IL-10 and TGF-beta control the establishment of persistent and transmissible infections produced by Leishmania tropica in C57BL/6 mice

Leishmania tropica is the causative agent of Old World anthroponotic cutaneous leishmaniasis, which is characterized by lesions that take an extended period of time to heal, often resulting in disfiguring scars, and are more refractory to treatment than leishmaniasis caused by Leishmania major. Immunologic studies involving experimental animal models of L. tropica infection are virtually nonexistent. In the current study, infectious-stage L. tropica were used to establish dermal infections in C57BL/6 and BALB/c mice. In both strains, the lesions were slow to develop and showed minimal pathology. They nonetheless contained a stable number of between 10(4) and 10(5) parasites for over 1 year, which were efficiently picked up by a natural sand fly vector, Phlebotomus sergenti. Control of parasite growth depended on the development of a Th1 response, as C57BL/6 mice genetically deficient in Th1 cytokines and BALB/c mice treated with Abs to IFN-gamma harbored significantly more parasites. By contrast, IL-10-deficient mice harbored significantly fewer parasites throughout the infection. To further study the immunologic mechanisms that may prevent efficient clearance of the parasites, IL-10 and TGF-beta signaling were abrogated during the chronic phase of infection in wild-type C57BL/6 mice. Distinct from chronic L. major infection, IL-10 blockade alone had no effect on L. tropica, but required simultaneous treatment with anti-TGF-beta Abs to promote efficient parasite clearance from the infection site. Thus, chronic infection with L. tropica appears to be established via multiple suppressive factors, which together maintain the host as a long-term reservoir of infection for vector sand flies.     

Effect of rhizobia and rock biofertilizers with <Emphasis Type="Italic">Acidithiobacillus</Emphasis> on cowpea nodulation and nutrients uptake in a tableland soil

Chemical fertilizers have been used in the cultivation of plants due to their high solubility and effect on crops yield. Biofertilizers with phosphate rock (PR) and potash rock (KR) plus sulfur inoculated with Acidithiobacillus may improve plant growth and contribute to addition of available P and K in soil. The effectiveness of biofertilizers from phosphate and potash rocks mixed with sulfur and Acidithiobacillus was studied in a Typic Fragiuldult soil of the Brazilian Northeast Tableland. Cowpea (cv. “IPA 206”) was grown with and without rhizobia inoculation. Treatments were: (a) phosphate rock (1000 kg ha⁻¹); (b) Biofertilizers-B_P (250 and 500 kg ha⁻¹); (c) triple superphosphate-TSP (250 kg ha⁻¹); (d) potash rock (1000 kg ha⁻¹); (e) biofertilizer-B_K (250; 500 and 750 kg ha⁻¹); (f) potassium chloride-KCl (250 kg K₂0 ha⁻¹); (g) control without P or K fertilization (P₀K₀). The soil was maintained under water submersion covered with black plastic (solarization process) for a period of 30 days. Biofertilizers (B_p and B_K) and soluble fertilizers increased plant growth and NPK uptake. Biofertilizers reduced soil pH, especially when applied in highest rates. Biofertilizers and TSP+KCl showed the best values of available P and K in soil. Rhizobial inoculation was effective on cowpea, but no nodules were formed by bacteria native from the soil, probably due to the effect of the solarization process. From obtained PK biofertilizers could be used as alternative for cowpea fertilization in Tableland soils.     

Genetik und Klinik der Hämophilie A und B

Haemophilia A and B are caused by various mutations in the factor VIII (FVIII) and factor IX (FIX) genes, respectively. The clinical course of the disease is variable, dependent on the severity of the molecular defect. Nowadays, haemophilia patients can excellently be treated by plasma-derived or recombinant clotting factor concentrates. Thus, bleeding and its consequences can be almost completely prevented with nearly normal quality of life and life expectancy. The most severe complication of this treatment is the formation of antibodies (inhibitors) against the substituted clotting factor. The risk of inhibitor formation correlates significantly with specific mutation types that preclude endogenous factor VIII/IX protein synthesis and can be as high as 20–50%. The information on the expected clinical course is at present the most important indication for FVIII/IX gene analysis. Knowledge of the underlying FVIII/IX gene mutation further allows a reliable and fast carrier diagnosis in female relatives of patients with haemophilia.     

Expression in Escherichia coli, purification, refolding and antifungal activity of an osmotin from Solanum nigrum

Background

The large-scale production of G-protein coupled receptors (GPCRs) for functional and structural studies remains a challenge. Recent successes have been made in the expression of a range of GPCRs using Pichia pastoris as an expression host. P. pastoris has a number of advantages over other expression systems including ability to post-translationally modify expressed proteins, relative low cost for production and ability to grow to very high cell densities. Several previous studies have described the expression of GPCRs in P. pastoris using shaker flasks, which allow culturing of small volumes (500 ml) with moderate cell densities (OD600 ~15). The use of bioreactors, which allow straightforward culturing of large volumes, together with optimal control of growth parameters including pH and dissolved oxygen to maximise cell densities and expression of the target receptors, are an attractive alternative. The aim of this study was to compare the levels of expression of the human Adenosine 2A receptor (A_2AR) in P. pastoris under control of a methanol-inducible promoter in both flask and bioreactor cultures.

Results

Bioreactor cultures yielded an approximately five times increase in cell density (OD600 ~75) compared to flask cultures prior to induction and a doubling in functional expression level per mg of membrane protein, representing a significant optimisation. Furthermore, analysis of a C-terminally truncated A_2AR, terminating at residue V334 yielded the highest levels (200 pmol/mg) so far reported for expression of this receptor in P. pastoris. This truncated form of the receptor was also revealed to be resistant to C-terminal degradation in contrast to the WT A_2AR, and therefore more suitable for further functional and structural studies.

Conclusion

Large-scale expression of the A_2AR in P. pastoris bioreactor cultures results in significant increases in functional expression compared to traditional flask cultures.     

Reaching the canopy on the ground: incidence of infection and host-use by mistletoes (Loranthaceae and Viscaceae) on trees felled for timber in Amazonian rainforests

There is a profound absence of knowledge of infestation prevalence and host-use by mistletoes of mature South American tropical rainforests. In this study, we fill this gap using information gathered from felled trees at a logging concession area in Amazonian Brazil. We sampled individuals of 18 tree species, which occurred in two forest physiognomies; open forest with canopy interrupted by palm trees and closed, denser forest, with emergent trees. We hypothesized that infection incidence would be higher in open than in closed forest, irrespective of the mistletoe species involved. In addition, we expected that mistletoe parasitism would be higher on host species that were more abundant, taller, deciduous, and had less dense wood. We sampled 870 individual trees in both sites combined. All but one host species was infected by at least one species of mistletoe. We found 13 mistletoe species/morphospecies, Loranthaceae (7) and Viscaceae (6), parasitizing very different hosts. Mistletoe infection incidence was higher in the closed forest (10.3%) than in the open forest (5.4%). In the closed forest, host height influenced incidence positively, while deciduousness had a negative influence. Our results show that mistletoes are common in the canopy of pristine tropical forests and, contrary to expectations, that infection incidence was higher in the closed forest. The positive relation between infection incidence and host height in this forest type suggests that emergent trees have higher chances of being infected than individuals of correspondent species in the lower forest layers.     

A proposal for the conservation of the family Cucurbitaceae in Mexico

The pantropical Cucurbitaceae is one of the most important families ofvascular plants. The family includes 118 genera and 825 species, and Mexicois one of its most important centers of diversity, with 34 genera and 141species and subspecific taxa, including 13 cultivated ones. Five genera and 70taxa are endemic to the country. Some Mexican and Latin American wild speciesare close relatives of important crops, and others are employed as food ormedicine. Accordingly, in countries like Mexico the conservation of members ofthis family should be a priority. In this paper a list of members ofCucurbitaceae occurring in Mexico and their distribution patterns at state levelare discussed. Using cluster strategies, the states were classified according totheir floristic similarities. Hotspots of total diversity and endemismsat state level are identified, in order to discuss their role in futureconservation strategies. Iterative methods applied by conservation biology todetermine the best places for conservation are used to identify hierarchicallythe most important states that merit to be considered in this goal. Resultsindicate that implementing conservation strategies in half of the states wouldallow the protection of all the diversity found in the country, especially theendemisms. The combination of the results provided by the different methods isalso discussed as a possibly more efficient way to propose conservationstrategies for important species of the family.     

Plant resources of the Tehuacán-Cuicatlán valley, Mexico

Information on richness of plant resources, and their forms of use and management in the biosphere reserve Tehuacan-Cuicatlan, Mexico is analyzed. This 10 000 km² region hosts nearly 2700 vascular plant species, and it is acknowledged as one of the arid areas with the highest floristic diversity in North America. The seven indigenous ethnic groups that live in this region have cultural roots that date back almost 10 000 years. Based upon ethnobotanical and floristic studies, as well as bibliographical sources, a total of 808 useful plant species were identified, most of them (90%) being native, and 44 species being endemic to the region. A total of 681 species are wild plants, 109 are weeds and ruderal plants, and 86 are domesticated crops. However, it was noted that considerable overlap exists between the species of these 3 categories. For example, while wild and ruderal plants (706 species) are foraged by both humans and domestic animals, 59 species of this group are also managed in situ. On the other hand, 168 wild, ruderal and domesticated species are cultivated. The Tehuacán-Cuicatlan Valley is one of the richest regions of Mexico in plant resources. Local knowledge on use and management of plants is a valuable source of information for designing conservation and social development strategies for the biosphere reserve.     

Yerba mate extract (Ilex paraguariensis) attenuates both central and peripheral inflammatory effects of diet-induced obesity in rats

To clarify the effects of natural dietary components on the metabolic consequences of obesity, we examined the effects of yerba mate extract Ilex paraguariensis on both central and peripheral inflammatory effects of diet-induced obesity and correlated the hypothalamic tumor necrosis factor (TNF)-α level with adipose depot weight. Wistar rats were divided into four groups: a control group (CTL) fed with chow diet, a second group fed with chow diet plus yerba mate extract (CTL+E), a third group fed with a high-fat diet rich in saturated fatty acids (HFD) and a fourth group fed with HFD plus yerba mate extract (HFD+E). Enzyme-linked immunosorbent assay, Western blotting, colorimetric method and treatment by gavage were utilized as materials and methods. The HFD groups showed a significant increase in food intake (kcal), body weight, adipose tissue and leptin level in comparison to CTL and CTL+E. HFD leads to increase of both central and peripheral inflammatory effects, and deregulation of insulin pathway. In addition, yerba mate extract intake blunted the proinflammatory effects of diet-induced obesity in rats by reducing the phosphorylation of hypothalamic IKK and NFκBp65 expression and increasing the protein levels of IκBα, the expression of adiponectin receptor-1 and consequently the amount of IRS-2. Moreover, the increase in interleukin (IL)-6 levels in the liver and muscle and of the IL-10/TNF-α ratio in groups that received yerba mate extract showed the anti-inflammatory effects of this natural substance. Taken together, our data suggest that the use of yerba mate extract may be useful for reducing low-grade obesity-associated inflammation.