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This study was conducted to assess the effects of functional oil (FO) blend on performance, blood metabolites, organ biometry and intestinal morphometry in piglets. A total of 128 crossbreed piglets (Landrace × Large White, 64 uncastrated males and 64 females, 21 d of age, and 6.79 ± 1.76 kg BW) were allocated in a randomized complete block design with two dietary treatments: a FO-free (FOF) diet or a diet based on added FO (1,500 mg/kg of diet with castor oil plus cashew nutshell oil). Piglets fed FO showed higher (p ≤ 0.05) average daily feed intake, daily body weight gain and final body weight after 23 d of study. For the total period, piglets fed FO showed greater (p = 0.007) feed conversion ratio. On d 23, higher serum total protein (p = 0.026) and globulin (p = 0.050) concentration, lower liver (p = 0.042) and stomach (p = 0.074) weight, and greater (p = 0.082) villi height (VH) in duodenum were observed in piglets fed FO. Nonetheless, piglets fed FOF showed greater (p = 0.054) ileal VH, but greater (p = 0.004) crypt depth (CD) in jejunum. Piglets fed FO showed higher VH to CD ratio in jejunum (p = 0.068) and duodenum (p = 0.074) on d 23 and 37, respectively. Based on the results, FO blend improved the performance of weaned piglets; however, it negatively affected the feed conversion ratio in the total period. Moreover, FO blend promoted changes in total protein concentrations and improvements in digestive and absorptive capacity assessed through VH to CD ratio, with a significant reduction in organs.

Este estudo foi conduzido para avaliar os efeitos de uma mistura de óleo funcional (OF) no desempenho zootécnico, nos metabólitos sanguíneos, na biometria de órgãos e na morfometria intestinal de leitões. Um total de 128 leitões mestiços (Landrace × Large White, 64 machos inteiros e 64 fêmeas; 21 dias de idade e peso corporal de 6,79 ± 1,76 kg) foram alocados em um delineamento de blocos casualizados completos, com dois tratamentos dietéticos: uma dieta sem OF (SOF) ou uma dieta baseada na adição de OF (1.500 mg/kg de dieta com OF de mamona e castanha de caju). Os leitões alimentados com OF apresentaram maior (p ≤ 0,05) consumo de ração diário médio, ganho de peso corporal diário e peso corporal aos 23 dias de experimento. Entretanto, os leitões que consumiram dietas com OF tiveram conversão alimentar superior (p = 0,007) no período total. No dia 23, houve aumento na concentração de proteína total (p = 0,026) e globulina (p = 0,050), menor peso de fígado (p = 0,042) e estômago (p = 0,074), e maior (p = 0,082) altura de vilosidade (AV) no duodeno em leitões que consumiram OF; entretanto, os leitões SOF tiveram (p = 0,054) AV superior no íleo, mas apresentaram (p = 0,004) profundidade de cripta (PC) superior no jejuno. Uma maior relação AV:PC no jejuno (p = 0,068) e duodeno (p = 0,074) foi observada em leitões com OF nos dias 23 e 37, respectivamente. Com base nos resultados, a mistura de OF melhorou o desempenho dos leitões desmamados; no entanto, afetou negativamente a taxa de conversão alimentar no período total. Além disso, a mistura de OF promoveu alterações nas concentrações proteínas totais e melhorias na capacidade digestiva e absortiva avaliadas através da relação AV:PC, com uma redução significativa nos órgãos.

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Introduction. Biofilm formation is a major virulence factor associated with Staphylococcus aureus infections. However, the influence of plasma proteins on biofilm formation of clinical isolates in vitro remains unclear.Hypotheses. We hypothesized that coating surfaces with plasma proteins might induce biofilm formation by S. aureus of different clonal lineages.Aim. To evaluate biofilm production by clinical S. aureus isolates of different clonal lineages isolated in Rio de Janeiro hospitals and investigated the presence of biofilm-associated genes.Methodology. This study assessed biofilm production of 60 S. aureus isolates in polystyrene microtitre plates with and without fibrinogen or fibronectin. The biochemical composition of the biofilm matrices was determined and the biofilm formation on fibrinogen-coated surfaces was also evaluated by confocal laser scanning microscopy. The presence of biofilm-related genes was detected by PCR, and the typing and functionality of agr operon was also evaluated.Results. Most of the isolates (45 %) were weak biofilm producers or non-producers. However, most of them presented a significant increase in biofilm production on plates covered with plasma proteins. There was no significant difference in biofilm formation between methicillin-resistant and -susceptible S. aureus isolates, or between different clonal lineages, except for ST30-IV (weak producers) and ST239-III (strong producers). The fnbB gene was associated with higher biofilm production.Conclusion. An increase in biofilm production in the presence of plasma proteins highlights the importance of investigating biofilm formation by S. aureus clinical isolates under different conditions since this virulence factor contributes to persistent infections and increased resistance to antimicrobials.

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Precipitation of blood products from plasma fractionation has played a fundamental role in the industrial purification of important therapeutic products. Only a few studies have been reported by using tannins as proteins precipitant agent from whole plasma while, several conditions have been analyzed. Here, we decided to verify the effect of the temperature on the precipitation process of plasma proteins using tannic acid (TA). Plasma proteins were precipitated with tannic acid by using different temperature incubations. Subsequently, the protein-TA complex was analyzed by SDS-PAGE and quantified. In addition, the protein activity of the complex was measured after heating, as well as the structural changes of the complexes were accompanied by thermogravimetric analysis, differential scanning calorimetry and circular dichroism. In all conditions tested, tannic acid was able to precipitate without selectively separating the proteins in the mixture by using different temperatures during the precipitation process. Furthermore, the protein concentration from the plasma precipitate was not affected by different temperatures and the plasma precipitate was able to dissolve fibrin clots in vitro.

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Seminal plasma proteins already demonstrated to reflect the testicular environment function and important regulatory mechanisms. However, it is crucial to understand which of these proteins participate in probable altered pathways in testicular germ cell tumours and after unilateral orchiectomy. In this study, we proposed to verify, by a multiplex approach, the levels of DNA damage and apoptosis pathways' proteins, in seminal plasma of men before and after unilateral orchiectomy, and also in control men. Comparing pre- and post-orchiectomy groups, just the apoptosis pathways' proteins presented different levels, in which Bad was lower and Bcl2, Akt, caspase-9, p53 and caspase-8 were higher after orchiectomy. When comparing pre- and post-orchiectomy groups with control, both presented lower levels of ChK1, Chk2, H2AX, p53 and p21, for DNA damage pathway. Regarding the apoptosis pathway, lower levels of JNK, Bcl2, Akt, caspase-9, p53 and caspase-8 and higher levels of Bad were observed before orchiectomy. The post-orchiectomy group did not differ from controls, demonstrating a probable restoration on its proteins levels. We can conclude that testicular tumours can alter both of the assessed pathways, and its removal is associated with a probable restoration of the apoptosis pathway.

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Heparin was immobilized on magnetic chitosan particles to be used as a tool for human plasma protein identification. Chitosan was magnetized by co-precipitation with Fe2+/Fe3+ (MAG-CH). Heparin was functionalized with carbodiimide and N-hydroxysuccinimide and covalently linked to MAG-CH (MAG-CH-hep). X-ray diffraction confirmed the presence of chitosan and Fe3O4 in MAG-CH. This particle exhibited superparamagnetism and size between 100-300 µm. Human plasma diluted with 10 mM phosphate buffer (pH 5.5) or 50 mM Tris-HCl buffer (pH 8.5) was incubated with MAG-CH-hep, and the proteins fixed were eluted with the same buffers containing increasing concentrations of NaCl. The proteins obtained were investigated by SDS-PAGE, LC/MS, and biological activity tests (PT, aPTT, and enzymatic chromogenic assay). Inhibitors of the serpin family, prothrombin, and human albumin were identified in this study. Therefore, MAG-CH-hep can be used to purify these proteins and presents the following advantages: low-cost synthesis, magnetic separation, ion-exchange purification, and reusability.

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The study of wildlife health greatly contributes to understanding population dynamics and detecting conservation threats. The determination of the different fractions of plasma proteins (proteinogram) is an important laboratory tool to study wildlife health. The aim of this study was to characterize protein electrophoresis in wild Andean condors (Vultur gryphus) from north-western Patagonia and to evaluate differences according to age and sex classes. Once reference values of wild, apparently healthy individuals, were established, we compared these values to those of individuals received at the Buenos Aires Zoo in Argentina for rehabilitation due to various health problems. Reference proteinograms from wild Andean condors differed only in the α 1 and ß 2-fractions between sex categories. Males showed higher concentrations of these protein fractions than females. We found clear differences between wild birds and rehabilitating individuals. Total proteins, globulins, α 1-globulins, total α-globulins, ß 2-globulins, total ß-globulins, and γ-globulins were significantly higher in rehabilitating than in wild individuals, whereas albumin, α 2, and ß1-globulins were similar between these groups. The albumin/globulin ratio, as a general indicator of health, was significantly lower in rehabilitating than in wild individuals. The results indicate the effects on different protein fractions of pathologic processes occurring in individuals undergoing rehabilitation. Our results provide useful insights, contributing to improving diagnoses and prognoses in this species. This information may also be useful to assess the health status of Andean condors in studies of wild populations and for comparisons with other bird species.

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The search for plasma proteins precipitation methods has been increasing due to the plasma protein therapeutic needs in world-wide. Thus, this work evaluates the tannic acid (TA) ability to precipitate proteins from human plasma. In this study, TA-plasma protein complexes were studied at different pH conditions, tannin/plasma ratio and reaction mixing time. The complexes formed from combinations of TA and plasma proteins were analyzed by gel electrophoresis, protein quantification, particle size, charge, mass spectrometry, microscopic image, and circular dichroism. It was possible to verify the precipitate formation in all tested pH values, with high precipitation at pH 5. The native PAGE analysis showed three mainly bands corresponding independent of the pHs used. It was possible to observe a gradual growing of precipitate protein in the first precipitation process (P1) when increased the TA/plasma ratio. 15 min of incubation was enough to precipitate 72.3% of proteins. Spectroscopic analyzes showed albumin signals and the electron microscopy analysis of IgG-TA confirmed the compact form of a precipitate. According to CD, formation of the IgG-TA complexes does not cause a major structural change of the protein. From the results obtained, it was possible to establish some parameters for plasma proteins precipitation using TA.

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Leptospires are highly motile spirochetes equipped with strategies for efficient invasion and dissemination within the host. Our group previously demonstrated that pathogenic leptospires secrete proteases capable of cleaving and inactivating key molecules of the complement system, allowing these bacteria to circumvent host's innate immune defense mechanisms. Given the successful dissemination of leptospires during infection, we wondered if such proteases would target a broader range of host molecules. In the present study, the proteolytic activity of secreted leptospiral proteases against a panel of extracellular matrix (ECM) and plasma proteins was assessed. The culture supernatant of the virulent L. interrogans serovar Kennewicki strain Fromm (LPF) degraded human fibrinogen, plasma fibronectin, gelatin, and the proteoglycans decorin, biglycan, and lumican. Interestingly, human plasminogen was not cleaved by proteases present in the supernatants. Proteolytic activity was inhibited by 1,10-phenanthroline, suggesting the participation of metalloproteases. Moreover, production of proteases might be an important virulence determinant since culture-attenuated or saprophytic Leptospira did not display proteolytic activity against ECM or plasma components. Exoproteomic analysis allowed the identification of three metalloproteases that could be involved in the degradation of host components. The ability to cleave conjunctive tissue molecules and coagulation cascade proteins may certainly contribute to invasion and tissue destruction observed upon infection with Leptospira.

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Leptospires are highly motile spirochetes equipped with strategies for efficient invasion and dissemination within the host. Our group previously demonstrated that pathogenic leptospires secrete proteases capable of cleaving and inactivating key molecules of the complement system, allowing these bacteria to circumvent host's innate immune defense mechanisms. Given the successful dissemination of leptospires during infection, we wondered if such proteases would target a broader range of host molecules. In the present study, the proteolytic activity of secreted leptospiral proteases against a panel of extracellular matrix (ECM) and plasma proteins was assessed. The culture supernatant of the virulent L. interrogans serovar Kennewicki strain Fromm (LPF) degraded human fibrinogen, plasma fibronectin, gelatin, and the proteoglycans decorin, biglycan, and lumican. Interestingly, human plasminogen was not cleaved by proteases present in the supernatants. Proteolytic activity was inhibited by 1,10-phenanthroline, suggesting the participation of metalloproteases. Moreover, production of proteases might be an important virulence determinant since culture-attenuated or saprophytic Leptospira did not display proteolytic activity against ECM or plasma components. Exoproteomic analysis allowed the identification of three metalloproteases that could be involved in the degradation of host components. The ability to cleave conjunctive tissue molecules and coagulation cascade proteins may certainly contribute to invasion and tissue destruction observed upon infection with Leptospira.

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Optional function of body systems depends upon fluid and electrolyte balance; however, across the lifespan, disorders of fluid and electrolytes offset this, and the causative factors are varied. Nurses play a major role in the management of fluid and electrolyte balance. This article focuses on the role total body water content, plasma proteins, kidney function, and drug metabolism have on the age-related physiology impacting fluid and electrolyte balance, and on nursing implications.

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The aim of this work was to improve the quality of gluten-free bread, incorporating plasma bovine proteins concentrated by ultrafiltration and freeze-dried with saccharides (inulin and sucrose). The influence of these compounds on textural properties and final bread quality was assessed. The textural studies revealed that with the addition of proteins and inulin, homogeneous and smaller air cells were achieved improving the textural properties while the bread hardness was comparable with breads with gluten. The volume of gluten-free breads increased with increasing proteins and inulin concentrations, reaching a maximum at a protein concentration of 3.5% (w/w). The addition of the enhancers improved moisture retention of the loaves after cooking and an increase of lightness of crumb with respect to the control was observed. The sensory analysis found no statistically significant difference in sensory attributes evaluated with respect to the control, so these ingredients do not negatively affect the organoleptic properties of bread.

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Background: Seminal plasma is a set of secretions from accessory male reproduction organs which infl uence spermaticfunctions. Climate changes that affect gametogenesis may produce low reproduction effi ciency in bulls. Electrophoresisis a great help in the diagnosis of reproduction pathologies and in animal differentiation with regard to fertility within thecontext of climate changes. Current research aims at studying the infl uence of the rainy and dry periods in testicle and epididymis morphometry, semen characteristics and protein of seminal plasma by SDS-PAGE electrophoresis in extensivelybred Nellore and Simental bulls.Materials, Methods & Results: Five Nellore and two Simental bulls, at mean age of four years, were employed. Semencollection was undertaken during the rainy (spring and summer) and dry (autumn and winter) at 15-day intervals. Onehundred and fi fty-four ejaculations were analyzed and scrotal, testicle and epididymis measurement was provided. Samplesof semen plasma were centrifuged and conditioned at -196°C till electrophoresis processing. Proteins were extracted from200 µL from each sample in an extraction buffer composed of 0.625 M Tris - HCl, pH 6.8; 2% SDS, 5% β - mercaptoethanol and 20% glycerol. Proteins were quantifi ed and electrophoreses processed. Data were evaluated by analysis ofvariance and means compared by Tukey´s test at 5%. The expression TV = 0.0396 x (average testicle length) x (scrotalperimeter)2 was used for testicle volume (TV), with TV = 460.14 cm3 for the Simental breed during the rainy season andTV = 571.26 cm3 during the dry season. Nellore bulls showed TV = 524.75 cm3 during the rainy season and 515.13 cm3during the dry season. TV increased in Simental bulls during the dry period, whereas Nellore breed increased during therainy one. There was an increase (P < 0.05) in big and total defects during the rainy season...

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Background: Seminal plasma is a set of secretions from accessory male reproduction organs which infl uence spermaticfunctions. Climate changes that affect gametogenesis may produce low reproduction effi ciency in bulls. Electrophoresisis a great help in the diagnosis of reproduction pathologies and in animal differentiation with regard to fertility within thecontext of climate changes. Current research aims at studying the infl uence of the rainy and dry periods in testicle and epididymis morphometry, semen characteristics and protein of seminal plasma by SDS-PAGE electrophoresis in extensivelybred Nellore and Simental bulls.Materials, Methods & Results: Five Nellore and two Simental bulls, at mean age of four years, were employed. Semencollection was undertaken during the rainy (spring and summer) and dry (autumn and winter) at 15-day intervals. Onehundred and fi fty-four ejaculations were analyzed and scrotal, testicle and epididymis measurement was provided. Samplesof semen plasma were centrifuged and conditioned at -196°C till electrophoresis processing. Proteins were extracted from200 µL from each sample in an extraction buffer composed of 0.625 M Tris - HCl, pH 6.8; 2% SDS, 5% β - mercaptoethanol and 20% glycerol. Proteins were quantifi ed and electrophoreses processed. Data were evaluated by analysis ofvariance and means compared by Tukey´s test at 5%. The expression TV = 0.0396 x (average testicle length) x (scrotalperimeter)2 was used for testicle volume (TV), with TV = 460.14 cm3 for the Simental breed during the rainy season andTV = 571.26 cm3 during the dry season. Nellore bulls showed TV = 524.75 cm3 during the rainy season and 515.13 cm3during the dry season. TV increased in Simental bulls during the dry period, whereas Nellore breed increased during therainy one. There was an increase (P < 0.05) in big and total defects during the rainy season...(AU)

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This work deals with the effect of the addition of inulin and bovine plasma proteins as fat replacers, on the quality of minced meat. The proteins are obtained by ultrafiltration and freeze-drying. The following determinations were carried out: chemical composition, sensorial analysis (color, flavor, taste and consistency), emulsion stability and instrumental texture analysis of samples. The resulting formulations were compared with full-fat minced meat, as control. The results showed an increase of protein contents after fat replacement, while a fat reduction of 20-35% produced light products enriched with proteins and inulin as the functional ingredient. No change was observed in color, flavor, or taste among the samples. However, the sensory analysis showed that the combination of plasma protein (2.5%w/w) and inulin (2%w/w) had the best acceptability with respect to consistency, and had a lower fat drain from the emulsion. Texture profile analysis revealed that this formulation assimilated the control texture properties, being that this result is required for adequate consumer acceptance.

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Plants have emerged as an attractive alternative to the traditional mammalian cell cultures or microbial cell-based systems system for the production of valuable recombinant proteins. Through recombinant DNA technology, plants can be engineered to produce large quantities of pharmaceuticals and industrial proteins of high quality at low costs. The recombinant production, by transgenic plants, of therapeutic proteins normally present in human plasma, such as cytokines, coagulation factors, anticoagulants, and immunoglobulins, represents a response to the ongoing challenges in meeting the demand for therapeutic proteins to treat serious inherited or acquired bleeding and immunological diseases. As the clinical utilization of fractionated plasma molecules is limited by high production costs, using recombinant biopharmaceuticals derived from plants represents a feasible alternative to provide efficient treatment. Plant-derived pharmaceuticals also reduce the potential risks to patients of infection with pathogens or unwanted immune responses due to immunogenic antigens. In this review, we summarize the recent advances in molecular farming of cytokines. We also examine the technological basis, upcoming challenges, and perspectives for the biosynthesis and detection of these molecules in different plant production platforms.

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As parasitoses gastrintestinais constituem o principal entrave sanitário à ovinocaprinocultura, principalmente devido à resistência aos anti-helmínticos. Uma das formas de limitar a progressão da resistência aos fármacos é evitar a sua utilização em lotes ou rebanhos inteiros, restringindo seu uso aos animais que necessitam tratamento (tratamento seletivo). O objetivo deste trabalho foi estudar o uso dos escores corporal (EC) e de diarreia (ED) como indicadores do tratamento seletivo em ovinos, em comparação com o método FAMACHA© (GF) e indicadores laboratoriais de parasitismo. Foram acompanhadas quinzenalmente 15 ovelhas em lactação (agosto-novembro 2009) e após o desmame (dezembro 2009 ­ fevereiro de 2010) pelo GF, EC, ED, e análises laboratoriais para determinação do hematócrito (HT), proteínas plasmáticas totais (PPT) e contagem de ovos de helmintos por grama de fezes (OPG). A média de OPG se manteve estável, exceto no segundo mês de lactação (média 6057,69 OPG). Durante todo o acompanhamento, o GF foi moderadamente correlacionado ao OPG (0,37); EC (-0,32); PPT (-0,34) e HT (-0,54). Para o EC, os valores de correlação com o OPG foram de -0,35; -0,32 com GF; 0,45 com PPT e de 0,50 com HT. O ED não se mostrou um bom indicador indireto de verminose, pois não foi obtida nenhuma correlação significativa com as outras variáveis estudadas. Como o escore corporal foi correlacionado às variáveis laboratoriais em índices semelhantes aos obtidos pelo método FAMACHA© , seu uso é sugerido como ferramenta adicional de tratamento seletivo para ovelhas em lactação e nos meses subsequentes ao desmame.

The gastrointestinal parasitic infections constitute the main obstacle to the health of sheep and goat, mainly due to anthelmintic resistance. An alternative to limit the progression of drug resistance is to avoid to deworm the hole heard or group, treating individually only those animals that require so (selective treatment). The goal of this work was to study the condition score (BS) and the diarrhea score (DS) as indicators of selective treatment in sheep, in comparison to the FAMACHA© system (EF) and to laboratory analyses that indicate parasitism. Fifteen lactating (Aug.- Nov. 2009) or post weaning (Dez. 2009- Feb.2010) ewes were sampled fortnightly for EF, BS, DS and also for the laboratorial analysis to determine packed cell value (PCV/HT), total plasma protein fraction (TPP) and faecal egg counts (FEC/OPG). The mean FEC remained stable except in the second month of lactation period (average 6057.69 EPG). Throughout the monitoring, the GF was moderately correlated with FEC (0.37), BS (-0.32), TPP (-0.34) and PCV (-0.54). For the BS, the value of correlation with FEC was -0.35, -0.32 to GF; 0.45 to TPP and 0.50 to PCV. The DS was not a good indicator of parasite infection, because there was no significant correlation obtained with the other variables. As the body condition score was correlated to the laboratory variables in similar indices of the FAMACHA© system, we suggest the its use as an additional tool of selective treatment for ewes during lactation period and after weaning.

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The present study was designed to investigate the topographical distribution of seminal plasma (SP) proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS) and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.(AU)

Investigou-se a distribuição topográfica da ligação de proteínas seminais à membrana de espermatozoides bovinos epididimários e ejaculados. Utilizando imunocitoquímica e microscopia confocal, avaliaram-se a topografia de ligação das proteínas BSP-A3, albumina, transferrina, prostaglandina D sintetase (PGDS) e nucleobindina 2 (NUC2) à membrana espermática. Os experimentos foram realizados utilizando espermatozoides de cinco touros. Os resultados mostraram que, para espermatozoides epididimários, somente detectou-se a PGDS na crista do acrossomo. Nos espermatozoides ejaculados, a PGDS ligou-se de forma mais intensa à crista acrossômica, enquanto a NUC2 apresentou sinal bastante fraco na peça intermediária e região equatorial. A BSP-A3 ligou-se a todas as regiões estudadas, de forma mais intensa na peça intermediária e acrossomo. Nenhum sinal foi detectado para albumina ou transferrina, seja em espermatozoides epididimários ou ejaculados. Concluiu-se que PGDS, BSP-A3 e NUC2 interagem diretamente com espermatozoides bovinos, e mostrou distribuição topográfica específica. Estes achados permitem melhor compreensão sobre o papel desempenhado por essas proteínas na regulação da função espermática e da fertilidade.(AU)

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The present study was designed to investigate the topographical distribution of seminal plasma (SP) proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS) and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.

Investigou-se a distribuição topográfica da ligação de proteínas seminais à membrana de espermatozoides bovinos epididimários e ejaculados. Utilizando imunocitoquímica e microscopia confocal, avaliaram-se a topografia de ligação das proteínas BSP-A3, albumina, transferrina, prostaglandina D sintetase (PGDS) e nucleobindina 2 (NUC2) à membrana espermática. Os experimentos foram realizados utilizando espermatozoides de cinco touros. Os resultados mostraram que, para espermatozoides epididimários, somente detectou-se a PGDS na crista do acrossomo. Nos espermatozoides ejaculados, a PGDS ligou-se de forma mais intensa à crista acrossômica, enquanto a NUC2 apresentou sinal bastante fraco na peça intermediária e região equatorial. A BSP-A3 ligou-se a todas as regiões estudadas, de forma mais intensa na peça intermediária e acrossomo. Nenhum sinal foi detectado para albumina ou transferrina, seja em espermatozoides epididimários ou ejaculados. Concluiu-se que PGDS, BSP-A3 e NUC2 interagem diretamente com espermatozoides bovinos, e mostrou distribuição topográfica específica. Estes achados permitem melhor compreensão sobre o papel desempenhado por essas proteínas na regulação da função espermática e da fertilidade.

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RESUMO

No presente protocolo experimental, determinaram-se os proteinogramas séricos, por intermédio da eletroforese em gel de poliacrilamida contendo duodecil sulfato de sódio (SDS-PAGE), de 120 cães com raças e idades variadas e atendidos junto ao Hospital Veterinário "Governador Laudo Natel" da FCAV/Unesp, com o objetivo principal de comparar diferentes frações seroproteicas em estados anêmicos regenerativos, arregenerativos, imunomediados primários e secundários. Os referidos animais foram distribuídos em cinco grupos experimentais: grupo 1: 20 cães de controle; grupo 2: 28 cães com anemia regenerativa não imune; grupo 3: 27 cães com anemia arregenerativa não imune; grupo 4: 10 cães com anemia hemolítica imunomediada primária; grupo 5: 35 cães com anemia hemolítica imunomediada secundária. A técnica SDS-PAGE permitiu o fracionamento de 24 proteínas, cujos pesos moleculares (PM) variaram de 18.000 a 165.000 daltons (Da). Os cães com AHIM primária e secundária apresentaram 24 frações proteicas em seus traçados eletroforéticos, enquanto que cães de controle (1) e portadores de anemia regenerativa (2) e arregenerativa (3) de natureza não imune apresentaram 23 frações de proteínas, cuja proteína de peso molecular 68.000Da não foi encontrada. Dessa forma, 23 frações proteicas foram detectadas e revelaram-se comuns aos proteinogramas dos cães de controle e daqueles dos quatro grupos experimentais. Destas, identificaram-se nominalmente 11 frações proteicas, e as demais foram estudadas com base nos seus respectivos pesos moleculares. Em relação aos cães de controle, os anêmicos (grupos 2, 3, 4 e 5) apresentaram maiores concentrações de transferrina sérica e entre estes os animais portadores da AHIM primária. Todos os cães anêmicos apresentaram teores séricos de haptoglobina e fosforilase significativamente maiores que os controles, enquanto que a concentração sérica de ceruloplasmina foi significativamente maior nestes. Tais achados analisados em...

This assay aimed to determine the serum protein - via polyacrylamide gel electrophoresis, which contained duodecil sodium sulfate (SDS-PAGE) - in 120 dogs, with different breeds and ages, seen by the Veterinary Hospital "Governor Laudo Natel." These animals were grouped into five experimental groups: Group 1 - group control with 20 dogs, group 2 - 28 dogs with regenerative anemia; group 3 - 27 dogs with arregenarative; anemia group 4 - 10 dogs with primary immune-mediated hemolytic anemia (AHIM 1.rd); group 5 - 35 dogs with secondary immune-mediated hemolytic anemia (AHIM 2.rd). The technique allowed the SDS-PAGE fractionation of 24 protein, whose molecular weights (PM) ranged from 18,000 to 165,000 daltons (Da). The dogs with 1st and 2nd AHIM showed 24 protein fractions in their tracks electrophoretic, while other groups of dogs showed 23 fractions of protein, whose molecular protein weight of 68,000Da was not found. Thus, twenty-three proteins were common to proteinograms of the five experimental groups. From these, it was possible to identify eleven protein fractions nominally, and others were identified by their molecular weights. For control dogs, the anemic (groups 2, 3, 4 and 5) showed higher concentrations of serum transferrin and between them, the animals carrying the primary IMHA. All groups of dogs showed anemic levels of serum haptoglobin and phosphorylase significantly higher than the control dogs, while the serum ceruloplasmin was lower in anemic dogs. These findings provide additional information to the elucidation of the AHIMs in dogs.

RESUMO

This assay aimed to determine the serum protein - via polyacrylamide gel electrophoresis, which contained duodecil sodium sulfate (SDS-PAGE) - in 120 dogs, with different breeds and ages, seen by the Veterinary Hospital "Governor Laudo Natel." These animals were grouped into five experimental groups: Group 1 - group control with 20 dogs, group 2 - 28 dogs with regenerative anemia; group 3 - 27 dogs with arregenarative; anemia group 4 - 10 dogs with primary immune-mediated hemolytic anemia (AHIM 1.rd); group 5 - 35 dogs with secondary immune-mediated hemolytic anemia (AHIM 2.rd). The technique allowed the SDS-PAGE fractionation of 24 protein, whose molecular weights (PM) ranged from 18,000 to 165,000 daltons (Da). The dogs with 1st and 2nd AHIM showed 24 protein fractions in their tracks electrophoretic, while other groups of dogs showed 23 fractions of protein, whose molecular protein weight of 68,000Da was not found. Thus, twenty-three proteins were common to proteinograms of the five experimental groups. From these, it was possible to identify eleven protein fractions nominally, and others were identified by their molecular weights. For control dogs, the anemic (groups 2, 3, 4 and 5) showed higher concentrations of serum transferrin and between them, the animals carrying the primary IMHA. All groups of dogs showed anemic levels of serum haptoglobin and phosphorylase significantly higher than the control dogs, while the serum ceruloplasmin was lower in anemic dogs. These findings provide additional information to the elucidation of the AHIMs in dogs.

No presente protocolo experimental, determinaram-se os proteinogramas séricos, por intermédio da eletroforese em gel de poliacrilamida contendo duodecil sulfato de sódio (SDS-PAGE), de 120 cães com raças e idades variadas e atendidos junto ao Hospital Veterinário "Governador Laudo Natel" da FCAV/Unesp, com o objetivo principal de comparar diferentes frações seroproteicas em estados anêmicos regenerativos, arregenerativos, imunomediados primários e secundários. Os referidos animais foram distribuídos em cinco grupos experimentais: grupo 1: 20 cães de controle; grupo 2: 28 cães com anemia regenerativa não imune; grupo 3: 27 cães com anemia arregenerativa não imune; grupo 4: 10 cães com anemia hemolítica imunomediada primária; grupo 5: 35 cães com anemia hemolítica imunomediada secundária. A técnica SDS-PAGE permitiu o fracionamento de 24 proteínas, cujos pesos moleculares (PM) variaram de 18.000 a 165.000 daltons (Da). Os cães com AHIM primária e secundária apresentaram 24 frações proteicas em seus traçados eletroforéticos, enquanto que cães de controle (1) e portadores de anemia regenerativa (2) e arregenerativa (3) de natureza não imune apresentaram 23 frações de proteínas, cuja proteína de peso molecular 68.000Da não foi encontrada. Dessa forma, 23 frações proteicas foram detectadas e revelaram-se comuns aos proteinogramas dos cães de controle e daqueles dos quatro grupos experimentais. Destas, identificaram-se nominalmente 11 frações proteicas, e as demais foram estudadas com base nos seus respectivos pesos moleculares. Em relação aos cães de controle, os anêmicos (grupos 2, 3, 4 e 5) apresentaram maiores concentrações de transferrina sérica e entre estes os animais portadores da AHIM primária. Todos os cães anêmicos apresentaram teores séricos de haptoglobina e fosforilase significativamente maiores que os controles, enquanto que a concentração sérica de ceruloplasmina foi significativamente maior nestes. Tais achados analisados em conjunto agregam informações adicionais úteis à elucidação das AHIMs em cães.