CD8+ T cells are not required for vaccine-induced immunity against Leishmania amazonensis in IL-12/23P40(-/-) C57BL/6 mice

Vaccine-induced protection against leishmaniasis is largely dependent on cell-mediated type 1 response and IL-12-driven IFN-gamma production. Surprisingly, our previous data showed that IL-12/23p40(-/-) mice could be vaccinated against L. amazonensis and were able to produce limited amounts of IFN-gamma. Since the role of CD8+ T in immunization against L. amazonensis is obscure, the aim of this study was to evaluate the effects of CD8+ cells in protection against L. amazonensis in IL-12/23p40(-/-) mice. In order to deplete CD8+ cells, one group of vaccinated animals was treated with anti-CD8 mAb. Infection was followed for 8 weeks. The vaccinated CD8+ -depleted group developed smaller lesions than the non-depleted group. CD8 depletion did not affect tissue parasitism or antibody response against the parasite, and treated animals displayed milder inflammation and better tissue integrity. IFN-gamma production in spleen and draining lymph node was impaired in the depleted group, suggesting that CD8+ cells produced this cytokine in IL-12-independent vaccination. Such results suggest that this T cell subset contributes to augmented pathology in IL12/23p40(-/-) mice vaccinated and challenged with L. amazonensis. Although these cells could produce some IFN-gamma the in the absence of IL-12, they do not affect the parasite tissue load.     

YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7

We have recently characterized Ypi1 as an inhibitory subunit of yeast Glc7 PP1 protein phosphatase. In this work we demonstrate that Ypi1 forms a complex with Glc7 and Sds22, another Glc7 regulatory subunit that targets the phosphatase to substrates involved in cell cycle control. Interestingly, the combination of equimolar amounts of Ypi1 and Sds22 leads to an almost full inhibition of Glc7 activity. Because YPI1 is an essential gene, we have constructed conditional mutants that demonstrate that depletion of Ypi1 leads to alteration of nuclear localization of Glc7 and cell growth arrest in mid-mitosis with aberrant mitotic spindle. These phenotypes mimic those produced upon inactivation of Sds22. The fact that progressive depletion of either Ypi1 or Sds22 resulted in similar physiological phenotypes and that both proteins inhibit the phosphatase activity of Glc7 strongly suggest a common role of these two proteins in regulating Glc7 nuclear localization and function.     

Up‐regulation of SPINT2/HAI‐2 by Azacytidine in bone marrow mesenchymal stromal cells affects leukemic stem cell survival and adhesion

The role of tumour microenvironment in neoplasm initiation and malignant evolution has been increasingly recognized. However, the bone marrow mesenchymal stromal cell (BMMSC) contribution to disease progression remains poorly explored. We previously reported that the expression of serine protease inhibitor kunitz‐type2 (SPINT2/HAI‐2), an inhibitor of hepatocyte growth factor (HGF) activation, is significantly lower in BMMSC from myelodysplastic syndromes (MDS) patients compared to healthy donors (HD). Thus, to investigate whether this loss of expression was due to SPINT2/HAI‐2 methylation, BMMSC from MDS and de novo acute myeloid leukaemia (de novo AML) patients were treated with 5‐Azacitidine (Aza), a DNA methyltransferase inhibitor. In MDS‐ and de novo AML‐BMMSC, Aza treatment resulted in a pronounced SPINT2/HAI‐2 levels up‐regulation. Moreover, Aza treatment of HD‐BMMSC did not improve SPINT2/HAI‐2 levels. To understand the role of SPINT2/HAI‐2 down‐regulation in BMMSC physiology, SPINT2/HAI‐2 expression was inhibited by lentivirus. SPINT2 underexpression resulted in an increased production of HGF by HS‐5 stromal cells and improved survival of CD34⁺ de novo AML cells. We also observed an increased adhesion of de novo AML hematopoietic cells to SPINT2/HAI‐2 silenced cells. Interestingly, BMMSC isolated from MDS and de novo AML patients had increased expression of the integrins CD49b, CD49d, and CD49e. Thus, SPINT2/HAI‐2 may contribute to functional and morphological abnormalities of the microenvironment niche and to stem/progenitor cancer cell progression. Hence, down‐regulation in SPINT2/HAI‐2 gene expression, due to methylation in MDS‐BMMSC and de novo AML‐BMMSC, provides novel insights into the pathogenic role of the leukemic bone marrow microenvironment.     

Inhibition of phospholipase A2 increases Tau phosphorylation at Ser214 in embryonic rat hippocampal neurons

BackgroundArachidonic acid is released from cellular membranes by the action of phospholipase A₂ (PLA₂) and is implicated in microtubule-associated protein Tau phosphorylation. Tau hyperphosphorylation affects its ability to stabilize microtubules.ObjectiveTo determine the effect of PLA₂ inhibition on the phosphorylation state of Tau phosphoepitopes in primary cultures of hippocampal neurons.Methods4 DIC neurons were incubated at different concentrations of methyl-arachidonylfluorophosphonate (MAFP), an irreversible inhibitor of cPLA₂ and iPLA₂. Changes on Tau phosphorylation were determined by Western blotting with a panel of anti-Tau antibodies (C-terminal, Ser199/202, Ser202/205, Ser396 and Ser214).ResultsThe Ser214 site was hyperphosphorylated upon MAFP treatment. Significant differences were observed with 10 μM (p=0.01), 50 μM (p=0.01) and 100 μM (p=0.05) of MAFP. Less-intense changes were found in other phosphoepitopes.ConclusionThe present findings indicate that the phosphorylation of Ser214 is regulated by c- and/or iPLA₂, whereas other phosphoepitopes primarily regulated by GKS3b were not affected.     

Production of an acidic and thermostable lipase of the mesophilic fungus Penicillium simplicissimum by solid-state fermentation

The production of a lipase by a wild-type Brazilian strain of Penicillium simplicissimum in solid-state fermentation of babassu cake, an abundant residue of the oil industry, was studied. The enzyme production reached about 90 U/g in 72 h, with a specific activity of 4.5 U/mg of total proteins. The crude lipase showed high activities at 35–60 °C and pH 4.0–6.0, with a maximum activity at 50 °C and pH 4.0–5.0. Enzyme stability was enhanced at pH 5.0 and 6.0, with a maximum half-life of 5.02 h at 50 °C and pH 5.0. Thus, this lipase shows a thermophilic and thermostable behavior, what is not common among lipases from mesophilic filamentous fungi. The crude enzyme catalysed the hydrolysis of triglycerides and p-nitrophenyl esters (C4:0–C18:0), preferably acting on substrates with medium-chain fatty acids. This non-purified lipase in addition to interesting properties showed a reduced production cost making feasible its applicability in many fields.     

Human adaptations for the visual assessment of strength and fighting ability from the body and face

Selection in species with aggressive social interactions favours the evolution of cognitive mechanisms for assessing physical formidability (fighting ability or resource-holding potential). The ability to accurately assess formidability in conspecifics has been documented in a number of non-human species, but has not been demonstrated in humans. Here, we report tests supporting the hypothesis that the human cognitive architecture includes mechanisms that assess fighting ability-mechanisms that focus on correlates of upper-body strength. Across diverse samples of targets that included US college students, Bolivian horticulturalists and Andean pastoralists, subjects in the US were able to accurately estimate the physical strength of male targets from photos of their bodies and faces. Hierarchical linear modelling shows that subjects were extracting cues of strength that were largely independent of height, weight and age, and that corresponded most strongly to objective measures of upper-body strength-even when the face was all that was available for inspection. Estimates of women's strength were less accurate, but still significant. These studies are the first empirical demonstration that, for humans, judgements of strength and judgements of fighting ability not only track each other, but accurately track actual upper-body strength.     

Lipase production and Penicillium simplicissimum morphology in solid-state and submerged fermentations

A comparative study of Penicillium simplicissimum morphology and lipase production was performed using solid-state (SSF) and submerged (SmF) fermentation. SSF was carried out on babassu cake as culture medium and SmF on a semi-synthetic medium and a medium based on suspended babassu cake grains. Yield of product on biomass, specific activity and conidia production were 3.3-, 1.3- and 2-fold higher in SSF. In SmF, the type of fungus growth differed according to the medium. Using the semi-synthetic medium, the fungus formed densely interwoven mycelial masses without conidia production, whereas using the babassu-based medium the fungus formed free mycelia and adhered to the surfaces of the grains, producing conidia. The results show that babassu cake induces conidiation in SmF. In SSF, the fungus not only grew on the surface of the grains, producing conidia abundantly, but also effectively colonized and penetrated the babassu particles. The high conidia production and lipase productivity in SSF may be related to the low availability of nutrients or to other stimuli associated with this type of fermentation. Thus, the high production of the thermostable P. simplicissimum lipase, using a non-supplemented, low-cost agro-industrial residue as the culture medium, demonstrates the biotechnological potential of SSF for the production of industrial enzymes.     

Control of Dacus oleae by yellow sticky traps combined with ammonium acetate slow-release dispensers

In an isolated olive grove of approximately 350 trees, one yellow sticky trap combined with an ammonium acetate slow-release dispenser was hung in every tree of the large fruit varieties at the end of June or beginning of July for 3 consecutive years. In September, when small fruit varieties were becoming suitable for Dacus oleae (Gmelin) infestation, similar traps were also hung in all trees of these varieties, and the traps in the large fruit trees were replaced by new ones. All traps were removed at the end of December or in January, when harvest of the olive crop was completed. The olive fly population was low in the 1st year, while in the 2nd and 3rd it was high and very high, respectively.In the 1st year, the infestation in the experimental grove was kept very low and was comparable to, or even lower, than the infestation in olive groves treated with insecticides. In the 2nd year, in most cases the infestation was between 15–30%, while in the insecticide-treated groves it remained below 10%. In the 3rd year the infestation reached levels above 80% while in the insecticide-treated groves it reached levels between 30–45%.

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Résumé Un piège gluant jaune associé à un diffuseur libérant lentement de l'acétate d'ammonium, a été suspendu dans chaque arbre d'une variété à gros fruits. L'expérience a eu lieu dans une oliveraie isolée de 350 arbres environ, pendant 3 années consécutives à la fin juin et au début juillet. En septembre, quand les variétés à petits fruits deviennent convenables pour la contamination par D. oleae, des pièges sont suspendus à tous les arbres de ces variétés, les pièges sur les arbres à gros fruits sont alors remplacés. Tous les pièges sont enlevés find décembre ou en janvier, quand la récolte des olives est terminée.En 1982, les températures maximales étaient chaque semaine très élevées de mi-juin à miseptembre (33°C et plus), tandis qu'elles n'étaient atteintes en 1983 et 1984 que de fin juin à mi-août. La population de mouches de l'olive était basse en 1982, tandis qu'elle était élevée en 1983 et très élevée en 1984.La première année (à population faible) l'attaque est restée basse et était comparable ou même plus faible à celle observée dans les oliveraies traitées avec insecticides. La seconde année (population élevée) elle était le plus souvent de 15 à 30% tandis que dans les oliveraies traitées, elle restait inférieure à 10%. La 3è année (population très élevée) l'attaque atteint 80% tandis que dans les oliveraies traitées, elle était de 30 à 45%.