Electron microscopic and histochemical investigations of the atretic oocyte of Perca fluviatilis L. (Teleostei)

Summary The regression of atretic oocytes in Perca fluviatilis was studied by histochemical, light and electron microscopic methods. The course of regression can be divided into three stages, the first two comprising the dissolution of the atretic oocyte and its phagocytosis by the granulosa cells of the follicular epithelium, and the third stage consisting of the dissolution of the granulosa cells themselves. The ultrastructure in all three stages shows only features related to phagocytosis and lysosome formation. In particular, there is no agranular endoplasmic reticulum formed within the phagocytically active granulosa cells, nor is there any 3-hydroxysteroid dehydrogenase activity (3-HSD). Large yellow-orange pigments, formed during the third stage of regression, are ascribed to a relative deficiency of lysosomes in lipid digestion, and do not result from a preceeding steroid-synthesising phase as in mammalian corpora lutea. Thus, the atretic oocyte of P. fluviatilis is considered not to give rise to a corpus luteum formation with endocrine function, but merely represents a degenerative structure.     

Versuche zur Verhinderung von Ultraviolett-Wirkungen auf den Inhalt überlebender tierischer Zellkerne mit Strahlenschutzstoffen

Zusammenfassung Von zehn Substanzen, welche Euglobulin gegen die trübenden und koagulierenden Wirkungen des UV-Lichtes schützen, verhindern drei, nämlich Natriumthioglykolat, NaNO₂ und Na₂SO₃, auch die durch UV erzeugbaren Trübungen und Koagulationen im flüssigen Inhalt von Zellkernen. Dieser Befund wird an zwei verschiedenen überlebenden tierischen Zellen, Eiern der FlußmuschelUnio und Epithelzellen aus der Haut junger Axolotl, erhoben. Wahrscheinlich können nur diese drei Stoffe die Kernmembranen passieren und im Kerninneren auf Grund ihrer reduzierenden Eigenschaften die durch Oxydation bedingten Denaturationen und nachfolgenden Trübungen der Kerneiweiße verhindern.     

Glutamine Transport in Rat Brain Synaptic and Non-Synaptic Mitochondria

Glutamine transport into rat brain mitochondria (synaptic and non-synaptic) was monitored by the uptake of [³H]glutamine as well as by mitochondrial swelling. The uptake is inversely correlated to medium osmolarity, temperature-dependent, saturable and inhibited by mersalyl, and glutamine is upconcentrated in the mitochondria. These results indicate that glutamine is transported into an osmotically active space by a protein catalyzed mechanism. The uptake is slightly higher in synaptic mitochondria than in non-synaptic ones. It is inhibited both by rotenone and the protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone, the latter at pH 6.5, showing that the transport is activated by an electrochemical proton gradient. The K⁺/H⁺ ionophore nigericin also inhibits the uptake at pH 6.5 in the presence of external K⁺, which indicates that glutamine, at least in part, is taken up by a proton symport transporter. In addition, glutamine uptake as measured by the swelling technique revealed an additional glutamine transport activity with at least 10 times higher K_m value. This uptake is inhibited by valinomycin in the presence of K⁺ and is thus also activated by the membrane potential. Otherwise, the two methods show similar results. These data indicate that glutamine transport in brain mitochondria cannot be described by merely a simple electroneutral uniport mechanism, but are consistent with the uptake of both the anionic and the zwitterionic glutamine.     

Comparison of [3H]WIN 35,428 Binding, a Marker for Dopamine Transporter, in Embryonic Mesencephalic Neuronal Cultures with Striatal Membranes of Adult Rats

Abstract: In contrast to striatal membranes of adult rats, where high- ( K _D1= 34 n M ) and low- ( K _D2= 48,400 n M ) affinity binding sites for [³H]WIN 35,428 are present, in primary cultures of ventral mesencephalon neurons (CVMNs) only low-affinity binding sites were found ( K _D= 336,000 n M ). The binding of [³H]WIN 35,428 in CVMNs prepared from rat embryos was reversible, saturable, and located in cytosol. Although dopamine (DA) uptake blockers inhibited [³H]DA uptake at nanomolar concentrations in CVMNs, the displacement of [³H]WIN 35,428 binding in CVMNs by DA uptake inhibitors required 100-8,000 times higher concentrations than were needed to displace [³H]WIN 35,428 binding in striatal membranes. Piperazine derivatives, e.g., GBR-12909, GBR-12935, and rimcazole, inhibited [³H]WIN 35,428 binding in CVMNs more effectively than did cocaine, WIN 35,428, mazindol, nomifensine, or benztropin. A positive correlation ( r = 0.779; p < 0.001) was found between drug affinities for the striatal membrane sites labeled by [³H]WIN 35,428 and their abilities to inhibit DA uptake in CVMNs, whereas no correlation existed between the IC₅₀ values of drugs that inhibited [³H]WIN 35,428 binding and [³H]DA uptake in CVMNs. The cytosolic [³H]WIN 35,428 binding sites may be a piperazine acceptor and may not be involved in the regulation of the DA transporter.     

Structures de végétation et conservation des zones humides temporaires méditerranéennes : la région des Mogods (Tunisie septentrionale)

Floristic surveys and phytoecological relevés were conducted on 36 temporary wetlands of Mogods region. Multivariate analyses (CA, AHC) performed on these data reveal the high specific and biocoenotic diversity of Mogods wetlands, which appear controlled by substrate nature and hydrology. Among the 128 hydrophytic species inventoried, 38 are presently in precarious status and 6 are presumed extinct. The Mogods region harbours, moreover, very rare habitats (peatlands and semi-permanent lakes), and a vast plain so-called Garâa Sejenane, exceptionally rich in temporary wetlands. These results underline the urgency of an adapted conservatory management, based on the development of scientific studies dealing with structure and functioning of hydrophytic communities of the region.     

Identification of potential serum biomarkers of inflammation and lipid modulation that are altered by fish oil supplementation in healthy volunteers

Long chain n-3 polyunsaturated fatty acids (n-3 LCPUFA) lower risk of coronary heart disease (CHD), but mechanisms are not well understood. We used proteomics to identify human serum proteins that are altered by n-3 LCPUFA. Such proteins could identify pathways whereby they affect CHD. Eighty-one healthy volunteers entered a double blind randomised trial to receive 3.5 g of fish oil or 3.5 g of high oleic sunflower oil daily. Serum was collected before and after 6 wk of intervention. Serum was analysed by proteomics using 2-DE. Proteins that were differentially regulated were identified by MS. We also analysed serum apolipoprotein A1 (apo A1), high-density lipoprotein (HDL) particle size and haptoglobin. Serum levels of apo A1, apo L1, zinc-alpha-2-glycoprotein, haptoglobin precursor, alpha-1-antitrypsin precursor, antithrombin III-like protein, serum amyloid P component and haemopexin were significantly downregulated (all p<0.05) by fish oil compared with high oleic sunflower oil supplementation. Fish oil supplementation caused a significant shift towards the larger, more cholesterol-rich HDL(2) particle. The alterations in serum proteins and HDL size imply that fish oil activates anti-inflammatory and lipid modulating mechanisms believed to impede the early onset of CHD. These proteins are potential diagnostic biomarkers to assess the mechanisms whereby fish oil protects against CHD in humans.     

Throughfall Nitrogen Deposition Has Different Impacts on Soil Solution Nitrate Concentration in European Coniferous and Deciduous Forests

Increases in the deposition of atmospheric nitrogen (N) influence N cycling in forest ecosystems and can result in negative consequences due to the leaching of nitrate into groundwaters. From December 1995 to February 1998, the Pan-European Programme for the Intensive and Continuous Monitoring of Forest Ecosystems measured forest conditions at a plot scale for conifer and broadleaf forests, including the performance of time series of soil solution chemistry. The influence of various ecosystem conditions on soil solution nitrate concentrations at these forest plots (n = 104) was then analyzed with a statistical model. Soil solution nitrate concentrations varied by season, and summer concentrations were approximately 25% higher than winter ones. Soil solution nitrate concentrations increased dramatically with throughfall (and bulk precipitation) N input for both broadleaf and conifer forests. However, at elevated levels of throughfall N input (more than 10 kg N ha^–1 y^–1), nitrate concentrations were higher in broadleaf than coniferous stands. This tree-specific difference was not observed in response to increased bulk precipitation N input. In coniferous stands, throughfall N input, foliage N concentration, organic layer carbon–nitrogen (C:N) ratio, and nitrate concentrations covaried. Soil solution nitrate concentrations in conifer plots were best explained by a model with throughfall N and organic layer C:N as main factors, where C:N ratio could be replaced by foliage N. The organic layer C:N ratio classes of more than 30, 25–30, and less than 25, as well as the foliage N (mg N g^–1) classes of less than 13, 13–17, and more than 17, indicated low, intermediate, and high risks of nitrate leaching, respectively. In broadleaf forests, correlations between N characteristics were less pronounced, and soil solution nitrate concentrations were best explained by throughfall N and soil pH (0–10-cm depth). These results indicate that the responses of soil solution nitrate concentration to changes in N input are more pronounced in broadleaf than in coniferous forests, because in European forests broadleaf species grow on the more fertile soils.     

The angio-fibrotic switch of VEGF and CTGF in proliferative diabetic retinopathy

Background

In proliferative diabetic retinopathy (PDR), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) cause blindness by neovascularization and subsequent fibrosis, but their relative contribution to both processes is unknown. We hypothesize that the balance between levels of pro-angiogenic VEGF and pro-fibrotic CTGF regulates angiogenesis, the angio-fibrotic switch, and the resulting fibrosis and scarring.

Methods/Principal Findings

VEGF and CTGF were measured by ELISA in 68 vitreous samples of patients with proliferative DR (PDR, N = 32), macular hole (N = 13) or macular pucker (N = 23) and were related to clinical data, including degree of intra-ocular neovascularization and fibrosis. In addition, clinical cases of PDR (n = 4) were studied before and after pan-retinal photocoagulation and intra-vitreal injections with bevacizumab, an antibody against VEGF. Neovascularization and fibrosis in various degrees occurred almost exclusively in PDR patients. In PDR patients, vitreous CTGF levels were significantly associated with degree of fibrosis and with VEGF levels, but not with neovascularization, whereas VEGF levels were associated only with neovascularization. The ratio of CTGF and VEGF was the strongest predictor of degree of fibrosis. As predicted by these findings, patients with PDR demonstrated a temporary increase in intra-ocular fibrosis after anti-VEGF treatment or laser treatment.

Conclusions/Significance

CTGF is primarily a pro-fibrotic factor in the eye, and a shift in the balance between CTGF and VEGF is associated with the switch from angiogenesis to fibrosis in proliferative retinopathy.     

Synergistic induction of osteopontin by aldosterone and inflammatory cytokines in mesangial cells

Hypertensive nephrosclerosis is characterized by activation of the renin-angiotensin-aldosterone system in combination with an inflammatory response characterized by an infiltration of T-cells and mononuclear cells, which release proinflammatory cytokines like IL-1beta/TNFalpha. In various models of experimental hypertensive disease the chemokine osteopontin (OPN) enhances further leukocyte infiltration. Therefore, we investigated the induction of OPN expression in renal mesangial cells (MCs) by aldosterone and the inflammatory cytokines IL-1beta/TNFalpha. Incubation with aldosterone resulted in a time- and concentration-dependent increase in OPN mRNA and protein. OPN mRNA expression followed a biphasic time course with an early increase between 4 and 8 h and the second phase starting at 14 h. The early phase was independent of protein synthesis, indicating a direct effect of aldosterone. Aldosterone-mediated induction of OPN was prevented by spironolactone, indicative of a receptor-mediated aldosterone effect. The mineralocorticoid receptor (MR) was identified in MCs by RT-PCR and immunoprecipitation, and shown to interact with a putative aldosterone-response element of the OPN promoter. The proinflammatory cytokines IL-1beta and TNFalpha only marginally affected OPN expression in MCs. However, coincubation of aldosterone and the cytokines synergistically increased OPN mRNA and protein levels. Since the synergistic effect on OPN mRNA was inhibited by diphenyleneiodonium, we assume an involvement of reactive oxygen species (ROS). We conclude that the chemokine OPN is a target gene of aldosterone in renal MCs, which is activated via the MR, and that proinflammatory cytokines enhance aldosterone-dependent OPN expression. In vivo, this may result in further leukocyte infiltration aggravating hypertensive nephrosclerosis.