Operational Sex Ratio and Individual Mating Frequencies in Two Bushcricket Species (Orthoptera,Tettigonioidea, Poecilimon)

During mating the male bushericket transfers a large spermatophore which is consumed by the female after copulation. Such a ‘nuptial gift' is likely to have strong effects on the mating frequency of both males and females and thus on the operational sex ratio (OSR): For the male a longer time period will be required in which he is prevented from further matings. For the female consumption may be advantageous increasing interest in matings. To quantify these effects field studies were carried out on two species of the genus Poecilimon. Both species differed widely in the mean number of matings observed per day (19% of all females in P. veluchianus compaied to 42% in P. affinis; Fig. 2) and thus in the distribution of female mating intervals (P. veluchianus 2 days, P. affinis 1 day (modes); Fig. 3). Separate experiments on the minimum male mating interval and the spermatophore weight also revealed distinct differences: the mean spermatophore weight was 26% of the male body weight in P. veluchianus and 15% in P. affinis (Fig. 6). Accordingly the refractory period was longer in P. veluchianus; 3 days after a previous mating nearh all males were ready to remate, compared to 2 days in P. affinis (Fig. 5). These differences, however, compensated one another in that the amount of spermatophylax material transferred per day was very similar in both species in relation to female body weight. Using data on female mating frequency per day and male remating ability, an upper estimate for the OSR was calculated. A lower limit for the OSR was given taking into account the most frequent female mating interval (big. 7). In both species the OSR remained relatively constant for large parts of the season. The medians (upper/lower estimate for the ratio males to females ready to mate: 2.95/1.19 in P. veluchianus; 2.22 0.88 in P. affinis) were very low, comparable to that in explosive breeding frog species. Thus in the bushcricket mating system, females may be able to demand large male mating efforts so that the operational sex ratio can come close to unity.     

Influence of nicotianamine and iron supply on formation and elongation of adventitious roots in hypocotyl cuttings of the tomato mutant 'chloronerva' (Lycopersicon esculentum)

The influence of nicotianamine (NA) on formation and elongation of adventitious roots in hypocotyls of de-rooted NA-less mutant seedlings of Lycopersicon esculentum Mill, was examined in relation to the iron supply [ferric N-N'-ethylenediaminedi-(2-hydroxyphenylacetate) (FEDDHA), ferric ethylenediaminetetracetate (FeEDTA), ferric N-(2-hydroxyethyl)-ethylenediaminetriacetate (FeHEDTA, Fe-citrate and FeCl₃] in the nutrient solution. The initiation of root primordia in hypocotyl cuttings was independent of NA and occurred with about the same frequency in both, mutant and wild-type. In the mutant the development of primordia to adventitious roots was blocked at all iron sources used, except FeEDTA. Addition of NA (5x 10⁻⁶ to 2 × 10⁻⁵ M ) to the rooting medium resulted in a fast growth of adventitious roots in mutant cuttings with all iron sources tested. Rooting of wild-type cuttings was independent from NA application and iron sources. We suppose that NA is involved in the intracellular transport of iron. Its function is possibly linked with chelation of ferrous iron in the cell.     

Cloning and characterization of a new polyketide synthase gene cluster in Streptomyces aureofaciens CCM 3239.

We cloned a new polyketide gene cluster, aur2, in Streptomyces aureofaciens CCM3239. Sequence analysis of the 9531-bp DNA fragment revealed 10 open reading frames, majority of which showed high similarity to the previously characterized type II polyketide synthase (PKS) genes. An unusual feature of the aur2 cluster is a disconnected organization of minimal PKS genes; ACP is located apart from the genes for ketosynthases KSalpha and KSbeta. The aur2 gene cluster was disrupted in S. aureofaciens CCM3239 by a homologous recombination, replacing the four genes (aur2A, E, F, G) including ketosynthase KSalpha, with antibiotic resistance marker gene. The disruption did not affect growth and differentiation, and disrupted strain produced spores with wild-type grey-pink pigmentation. The biochromatographic analysis of the culture extracts from S. aureofaciens wild type and aur2-disrupted strains did not reveal any difference in the pattern of antibacterial compounds.     

Immunodominant CD4+ T-cell epitope within nonstructural protein 3 in acute hepatitis C virus infection.

In acute hepatitis C virus infection, 50 to 70% of patients develop chronic disease. Considering the low rate of spontaneous viral clearance during chronic hepatitis C infection, the first few months of interaction between the patient's immune system and the viral population seem to be crucial in determining the outcome of infection. We previously reported the association between a strong and sustained CD4+ T-cell response to nonstructural protein 3 (NS3) of the hepatitis C virus and a self-limited course of acute hepatitis C infection. In this study, we identify an immunodominant CD4+ T-cell epitope (amino acids 1248 to 1261) that was recognized by the majority (14 of 23) of NS3-specific CD4+ T-cell clones from four of five patients with acute hepatitis C infection. This epitope can be presented to CD4+ T cells by HLA-DR4, -DR11, -DR12, -DR13, and -DR16. HLA-binding studies revealed a high binding affinity for 10 of 13 common HLA-DR alleles. Two additional CD4+ T-cell epitopes, amino acids 1388 to 1407 and amino acids 1450 to 1469, showed a very narrow pattern of binding to individual HLA-DR alleles. Our data suggest that the NS3-specific CD4+ T-cell response in acute hepatitis C infection is dominated by a single, promiscuous peptide epitope which could become a promising candidate for the development of a CD4+ T-cell vaccine.     

Biophoton emission

The phenomenon of ultraweak photon emission from living systems was further investigated in order to elucidate the physical properties of this radiation and its possible source. We obtained evidence that the light has a high degree of coherence because of (1) its photon count statistics, (2) its spectral distribution, (3) its decay behavior after exposure to light illumination, and (4) its transparency through optically thick materials. Moroever, DNA is apparently at least an important source, since conformational changes induced with ethidium bromide in vivo are clearly reflected by changes of the photon emission of cells. The physical properties of the radiation are described, taking DNA as an exciplex laser system, where a stable state can be reached far from thermal equilibrium at threshold.     

Influence of fatty acids on energy metabolism. 2. Kinetics of changes in metabolic rates and changes in subcellular adenine nucleotide contents and pH gradients following addition of octanoate and oleate in perfused rat liver

The uncoupling-like effect of fatty acids [ Scholz , R., Schwabe , U., and Soboll , S. (1984) Eur. J. Biochem. 141, 223-230] was further substantiated in experiments with perfused rat livers by two ways: firstly the kinetics of changes in metabolic rates (oxygen consumption, ketogenesis, fatty acid oxidation) were analysed; secondly subcellular contents of adenine nucleotides and pH gradients across the mitochondrial membrane were determined following fractionation of freeze-fixed and dried tissues in non-aqueous solvents. The following results were obtained. The relaxation kinetics of the increase in oxygen consumption following fatty acid infusion revealed two components, a rapid one with a half-time around 10 s and a slow one with a half-time of more than 100 s. The rapid component was similar to the kinetics of fatty acid oxidation (ketogenesis and 14CO2 production from labelled fatty acids) whereas the half-time of the slow component was in the range of half-times observed with the increase in oxygen consumption following addition of carbonylcyanide p-trifluoromethoxyphenylhydrazone. In the presence of fatty acids, the cytosolic ATP concentrations and ATP/ADP ratios decreased, whereas the corresponding parameters for the mitochondrial space were either increased (oleate) or decreased (octanoate). The effects of oleate were dependent on the albumin concentrations in the perfusate. The normally large difference between cytosolic and mitochondrial ATP/ADP ratios became smaller. Similar observations were obtained with uncoupling agents. The pH gradient across the mitochondrial membrane as calculated from the subcellular distribution of 5,5 dimethyl[2-14C]oxazolidine-2,4-dione was inversed following the addition of both carbonylcyanide p-trifluoromethoxyphenylhydrazone and fatty acids, i.e. the mitochondrial matrix became more acidic than the cytosol. The pH gradient was not affected when oleate was added in the presence of high albumin concentrations. The data support the hypothesis that the increase in hepatic oxygen consumption due to octanoate or oleate is, in part, caused by a mechanism similar to uncoupling of oxidative phosphorylation. This mechanism seems not to be an artifact of isolated systems; it may be of physiological importance for processes in which reducing equivalents are removed independently of the ATP demand of the hepatocyte.     

Ultrastructural immunocytochemical evidence for peptidergic neurotransmission in the pond snail Lymnaea stagnalis

Summary Three neuronal systems of the pond snail Lymnaea stagnalis were immunocytochemically investigated at the ultrastructural level with the unlabeled peroxidase-antiperoxidase technique. Preliminary electrophysiological and cell-filling investigations have shown that a cluster of neurons which reacts positively with an antiserum against the molluscan cardio-active peptide FMRFamide, sends axons to the penis retractor muscle. In this muscle anti-FMRF-amide (aFM) positive axons form neuro-muscular synapses with (smooth) muscle fibers. The morphological observations suggest the aFM immunoreactive system to be involved in peptidergic neurotransmission. In the right parietal ganglion a large neuron (LYAC) is penetrated by aFM positive axons which form synapse-like structures (SLS) with the LYAC. The assumption that the SLS represent the morphological basis for peptidergic transmission is sustained by the observation that iontophoretical application of synthetic FMRFamide depolarizes the LYAC. The axons of a group of pedal anti-vasopressin (aVP) positive cells run in close vicinity to the cerebral ovulation (neuro-)-hormone producing cell system (CDC system) Synapses or SLS between the two systems were not observed. The fact that (bath) application of arg-vasopressin induces bursting in the CDC, may indicate that the vasopressin-like substance of the aVP cells is released non-synaptically.     

Heterogeneity ofHLA defined in PLT: A cellular assay detects differences not seen using HLA-DRw serology

The primed lymphocyte typing test (PLT) is used to detect the gene products of theHLA-D region which are responsible for secondary restimulation of cells primed in MLC. Alternatively, products of theHLA-D region may be detected serologically using antisera directed against a subpopulation of lymphocytes; these are the so-called DRw determinants. The PLT was used to see if it were possible to detect heterogeneity within a given serologically defined group using a cellular test. As priming combinations, we used family members identical for one haplotype and differing in theHLA-A, B andC regions, but not theD region of the second haplotype. Our results indicated that it was possible to prime against this second haplotype and that the segregation of the difference followedHLA. Therefore, using a cellular test it was possible to detect differences among cells belonging to a given DRw group. This suggests that PLT can be a useful tool to identify those serological groups which are composed of heterogenous determinants. In addition, it points out the problem in using any one test to establish identity of theHLA-D region, especially for clinical purposes.