Ativação local da rota da quinurenina por moduladores inflamatórios durante a meningite bacteriana.

Activation of the kynurenine (KYN) pathway (KP) by modulators of immune system has been observed during several neurological diseases. Here we assessed the association of chemo-/cytokine levels with the concentration of KP metabolites in cerebrospinal fluid (CSF) and plasma samples from patients with bacterial meningitis (BM). All samples were collected from 42 patients diagnosed with acute bacterial meningitis (ABM), aseptic meningitis, tuberculous meningitis and patients without infection neurological disorders. CSF and plasma concentration of metabolites from the KP was assessed by high pressure liquid chromatography (HPLC) and cytokines and chemokines by Bio-plex 200 suspension array system. Concentrations of the KP metabolites KYN and kynurenic acid (KYNA) were significantly higher in CSF of patients with ABM compared to other groups. Tryptophan (TRP), anthranilic acid (AA), 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) did not show statistical significance, although some of them presented a good accumulation during ABM. The expression of TNF-alpha, IL-6, IL-1beta, IFN-gamma, IL-10, IL-1 receptor antagonist (IL-1Ra), MIP-1alpha, MIP-1beta, MCP-1 and G-CSF was about 100-fold higher in CSF from ABM patients than other infected groups. In all CSF and plasma samples, the concentration of IL-2, IL-12(p70), IL-4, IL-8 and GM-CSF was not significant. ABM still showed significant concentrations of IL-6, IL-10, IL-1Ra and MCP-1 in plasma samples. Based on the comparison of KP metabolites concentrations between plasma and CSF samples we conclude that the activation of the tryptophan pathway upon BM occurs within the brain. This increase in KP metabolites is most due to activation of the KP by molecules as IFN-gamma and TNF-alpha in response to infection.

Análise das mudanças no perfil protéico durante o estresse oxidativo in vivo e atuação da MutY-Glicosilase em respostas celulares

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Análise imuno-histoquímica das metaloproteinases da matriz ( MMP-1,MMP-2 e MMP-9) na doença periodontal

The tissular destruction found in periodontal diseases is caused mainly by components of the host that have its production stimulated by the products of the microorganisms present on the plaque. Matrix Metalloproteinases (MMPs), a class of enzymes involved both in physiologic and pathologic extracellular matrix degradation are considered the main responsible for the characteristic tissular loss in periodontal disease, and the understanding of how this happens can have a series of beneficial implications for prevention, diagnosis and treatment of this illness. The aim of this work was to study the immunohistochemical expression of MMP-1, MMP-2, and MMP-9 in fragments of gingival biopsies with clinical diagnose of periodontal disease. MMP-1 has expressed significantly more than the others MMPs in gingivitis both in the epithelium (p=0,0008) and connective tissue (p=0,0049). In periodontitis, both MMP-1 and MMP-9 has expressed significantly more than MMP-2 in the epithelium (p<0,0001) and in the connective tissue (p=0,0002). MMP-1 and MMP-9 presented more expression in periodontitis than in gingivitis but MMP-1 only in the connective tissue (p=0,03) and MMP-9 in the epithelium (p=0,003) and in the connective tissue (p=0,04). In conclusion, these results indicate that the MMP-1 presents high expression in every stages of the periodontal diseases, and increases its expression in the connective tissue when the gingivitis evolves to periodontitis. Therefore, it may have an important role in connective tissue degradation and bone loss observed in disease, since early, in gingivitis, until late stages, in periodontitis, of the periodontal disease. MMP-9 has expressed more in periodontitis than in gingivitis, both in epithelium and in connective tissue. It means that this enzyme may have some importance in the progression of gingivitis to periodontitis by acting in bone resorption observed in this desease

Os núcleos dopaminérgicos do mesencéfalo do mocó (kerodon rupestris): caracterização citoarquitetônica e por imunoistoquímica para tirosina-hidroxilase

The 3-hydroxytyramine/dopamine (DA) is a monoamine of catecholamineric group and consists in the progenitor substantia of synthesis of noradrenaline and adrenaline, having the enzyme tyrosine hydroxylase as a regulator of this process. Nuclei of midbrain expressing DA are the retrorubral field (RRF, A8 group), the substantia nigra pars compacta (SNc, A9 group) and the ventral tegmental area (VTA, A10 group). These nuclei are involved in three complex circuitry called mesostriatal, mesocortical and mesolimbic, which are related directly with various behavioral manifestations such as motor control, reward signaling in behavioural learning, motivation and pathological manifestations of Parkinson s disease and schizophrenia. The aim of this study was describe the morphology of midbrain dopaminergic neurons (A8, A9 and A10) of the rock cavy (Kerodon rupestris), a rodent belonging to the family Caviidae typical of the Brazilian Northeast, which is being adopted as a model for neuroanatomical studies in laboratory of neuroanatomy of the Federal University of Rio Grande do Norte. Coronal sections of brains of the rock cavies were submitted to staining by Nissl s method and immunohistochemistry against tyrosine hydroxylase. The nuclear organization of the midbrain dopaminergic nuclei of the rock cavy is very similar to that found in other animals of the order Rodentia, except by the presence of the tail of substantia nigra, which was found only in the studied species. We concluded that the midbrain dopaminergic nuclei are phylogenetically stable among species, but we think to be it necessary to expand the studies about the particularity found the rock cavy, investigating its occurrence in other species of rodents or investigating its functional relevance

Participação do circuito dopaminérgico nas alterações do comportamento de medo inato de camundongos infectados pelo Toxoplasma gondii

The protozoan parasite Toxoplasma gondii transforms the innate aversion of rats for cat urine into a fatal attraction, that increases the likelihood of the parasite completing its life cycle in the cat s intestine. The neural circuits implicated in innate fear, anxiety, and learned fear all overlap considerably, raising the possibility, that T. gondii may disrupt all of these nonspecifically. In this study, we evaluated immunoreactivity for tyrosine hydroxylase (TH) in areas associated with innate fear of infected male swiss mice. The latent Toxoplasma infection converted the aversion of mice to feline odors into attraction. This loss of fear is remarkably specific, as demonstrated by Vyas et al (2007), because infection did not diminish learned fear, anxiety-like behavior, olfaction, or nonaversive learning. However, the neurochemical mechanism related to alterations in innate fear due to T. gondii infection remains poorly studied. 20 mice were inoculated with bradyzoites (25 cysts) from a Toxoplasma gondii (Me-49 strain). The brains were removed after 60 days, sectioned and processed for TH immunohistochemistry. The correlation between the amount of cysts per area and the densitometric analysis of neurotransmitter reactivity was low in the areas implicated in innate fear of infected animals, when comparated with noninfected controls

Participação da neurotransmissão dopaminergica no efeito hiperlocomotor do neuropeptideo S

Neuropeptide S (NPS) is an endogenous 20-aminoacid peptide which binds a G protein-coupled receptor named NPSR. This peptidergic system is involved in the modulation of several biological functions, such as locomotion, anxiety, nociception, food intake and motivational behaviors. Studies have shown the participation of NPSR receptors in mediating the hyperlocomotor effects of NPS. A growing body of evidence suggests the participation of adenosinergic, dopaminergic and CRF systems on the hyperlocomotor effects of NPS. Considering that little is known about the role of dopaminergic system in mediating NPS-induced hyperlocomotion, the present study aims to investigate the locomotor actions of intracerebroventricular (icv) NPS in mice pretreated with α-metil-p-tirosine (AMPT, inhibitor of dopamine synthesis), reserpine (inhibitor of dopamine vesicle storage) or sulpiride (D2 receptor antagonist) in the open field test. A distinct group of animals received the same pretreatments described above (AMPT, reserpine or sulpiride) and the hyperlocomotor effects of methylphenidate (dopamine reuptake inhibitor) were investigated in the open field. NPS and methylphenidate increased the mouse locomotor activity. AMPT per se did not change the locomotion of the animals, but it partially reduced the hyperlocomotion of methylphenidate. The pretreatment with AMPT did not affect the psychostimulant effects of NPS. Both reserpine and sulpiride inhibited the stimulatory actions of NPS and methylphenidate. These findings show that the hyperlocomotor effects of methylphenidate, but not NPS, were affected by the pretreatment with AMPT. Furthermore, methylphenidate- and NPS-induced hyperlocomotion was impaired by reserpine and sulpiride pretreatments. Together, data suggests that NPS can increase locomotion even when the synthesis of catecholamines was impaired. Additionally, the hyperlocomotor effects of NPS and methylphenidate depend on monoamines vesicular storaged, mainly dopamine, and on the activation of D2 receptors. The psychostimulant effects of NPS via activation of dopaminergic system display clinical significance on the treatment of diseases which involves dopaminergic pathways, such as Parkinson s disease and drug addiction

Estudo da imobilização de proteases para a síntese de oligolisinas

Enzymatic synthesis of peptides using proteases has attracted a great deal of attention in recent years. One key challenge in peptide synthesis is to find supports for protease immobilization capable of working in aqueous medium at high performance, producing watersoluble oligopeptides. At present, few reports have been described using this strategy. Therefore, the aim of this thesis was to immobilize proteases applying different methods (Immobilization by covalent bound, entrapment onto polymeric gels of PVA and immobilization on glycidil metacrylate magnetic nanoparticles) in order to produce water-soluble oligopeptides derived from lysine. Three different proteases were used: trypsin, α-chymotrypsin and bromelain. According to immobilization strategies associated to the type of protease employed, trypsin-resin systems showed the best performance in terms of hydrolytic activity and oligopeptides synthesis. Hydrolytic activities of the free and immobilized enzymes were determined spectrophotometrically based on the absorbance change at 660 nm at 25 °C (Casein method). Calculations of oligolysine yield and average degree of polymerization (DPavg) were monitored by 1H-NMR analysis. Trypsin was covalently immobilized onto four different resins (Amberzyme, Eupergit C, Eupergit CM and Grace 192). Maximum yield of bound protein was 92 mg/g, 82 mg/g and 60 mg/g support for each resin respectively. The effectiveness of these systems (Trypsin-resins) was evaluated by hydrolysis of casein and synthesis of water-soluble oligolysine. Most systems were capable of catalyzing oligopeptide synthesis in aqueous medium, albeit at different efficiencies, namely: 40, 37 and 35% for Amberzyme, Eupergit C and Eupergit CM, respectively, in comparison with free enzyme. These systems produced oligomers in only 1 hour with DPavg higher than free enzyme. Among these systems, the Eupergit C-Trypsin system showed greater efficiency than others in terms of hydrolytic activity and thermal stability. However, this did not occur for oligolysine synthesis. Trypsin-Amberzyme proved to be more successful in oligopeptide synthesis, and exhibited excellent reusability, since it retained 90% of its initial hydrolytic and synthetic activity after 7 reuses. Trypsin hydrophobic interactions with Amberzyme support are responsible for protecting against strong enzyme conformational changes in the medium. In addition, the high concentration of oxirane groups on the surface promoted multi-covalent linking and, consequently, prevented the immobilized enzyme from leaching. The aforementioned results suggest that immobilized Trypsin on the supports evaluated can be efficiently used for oligopeptides synthesis in aqueous media

Insuficiência cardíaca com fração de ejeção normal

A insuficiência cardíaca com fração de ejeção normal (ICFEN) é uma síndrome complexa que vem sendo largamente estudada, desde a última década. É causada por disfunção ventricular diastólica evidenciada por métodos complementares, como estudo hemodinâmico ou ecocardiograma, na presença de fração de ejeção preservada. Acomete preferencialmente indivíduos mais idosos e com comorbidades, como hipertensão arterial sistêmica, insuficiência coronariana e obesidade. Os mecanismos fisiopatológicos são complexos e multifatoriais, envolvendo a rigidez passiva do miocárdio, a geometria ventricular, a força de contenção do pericárdio e a interação entre os ventrículos. Os objetivos principais do tratamento são reduzir a congestão venosa pulmonar, a frequência cardíaca e controlar as comorbidades. Ainda não há evidências fortes de que o uso de medicações específicas, como inibidores de enzima de conversão ou betabloqueadores, interfiram na mortalidade. Os fatores de pior prognóstico incluem a idade avançada, presença de disfunção renal, diabete, classe funcional III e IV (NYHA) e estágio avançado de disfunção diastólica, com padrão restritivo ao enchimento ventricular. Outro aspecto que vem ganhando espaço na literatura é o questionamento do papel da disfunção sistólica nos quadros de ICFEN. Todos esses aspectos são abordados detalhadamente na presente revisão.

AT1-receptor mediated vascular damage in myocardium, kidneys and liver in rats

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

Atividade antiinflamatória de uma heterofucana da alga marrom Padina gymnospora

Fucans, sulfated polysaccharides extracted from brown algae and some echinoderms, have been extensively studied for its diverse biological activities and because of its interference with molecular mechanisms of cell to cell recognition, including leukocyte trafficking from blood vessels into sites of inflammation mediated by selectin, a family of adhesion molecules. In the present study, we examined structural features of a heterofucan extracted from brown algae Padina gymnospora and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolysis with the proteolytic enzyme maxatase. The presence of protein and uronic acid contamination was detected in the crude polysaccharide extract. Fractionation of the crude extract with growing concentrations of acetone produced five fractions with different concentrations of fucose, xylose, uronic acid, galactose, glucose and sulfate. The fraction precipitated with 1.5 volumes of acetone was characterized by infrared and nuclear magnetic resonance, through which can be observed the presence of sulfate groups in the C4 of -L-fucose. The anti-inflammatory action of this composite was assessed by a sodium thioglycollate-induced peritonitis assay and through nitric oxide production by the peritoneal macrophages using Griess reagent. Fraction F1.5 was efficient in reducing leukocyte influx into the peritoneal cavity when 10 mg/kg and 25mg/kg were used, resulting in a decrease of 56 and 39%, respectively. A decrease of nitric oxide production occurred when high concentrations of fucana were used. The cytotoxicity of the composite was also assessed using the reduction of 3-(4,5 dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT). Fraction F1.5 had no cytotoxicity when 500 μg/mL of the fraction was used. This study suggests the use of fraction F1.5 (heterofucan) as an anti-inflammatory

Avaliação da concentração de pectinase no processo de hidrólise-sacarificação do farelo de mandioca para obtenção de etanol

Neste trabalho objetivou-se avaliar a concentração de pectinase (Pectinex Ultra SP-L) no processo de hidrólise-sacarificação do farelo de mandioca para produção de etanol. Foram avaliadas quatro concentrações da enzima pectinase com enzima complemetar as amilases e o tratamento com apenas as amilases. Realizou-se a caracterização do hidrolisado e resíduo fibroso resultantes do processo, e os resultados obtidos demonstraram que a concentração mínima de pectinase para um bom rendimento do processo foi 8Kg enzima/t fibras, com. 89,4% do amido hidrolisado. Quanto ao resíduo fibroso, este apresenta potencialidade de aproveitamento com base para produtos dietéticos ricos em fibras.

Uso de medicamentos por pessoas com deficiências em áreas do estado de São Paulo

OBJETIVO: Analisar o consumo de medicamentos e os principais grupos terapêuticos consumidos por pessoas com deficiências físicas, auditivas ou visuais. MÉTODOS: Estudo transversal em que foram analisados dados do Inquérito Multicêntrico de Saúde no Estado de São Paulo (ISA-SP) em 2002 e do Inquérito de Saúde no Município de São Paulo (ISA-Capital), realizado em 2003. Os entrevistados que referiram deficiências foram estudados segundo as variáveis que compõem o banco de dados: área, sexo, renda, faixa etária, raça, consumo de medicamentos e tipos de medicamentos consumidos. RESULTADOS: A percentagem de consumo entre as pessoas com deficiência foi de: 62,8% entre os visuais; 60,2% entre os auditivos e 70,1% entre os físicos. As pessoas com deficiência física consumiram 20% mais medicamentos que os não-deficientes. Entre as pessoas com deficiência visual, os medicamentos mais consumidos foram os diuréticos, agentes do sistema renina-angiotensina e analgésicos. Pessoas com deficiência auditiva utilizaram mais analgésicos e agentes do sistema renina-angiotensina. Entre indivíduos com deficiência física, analgésicos, antitrombóticos e agentes do sistema renina-angiotensina foram os medicamentos mais consumidos. CONCLUSÕES: Houve maior consumo de medicamentos entre as pessoas com deficiências quando comparados com os não-deficientes, sendo os indivíduos com deficiência física os que mais consumiram fármacos, seguidos de deficientes visuais e auditivos.

Níveis séricos de enzimas de função hepática em frangos de corte de criação industrial clinicamente saudáveis

The values for the main hepatic enzymes included in the profiles of screen clinical biochemistry, alanine-aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (FA), lactate desidrogenase (LDH) and gamaglutamiltransferase (GGT), in samples of serum of broiler chickens in industrial system, clinically healthy, starting from the seventh day of life, until the slaughter (42 days) in weekly intervals were determined. Significant variations were not observed in the analyses in relation to the age of the birds for none of the appraised enzymes.

Efeitos da suplementação de fitase sobre a disponibilidade aparente de Mg, Ca, Zn, Cu, Mn e Fe em alimentos vegetais para a tilápia-do-nilo

Cem juvenis de tilápia-do-nilo (Oreochromis niloticus; PV = 100.0 ± 5.0 g) foram distribuídos em 10 tanques-rede com o objetivo de avaliar o efeito da suplementação da enzima fitase (0, 1.000 e 2.000 UFA/kg) sobre a disponibilidade de minerais em alguns alimentos energéticos (milho, milho extrusado, farelo de trigo, farelo de arroz e farelo de sorgo) e protéicos (farelo de soja extrusado, farelo de soja, farelo de girassol, farelo de algodão e glúten de milho) utilizados na alimentação de tilápia-do-nilo (Oreochromis niloticus). Para determinação dos coeficientes de disponibilidade aparente (CDA) do cálcio (Ca), magnésio (Mg), zinco (Zn), cobre (Cu), ferro (Fe) e manganês (Mn), foram confeccionadas 31 rações, marcadas com 0,10% de óxido de crômio III uma referência (ração purificada) e 30 contendo os dez alimentos e os diferentes níveis de suplementação da enzima fitase. O CDA dos nutrientes foi calculado com base no teor de crômio da ração e das fezes. A fitase aumenta, nos vegetais, a disponibilidade do Mg, Cu, Zn e Mn, os quais apresentam tendência diferenciada, em razão do seu valor biológico e do nível de suplementação de enzima.

Digestibilidade aparente de rações contendo complexo enzimático para tilápia-do-nilo

Avaliou-se a digestibilidade aparente de cinco rações isoproteicas e isoenergéticas contendo 0; 0,1; 0,2; 0,3 e 0,4g do complexo enzimático Bioenzimaplus (lipase, protease e carboidrase)/kg de ração na alimentação da tilápia-do-nilo. Foram utilizados 100 peixes distribuídos em cinco aquários de alimentação e cinco aquários de coleta de fezes. Os cinco tratamentos foram arranjados em delineamento inteiramente ao acaso com cinco repetições. Houve efeito linear de tratamento sobre a digestibilidade aparente (CDa) para proteína bruta e extrato etéreo. O CDa da proteína bruta e do extrato etéreo variaram de 81,60% a 84,93% e de 74,19% a 82,69%, respectivamente. Para carboidrato e energia bruta, o maior nível de suplementação resultou em maior CDa, 57,85% e 63,78%, respectivamente( P<0,05). Não foi observada diferença entre tratamentos no CDa da matéria seca, apresentando valores de 49,47% a 58,55%. A inclusão de complexo enzimático - lipase, protease e carboidrase - em dietas para tilápia-do-nilo melhorou a digestibilidade da proteína, do extrato etéreo, do carboidrato e da energia das rações.