Stable expression of shRNA for the control of recombinant adenovirus replication.

Preventing the replication of adenovirus could have practical uses, such as controlling infection with wild-type virus or in applications involving recombinant vectors. Mainly transient methods have been used to inhibit adenovirus replication, including siRNA or drugs. Here, we tested whether stable expression of shRNA designed to target hexon, Iva2, or pol can inhibit the replication of a recombinant adenoviral vector, Ad-LacZ (serotype 5, E1/E3 deleted), in 293T cells. Significant knockdown correlating with reduced Ad-LacZ replication was achieved only when hexon was targeted. Cell sorting and isolation of cellular clones further accentuated knockdown of the hexon transcript, reduced protein levels by more than 90%, and diminished adenovirus production. As visualized by transmission electron microscopy, the cellular clone expressing the hexon-specific shRNA yielded 89.2% fewer particles compared to the parental 293T cells. Full scale production followed by purification revealed a 90.2% reduction in Ad-LacZ biological titer. These results support the notion that stable expression of shRNA can be used as a means to control adenovirus replication.

Biocontrol potential of actinobacteria against Pantoea ananatis, the causal agent of maize white spot disease.

Pantoea ananatis is the causal agent of maize white spot, a foliar disease responsible for significant maize yield reduction worldwide, especially in Brazil. In general, the maize foliar diseases control involves the adoption of resistant genotypes and pesticides application. However, the use of agrochemicals can significantly cause increase production costs, damage to human health and negative environmental impacts. In this sense, the use of biological control agents has been considered among the most promising eco-friendly technologies for sustainable agriculture. Actinobacteria, particularly of Streptomyces genus, has been widely recognized as agroindustrially important microorganism due to its potential in producing diverse range of secondary metabolites, including antibiotics and enzymes. Thus, the aim of this work is to characterize and to evaluate the potential of soil actinobacteria for P. ananatis control. We observed that 59 actinobacteria strains (85%) exhibited proteolytic or chitinolytic activity. Only the strains Streptomyces pseudovenezuelae ACSL 470, that also exhibited high proteolytic activity, S. novaecaesareae ACSL 432 and S. laculatispora ACP 35 demonstrated high or moderate antagonist activity in vitro against P. ananatis. Temporal analysis of metabolites produced by these strains growth in different liquid media indicated greater antibacterial activity at 72 h. In this condition, chromatographic and mass spectrometry analysis revealed that S. pseudovenezuelae ACSL 470 strain produced neomycin, an aminoglycoside antibiotic that displayed high bactericidal activity in vitro against P. ananatis. This is the first report of actinobacteria acting as potential microbial antagonists for P. ananatis control. Further studies are needed to determine the control efficacy of maize white spot disease by Streptomyces strains or their metabolites in greenhouse and field conditions.

Public policy interventions for preventing and treating obesity: scoping review.

CONTEXT: Obesity is a multicausal social problem and a pandemic, and it presents a public health challenge in many countries. Hence, public health interventions have been used in an endeavour to prevent and/or control increased obesity among populations. OBJECTIVE: This review study aimed to provide an overview of the academic literature and to analyze the strategies involved in the main public policies focused on preventing and controlling obesity in a number of countries. DATA SOURCES: MEDLINE/PubMed, Web of Science, and LILACS databases were searched. DATA EXTRACTION: Original studies were included for which the core objective was related to real-life public policy interventions for obesity. DATA ANALYSIS: The studies were organized according to their characteristics, and the qualitative analysis was based on the categorization proposed by the author Poulain. The review included 41 studies and identified 15 types of interventions focused on obesity; the largest proportion of actions were developed in school and city environments and were geared toward behavioral change, supported by direct or indirect government action. It was observed that many strategies focused on environmental line through regulatory and legislative measures, and health promotion geared toward the individual was based on the dissemination of information as a means for behavioral change. A smaller focus was given to individual care and treatment, and participative actions in the community. CONCLUSION: The design of public health strategies applicable to obesity as proposed by Poulain provides a valid model for evaluating interventions. The behavioral approach involving guidance through health education toward a healthy lifestyle prevails in public policies, indicating a movement toward accountability of individuals. However, it is necessary to deepen the debate on the social structures that determine obesity (and which limit possibilities of choice), aligned with cultural change regarding its occurrence, and to employ care strategies based on scientific evidence and which focus on the needs of the subjects.

Biocontrol potential of actinobacteria against Pantoea ananatis, the causal agent of maize white spot disease / Potencial de biocontrole de actinobactérias contra Pantoea ananatis, agente causal da doença mancha branca do milho

Pantoea ananatis is the causal agent of maize white spot, a foliar disease responsible for significant maize yield reduction worldwide, especially in Brazil. In general, the maize foliar diseases control involves the adoption of resistant genotypes and pesticides application. However, the use of agrochemicals can significantly cause increase production costs, damage to human health and negative environmental impacts. In this sense, the use of biological control agents has been considered among the most promising eco-friendly technologies for sustainable agriculture. Actinobacteria, particularly of Streptomyces genus, has been widely recognized as agroindustrially important microorganism due to its potential in producing diverse range of secondary metabolites, including antibiotics and enzymes. Thus, the aim of this work is to characterize and to evaluate the potential of soil actinobacteria for P. ananatis control. We observed that 59 actinobacteria strains (85%) exhibited proteolytic or chitinolytic activity. Only the strains Streptomyces pseudovenezuelae ACSL 470, that also exhibited high proteolytic activity, S. novaecaesareae ACSL 432 and S. laculatispora ACP 35 demonstrated high or moderate antagonist activity in vitro against P. ananatis. Temporal analysis of metabolites produced by these strains growth in different liquid media indicated greater antibacterial activity at 72 h. In this condition, chromatographic and mass spectrometry analysis revealed that S. pseudovenezuelae ACSL 470 strain produced neomycin, an aminoglycoside antibiotic that displayed high bactericidal activity in vitro against P. ananatis. This is the first report of actinobacteria acting as potential microbial antagonists for P. ananatis control. Further studies are needed to determine the control efficacy of maize white spot disease by Streptomyces strains or their metabolites in greenhouse and field conditions.

Pantoea ananatis é o agente causal da mancha branca do milho, doença foliar responsável pela redução significativa da produtividade do milho em todo o mundo, especialmente no Brasil. Em geral, o controle de doenças foliares do milho envolve a adoção de genótipos resistentes e a aplicação de agrotóxicos. No entanto, o uso de agroquímicos pode causar aumento significativo dos custos de produção, danos à saúde humana e impactos ambientais negativos. Nesse sentido, o uso de agentes de controle biológico tem sido considerado uma das tecnologias ecologicamente corretas mais promissoras para a agricultura sustentável. Actinobactérias, particularmente do gênero Streptomyces, têm sido amplamente reconhecidas como microrganismos de importância agroindustrial devido ao seu potencial de produção de diversos metabólitos secundários, incluindo antibióticos e enzimas. Assim, o objetivo deste trabalho é caracterizar e avaliar o potencial de actinobactérias do solo para o controle de P. ananatis. Observamos que 59 cepas de actinobactérias (85%) apresentaram atividade proteolítica ou quitinolítica. Apenas as cepas Streptomyces pseudovenezuelae ACSL 470, que também exibiu alta atividade proteolítica, S. novaecaesareae ACSL 432 e S. laculatispora ACP 35 demonstraram alta ou moderada atividade antagonista in vitro contra P. ananatis. A análise temporal do crescimento dos metabólitos produzidos por essas cepas em diferentes meios líquidos indicou maior atividade antibacteriana em 72 h. Nesta condição, análises cromatográficas e de espectrometria de massa revelaram que a cepa S. pseudovenezuelae ACSL 470 produziu neomicina, um antibiótico aminoglicosídeo que apresentou alta atividade bactericida in vitro contra P. ananatis. Este é o primeiro relato de actinobactérias atuando como potenciais antagonistas microbianos para o controle de P. ananatis. Mais estudos são necessários para determinar a eficácia do controle da doença da mancha branca do milho por cepas de Streptomyces ou seus metabólitos em condições de casa de vegetação e campo.

Stable expression of shRNA for the control of recombinant adenovirus replication

Preventing the replication of adenovirus could have practical uses, such as controlling infection with wild-type virus or in applications involving recombinant vectors. Mainly transient methods have been used to inhibit adenovirus replication, including siRNA or drugs. Here, we tested whether stable expression of shRNA designed to target hexon, Iva2, or pol can inhibit the replication of a recombinant adenoviral vector, Ad-LacZ (serotype 5, E1/E3 deleted), in 293T cells. Significant knockdown correlating with reduced Ad-LacZ replication was achieved only when hexon was targeted. Cell sorting and isolation of cellular clones further accentuated knockdown of the hexon transcript, reduced protein levels by more than 90%, and diminished adenovirus production. As visualized by transmission electron microscopy, the cellular clone expressing the hexon-specific shRNA yielded 89.2% fewer particles compared to the parental 293T cells. Full scale production followed by purification revealed a 90.2% reduction in Ad-LacZ biological titer. These results support the notion that stable expression of shRNA can be used as a means to control adenovirus replication.

Elevated proportion of TLR2- and TLR4-expressing Th17-like cells and activated memory B cells was associated with clinical activity of cerebral cavernous malformations.

BACKGROUND: Recent evidences have suggested the involvement of toll-like receptor (TLR)-4 in the pathogenesis of cerebral cavernous malformations (CCM). Elevated frequency of TLR+T-cells has been associated with neurological inflammatory disorders. As T-cells and B-cells are found in CCM lesions, the objective of the present study was to evaluate the cytokine profile of T-cells expressing TLR2 and TLR4, as well as B-cell subsets, in asymptomatic (CCMAsympt) and symptomatic (CCMSympt) patients. METHODS: For our study, the cytokine profile from TLR2+ and TLR4+ T-cell and B-cell subsets in CCMAsympt and CCMSympt patients was investigated using flow cytometry and ELISA. T-cells were stimulated in vitro with anti-CD3/anti-CD28 beads or TLR2 (Pam3C) and TLR4 (LPS) ligands. RESULTS: CCMSymptc patients presented a higher frequency of TLR4+(CD4+ and CD8+) T-cells and greater density of TLR4 expression on these cells. With regard to the cytokine profile, the percentage of TLR2+ and TLR4+ Th17 cells was higher in CCMSympt patients. In addition, an elevated proportion of TLR4+ Tc-1 cells, as well as Tc-17 and Th17.1 cells expressing TLR2 and TLR4, was observed in the symptomatic patients. By contrast, the percentage of TLR4+ IL-10+CD4+ T cells was higher in the CCMAsympt group. Both Pam3C and LPS were more able to elevate the frequency of IL-6+CD4+T cells and Th17.1 cells in CCMSympt cell cultures. Furthermore, in comparison with asymptomatic patients, purified T-cells from the CCMSympt group released higher levels of Th17-related cytokines in response to Pam3C and, mainly, LPS, as well as after activation via TCR/CD28. Concerning the B-cell subsets, a higher frequency of memory and memory activated B-cells was observed in CCMSympt patients. CONCLUSIONS: Our findings reveal an increase in circulating Th17/Tc-17 cell subsets expressing functional TLR2 and, mainly, TLR4 molecules, associated with an increase in memory B-cell subsets in CCM patients with clinical activity of the disease.

Major Depressive Disorder Enhances Th2 and Th17 Cytokines in Patients Suffering from Allergic Rhinitis and Asthma.

INTRODUCTION: Major depressive disorder (MDD) can impact the severity of allergic rhinitis (AR) and asthma (AA). Here, we evaluated the cytokine production by T-cells from AR and AA patients with or without MDD. The effect of serotonin on the in vitro T-cell response was also evaluated. METHODS: The cytokines produced by activated T-cells were measured by Luminex and flow cytometry. In some cell cultures, serotonin was added. RESULTS: MDD not only enhanced the production of Th2- and Th17-related cytokines, but also, the levels of interleukin (IL)-5 and IL-17 were directly correlated with the severity of depression and anxiety symptoms. As compared with AR, the levels of IL-17 were higher and the release of IL-10 was lower in activated T-cell cultures from AA patients, mainly those with MDD. In AA/MDD patients, the severity of anxiety symptoms and lung disease was directly correlated with Th17-like and hybrid Th2/Th17 cells, but inversely correlated with IL-10-secreting CD4+ T-cells. Finally, the addition of serotonin reduced the production of Th2- and Th17-related cytokines, but elevated IL-10 secretion in cell cultures from both AR and AA patients. CONCLUSIONS: Our findings suggest that not only the occurrence of MDD but also the severity of anxiety symptoms, may adversely affect the outcome of allergic reactions by favoring the production of cytokines implicated in the pathogenesis of AR and AA, a phenomenon that was attenuated by serotonin.

Batzelladine D and norbatzelladine L purified from marine sponge Monanchora arbuscula induce the reversal of fluconazole.

ATP-Binding Cassette (ABC) transporters are the main class of transmembrane transporters involved in pathogenic fungal resistance against chemotherapeutic agents. Herein we report results which show that batzelladine D (1) and norbatzelladine L (2) reverse the fluconazole resistance phenotype mediated by Pdr5p transporter on Saccharomyces cerevisiae. Both alkaloids were able to chemosensitize the Pdr5p-overexpressing strain by synergistic interaction with fluconazole. Both compounds also showed an inhibitory effect on the catalytic activity and on the intracellular accumulation of rhodamine 6G, and did not show significant in vitro mammalian cells toxicity.

Association Between Vitamin D and Adrenal Parameters with Metabolic and Inflammatory Markers in Polycystic Ovary Syndrome.

Vitamin D deficiency has been related with metabolic alterations in polycystic ovary syndrome (PCOS). As well, hyperactivation of adrenal axis can be programmed early in life and could be related later with PCOS development. Our aim was to establish the relationship between vitamin D and adrenal parameters with metabolic alterations and inflammation markers in PCOS. In 73 patients and 33 controls, 25-hydroxyvitamin D (25-OH-D), total and bioavailable testosterone (TT and bioT), androstenedione (A4), SHBG, cortisol, insulin, and C-reactive protein (hs-CRP) were determined; HOMA and lipid accumulation product (LAP) index were calculated. All parameters were higher in patients than in controls, except for SHBG and 25-OH-D which were lower. Binary regression analysis showed that differences in TT, bioT, A4, insulin and HOMA were independent of body mass index and waist circumference but SHBG, hs-CRP, LAP and 25-OH-D were related to body weight and fat distribution. Binary logistic regression analysis showed that cortisol and 25-OH-D could be associated to PCOS development. Correlations found between LAP and insulin, HOMA and hs-CRP confirm it is a good indicator of metabolic complications. Vitamin D and cortisol association to PCOS development justifies future research to understand the role of vitamin D in PCOS and analyze patient's perinatal history and its possible relationship with hyperactivation of adrenal axis in adult life.

Pregnancy favors the expansion of circulating functional follicular helper T Cells.

Pregnancy favors antibody production, and some evidence has suggested a direct effect of estrogen on B cells. The impact of pregnancy on circulating follicular helper T (TFH) cells, typically identified by the expression of CD45RO and CXCR5, has not been previously investigated. Here, the percentage of TFH cells, co-expressing or not PD-1, ICOS, or CXCR3 markers was significantly higher in pregnant women (PW) as compared with non-pregnant ones (nPW). Furthermore, the percentage of CXCR3+ TFH cells able to produce IL-6, IL-21, and IL-10 was significantly higher in PW than nPW. Interestingly, anti-CMV and anti-HBs antibody titers were significantly higher in the plasma of PW and were directly correlated with IL-21-producing CXCR3+ TFH cells. Finally, peripheral estrogen levels, but not progesterone, were positively related to either PD-1+ CXCR3+ TFH cells or plasma anti-CMV and anti-HBs IgG antibodies. In summary, our data suggests a positive effect of pregnancy on the proportion of CD4+ T cell subset specialized in helping B cells. This phenomenon, which could be related to the high estrogen levels produced during pregnancy, may help to explain why pregnancy favor humoral immunity.

Zika Virus Infects, Activates, and Crosses Brain Microvascular Endothelial Cells, without Barrier Disruption.

Zika virus (ZIKV) has been associated to central nervous system (CNS) harm, and virus was detected in the brain and cerebrospinal fluids of microcephaly and meningoencephalitis cases. However, the mechanism by which the virus reaches the CNS is unclear. Here, we addressed the effects of ZIKV replication in human brain microvascular endothelial cells (HBMECs), as an in vitro model of blood brain barrier (BBB), and evaluated virus extravasation and BBB integrity in an in vivo mouse experimental model. HBMECs were productively infected by African and Brazilian ZIKV strains (ZIKVMR766 and ZIKVPE243), which induce increased production of type I and type III IFN, inflammatory cytokines and chemokines. Infection with ZIKVMR766 promoted earlier cellular death, in comparison to ZIKVPE243, but infection with either strain did not result in enhanced endothelial permeability. Despite the maintenance of endothelial integrity, infectious virus particles crossed the monolayer by endocytosis/exocytosis-dependent replication pathway or by transcytosis. Remarkably, both viruses' strains infected IFNAR deficient mice, with high viral load being detected in the brains, without BBB disruption, which was only detected at later time points after infection. These data suggest that ZIKV infects and activates endothelial cells, and might reach the CNS through basolateral release, transcytosis or transinfection processes. These findings further improve the current knowledge regarding ZIKV dissemination pathways.

Posttherapeutic cure criteria in Chagas' disease: conventional serology followed by supplementary serological, parasitological, and molecular tests.

We performed a critical study of conventional serology, followed by supplementary serological, parasitological, and molecular tests, to assess the response to etiologic treatment of Chagas' disease. A group of 94 Chagas' disease patients treated with benznidazole at least 10 years earlier were evaluated from the laboratory and clinical points of view. When conventional serology (enzyme-linked immunosorbent assay [ELISA], indirect immunofluorescence [IIF], and indirect hemagglutination [IHA]) and classic criteria (consistent results with any two of the three tests) or more rigorous criteria (consistent results from the three tests) were used, 10.6% and 8.5% of patients were considered treated and cured (TC) by classic and rigorous criteria, respectively. Patients were then evaluated using supplementary (recombinant ELISA and Trypanosoma cruzi excreted-secreted antigen blotting [TESA-blot]), parasitological (hemoculture), and molecular (PCR) tests. The results of recombinant ELISA were similar to those with the rigorous criterion (three consistent test results). The TESA-blot group showed a higher percentage (21.3%) of negative results than the groups defined by either cure criterion. Hemoculture and PCR gave negative results for all treated and cured (TC) patients, regardless of the criterion used. Recombinant ELISA and TESA-blot tests showed negative results for 70% and 87.5% of the patients categorized as TC by the classic and three-test criteria, respectively. For patients with discordant conventional serology, the supplementary serological and molecular tests were the decisive factor in determining therapeutic failure. Clinical evaluation showed that 62.5% of TC patients presented with the indeterminate form of the disease. Additionally, treated patients with negative TESA-blot results should be reevaluated later with all methodologies used here to verify whether TESA-blot is a reliable way to determine early parasitological cure of Chagas' disease.

Vulvo-cervico-vaginal manifestations and evaluation of Papanicolaou smears in pemphigus vulgaris and pemphigus foliaceus.

BACKGROUND: Vulvo-cervico-vaginal involvement has rarely been reported in pemphigus vulgaris (PV) and has not been reported in pemphigus foliaceus (PF). OBJECTIVES: We sought to evaluate genital lesions and Papanicolaou (Pap) smears in female patients with PV and PF. METHODS: This prospective study includes all consecutive cases of female patients with PV and PF seen from May 2009 to February 2010. Gynecologic examination was performed and Pap smears were collected for cytologic analysis from each patient. RESULTS: A total of 56 patients were given a diagnosis of pemphigus (41 PV and 15 PF). Genital involvement was observed in 9 patients with PV (22%) and the vulva was the most common genital site of involvement. Of these 9 patients, 8 presented with active skin/mucous lesions. Four of 15 patients with PF had genital lesions and vulva was the exclusive site of involvement. Three of 4 patients with PF and genital involvement also showed active cutaneous lesions. Six of 56 patients (5 PV and 1 PF) presented with atypical squamous cells of undetermined significance in Pap smear analysis. Upon further pathologic review, acantholytic cells were seen, confirming the diagnosis of pemphigus. LIMITATIONS: A small number of PF cases were studied. CONCLUSIONS: Vulvar lesions were the second most frequent site of mucous membrane PV. Herein we report the first case to our knowledge of symptomatic genital lesions in a patient with PF. Moreover, acantholytic cells in Pap smears were found in a patient with PF who was in complete remission off therapy with no clinical genital lesions and no circulating anti-desmoglein-1 and anti-desmoglein-3 autoantibodies. Gynecologic evaluation in patients with pemphigus, including a careful evaluation of Pap smears, should be recommended.

Influência de adubações e manejo de adubo verde nos atributos biológicos de solo cultivado com alface (Lactuca sativa L. ) em sistema de cultivo orgânico / Influence of fertilizer, manure and composting on the biological attributes of soil cultivated with lettuce (Lactuca sativa L. ) in an organic crop system

Devido à carência de informações sobre adubação orgânica para alface e uma avaliação quanto à sua influência nos atributos biológicos do ambiente, este trabalho objetivou estudar o efeito de diferentes sistemas de manejo da mucuna preta e o uso de adubação orgânica sólida sobre a diversidade e biomassa microbiana do solo. Os tratamentos consistiram do manejo da mucuna preta, incorporada ou não ao solo, e aplicação de esterco bovino, composto, adubo orgânico comercial e testemunha sem adubação. Os atributos foram avaliados nas seguintes épocas: momento do manejo da mucuna e aos 15, 55 e 105 dias após esse manejo. O manejo e os tratamentos com adubo orgânico sólido não influenciaram de forma significativa na biomassa microbiana e na população bacteriana do solo. Houve aumento na população de fungo coma incorporação da mucuna preta. A incorporação da mucuna preta e o tratamento com composto foram favoráveis para o incremento de fungos micorrízicos. O tratamento com composto, seguido da incorporação da mucuna preta, e o tratamento testemunha proporcionaram população superior de nematoides saprófitas, enquanto a incorporação da mucuna ao solo aumentou as populações de fitonematoides. (AU)

INFLUENCE OF FERTILIZER, MANURE AND COMPOSTING ON THE BIOLOGICAL ATTRIBUTES OF SOIL CULTIVATED WITH LETTUCE (LACTUCA SATIVA L.) IN AN ORGANIC CROP SYSTEM. Due to the lack of information on the application of commercial organic fertilizers versus manure or composting in the growing of lettuce and their influence on biological attributes of the environment, the present study was carried out to verify the effect of different management systems of velvet beans and the use of manure and solid organic composting on the diversity and microbial biomass of soil. The treatments consisted of the management of velvet beans, incorporated or not into the soil, and the application of bovine manure, composting, commercial organic fertilizer and a control treatment without fertilization. The soil attributes were evaluated at the following times: at the time of the velvet bean management, and at 15, 55 and 105 days after this process. The management and solid organic fertilizer treatments had no significant effect on the microbial biomass and bacterial population. For fungi, the incorporation of velvet bean was more efficient, allied to the use of commercial organic fertilizer. The incorporation of velvet bean and the treatment with composting were favorable for the grown of mycorrhizal fungi. Treatment with composting, followed by the incorporation of velvet beans, and the control treatment provided the higher population of saprophytic nematodes, while the incorporation of velvet bean into the soil increased the population of plant parasitic nematodes. (AU)

Influência de adubações e manejo de adubo verde nos atributos biológicos de solo cultivado com alface (Lactuca sativa L.) em sistema de cultivo orgânico

Due to the lack of information on the application of commercial organic fertilizers versus manure or composting in the growing of lettuce and their influence on biological attributes of the environment, the present study was carried out to verify the effect of different management systems of velvet beans and the use of manure and solid organic composting on the diversity and microbial biomass of soil. The treatments consisted of the management of velvet beans, incorporated or not into the soil, and the application of bovine manure, composting, commercial organic fertilizer and a control treatment without fertilization. The soil attributes were evaluated at the following times: at the time of the velvet bean management, and at 15, 55 and 105 days after this process. The management and solid organic fertilizer treatments had no significant effect on the microbial biomass and bacterial population. For fungi, the incorporation of velvet bean was more efficient, allied to the use of commercial organic fertilizer. The incorporation of velvet bean and the treatment with composting were favorable for the grown of mycorrhizal fungi. Treatment with composting, followed by the incorporation of velvet beans, and the control treatment provided the higher population of saprophytic nematodes, while the incorporation of velvet bean into the soil increased the population of plant parasitic nematodes.

Devido à carência de informações sobre adubação orgânica para alface e uma avaliação quanto à sua influência nos atributos biológicos do ambiente, este trabalho objetivou estudar o efeito de diferentes sistemas de manejo da mucuna preta e o uso de adubação orgânica sólida sobre a diversidade e biomassa microbiana do solo. Os tratamentos consistiram do manejo da mucuna preta, incorporada ou não ao solo, e aplicação de esterco bovino, composto, adubo orgânico comercial e testemunha sem adubação. Os atributos foram avaliados nas seguintes épocas: momento do manejo da mucuna e aos 15, 55 e 105 dias após esse manejo. O manejo e os tratamentos com adubo orgânico sólido não influenciaram de forma significativa na biomassa microbiana e na população bacteriana do solo. Houve aumento na população de fungo coma incorporação da mucuna preta. A incorporação da mucuna preta e o tratamento com composto foram favoráveis para o incremento de fungos micorrízicos. O tratamento com composto, seguido da incorporação da mucuna preta, e o tratamento testemunha proporcionaram população superior de nematoides saprófitas, enquanto a incorporação da mucuna ao solo aumentou as populações de fitonematoides.

Influência de adubações e manejo de adubo verde nos atributos biológicos de solo cultivado com alface (Lactuca sativa L. ) em sistema de cultivo orgânico / Influence of fertilizer, manure and composting on the biological attributes of soil cultivated with lettuce (Lactuca sativa L. ) in an organic crop system

Devido à carência de informações sobre adubação orgânica para alface e uma avaliação quanto à sua influência nos atributos biológicos do ambiente, este trabalho objetivou estudar o efeito de diferentes sistemas de manejo da mucuna preta e o uso de adubação orgânica sólida sobre a diversidade e biomassa microbiana do solo. Os tratamentos consistiram do manejo da mucuna preta, incorporada ou não ao solo, e aplicação de esterco bovino, composto, adubo orgânico comercial e testemunha sem adubação. Os atributos foram avaliados nas seguintes épocas: momento do manejo da mucuna e aos 15, 55 e 105 dias após esse manejo. O manejo e os tratamentos com adubo orgânico sólido não influenciaram de forma significativa na biomassa microbiana e na população bacteriana do solo. Houve aumento na população de fungo coma incorporação da mucuna preta. A incorporação da mucuna preta e o tratamento com composto foram favoráveis para o incremento de fungos micorrízicos. O tratamento com composto, seguido da incorporação da mucuna preta, e o tratamento testemunha proporcionaram população superior de nematoides saprófitas, enquanto a incorporação da mucuna ao solo aumentou as populações de fitonematoides.

INFLUENCE OF FERTILIZER, MANURE AND COMPOSTING ON THE BIOLOGICAL ATTRIBUTES OF SOIL CULTIVATED WITH LETTUCE (LACTUCA SATIVA L.) IN AN ORGANIC CROP SYSTEM. Due to the lack of information on the application of commercial organic fertilizers versus manure or composting in the growing of lettuce and their influence on biological attributes of the environment, the present study was carried out to verify the effect of different management systems of velvet beans and the use of manure and solid organic composting on the diversity and microbial biomass of soil. The treatments consisted of the management of velvet beans, incorporated or not into the soil, and the application of bovine manure, composting, commercial organic fertilizer and a control treatment without fertilization. The soil attributes were evaluated at the following times: at the time of the velvet bean management, and at 15, 55 and 105 days after this process. The management and solid organic fertilizer treatments had no significant effect on the microbial biomass and bacterial population. For fungi, the incorporation of velvet bean was more efficient, allied to the use of commercial organic fertilizer. The incorporation of velvet bean and the treatment with composting were favorable for the grown of mycorrhizal fungi. Treatment with composting, followed by the incorporation of velvet beans, and the control treatment provided the higher population of saprophytic nematodes, while the incorporation of velvet bean into the soil increased the population of plant parasitic nematodes.

Comparison between objective assessment and self-assessment of sexual maturation in children and adolescents / Comparação entre avaliação objetiva e autoavaliação da maturação sexual em crianças e adolescentes

OBJETIVO: Avaliar as correlações entre avaliação objetiva e autoavaliação em ambientes diferentes (ambulatorial e escolar). MÉTODOS: Trezentos e dezenove indivíduos, 178 ambulatoriais (96 meninos e 82 meninas) e 141 (73 meninos e 68 meninas) de escolas públicas (8,3-18,7 anos), com 73 indivíduos (39 meninas e 34 meninos) apresentando índice de massa corporal acima do percentil 85 do Centers for Disease Control and Prevention, de 2000. Todos foram examinados, após consentimento informado, em sequência e individualmente, por dois médicos treinados, e então submetidos a autoavaliação, utilizando fotogramas (Tanner). Dos dados obtidos foram calculados os coeficientes de correlação de kappa entre examinadores e a autoavaliação. Para os testes foi adotado p < 5%. RESULTADOS: Não houve diferença significante entre as correlações obtidas no ambulatório e nas escolas e os dois grupos foram analisados em conjunto. As correlações obtidas entre os examinadores foram significantemente maiores que as da autoavaliação, com kappa (e intervalo de confiança) de 0,75 (0,8-0,69) para mamas/genitália entre examinadores contra 0,27 (0,34-0,20) e 0,29 (0,36-0,22) entre os dois examinadores e a autoavaliação (p < 0,0001). CONCLUSÕES: Na amostra estudada, a autoavaliação do estádio puberal não deve substituir a avaliação objetiva feita por profissionais treinados. Um aperfeiçoamento do método de autoavaliação poderia permitir seu uso em estudos populacionais.

OBJECTIVE: To assess the correlations between objective assessment and self-assessment of sexual maturation in the outpatient and school settings. METHODS: Three hundred and nineteen individuals, 178 (96 boys and 82 girls) from an outpatient clinic and 141 (73 boys and 68 girls) from public schools (8.3-18.7 years), of whom 73 individuals (39 girls an d 34 boys) had a body mass index above the 85th percentile, according to 2000 CDC Growth Chart, were analyzed. All of them were examined sequentially and individually by two trained physicians after a written consent form was signed by parents or surrogates, and then submitted to self-assessment using pictures (Tanner stages). Kappa coefficients between examiners and the self-assessment were calculated based on the collected data. A p value < 5% was established as statistically significant. RESULTS: No significant difference was observed between correlations obtained from the outpatient clinic and schools, and both groups were combined for analysis. The correlations obtained by examiners were significantly higher than those from self-assessment, with a kappa coefficient (and confidence interval) of 0.75 (0.8-0.69) for breasts/genitals across examiners against 0.27 (0.34-0.20) and 0.29 (0.36-0.22) between the two examiners and the self-assessment (p < 0.0001). CONCLUSIONS: In the studied sample, self-assessment of the pubertal stage should not replace the objective assessment made by trained professionals. Improvement of the self-assessment method may validate its use in population-based studies.

Comparison between objective assessment and self-assessment of sexual maturation in children and adolescents.

OBJECTIVE: To assess the correlations between objective assessment and self-assessment of sexual maturation in the outpatient and school settings. METHODS: Three hundred and nineteen individuals, 178 (96 boys and 82 girls) from an outpatient clinic and 141 (73 boys and 68 girls) from public schools (8.3-18.7 years), of whom 73 individuals (39 girls an d 34 boys) had a body mass index above the 85th percentile, according to 2000 CDC Growth Chart, were analyzed. All of them were examined sequentially and individually by two trained physicians after a written consent form was signed by parents or surrogates, and then submitted to self-assessment using pictures (Tanner stages). Kappa coefficients between examiners and the self-assessment were calculated based on the collected data. A p value <5%was established as statistically significant. RESULTS: No significant difference was observed between correlations obtained from the outpatient clinic and schools, and both groups were combined for analysis. The correlations obtained by examiners were significantly higher than those from self-assessment, with a kappa coefficient (and confidence interval) of 0.75 (0.8-0.69) for breasts/genitals across examiners against 0.27 (0.34-0.20) and 0.29 (0.36-0.22) between the two examiners and the self-assessment (p < 0.0001). CONCLUSIONS: In the studied sample, self-assessment of the pubertal stage should not replace the objective assessment made by trained professionals. Improvement of the self-assessment method may validate its use in population-based studies.

Trypanosoma cruzi: immunoglobulin isotype profiles during the acute phase of canine experimental infection with metacyclic or blood trypomastigotes.

A detailed investigation has been carried out about the serological profiles of groups of dogs experimentally infected with metacyclic (MT) or blood (BT) trypomastigotes of Berenice-78 Trypanosoma cruzi strain. Peripheral blood was collected from infected dogs and uninfected controls, weekly during 35 days following the acute phase of infection, and immunoglobulin profiles were determined by ELISA. Dogs infected with BT exhibited unaltered levels of IgG2, increases in IgM, IgE, IgA, IgG and IgG1. In contrast, dogs infected with MT presented unaltered levels of IgE and IgG1 and an increase in IgM, IgA, IgG and IgG2 levels. Compared with the MT group, animals infected with BT showed significant increases in IgM on days 7, 14 and 28, in IgA on days 7, 14 and 21, in IgE on days 7 and 14, in IgG on days 14 and 28, and in IgG1 on days 7, 14 and 21. Parasitemia levels of the infected animals were measured over the same time period. No correlations were found between the immunoglobulin profiles and the parasitemia levels. The results demonstrated that the inoculum source (BT or MT) influence the immunoglobulin isotype profile that may drive distinct outcome of acute canine Chagas disease.

Persistence of PCR-positive tissue in benznidazole-treated mice with negative blood parasitological and serological tests in dual infections with Trypanosoma cruzi stocks from different genotypes.

OBJECTIVES: To assess different methodologies to better define an early post-therapeutic cure criterion after benznidazole treatment in BALB/c mice following mixed infection with dual Trypanosoma cruzi genotypes. METHODS: According to the classical cure criteria, animals were classified as treated not cured (TNC = 76.4%), treated cured (TC = 12.5%) and dissociated (DIS = 11.1%) using parasitological [fresh blood examination (FBE), blood culture (BC) and blood PCR] and serological methods [conventional serology (CS-ELISA) and non-conventional serology (NCS-FC-ALTA)]. Tissues were also evaluated by PCR. RESULTS: FBE was able to detect patent parasitaemia in only 18.1% of TNC and therapeutic failure was detected in 79.1% and 97.2% of TNC by BC and blood PCR, respectively. CS-ELISA should not be used before 3 months after treatment since it may lead to false-negative results. At 3 months after treatment with benznidazole, NCS-FC-ALTA was more efficient for categorizing the groups of treated mice. In the TNC group, although a decreased frequency of PCR-positive tissue was observed in several host tissues, increased positivity was also observed, despite the T. cruzi genotype combination. All TC animals presented at least two positive tissue-PCR results. CONCLUSIONS: Our results confirm that NSC-FC-ALTA and blood PCR are the most suitable methods to early detect therapeutic failure in acute murine T. cruzi infection. Additionally, our data show that BC positivity is highly dependent upon the T. cruzi genotype combination. Moreover, our findings demonstrated that PCR tests performed on tissues from animals considered cured after benznidazole treatment still detected T. cruzi DNA, most probably indicating residual infection.