Modeling the Praia dos Ingleses submarine outfall, Florianópolis, Santa Catarina, Brazil.

An outfall 1,800 m long was initially proposed by CASAN (Water and Sanitation Company of Santa Catarina) to dispose of treated domestic residual water effluent from Praia dos Ingleses beach. The final plan average flow, estimated for the summer season is 0.1543 m³/s. The characteristics on the near field of the diffuser were calculated using the NRFIELD model supplied by a series of current profiles and density, gathered at the diffuser location between March and April, 2007. Reliable information of raw sewage fecal coliform concentration and nutrients has been supplied by CASAN, based on continuous monitoring of sewage in its treatment plants. The diffuser was designed in such a way that it maximizes the initial dilution allowing the nutrients concentration to comply with legislation limits. However, results showed that the treatment plant should be designed to mainly reduce total phosphorus. A hydrodynamic model was implemented to generate current fields for the far field modeling. Information gathered in the field on variation in water level, wind, and current velocity and direction was used for boundary and/or initial conditions. Then, a Lagrangian advective-diffusive model was used to evaluate the extent of the plumes. The results of far field modeling showed a need to increase the outfall length. A new location 3,300 m from the beach has been proposed.

Systolic dysfunction by two-dimensional speckle tracking echocardiography in dogs with parvoviral enteritis.

INTRODUCTION/OBJECTIVES: Parvoviral enteritis (PVE) can cause either primary or secondary myocardial injury; the latter is associated with systemic inflammatory response syndrome and sepsis. Strain (St) and strain rate (SR) are relatively new speckle tracking echocardiographic (STE) variables used to assess myocardial function and are less influenced by preload and volume status than are conventional variables. The aim of this study was to evaluate systolic function in dogs with PVE using two-dimensional STE. ANIMALS: Forty-five client-owned dogs were included. MATERIALS AND METHODS: Dogs were classified into four groups: healthy (n = 9), PVE-mild (n = 15), PVE-severe (n = 13) and PVE-died (n = 8). Left ventricular global and segmental myocardial St and SR were assessed in radial, circumferential and longitudinal axes in the right parasternal transverse and apical 4-chamber views. In the circumferential and longitudinal axes, the value of each segment was determined separately at the endocardial and epicardial levels. RESULTS: Compared to healthy animals, all dogs with PVE showed significantly impaired St and SR values, mainly for PVE-severe and PVE-died groups. Moreover, the lowest SR value was observed in the circumferential axis at the mid-septal epicardial segment in the PVE-died group. For this variable, a cut-off value of 0.95 s-1 demonstrated 100% sensitivity and specificity for distinguishing between PVE-severe and PVE-died groups. CONCLUSIONS: In the present study, all dogs with PVE developed systolic dysfunction, which was more severe in non-survivors. Assessment of St and SR in dogs with PVE might be clinically useful for evaluating haemodynamic status and developing suitable therapeutic strategies to improve prognosis.

Clostridium difficile toxins or infection induce upregulation of adenosine receptors and IL-6 with early pro-inflammatory and late anti-inflammatory pattern.

Clostridium difficile causes intestinal inflammation, which increases adenosine. We compared the expression of adenosine receptors (AR) subtypes A1, A2A, A2B, and A3 in HCT-8, IEC-6 cells, and isolated intestinal epithelial cells, challenged or not with Clostridium difficile toxin A and B (TcdA and TcdB) or infection (CDI). In HCT-8, TcdB induced an early A2BR expression at 6 h and a late A2AR expression at 6 and 24 h. In addition, both TcdA and TcdB increased IL-6 expression at all time-points (peak at 6 h) and PSB603, an A2BR antagonist, decreased IL-6 expression and production. In isolated cecum epithelial cells, TcdA induced an early expression of A2BR at 2s and 6 h, followed by a late expression of A2AR at 6 and 24 h and of A1R at 24 h. In CDI, A2AR and A2BR expressions were increased at day 3, but not at day 7. ARs play a role in regulating inflammation during CDI by inducing an early pro-inflammatory and a late anti-inflammatory response. The timing of interventions with AR antagonist or agonists may be of relevance in treatment of CDI.

Pharmacological properties of a pore induced by raising intracellular Ca2+.

Recent studies on the P2X(7) receptor in 2BH4 cells and peritoneal macrophages have demonstrated that the raise in intracellular Ca(2+) concentration induces a pore opening similar to P2X(7) receptor pore. Herein, we have investigated whether the pore activated by the elevation of intracellular Ca(2+) concentration is associated to P2X(7) receptor. Using patch clamp in cell attached, whole cell configuration, and dye uptake, we measured the pore opening in cell types that express the P2X(7) receptor (2BH4 cells and peritoneal macrophages) and in cells that do not express this receptor (HEK-293 and IT45-RI cells). In 2BH4 cells, the stimulation with ionomycin (5-10 microM) increased intracellular free Ca(2+) concentration and induced pore formation with conductance of 421 +/- 14 pS, half-time (t(1/2)) for ethidium bromide uptake of 118 +/- 17 s, and t(1/2) for Lucifer yellow of 122 +/- 11 s. P2X(7) receptor antagonists did not block these effects. Stimulation of HEK-293 and IT45-RI cells resulted in pore formation with properties similar to those found for 2BH4 cells. Connexin hemichannel inhibitors (carbenoxolone and heptanol) also did not inhibit the pore-induced effect following the increase in intracellular Ca(2+) concentration. However, 5-(N,N-hexamethylene)-amiloride, a P2X(7) receptor pore blocker, inhibited the induced pore. Moreover, intracellular signaling modulators, such as calmodulin, phospholipase C, mitogen-activated protein kinase, and cytoskeleton components were important for the pore formation. Additionally, we confirmed the results obtained for electrophysiology by using the flow cytometry, and we discarded the possibility of cellular death induced by raising intracellular Ca(2+) at the doses used by using lactate dehydrogenase release assay. In conclusion, increased concentration in intracellular Ca(+2) induces a novel membrane pore pharmacologically different from the P2X(7) associated pore and hemigap-junction pore.

A cation non-selective channel induced by extracellular ATP in macrophages and phagocytic cells of the thymic reticulum.

Extracellular ATP4- can bind to P2Z purinergic receptors including depolarization and cytoplasmic membrane permeabilization to small molecular weight solutes in macrophages, thymocytes, mast cells, phagocytic cells of the thymic reticulum and other cell types. An ATP(4-)-induced cation current has been described in whole-cell records of some of these cells but it is currently not clear whether these currents and the phenomenon of membrane permeabilization are a consequence of only one type of P2Z-associated channel/pore or two different phenomena triggered by one or more receptors. Here we use the outside-out patch-clamp technique to describe a single channel associated with this cation current in two murine phagocytic cells: intraperitoneal macrophages and phagocytic cells of the thymic reticulum. Multi channel currents could be readily observed in 77% of the outside-out patches of macrophages. Single channels of 7.8 pS could usually be resolved only in tail currents. Reversal potential measurements and ion replacement experiments indicated a lack of cation selectivity, similarly to what has already been described for the ATP(4-)-induced whole-cell inward current. No large-conductance channels that could explain the permeabilization to small molecular weight studies solutes was observed under our experimental conditions. A single channel of approx. 5 pS was also observed in phagocytic cells of the thymic reticulum under similar conditions. We conclude that the channel here described is the main carrier of cation current usually associated with the binding of ATP4- to P2Z receptors in whole-cell and outside-out patch-clamp experiments.

Characterization of P2Z purinergic receptors on phagocytic cells of the thymic reticulum in culture.

The thymic microenvironment is under intrinsic and extrinsic control circuits by several elements including hormones, neuropeptides, lymphokines, innervation and cell contact. P2 purinergic receptors have been described in a number of cells including macrophages, thymocytes, and other cells of the immune-inflammatory system. Here, we use the whole-cell patch-clamp technique and dye permeabilization assays to investigate the presence of ionic channels and purinergic receptors in one microenvironmental thymic component, namely the phagocytic cell of the thymic reticulum. At holding potentials ranging from -30 to -60 mV, applications of extracellular ATP in the vicinity of the cell membrane induce a transient and fast-activating inward current followed in most cells by an outward current. The whole event lasts 5-20 s. The inward current has a reversal potential close to 0 mV and the outward current can be ascribed to a Ca2+ -dependent K+ conductance. Both currents are inhibited by Mg2+, suggesting that the phenomenon is mediated by ATP4-. ATP-gamma-S can also induce both inward and outward currents. Exposure of phagocytic cells of the thymic reticulum to 5 mM ATP for 10 min induced permeabilization to lucifer yellow but not to the larger dyes trypan blue and rhodamine-dextran, suggesting a molecular weight cut-off smaller than 900. These observations lead us to conclude that phagocytic cells of the thymic reticulum express P2Z purinergic receptors that can mobilize Ca2+, induce the opening of ionic channels and permeabilize the cell membrane.

Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice.

Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury. We investigated whether apoE deficiency reduces the healing effect of alanyl-glutamine (Ala-Gln) treatment, a recognized gut-trophic nutrient, during tissue recovery after 5-FU-induced intestinal mucositis. APOE-knockout (APOE-/-) and wild-type (APOE+/+) C57BL6J male and female mice (N=86) were given either Ala-Gln (100 mM) or phosphate buffered saline (PBS) by gavage 3 days before and 5 days after a 5-fluorouracil (5-FU) challenge (450 mg/kg, via intraperitoneal injection). Mouse body weight was monitored daily. The 5-FU cytotoxic effect was evaluated by leukometry. Intestinal villus height, villus/crypt ratio, and villin expression were monitored to assess recovery of the intestinal absorptive surface area. Crypt length, mitotic, apoptotic, and necrotic crypt indexes, and quantitative real-time PCR for insulin-like growth factor-1 (IGF-1) and B-cell lymphoma 2 (Bcl-2) intestinal mRNA transcripts were used to evaluate intestinal epithelial cell turnover. 5-FU challenge caused significant weight loss and leukopenia (P<0.001) in both mouse strains, which was not improved by Ala-Gln. Villus blunting, crypt hyperplasia, and reduced villus/crypt ratio (P<0.05) were found in all 5-FU-challenged mice but not in PBS controls. Ala-Gln improved villus/crypt ratio, crypt length and mitotic index in all challenged mice, compared with PBS controls. Ala-Gln improved villus height only in APOE-/- mice. Crypt cell apoptosis and necrotic scores were increased in all mice challenged by 5-FU, compared with untreated controls. Those scores were significantly lower in Ala-Gln-treated APOE+/+ mice than in controls. Bcl-2 and IGF-1 mRNA transcripts were reduced only in the APOE-/- -challenged mice. Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.

Neuroendocrine control of the thymus.

Thymocytes undergo a complex process of differentiation, largely dependent on interactions with the thymic microenvironment, a tridimensional cellular network formed by epithelial cells, macrophages, dendritic cells, and fibroblasts. One key cellular interaction involves the TCR-CD3 complex expressed by thymocytes with MHC-peptide complexes present on microenvironmental cells. Additionally, thymic epithelial cells (TEC) interact with thymocytes via soluble polypeptides such as thymic hormones and interleukins, as well as through extracellular matrix (ECM) ligands and receptors. Such types of heterotypic interactions are under neuroendocrine control. For example, thymic endocrine function, represented by thymulin production, is up-regulated, both in vivo and in vitro, by thyroid and pituitary hormones, including prolactin and growth hormone. We also showed that these peptides enhance the expression of ECM ligands and receptors, as well as the degree of TEC-thymocyte adhesion. In addition, we studied the thymic nurse cell complex, used herein as an in vitro model for ECM-mediated intrathymic T-cell migration. We observed that T-cell migration is also hormonally regulated as ascertained by the thymocyte entrance into and exit from these lymphoepithelial complexes. Taken together these data clearly illustrate the concept that neuroendocrine circuits exert a pleiotropic control on thymus physiology. Lastly, the intrathymic production of classic hormones such as prolactin and growth hormone suggests that, in addition to endocrine circuits, paracrine and autocrine interactions mediated by these peptides and their respective receptors may exist in the thymus, thus influencing both lymphoid and microenvironmental compartments of the organ.

Gap junction modulation by extracellular signaling molecules: the thymus model.

Gap junctions are intercellular channels which connect adjacent cells and allow direct exchange of molecules of low molecular weight between them. Such a communication has been described as fundamental in many systems due to its importance in coordination, proliferation and differentiation. Recently, it has been shown that gap junctional intercellular communication (GJIC) can be modulated by several extracellular soluble factors such as classical hormones, neurotransmitters, interleukins, growth factors and some paracrine substances. Herein, we discuss some aspects of the general modulation of GJIC by extracellular messenger molecules and more particularly the regulation of such communication in the thymus gland. Additionally, we discuss recent data concerning the study of different neuropeptides and hormones in the modulation of GJIC in thymic epithelial cells. We also suggest that the thymus may be viewed as a model to study the modulation of gap junction communication by different extracellular messengers involved in non-classical circuits, since this organ is under bidirectional neuroimmunoendocrine control.

Oral lysine clonixinate in the acute treatment of migraine: a double-blind placebo-controlled study.

Several oral nonsteroidal anti-inflammatory drugs (NSAIDs) are effective to treat migraine attacks. Lysine clonixinate (LC) is a NSAID derived from nicotinic acid that has proven to be effective in various pain syndromes such as renal colic and muscular pain. The aim of this double-blind, placebo-controlled study was to evaluate the efficacy of oral LC compared to placebo in the acute treatment of migraine. Sixty four patients with the diagnosis of migraine, according to the IHS criteria, were studied prospectively. Patients received LC or placebo once the headache reached moderate or severe intensity for 6 consecutive attacks. With regard to the moderate attacks, LC was superior than placebo after 1, 2 and 4 hours. The consumption of other rescue medications after 4 hours was significantly higher in the placebo group. With regard to the severe attacks, there was no difference between the active drug group and the placebo group concerning headache intensity and consumption of other rescue medications. We conclude that the NSAID lysine clonixinate is effective in treating moderately severe migraine attacks. It is not superior than placebo in treating severe migraine attacks.

Is pannexin the pore associated with the P2X7 receptor?

The P2X7 receptor (P2X7R), an ATP-gated cation channel, is expressed predominantly in leukocytes. Activation of P2X7R has been implicated in the formation of a cytolytic pore (i.e., a large conductance channel) that allows the passage of molecules up to 900 Da in macrophages. At least two hypotheses have been presented to explain the conversion of a nonselective cation channel to a cytolytic pore. One hypothesis suggests that the pore is a separate molecular structure activated by P2X7R, and the second asserts that this is an intrinsic property of P2X7R (pore dilation). Based on connexin knockout and hemichannel antagonist studies, some groups have concluded that connexins and pannexins, the hemichannel-forming proteins in vertebrates, are fundamental components of the large conductance channel associated with P2X7R. Dye uptake and electrophysiology experiments were used to evaluate the efficacy and specificity of some hemichannel antagonists under conditions known to open the large conductance channel associated with P2X7R. Hemichannel antagonists and interference RNA (RNAi) targeting pannexin-1 did not affect P2X7R macroscopic currents [ATP, 1,570±189 pA; ATP+100 µM carbenoxolone (CBX), 1,498±100 pA; ATP+1 mM probenecid (Prob), 1,522±9 pA] or dye uptake in a FACS assay (ATP, 63±5%; ATP+100 µM CBX, 51.51±8.4%; ATP+1 mM Prob, 57.7±4.3%) in mouse macrophages. These findings strongly suggest that the high-permeability pore evident after prolonged P2X7R activation does not occur through connexin or pannexin hemichannels in murine macrophages. Another membrane protein may be involved in P2X7R pore formation.

Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice

Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury. We investigated whether apoE deficiency reduces the healing effect of alanyl-glutamine (Ala-Gln) treatment, a recognized gut-trophic nutrient, during tissue recovery after 5-FU-induced intestinal mucositis. APOE-knockout (APOE-/-) and wild-type (APOE+/+) C57BL6J male and female mice (N=86) were given either Ala-Gln (100 mM) or phosphate buffered saline (PBS) by gavage 3 days before and 5 days after a 5-fluorouracil (5-FU) challenge (450 mg/kg, via intraperitoneal injection). Mouse body weight was monitored daily. The 5-FU cytotoxic effect was evaluated by leukometry. Intestinal villus height, villus/crypt ratio, and villin expression were monitored to assess recovery of the intestinal absorptive surface area. Crypt length, mitotic, apoptotic, and necrotic crypt indexes, and quantitative real-time PCR for insulin-like growth factor-1 (IGF-1) and B-cell lymphoma 2 (Bcl-2) intestinal mRNA transcripts were used to evaluate intestinal epithelial cell turnover. 5-FU challenge caused significant weight loss and leukopenia (P<0.001) in both mouse strains, which was not improved by Ala-Gln. Villus blunting, crypt hyperplasia, and reduced villus/crypt ratio (P<0.05) were found in all 5-FU-challenged mice but not in PBS controls. Ala-Gln improved villus/crypt ratio, crypt length and mitotic index in all challenged mice, compared with PBS controls. Ala-Gln improved villus height only in APOE-/- mice. Crypt cell apoptosis and necrotic scores were increased in all mice challenged by 5-FU, compared with untreated controls. Those scores were significantly lower in Ala-Gln-treated APOE+/+ mice than in controls. Bcl-2 and IGF-1 mRNA transcripts were reduced only in the APOE-/--challenged mice. Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.

Determination of diflubenzuron residues in milk and cattle tissues / Determinação de resíduos de diflubenzuron em leite e tecidos bovinos

Diflubenzuron (DFB) is used to control ectoparasitic infestation by inhibiting larvae development in the manure and feces of treated animals. It is also currently been used to control tick infestations. In this study, milk and tissues from cattle treated orally with DFB for a 77-120 day period with a commercial product containing the compound were analyzed for the presence of residues. DFB residues were determined by using extraction with acetonitrile, cleanup with C18 SPE and chromatographic analysis by HPLC with UV detection (254nm). DFB was not detected in any of the analysed samples (<0.006mg kg-1 for fat, <0.014mg kg-1 for muscle, <0.015mg kg-1 for kidney, <0.016mg kg-1 for liver and <0.0006mg kg-1 for milk). In this manner, the use of this compound, according to the manufacturer's suggested doses may result in cattle milk, liver, kidneys, fat and muscles being considered safe regarding the presence of DFB residues.

O diflubenzuron (DFB) é um inibidor de desenvolvimento de insetos que inibe a síntese de quitina com atividade ovicida e larvicida e está sendo utilizado na pecuária para o controle do carrapato. Leite e tecidos provenientes de bovinos tratados por um período de 77 a 120 dias com um produto comercial contendo DFB foram analisados quanto à presença de resíduos. Os resíduos de DFB foram determinados utilizando-se extração com acetonitrila, limpeza por SPE C18 e cromatografia líquida de alta eficiência com detecção por UV (254nm). DFB não foi detectado em nenhuma das amostras analisadas (<0.006mg kg-1 para gordura, <0,014mg kg-1 para músculo, <0,015mg kg-1 para rim, <0,016mg kg-1 para fígado e <0.0006mg kg-1 para leite). Dessa forma, a utilização do princípio ativo conforme recomendado pelo fabricante e em níveis suficientes para se obter o efeito larvicida desejado deve resultar em leite, fígado, rins, gordura e músculos que podem ser considerados seguros para o consumo em termos da presença DFB.

Clinical applicability of natural product(s)-containing mouthwashes as adjunctive treatment of biofilm-induced gingivitis: a systematic review / Aplicabilidade clínica de colutórios à base de produtos naturais como tratamento adjuvante da gengivite induzida por biofilme: uma revisão sistemática

Natural products have emerged as an effective and low-cost alternative for treating various diseases of the oral cavity. This study aimed to evaluate, through a systematic literature review, if there is scientific evidence ensuring the safe and effective use of natural product(s)-containing mouthwashes as adjunctive treatment of biofilm-induced gingivitis. Searches were conducted in the databases Medline, SciELO, LILACS and Cochrane Library, by using combinations of the key words gingivitis/natural products/phytotherapy/mouthwash, in English, Portuguese and Spanish. Studies published until September 2010 were considered. Four examiners analyzed independently: study design and phase, methodological quality (Jadad scale - JE), experimental product and its concentration, dosing interval and time of usage, as well as employed statistical analysis and clinical outcome of interest. From the 503 articles found, 08 were included in the final review as phase II, controlled, randomized and blind clinical trials, scoring 4 (25%) and 5 (75%) in JE. The main natural products assessed were: Azadirachta indica, Garcinia mangostana, Lippia sidoides, Salvadora persica and Sesamum indicum whose concentration, dosing interval, time of usage and adverse effects varied according to each study. The Plaque and Gingival Index were most employed, as well as α = 5% and paired t, Student's t, Wilcoxon and Mann-Whitney tests. A total of 62.5% and 50% of the products significantly reduced supragingival biofilm and gingivitis, respectively. Mouthwashes containing the essential oil from the leaves of L. sidoides (1%) and the extract from the leaves of A. indica (25%) can be indicated as adjunctive treatment of biofilm-induced gingivitis.

Os produtos naturais têm surgido como alternativa eficaz e de baixo custo para o tratamento de várias doenças da cavidade oral. Objetivou-se avaliar, a partir de revisão sistemática da literatura, se há evidências científicas garantindo a utilização segura e eficaz de antissépticos bucais contendo produto(s) natural(is) como tratamento adjuvante da gengivite induzida por biofilme. Foram realizadas buscas nas bases de dados Medline, SciELO, LILACS e Cochrane Library, através de combinações usando as palavras-chave gengivite/produtos naturais/ fitoterápicos/bochechos, em Inglês, Português e Espanhol. Consideraram-se os estudos publicados até setembro de 2010. Quatro examinadores analisaram separadamente: desenho e fase do estudo, qualidade metodológica (escala de Jadad - EJ), produto experimental e a concentração, intervalo de administração e tempo de uso, bem como a análise estatística empregada e os resultados clínicos de interesse. Foram encontrados 503 artigos dos quais 08 foram incluídos na revisão final como sendo ensaios clínicos fase II, controlados, randomizados e cegos, marcando 4 (25%) e 5 (75%) na EJ. Os principais produtos naturais avaliados foram Azadirachta indica, Garcinia mangostana, Lippia sidoides, Salvadora persica e Sesamum indicum, cuja concentração, intervalo de administração, tempo de uso, e efeitos adversos, variaram de acordo com cada estudo. Índice de placa e Índice Gengival foram os mais utilizados, bem como α=5% e testes t-pareado, t-Student, Wilcoxon e Mann-Whitney. 62,5% e 50% dos produtos reduziram significativamente a presença de bioflme supragengival e gengivite, respectivamente. Os colutórios contendo o óleo essencial das folhas de L. sidoides (1%) e o extrato das folhas de A. indica (25%) podem ser indicados como tratamento adjuvante da gengivite induzida por biofilme.

Produção de antígeno e separação da proteína p28 por microfiltragem seriada para sorodiagnóstico da artrite encefalite caprina por ensaio imunoenzimático / Production of antigen and p28 protein separation by microfiltration serial for serodiagnosis of caprine arthritis encephalitis by enzyme immunoassay

Este estudo teve como objetivo produzir um antígeno (Ag) a partir de cultura de células de membrana sinovial caprina (MSC) infectadas com o vírus de artrite encefalite caprina (CAEV), pela técnica de microfiltração seriada, substituindo a ultracentrifugação em colchão de sacarose (UCCS) para utilização em ELISA indireto (ELISA-i). Amostras de 188 soros caprinos, que previamente foram testados pelo Western blot (WB) com Ag UCCS, foram submetidas à análise pelo ELISA-i com o novo antígeno produzido, que mostrou concordância de 92% em relação ao antígeno UCCS. A sensibilidade e a especificidade do ELISA em relação ao WB foram de 95,6% e 88,5%, respectivamente. A nova técnica, criada a partir de microfiltrações, mostrou-se efetiva e de baixo custo para o diagnóstico sorológico de anticorpos para CAEV em comparação ao antígeno ultracentrifugado, e constitui uma alternativa viável para produção de antígeno purificado de lentivírus de pequenos ruminantes.

This study aimed to produce an antigen (Ag) from the culture of goat synovial membrane cells (MSC) infected by CAEV through serial microfiltering technique replacing ultra ultracentrifugation in sacarosis Mattress (UCCS) for the indirect diagnosis ELISA tests (i ELISA). Samples of 188 sera from goats previously examined by Western Blot (WB) with Ag UCCS were submitted to analysis by i ELISA with new antigen produced, demonstrating an accordance of 92% in relation to UCCS antigen. The specificity and sensitivity relating to WB were of 95,65% and 88, 5% respectively. The new technique created from the microfiltering is effective and with low cost for the serological antibodies diagnosis of CAEV comparing to the ultracentrifuged one, presenting, therefore, as a viable alternative for purified antigen of lentivirus in small ruminants.

Alterações histopatológicas da glândula mamária e qualidade do leite de cabras naturalmente infectadas com o CAEV / Histopathological changes in the mammary gland and milk quality of goats naturally infected with CAEV

Avaliou-se a influência do vírus da CAE nas características físico-químicas de amostras de leite de 54 cabras, sem predileção racial, distribuindo-as em dois grupos: cabras positivas e negativas para o teste de imunodifusão em gel de agarose. As amostras de leite foram submetidas à análise ultrassônica para obtenção de parâmetros físico-químicos - gordura, extrato seco, proteínas, lactose e densidade; realização de microbiologia - bactérias mesófilas (UCF/mL). Foram coletadas amostras de tecido mamário para exame histopatológico e imunohistoquímica. Não houve diferença significativa das características avaliadas entre os dois grupos; no microbiológico, não houve relação direta da presença de mesófilas associada à infecção pelo CAEV. Na histopatologia, observaram-se áreas com infiltração celular de monócitos, polimorfonucleares, plasmócitos, fibrose, ausência de morfologia normal do parênquima mamário, denotando processo inflamatório crônico; e foi confirmada a presença do vírus na glândula pela imunohistoquímica. Os resultados não mostraram relação direta da incidência da CAE como fator negativo no desenvolvimento do rebanho.

Aiming to evaluate the influence of CAE viruses in the chemical and physical characteristics of milk, the samples were collected from 54 goats, without racial predilection, these were divided into two groups: goats positive and negative according results of test Agarose Gel Immunodiffusion. Milk samples were ultrasonic analyzed to obtain physicochemical parameters (fat, solids, protein, lactose and density); performance microbiology (mesophilic bacteria - CFU/mL) and mammary gland samples were collected for evaluation histopathology and immunohistochemistry. The results of physical-chemical analysis showed no significant difference between the milk samples of two groups. In the microbiological analysis showed the presence of aerobic mesophilic bacteria, but this change is not associated with the presence of CAEV infection. On histopathology, there were areas with infiltration of mononuclear-leukocyte and polymorph nuclear, plasma cells, fibrosis and absence of normal morphology of the mammary tissue, indicating a chronic inflammatory process; and confirmed the presence of virus, in the gland, by immunohistochemistry. The results showed no direct relationship between incidence of CAE in the herd as a negative factor for its development, however it is known that the disease in its chronic nature, causes reduction in the productivity of the herd.

Prevalência de anticorpos contra Leptospira spp. em bovinos, caninos, equinos, ovinos e suínos do município de Jaguapitã, estado do Paraná, Brasil / Prevalence of antibodies against Leptospira spp. in cattle, dogs, horses, sheep and swine of Jaguapitã, state of Paraná, Brazil

O objetivo deste trabalho foi determinar a prevalência de anticorpos contra Leptospira spp. em bovinos, caninos, equinos, ovinos e suínos, oriundos de 40 propriedades localizadas na área rural do Município de Jaguapitã, Estado do Paraná. Foram colhidas amostras de sangue de 370 bovinos, 161 equinos, 70 ovinos, 230 suínos e 97 caninos. O número de animais testados em cada propriedade, assim como o número de propriedades, foi determinado utilizando-se o programa Epi-info versão 6. As amostras de soros obtidas foram submetidas à prova de soroaglutinação microscópica (SAM) com 22 sorovares de Leptospira spp. Das 40 propriedades rurais pesquisadas, 38 (95,00%) tiveram pelo menos um animal sororeagente na SAM e dos 928 animais estudados, 316 (34,08%) apresentaram títulos ? 100. A prevalência observada na espécie bovina foi de 42,43%, com 87,18% das propriedades apresentando pelo menos um animal sororeagente. As prevalências de animais e propriedades reagentes para as demais espécies foram, respectivamente, 48,44% e 87,18% para equinos; 38,57% e 100% para ovinos; 18,70% e 28,00% para suínos; 11,34% e 31,25% para cães. O sorovar mais provável encontrado em bovinos foi Hardjo, em equinos Castellonis e Sentot, em ovinos, suínos e cães Icterohaemorrhagiae. Os resultados obtidos neste trabalho demonstram que as cinco espécies animais estudadas na área rural do Município de Jaguapitã tiveram contato com vários sorovares de Leptospira spp. Além disso, os resultados sorológicos sugerem uma possível transmissão do micro-organismo entre espécies animais, provavelmente em decorrência da exposição às mesmas fontes de infecção entre os animais estudados.

The objective of this work was to determine the prevalence of antibodies against Leptospira spp. in cattle, dogs, horses, sheep and swine from 40 properties located in the rural area of Jaguapitã, state of Paraná, Brazil. Blood samples were taken from 370 cattle, 97 dogs, 161 horses, 70 sheep and 230 swine. The number of animals tested on each property, as well as the number of properties was determined using the program Epi-info version 6. Samples of serum were submitted to microscopic agglutination test (MAT) with 22 Leptospira spp. serovars. From the 40 rural properties investigated, 38 (95.00%) had at least one positive animal according to SAM, and from 928 studied animals, 316 (34.08%) presented titers ? 100. The prevalence observed in the bovine species was 42.43%, with 87.18% of the properties presenting at least one positive animal. The prevalence of animals and properties reactive for the other species were, respectively: 48.44% and 87.18% for horses; 38.57% and 100% for sheep; 18.70% and 28.00% for swine; 11.34% and 31.25% for dogs. The most frequent serovar in bovines was Hardjo, in horses Castellonis and Sentot, in sheep, swine and dogs Icterohaemorrhagiae. The results obtained in this study demonstrated that the 5 studied animal species in the rural area of Jaguapitã had contact with several Leptospira spp. serovars. Moreover, serological results suggest a possible transmission of Leptospira spp. between animal species, probably because of exposure to the same sources of infection among the animals studied.

Procedures to characterize and study P2Z/P2X7 purinoceptor: flow cytometry as a promising practical, reliable tool.

The expression of P2Z/P2X7 purinoceptor in different cell types is well established. This receptor is a member of the ionotropic P2X receptor family, which is composed by seven cloned receptor subtypes (P2X1 - P2X7). Interestingly, the P2Z/P2X7 has a unique feature of being linked to a non-selective pore which allows the passage of molecules up to 900 Da depending on the cell type. Early studies of P2Z/P2X7 purinoceptor were exclusively based on classical pharmacological studies but the recent tools of molecular biology have enriched the analysis of the receptor expression. The majority of assays and techniques chosen so far to study the expression of P2Z/P2X7 receptor explore directly or indirectly the effects of the opening of P2Z/P2X7 linked pore. In this review we describe the main techniques used to study the expression and functionality of P2Z/P2X7 receptor. Additionally, the increasing need and importance of a multifunctional analysis of P2Z/P2X7 expression based on flow cytometry technology is discussed, as well as the adoption of a more complete analysis of P2Z/P2X7 expression involving different techniques.