Immunocytochemical colocalization of adhesive proteins with clathrin in human blood platelets: further evidence for coated vesicle-mediated transport of von Willebrand factor,fibrinogen and fibronectin

Coated membranes and vesicles play an important role in receptor-mediated endocytosis and intracellular trafficking in various cell types, and are also present in blood platelets. Platelets take up certain proteins from the blood plasma, such as von Willebrand factor and fibrinogen, and these substances are transferred to storage granules. The receptors for these plasma proteins on the platelet plasma membrane have been well characterized, but morphological evidence for their transport to the storage granules is not yet available. In an attempt to clarify this aspect, we employed postembedding immunocytochemistry on platelets embedded in the acrylic resin LR White. Clathrin as the major coat component of coated vesicles was localized in the cytoplasm, on the plasmic faces of -granules and the open canalicular system, and on the plasmic face of the plasma membrane. Colocalizations of the adhesive proteins, von Willebrand factor, fibrinogen and fibronectin, with clathrin could be observed at the same typical locations as coated vesicles were seen in Araldite-embedded material. These colocalizations have not been reported to date and furnish further evidence for a coated vesicle-mediated transport of blood plasma-derived adhesive proteins from their receptors on the outer plasma membrane to the -granules.     

Studies on (pro)insulin biosynthesis and secretion of pancreatic islets of sand rats (Psammomys obesus) after different feeding conditions.

It was possible to induce different metabolic states in sand rats of our breeding colony or in newly caught Egyptian sand rats, respectively, by feeding a pellet diet or vegetable diet (green cabbage). Newly captured sand rats fed only on native food were used as reference group (group C). Plasma IRI-level and glucose in vivo and [3H]-leucine incorporation into proinsulin and insulin, insulin secretion and insulin content in vitro were investigated. Sand rats fed on pellet chow and ad libitum (group B) developed a hyperinsulinism and showed higher sensitivity of [3H]-leucine incorporation into proinsulin and insulin to glucose (maximal stimulation at 3 mM) and increased incorporation rates in vitro. Restriction of pellet food to 35-40 kcal/animal/day (group A) lead to changes of all parameters, which were investigated in the same direction as in group B, but to a much smaller extent. Newly captured sand rats, which were fed green cabbage for 4 to 6 weeks divided into two groups: One group (group D1) was comparable to the normal group (C) in IRI levels, glucose levels, glucose sensitivity and amount of [3H]-leucine incorporation. The other group (group D2) tended to group A. Marked changes in insulin content and insulin secretion of isolated pancreatic islets could not be found in any group.     

At least two Toc34 protein import receptors with different specificities are also present in spinach chloroplasts

The receptor components of the chloroplast protein import machinery, Toc34 and Toc159, are both encoded by small gene families in Arabidopsis thaliana. Recent results suggest that each member of these families preferentially interacts with different groups of precursor proteins. Here we address the question, whether multiple homologous Toc receptors are unique to Arabidopsis or whether they are a general phenomenon in plants. Indeed, in spinach we could identify at least two Toc34 proteins with different substrate specificities as demonstrated by competition and antibody inhibition experiments. In addition, an analysis of the available genomic data revealed the presence of at least two Toc34 homologs in six other plant species.     

Radial Turgor and Osmotic Pressure Profiles in Intact and Excised Roots of Aster tripolium: Pressure Probe Measurements and Nuclear Magnetic Resonance-Imaging Analysis

High-resolution nuclear magnetic resonance images (using very short spin-echo times of 3.8 milliseconds) of cross-sections of excised roots of the halophyte Aster tripolium showed radial cell strands separated by air-filled spaces. Radial insertion of the pressure probe (along the cell strands) into roots of intact plants revealed a marked increase of the turgor pressure from the outermost to the sixth cortical layer (from about 0.1-0.6 megapascals). Corresponding measurements of intracellular osmotic pressure in individual cortical cells (by means of a nanoliter osmometer) showed an osmotic pressure gradient of equal magnitude to the turgor pressure. Neither gradient changed significantly when the plants were grown in, or exposed for 1 hour to, media of high salinity. Differences were recorded in the ability of salts and nonelectrolytes to penetrate the apoplast in the root. The reflection coefficients of the cortical cells were approximately 1 for all the solutes tested. Excision of the root from the stem resulted in a collapse of the turgor and osmotic pressure gradients. After about 15 to 30 minutes, the turgor pressure throughout the cortex attained an intermediate (quasistationary) level of about 0.3 megapascals. This value agreed well with the osmotic value deduced from plasmolysis experiments on excised root segments. These and other data provided conclusions about the driving forces for water and solute transport in the roots and about the function of the air-filled radial spaces in water transport. They also showed that excised roots may be artifactual systems.     

Increased organic carbon concentrations in Estonian rivers in the period 1992–2007 as affected by deepening droughts

There are 10,091 km² of peatlands in Estonia and human activities may have changed the role of northern peatlands from global sinks to global sources of carbon. The aim of this work was to explain the changes in organic carbon exports from eleven Estonian rivers in the period 1992–2007 in terms of land use change, climate change quantified by trends in stream-water discharge and hydrological droughts and reductions in atmospheric sulfate quantified by change in water chemistry. Direct TOC (total organic carbon) measurements had been initiated in 1998. We used CODKMnO₄ (permanganate oxygen consumed) as its surrogate for the whole time span (Spearman’s determination coefficient in six small northern Estonian rivers 0.95 > ρ² > 0.72; p < 0.01). The Mann–Kendall test revealed significant trends in the TOC proxy in five small rivers in northern Estonia (M–K stat > 2.35; p < 0.05). The trends in the eleven investigated streams correlated closely with the increased 28 ratios of drought days (Spearman’s ρ² = 0.68; p < 0.01). The correlations with land use compositions, decreases in water discharges and SO₄ ^2? concentrations were insignificant (p > 0.05). We conclude that the main factor in the increase of organic carbon export is the deepening of droughts driven by climate change, magnified by man-made drainage.     

Density-dependent regulation of breeding success in the Red-crested Pochard Netta rufina

Capsule: Numbers of breeding Red-crested Pochard Netta rufina in the Czech Republic were positively correlated with reproductive success in the previous year, yet annual productivity, declined with increasing numbers of females at the beginning of the breeding season, suggesting local limiting factors on this population.

Aims: To test the hypothesis that long-term increases in numbers of a breeding population of Red-crested Pochard Netta rufina can be explained by the productivity (numbers of ducklings fledged) in the previous year. We also test for the potential adverse effect of population size on population productivity in the current year.

Methods: Data from 2004 to 2016 from South Bohemia, Czech Republic, were analysed using TRends and Indices for Monitoring Data software to investigate changes in numbers of adults, numbers of females at the start of the breeding season and number of broods produced.

Results: There was a significant increase in the numbers of adults and females at the beginning of each breeding season. Numbers of adults in May positively correlated with productivity (numbers of ducklings fledged) in the previous breeding season, but relative productivity (broods per adult female) in the current year was negatively correlated with females abundance at the beginning of the breeding season.

Conclusions: The study identifies density-dependent regulation as a factor in the population productivity of Red-crested Pochards using intensively managed fishponds as a breeding ground.     

Acylideneoxoindoles: a new class of reversible inhibitors of human transglutaminase 2

Inhibitors of human transglutaminase 2 (TG2) are anticipated to be useful in the therapy of a variety of diseases including celiac sprue as well as certain CNS disorders and cancers. A class of 3-acylidene-2-oxoindoles was identified as potent reversible inhibitors of human TG2. Structure-activity relationship analysis of a lead compound led to the generation of several potent, competitive inhibitors. Analogs with significant non-competitive character were also identified, suggesting that the compounds bind at one or more allosteric regulatory sites on this multidomain enzyme. The most active compounds had K_i values below 1.0 μM in two different kinetic assays for human TG2, and may therefore be suitable for investigations into the role of TG2 in physiology and disease in animals.