Phenotypic Diversity and Genetic Parameters of Coffea canephora Clones.

The simultaneous analysis of the maximum number of chemical elements present in plant tissues provides more comprehensive information about their chemical constitution and increases the number of characteristics for the selection process in various plant breeding programs. The objective of this study was to analyze productivity, grain yield, and concentration of chemical elements in tissues of Coffea canephora clones to study phenotypic diversity and estimate genetic parameters for use in breeding. This experiment was carried out in Manaus, Amazonas, Brazil, in randomized blocks with four replications. The concentrations of elements in various organs were quantified using total reflection X-ray fluorescence (TXRF). Genetic parameters and genetic divergence were estimated, and genotypes were clustered using the UPGMA hierarchical method and non-metric multidimensional scaling analysis. The study allowed us to differentiate the performance of the clones in terms of the absorption of essential and non-essential chemical elements for plant development and to analyze the correlation of the characteristics in the selection process. TXRF efficiently characterizes the presence and concentration of multiple elements, aiding genotype discrimination for C. canephora improvement.

Acidente vascular encefálico isquêmico devido à síndrome do anticorpo antifosfolípide em paciente lúpica - um relato de caso / schemic stroke due to antiphospholipid syndrome in a lupus patient - a case report

RESUMO: Lúpus eritematoso sistêmico é uma doença de origem autoimune que apresenta um amplo espectro de manifestações clínicas causadas pelo estado inflamatório crônico. A associação com a síndrome do anticorpo antifosfolípide ocorre em aproximadamente 36% dos pacientes lúpicos. Nesses pacientes, a doença de base e o uso crônico de corticosteroides contribuem para aterosclerose acelerada, uma complicação que agrega risco cardiovascular. Relatamos um caso de acidente vascular encefálico em paciente de 25 anos portadora de lúpus eritematoso sistêmico e síndrome do anticorpo antifosfolípide. (AU)

ABSTRACT: Systemic lupus erythematosus is an autoimmune disease that presents a wide spectrum of clinical manifestations caused by the chronic inflammatory state. Association with antiphospholipid antibody syndrome occurs in approximately 36% of lupus patients. In these patients, the underlying disease and the chronic use of corticosteroids contribute to accelerated atherosclerosis, a complication that adds increased cardiovascular risk. We report a case of stroke in a 25-year-old patient with systemic lupus erythematosus and antiphospholipid antibody syndrome. (AU)

Leishmania braziliensis isolated from disseminated leishmaniasis patients downmodulate neutrophil function.

AIMS: The polymorphism observed in Leishmania braziliensis is associated with different clinical forms of leishmaniasis. Neutrophils (PMNs) participate in the pathogenesis of leishmania infection, and here, we evaluate neutrophil function after infection with isolates of L. braziliensis from cutaneous leishmaniasis (CL) or disseminated leishmaniasis (DL) patients. METHODS AND RESULTS: Neutrophils from 30 healthy subjects (HS) were infected with isolates of L. (V.) braziliensis obtained from three CL and three DL patients. They were infected at the ratio of 3:1 parasites per neutrophil, and leishmania uptake was evaluated by microscopy. The neutrophil activation markers and oxidative burst by expression of dihidrorhodamine (DHR) were evaluated by flow cytometry and cytokine production by ELISA. The frequency of infected cells and the number of amastigotes were higher in neutrophils infected with CL isolates compared to DL isolates (P < 0.05). The DHR and CD66b expression after infection with DL isolate was lower than with CL isolates. There was no difference regarding chemokine production. CONCLUSION: The L. (V.) braziliensis isolates of DL induced lower respiratory burst and neutrophils activation markers compared with CL isolates which may contribute to parasite survival and dissemination in DL patients.

Genetic diversity among accessions of sacha inchi (Plukenetia volubilis) by phenotypic characteristics analysis / Diversidade genética entre acessos de sacha inchi (Plukenetia volubilis) por análise de características fenotípicas

Sacha inchi (Plukenetia volubilis) is native to the Amazon region and has a high seed content of mono and polyunsaturated fatty acids, making it interesting for the pharmaceutical and cosmetic industry. The purpose of this study was to analyze sacha inchi genetic diversity and describe accessions based on phenotypic characteristics. Fruits and seeds of 25 accessions from the sacha inchi genebank of Embrapa Amazônia Ocidental in Manaus, Amazonas state, were sampled and biometrically measured. The data were subjected to analysis of variance, Mahalanobis distance, canonical correlation, and genetic diversity among and within accessions by analysis of molecular variance (AMOVA). There were significant differences among the means of the analyzed traits, but no significant canonical correlation for the groups of traits. According to AMOVA, approximately 60% of the observed variation was within accessions. The results showed variability among accessions, and that the variation within accessions should be explored to obtain best results in breeding programs.

Sacha inchi (Plukenetia volubilis) é nativa da região amazônica e suas sementes tem um alto teor de ácidos graxos mono e poliinsaturados, tornando-a interessante para a indústria farmacêutica e cosmética. O objetivo deste estudo foi analisar a diversidade genética de sacha inchi e caracterizar os acessos com base em características fenotípicas. Foi realizada coleta e biometria de frutos e sementes de 25 acessos do banco de germoplasma de sacha inchi da Embrapa Amazônia Ocidental em Manaus-AM. Os dados foram submetidos a análise de variância, distância de Mahalanobis, correlação canônica e diversidade genética por análise de variância molecular (AMOVA). Houve diferenças significativas entre as médias das variáveis analisadas, contudo, não houve correlação canônica significativa para os grupos de variáveis. De acordo com AMOVA, aproximadamente 60% da variação observada esteve dentro de acessos. Os resultados mostram variabilidade entre acessos, sendo importante explorar a variação intra-acessos para obter melhores resultados em programas de melhoramento.

Retinol and Betacarotene Status in Mother-Infant Dyads and Associations between Them.

BACKGROUND/AIMS: Assessing the diet and biochemical indicators of vitamin A deficiency (VAD) in high-risk populations is crucial in cases where this deficiency is mainly caused by chronically inadequate intake. This study aimed to determine the retinol and betacarotene status in mother-infant dyads, and to evaluate the associations between them. METHODS: Umbilical cord serum, maternal serum, and colostrum were collected from 134 healthy mothers living in a risk region for VAD. Vitamin A and betacarotene were quantified by liquid chromatography, and dietary information was collected using a food frequency questionnaire. RESULTS: Although the overall mean intakes of vitamin A and betacarotene were considered adequate, 16% of the women had insufficient intake. Mean retinol levels were also adequate, yet low levels were diagnosed in about 8% of the mothers, based on maternal serum and colostrum, and in 16% of the cord serum samples. Retinol and betacarotene were positively associated in cord serum (p = 0.004), maternal serum (p = 0.041), and colostrum (p < 0.001) but was not associated with dietary intake. CONCLUSIONS: A diagnosis of adequacy based on mean biochemical and dietary data of this population in fact masks the marginal vitamin A status presented by mothers and children.

Seed-Specific Stable Expression of the α-AI1 Inhibitor in Coffee Grains and the In Vivo Implications for the Development of the Coffee Berry Borer.

Genetic transformation of coffee (Coffea spp.), the second most traded commodity worldwide, is an alternative approach to introducing features that cannot be introgressed by traditional crossings. The transgenic stability, heritability and quantitative and spatial expression patterns of the seed-specific promoter phytohemagglutinin (PHA-L) from Phaseolus vulgaris were characterized in genetically modified C. arabica expressing the α-amylase inhibitor-1 (α-AI1) gene. The α-AI1 inhibitor shows considerable activity toward digestive enzymes of the coffee berry borer (CBB) Hypothenemus hampei. This insect pest expends its life cycle almost entirely in coffee berries. Transgene containment in the fruit is important to meeting food and environmental safety requirements for releasing genetically modified (GM) crops. PCR analysis of T2 coffee plants showed a Mendelian single-copy segregation pattern. Ectopic transgene expression was only detected in coffee grains, as demonstrated by reverse transcription-PCR analysis of different plant tissues. An intense immunocytochemical signal associated with α-AI1 protein expression was localized to endospermic cells. In addition, a delay in the larval development of CBB was observed after challenging transgenic coffee seeds with the insect. These results indicate that the PHA-L promoter might be a useful tool in coffee for the seed-specific expression of genes related to coffee bean productivity, quality and pest protection. The biotechnological applicability of the α-AI1 gene for controlling CBB is also discussed. This work is the first report showing a seed-specific transgene expression in coffee plants.

Employing in vitro directed molecular evolution for the selection of α-amylase variant inhibitors with activity toward cotton boll weevil enzyme.

Numerous species of insect pests attack cotton plants, out of which the cotton boll weevil (Anthonomus grandis) is the main insect in Brazil and must be controlled to avert large economic losses. Like other insect pests, A. grandis secretes a high level of α-amylases in the midgut lumen, which are required for digestion of carbohydrates. Thus, α-amylase inhibitors (α-AIs) represent a powerful tool to apply in the control of insect pests. Here, we applied DNA shuffling and phage display techniques and obtained a combinatorial library containing 108 α-AI variant forms. From this library, variants were selected exhibiting in vitro affinity for cotton boll weevil α-amylases. Twenty-six variant sequences were cloned into plant expression vectors and expressed in Arabidopsis thaliana. Transformed plant extracts were assayed in vitro to select specific and potent α-amylase inhibitors against boll weevil amylases. While the wild type inhibitors, used to create the shuffled library, did not inhibit the A. grandis α-amylases, three α-AI mutants, named α-AIC3, α-AIA11 and α-AIG4 revealed high inhibitory activities against A. grandis α-amylases in an in vitro assay. In summary, data reported here shown the potential biotechnology of new α-AI variant genes for cotton boll weevil control.

Macrophages participate in host protection and the disease pathology associated with Leishmania braziliensis infection.

BACKGROUND: Leishmania preferentially infects macrophages, which allow the parasite to multiply but can also kill the parasite. Although the T cell response in human leishmaniasis is well-characterized, little is known about the concomitant macrophage behavior. The aim of this study was to characterize the macrophage immune response after Leishmania braziliensis infection in cells derived from cutaneous leishmaniasis (CL) or mucosal leishmaniasis (ML) patients, subclinical individuals (SC) and healthy control subjects (HS). METHODS: Peripheral blood mononuclear cell-derived macrophages from the different groups were exposed to L. braziliensis in vitro and were evaluated for susceptibility to Leishmania infection, ability to kill Leishmania and chemokine/cytokine production. Nitric Oxide (NO) and superoxide (O2-) levels in the supernatant of infected macrophage cultures were monitored. RESULTS: After exposure to L. braziliensis, peripheral blood mononuclear cell-derived macrophages from SC individuals showed a lower infection rate and a smaller number of intracellular amastigotes compared to cells from CL and ML patients. Macrophages from CL and ML patients produced more chemokines and TNF-α than those from the SC group. Production of NO and O2- were detected but did not vary significantly among the different groups. CONCLUSIONS: Our data indicate that macrophages play a pivotal role in controlling L. braziliensis infection and in leishmaniasis pathology by secreting pro-inflammatory chemokines/cytokines that activate and recruit T cells, overwhelming the inflammatory response.

Evaluation of the microbicidal activity and cytokines/chemokines profile released by neutrophils from HTLV-1-infected individuals.

Human T cell lymphotropic virus type-1 (HTLV-1) induces activation and spontaneous proliferation of T cells with production of type-1 pro-inflammatory cytokines. It modifies the immune response to other antigens and increases susceptibility to infectious diseases. However, little is known about innate immunity in HTLV-1 infection. HTLV-1-infected individuals have higher spontaneous neutrophil activation than HTLV-1-seronegative individuals, as shown by the nitroblue tetrazolium (NBT) assay. This study was conducted to evaluate neutrophil function in HTLV-1-infected individuals. Participants in the study included 18 HTLV-1-infected individuals and 14 HTLV-1-seronegative controls. We evaluated the ability of neutrophils (PMNs) to control a parasite infection, to produce peroxynitrite, cytokines and chemokines and to express activation markers in cultures when stimulated with LPS or infected with Leishmania. When compared with the control group, there was no difference in the percentage of PMNs infected with Leishmania or in the number of amastigotes/100 PMNs in HTLV-1-infected individuals. The microbicidal activity of the PMNs and the levels of CXCL8 and CCL4 released by these cells did not show a difference between HTLV-1-infected individuals and the control group. In both the HTLV-1 group and the control group, infection with Leishmania or stimulation of PMNs led to cellular activation. These observations suggest that neutrophils from HTLV-1-infected individuals have preserved their ability to become activated and to produce chemokines and peroxynitrite after stimulation and that the susceptibility to infection by intracellular Leishmania amazonensis in HTLV-1-infected individuals does not depend on impairment of neutrophil function.