Observações anatômicas em plantas de Coffea arabica L. obtidas por enraizamento de estacas

Uma forma para se obter diminuição significativa de tempo e recursos despendidos nos programas de melhoramento de Coffea arabica L. é a clonagem de híbridos F1 por meio de estacas caulinares. Alguns estudos, em diferentes instituições, foram realizados buscando-se definir um método eficiente para esse tipo de clonagem. Com o objetivo de verificar-se a presença de barreiras anatômicas ao enraizamento de estacas caulinares do cafeeiro e a origem das raízes adventícias, bem como compara-las às raízes provenientes de plantas obtidas por semeadura, foram realizadas análises anatômicas no Departamento de Biologia da Universidade Federal de Lavras (UFLA), Lavras, MG. Utilizaram-se estacas caulinares de cafeeiro dos cultivares Acaiá e Rubi e mudas obtidas por semeadura direta e por estaquia. Os cortes realizados nas estacas caulinares mostraram não existirem barreiras anatômicas ao enraizamento adventício. Nas estacas enraizadas, a origem do primórdio radicular foi próxima aos tecidos vasculares. Cortes histológicos nas raízes formadas nas estacas e nas raízes de mudas obtidas por semeadura confirmaram que elas apresentam as mesmas estruturas primárias.

Genetic divergence of tomato subsamples

Understanding the genetic variability of a species is crucial for the progress of a genetic breeding program and requires characterization and evaluation of germplasm. This study aimed to characterize and evaluate 101 tomato subsamples of the Salad group (fresh market) and two commercial controls, one of the Salad group (cv. Fanny) and another of the Santa Cruz group (cv. Santa Clara). Four experiments were conducted in a randomized block design with three replications and five plants per plot. The joint analysis of variance was performed and characteristics with significant complex interaction between control and experiment were excluded. Subsequently, the multicollinearity diagnostic test was carried out and characteristics that contributed to severe multicollinearity were excluded. The relative importance of each characteristics for genetic divergence was calculated by the Singh's method (Singh, 1981), and the less important ones were excluded according to Garcia (1998). Results showed large genetic divergence among the subsamples for morphological, agronomic and organoleptic characteristics, indicating potential for genetic improvement. The characteristics total soluble solids, mean number of good fruits per plant, endocarp thickness, mean mass of marketable fruit per plant, total acidity, mean number of unmarketable fruit per plant, internode diameter, internode length, main stem thickness and leaf width contributed little to the genetic divergence between the subsamples and may be excluded in future studies.

Recent advances in pharmacologic study of natural anticancer agents in China

In this paper a number of anticancer agents of natural origin will be presented. Hydroxycamtothecin (HCPT) was found to produce a strong inhibitory action on a variety of animal tumors. It is also effective for treatment of patients with gastric carcinoma, liver carcinoma, tumor of head and neck or leukemia. Pharmacologic studies showed that it could depress S phase of tumor cells significantly and cause formation of cellular chromatid breaks. By means of alkaline elution and nick translation methods it has been proved that HCPT induced DNA singlo strand breaks remarkably. Homoharringyonine (hhrt) was shown to be effective against acute leukemia. Recent experiments in tumor-bearing mice inidcated that (HHRT) could diminish tumor metastasis. Using molecular hybridization technique it was demonstrated that (HHRT) decreased the content of c-myc RNA in the cytoplasm but not in the nuclei. Lycobetaine (LBT) poddrddrf dytnh inhibitory effects on a number of ascites tumors. In clinical trials it was against ovarian and gastric carcinomas. It is able to intercalate into DNA. Oxalysine (OXL) is a new antibiotic and shown to be effective against tumor metastatis. When used in combination with 5-FU, its anticancer action could be enhanced. Other natural compounds such as indirubin, ß-elemene, irisquinone, oridonine, norcantharidin and PSP have been also found to possess antitumor action.

Trichomonas vaginalis and Tritrichomonas foetus: interaction with fibroblasts and muscle cells - new insights into parasite-mediated host cell cytotoxicity

Trichomonas vaginalis and Tritrichomonas foetus are parasitic, flagellated protists that inhabit the urogenital tract of humans and bovines, respectively. T. vaginalis causes the most prevalent non-viral sexually transmitted disease worldwide and has been associated with an increased risk for human immunodeficiency virus-1 infection in humans. Infections by T. foetus cause significant losses to the beef industry worldwide due to infertility and spontaneous abortion in cows. Several studies have shown a close association between trichomonads and the epithelium of the urogenital tract. However, little is known concerning the interaction of trichomonads with cells from deeper tissues, such as fibroblasts and muscle cells. Published parasite-host cell interaction studies have reported contradictory results regarding the ability of T. foetus and T. vaginalis to interact with and damage cells of different tissues. In this study, parasite-host cell interactions were examined by culturing primary human fibroblasts obtained from abdominal biopsies performed during plastic surgeries with trichomonads. In addition, mouse 3T3 fibroblasts, primary chick embryo myogenic cells and L6 muscle cells were also used as models of target cells. The parasite-host cell cultures were processed for scanning and transmission electron microscopy and were tested for cell viability and cell death. JC-1 staining, which measures mitochondrial membrane potential, was used to determine whether the parasites induced target cell damage. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling staining was used as an indicator of chromatin damage. The colorimetric crystal violet assay was performed to ana-lyse the cytotoxicity induced by the parasite. The results showed that T. foetus and T. vaginalis adhered to and were cytotoxic to both fibroblasts and muscle cells, indicating that trichomonas infection of the connective and muscle tissues is likely to occur; such infections could cause serious risks to the infected host.

Influence of the Paracoccidioides brasiliensis14-3-3 and gp43 proteins on the induction of apoptosis in A549 epithelial cells

The fungal strain Paracoccidioides brasiliensisremains viable inside of epithelial cells and can induce apoptosis in this population. However, until now, the molecules that participate in this process remained unknown. Thus, this study evaluated the contribution of two P. brasiliensismolecules, the 14-3-3 and glycoprotein of 43 kDa proteins, which had been previously described as extracellular matrix adhesins and apoptosis inductors in human pneumocytes. Accordingly, epithelial cells were treated with these molecules for different periods of time and the expression of the apoptosis regulating-proteins Bak, Bax, Bcl-2, p53 and caspases were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labelling, flow cytometry and real-time polymerase chain reaction analysis. Our results demonstrated that treatment with these molecules induces apoptosis signalling in pulmonary epithelial cells, showing the same pattern of programmed cell-death as that observed during infection with P. brasiliensis. Thus, we could conclude that P. brasiliensisuses these molecules as virulence factors that participate not only in the fungal adhesion process to host cells, but also in other important cellular mechanisms such as apoptosis.

Funções e ordem da razão dos nutrientes no estabelecimento de normas DRIS em laranjeira 'Valência'

O sistema integrado de diagnose e recomendação DRIS (Diagnosis and Recommendation Integrated System) tem sido uma boa alternativa para a avaliação do estado nutricional de diversas culturas. O objetivo deste trabalho foi calcular índices DRIS mediante o uso de três métodos (Beaufils, Elwali & Gascho e Jones) de cálculo das funções das razões dos nutrientes e de dois critérios (Letzch e Nick) para a escolha da ordem da razão dos nutrientes, em pomares comerciais irrigados de laranjeira 'Valência'. Informações sobre a produtividade, porta-enxerto e teor de N, P, K, Ca, Mg, S, Fe, Mn, Cu, Zn e B em folhas de ramos não-frutíferos de cada talhão foram processadas nos anos de 1994 a 1998. O índice de balanço nutricional (IBN) foi calculado pela média da soma dos valores em módulo dos índices nutricionais e do índice de matéria seca. Os índices DRIS e IBN foram calculados e os valores de r² (produtividade em função de IBN) foram obtidos para cada método. O método Jones para cálculo do DRIS apresentou melhor correlação com a produtividade, e o critério Nick mostrou-se mais eficiente na escolha da ordem da razão dos nutrientes. Populações específicas, com pequeno número de observações, padronizadas quanto ao porta-enxerto e referentes a um ou dois anos de amostragem foliar e produção foram bancos de dados eficientes para a obtenção das normas DRIS.

DRIS norms for 'Valencia' sweet orange on three rootstocks

Diagnosis and Recommendation Integrated System (DRIS) applies nutrient ratios instead of the isolated concentration values of each nutrient in interpretation of tissue analysis. The objectives of this research were to establish adequate DRIS norms for 'Valencia' sweet orange irrigated commercial groves budded on three rootstocks and correlate indexes of nutrition balance with yield. Experiments were conducted in São Paulo State, Brazil. Rootstocks Rangpur lime, Caipira sweet orange, and Poncirus trifoliata, with more than six years old and yield above 40 ton ha-1 were utilized. Data referred to yield, tree spacing, rootstock and foliar concentrations of N, P, K, Ca, Mg, S, Fe, Mn, Cu, Zn, and B in non fruiting terminals for each grove were processed for the years 1994 through 1998. DRIS indexes were calculated by Nick criterion for choosing the ratio order of the nutrients and Jones calculation method of the ratio functions. Indexes of nutritional balance calculated from DRIS norms presented high correlation with yield for the three scion/rootstock combinations. DRIS norms defined in this research are valid, since leaf sampling is done on non fruiting terminals and the grove is irrigated.

Caracterização morfoagronômica e coeficientes de trilha de caracteres componentes da produção em mandioca

Os objetivos deste trabalho foram caracterizar agronomicamente cem clones de mandioca (Manihot esculenta Crantz) e calcular os coeficientes de trilha entre a produção de raízes tuberosas e cinco componentes da produção, de modo a auxiliar na seleção de clones superiores. O experimento foi conduzido em área experimental da Universidade Federal de Lavras, em 2005 e 2006. Cem clones de mandioca foram avaliados em delineamento látice quadrado 10x10. A unidade experimental foi constituída por quatro plantas espaçadas de 1,0x0,6 m. As análises estatísticas foram realizadas considerando-se as oito características individualmente, utilizando-se o teste de Scott-Knott para agrupamento das médias. Os clones 87 e 88 mostraram-se promissores tanto para serem utilizados em cruzamentos quanto para fixação como novas cultivares em virtude do excelente desempenho nas características comprimento, número e produção de raízes tuberosas por planta. A análise de trilha mostrou que o número de raízes por planta e o peso total da parte aérea podem ser utilizados como critérios na seleção indireta para produção de raízes tuberosas em mandioca.

Identificação dos genes Ty-2 e Ty-3 de resistência a begomovírusem genótipos de tomateiro

O objetivo deste trabalho foi identificar a presença dos genes Ty-2 e Ty-3, de resistência a begomovírus, em acessos de tomateiro do Banco de Germoplasma de Hortaliças da Universidade Federal de Viçosa. Os oligonucleotídeos TO302 F/R e FLUW25 F/R foram utilizados em reações de PCR, para verificar a presença de marcadores relacionados aos genes Ty-2 e Ty-3, respectivamente. Observou-se a presença do gene Ty-2, em heterozigose na subamostra BGH-6881 (Solanum peruvianum), e do gene Ty-3, em homozigose nas subamostras BGH-6878, BGH-6897 (S. lycopersicum) e em heterozigose na subamostra BGH-6881. A identificação dos genes de resistência, com reações de PCR, representa um avanço para os programas de melhoramento de tomateiro no Brasil.

Seleção simultânea para qualidade do fruto e resistência à requeima em progênies de tomateiro

O objetivo deste trabalho foi avaliar critérios de seleção em progênies de cruzamento entre a cultivar de tomateiro Santa Clara (Solanum lycopersicum) e a espécie silvestre S. habrochaites f. glabratum, quanto a atributos de qualidade dos frutos e de resistência à requeima (Phytophthora infestans). As famílias foram avaliadas em delineamento de blocos ao acaso, em dois ensaios, com duas repetições e seis testemunhas comuns a ambos os ensaios. Ganhos diretos e indiretos foram estimados entre famílias F2:3 para seleção simultânea quanto à resistência à requeima, determinada pela quantificação da área abaixo da curva de progresso da doença (AACPD), e quanto à acidez titulável e aos teores de sólidos solúveis dos frutos. Os critérios de seleção proporcionaram ganhos genéticos satisfatórios, adequados ao ideótipo proposto de decréscimo na AACPD e de incremento nos valores médios de sólidos solúveis e acidez titulável. A seleção direta e indireta e o índice de Mulamba & Mock resultam em ganhos individuais mais equilibrados e em maiores ganhos totais.

Seleção de acessos de tomateiro resistentes à pinta-preta pela análise de agrupamento das curvas de progresso da doença

O objetivo deste trabalho foi selecionar acessos resistentes à pinta-preta (Alternaria tomatophila) por meio da análise de agrupamento das curvas de progresso da doença em tomateiro (Solanum lycopersicum). Foram avaliados 134 acessos de tomateiro do Banco de Germoplasma de Hortaliças da Universidade Federal de Viçosa (BGH-UFV), no delineamento de blocos ao acaso, além das testemunhas suscetíveis 'Débora' e 'Santa Clara'. As plantas foram inoculadas com uma mistura de conídios de diferentes isolados de Alternaria spp. e avaliadas regularmente quanto à severidade da doença a cada três dias após a inoculação, no total de seis avaliações. Ajustou-se o modelo logístico aos dados de severidade da pinta-preta, e as estimativas obtidas para a incidência final da doença (B1) e a taxa de progresso da doença (B3) foram submetidas à análise de variância multivariada (Manova). As médias dessas estimativas, para cada acesso, foram submetidas à análise de agrupamento. Foram formados 24 grupos distintos com base no agrupamento das curvas de progresso da doença, o que possibilitou identificar os acessos BGH-2143, BGH-2235, BGH-2270 e BGH-2118 de tomateiro como potenciais fontes de resistência à pinta-preta.

Potencial de hibridação entre acessos de tomateiro para pré-melhoramento quanto à resistência à requeima

Resumo: O objetivo deste trabalho foi avaliar genitores com potencial de hibridação para o pré-melhoramento do tomateiro (Solanum lycopersicum) quanto à resistência à requeima. Foram utilizados seis acessos de tomateiro (BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 e BGH-2343) como genitores resistentes e 15 híbridos F1 originários destes genitores. Utilizou-se o delineamento de blocos ao acaso, com três repetições. As plantas foram inoculadas com uma mistura de esporângios de Phytophthora infestans, agente etiológico da requeima, na concentração de 5x103 esporângios mL-1. A área abaixo da curva de progresso da doença foi utilizada para avaliar a resistência. Realizou-se a análise dialélica, tendo-se considerado o efeito de genótipos como fixo. Estimou-se a capacidade geral e específica de combinação dos acessos. O padrão de resistência dos genitores e da maioria dos F1 foi o mesmo que o das testemunhas resistentes. Foram observados: variabilidade genética aditiva entre os genitores, predominância de efeitos gênicos não aditivos e desvios de dominância bidirecional no controle do caráter. A frequência de alelos favoráveis e divergentes para resistência à requeima é maior nos acessos BGH-2117, BGH-2127 e BGH-2343.

Apoptosis of sensory neurons and satellite cells after sciatic nerve transection in C57BL/6J mice

The rate of axonal regeneration, after sciatic nerve lesion in adult C57BL/6J mice, is reduced when compared to other isogenic strains. It was observed that such low regeneration was not due just to a delay, since neuronal death was observed. Two general mechanisms of cell death, apoptosis and necrosis, may be involved. By using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) technique, we demonstrated that a large number of sensory neurons, as well as satellite cells found in the dorsal root ganglia, were intensely labeled, thus indicating that apoptotic mechanisms were involved in the death process. Although almost no labeled neurons or satellite cells were observed one week after transection, a more than ten-fold increase in TUNEL labeling was detected after two weeks. The results obtained with the C57BL/6J strain were compared with those of the A/J strain, which has a much higher peripheral nerve regeneration potential. In A/J mice, almost no labeling of sensory neurons or satellite cells was observed after one or two weeks, indicating the absence of neuronal loss. Our data confirm previous observations that approximately 40% of C57BL/6J sensory neurons die after sciatic nerve transection, and indicate that apoptotic events are involved. Also, our observations reinforce the hypothesis that the low rate of axonal regeneration occurring in C57BL/6J mice may be the result of a mismatch in the timing of the neurons need for neurotrophic substances, and production of the latter by non-neuronal cells in the distal stump.

Insulin impairs the maturation of chondrocytes in vitro

The precise nature of hormones and growth factors directly responsible for cartilage maturation is still largely unclear. Since longitudinal bone growth occurs through endochondral bone formation, excess or deficiency of most hormones and growth factors strongly influences final adult height. The structure and composition of the cartilaginous extracellular matrix have a critical role in regulating the behavior of growth plate chondrocytes. Therefore, the maintenance of the three-dimensional cell-matrix interaction is necessary to study the influence of individual signaling molecules on chondrogenesis, cartilage maturation and calcification. To investigate the effects of insulin on both proliferation and induction of hypertrophy in chondrocytes in vitro we used high-density micromass cultures of chick embryonic limb mesenchymal cells. Culture medium was supplemented with 1% FCS + 60 ng/ml (0.01 µM) insulin and cultures were harvested at regular time points for later analysis. Proliferating cell nuclear antigen immunoreactivity was widely detected in insulin-treated cultures and persisted until day 21 and [³H]-thymidine uptake was highest on day 14. While apoptosis increased in control cultures as a function of culture time, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-labeled cells were markedly reduced in the presence of insulin. Type II collagen production, alkaline phosphatase activity and cell size were also lower in insulin-treated cultures. Our results indicate that under the influence of 60 ng/ml insulin, chick chondrocytes maintain their proliferative potential but do not become hypertrophic, suggesting that insulin can affect the regulation of chondrocyte maturation and hypertrophy, possibly through an antiapoptotic effect.

Retinal protective effects of topically administered agmatine on ischemic ocular injury caused by transient occlusion of the ophthalmic artery

Agmatine, an endogenous polyamine and putative neuromodulator, is known to have neuroprotective effects on various neurons in the central nervous system. We determined whether or not topically administered agmatine could reduce ischemic retinal injury. Transient ocular ischemia was achieved by intraluminal occlusion of the middle cerebral artery of ddY mice (30-35 g) for 2 h, which is known to also induce occlusion of the ophthalmic artery. In the agmatine group (N = 6), a 1.0 mM agmatine-containing ophthalmic solution was administered four times daily for 2 weeks before occlusion. In the control group (N = 6), a 0.1% hyaluronic acid ophthalmic solution was instilled at the same times. At 22 h after reperfusion, the eyeballs were enucleated and the retinal sections were stained by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Transient ocular ischemia induced apoptosis of retinal cells in the entire retinal layer, and topically administered agmatine can significantly reduce this ischemic retinal injury. The proportion of apoptotic cells was definitely decreased (P < 0.001; Kruskal-Wallis test). Overall, we determined that topical agmatine application effectively decreases retinal damage in an in vivo ocular ischemic injury model. This implies that agmatine is a good candidate as a direct neuroprotective agent for eyes with ocular ischemic diseases.