RESUMO
Cerebral malaria (CM) is the severest form of Plasmodium falciparum infection. Children under 5 years old are those most vulnerable to CM, and they consequently have the highest risk of malaria-related death. Parasite-associated factors leading to CM are not yet fully elucidated. We therefore sought to characterize the gene expression profile associated with CM, using RNA sequencing data from 15 CM and 15 uncomplicated malaria isolates from Benin. Cerebral malaria parasites displayed reduced circulation times, possibly related to higher cytoadherence capacity. Consistent with the latter, we detected increased var genes abundance in CM isolates. Differential expression analyses showed that distinct transcriptome profiles are signatures of malaria severity. Genes involved in adhesion, excluding variant surface antigens, were dysregulated, supporting the idea of increased cytoadhesion capacity of CM parasites. Finally, we found dysregulated expression of genes in the entry into host pathway that may reflect greater erythrocyte invasion capacity of CM parasites.
Assuntos
Malária Cerebral , Malária Falciparum , Benin , Criança , Pré-Escolar , Eritrócitos/parasitologia , Perfilação da Expressão Gênica , Humanos , Malária Cerebral/metabolismo , Malária Falciparum/metabolismo , Plasmodium falciparum , Proteínas de Protozoários/metabolismo , TranscriptomaRESUMO
Inequalities between poorer and wealthier people in accessing healthcare services have been widely studied, but the mechanisms generating them are still to be fully understood. Among these, there is still a lack of evidence of relationships between health prevention/health promotion policies, welfare systems and social differences. We analysed 68 201 females from the PASSI Italian surveillance system for the years 2007-2010. The prevalence of women undergoing Pap testing was used as an example of access to preventive services. An odds ratio gradient was found with regard to different welfare system clusters: the probability of undergoing a screening test is higher for more advanced welfare systems. A strong association was found between having received a letter from the local health unit and having undergone the screening test. Significant differences still exist between high- and low-income women and their access to Italian preventive public services. As we expected, social determinants play an important role in health disparities, as these are also strongly influenced by typologies of welfare systems and by health policies.
Assuntos
Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Disparidades em Assistência à Saúde/estatística & dados numéricos , Teste de Papanicolaou/estatística & dados numéricos , Seguridade Social/estatística & dados numéricos , Adolescente , Adulto , Idoso , Detecção Precoce de Câncer , Feminino , Humanos , Itália , Pessoa de Meia-Idade , Prevenção Primária , Fatores Socioeconômicos , Adulto JovemRESUMO
PfEMP1 is the major antigen involved in Plasmodium falciparum-infected erythrocyte sequestration in cerebrovascular endothelium. While some PfEMP1 domains have been associated with clinical phenotypes of malaria, formal associations between the expression of a specific domain and the adhesion properties of clinical isolates are limited. In this context, 73 cerebral malaria (CM) and 98 uncomplicated malaria (UM) Beninese children were recruited. We attempted to correlate the cytoadherence phenotype of Plasmodium falciparum isolates with the clinical presentation and the expression of specific PfEMP1 domains. Cytoadherence level on Hbec-5i and CHO-ICAM-1 cell lines and var genes expression were measured. We also investigated the prevalence of the ICAM-1-binding amino acid motif and dual receptor-binding domains, described as a potential determinant of cerebral malaria pathophysiology. We finally evaluated IgG levels against PfEMP1 recombinant domains (CIDRα1.4, DBLß3, and CIDRα1.4-DBLß3). CM isolates displayed higher cytoadherence levels on both cell lines, and we found a correlation between CIDRα1.4-DBLß1/3 domain expression and CHO-ICAM-1 cytoadherence level. Endothelial protein C receptor (EPCR)-binding domains were overexpressed in CM isolates compared to UM whereas no difference was found in ICAM-1-binding DBLß1/3 domain expression. Surprisingly, both CM and UM isolates expressed ICAM-1-binding motif and dual receptor-binding domains. There was no difference in IgG response against DBLß3 between CM and UM isolates expressing ICAM-1-binding DBLß1/3 domain. It raises questions about the role of this motif in CM pathophysiology, and further studies are needed, especially on the role of DBLß1/3 without the ICAM-1-binding motif.IMPORTANCE Cerebral malaria pathophysiology remains unknown despite extensive research. PfEMP1 proteins have been identified as the main Plasmodium antigen involved in cerebrovascular endothelium sequestration, but it is unclear which var gene domain is involved in Plasmodium cytoadhesion. EPCR binding is a major determinant of cerebral malaria whereas the ICAM-1-binding role is still questioned. Our study confirmed the EPCR-binding role in CM pathophysiology with a major overexpression of EPCR-binding domains in CM isolates. In contrast, ICAM-1-binding involvement appears less obvious with A-type ICAM-1-binding and dual receptor-binding domain expression in both CM and UM isolates. We did not find any variations in ICAM-1-binding motif sequences in CM compared to UM isolates. UM and CM patients infected with isolates expressing the ICAM-1-binding motif displayed similar IgG levels against DBLß3 recombinant protein. Our study raises interrogations about the role of these domains in CM physiopathology and questions their use in vaccine strategies against cerebral malaria.
Assuntos
Antígenos de Protozoários/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Malária Cerebral/parasitologia , Malária Falciparum/parasitologia , Plasmodium falciparum/imunologia , Proteínas de Protozoários/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Antígenos de Protozoários/genética , Benin , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Criança , Pré-Escolar , Receptor de Proteína C Endotelial/genética , Receptor de Proteína C Endotelial/metabolismo , Eritrócitos/parasitologia , Humanos , Molécula 1 de Adesão Intercelular/genética , Malária Cerebral/fisiopatologia , Malária Falciparum/fisiopatologia , Plasmodium falciparum/genética , Plasmodium falciparum/fisiologia , Ligação Proteica , Domínios Proteicos , Proteínas de Protozoários/genéticaRESUMO
The effects of yeast supplementation on intake, production, and rumen fermentation characteristics have been widely studied, but results are inconsistent between different studies. A quantitative meta-analysis was applied to 110 papers, 157 experiments, and 376 treatments dealing with yeast supplementation in ruminants. The objective was first to highlight the major quantitative effects of live yeast supplementation on intake, rumen fermentation, and milk production, and second, to identify major differences in experimental conditions between studies that can affect the response to treatment. Some of these experimental conditions are referred to as interfering factors. Yeast supplementation increased rumen pH (+0.03 on average) and rumen volatile fatty acid concentration (+2.17 mM on average), tended to decrease rumen lactic acid concentration (-0.9 mM on average), and had no influence on acetate-to-propionate ratio. Total-tract organic matter digestibility was also increased by yeast supplementation (+0.8% on average). Yeast supplementation increased dry matter intake (DMI; +0.44 g/kg of body weight; BW), milk yield (+1.2 g/kg of BW), and tended to increase milk fat content (+0.05%), but had no influence on milk protein content. Dose effects of yeast supplementation, expressed as log(10) [1+(cfu per 100 kg of BW)], globally confirmed the qualitative effects observed in the first analysis. The positive effect of yeast supplementation on rumen pH increased with the percentage of concentrate in the diet and with the DMI level. It was negatively correlated with the level of dietary neutral detergent fiber (NDF). The positive effect of yeast supplementation on rumen volatile fatty acid concentration increased with DMI and crude protein levels. The positive effect of yeast supplementation on organic matter digestibility increased with the percentage of concentrate and NDF in the diet. The negative effect of yeast supplementation on lactic acid concentration tended to decrease when the DMI level and the percentage of concentrate in the diet increased. The effects of interfering factors were globally similar when either dose effect or qualitative effect of yeast was taken into account. Although rumen fermentation efficiency per se was not measured, these results suggest an improvement in rumen fermentation by yeast supplementation. This effect could, however, be modulated by several different factors such as DMI, percentage of concentrate or NDF in the diet, or species.
Assuntos
Dieta/veterinária , Lactação/fisiologia , Leite/metabolismo , Probióticos/administração & dosagem , Rúmen/fisiologia , Ruminantes , Saccharomyces cerevisiae/fisiologia , Animais , Bovinos , Digestão/fisiologia , Ingestão de Alimentos/fisiologia , Feminino , Conteúdo Gastrointestinal/química , Ruminantes/microbiologia , Ruminantes/fisiologiaRESUMO
This study was designed to investigate the effect of subclinical doses of T-2 toxin on liver drug-metabolizing enzymes and the immune response. Pigs were offered over a 28-day period either a control diet or diets contaminated with 540, 1324 or 2102microg pure T-2toxin/kg feed. Pigs were immunized with ovalbumin and subsequent humoral and cellular immune responses measured. Monooxygenase and transferase enzyme activities and protein expression were investigated in liver tissue samples. Pigs fed 1324 or 2102microg T-2toxin/kg feed exhibited reduced anti-ovalbumin antibody production without significant alteration to specific lymphocyte proliferation. The livers of pigs exposed to T-2 toxin presented normal cytochrome P450 content, UGT 1A and P450 2B, 2C or 3A protein expression, and glutathione- and UDP glucuronosyl-transferase activities. However, P450 1A related activities (ethoxyresorufin O-deethylation and benzo-(a)-pyrene hydroxylation) were reduced for all pigs given T-2 toxin, with P450 1A protein expression decreased in pigs fed the highest dose. In addition T-2 toxin exposure reduced certain N-demethylase activities. The results of this study confirm the immunotoxic properties of T-2 toxin, in particular toward the humoral immune response. The reduction of monooxygenase activities, even though the liver presented no tissue lesion or lipid peroxidation, suggests possible deleterious interactions of T-2 toxin with these enzymes.
Assuntos
Formação de Anticorpos/efeitos dos fármacos , Sistema Enzimático do Citocromo P-450/efeitos dos fármacos , Fígado/efeitos dos fármacos , Toxina T-2/toxicidade , Animais , Proliferação de Células/efeitos dos fármacos , Citocromo P-450 CYP1A1/efeitos dos fármacos , Citocromo P-450 CYP1A1/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Relação Dose-Resposta a Droga , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Imunização , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Masculino , Ovalbumina/imunologia , Suínos , Toxina T-2/administração & dosagemRESUMO
Moringa oleifera is a plant whose fruits, roots and leaves have been advocated for traditional medicinal uses. The physicochemical analysis shows that Moringa oleifera contains more dietary polyunsaturated fatty acids (PUFA) than saturated fatty acids (SFA). The consumption of an experimental diet enriched with Moringa oleifera extracts lowered blood pressure in spontaneously hypertensive rats (SHR), but not in normotensive Wistar-Kyoto (WKY) rats as compared to rats fed an unsupplemented control diet. Anti-CD3-stimulated T cell proliferation was diminished in both strains of rats fed the Moringa oleifera. The experimental diet lowered secretion of interleukin-2 in SHR, but not in WKY rats compared with rats fed the control diet. Studies of platelets from patients with primary hypertension and from SHR support the notion that the concentration of intracellular free calcium [Ca(2+)](i) is modified in both clinical and experimental hypertension. We observed that the basal, [Ca(2+)](i) was lower in T cells of SHR than in those of WKY rats fed the control diet. Feeding the diet with Moringa oleifera extracts to WKY rats did not alter basal [Ca(2+)](i) in T cells but increased basal [Ca(2+)](i) in SHR. Our study clearly demonstrated that Moringa oleifera exerts antihypertensive effects by inhibiting the secretion of IL-2 and modulates T cell calcium signaling in hypertensive rats.
Assuntos
Anti-Hipertensivos/uso terapêutico , Sinalização do Cálcio/efeitos dos fármacos , Hipertensão/tratamento farmacológico , Moringa oleifera , Extratos Vegetais/uso terapêutico , Linfócitos T/efeitos dos fármacos , Animais , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/farmacologia , Peso Corporal/efeitos dos fármacos , Peso Corporal/fisiologia , Sinalização do Cálcio/fisiologia , Hipertensão/imunologia , Hipertensão/fisiopatologia , Masculino , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Folhas de Planta , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Linfócitos T/fisiologiaRESUMO
OBJECTIVES: Severe Plasmodium falciparum malaria (SM) involves cytoadhesion of parasitized red blood cells, mediated by P. falciparum erythrocyte membrane protein 1, which is encoded by var genes. Expression of var gene group A and B or encoding domain cassettes DC4, DC5, DC8 and DC13 has been implicated in SM in African children, but no data exist in the context of imported malaria. The aim of this study was to investigate var gene expression linked to clinical presentation and host factors in SM imported into France. METHODS: Expression level of var gene groups A, B, C, var1, var2csa, var3 and var genes encoding DC4, DC5, DC8 and DC13 was measured by quantitative RT-PCR and expressed in transcript units. Seventy SM and 48 uncomplicated malaria (UM) P. falciparum cases were analysed according to disease severity, epidemiological characteristics (migrants or travellers) and anti-P. falciparum antibodies. Cluster analysis was performed to identify gene expression profiles. RESULTS: Var1 and B/C expression were higher in UM than SM (0.66 (0-1.1) and 1.88 (1.3-2.4); p <0.04, respectively). Group C expression differed between migrants and travellers (0.21 (0-0.75) versus 0 (0-0); p 0.002). Group A differed in naive and pre-exposed patients (1.1 (0.7-1.5) versus 0.4 (0-1.1); p 0.01). Population clusters revealed increased expression from group A and B var genes, and DC4, DC8 and DC13 in SM. CONCLUSIONS: These results corroborate the implication of DC4, DC8 and DC13 in severe imported malaria cases as African children, and their expression depends of host factors.
Assuntos
Perfilação da Expressão Gênica , Malária Falciparum/patologia , Malária Falciparum/parasitologia , Plasmodium falciparum/genética , Proteínas de Protozoários/biossíntese , Adulto , Feminino , França , Humanos , Masculino , Pessoa de Meia-Idade , Plasmodium falciparum/isolamento & purificação , Proteínas de Protozoários/genética , Reação em Cadeia da Polimerase em Tempo Real , Índice de Gravidade de Doença , Adulto JovemRESUMO
Cadmium is an important toxic environmental heavy metal. Occupational and environmental pollution with cadmium results mainly from mining, metallurgy industry and manufactures of nickel-cadmium batteries, pigments and plastic stabilizers. Important sources of human intoxication are cigarette smoke as well as food, water and air contaminations. In humans, cadmium exposures have been associated with cancers of the prostate, lungs and testes. Acute exposures are responsible for damage to these organs. Chronic intoxication is associated with obstructive airway disease, emphysema, irreversible renal failure, bone disorders and immuno-suppression. At the cellular level, cadmium affects proliferation, differentiation and causes apoptosis. It has been classified as a carcinogen by the International Agency for Research on Cancer (IARC). However, it is weakly genotoxic. Indirect effects of cadmium provoke generation of reactive oxygen species (ROS) and DNA damage. Cadmium modulates also gene expression and signal transduction, reduces activities of proteins involved in antioxidant defenses. Several studies have shown that it interferes with DNA repair. The present review focuses on the effects of cadmium in mammalian cells with special emphasis on the induction of damage to DNA, membranes and proteins, the inhibition of different types of DNA repair and the induction of apoptosis. Current data and hypotheses on the mechanisms involved in cadmium genotoxicity and carcinogenesis are outlined.
Assuntos
Cádmio/farmacologia , Cádmio/toxicidade , Reparo do DNA/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Humanos , Mutagênese , Mutagênicos/toxicidade , Estresse Oxidativo/efeitos dos fármacosRESUMO
The objectives were to determine effects of graded levels of selenized yeast derived from a specific strain of Saccharomyces cerevisiae (CNCM I-3060) on animal performance and in selenium concentrations in the blood, milk, feces, and urine of dairy cows compared with sodium selenite; and to provide preliminary data on the proportion of selenium as selenomethionine in the milk and blood. Twenty Holstein cows were used in a 5 x 5 Latin square design study in which all cows received the same total mixed rations, which varied only in source or concentration of dietary selenium. There were 5 experimental treatments. Total dietary selenium of treatment 1, which received no added selenium, was 0.15 mg/kg of dry matter, whereas values for treatments 2, 3, and 4, derived from selenized yeast, were 0.27, 0.33, and 0.40 mg/kg of dry matter, respectively. Treatment 5 contained 0.25 mg of selenium obtained from sodium selenite/kg of dry matter. There were no significant treatment effects on animal performance, and blood chemistry and hematology showed few treatment effects. Regression analysis noted significant positive linear effects of increasing dietary selenium derived from selenized yeast on selenium concentrations in the milk, blood, urine, and feces. In addition, milk selenium results indicated improved bioavailability of selenium from selenized yeast, compared with sodium selenite. Preliminary analyses showed that compared with sodium selenite, the use of selenized yeast increased the concentration of selenomethionine in the milk and blood. There was no indication of adverse effects on cow health associated with the use of selenized yeast.
Assuntos
Bovinos/metabolismo , Suplementos Nutricionais , Lactação/fisiologia , Saccharomyces cerevisiae , Selênio/administração & dosagem , Ração Animal/análise , Animais , Análise Química do Sangue/veterinária , Indústria de Laticínios/métodos , Ingestão de Alimentos/efeitos dos fármacos , Fezes/química , Feminino , Lactação/efeitos dos fármacos , Leite/química , Saccharomyces cerevisiae/química , Selênio/análise , Selênio/farmacocinética , Selenometionina/análise , Selenometionina/farmacocinética , Selenito de Sódio/administração & dosagem , Selenito de Sódio/farmacocinética , Selenito de Sódio/farmacologiaRESUMO
INTRODUCTION: In the human placenta the maternal blood circulates in the intervillous space (IVS). The syncytiotrophoblast (STB) is in direct contact with maternal blood. The wall shear stress (WSS) exerted by the maternal blood flow on the STB has not been evaluated. Our objective was to determine the physiological WSS exerted on the surface of the STB during the third trimester of pregnancy. MATERIAL AND METHODS: To gain insight into the shear stress levels that the STB is expected to experience in vivo, we have formulated three different computational models of varying levels of complexity that reflect different physical representations of the IVS. Computations of the flow fields in all models were performed using the CFD module of the finite element code COMSOL Multiphysics 4.4. The mean velocity of maternal blood in the IVS during the third trimester was measured in vivo with dynamic MRI (0.94±0.14 mm.s-1). To investigate if the in silico results are consistent with physiological observations, we studied the cytoadhesion of human parasitized (Plasmodium falciparum) erythrocytes to primary human STB cultures, in flow conditions with different WSS values. RESULTS: The WSS applied to the STB is highly heterogeneous in the IVS. The estimated average values are relatively low (0.5±0.2 to 2.3±1.1 dyn.cm-2). The increase of WSS from 0.15 to 5 dyn.cm-2 was associated with a significant decrease of infected erythrocyte cytoadhesion. No cytoadhesion of infected erythrocytes was observed above 5 dyn.cm-2 applied for one hour. CONCLUSION: Our study provides for the first time a WSS estimation in the maternal placental circulation. In spite of high maternal blood flow rates, the average WSS applied at the surface of the chorionic villi is low (<5 dyn.cm-2). These results provide the basis for future physiologically-relevant in vitro studies of the biological effects of WSS on the STB.
Assuntos
Simulação por Computador , Modelos Biológicos , Placenta/fisiologia , Estresse Mecânico , Velocidade do Fluxo Sanguíneo/fisiologia , Eritrócitos/fisiologia , Feminino , Hemodinâmica/fisiologia , Humanos , Hidrodinâmica , Placenta/irrigação sanguínea , Gravidez , Resistência ao CisalhamentoRESUMO
Human placenta extracts are widely used in clinical and fundamental research, particularly to study the hormonal and exchange functions of the placenta. However, very little is known about the distribution of the main hormone mRNAs in the placenta as a whole. Total placenta extracts are heterogeneous in their cellular components, as they contain material of both fetal and maternal origin, and in their structure. Results vary greatly depending upon the location of the biopsy and the number of biopsies performed. We used real-time quantitative RT-PCR to determine whether transcripts corresponding to the main hormones secreted by the human placenta (e.g. hCG, HPL and PGH) are equally distributed within and between term placentae. We also measured cytokeratin 7 transcripts, which are specifically expressed in the trophoblast, and transcripts corresponding to nuclear receptors PPARgamma and RXRalpha. A comparison of the results obtained with 12 different samples from each of four normal term placentae revealed that the amounts of transcripts differ considerably within and between each placenta. This emphasizes the need to study large numbers of samples when looking for significant differences in gene expression.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Placenta/metabolismo , Trofoblastos/metabolismo , Análise de Variância , Feminino , Subunidade alfa de Hormônios Glicoproteicos/genética , Hormônio do Crescimento/genética , Humanos , Queratina-7 , Queratinas/genética , PPAR gama/genética , Hormônios Placentários/genética , Lactogênio Placentário/genética , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificação , RNA Mensageiro/metabolismo , Receptor X Retinoide alfa/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Pregnancy-associated placental protein-A (PAPP-A), first isolated from maternal serum, has been identified as a metalloprotease cleaving insulin-like growth factor binding protein-4 (IGFBP-4). The source of PAPP-A during pregnancy is unclear. We therefore investigated PAPP-A expression during in vitro human villous cytotrophoblast cell (CT) differentiation into syncytiotrophoblast (ST). CT were isolated from normal first trimester, second trimester and term placentae (n=10) and cultured to form ST. PAPP-A mRNA was quantified by real-time PCR, and PAPP-A protein expression was studied by immunocytochemistry and TRACE technology with specific monoclonal antibodies. PAPP-A mRNA expression in total placental extracts increased during the course of pregnancy. PAPP-A protein was detected in the cytoplasm of both CT and ST. ST formation in vitro was associated with a 19-fold increase in PAPP-A mRNA expression and an 8-fold increase in PAPP-A secretion into the culture medium. No significant difference in PAPP-A production was observed between cultured cells isolated from early and term placentae. In conclusion, PAPP-A production in vitro, is associated to the differentiation of villous cytotrophoblast cells into syncytiotrophoblast, independently of the age of gestation.
Assuntos
Vilosidades Coriônicas/metabolismo , Proteína Plasmática A Associada à Gravidez/metabolismo , Trofoblastos/metabolismo , Adulto , Diferenciação Celular , Células Cultivadas , Primers do DNA/química , Feminino , Idade Gestacional , Humanos , Imuno-Histoquímica , Trabalho de Parto , Gravidez , Proteína Plasmática A Associada à Gravidez/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trofoblastos/citologiaRESUMO
OBJECTIVE: To limit the high number of multiple pregnancies in an IVF program. SETTING: In Vitro Fertilization Laboratory, Fertility Department, Public Hospital. INTERVENTIONS: The number of embryos transferred was limited to two instead of three. RESULTS: Limiting the number of embryos transferred to only two did not influence the take home baby rate but eliminated triplet and quadruplet gestations. Moreover, the number of patients with good quality supernumerary embryos available for cryopreservation increased. CONCLUSIONS: To reduce the high frequency of multiple gestations in an IVF program, the number of embryos replaced should be limited to a maximum of two.
Assuntos
Fertilização in vitro , Gravidez Múltipla , Criopreservação , Parto Obstétrico , Implantação do Embrião , Transferência Embrionária/métodos , Embrião de Mamíferos , Feminino , Humanos , GravidezRESUMO
The fertilizing capacity of human testicular spermatozoa and the positive outcome of an in vitro fertilization program open a wide range of opportunities for men suffering from obstructive and inoperable azoospermia. In six cases, subzonal sperm injection or intracytoplasmic sperm injection techniques were applied to inject testicular spermatozoa into human oocytes. The fertilization rate after testicular sperm injection reached 45%. Normal cleavage was observed and replacement of 10 embryos in 6 patients resulted in one chemical and one ongoing pregnancy.
Assuntos
Fertilização in vitro , Espermatozoides , Testículo/citologia , Separação Celular , Transferência Embrionária , Feminino , Humanos , Masculino , Oligospermia/fisiopatologia , GravidezRESUMO
Cell walls of yeasts and bacteria are able to complex with mycotoxins and limit their bioavailability in the digestive tract when these yeasts and bacteria are given as feed additives to animals. To identify the component(s) of the yeast cell wall and the chemical interaction(s) involved in complex formation with zearalenone, four strains of Saccharomyces cerevisiae differing in their cell wall glucan and mannan content were tested. Laboratory strains wt292, fks1, and mnn9 were compared with industrial S. cerevisiae strain sc1026. The complex-forming capacity of the yeast cell walls was determined in vitro by modelling the plots of amount of toxin bound versus amount of toxin added using Hill's model. A cooperative relationship between toxin and adsorbent was shown, and a correlation between the amount of beta-D-glucans in cell walls and complex-forming efficacy was revealed (R2 = 0.889). Cell walls of strains wt292 and mnn9, which have higher levels of beta-D-glucans, were able to complex larger amounts of zearalenone, with higher association constants and higher affinity rates than those of the fks1 and sc1026 strains. The high chitin content in strains mnn9 and fks1 increased the alkali insolubility of beta-D-glucans from isolated cell walls and decreased the flexibility of these cell walls, which restricted access of zearalenone to the chemical sites of the beta-D-glucans involved in complex formation. The strains with high chitin content thus had a lower complex-forming capacity than expected based on their beta-D-glucans content. Cooperativity and the three-dimensional structure of beta-D-glucans indicate that weak noncovalent bonds are involved in the complex-forming mechanisms associated with zearalenone. The chemical interactions between beta-D-glucans and zearalenone are therefore more of an adsorption type than a binding type.
Assuntos
Parede Celular/química , Glucanos/química , Saccharomyces cerevisiae/química , Zearalenona/química , Adsorção , Ração Animal , Animais , Disponibilidade Biológica , Qualidade de Produtos para o Consumidor , Microbiologia de Alimentos , Humanos , SolubilidadeRESUMO
Previous studies have shown that isolated beta-(1,3 and 1,6)-D-glucans and related alkali-extracted fractions from the cell wall of Saccharomyces cerevisiae are able to complex with zearalenone in vitro (affinity up to 50%) and thus may reduce the bioavailability of toxins in the digestive tract. The complexation mechanisms involve cooperative interaction between the two chemical entities that can be computed by Hill's model. Various linear or branched soluble or insoluble beta-D-glucans were evaluated to elucidate their roles in the adsorption mechanisms under three pH conditions (3.0, 6.0, and 8.0) found in the digestive tract. A constant quantity of each beta-D-glucans (1 mg/ml) was mixed at 39 degrees C with increasing amounts of zearalenone (2 to 100 microg/ml), and the amount of bound toxin was measured. Acidic and neutral conditions gave the highest affinity rates (64 to 77%) by beta-(1,3)-D-glucans, whereas alkaline conditions decreased adsorption except when beta-(1,6)-D-glucan side chains were branched on beta-(1,3)-D-glucans. Alkaline conditions appear to impede the active three dimensional conformation of beta-D-glucans and favor single helix and/or random coil structures. Study of the equilibrium between beta-D-glucan-bound and free toxins revealed that two types of chemical interactions occur during toxin complexation with beta-D-glucans, identified as weak chemical linkages such as hydrogen and van der Waals bonds.
Assuntos
Parede Celular/química , Concentração de Íons de Hidrogênio , Saccharomyces cerevisiae , Zearalenona/química , beta-Glucanas/química , Adsorção , Disponibilidade Biológica , Qualidade de Produtos para o Consumidor , Relação Dose-Resposta a Droga , Microbiologia de Alimentos , Humanos , SolubilidadeRESUMO
Fourteen out of 17 laboratories completed an interlaboratory study comparing 2 pretreatment protocols of feed samples containing authorized probiotic bacilli spores. Both methods used tryptone soy agar for enumeration. Pretreatment A involved preparation of a suspension of the feed sample in 50% ethanol. For pretreatment B, the sample was suspended in peptone salt solution and heated at 80 degrees C for 10 min. Each laboratory analyzed 12 samples (6 per pretreatment), which represented duplicates of a high (10(9) colony-forming units [CFU]/g) and low (10(5) CFU/g) level of bacilli spores or a blank that contained vegetative probiotic bacteria only. For pretreatment A, the repeatability relative standard deviation (RSD(r)) was 2.9% for the low level and 2.5% for the high. The reproducibility relative standard deviation (RSDR) values were 7.8 and 5.9%, respectively. Pretreatment B revealed RSD(r) values of 1.1 and 1.0%, and RSDR values of 5.8 and 3.4%, respectively. The heat treatment (pretreatment B) of feed samples had better precision data, resulted in higher viable bacilli counts, and was more effective in deactivating vegetative background flora. It is therefore recommended for adoption for official control purposes and for CEN and ISO standards.
Assuntos
Ração Animal/microbiologia , Bacillus/isolamento & purificação , Probióticos , Esporos Bacterianos/isolamento & purificação , Contagem de Colônia MicrobianaRESUMO
The objective of this study was to determine the concentration of total selenium (Se) and the proportions of total Se comprised as selenomethionine (SeMet) and selenocysteine (SeCys) in the postmortem tissues of female pheasants (Phasianus Colchicus Torquator) offered diets that contained graded additions of selenised-enriched yeast (SY) or a single comparative dose of sodium selenite (SS). Thiobarbituric acid reactive substances (TBARS) and tissue glutathione peroxidase (GSH-Px) activity of breast (Pectoralis Major) were assessed at 0 and 5 days postmortem. A total of 216 female pheasant chicks were enrolled into the study. Twenty-four birds were euthanased at the start of the study, and samples of blood, breast muscle, leg muscle (M. Peroneus Longus and M. Gastrocnemius), heart, liver, kidney and gizzard were collected for determination of total Se. Remaining birds were blocked by live weight and randomly allocated to one of four dietary treatments (n = 48 birds/treatment) that either differed in Se source (SY v. SS) or dose (control (0.17 mg total Se/kg), SY-L and SS-L (0.3 mg/kg total Se as SY and SS, respectively) and SY-H (0.45 mg total Se/kg)). Following 42 and 91 days of treatment, 24 birds per treatment were euthanased, and samples of blood, breast muscle, leg muscle, heart, liver, kidney and gizzard were retained for determination of total Se and the proportion of total Se comprised as SeMet or SeCys. Whole blood GSH-Px activity was determined at each time point. Tissue GSH-Px activity and TBARS were determined in breast tissue at the end of the study. There were increases in both blood and tissues to the graded addition of SY to the diet (P < 0.001), but the same responses were not apparent with the blood and tissues of selenite-supplemented birds receiving a comparable dose (SY-L v. SS-L). Although there were differences between tissue types in the distribution of SeMet and SeCys, there were few differences between treatments. There were effects of treatment on erythrocyte GSH-Px activity (P = 0.012) with values being higher in treatments SY-H and SS-L when compared with the negative control and treatment SY-L. There were no effects of treatment on tissue GSH-Px activity, which is reflected in the overall lack of any treatment effects on TBARS.
Assuntos
Suplementos Nutricionais/análise , Galliformes/metabolismo , Glutationa Peroxidase/metabolismo , Selênio/metabolismo , Criação de Animais Domésticos , Animais , Relação Dose-Resposta a Droga , Feminino , Galliformes/crescimento & desenvolvimento , Glutationa Peroxidase/sangue , Músculos Peitorais/metabolismo , Distribuição Aleatória , Selênio/sangue , Selenocisteína/sangue , Selenocisteína/metabolismo , Selenometionina/sangue , Selenometionina/metabolismo , Selenito de Sódio/sangue , Selenito de Sódio/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fermento Seco/metabolismoRESUMO
The objective of this study was to determine the concentration of total selenium (Se) and proportions of total Se comprised as selenomethionine (SeMet) and selenocysteine (SeCys) in the tissues of female turkeys offered diets containing graded additions of selenized-enriched yeast (SY), or sodium selenite (SS). Oxidative stability and tissue glutathione peroxidase (GSH-Px) activity of breast and thigh muscle were assessed at 0 and 10 days post mortem. A total of 216 female turkey poults were enrolled in the study. A total of 24 birds were euthanized at the start of the study and samples of blood, breast, thigh, heart, liver, kidney and gizzard were collected for determination of total Se. Remaining birds were blocked by live weight and randomly allocated to one of four dietary treatments (n = 48 birds/treatment) that differed either in Se source (SY v. SS) or dose (Con [0.2 mg/kg total Se], SY-L and SS-L [0.3 mg/kg total Se as SY and SS, respectively] and SY-H [0.45 mg total Se/kg]). Following 42 and 84 days of treatment 24 birds per treatment were euthanized and samples of blood, breast, thigh, heart, liver, kidney and gizzard were retained for determination of total Se and the proportion of total Se comprised as SeMet or SeCys. Whole blood GSH-Px activity was determined at each time point. Tissue GSH-Px activity and thiobarbituric acid reactive substances were determined in breast and thigh tissue at the end of the study. There were responses (P < 0.001) in all tissues to the graded addition of dietary Se, although rates of accumulation were highest in birds offered SY. There were notable differences between tissue types and treatments in the distribution of SeMet and SeCys, and the activity of tissue and erythrocyte GSH-Px (P < 0.05). SeCys was the predominant form of Se in visceral tissue and SeMet the predominant form in breast tissue. SeCys contents were greater in thigh when compared with breast tissue. Muscle tissue GSH-Px activities mirrored SeCys contents. Despite treatment differences in tissue GSH-Px activity, there were no effects of treatment on any meat quality parameter.
RESUMO
This study was conducted to determine the effects of dietary Se source and dose on metabolic and hematological profiles, and their relationships with oxidative status in horses. Twenty-five mature horses were blocked by BW and randomly allocated to 1 of 5 dietary treatments: negative control (CTRL: 0.085 mg of Se/kg of DM), 3 different dietary concentrations of organic Se provided by Se yeast (SY02, SY03, and SY04 containing 0.2, 0.3, and 0.4 mg of total Se/kg of DM, respectively), and 1 positive control provided by sodium selenite (SS03 containing 0.3 mg of total Se/kg of DM). Horses were fed the same basal diet (6 kg of grass hay and 3 kg of concentrate per horse daily) and received their respective treatments for a continuous period of 112 d. Jugular venous blood samples were collected before the morning feed on d 0, 28, 56, 84, and 112. Whole blood was analyzed for hematological profile, and plasma was analyzed for metabolites of energy, protein, and mineral metabolism; enzymatic activities and metabolites related to liver and muscle damage; and markers of inflammatory and oxidative status. Plasma metabolites related to energy, protein, and mineral metabolism, acute phase proteins, and enzyme activities related to hepatocellular, hepatobiliary, and muscle damage were not affected by Se source or dose. There were no differences among treatments in either reactive oxygen metabolites or thiol group concentrations in plasma. However, a linear decrease (P < 0. 01) in plasma total antioxidants was observed with increasing Se yeast supplementation. Furthermore, total antioxidant concentrations were less in SY03 than SS03 horses (P < 0.05), and were less in SY03 and SY04 than CTRL horses (P < 0.05). These results could be interpreted as an improvement in the preventive antioxidant systems of horses fed Se yeast. Total white blood cell count was not affected by treatment. There was a tendency for horses receiving greater concentrations of Se yeast to have greater lymphocyte counts (P = 0.09), with greater lymphocyte counts in blood of SY03 vs. SS03 horses (P < 0.05). Despite the lack of effect of Se source and dose on markers of inflammatory and liver status, the hematological profile seems to indicate an immunomodulatory action, as shown by mild changes in the white blood cell populations in response to Se yeast inclusion.