Polinização de pepino, Cucumis sativus L. (Cucurbitales: Cucurbitaceae), pelas abelhas sem ferrão Scaptotrigona aff. depilis moure e Nannotrigona testaceicornis lepeletier (Hymenoptera: Meliponini) em casas de vegetação

When for a successful fruit development the fertilization of flowers is necessary, bees can be used as crop-pollinators in greenhouses. In the present study, we investigated the effectiveness of the stingless bees Scaptotrigona aff. depilis Moure and Nannotrigona testaceicornis Lepeletier as pollinators of cucumber plants (Cucumus sativus var. caipira) in greenhouses during the Brazilian winter season. The study was conducted in four greenhouses (GH), of which two greenhouses contained bee colonies to ascertain pollination of the cucumber plants (GH I, with S. aff. depilis, GH II, with N. testaceicornis), whereas the other two greenhouses (GH III, GH IV) had no bee colonies and served as control groups. Furthermore, we planted cucumbers in an open field plot (OA) where pollination by any/various visiting insects could occur. Each of the experimental areas measured 87.5 m². Without pollination (GH III, GH IV), the plants produced a low number of cucumbers, and the fruits were smaller and less heavy than in those experimental areas where pollination occurred. In the open field area, not protected against unfavorable climatic conditions, the plants produced fewer flowers than the plants in the greenhouses. The highest cucumber yield (with the highest amount of perfect fruits) was found in those greenhouses which housed the stingless bees as pollinators (GH I, GH II). Our results demonstrate that stingless bees can be successfully and efficiently used as pollinators of greenhouse cucumbers during the winter season.Quando a fertilização de flores é necessária para o desenvolvimento de frutos, abelhas podem ser utilizadas como polinizadores sob cultivo protegido. No presente estudo, a efetividade das abelhas sem ferrão Scaptotrigona aff. depilis Moure e Nannotrigona testaceicornis Lepeletier como polinizadoras de pepino (Cucumis sativus var. caipira) foi investigada sob cultivo protegido durante o inverno. O estudo foi conduzido em quatro casas de vegetação (GH), das quais duas continham colônias de abelhas para averiguar a polinização dos pepinos (GH I, com S. aff. depilis, GH II, com N. testaceicornis), e duas (GH III, GH IV) não continham colônias e serviram como grupos controle. Além disso, pepinos foram plantados numa área aberta (AO) onde polinização por vários insetos poderia ocorrer. Sem polinização (GH III, GH IV), as plantas produziram menor número de pepinos, e os frutos eram menores e menos pesados do que aqueles nas áreas experimentais onde a polinizacão ocorreu. Na área aberta, não protegida contra condições climáticas desfavoráveis, as plantas produziram menos flores do que as plantas nas casas de vegetação. A maior produção de pepinos (com a maior quantidade de frutos perfeitos) foi encontrada nas casas de vegetação com as abelhas como polinizadoras (GH I, GH II). Os resultados demonstraram que abelhas sem ferrão podem ser usadas com sucesso e eficiência como polinizadoras de pepinos sob cultivo protegido durante o inverno.FAPESPCoordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)CNP

Immunomodulatory effects of palladium(II) complexes of 1,2,4-triazole on murine peritoneal macrophages

The 1,2,4-triazolyl-bridged polynuclear complexes [{PdCl(2)(mu-Htrz)}(n)] (1) and [{PdBr(2)(mu-Htrz)}(n)] (2) have been obtained in this work. Compound 1 is prepared by the displacement of acetonitrile from [PdCl(2)(MeCN)(2)] by 1,2,4-triazole (Htrz). Further addition of potassium bromide to the reaction medium afforded complex 2. The new complexes have been isolated, purified and characterized by means of elemental analysis, IR and UV-visible electronic spectroscopies and thermogravimetric (TG) curves. The experimental data suggested that, in both cases, the coordination of 1,2,4-Htrz takes place through the N(2) and N(4) atoms, bridging the palladium centers. The square-planar coordination polyhedron of palladium(II) is determined by two nitrogen atoms from the triazole ligands, while the other two coordination positions are occupied by the chloro (1) or bromo (2) ligands. TG curves indicated that the nature of the anionic ligand does not affect significantly the thermal stability of 1 and 2. The final products of the thermal decompositions were identified as metallic palladium by X-ray powder diffractometry. Preliminary tests involving the evaluation of the effects of compounds 1, 2 and Htrz on H(2)O(2) and NO production in cultures of peritoneal macrophages from BALB/c mice were carried out in vitro

Bee communities (Hymenoptera: Anthophila) of the "Cerrado" ecosystem in Sao Paulo State, Brazil

Five surveys of the bee communities in four "Cerrado" ecosystem reserves in Sao Paulo State were compared for species richness and similarity. These areas are fragment vegetation reser-reserves located in the Cerrado Corumbata Reserve (Corumbata), Jata Ecological Park (Luiz Antonio), Cajuru (Cajuru), and Vassununga State Park - ""Gleba de Cerrado de Pe-de-Gigante"" (Santa Rita do Passa Quatro). The methodology consisted of capturing bees foraging on flowers along transects, though with small differences between surveys. These ""cerrado"" areas have a large number of species of native bees, which are important pollinators in several Brazilian ecosystems. The community of bees varied among these different fragments. Based on 500 individuals (standardized by rarefaction), Cajuru, Corumbata 1 and Corumbata 2 were the areas with highest species richness, and Jata and Pe-de-Gigante had the lowest species richness in the bee communities. The bee faunas of Corumbata 2 and Pe-de-Gigante had the highest similarity, forming a group with the bee fauna of Cajuru. The bee faunas of Corumbata 1 and Jata were isolated from this group. We found that the bee species richness and similarity found in these ""cerrado"" areas cannot be explained by general factors such as the size of the fragment, the species richness of plants and the distance between the areas. Therefore, we suppose that local factors that differ among areas, such as interactions between populations, and competition and interference from surrounding areas influence and determine bee species richness and similarity in these reserves.FAPESP (Funda ao de Amparo a Pesquisa do Estado de Sao Paulo)[00/06405-2]CNP

Genetical and comparative genomics of Brassica under altered Ca supply identifies Arabidopsis Ca-transporter orthologs

Although Ca transport in plants is highly complex, the overexpression of vacuolar Ca2+ transporters in crops is a promising new technology to improve dietary Ca supplies through biofortification. Here, we sought to identify novel targets for increasing plant Ca accumulation using genetical and comparative genomics. Expression quantitative trait locus (eQTL) mapping to 1895 cis- and 8015 trans-loci were identified in shoots of an inbred mapping population of Brassica rapa (IMB211 x R500); 23 cis- and 948 trans-eQTLs responded specifically to altered Ca supply. eQTLs were screened for functional significance using a large database of shoot Ca concentration phenotypes of Arabidopsis thaliana. From 31 Arabidopsis gene identifiers tagged to robust shoot Ca concentration phenotypes, 21 mapped to 27 B. rapa eQTLs, including orthologs of the Ca2+ transporters At-CAX1 and At-ACA8. Two of three independent missense mutants of BraA.cax1a, isolated previously by targeting induced local lesions in genomes, have allele-specific shoot Ca concentration phenotypes compared with their segregating wild types. BraA.CAX1a is a promising target for altering the Ca composition of Brassica, consistent with prior knowledge from Arabidopsis. We conclude that multiple-environment eQTL analysis of complex crop genomes combined with comparative genomics is a powerful technique for novel gene identification/prioritization

KCNQ1 suppression-replacement gene therapy in transgenic rabbits with type 1 long QT syndrome.

BACKGROUND AND AIMS Type 1 long QT syndrome (LQT1) is caused by pathogenic variants in the KCNQ1-encoded Kv7.1 potassium channels, which pathologically prolong ventricular action potential duration (APD). Herein, the pathologic phenotype in transgenic LQT1 rabbits is rescued using a novel KCNQ1 suppression-replacement (SupRep) gene therapy. METHODS KCNQ1-SupRep gene therapy was developed by combining into a single construct a KCNQ1 shRNA (suppression) and an shRNA-immune KCNQ1 cDNA (replacement), packaged into adeno-associated virus serotype 9, and delivered in vivo via an intra-aortic root injection (1E10 vg/kg). To ascertain the efficacy of SupRep, 12-lead electrocardiograms were assessed in adult LQT1 and wild-type (WT) rabbits and patch-clamp experiments were performed on isolated ventricular cardiomyocytes. RESULTS KCNQ1-SupRep treatment of LQT1 rabbits resulted in significant shortening of the pathologically prolonged QT index (QTi) towards WT levels. Ventricular cardiomyocytes isolated from treated LQT1 rabbits demonstrated pronounced shortening of APD compared to LQT1 controls, leading to levels similar to WT (LQT1-UT vs. LQT1-SupRep, P < .0001, LQT1-SupRep vs. WT, P = ns). Under β-adrenergic stimulation with isoproterenol, SupRep-treated rabbits demonstrated a WT-like physiological QTi and APD90 behaviour. CONCLUSIONS This study provides the first animal-model, proof-of-concept gene therapy for correction of LQT1. In LQT1 rabbits, treatment with KCNQ1-SupRep gene therapy normalized the clinical QTi and cellular APD90 to near WT levels both at baseline and after isoproterenol. If similar QT/APD correction can be achieved with intravenous administration of KCNQ1-SupRep gene therapy in LQT1 rabbits, these encouraging data should compel continued development of this gene therapy for patients with LQT1

Abelhas-sem-ferrão amazônicas defendem meliponários contra saques de outras abelhas

Entre as abelhas eussociais, dois gêneros apresentam estratégia de vida cleptobiótica, obtendo recursos alimentares de ninhos de outras abelhas ao invés de coletá-los em flores. Entre as espécies atacadas existe um gradiente de suscetibilidade ao roubo variando desde espécies vulneráveis até altamente resistentes. Neste trabalho nós descrevemos um ataque de Lestrimelitta rufipes a um ninho de Scaptotrigona sp. em um meliponário na Amazônia central (Amazonas, Brazil). O ninho atacado foi transferido para um meliponário com espécies resistentes (Duckeola ghilianii e Melipona fulva) e as interações foram descritas. As abelhas resistentes contra-atacaram e afugentaram as ladras protegendo o ninho de Scaptotrigona sp.. A presença de comportamento defensivo em gêneros não proximamente relacionados sugere que ele tenha evoluído mais de uma vez entre os Meliponini. Considerando o comportamento descrito, sugerimos a criação de espécies nativas resistentes em meliponários de regiões onde elas forem nativas, devido ao potencial que elas tem na proteção

Human cytomegalovirus latency-associated proteins elicit immune-suppressive IL-10 producing CD4⁺ T cells.

Human cytomegalovirus (HCMV) is a widely prevalent human herpesvirus, which, after primary infection, persists in the host for life. In healthy individuals, the virus is well controlled by the HCMV-specific T cell response. A key feature of this persistence, in the face of a normally robust host immune response, is the establishment of viral latency. In contrast to lytic infection, which is characterised by extensive viral gene expression and virus production, long-term latency in cells of the myeloid lineage is characterised by highly restricted expression of viral genes, including UL138 and LUNA. Here we report that both UL138 and LUNA-specific T cells were detectable directly ex vivo in healthy HCMV seropositive subjects and that this response is principally CD4⁺ T cell mediated. These UL138-specific CD4⁺ T cells are able to mediate MHC class II restricted cytotoxicity and, importantly, show IFNγ effector function in the context of both lytic and latent infection. Furthermore, in contrast to CDCD4⁺ T cells specific to antigens expressed solely during lytic infection, both the UL138 and LUNA-specific CD4⁺ T cell responses included CD4⁺ T cells that secreted the immunosuppressive cytokine cIL-10. We also show that cIL-10 expressing CD4⁺ T-cells are directed against latently expressed US28 and UL111A. Taken together, our data show that latency-associated gene products of HCMV generate CD4⁺ T cell responses in vivo, which are able to elicit effector function in response to both lytic and latently infected cells. Importantly and in contrast to CD4⁺ T cell populations, which recognise antigens solely expressed during lytic infection, include a subset of cells that secrete the immunosuppressive cytokine cIL-10. This suggests that HCMV skews the T cell responses to latency-associated antigens to one that is overall suppressive in order to sustain latent carriage in vivo

Cyclin-Dependent Kinase Activity Controls the Onset of the HCMV Lytic Cycle

The onset of human cytomegalovirus (HCMV) lytic infection is strictly synchronized with the host cell cycle. Infected G0/G1 cells support viral immediate early (IE) gene expression and proceed to the G1/S boundary where they finally arrest. In contrast, S/G2 cells can be infected but effectively block IE gene expression and this inhibition is not relieved until host cells have divided and reentered G1. During latent infection IE gene expression is also inhibited, and for reactivation to occur this block to IE gene expression must be overcome. It is only poorly understood which viral and/or cellular activities maintain the block to cell cycle or latency-associated viral IE gene repression and whether the two mechanisms may be linked. Here, we show that the block to IE gene expression during S and G2 phase can be overcome by both genotoxic stress and chemical inhibitors of cellular DNA replication, pointing to the involvement of checkpoint-dependent signaling pathways in controlling IE gene repression. Checkpoint-dependent rescue of IE expression strictly requires p53 and in the absence of checkpoint activation is mimicked by proteasomal inhibition in a p53 dependent manner. Requirement for the cyclin dependent kinase (CDK) inhibitor p21 downstream of p53 suggests a pivotal role for CDKs in controlling IE gene repression in S/G2 and treatment of S/G2 cells with the CDK inhibitor roscovitine alleviates IE repression independently of p53. Importantly, CDK inhibiton also overcomes the block to IE expression during quiescent infection of NTera2 (NT2) cells. Thus, a timely block to CDK activity not only secures phase specificity of the cell cycle dependent HCMV IE gene expression program, but in addition plays a hitherto unrecognized role in preventing the establishment of a latent-like state

Prediction of social structure and genetic relatedness in colonies of the facultative polygynous stingless bee Melipona bicolor (Hymenoptera, Apidae)

Stingless bee colonies typically consist of one single-mated mother queen and her worker offspring. The stingless bee Melipona bicolor (Hymenoptera: Apidae) shows facultative polygyny, which makes this species particularly suitable for testing theoretical expectations concerning social behavior. In this study, we investigated the social structure and genetic relatedness among workers from eight natural and six manipulated colonies of M. bicolor over a period of one year. The populations of M. bicolor contained monogynous and polygynous colonies. The estimated genetic relatedness among workers from monogynous and polygynous colonies was 0.75 ± 0.12 and 0.53 ± 0.16 (mean ± SEM), respectively. Although the parental genotypes had significant effects on genetic relatedness in monogynous and polygynous colonies, polygyny markedly decreased the relatedness among nestmate workers. Our findings also demonstrate that polygyny in M. bicolor may arise from the adoption of related or unrelated queens