Early changes in adenosine A1 receptors in cerebral cortex slices submitted to in vitro ischemia

The effects of brain ischemia on the maximum binding capacity (B_max) and affinity (K_d) of A₁ receptors were studied in the rat cerebral cortex, with an in vitro approach. The results were correlated with changes in ³H-adenosine release, studied under identical experimental conditions. Fifteen minutes of in vitro ‘ischemia’ (hypoxic, glucose-free medium) induced a significant increase in both B_max (2398±132 fmol/mg protein, 151% of the control, P<0.05) and in K_d (2.43±0.12 nM, 161% of the control, P<0.01). At the same time, an increase in tritium efflux from [³H]-adenosine labeled cerebral cortex slices to 324% of the control was observed. A trend toward normalization was evident 5–15 min after ‘reoxygenation’ (restoring normal medium), but the binding parameters were still altered after 60 min (B_max 2110±82 fmol/mg protein, K_d 2.26±0.14 nM, P<0.01 vs the corresponding control) as was adenosine release (196% of the control). These findings suggest that the increased availability of adenosine and its receptors may be a defense mechanism against ischemic injury, while the reduced affinity of A₁ receptors, possibly due to desensitization, may be a sign of ischemia-induced cellular damage.     

Dietary,Cultural, and Pathogens-Related Selective Pressures Shaped Differential Adaptive Evolution among Native Mexican Populations

Native American genetic ancestry has been remarkably implicated with increased risk of diverse health issues in several Mexican populations, especially in relation to the dramatic changes in environmental, dietary, and cultural settings they have recently undergone. In particular, the effects of these ecological transitions and Westernization of lifestyles have been investigated so far predominantly on Mestizo individuals. Nevertheless, indigenous groups, rather than admixed Mexicans, have plausibly retained the highest proportions of genetic components shaped by natural selection in response to the ancient milieu experienced by Mexican ancestors during their pre-Columbian evolutionary history. These formerly adaptive variants have the potential to represent the genetic determinants of some biological traits that are peculiar to Mexican people, as well as a reservoir of loci with possible biomedical relevance. To test such a hypothesis, we used genome-wide genotype data to infer the unique adaptive evolution of Native Mexican groups selected as reasonable descendants of the main pre-Columbian Mexican civilizations. A combination of haplotype-based and gene-network analyses enabled us to detect genomic signatures ascribable to polygenic adaptive traits plausibly evolved by the main genetic clusters of Mexican indigenous populations to cope with local environmental and/or cultural conditions. Some of these adaptations were found to play a role in modulating the susceptibility/resistance of these groups to certain pathological conditions, thus providing new evidence that diverse selective pressures have contributed to shape the current biological and disease-risk patterns of present-day Native and Mestizo Mexican populations.     

Effect of cadmium, chromium and copper on symbiotic and free-living Rhizobium leguminosarum biovar trifolii

Abstract Plasmid-minus derivatives of Rhizobium leguminosarum biovar trifolii have been isolated. Cured strains lacked symbiotic properties, however they showed increased heavy metal resistance. In the presence of 70 ppm chromium the parent strain, unlike cured derivatives, is unable to grow explanta but can nevertheless nodulate clover.
We propose that rhizobia can circumvent exposure to the heavy metal by entering the plant roots.
Acetylene reduction tests showed that nodulated plants, grown in the presence of 10 ppm of chromium, had an increased nitrogenase activity compared to the control plants.     

Intermittent Anoxia Reduces Oxygen Free Radicals Formation During Reoxygenation in Rat Hepatocytes

The sensitivity of liver cells to anoxia is a major problem afflicting liver preservation and transplantation. Intermittent ischemia has been proposed to reduce reperfusion injury. The aim of the study was to assess oxygen free radical formation and cell injury during continuous or intermittent anoxia/reoxygenation in rat hepatocytes. Anion superoxide was measured by lucigenin-enhanced chemiluminescence and cell damage by LDH release and trypan blue uptake. During anoxia, superoxide generation dropped to background level in both groups; trypan blue uptake and LDH release, which increased progressively, were significantly greater in hepatocytes exposed to continuous compared to intermittent anoxia. During reoxygenation, a massive generation of superoxide anion formation, followed by a sharp increase in LDH release, was observed in both groups. However, both oxyradical generation and cell injury were significantly greater in cells exposed to continuous compared to intermittent anoxia. The data, showing that intermittent oxygen deprivation reduce liver cell injury and oxygen free radical formation determined by anoxia/reoxygenation, suggest a novel possible approach to the reduction of reperfusion injury.     

Human plasma retinol-binding protein (RBP4) is also a fatty acid-binding protein

RBP4 (plasma retinol-binding protein) is the 21?kDa transporter of all-trans retinol that circulates in plasma as a moderately tight 1:1 molar complex of the vitamin with the protein. RBP4 is primarily synthesized in the liver but is also produced by adipose tissue and circulates bound to a larger protein, transthyretin, TTR, that serves to increase its molecular mass and thus avoid its elimination by glomerular filtration.This paper reports the high resolution three-dimensional structures of human RBP4 naturally lacking bound retinol purified from plasma, urine and amniotic fluid. In all these crystals we found a fatty acid molecule bound in the hydrophobic ligand-binding site, a result confirmed by mass spectrometry measurements.In addition we also report the 1.5?Å resolution structures of human holo-RBP4 and of the protein saturated with palmitic and lauric acid and discuss the interaction of the fatty acids and retinol with the protein.     

A Mn(II)–Mn(II) center in human prolidase

Human prolidase, the enzyme responsible for the hydrolysis of the Xaa-Pro/Hyp peptide bonds, is a key player in the recycling of imino acids during the final stage of protein catabolism and extracellular matrix remodeling. Its metal active site composition corresponding to the maximal catalytic activity is still unknown, although prolidase function is of increasing interest due to the link with carcinogenesis and mutations in prolidase gene cause a severe connective tissue disorder. Here, using EPR and ICP-MS on human recombinant prolidase produced in Escherichia coli (hRecProl), the Mn(II) ion organized in a dinuclear Mn(II)–Mn(II) center was identified as the protein cofactor. Furthermore, thermal denaturation, CD/fluorescence spectroscopy and limited proteolysis revealed that the Mn(II) is required for the proper protein folding and that a protein conformational modification is needed in the transition from apo- to Mn(II)loaded-enzyme. The collected data provided a better knowledge of the human holo-prolidase and, although limited to the recombinant enzyme, the exact identity and organization of the metal cofactor as well as the conformational change required for activity were proven.     

Water,land, fire,and forest: Multi‐scale determinants of rainforests in the Australian monsoon tropics

The small rainforest fragments found in savanna landscapes are powerful, yet often overlooked, model systems to understand the controls of these contrasting ecosystems. We analyzed the relative effect of climatic variables on rainforest density at a subcontinental level, and employed high‐resolution, regional‐level analyses to assess the importance of landscape settings and fire activity in determining rainforest density in a frequently burnt Australian savanna landscape. Estimates of rainforest density (ha/km²) across the Northern Territory and Western Australia, derived from preexisting maps, were used to calculate the correlations between rainforest density and climatic variables. A detailed map of the northern Kimberley (Western Australia) rainforests was generated and analyzed to determine the importance of geology and topography in controlling rainforests, and to contrast rainforest density on frequently burnt mainland and nearby islands. In the northwestern Australian, tropics rainforest density was positively correlated with rainfall and moisture index, and negatively correlated with potential evapotranspiration. At a regional scale, rainforests showed preference for complex topographic positions and more fertile geology. Compared with mainland areas, islands had significantly lower fire activity, with no differences between terrain types. They also displayed substantially higher rainforest density, even on level terrain where geomorphological processes do not concentrate nutrients or water. Our multi‐scale approach corroborates previous studies that suggest moist climate, infrequent fires, and geology are important stabilizing factors that allow rainforest fragments to persist in savanna landscapes. These factors need to be incorporated in models to predict the future extent of savannas and rainforests under climate change.     

Relationships between wetland ecotones and inshore water quality in the Ugandan coast of Lake Victoria

Much of the lake shore in Lake Victoria is covered by extensive wetlands, often dominated by dense papyrus stands that extend out over the lake waters. These wetlands, their extension and management play a role in the physical, chemical and biological conditions of the inshore waters. Continuous transects along 180 km of shoreline together with spatial grids of sampling sites in eight bays were performed in the Ugandan inshore waters in order to analyze the relationships between the wetland characteristics and water quality. Measurements of extension of the wetland ecotones, water temperature (T), pH, Secchi disk depth (SD), dissolved oxygen (DO), total nitrogen (TN), total phosphorous (TP), dissolved inorganic nitrogen (DIN), soluble reactive phosphorus (SRP) and chlorophyll-a (CHL) were made in each sampling area. Data of T, pH and DO collected during the transects showed that the water characteristics of the bays differ from the open shoreline. Moreover, the magnitude of these physical–chemical differences is strongly conditioned by the dimension of the bordering wetlands. Bays with extensive wetlands ecotones were characterized by cooler, more acidic and poorly oxygenated waters. TN : TP ratios and especially DIN : SRP ratios decreased with the wetland presence along the coastline, showing a higher probability of N limitation in the inshore waters where large wetlands are present. Results point to denitrification processes in the wetland ecotones as the cause of this trend. The distribution of CHL was found to be highest in the presence of two significant point loading sources: a river (in Katonga Bay) and a major population centre (Kampala, in Murchison Bay). The reduction of external P loading is shown as an important step in the management of the eutrophication process of Lake Victoria inshore waters.