Aestival dormancy in the cabbage moth Mamestra brassicae L. (Lepidoptera: Noctuidae)

Summary In a geographically wide distribution the life cycles of different populations of the cabbage moth Mamestra brossicae are adapted to a remarkable diversity of climatic conditions. This is undoubtedly a proof of its success in adaptation. Some populations living in regions characterized by a drought period interrupting the growth season are capable of distinguishing between one critical day length signalling the onset of the drought period and another signalling the end of the growth season. This study, therefore, is primarily concerned with the geographical patterns in the variability of the adaptional responses of populations exposed to environmental conditions requiring different strategies and tactics in, synchronizing individual, life cycles. It is also a contribution to our understanding of evolutionary mechanisms maintaining median responses to photoperiodically inductive day lengths in geographically different populations. The populations investigated originated from regions differing in predictability of the incidence, onset and duration of a drought period: Freiburg (48.0°N, Southern Germany), Avignon (44.0°N, Southern France), and Argelès (42.5°N, Southern France). Geographical variation with respect to both onset and duration of a drought period consequently results in clinal variation of the variability of innate day length thresholds triggering aestival dormancy and of innate duration of aestivation. In this paper we considered the influence of geographically changing temperatures on aestival dormancy induction. Even in southern populations of M. brassicae a temperature dependent switch off-mechanism exists which prevents aestival dormancy under certain environmental conditions. The effective temperatures vary geographically, too. What the geographical patterns in adaptive responses really are, is discussed.This research was supported by the Deutsche Forschungsgemeinschaft (Sa 259/3-1)     

Stachyose synthesis in mature leaves of Cucumis melo. Purification and characterization of stachyose synthase (EC 2.4.1.67)

Galactinol: raffinose-6-galactosyltransferase (EC 2.4.1.67), a stachyose synthase, was extracted from mature leaves of Cucumis melo cv. Ranjadew and was purified to homogeneity by (NH₄)₂SO₄ precipitation, ion-exchange chromatography, gel-filtration and non-denaturing polyacrylamide gel electrophoresis. A specific activity of 516 kat · mg^-1 and a 160-fold purification was achieved. The pH optimum of the enzyme reaction was found to be 6.8 in sodium-phosphate buffer, and the temperature optimum 32° C. The purified enzyme was very sensitive towards SH-poisons but its reaction was hardly affected by changes in the ion composition of the assay medium. The two-substrate enzyme was specific for galactinol and raffmose; uridine-diphosphate galactose and p-nitrophenyl--d-galactoside as well as melibiose were not accepted by the purified enzyme. Stachyose synthesis was competitively inhibited by concentrations >4 mM raffinose as well as 2.5 mM galactinol. The K _m values determined under non-saturating conditions were 3.3 mM for raffinose and 7.7 mM for galactinol. Myoinositol was a strong competitive inhibitor with a K _i of 1.8mM. Galactinol was hydrolyzed in the absence of raffinose with a K _m of 0.8 mM. The pure enzyme is a protein with a molecular weight of at least 95 kDa and an isoelectric point of 5.1. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the presence of two subunits of 45 and 50 kDa. Polyclonal antibodies from rabbit were obtained which were specific for the native enzyme but cross-reacted with other proteins separated under denaturing conditions.Abbreviations DEAE diethylaminoethyl - DTT dithiothreitol - FPLC fast protein liquid chromatography - HPLC high-performance liquid chromatography - PAGE polyacrylamide gel electrophoresis - SDS sodium dodecyl sulfate This work was supported by Deutsche Forschungsgemeinschaft. The gift of galactinol by Dr. T. Schweizer (Nestlé, Switzerland) is gratefully acknowledged.     

Biosynthesis of gallotannins

Cell-free extracts from leaves of Rhus typhina L. (sumach) were found to transfer the 1-O-galloyl moiety of l,6-di-O-galloyl-β-d-glucose to the 2-position of the same compound, yielding 1,2,6-tri-O-galloyl-β-d-glucose and leaving 6-O-galloylglucose as the deacylated by-product. The enzyme catalyzing this ‘disproportionation’ was purified almost 1700-fold. It had a molecular weight of approx. 56 000, a K _m value of 11.5 mM, was stable between pH 4.5 and 6.5, and most active at pH 5.9 and 40° C. The systematic name “1,6-di-O-galloyl-glucose: 1,6-di-O-galloylglucose 2-O-galloyltransferase” (EC 2.3.1.) was proposed for this new enzyme whose detection provided evidence that, in addition to β-glucogallin (1-O-galloyl-β-d-glucose), higher substituted glucose esters also have the potential to serve as acyl donors in the biosynthesis of gallotannins.     

No evidence for linkage of autosomal dominant proximal spinal muscular atrophies to chromosome 5q markers

Summary Two recent articles have reported the linkage of a gene for recessive spinal muscular atrophy (SMA) on the chromosome region 5q11.2–13.3. Our data show no linkage of the dominantly inherited forms of SMA to this chromosome region.     

B lymphocyte receptors and polyphosphoinositide degradation

Resting B lymphocytes can be activated and induced to proliferate by antibodies against their antigen receptors (anti-lg). We demonstrate an early increase in the level of [3H]inositol trisphosphate in [3H]inositol-labeled murine B cells, which suggests breakdown of phosphatidylinositol bisphosphate by phospholipase C. In line with this, the level of [3H]1,2-diacylglycerol was also elevated after incubation of [3H]arachidonic-acid-labeled B cells with anti-Ig. Anti-lg also caused a rapid increase in the level of cytosolic Ca2+ in B cells. In contrast, two other polyclonal B cell activators, lipopolysaccharide and phorbol myristate acetate, failed to induce any of these effects. Our results suggest that anti-lg may induce B cell growth via phosphoinositide degradation and Ca2+ mobilization, and that phorbol myristate acetate, and possibly lipopolysaccharide, bypass these initial events.