Photoinduced Photosensitizer-Antibody Conjugates Kill HIV Env-Expressing Cells, Also Inactivating HIV.

HIV-infected cells persist for decades in patients administered with antiretroviral therapy (ART). Meanwhile, an alarming surge in drug-resistant HIV viruses has been occurring. Addressing these issues, we propose the application of photoimmunotherapy (PIT) against not only HIV Env-expressing cells but also HIV. Previously, we showed that a human anti-gp41 antibody (7B2) conjugated to cationic or anionic photosensitizers (PSs) could specifically target and kill the HIV Env-expressing cells. Here, our photolysis studies revealed that the binding of photoimmunoconjugates (PICs) on the membrane of HIV Env-expressing cells is sufficient to induce necrotic cell death due to physical damage to the membrane by singlet oxygen, which is independent of the type of PSs. This finding persuaded us to study the virus photoinactivation of PICs using two HIV-1 strains, X4 HIV-1 NL4-3 and JR-CSF virus. We observed that the PICs could destroy the viral strains, probably via physical damage on the HIV envelope. In conclusion, we report the application of PIT as a possible dual-tool for HIV immunotherapy and ART by killing HIV-expressing cells and cell-free HIV, respectively

Phenotypic variability among pumpkin accessions in the Brazilian semiarid

The aim of this study was to estimate the phenotypic divergence among fourteen pumpkins accessions of the cucurbits Germplasm Active Bank of Embrapa Semiárido, in order to support breeding program to these species. Nine morphoagronomic descriptors were analyzed by the principal components and by UPGMA techniques. In the analysis of principal components, the formation of three contrasting clusters was observed, each one composed of two accessions. There occurred also the formation of one core group composed of five accessions, being the accessions called 683 and 684 collected in Bahia and the other in Maranhão. The most divergent accessions were 620, 624 and 748. The variables of the greatest contribution to the genetic divergence were diameter, weight and average length of the fruit. In the UPGMA analysis, obtained from the symmetric matrices generated by the Mahalanobis distance for the nine descriptors, there was the formation of one core group, composed of ten accessions, whose similarity was 45.71%, with geographically divergent accessions presenting the greatest similarity. Three accessions were not clustered with others. The study identified phenotypic variability for the analyzed characters and the absence of relationship between place of collection and phenotypic diversity

The nuclear envelope protein, LAP1B, is a novel protein phosphatase 1 substrate

Protein phosphatase 1 (PP1) binding proteins are quintessential regulators, determining substrate specificity and defining subcellular localization and activity of the latter. Here, we describe a novel PP1 binding protein, the nuclear membrane protein lamina associated polypeptide 1B (LAP1B), which interacts with the DYT1 dystonia protein torsinA. The PP1 binding domain in LAP1B was here identified as the REVRF motif at amino acids 55-59. The LAP1B:PP1 complex can be immunoprecipitated from cells in culture and rat cortex and the complex was further validated by yeast co-transformations and blot overlay assays. PP1, which is enriched in the nucleus, binds to the N-terminal nuclear domain of LAP1B, as shown by immunocolocalization and domain specific binding studies. PP1 dephosphorylates LAP1B, confirming the physiological relevance of this interaction. These findings place PP1 at a key position to participate in the pathogenesis of DYT1 dystonia and related nuclear envelope-based diseases.publishe

Detection and Quantification of Leptospira interrogans in Hamster and Rat Kidney Samples: Immunofluorescent Imprints versus Real-time PCR

A major limitation in the clinical management and experimental research of leptospirosis is the poor performance of the available methods for the direct detection of leptospires. In this study, we compared real-time PCR (qPCR), targeting the lipL32 gene, with the immunofluorescent imprint method (IM) for the detection and quantification of leptospires in kidney samples from the rat and hamster experimental models of leptospirosis. Using a virulent strain of Leptospira interrogans serovar Copenhageni, a chronic infection was established in the rat model, which were euthanized 28 days post-infection, while the hamster model simulated an acute infection and the hamsters were euthanized eight days after inoculation. Leptospires in the kidney samples were detected using culture isolation, qPCR and the IM, and quantified using qPCR and the IM. In both the acute and chronic infection models, the correlation between quantification by qPCR and the IM was found to be positive and statistically significant (P<0.05). Therefore, this study demonstrates that the IM is a viable alternative for not only the detection but also the quantification of leptospires, particularly when the use of qPCR is not feasible

An intriguing shift occurs in the novel protein phosphatase 1 binding partner, TCTEX1D4: evidence of positive selection in a pika model

T-complex testis expressed protein 1 domain containing 4 (TCTEX1D4) contains the canonical phosphoprotein phosphatase 1 (PPP1) binding motif, composed by the amino acid sequence RVSF. We identified and validated the binding of TCTEX1D4 to PPP1 and demonstrated that indeed this protein is a novel PPP1 interacting protein. Analyses of twenty-one mammalian species available in public databases and seven Lagomorpha sequences obtained in this work showed that the PPP1 binding motif 90RVSF93 is present in all of them and is flanked by a palindromic sequence, PLGS, except in three species of pikas (Ochotona princeps, O. dauurica and O. pusilla). Furthermore, for the Ochotona species an extra glycosylation site, motif 96NLS98, and the loss of the palindromic sequence were observed. Comparison with other lagomorphs suggests that this event happened before the Ochotona radiation. The dN/dS for the sequence region comprising the PPP1 binding motif and the flanking palindrome highly supports the hypothesis that for Ochotona species this region has been evolving under positive selection. In addition, mutational screening shows that the ability of pikas TCTEX1D4 to bind to PPP1 is maintained, although the PPP1 binding motif is disrupted, and the N- and C-terminal surrounding residues are also abrogated. These observations suggest pika as an ideal model to study novel PPP1 complexes regulatory mechanisms.publishe

Ganhos genéticos para caracteres de raiz em populações de cenoura nos sistemas orgânico e convencional de produção

O objetivo deste trabalho foi verificar os ganhos genéticos para caracteres de raiz em populações de cenoura cultivadas nos sistemas orgânico e convencional. Os experimentos foram conduzidos na Embrapa Hortaliças, Brasília (DF). Duas populações de cenoura derivadas da cultivar Brasília e de origem comum até 2002, foram subdivididas em duas populações, avaliadas e selecionadas, por oito gerações consecutivas, nos verões de 2000 a 2007. Em 2008, amostras de sementes das populações, provenientes de cada ciclo de seleção, foram semeadas em campo e conduzidas sob manejo orgânico e convencional de produção, em delineamento de blocos casualizados com nove tratamentos, quatro repetições e parcelas de 1 m². Aos 90 dias após a semeadura, 20 raízes por parcela foram colhidas para a avaliação dos caracteres comprimento, diâmetro do xilema e do floema; comprimento da extensão do ombro verde; massa fresca; presença de halo; formato de ponta e de ombro e coloração do xilema e floema. Foi realizada análise de variância com determinação da interação entre tratamentos e sistemas de produção, agrupamento de médias entre os tratamentos, e calculados os ganhos reais com a seleção. Pôde-se verificar que nos oito anos de seleção não houve ganhos significativos para os caracteres estudados nas duas populações. Com isso, conclui-se que os caracteres avaliados já se encontram fixados nas duas populações estudadas. Verificou-se que a seleção não precisa ser realizada nos dois sistemas de cultivo, orgânico e convencional, possibilitando diminuição de recursos financeiros e de mão-de-obra empregados no melhoramento.The objective of this work was to evaluate the genetic gain with the selection of root characters of carrot populations cultivated in organic and conventional production systems. The experiments were carried out at Embrapa Hortaliças, Brasília, Brazil. Two carrot populations derived from Brasília cultivar and with common origin until 2002, were separate in two populations and evaluated for eight generations during 2000 to 2007. In 2008, seed samples of the population in each cycle of selection were sowed in the field in both organic and conventional production systems, in a randomized blocks design with four replications of nine treatments and plots of 1 m². After 90 days of sowing, 20 roots per plot were harvested for the evaluation of the length, xylem and phloem diameter, green shoulder length, fresh mass, presence of halo, tip and shoulder format, and the color parameters of xylem and phloem. Variance analysis was carried out to determine the interaction between treatments and production systems, grouping of means among treatments, and the real gain with the selection was estimated. In the last eight years of selection, a significant gain was not observed on the studied characters in the two populations. So we concluded that those traits are already quite developed and stabilized in both populations. The selection doesn't need to be accomplished in both areas of organic and conventional cultivation, making possible the decrease of financial and labor resources utilized in the breeding

Measurement of the inclusive and dijet cross-sections of b-jets in pp collisions at sqrt(s) = 7 TeV with the ATLAS detector

The inclusive and dijet production cross-sections have been measured for jets containing b-hadrons (b-jets) in proton-proton collisions at a centre-of-mass energy of sqrt(s) = 7 TeV, using the ATLAS detector at the LHC. The measurements use data corresponding to an integrated luminosity of 34 pb^-1. The b-jets are identified using either a lifetime-based method, where secondary decay vertices of b-hadrons in jets are reconstructed using information from the tracking detectors, or a muon-based method where the presence of a muon is used to identify semileptonic decays of b-hadrons inside jets. The inclusive b-jet cross-section is measured as a function of transverse momentum in the range 20 < pT < 400 GeV and rapidity in the range |y| < 2.1. The bbbar-dijet cross-section is measured as a function of the dijet invariant mass in the range 110 < m_jj < 760 GeV, the azimuthal angle difference between the two jets and the angular variable chi in two dijet mass regions. The results are compared with next-to-leading-order QCD predictions. Good agreement is observed between the measured cross-sections and the predictions obtained using POWHEG + Pythia. MC@NLO + Herwig shows good agreement with the measured bbbar-dijet cross-section. However, it does not reproduce the measured inclusive cross-section well, particularly for central b-jets with large transverse momenta.Comment: 10 pages plus author list (21 pages total), 8 figures, 1 table, final version published in European Physical Journal

HIV/AIDS-associated visceral leishmaniasis in patients from an endemic area in Central-west Brazil

An increase in morbidity associated with visceral leishmaniasis (VL) in human immunodeficiency virus (HIV)/AIDS patients has been described in Africa and the Mediterranean. Despite the high endemicity of VL and HIV-1/AIDS in Brazil, this association has not been thoroughly investigated. Our aim was to evaluate the epidemiologic and clinical characteristics of VL-HIV-1/AIDS cases from Central-west [Mato Grosso do Sul (MS)] Brazil. Medical records of 23 VL-HIV-1/AIDS patients were reviewed. Patients were predominantly adult males (87%) and 34.8% of the patients were intravenous drug users (IVDU). Leishmaniasis was the first opportunistic infection in 60% of the HIV-1 patients. Fever occurred in all patients, although splenomegaly and hepatomegaly were absent in 21.7% of the cases. CD4+ T-cell counts were below 200 cells/mm³ in 80% of the cases and the counts did not increase after clinical remission despite antiretroviral therapy. The first drug chosen to treat the cases was antimonial, but the therapeutic regimen was altered to amphotericin B in 12 of 17 cases due to side effects. Relapses were reported in 56.5% of the patients. IVDU may constitute an important risk factor for the transmission of both diseases in MS. VL-HIV-1/AIDS patients in MS share similar clinical characteristics as those from other endemic regions worldwide. Thus, these findings are critical for improving the surveillance of VL-HIV/AIDS patients