Peduncle and fruit yield, in six cropping seasons, of early dwarf cashew tree clones irrigated with different water regimes

The objective of this work was to evaluate peduncle and fruit yield in clone MS 076 and in a clonal population of drip-irrigated, early dwarf cashew trees propagated by layering, in six cropping seasons. In order to meet the increased water requirements of the crop resulting from plant growth and development, irrigation during the dry season was performed daily according to the following water regime: 15 min/plant/day during the 1st year, 30 min/plant/day during the 2nd year, 45 min/plant/day during the 3rd year and 60 min/plant/day during all subsequent years. Water was supplied by one drip emitter/plant, at an (adjustable) flow rate of 36 L/h.The research was carried out in Fortaleza-Ceará, Brazil, and a random block design was utilized, with five replicates and split-plots. The clones were assigned to plots and the cropping seasons were considered as subplots. The clonal population was superior to the clone only with regard to number of nut shells (NNS), and solely in the first season. The clone was superior to the population as to NNS and peduncle yield (PY) in the second season, and also with regard to the three evaluated traits - NNS, PY, and nut shell yield, in the last three cropping seasons.

Microenxertia em cultivares de manga

Com a finalidade de se testar a viabilidade do método de microenxertia para produzir mudas de mangueira livres do fungo Fusarium subglutinans, agente causal da malformação, foram realizados experimentos utilizando-se do ápice meristemático da cultivar Tommy Atkins. Retirou-se o ápice meristemático do porta-enxerto e colocou-se o ápice meristemático da cultivar-copa, denominando-se essa metodologia de "microenxertia por substituição de ápice meristemático", na qual foram utilizadas as cultivares Coquinho, Espada, Ouro e Ubá como porta-enxertos. O material de propagação utilizado foi retirado de uma planta-matriz da cultivar Tommy Atkins sem sintomas de malformação. Primeiramente, a parte apical dos ramos foi cortada com aproximadamente 3 cm de comprimento. Os meristemas foram colocados em uma solução antioxidante composta de ácido ascórbico, ácido cítrico e L-cisteína, para evitar a oxidação dos compostos fenólicos existentes na manga. Os meristemas apicais foram cortados com comprimento de 2 mm. Em seguida, efetuou-se o corte do meristema apical e de folhas do porta-enxerto, colocando-se o meristema apical sobre o corte do porta-enxerto, recobrindo-se com Parafilm®. Demonstrou-se com a técnica de microenxertia a possibilidade de formação de plantas-matrizes, para implantação de jardim clonal em condições de viveiro protegido.

Rigitano: nova cultivar de umezeiro para porta-enxerto de pessegueiro

A pesquisa brasileira não havia desenvolvido, até o presente momento, um porta-enxerto clonal para a cultura do pessegueiro com características agronômicas desejáveis, especialmente com relação à resistência a nematóides-de-galha, facilidade de propagação por estacas herbáceas e indução à melhoria da qualidade dos frutos da cultivar-copa. O presente trabalho tem por objetivo apresentar a cultivar Rigitano de umezeiro, selecionada e aprovada para constituir um novo porta-enxerto para a cultura do pessegueiro. Identificada inicialmente como 'Clone 10', a cultivar Rigitano é resultante de um amplo projeto de pesquisa, realizado a partir de 1998 em colaboração com material procedente do Instituto Agronômico de Campinas (IAC), na Faculdade de Ciências Agrárias e Veterinárias (FCAV/UNESP), Câmpus de Jaboticabal (SP). Os trabalhos de seleção e multiplicação para validação técnico-científica final iniciaram-se com experimentos de propagação por estacas herbáceas, cujos resultados indicaram viabilidade do método nas quatro estações do ano, nas condições climáticas de Jaboticabal (SP). A enxertia com o pessegueiro 'Aurora-1', borbulhia em escudo ou escudo modificado, demonstrou ser viável em porta-enxertos de maior diâmetro (± 10 mm). Em condições de campo, 'Rigitano' revelou-se o menos vigoroso dos clones de umezeiro testados. Além disso, 'Rigitano' é resistente a Meloidogyne javanica e M. incognita, entretanto é suscetível a Mesocriconema xenoplax. Os resultados de campo, como porta-enxerto da cv. Aurora-1 de pessegueiro, revelam boa produtividade e frutos com boas qualidades pomológicas e tecnológicas. Os resultados de pesquisa obtidos revelam amplas possibilidades de sucesso da cv. Rigitano em sua validação como novo porta-enxerto de pessegueiro, bem como seu uso visando à redução do espaçamento de plantio e à produção de frutos de melhor qualidade.

A single epidermal stem cell strategy for safe ex vivo gene therapy.

There is a widespread agreement from patient and professional organisations alike that the safety of stem cell therapeutics is of paramount importance, particularly for ex vivo autologous gene therapy. Yet current technology makes it difficult to thoroughly evaluate the behaviour of genetically corrected stem cells before they are transplanted. To address this, we have developed a strategy that permits transplantation of a clonal population of genetically corrected autologous stem cells that meet stringent selection criteria and the principle of precaution. As a proof of concept, we have stably transduced epidermal stem cells (holoclones) obtained from a patient suffering from recessive dystrophic epidermolysis bullosa. Holoclones were infected with self-inactivating retroviruses bearing a COL7A1 cDNA and cloned before the progeny of individual stem cells were characterised using a number of criteria. Clonal analysis revealed a great deal of heterogeneity among transduced stem cells in their capacity to produce functional type VII collagen (COLVII). Selected transduced stem cells transplanted onto immunodeficient mice regenerated a non-blistering epidermis for months and produced a functional COLVII. Safety was assessed by determining the sites of proviral integration, rearrangements and hit genes and by whole-genome sequencing. The progeny of the selected stem cells also had a diploid karyotype, was not tumorigenic and did not disseminate after long-term transplantation onto immunodeficient mice. In conclusion, a clonal strategy is a powerful and efficient means of by-passing the heterogeneity of a transduced stem cell population. It guarantees a safe and homogenous medicinal product, fulfilling the principle of precaution and the requirements of regulatory affairs. Furthermore, a clonal strategy makes it possible to envision exciting gene-editing technologies like zinc finger nucleases, TALENs and homologous recombination for next-generation gene therapy.

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

Background: Coxiella burnetii is a highly clonal microorganism which is difficult to culture, requiring BSL3 conditions for its propagation. This leads to a scarce availability of isolates worldwide. On the other hand, published methods of characterization have delineated up to 8 different genomic groups and 36 genotypes. However, all these methodologies, with the exception of one that exhibited limited discriminatory power (3 genotypes), rely on performing between 10 and 20 PCR amplifications or sequencing long fragments of DNA, which make their direct application to clinical samples impracticable and leads to a scarce accessibility of data on the circulation of C. burnetii genotypes. Results: To assess the variability of this organism in Spain, we have developed a novel method that consists of a multiplex (8 targets) PCR and hybridization with specific probes that reproduce the previous classification of this organism into 8 genomic groups, and up to 16 genotypes. It allows for a direct haracterization from clinical and environmental samples in a single run, which will help in the study of the different genotypes circulating in wild and domestic cycles as well as from sporadic human cases and outbreaks. The method has been validated with reference isolates. A high variability of C. burnetii has been found in Spain among 90 samples tested, detecting 10 different genotypes, being those adaA negative associated with acute Q fever cases presenting as fever of intermediate duration with liver involvement and with chronic cases. Genotypes infecting humans are also found in sheep, goats, rats, wild boar and ticks, and the only genotype found in cattle has never been found among our clinical samples. Conclusions: This newly developed methodology has permitted to demonstrate that C. burnetii is highly variable in Spain. With the data presented here, cattle seem not to participate in the transmission of C. burnetii to humans in the samples studied, while sheep, goats, wild boar, rats and ticks share genotypes with the human population.

Avaliação genética de indivíduos e progênies de cupuaçuzeiro no estado do Pará e estimativas de parâmetros genéticos

O presente trabalho relata a avaliação genotípica de progênies de cupuaçuzeiro, no Estado do Pará, para os caracteres número de frutos (NF) em quatro safras, intensidade de ocorrência de vassoura- de-bruxa na inflorescência (VBI) e nos frutos (VBF), e peso de vassoura-de-bruxa (PVB). Apresenta também estimativas de parâmetros genéticos que permitem inferir sobre o controle genético e nível de variabilidade genética presente no material avaliado. Todos os caracteres apresentaram considerável variabilidade genética, com coeficientes de variação genética variando de 27% a 88% no âmbito de progênie e de 38% a 123% no âmbito individual. Isto revela excelentes possibilidades para a seleção nessa população experimental híbrida. As estimativas de herdabilidade individual no sentido restrito, em uma safra, variaram de 25% a 54%, e as repetibilidades individuais para NF equivaleram a 35%. Com as quatro safras realizadas, a herdabilidade em nível individual aumentou para 48%, propiciando acurácia seletiva de 70%, para a seleção de indivíduos. O ganho em eficiência, quando se usa mais de cinco safras, é praticamente desprezível. Para NF, ganhos acima de 60% podem ser obtidos com a seleção dos cinco melhores indivíduos. Poderão ser selecionados indivíduos com produção anual de 17 frutos, valor muito superior à média geral de 10 frutos, encontrada nos plantios comerciais. Verificam-se ganhos genéticos bastante superiores quando se faz a propagação clonal dos melhores indivíduos em relação ao que se verifica quando se realiza a propagação sexuada. Para o melhor indivíduo, o ganho genético aumenta de 75.5% para 88.3%, ou seja, de 17 para quase 19 frutos por planta. Isto revela um grande potencial para a clonagem comercial de cupuaçuzeiro. Para os caracteres VBI e VBF, verificaram-se altas herdabilidades individuais no sentido restrito com valores variando entre 30% e 54%. Isto revela o excelente potencial da seleção recorrente para melhorar, gradativamente, o nível de resistência. Parece suficiente considerar na seleção apenas o número de vassouras, não sendo necessário considerar o peso. A correlação entre resistência no fruto e na inflorescência foi alta (0.84), indicando algum controle genético comum aos dois caracteres. Foram identificadas progênies superiores, simultaneamente, para produção de frutos e resistência à vassoura.

Diversidade genética de aceroleiras (Malphigia emarginata D.C.), utilizando marcadores moleculares RAPD e características morfoagronômicas

O conhecimento da variabilidade genética e da relação entre diferentes acessos de aceroleira é importante para maximizar o uso dos recursos genéticos para futuros programas de melhoramento. O objetivo deste trabalho foi avaliar a divergência genética entre 48 acessos de aceroleira, por meio de marcadores moleculares RAPD e características morfoagronômicas. Foram utilizados 25 iniciadores, possibilitando obter 108 marcadores, sendo observadas 92 marcas polimórficas. Os marcadores obtidos foram analisados, usando os métodos de otimização de Tocher e hierárquico UPGMA, que gerou um dendrograma utilizando o índice de Jaccard. Os resultados mostraram uma concordância parcial entre os métodos de agrupamentos estudados, com a formação de 14 grupos. Os acessos ACE 023 e ACE 033 foram os mais distintos, apresentando distância genética de 0,58. A análise comparativa dos agrupamentos revelou que os marcadores RAPD, associados com características morfoagronômicas, foram eficientes para a discriminação dos acessos e que houve uma variabilidade genética potencial para o programa de melhoramento genético e informações úteis, como a indicação de acessos promissores para avaliação clonal.

Miniestaquia seriada na propagação da goiabeira 'Paluma'

A miniestaquia seriada é uma técnica empregada na propagação comercial do Eucalyptus, e sua utilização tem mostrado efeito positivo sobre o enraizamento em clones de baixo potencial de enraizamento. O objetivo deste trabalho foi avaliar a técnica da miniestaquia seriada na sobrevivência e no enraizamento de miniestacas, e no vigor das mudas de goiabeira 'Paluma'. Foram conduzidos dois experimentos sob delineamento de blocos casualizados, com quatro repetições e cinco plantas por parcela. Aos sessenta e dois dias após o estaqueamento, verificou-se que as miniestacas tiveram sobrevivência acima de 90% e 90% de enraizamento. As mudas produzidas por miniestaquia de subcultivo (mudas obtidas a partir de minicepas formadas de miniestacas enraizadas), aos 138 dias após o estaqueamento, tinham altura inferior à das mudas provenientes de miniestacas de primeiro cultivo (mudas obtidas a partir de minicepas formadas pelo processo de estaquia convencional). Por outro lado, as avaliações realizadas aos 145 dias após o estaqueamento indicaram que as mudas produzidas por miniestaquia de subcultivo apresentaram o mesmo vigor de crescimento em área foliar, número de folhas e massa de matéria seca da parte aérea e do sistema radicular em relação às mudas produzidas por miniestacas de primeiro cultivo.

Allelic diversity and population structure in Vibrio cholerae O139 Bengal based on nucleotide sequence analysis

Comparative analysis of gene fragments of six housekeeping loci, distributed around the two chromosomes of Vibrio cholerae, has been carried out for a collection of 29 V. cholerae O139 Bengal strains isolated from India during the first epidemic period (1992 to 1993). A toxigenic O1 ElTor strain from the seventh pandemic and an environmental non-O1/non-O139 strain were also included in this study. All loci studied were polymorphic, with a small number of polymorphic sites in the sequenced fragments. The genetic diversity determined for our O139 population is concordant with a previous multilocus enzyme electrophoresis study in which we analyzed the same V. cholerae O139 strains. In both studies we have found a higher genetic diversity than reported previously in other molecular studies. The results of the present work showed that O139 strains clustered in several lineages of the dendrogram generated from the matrix of allelic mismatches between the different genotypes, a finding which does not support the hypothesis previously reported that the O139 serogroup is a unique clone. The statistical analysis performed in the V. cholerae O139 isolates suggested a clonal population structure. Moreover, the application of the Sawyer's test and split decomposition to detect intragenic recombination in the sequenced gene fragments did not indicate the existence of recombination in our O139 population.

Dendritic Cells Cause Bone Lesions in a New Mouse Model of Histiocytosis.

Langerhans cell histiocytosis (LCH) is a rare disease caused by the clonal accumulation of dendritic Langerhans cells, which is often accompanied by osteolytic lesions. It has been reported that osteoclast-like cells play a major role in the pathogenic bone destruction seen in patients with LCH and these cells are postulated to originate from the fusion of DCs. However, due to the lack of reliable animal models the pathogenesis of LCH is still poorly understood. In this study, we have established a mouse model of histiocytosis- recapitulating human disease for osteolytic lesions seen in LCH patients. At 12 weeks after birth, severe bone lesions were observed in our multisystem histiocytosis (Mushi) model, when CD8α conventional dendritic cells (DCs) are transformed (MuTuDC) and accumulate. Most importantly, our study demonstrates that bone loss in LCH can be accounted for the transdifferentiation of MuTuDCs into functional osteoclasts both in vivo and in vitro. Moreover, we have shown that injected MuTuDCs reverse the osteopetrotic phenotype of oc/oc mice in vivo. In conclusion, our results support a crucial role of DCs in bone lesions in histiocytosis patients. Furthermore, our new model of LCH based on adoptive transfer of MuTuDC lines, leading to bone lesions within 1-2 weeks, will be an important tool for investigating the pathophysiology of this disease and ultimately for evaluating the potential of anti-resorptive drugs for the treatment of bone lesions.

GROWTH AND NUTRITION OF CACAO SEEDLINGS INFLUENCED BY ZINC APLICATION IN SOIL

ABSTRACT Levels of Zn in tropical soils profoundly influences growth and nutrition of tree crops. Research was undertaken to assess the effect of soil Zn on growth and nutrition of clonal cacao tree seedlings of PH 16. Three acidic Oxisol soils differing in texture were used with nine doses of Zn (0, 1, 2, 4, 8, 16, 32, 48, and 64 mg dm-3). Rooted clonal seedlings were grown in plastic pot with 11 dm-3 of the soils at varying Zn levels for 240 days. At harvest growth (dry matter mass of leaves, stems, shoots, roots, and total) and nutrient concentrations were determined. The clonal cacao seedlings showed differences for production of dry matter mass and foliar nutrient concentrations for P, K, Ca, Mg, Mn, Fe, Zn, and Cu. There was Zn toxicity in all soils.

Marked increase in incidence for bloodstream infections due to Escherichia coli, a side effect of previous antibiotic therapy in the elderly.

We conducted a survey including 3334 bloodstream infections (BSIs) due to E. coli diagnosed in 2005-2014 at a stable cohort of hospitals. Marked increases in incidence were observed for community-acquired (CA) BSIs in patients aged >75 years, CA-BSIs of digestive origin in patients aged 60-74 years, healthcare-associated BSIs, and BSIs associated with ESBL (extended-spectrum B-lactamase)-producing E. coli (ESBLEc). Using MLST, we studied the genetic diversity of 412 BSI isolates recovered during the 2014 survey: 7 major sequence type complexes (STCs) were revealed in phylogenetic group B2, 3 in group A/B1 and 2 in group D. Among the 31 ESBLEc isolates, 1/3 belonged to STC 131. We searched for possible associations between clonal groups, clinical determinants and characteristics of BSIs: isolates from groups B2 (except STC 131) and D were susceptible to antibiotics and associated with BSIs of urinary origin in patients <60 years. STC 131 and group A/B1 isolates were multi-drug resistant and associated with CA-BSIs of digestive origin in patients aged 60-74 with a recent history of antibiotic treatment. STC 131 isolates were associated with HCA-BSIs in patients with recent/present hospitalization in a long-stay unit. We provide a unique population-based picture of the epidemiology of E. coli BSI. The aging nature of the population led to an increase in the number of cases caused by the B2 and D isolates generally implicated in BSIs. In addition, the association of a trend toward increasing rates of gut colonization with multi drug-resistant isolates revealed by the rise in the incidence of BSIs of digestive origin caused by STC 131 and A/B1 (STCs 10, 23, and 155) isolates, and a significant increase in the frequency of BSIs in elderly patients with recent antibiotic treatment suggested that antibiotic use may have contributed to the growing incidence of BSI.

Cerebriforms cells in cerebrum: an unusual finding

Background. Mycosis Fungoides (MF) is the most common cutaneous T-cell lymphoma, and large cell trasformation (tMF) is an adverse prognostic event. Extra-cutaneous dissemination can occur in the course of the disease, but dissemination to the central nervous system (CNS) is uncommon. Moreover, CNS lymphomas are overall rare and most often of B-cell phenotype. We report a case of CNS large T-cell lymphoma presenting as multiple cerebral lesions in a patient with a history of MF. Methods. We report a case of a 33-year-old woman, known since the age of 16 for erythematous plaques thought to be atopic dermatitis, who developed, end 2012, multiple nodular skin lesions and peripheral adenopathies. Two skin lesions were biopsied simultaneously, and diagnosed as MF and tMF. A lymph node biopsy showed dermatopathic changes without lymphoma (Stage IIB). She received local treatment (UVB, PUVA and radiation therapy) and interferon therapy, and experienced almost complete remission. In 2015 neurological symptoms lead to evidence multiple cerebral lesions, suspicious for lymphoma, evaluated by stereotaxic biopsies. We compared histopathological and molecular features of these with previous skin specimens. After negative bone marrow staging biopsy, she was recently started on chemotherapy (MATRIX). Short follow-up shows rapidly worsening clinical conditions. Results. One of the initial skin biopsies showed atypical lymphoid cells with epidermotropism, Pautrier abcesses and CD4+ CD30- phenotype; the other revealed diffuse dermal infiltration by predominantly large cerebriform tumor cells with high proliferative fraction, and CD2−CD3 −CD4+/−CD7−CD30+ALK- EMA- non-cytotoxic immunophenotype. Altogether, these results led us to diagnose MF and tMF, respectively. The brain was infiltrated by large atypical lymphoid cells with cerebriform nuclei, somewhat anaplastic features and perivascular distribution. By immunohistochemistry, tumor cells were highly proliferative, with a CD2−CD3+CD5−CD7+CD30+ activated cytotoxic immunophenotype. A diagnosis of CD30+ cytotoxic peripheral T-cell lymphoma was retained. TRG and TRB clonality analyses revealed clonal rearrangements in skin and CNS biopsies, with identical patterns in both skin specimens but only minimally overlapping profiles when compared to the CNS sample. Der Pathologe 6 ? 2015 | 633 Conclusions. The reported case illustrates an uncommon finding of a CNS T-cell lymphoma in a patient with previous MF, questioning the clonal relationship between the two diseases and challenging the adequate classification of this CNS lymphoma as either a progression or a de novo lymphoma. Despite differences in immunophenotype and clonality patterns, this CNS lymphoma could possibly represent an aggressive divergent evolution of a primary cutaneous T-cell lymphoma. Additional sequencing is ongoing to try to solve the question.

Tangential migration of neuronal precursors of glutamatergic neurons in the adult mammalian brain.

In a classic model of mammalian brain formation, precursors of principal glutamatergic neurons migrate radially along radial glia fibers whereas GABAergic interneuron precursors migrate tangentially. These migration modes have significant implications for brain function. Here we used clonal lineage tracing of active radial glia-like neural stem cells in the adult mouse dentate gyrus and made the surprising discovery that proliferating neuronal precursors of glutamatergic granule neurons exhibit significant tangential migration along blood vessels, followed by limited radial migration. Genetic birthdating and morphological and molecular analyses pinpointed the neuroblast stage as the main developmental window when tangential migration occurs. We also developed a partial "whole-mount" dentate gyrus preparation and observed a dense plexus of capillaries, with which only neuroblasts, among the entire population of progenitors, are directly associated. Together, these results provide insight into neuronal migration in the adult mammalian nervous system.

The Stealthy Superbug: the Role of Asymptomatic Enteric Carriage in Maintaining a Long-Term Hospital Outbreak of ST228 Methicillin-Resistant Staphylococcus aureus.

UNLABELLED: Whole-genome sequencing (WGS) of 228 isolates was used to elucidate the origin and dynamics of a long-term outbreak of methicillin-resistant Staphylococcus aureus (MRSA) sequence type 228 (ST228) SCCmec I that involved 1,600 patients in a tertiary care hospital between 2008 and 2012. Combining of the sequence data with detailed metadata on patient admission and movement confirmed that the outbreak was due to the transmission of a single clonal variant of ST228, rather than repeated introductions of this clone into the hospital. We note that this clone is significantly more frequently recovered from groin and rectal swabs than other clones (P < 0.0001) and is also significantly more transmissible between roommates (P < 0.01). Unrecognized MRSA carriers, together with movements of patients within the hospital, also seem to have played a major role. These atypical colonization and transmission dynamics can help explain how the outbreak was maintained over the long term. This "stealthy" asymptomatic colonization of the gut, combined with heightened transmissibility (potentially reflecting a role for environmental reservoirs), means the dynamics of this outbreak share some properties with enteric pathogens such as vancomycin-resistant enterococci or Clostridium difficile. IMPORTANCE: Using whole-genome sequencing, we showed that a large and prolonged outbreak of methicillin-resistant Staphylococcus aureus was due to the clonal spread of a specific strain with genetic elements adapted to the hospital environment. Unrecognized MRSA carriers, the movement of patients within the hospital, and the low detection with clinical specimens were also factors that played a role in this occurrence. The atypical colonization of the gut means the dynamics of this outbreak may share some properties with enteric pathogens.