Fetal-Maternal Interactions in the Synepitheliochorial Placenta Using the eGFP Cloned Cattle Model

Background: To investigate mechanisms of fetal-maternal cell interactions in the bovine placenta, we developed a model of transgenic enhanced Green Fluorescent Protein (t-eGFP) expressing bovine embryos produced by nuclear transfer (NT) to assess the distribution of fetal-derived products in the bovine placenta. In addition, we searched for male specific DNA in the blood of females carrying in vitro produced male embryos. Our hypothesis is that the bovine placenta is more permeable to fetal-derived products than described elsewhere. Methodology/Principal Findings: Samples of placentomes, chorion, endometrium, maternal peripheral blood leukocytes and blood plasma were collected during early gestation and processed for nested-PCR for eGFP and testis-specific Y-encoded protein (TSPY), western blotting and immunohistochemistry for eGFP detection, as well as transmission electron microscopy to verify the level of interaction between maternal and fetal cells. TSPY and eGFP DNA were present in the blood of cows carrying male pregnancies at day 60 of pregnancy. Protein and mRNA of eGFP were observed in the trophoblast and uterine tissues. In the placentomes, the protein expression was weak in the syncytial regions, but intense in neighboring cells on both sides of the fetal-maternal interface. Ultrastructurally, our samples from t-eGFP expressing NT pregnancies showed to be normal, such as the presence of interdigitating structures between fetal and maternal cells. In addition, channels-like structures were present in the trophoblast cells. Conclusions/Significance: Data suggested that there is a delivery of fetal contents to the maternal system on both systemic and local levels that involved nuclear acids and proteins. It not clear the mechanisms involved in the transfer of fetal-derived molecules to the maternal system. This delivery may occur through nonclassical protein secretion; throughout transtrophoblastic-like channels and/or by apoptotic processes previously described. In conclusion, the bovine synepitheliochorial placenta displays an intimate fetal-maternal interaction, similar to other placental types for instance human and mouse. © 2013 Pereira et al.

Identificação e análise de polimorfismos no gene que codifica a Lectina ligante de Manose (MBL) na espécie Canis lúpus familiaris

Pós-graduação em Biologia Geral e Aplicada - IBB

Enxertos venosos ao avesso e normal, preenchidos com plasma rico em plaquetas em nervo misto de rato

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo experimental sobre a resposta imunológica em infecções seqüenciais pelo vírus Dengue 3 e pelo vírus Dengue 2 em primatas não humanos da espécie Callithrix penicillata

A dengue é causada pelo Vírus da dengue (VDEN - Flaviviridae, Flavivirus) e é considerada a arbovirose mais difundida no mundo. Atualmente, não há um animal que sirva como modelo para estudos sobre a patogênese do VDEN. Para investigar a susceptibilidade da espécie Callithrix penicillata ao VDEN foram inoculados amostras do VDEN-3 e do VDEN-2 de isoladas de casos humanos fatais. Para tal, vinte e dois animais foram infectados com VDEN-3 (3,23 × 103 PFU/mL) e, sessenta dias após a infecção (d.p.i.), 11 deles foram secundariamente infectados com VDEN-2 (4,47 × 104 PFU/mL). Sangue, plasma e soro, foram coletados diariamente durante os primeiros sete dias e em 15, 20, 45 e 60 d.p.i.. Foram investigados a produção de anticorpos IgM e inibidores da hemaglutinação, assim como foram análisados parâmetros hematologicos e bioquímicos e o perfil sérico de citocinas (IL-6, TNF-, IL-2, IFN-α, IL-4 e IL-5). A infecção primária (VDEN-3) revelou anticorpos IgM (15- 20 d.p.i.), anticorpos IH (15-60 d.p.i.), aumento dos níveis de TNF-α e IFN-γ e diminuição da IL-5. Na infecção secundária (VDEN-2), foi detectado anticorpos IgM (15-20 d.p.i.), anticorpos IH (15-60 d.p.i.) e diminuição dos níveis de IL-6, TNF-α, de IFN-γ e IL-5. Além disso, foi observado em ambas infecções leucopenia, neutropenia, linfocitopenia, monocitopenia, trombocitopenia, e aumentou da AST. O aumento dos níveis de INF-γ e TNF- α indicaram a ativação da resposta inflamatória, com a diferenciação para a resposta celular e supressão da proliferação de citocinas características da resposta humoral. A presença de anticorpos neutralizantes pode ter ocasionado a supressão da resposta inflamatória na infecção secundária o que pode ter levado a ausência de sinais de Febre Hemorrágica do Dengue/Sindrome do Choque do Dengue (FHD/SCD) durante a infecção secundária (VDEN- 2). Os resultados indicam que o primatas não humanos da espécie Callithrix penicillata demonstraram susceptibilidade à cepas de VDEN de origem humana, sugerindo que esses animais são um bom modelo para estudos da resposta imunológica da Febre do Dengue (FD), assim como para a avaliação de uma vacina tetravalente.

Conversão in vitro e in vivo do pró-fármaco hidroximetilnitrofural, candidato a fármaco antichagásico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Blood clot stabilization on root dentin conditioned by the combination of tetracycline and EDTA

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detecção do HPV e EBV por nPCR em líquen plano bucal e tecido normal de cavidade bucal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A simple method based on ICP-MS for estimation of background levels of arsenic, cadmium, copper, manganese, nickel, lead, and selenium in blood of the Brazillian population

Throughout the world, biomonitoring has become the standard for assessing exposure of individuals to toxic elements as well as for responding to serious environmental public health problems. However, extensive biomonitoring surveys require rapid and simple analytical methods. Thus, a simple and high-throughput method is proposed for the determination of arsenic (As), cadmium (Cd), copper (Cu), manganese (Mn), nickel (Ni), lead (Pb), and selenium (Se) in blood samples by using inductively coupled plasma–mass spectrometry (ICPMS). Prior to analysis, 200 ml of blood samples was mixed with 500 ml of 10% v/v tetramethylammonium hydroxide (TMAH) solution, incubated for 10 min, and subsequently diluted to 10 ml with a solution containing 0.05% w/v ethylenediamine tetraacetic acid (EDTA) + 0.005% v/v Triton X-100. After that, samples were directly analyzed by ICP-MS (ELAN DRC II). Rhodium was selected as an internal standard with matrix-matching calibration. Method detection limits were 0.08, 0.04, 0.5, 0.09, 0.12, 0.04, and 0.1 mg//L for As, Cd, Cu, Mn, Ni, Pb, and Se, respectively. Validation data are provided based on the analysis of blood samples from the trace elements inter-\comparison program operated by the Institut National de Santé Publique du Quebec, Canada. Additional validation was provided by the analysis of human blood samples by the proposed method and by using electrothermal atomic absorption spectrometry (ETAAS). The method was subsequently applied for the estimation of background metal blood values in the Brazilian population. In general, the mean concentrations of As, Cd, Cu, Mn, Ni, Pb, and Se in blood were 1.1, 0.4, 890, 9.6, 2.1, 65.4, and 89.3 mg/L, respectively, and are in agreement with other global populations. Influences of age, gender, smoking habits, alcohol consumption, and geographical variation on the values were also considered. Smoking habits influenced the levels of Cd in blood. The levels of Cu, Mn, and Pb were significantly correlated with gender, whereas Cu and Pb were significantly correlated with age. There were also interesting differences in Mn and Se levels in the population living in the north of Brazil compared to the south.

Otimização de um método analítico para determinação de ácidos graxos em plasma sanguíneo de rato

This work has as main theme optimize the method of determination of fatty acids such as methyl octanoate, methyl palmitate, methyl stearate, methyl oleate, methyl linoleate and methyl linolenate in blood plasma samples from mice. The method proved to be very suitable for the analysis, in which we obtained the following linear coefficients: 0.9992, 0.9989, 0.9996, 0.9995, 0.9999 for methyl linoleate acid, methyl oleate, methyl palmitate, methyl stearate and methyl octanoate, respectively. Esterification of the samples gave good reading of chromatograms of samples without interfering peaks. The results obtained were as expected from the diet of mices

Plasma sanguíneo desidratado na alimentação da Tilápia-do- Nilo

Pós-graduação em Zootecnia - FMVZ

Hemocompatibility of poly(epsilon-caprolactone) lipid-core nanocapsules stabilized with polysorbate 80-lecithin and uncoated or coated with chitosan

The hemocompatibility of nanoparticles is of critical importance for their systemic administration as drug delivery systems. Formulations of lipid-core nanocapsules, stabilized with polysorbate 80-lecithin and uncoated or coated with chitosan (LNC and LNC-CS), were prepared and characterized by laser diffraction (D[4,3]: 129 and 134 nm), dynamic light scattering (119 nm and 133 nm), nanoparticle tracking (D50: 124 and 139 nm) and particle mobility (zeta potential: -15.1 mV and + 9.3 mV) analysis. In vitro hemocompatibility studies were carried out with mixtures of nanocapsule suspensions in human blood at 2% and 10% (v/v). The prothrombin time showed no significant change independently of the nanocapsule surface potential or its concentration in plasma. Regarding the activated partial thromboplastin time, both suspensions at 2% (v/v) in plasma did not influence the clotting time. Even though suspensions at 10% (v/v) in plasma decreased the clotting times (p < 0.05), the values were within the normal range. The ability of plasma to activate the coagulation system was maintained after the addition of the formulations. Suspensions at 2% (v/v) in blood showed no significant hemolysis or platelet aggregation. In conclusion, the lipid-core nanocapsules uncoated or coated with chitosan are hemocompatible representing a potential innovative nanotechnological formulation for intravenous administration. (C) 2012 Elsevier B. V. All rights reserved.

Determination of trace elements in bovine semen samples by inductively coupled plasma mass spectrometry and data mining techniques for identification of bovine class

The reproductive performance of cattle may be influenced by several factors, but mineral imbalances are crucial in terms of direct effects on reproduction. Several studies have shown that elements such as calcium, copper, iron, magnesium, selenium, and zinc are essential for reproduction and can prevent oxidative stress. However, toxic elements such as lead, nickel, and arsenic can have adverse effects on reproduction. In this paper, we applied a simple and fast method of multi-element analysis to bovine semen samples from Zebu and European classes used in reproduction programs and artificial insemination. Samples were analyzed by inductively coupled plasma spectrometry (ICP-MS) using aqueous medium calibration and the samples were diluted in a proportion of 1:50 in a solution containing 0.01% (vol/vol) Triton X-100 and 0.5% (vol/vol) nitric acid. Rhodium, iridium, and yttrium were used as the internal standards for ICP-MS analysis. To develop a reliable method of tracing the class of bovine semen, we used data mining techniques that make it possible to classify unknown samples after checking the differentiation of known-class samples. Based on the determination of 15 elements in 41 samples of bovine semen, 3 machine-learning tools for classification were applied to determine cattle class. Our results demonstrate the potential of support vector machine (SVM), multilayer perceptron (MLP), and random forest (RF) chemometric tools to identify cattle class. Moreover, the selection tools made it possible to reduce the number of chemical elements needed from 15 to just 8.

Comparison of venous plasma glycemia and capillary glycemia for the screening of type 2 diabetes mellitus in the Japanese-Brazilian community of Mombuca (Guatapará-SP)

Abstract Background To identify the most appropriate cut-off points of fasting glycemia for the screening of diabetes mellitus type 2 (DM2) with the comparison of the properties of capillary glycemia (CG) and venous blood plasma glycemia (PG) in a population of Japanese origin from the community of Mombuca, Guatapará - SP, Brazil. Methods This was a population-based descriptive cross-sectional study conducted on a sample of 131 individuals of both genders aged 20 years or more (66.8% of the target population). CG was measured with a glucometer in a blood sample obtained from the fingertip and PG was determined by an enzymatic method (hexokinase) in venous blood plasma, after a 10-14 hour fast in both cases. Data were analyzed by the receiver operating characteristic (ROC) curve in order to identify the best cut-off point for fasting glycemia (CG and PG) for the diagnosis of DM, using the 2-hour plasma glycemia > 200 mg/dl as gold - standard. Results The ROC curve revealed that the best cut-off point for the screening of DM was 110 mg/dl for CG and 105 mg/dl for PG, values that would optimize the relation between individuals with positive and false-positive results. The area under the ROC curve was 0.814 for CG (p < 0.01) and 0.836 for PG (p < 0.01). Conclusions The cut-off points of 105 mg/dl(5.8 mmol/l) for PG and of 110 mg/dl(6.1 mmol/l) for CG appear to be the most appropriate for the screening of DM2 in the population under study, with emphasis on the fact that the value recommended for CG is 5 mg/dl higher than that for PG, in contrast to WHO recommendations.

Estresse oxidativo no plasma sanguíneo de indivíduos submetidos ao esforço físico agudo seguido de crioimersão corporal

O objetivo deste estudo foi analisar a influência da crioimersão corporal (CIC) imediata ao esforço físico agudo no estresse oxidativo (EOx) no plasma sanguíneo. Participaram do presente estudo 12 homens, com idade média de 22±1 anos, submetidos ao teste de esforço físico intenso em esteira, seguido de CIC em um tanque com água a 10ºC durante 10 minutos contínuos. Do repouso ao final da CIC, os indivíduos foram monitorados através de alguns parâmetros como: o índice de percepção subjetiva do esforço (IPE) expresso conforme escala de Borg, frequência cardíaca (FC), pressão arterial (PA) e temperatura corporal (TC) através da temperatura timpânica. A análise morfológica do EOx plasmático foi realizada de acordo com o método denominado Morfologia Óptica do Estresse Oxidativo no Plasma (MEOP), utilizando-se gotas de sangue capilar. Observou-se uma significativa elevação (p<0,01) no grau do estresse oxidativo plasmático após a realização do esforço físico, em relação ao respectivo grau em repouso. Porém, esta elevação no grau do EOx foi significativamente reduzida (p<0,001) em função da CIC. Embora sejam necessários mais estudos científicos com o MEOP, concluiu-se que, para o presente estudo, este teste mostrou-se viável. Os dados encontrados no presente estudo sugerem que a CIC em água a 10ºC por 10 minutos imediatos ao esforço físico agudo com intensidade alta, apresenta-se como uma importante conduta fisioterapêutica para a normalização do EOx pós-esforço.

Modulation of plasma levels and percentages of incorporation of ω-3 PUFAs in egg yolk under the influence of supplementation sources rich in omega 3 to diet of laying

Hundred forty-four Shaver White laying hens were used over a 4 week experimental period to investigate the effect of 3% of soybean oil, corn oil (MIL), canola oil, flaxseed oil (LIN), salmon oil (SAL) or tuna and sardine oil (SR/AT) added to the diets, upon the fatty acid egg yolk composition, blood plasma levels and incorporation time of each fatty acid into the egg yolk. Hens were allocated into 72 cages and the experimental design was a 6 x 6 randomized factorial model. Hens fed 3% of different oils, responded with increased polyunsaturated fatty acids omega 3 (ω-3 PUFAs), except for corn oil. The addition of flaxseed, soybean or corn oil into the diet increased the PUFAs levels into the egg yolk and in the blood plasma. Adding tuna and sardine oil into the diet increased the concentration of yolk saturated fatty acids. The levels of ω-3 PUFAs were increased in the tuna and sardine oil treatment, while the flaxseed oil increased the plasma fatty acids. The deposition of 349.28 mg/yolk of a-linolenic fatty acids (ALA) was higher in the group fed LIN, while the higher equal to 157.13 mg DHA/yolk was observed in group SR/AT. In the plasma, deposition increased from 0.33% (MIL) for 6.29% ALA (LIN), while that of DHA increase of 0.47% (MIL) for 4.24% (SAL) and 4.48% (SR/AT) and of 0.98% (MIL) for 6.14% (SR/AT) and 8.44% (LIN) of ω-3 PUFAs. The percentage of EPA into the yolk and plasma was higher for the hens fed 3% tuna and sardine oil diet, as well as the levels of yolk DHA. The concentration of DHA into the plasma was higher for the salmon and tuna/sardine oil treatments. The PUFAs yolk decreased during the first eight days of experiment, while the ω-3 PUFAs increased during the same period. The concentration of ALA increased until ten days of experiment, while the percentage of EPA and DHA increased up to the eighth experimental day