Partial cloning and expression of the cyclin B gene in the ovary of the bullseye puffer (Sphoeroides annulatus)

The bullseye puffer is a marine fish species with great potential for aquaculture in Mexico, and the understanding of its reproductive physiology at every level of biological organization is essential in order to succeed. Several molecules orchestrate the complex process of oocyte maturation and spawning. One of these molecules is cyclin B, which is the regulatory subunit of the maturation-promoting factor. In this study, a fragment of the cyclin B gene was isolated from the ovary of the bullseye puffer using an RT-PCR approach. The gene fragment was homologous to the cyclin B2 gene of other vertebrate species. Similar levels of cyclin B gene expression were detected in ovaries at different developmental stages, except for atretic ovaries from captive fish which did not spawn. However, cyclin B gene expression was maintained in captive fish treated with LHRH-a to induce spawning, and appeared to be similar to the pattern observed in wild fish. It is possible that the reduced expression of cyclin B in atretic ovaries is the result of mRNA degradation during atresia. Alternatively, reduced gene expression could be a controlling factor in the process of oocyte reabsorption since cyclin B is required for final oocyte maturation and ovulation.     

Biochemical characterisation and kinetic properties of a purified lipase from Aspergillus niger in bulk phase and monomolecular films

An isolate of Aspergillus niger was used as source of lipase which was purified to a specific activity of 729 U/mg. It has an acidic pH optimum and has a half-life of 42 h at pH 4.4, which can be increased to 138 h in the presence of 10 mM calcium ions. For the first time a lipase from Aspergillus niger was characterised using the monomolecular film technique. The lipase was classified to have a sn-1 selectivity using diacylglycerols and R-isomer hydrolytic preference with pseudolipids representing triacylglycerols in which two of the ester bonds were replaced with ether and amide linkages.     

Field microplot performance of the peach-almond hybrid GF-677 after inoculation with arbuscular mycorrhizal fungi in a replant soil infested with root-knot nematodes

The effects of arbuscular mycorrhizae (AM) on the development and nutrition of the peach almond hybrid GF-677 rootstock in a replant soil heavily infested with Meloidogyne javanica were evaluated in field microplot conditions for two growing seasons. There was a significant beneficial effect of mycorrhizal inoculation on plant growth and nutrition in previously pasteurized replant soil. In natural replant soil, early inoculation with a mixed AM inoculum of Glomus intraradices, Glomus mosseae and Glomus etunicatum did not affect growth parameters. Whilst inoculation with these AM fungi led to suppression of root-knot nematode reproduction, natural mycorrhizal colonization of the replant soil with native AM fungi did not. Accepted: 6 December 2000     

A membrane-associated protein with Cr(VI)-reducing activity from Thermus scotoductus SA-01

A membrane-associated chromate reductase from Thermus scotoductus SA-01 has been purified to apparent homogeneity and shown to couple the reduction of Cr(VI) to NAD(P)H oxidation, with a preference towards NADH. The chromate reductase is a homodimer with a monomeric molecular weight of 48 kDa and a noncovalently bound FAD coenzyme. The enzyme is optimally active at a pH of 6.5 and 65 degrees C with a K(m) of 55.5+/-4.2 microM and a V(max) of 2.3+/-0.1 micromol Cr(VI) min(-1) mg(-1) protein. The catalytic efficiency (k(cat)/K(m)) of the enzyme was found to be comparable to that found for quinone reductases but more efficient than the nitroreductases. N-terminal sequencing and subsequent screening of a genomic library of T. scotoductus revealed an ORF of 1386 bp, homologous (84%) to the dihydrolipoamide dehydrogenase gene of Thermus thermophilus HB8. These results extend the knowledge of chromate reductases mediating Cr(VI) reduction via noncovalently bound or free redox-active flavin groups and the activity of dihydrolipoamide dehydrogenases towards physiologically unrelated substrates.     

Changes in mitochondrial function induced by dequalinium precede oxidative stress and apoptosis in the human prostate-cancer cell line PC-3

Mitochondria play central roles in diverse physiological and pathological conditions associated with cell survival and death. Delocalized lipophilic cations, such as dequalinium (DQA), are accumulated in cancer cells attracted by the highly negative mitochondrial transmembrane potential of these cells. DQA showed a potent anticancer activity in cells from different malignancies. Here, we report the effect of DQA on PC-3 prostate cancer cells. Incubation with DQA at concentrations between 1.5 and 100 μM from 24 to 48 h decreases cell viability. The decrease in cell viability together with a loss of mitochondrial transmembrane potential induced an increase in reactive oxygen species production and cell death via caspase-3 dependent apoptotic pathway. DQA was shown to cause moderate to strong cell death in a time and concentration dependent manner, causing a most advantageous effect at a concentration of 10 μM applied for a long 48 h time period, which might be a consequence of the kinetics of intracellular DQA accumulation in mitochondria, but also of the mechanisms of DQA-induced cell death. This data shows DQA as a promising agent against the human prostate cancer PC-3 cell line, activating the caspase-3 dependent apoptotic pathway. This fact might be beneficial for possible future applications in cancer therapy.     

Die Culturversuche Heinricher's mitAlectorolophus und deren Bedeutung für die Systematik der Gattung

Ohne Zusammenfassung     

Heart Rate and Systolic Blood Pressure Variability in the Time Domain in Patients with Recent and Long-Standing Diabetes Mellitus

Diabetes Mellitus (DM) affects the cardiovascular response of patients. To study this effect, interbeat intervals (IBI) and beat-to-beat systolic blood pressure (SBP) variability of patients during supine, standing and controlled breathing tests were analyzed in the time domain. Simultaneous noninvasive measurements of IBI and SBP for 30 recently diagnosed and 15 long-standing DM patients were compared with the results for 30 rigorously screened healthy subjects (control). A statistically significant distinction between control and diabetic subjects was provided by the standard deviation and the higher moments of the distributions (skewness, and kurtosis) with respect to the median. To compare IBI and SBP for different populations, we define a parameter, α, that combines the variability of the heart rate and the blood pressure, as the ratio of the radius of the moments for IBI and the same radius for SBP. As diabetes evolves, α decreases, standard deviation of the IBI detrended signal diminishes (heart rate signal becomes more “rigid”), skewness with respect to the median approaches zero (signal fluctuations gain symmetry), and kurtosis increases (fluctuations concentrate around the median). Diabetes produces not only a rigid heart rate, but also increases symmetry and has leptokurtic distributions. SBP time series exhibit the most variable behavior for recently diagnosed DM with platykurtic distributions. Under controlled breathing, SBP has symmetric distributions for DM patients, while control subjects have non-zero skewness. This may be due to a progressive decrease of parasympathetic and sympathetic activity to the heart and blood vessels as diabetes evolves.