Comparison between a soluble antigen-based ELISA and IFAT in detecting antibodies against Babesia canis in dogs

O presente trabalho estudou um ensaio imunoenzimático (ELISA) indireto para a detecção de anticorpos anti-Babesia canis no soro de cães, tendo a Reação de Imunofluorescência Indireta (RIFI), como teste de referência O antígeno utilizado no ELISA do presente estudo consistiu em uma preparação antigênica solúvel de merozoítas B. canis e as diluições ótimas do antígeno, soros e conjugado foram determinadas por titulação em bloco, utilizando soros de referência positivos e negativos. A preparação antigênica solúvel de B. canis ótima foi de 10 µg.mL-1, com soros de referência positivos e negativos em uma única diluição de 1:100, e conjugado a 1:4.000. Um total de 246 amostras séricas foram colhidas em cães, durante a campanha de vacinação anti-rábica em Jaboticabal, São Paulo, Brasil e a presença de anticorpos anti-B. canis foi avaliada pelo ELISA e RIFI. Nestas condições, a média de absorbância dos soros de referência negativos foi de 0,129 ± 0,025, resultando em um ponto de corte de 0,323 (Nível de ELISA 3) e a média da absorbância dos soros de referência positivos foi de 2,156 ± 1,187. As amostras com sorologia positiva para B. canis por ELISA e RIFI foram 67,89% (n = 167) e 59,35% (n = 146), respectivamente. Os resultados obtidos sugerem que o ELISA descrito revelou-se um teste sorológico eficaz no diagnóstico da babesiose canina.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Chicken serologic response to Salmonella enterica serotype Typhimurium assessed by Elisa

This study evaluated two enzyme-linked immunosorbent assays (ELISA) in the detection of chicken serologic response against Salmonella enterica sorotype Typhimurium. The assays have used as detecting antigen the soluble bacterial proteins of a non-flagellated strain of Salmonella Typhimurium (AgTM), and antibody conjugated to peroxidase or alkaline phosphatase. According to the results, optimal dilutions of antigen (concentration 5.49 mg/mL) and serum samples in both assays were 1:20,000 and 1:1,000, respectively. In such conditions, the ELISA/AgTM was able to detect serological response to Salmonella Typhimurium. Cross-reactions to Salmonella serotypes Gallinarum and Pullorum were seen, but not with other serotypes such as Enteritidis.

Representações sociais do comportamento agressivo do homem sob a ótica da mulher em situação de violência

We aim to understand the social representations of man's aggressive behavior from the perspective of women in situations of domestic violence. This is a descriptive, exploratory and representational study, whose methodological approach falls into the qualitative category. We chose as a scenario for research, by the Reference Center for Citizen Women (CRMC), Natal / RN. The criteria for selection of participants were women who lived/live in situations of domestic violence, with affective or relationship bonding with the assailant, in psychological and emotional positions appropriated to the reality; that are being protected or assisted by the service listed above; whose aggressor is male. We adopted as data collection instruments: questionnaire, Drawing-Story (DE) and a field diary. For analysis of textual data, we decided to use the ALCESTE software conjugated to editing analyze and initial reading. Were investigated 20 women victims of domestic violence, whose author of the attacks was the husband/partner. We identified, from the respondents, that 70% (n = 14) of men with aggressive behavior also had a family history of violence and fragile family relationships. About the physical and emotional condition of the assailant at the time of violence, 50% (n = 10) of these men, regardless the use of alcohol, had often quarrelsome and/or nervous behavior, impatient and unpredictable humor facing a setback, worry or annoyance. Regarding the nature of violence, we observed that women were victims of all types of violence, however, the psychological prevailed in 100% of cases. The corpus "Men" has three classes, whose focuses are, respectively: resignation, denounce and violence/aggression, being possible categorizing them as well: Category 1: The imprisonment of women; Category 2: Violence and its meanings; Category 3: Breaking the violent cycle. We show that the social representations of man s aggressive behavior, from the women in situations of violence, are anchored in the social roles of men in family and society, becoming a dominant model of masculinity. It is aimed, on one hand, from the reproduction of what is already known and/or experienced by male aggressors in the family, as repetitions of behavior. And on the other, present themselves as a state of illness, addiction or psychopathy

Composição química e perfil de ácidos graxos da carne de bovinos de diferentes condições sexuais recebendo silagem de milho e concentrado ou cana-de-açúcar e concentrado contendo grãos de girassol

O objetivo neste trabalho foi avaliar as características químicas e a composição em ácidos graxos do contrafilé (músculo Longissimus) de tourinhos, novilhos e novilhas da raça Canchim. Os animais foram terminados em confinamento com duas dietas experimentais, uma com silagem de milho e concentrado e outra com cana-de-açúcar e concentrado contendo grãos de girassol. Os teores de umidade, proteína e minerais no músculo não diferiram entre as dietas e as condições sexuais dos animais. A carne de novilhos terminados com a dieta com grãos de girassol apresentou maior teor de lipídios (3,31%) em comparação à dos animais de outras categorias. Os animais que receberam a dieta com grãos de girassol apresentaram maiores concentrações de ácido linoléico conjugado (18:2 cis9, trans-11) (0,73%) e ácidos graxos poliinsaturados (8,12%) no músculo, e também relações mais elevadas de ácidos graxos insaturados:saturados (0,93) e ácidos graxos poliinsaturados:saturados (0,16) em comparação àqueles que receberam a dieta convencional, à base de silagem de milho (0,34%; 6,31%; 0,86; e 0,11, respectivamente). A composição em ácidos graxos da carne de bovinos pode ser melhorada com a utilização de cana-de-açúcar e grãos de girassol na dieta dos animais terminados em confinamento.

Composição em ácidos graxos e qualidade da carne de tourinhos Nelore e Canchim alimentados com dietas à base de cana-de-açúcar e dois níveis de concentrado

O objetivo neste trabalho foi avaliar a composição de ácidos graxos e a qualidade do contrafilé (músculo Longissimus lumborum) de tourinhos das raças Nelore e Canchim. Os animais foram terminados em confinamento e alimentados com dietas contendo cana-de-açúcar e dois níveis de concentrado (40 e 60% na matéria seca). Os concentrados foram compostos de grãos de girassol, milho, farelo de soja, levedura seca de cana-de-açúcar, uréia e núcleo mineral. O delineamento experimental foi o inteiramente ao acaso, em esquema fatorial 2 × 2 (grupo genético × nível de concentrado). Não foram observadas diferenças nos teores de umidade, proteína e extrato etéreo da carne. Os animais da raça Nelore apresentaram maiores concentrações de ácido linoléico conjugado (0,52%), ácidos graxos insaturados (46,82%) e também relações mais elevadas de ácidos graxos insaturados:saturados (1,02) e monoinsaturados:saturados (0,86) em comparação aos tourinhos da raça Canchim. Os tourinhos da raça Canchim apresentaram maior intensidade das cores vermelha e amarela no contrafilé e maior luminosidade da gordura de cobertura. Houve interação para força de cisalhamento, que foi menor nos tourinhos Nelore alimentados com 40% de concentrado. Tourinhos da raça Nelore apresentam carne com melhor composição de ácidos graxos na gordura intramuscular do ponto de vista da saúde humana.

Antibody-targeted horseradish peroxidase associated with indole-3-acetic acid induces apoptosis in vitro in hematological malignancies

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

Entre a fé, a obra social e a publicidade: uma análise crítica do discurso da responsabilidade social da Igreja Universal do Reino de Deus

Situated on Applied Linguistics (PENNYCOOK, 1998; MOITA LOPES, 2003, 2006, 2008, 2009), this thesis, which is inscribed in a qualitative-interpretative approach of a critical analysis perspective , lies on the speech of social responsibility and the way like that is employed in seeking for legitimacy and prestige within the neopentecostal brazilian religious field, more specifically of the Universal Church of the Kingdom of God. The general goal of this research is t reflect upon the speech on social responsibility and the rethoric of selfpromotion of the Universal Church through the role of social actors in the making of his her identities, materialized in the newspaper Folha Universal. In order to achieve that, we have conjugated, in this research, social and discursive analysis. On the linguistic-discursive approach, the research is based on the Critical Analysis of Discourse (ACD), specially in Fairclough (2001, 2003, 2006), a proposal that provides theoretical-methodological tools to investigate the language beyond the linguistic structures , that is, the discourse, social practices in which it occurs and more ample structures. Theoretical assumptions were also used of Sistemico-Functional Linguistics(LSF), matching with categories of the Transitivity System of Halliday (1994, 2004), of the forms of representation of social actors in the socioeconomic perspective by Van Leeuwen (1997, 2008) and of the Appraisal system by Martin and White (2005). As we develop the argumentation on thesobre social role of religion in this thesis, we make use of the authors such as Freston (1994), Oro (1997, 2003), Campos (1996, 1999), Mariano (1999), Meneses (2008), among several ones. We have also used a series of concepts and categories coming from the field of communication and marketing on the business social responsability and social marketing . In this area, we take as references the contributions of Bueno (2003), Fossá and Sartoretto (2003) and Zenone (2006). The corpus of the work is framed by news taken at the newspaper Folha Universal, in which are given the social responsibility actions of the church . The timeframe used was on the editions of 2010 thru 2012. Results found at he analysis of the News lead to semantic features of Assessment of Affection, Judgment and Appreciation, many times followed by Gradation, and the Attribution, one of the subsystems of the Attachment, are evidence of positive assessments for the Universal Church and its agents and make up rethorical elements which provide structure for the discourse of the Universal Church at the newspaper Folha Universal consisted of its image (style) of social responsibility . Results show that the most frequente social actors of the discourse are, on one hand, the Universal Church itself and its volunteers, famous (actors, actresses, presenters), politicians and authorities, on the other hand, the population which was helped by the Church social projects . The first group seems to be Always activated, however the second one, most of the time rather passive. These are also represented by assimilation in most of the occurrences, however the other ones by individualization and nomination entitled by honorification, except for the volunteers that are represented either as an individual, or as a group

Preparation of polymeric micelles for use as carriers of tuberculostatic drugs

Purpose: This paper focuses on the characterization of polymeric micelle-forming tuberculostatic prodrugs and the antimycobacterial activity of these prodrugs.Method: By the condensation of hydroxymethylpyrazinamide, isoniazid and rifampin with free carboxyl groups on the copolymer poly(ethyleneglycol)-poly(aspartic acid), micelle-forming carrier-drug conjugates were obtained. These micelles were characterized by dynamic light scattering, to measure the micelle diameter; by acid-base titration, to determine the percentage of carboxylic groups occupied by the tuberculostatic; by Sudan III solubility tests, to estimate the critical micelle concentration (CMC); and visual control and spectrophotometric measurement, to determine the stability of micelles. These micelles were tested in vitro against several Mycobacterium strains.Results: As expected, the size and distribution of the micelle-forming tuberculostatic prodrugs found to be small (78.2nm, 84.2nm and 98.9 nm) while the level of the drug conjugated was high (65.02-85.7%). Furthermore, the micelles were stable in vitro, exhibiting a low level of CMC and stronger antimycobacterial activity than the original drugs.Conclusion: the results demonstrate that polymeric micelles can be used as efficient carriers for drugs, which alone, exhibit undesired pharmacokinetics, poor solubility, and low stability. The synthesized micelle-forming tuberculostatic prodrugs opens a perspective of alternative prodrugs that prolong action and decrease the toxicity of the tuberculostatic drugs of choice.

Controlled Drug Release from Ureasil-Polyether Hybrid Materials

Flexible, transparent, and insoluble urea-cross-linked polyether-siloxane hybrids presenting a tunable drug delivery pattern were prepared using the sol-gel method from PEO (poly(ethylene oxide)) and PPO (poly(propylene oxide)) functionalized at both chain ends with triethoxysilane. Different polyether chain lengths were used to control the urea/siloxane (named ureasil) node density, flexibility, and swellability of the hybrid network. We herein demonstrate that the drug release from swellable hydrophilic ureasil-PEO hybrids can be sustained for some days, whereas that from the unswellable ureasil-PPO hybrids can be sustained for some weeks. This outstanding feature conjugated with the biomedically safe formulation of the ureasil cross-linked polyether-siloxane hybrid widens their scope of application to include the domain of soft and implantable drug delivery devices.

Otimização de ELISA empregando a proteína Tc85-11 e planejamento fatorial

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

Tafonomia de moluscos fósseis do Grupo Bauru (Cretáceo Superior, Bacia Bauru), na região do município de Monte Alto, São Paulo, Brasil

Bivalves mollusks fossils of Bauru Group (Late Cretaceous, Bauru Basin) deposited in scientific collections and collected in outcrops from Monte Alto municipality, São Paulo, are analyzed in their taphonomy. The preservation of recrystallized individual in carbonatic matrix indicates substrate remobilization by unidirectional energetic event in fluvial discharge. The specimens with conjugated valves possess internal sediment similar to the external indicating low exposition to Taphonomical Active Zone, suggesting a bioclastic low time-averaging. The truncate and fragmented posterior portion of specimens from scientific collections is probably related to the incapacity of the taxa to reburrowing the substrate in drowning periods. Both taphonomic patterns corroborate evidences of a fluvial paleoenvironment in the Bauru Group.

Conjugação e validação de controle isotípico IgG1-FITC para uso em citometria de fluxo

Em meados da década de 50 iniciou-se o desenvolvimento da citometria de fluxo, tecnologia que permite verificar características físico-químicas de células ou partículas suspensas em meio fluido. Esta tecnologia utiliza anticorpos monoclonais marcados com fluorocromos como ferramenta de investigação em diversas análises e necessita de controles isotípicos para definição da região negativa (background). Estes controles são constituídos por imunoglobulinas de mesmo isotipo e fluorocromo dos anticorpos testes, sendo o isotiocianato de fluoresceína (FITC) o marcador fluorescente mais utilizado na conjugação de anticorpos. Os controles isotípicos têm como função definir a fluorescência inespecífica (células negativas) e as regiões fluorescentes (células positivas). No presente estudo foi selecionado anticorpo monoclonal murino (AcMm) dirigido contra antígeno eritrocitário canino, produzido no Laboratório de Anticorpos Monoclonais do Hemocentro de Botucatu, o qual reage positivamente com hemácias de cães, mas nunca com leucócitos humanos, tendo, portanto, potencial utilidade como controle negativo em citometria de fluxo. A purificação do AcMm da subclasse IgG1 foi feita por cromatografia de afinidade em Proteína-A Sepharose, e o controle da purificação realizado por eletroforese em géis de ágarose e poliacrilamida (SDS-PAGE). A imunoglobulina purificada foi conjugada ao FITC e filtrado em coluna de Sephadex G-25 para separação das proteínas marcadas e não-marcadas. O AcMm conjugado foi testado contra hemácias de cães, e o êxito da conjugação comprovado por testes de fluorescência, sendo a mediana de positividade de 94,70. Frente a leucócitos humanos a mediana de positividade foi 0,03 contra 0,50 dos reagentes comerciais. Os testes estatísticos não-paramétricos de Wilcoxon e correlação de Spearman comprovaram a eficiência e validam o controle isotípico produzido em comparação aos reagentes comerciais testados.

Serological diagnosis of visceral leishmaniasis by an enzyme immunoassay using protein A in naturally infected dogs

Um ensaio de imunoadsorção enzimática para detecção de anticorpos contra Leishmania chagasi, utilizando antígeno total de formas promastigotas lisados foi desenvolvido. Cinqüenta cães com sintomas clínicos de leishmaniose visceral foram examinados. Esta técnica utilizou anti-IgG de cão conjugado a peroxidase ou proteína A conjugado a peroxidase. Foi verificado que nos animais positivos diagnosticados por exame parasitológico direto o ensaio ELISA utilizando proteína A conjugada a peroxidase (média da densidade óptica ± desvio padrão 2,078 ± 0,631) detecta mais anticorpos do que o sistema utilizando anti-IgG de cão conjugado a peroxidase (média da densidade óptica ± desvio padrão 1,008 ± 0,437), enquanto para os animais negativos o resultado obtido nos dois sistemas de detecção são similares. Esse resultado sugere que o sistema de ELISA utilizando proteína A conjugado a peroxidase pode ser útil na detecção de animais na fase aguda da infecção e desta forma auxiliar na identificação dos animais positivos e no controle desta importante zoonose.

Origin of sensory and autonomic innervation of the rat temporomandibular joint: A retrograde axonal tracing study with the fluorescent dye fast blue

Previous studies that have used retrograde axonal tracers (horseradish peroxidase alone or conjugated with wheat germ agglutinin) have shown that the temporomandibular joint (TMJ) is supplied with nerve fibers originating mainly from the trigeminal ganglion, in addition to other sensory and sympathetic ganglia. The existence of nerve fibers in the TMJ originating from the trigeminal mesencephalic nucleus is unclear, and the possible innervation by parasympathetic nerve fibers has not been determined. In the present work, the retrograde axonal tracer, fast blue, was used to elucidate these questions and re-evaluated the literature data. The tracer was deposited in the supradiscal articular space of the rat TMJ, and an extensive morphometric analysis was performed of the labeled perikaryal profiles located in sensory and autonomic ganglia. This methodology permitted us to observe labeled small perikaryal profiles in the trigeminal ganglion, clustered mainly in the posterior-lateral region of the dorsal, medial and ventral thirds of horizontal sections, with some located in the anterior-lateral region of the ventral third. Sensory perikarya were also labeled in the dorsal root ganglia from C2 to C5. No labeled perikaryal profiles were found in the trigeminal mesencephalic nucleus. on the other hand, autonomic labeled perikaryal profiles were distributed in the sympathetic superior cervical and stellate ganglia, and parasympathetic otic ganglion. Our results confirmed those of previous studies and also demonstrated that: (i) there is a distribution pattern of labeled perikaryal profiles in the trigeminal ganglion; (ii) some perikaryal profiles located in the otic ganglion were labeled; and (iii) the trigeminal mesencephalic nucleus did not show any retrogradely labeled perikaryal profiles.