Biology and External Morphology of the Immature Stages of Dirphia moderata Bouvier (Lepidoptera: Saturniidae: Hemileucinae) in Anacardium occidentale L / Biologia e Morfologia Externa dos Estágios Imaturos de Dirphia moderata Bouvier (Lepidoptera: Saturniidae: Hemileucinae) em Anacardium occidentale L

Abstract The genus Dirphia Hübner, [1819] presents Neotropical distribution. This genus, besides causing accidents of interest in public health, is a potential defoliator pest of cultivated plants, among them the cashew tree (Anacardium occidentale L.), a crop of great economic importance. This study describes the biology, external morphology of the immature stages of Dirphia moderata Bouvier, 1929 including the first instar larva chaetotaxy. The egg stage had an average duration of 15 days, larval stage 45 days and pupa 60 days, totaling 120 days in average temperature of 28.8 °C and 59.5% relative humidity. Six instars were confirmed by the Dyar rule, with a growth rate of cephalic capsule K = 1.4 times per ínstar. The results found in the first ínstar chaetotaxy follow the general pattern known for Hemileucinae.

Resumo O gênero Dirphia Hübner, [1819] apresenta distribuição Neotropical. Esse gênero, além de causar acidentes de interesse na saúde pública, é uma potencial praga desfolhadora de plantas cultivadas, entre elas o cajueiro (Anacardium occidentale L.), cultura de grande importância econômica. Esse estudo descreve a biologia e morfologia externa dos estágios imaturos de Dirphia moderata Bouvier, 1929 incluindo a quetotaxia da larva de primeiro instar. O estágio de ovo teve duração média de 15 dias, estágio larval de 45 dias e pupa de 60 dias, totalizando 120 dias em temperatura média de 28,8 °C e 59,5% de umidade relativa. Seis ínstares foram confirmados pela regra de Dyar, com razão de crescimento da cápsula cefálica K = 1,4 vezes a cada ínstar. Os resultados encontrados na quetotaxia do primeiro instar segue o padrão geral conhecido para Hemileucinae.

Biology and External Morphology of the Immature Stages of Dirphia moderata Bouvier (Lepidoptera: Saturniidae: Hemileucinae) in Anacardium occidentale L.

The genus Dirphia Hübner, [1819] presents Neotropical distribution. This genus, besides causing accidents of interest in public health, is a potential defoliator pest of cultivated plants, among them the cashew tree (Anacardium occidentale L.), a crop of great economic importance. This study describes the biology, external morphology of the immature stages of Dirphia moderata Bouvier, 1929 including the first instar larva chaetotaxy. The egg stage had an average duration of 15 days, larval stage 45 days and pupa 60 days, totaling 120 days in average temperature of 28.8 °C and 59.5% relative humidity. Six instars were confirmed by the Dyar rule, with a growth rate of cephalic capsule K = 1.4 times per ínstar. The results found in the first ínstar chaetotaxy follow the general pattern known for Hemileucinae.

p27kip1 as a key regulator of endometriosis.

p27kip1 as a key regulator of endometriosis Gonçalves GA p27kip1 is a cyclin-dependent kinase (CDK) inhibitor whose specific late G1 destruction allows progression of the cell across the G1/S boundary. There is a direct relationship between low level of p27 and rapid proliferation occurring in several benign states and in many malignances. In the glandular cells of the normal endometrium, the level of p27kip1 is exceedingly low during the proliferative phase, whereas it is markedly increased during the secretory phase. The expression of p27kip1 in endometriosis is very low but has been found to increase following treatment with progesterone. However, estrogen exposure is considered as a major risk factor in developing endometrial cancer. Endometriosis endometrial cells cultures have also lower levels of p27kip1 compared to heath endometrial cells cultures and restore the cell cycle balance when transduced with an adenoviral vector carring the p27kip1 coding gene (Adp27EGFP). More uniform and rigorous studies are required to confirm these and additional markers utility in a diagnostic and possible treatment panel. As a major clinical priority is to determine which lesions can be treated medically and which require surgical intervention, focusing future studies on markers that distinguish response to hormone therapy or are involved in hormone regulation, will be important future considerations. The goal of this highlight review is to provide a broad overview of the advancements in studies about endometriosis mainly correlating the cytokine p27kip1 expression with the diagnostic and disease treatment.

p27kip1 overexpression regulates IL-1ß in the microenvironment of stem cells and eutopic endometriosis co-cultures.

Endometriosis is a gynecological benign chronic disease defined as the growth of endometrial glands and stroma in extra-uterine sites, most commonly implanted over visceral and peritoneal surfaces within the female pelvis causing inflammatory lesions. It affects around 10% of the female population and is often accompanied by chronic pelvic pain, adhesion formation and infertility. Therefore, endometriosis could be considered a "social disease", since it affects the quality of life, reproductivity and also has a socio-economic impact. The expression of cell cycle and inflammatory proteins is modified in the endometriotic tissues. Immunostaining of glandular and stromal cells in endometrial biopsies obtained from patients with endometriosis compared with those of healthy control demonstrated that endometriotic tissues have lower levels of p27kip1 protein. Endometriosis endometrial cells cultures have also lower levels of p27kip1 compared to health endometrial cells cultures and restore the cell cycle balance when transduced with an adenoviral vector carring the p27kip1 coding gene (Adp27EGFP). The low levels of p27kip1 are related to the S phase in the cell cycle, whereas higher levels lead to a G1 cell cycle arrest. The inflammatory cytokine IL-1ß was recently identified as another key protein in the endometriosis proliferation. This cytokine has elevated levels during the proliferative and secretory phases of the menstrual cycle. In endometriosis endometrial cells cultures the IL-1ß stimulates the production of IL-6 and IL-8, increasing the cell proliferation and reducing the apoptosis and Bax expression in these cells. According to these remarks, this work aims to evaluate the inflammatory effects in vitro, but more next to what happens in a woman's body, associating endometrial cells with stem cells, thus mimicking the endometrial microenvironment, with gene therapy using Adp27, notoriously known as controller cell cycle, apoptosis and potent modulator of VEGF expression.

Biology and External Morphology of the Immature Stages of Dirphia moderata Bouvier (Lepidoptera: Saturniidae: Hemileucinae) in Anacardium occidentale L

Abstract The genus Dirphia Hübner, [1819] presents Neotropical distribution. This genus, besides causing accidents of interest in public health, is a potential defoliator pest of cultivated plants, among them the cashew tree (Anacardium occidentale L.), a crop of great economic importance. This study describes the biology, external morphology of the immature stages of Dirphia moderata Bouvier, 1929 including the first instar larva chaetotaxy. The egg stage had an average duration of 15 days, larval stage 45 days and pupa 60 days, totaling 120 days in average temperature of 28.8 °C and 59.5% relative humidity. Six instars were confirmed by the Dyar rule, with a growth rate of cephalic capsule K = 1.4 times per ínstar. The results found in the first ínstar chaetotaxy follow the general pattern known for Hemileucinae.

Resumo O gênero Dirphia Hübner, [1819] apresenta distribuição Neotropical. Esse gênero, além de causar acidentes de interesse na saúde pública, é uma potencial praga desfolhadora de plantas cultivadas, entre elas o cajueiro (Anacardium occidentale L.), cultura de grande importância econômica. Esse estudo descreve a biologia e morfologia externa dos estágios imaturos de Dirphia moderata Bouvier, 1929 incluindo a quetotaxia da larva de primeiro instar. O estágio de ovo teve duração média de 15 dias, estágio larval de 45 dias e pupa de 60 dias, totalizando 120 dias em temperatura média de 28,8 °C e 59,5% de umidade relativa. Seis ínstares foram confirmados pela regra de Dyar, com razão de crescimento da cápsula cefálica K = 1,4 vezes a cada ínstar. Os resultados encontrados na quetotaxia do primeiro instar segue o padrão geral conhecido para Hemileucinae.

p27kip1 overexpression regulates VEGF expression, cell proliferation and apoptosis in cell culture from eutopic endometrium of women with endometriosis.

We hypothesized that p27(kip1) overexpression can regulate endometriosis cell proliferation, apoptosis and vascular endothelial growth factor (VEGF) expression in the endometrium. The overexpression of p27(kip1) was obtained by transduction of p27(kip1) in primary cultures of endometrium obtained from women with endometriosis tissue with gene therapy technology. First generation bicistronic adenovirus: AdCMVhp27IRESEGFP (Adp27) and AdCMVNull (AdNull) were engineered in order to induce p27(kip1) expression in endometrial cells primary culture. The effect of p27(kip1) overexpression was elucidated through the cell proliferation evaluation and the expression of the cell cycle-related proteins p16, p21, p27, and p53. Cell cycle and apoptosis in endometrial cells from women with and without endometriosis were also evaluated. The VEGF levels were evaluated 1 and 7 days after transduction. The experiments were performed using Immunofluorescence stainings and flow cytometry technique. The cell proliferation statistically diminished markedly following p27(kip1) overexpression in the endometriosis group. This process was accompanied, however, by a statistically significant modulation of the cell cycle-related proteins p16, p21, p27 and p53 markedly increase following p27(kip1) overexpression in the endometriosis group (p < 0.001) and an increase in apoptotic cells was observed. In the endometriosis group, significant downregulation of VEGF expression was observed 7 days after p27(kip1) overexpression, attaining levels strikingly similar to those observed in the control endometrial cells. The findings of this study showed a link between the cell cycle control protein (p27(kip1)) and angiogenesis (VEGF). Our results, also reinforces the background of endometrial dysfunction as part of the origin of endometriosis. We believe that better knowledge of endometrium milieu and the establishment of the link between different, previously describe, altered pathways in this tissue can facilitate future genetic cell therapy.

Immune response of broiler chickens immunized orally with the recombinant proteins flagellin and the subunit B of cholera toxin associated with Lactobacillus spp.

This study investigated the immune response of broiler chickens with oral treatment of a Lactobacillus spp. pool (PL) associated with microencapsulated recombinant proteins flagellin (FliC) and the subunit B of cholera toxin (CTB). Immune responses were evaluated by measuring IgA from intestinal fluid, serum IgY, and immunostaining of CD8(+) T lymphocytes present in the cecum. The evaluations were performed on d 0, 7, 14, 21, and 28 posttreatment. A significant increase (P < 0.05) was observed in IgA levels in all immunized groups, especially 3 wk after immunization. Treatments 2 (recombinant CTB) and 3 (recombinant FliC+CTB) showed the highest concentrations. Similarly, serum concentrations IgY (µg/mL) increased along the experiment, and the means for treatments 2 and 3 showed significant differences (P < 0.05) compared with controls, reaching concentrations of 533 and 540 µg/mL, respectively. The number of CD8(+) T lymphocytes in all treatments greatly differed (P < 0.05) compared with the negative control at 21 d posttreatment. However, only treatment 2 (recombinant CTB), 4 (PL), and 5 (recombinant FliC+ recombinant CTB + PL) remained significantly (P < 0.05) different from the control at 28 d posttreatment. Thus, it is concluded that the microencapsulated recombinant proteins administered orally to broiler chickens are capable of stimulating humoral and cellular immune response, and the combinations of these antigens with Lactobacillus spp. can influence the population of CD8(+) T cells residing in the cecum.

Modelling uncertainty estimation for the determination of aflatoxin M(1) in milk by visual and densitometric thin-layer chromatography with immunoaffinity column clean-up.

The uncertainty of aflatoxin M(1) concentration in milk, determined by thin-layer chromatography (TLC) with visual and densitometric quantification of the fluorescence intensities of the spots, was estimated using the cause-and-effect approach proposed by ISO GUM (Guide to the expression of uncertainty in measurement) following its main four steps. The sources of uncertainties due to volume measurements, visual and densitometric TLC calibration curve, allowed range for recovery variation and intermediary precision to be taken into account in the uncertainty budget. For volume measurements the sources of uncertainties due to calibration, resolution, laboratory temperature variation and repeatability were considered. For the quantification by visual readings of the intensity of the aflatoxin M(1) in the TLC the uncertainty arising from resolution calibration curves was modelled based on the intervals of concentrations between pairs of the calibration standard solutions. The uncertainty of the densitometric TLC quantification arising from the calibration curve was obtained by weighted least square (WLS) regression. Finally, the repeatability uncertainty of the densitometric peak areas or of the visual readings for the test sample solutions was considered. For the test samples with aflatoxin M(1) concentration between 0.02 and 0.5 µg l(-1), the relative expanded uncertainties, with approximately 95% of coverage probability, obtained for visual TLC readings were between 60% and 130% of the values predicted by the Horwitz model. For the densitometric TLC determination they were about 20% lower. The main sources of uncertainties in both visual and densitometric TLC quantification were the intermediary precision, calibration curve and recovery. The main source of uncertainty in the calibration curve in the visual TLC analysis was due to the resolution of the visual readings, whereas in the densitometric analysis it was due to the peak areas of test sample solutions followed by the intercept and slope uncertainties of the calibration line.

Co-occurrence of aflatoxins B1, B2, G1 and G2, ochratoxin A, zearalenone, deoxynivalenol, and citreoviridin in rice in Brazil.

A total of 230 samples of processed rice and its sub-products or derived products were analysed to establish the co-occurrence of several mycotoxins. Samples were analysed in the period 2007-2009 due to the outbreak of beriberi associated with the consumption of rice stored in inappropriate conditions in Brazil. According to data from the Ministry of Health, 323 cases of disease were registered in 2006, of which at least 47 cases resulted in death. The occurrence of total aflatoxin (AFT) (aflatoxin B(1) + B(2) + G(1) + G(2)), ochratoxin A (OTA), zearalenone (ZON), deoxynivalenol (DON), and citreoviridin (CTV) was 58.7%, 40.0%, 45.2%, 8.3% and 22.5%, respectively. From 166 rice samples analysed, 55% had levels <0.11 µg kg(-1) for AFT. For OTA and ZON, of 165 rice samples analysed, 28% and 29% were contaminated with levels from 0.20 to 0.24 µg kg(-1) and from 3.6 to 290.0 µg kg(-1), respectively. One sample (0.6%) was contaminated with 4872.0 µg kg(-1) of ZON. A total of 91% of rice samples (n = 165) did not contain detectable DON (<30.00 µg kg(-1)), although the highest level of contamination was found to be 244 µg kg(-1). From the total of 65 samples analysed, 94% had no detectable CTV (<0.9 µg kg(-1)), with a range from 0.9 to 31.1 µg kg(-1) in 6% of the samples. The highest levels of contamination were found in rice sub-products or derived products from the husk and rice bran. Co-occurrence was observed for AFT and ZON in 17.0%, AFT and OTA in 24.2%, AFT and CTV in 6.2%, OTA and CTV in 4.6%, and ZON and CTV in 3.1%. These fractions were also the major contributors for the co-occurrence. The results found show the necessity of monitoring rice production.

Susceptibilidade antimicrobiana de amostras de Escherichia coli isoladas de frango industrial (Gallus gallus domesticus - Linnaeus, 1758) com colibacilose / Antimicrobial susceptibility of samples of Escherichia coli isolated from industrial chickens (Gallus gallus domesticus - Linnaeus, 1758) with colibacillosis

O presente estudo avaliou a susceptibilidade de amostras de Escherichia coli isoladas de quadros de colissepticemia aviária da região centro-oeste paulista pelo Laboratório de Ornitopatologia da FMVZ-UNESP/Botucatu, SP. Constatou-se um grande número de amostras multirresistentes aos antibióticos testados, onde as drogas menos efetivas foram sulfonamida e tetraciclina. Todas as amostras mostraram-se sensíveis a norfloxacina e gentamicina.

The present study evaluated the susceptibility of Escherichia coli samples isolated from poultry with colisepticemia in midwestern São Paulo State at the Ornitopathology Laboratory of FMVZ-UNESP/Botucatu, SP. A large number of samples were found that were multi-resistant to the antibiotics tested, the less effective drugs being sulfonamide and tetracycline. All samples were susceptible to norfloxacin and gentamicin.

Intramyocardial transplantation of fibroblasts expressing vascular endothelial growth factor attenuates cardiac dysfunction.

In this study, we analyzed whether transplantation of cardiac fibroblasts (CFs) expressing vascular endothelial growth factor (VEGF) mitigates cardiac dysfunction after myocardial infarction (MI) in rats. First, we observed that the transgene expression lasts longer (45 vs 7 days) when fibroblasts are used as vectors compared with myoblasts. In a preventive protocol, induction of cardiac neovascularization accompanied by reduction in myocardial scar area was observed when cell transplantation was performed 1 week before ischemia/reperfusion and the animals analyzed 3 weeks later. Finally, the therapeutic efficacy of this approach was tested injecting cells in a fibrin biopolymer, to increase cardiac retention, 24 h post-MI. After 4 weeks, an increase in neovascularization and a decrease in myocardial collagen were observed only in rats that received cells expressing VEGF. Basal indirect or direct hemodynamic measurements showed no differences among the groups whereas under pharmacological stress, only the group that received cells expressing VEGF showed a significant reduction in end-diastolic pressure and improvement in stroke volume and cardiac work. These results indicate that transplantation of CFs expressing VEGF using fibrin biopolymer induces neovascularization and attenuates left ventricle fibrosis and cardiac dysfunction in ischemic heart.

Intestinal protozoan parasites with zoonotic potential in birds.

The aim of this study was to evaluate the occurrence of potentially zoonotic intestinal protozoan infections in exotic and wildlife Brazilian birds. Fecal samples from 207 birds of 45 species were examined. Infections by Balantidium sp., Entamoeba sp., and Blastocystis sp. were observed in 17 individuals (8.2%) of Gnorimopsar chopi, Oryzoborus angolensis, Sporophila caerulescens, Ramphastos toco, Aratinga leucophtalmus, and Pavo cristatus.

A tank-to-wheel analysis tool for energy and emissions studies in road vehicles.

Currently, oil based fuels are the primary energy source of road transport. The growing need for oil independence and CO(2) mitigation has lead to the increasing importance of alternative fuel usage. CO(2) is produced not only as the fuel is used in the vehicle (tank-to-wheel contribution), but also upstream, from the fuel extraction to the refueling station (well-to-tank contribution), and the life cycle of the fuel production (well-to-wheel contribution) must be considered in order to analyse the global impact of the fuel utilization. A road vehicle tank-to-wheel analysis tool that may be integrated with well-to-tank models was developed in the present study. The integration in a demonstration case study allowed to perform a life cycle assessment concerning the utilization of diesel and natural gas fuels in a specific network line of a bus transit company operating in the city of Porto, Portugal.