Identification of ellagic acid derivatives in methanolic extracts from Qualea species

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

Utilization of neuromuscular electrical stimulation in the acute phase of ankle immobilization at 90°: Study in rats

Objective: To evaluate the skeletal muscle glycogen content and plasmatic concentration of interleukin -6 (IL-6), interleukin-4 (IL-4), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α) in rats submitted to electrical stimulation sessions during the first three days of ankle immobilization at the position of 90°. Methods: Albinomale Wistar rats(3-4 months) were maintained in vivarium. conditions with food and water ad libitum, Submitted to 12 h photoperiodic cycles of light/dark, and distributed into 7 experimental groups (n = 6): control(C), immobilized 1 day(I1) immobilized 1 day and electrically stimulated(IE1) immobilized 2 days(12), immobilized 2 days and electrically stimulated(IE2), immobilized 3 days(13) and immobilized 3 days and electrically stimulated(IE3). Groups I utilized an acrylic resin orthesis model and groups electrically stimulated (IE) utilized the orthesis and a session of electrotherapy by a Dualpex 961 (biphasic quadratic pulse, 10 Hz, 0.4 ms, 5.0 mA, one 20 min session a day). After the experimental period, the rats were anesthetized with pentobarbital sodium(40 mg/kg) and a blood sample was colleted to evaluate the plasmatic concentration of interleukins by means of the radioimmunoassay method. The soleus and the white portion of the gastrocnemius muscle were colleted for glycogen reserves analysis(GLY). Other groups of rats were used to apply the glucose tolerance test(GTT) and insulin tolerance test(ITT). For statistical analysis, the Kolmogorov-Smirnov normality test followed by ANOVA and the Tukey tests were utilized, with a critical level established at 5%. Results: In ITT test, groups IE enhanced the skeletal muscle glucose uptake, but no changes were observed in GTT after the therapy session, which indicates that electrical stimulation is a sensibilizing method to augment skeletal muscle glucose uptake. The GLY reserves were reduced in I groups, which indicate that disuse altered insulin sensitivity and compromised energetic homeostasis. However. the IE groups displayed an augment in GLY content, suggesting that electrical stimulation restores the enzymatic pathways altered by immobilization. The improvement in GLY was accompanied by an elevation of the plasmatic concentration of IL-6 and TNF-α, showing the participation of these interleukins in the control of metabolic profile. Plasmatic concentrations of IL-10 were elevated only after 3 days of IE while IL-4 did not display any modifications. Conclusion: The results suggest that neuromuscular electricaf stimulation is an important toot in the maintenance of energetic, conditions of musculature submitted to immobilization, and presents multifactor mechanisms linked to interleukins action that converge to maintain the energetic equilibrium of the tissue in disuse.

Female sex hormones mediate the allergic lung reaction by regulating the release of inflammatory mediators and the expression of lung E-selectin in rats

Background: Fluctuations of estradiol and progesterone levels caused by the menstrual cycle worsen asthma symptoms. Conflicting data are reported in literature regarding pro and anti-inflammatory properties of estradiol and progesterone.Methods: Female Wistar rats were ovalbumin (OVA) sensitized 1 day after resection of the ovaries (OVx). Control group consisted of sensitized-rats with intact ovaries (Sham-OVx). Allergic challenge was performed by aerosol (OVA 1%, 15 min) two weeks later. Twenty four hours after challenge, BAL, bone marrow and total blood cells were counted. Lung tissues were used as explants, for expontaneous cytokine secretion in vitro or for immunostaining of E-selectin.Results: We observed an exacerbated cell recruitment into the lungs of OVx rats, reduced blood leukocytes counting and increased the number of bone marrow cells. Estradiol-treated OVx allergic rats reduced, and those treated with progesterone increased, respectively, the number of cells in the BAL and bone marrow. Lungs of OVx allergic rats significantly increased the E-selectin expression, an effect prevented by estradiol but not by progesterone treatment. Systemically, estradiol treatment increased the number of peripheral blood leukocytes in OVx allergic rats when compared to non treated-OVx allergic rats. Cultured-BAL cells of OVx allergic rats released elevated amounts of LTB4 and nitrites while bone marrow cells increased the release of TNF-α and nitrites. Estradiol treatment of OVx allergic rats was associated with a decreased release of TNF-α, IL-10, LTB4 and nitrites by bone marrow cells incubates. In contrast, estradiol caused an increase in IL-10 and NO release by cultured-BAL cells. Progesterone significantly increased TNF- α by cultured BAL cells and bone marrow cells.Conclusions: Data presented here suggest that upon hormonal oscillations the immune sensitization might trigger an allergic lung inflammation whose phenotype is under control of estradiol. Our data could contribute to the understanding of the protective role of estradiol in some cases of asthma symptoms in fertile ans post-menopausal women clinically observed. © 2010 de Oliveira et al; licensee BioMed Central Ltd.

Cardiac remodeling induced by smoking: Concepts, relevance, and potential mechanisms

Cardiac or ventricular remodeling is characterized by molecular, cellular, and interstitial alterations that lead to changes in heart size, mass, geometry and function in response to a given insult. Currently, tobacco smoke exposure is recognized as one of these insults. Indeed, tobacco smoke exposure induces the enlargement of the left-sided cardiac chambers, myocardial hypertrophy, and ventricular dysfunction. Potential mechanisms for these alterations include hemodynamic and neurohormonal changes, oxidative stress, inflammation, nitric oxide bioavailability, matrix metalloproteinases and mitogen-activated protein kinase activation. This review will focus on the concepts, relevance, and potential mechanisms of cardiac remodeling induced by tobacco smoke. © 2012 Bentham Science Publishers.

Maternal periodontal disease in rats decreases insulin sensitivity and insulin signaling in adult offspring

Background: Periodontal disease during pregnancy has been recognized as one of the causes of preterm and lowbirth- weight (PLBW) babies. Several studies have demonstrated that PLBW babies are prone to developing insulin resistance as adults. Although there is controversy over the association between periodontal disease and PLBW, the phenomenon known as programming can translate any stimulus or aggression experienced during intrauterine growth into physiologic and metabolic alterations in adulthood. The purpose of the present study is to investigate whether the offspring of rats with periodontal disease develop insulin resistance in adulthood. Methods: Ten female Wistar rats were divided into periodontal disease (PED) and control (CN) groups. All rats were mated at 7 days after induction of periodontal disease. Male offspring were divided into two groups: 1) periodontal disease offspring (PEDO; n = 24); and 2) control offspring (CNO; n = 24). Offspring body weight was measured from birth until 75 days. When the offspring reached 75 days old, the following parameters were measured: 1) plasma concentrations of glucose, insulin, fructosamine, lipase, amylase, and tumor necrosis factor-α (TNF-α); 2) insulin sensitivity (IS); and 3) insulin signal transduction (IST) in insulin-sensitive tissues. Results: Low birth weight was not detected in the PEDO group. However, plasma concentrations of glucose, insulin, fructosamine, lipase, amylase, and TNF-α were increased and IS and IST were reduced (P <0.05) in the PEDO group compared with the CNO group. Conclusion: Maternal periodontal disease may induce insulin resistance and reduce IST in adult offspring, but such alterations are not attributable to low birth weight.

Adjunctive antimicrobial photodynamic treatment of experimentally induced periodontitis in rats with ovariectomy

Background: The aim of this study is to compare antimicrobial photodynamic therapy (aPDT) as an adjunctive therapy to scaling and root planing (SRP) for the treatment of experimentally induced periodontitis in rats with ovariectomy (OVX) that are or are not treated with estrogen replacement. Methods: A total of 270 female rats were divided into three groups: 1) normal rats; 2) rats with OVX; and 3) rats with OVX with estrogen replacement. Periodontal disease was induced through the introduction of a cotton thread around the mandibular left first molar. After 7 days, the ligature was removed, and the rats were randomly divided into the following treatment groups: 1) SRP plus saline solution; 2) SRP plus low-level laser therapy (LLLT); and 3) SRP plus toluidine blue O irrigation followed by LLLT. Ten rats from each group were euthanized at days 7, 15, and 30 after dental treatment. Bone loss (BL) in the furcation region was evaluated using histometric and immunohistochemical analyses. Results: aPDT treatment resulted in reduced BL compared with SRP treatment at all time points. Additionally, rats treated with aPDT exhibited reduced numbers of tartrate-resistant acid-phosphatase-positive cells and more proliferating cell nuclear antigen-positive cells in all treatment groups regardless of estrogen status. Whereas rats treated with aPDT showed weak immunoreactivity to the receptor activator of nuclear factor-k B ligand at day 7 post-treatment, strong osteoprotegerin immunoreactivity was observed at day 15 post-treatment. Conclusion: aPDT is an effective adjunctive therapy for the treatment of periodontitis in rats with OVX that are or are not given estrogen replacement therapy.

Efeito da silibinina sobre o metabolismo oxidativo e produção de citocinas por monócitos em gestantes portadores de pré-eclâmpsia

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

Efeito da interleucina-15 sobre a atividade fungicida, metabolismo oxidativo e produção de citocinas por monócitos humanos, infectados in vitro com Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação do retinol no escarro e no soro de tabagistas e pacientes com doença pulmonar obstrutiva crônica e sua relação com o processo inflamatório

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação do efeito protetor da prolactina em linfócitos expostos a ação do metilmercúrio

O mercúrio pode ser encontrado em diversas formas, sendo a orgânica como metilmercúrio (MeHg), considerada a mais tóxica. Facilmente absorvido por via oral, se acumula na cadeia trófica e se amplifica em carnívoros aquáticos, principalmente em peixes, daí o risco maior para as populações que deles se alimentam preferencialmente, como os ribeirinhos Amazônidas. O efeito neurotóxico dessa forma de mercúrio tem sido amplamente demonstrado através de estudos epidemiológicos e experimentais. Alguns desses estudos também mostraram que hormônios e substâncias antioxidantes podem agir protegendo o organismo contra a ação deletéria do mercúrio. A prolactina é um destes hormônios que apresenta ação protetora, mas age também como citocina pró-inflamatória. Desde que o MeHg pode também agir como uma substância imunotóxica, procuramos neste trabalho estudar a ação citoprotetora da PRL em cultivos contínuos de linhagem B95-A de linfócitos de primata afim de avaliar sua fragilidade ao MeHg e sua reatividade a ação da PRL. Com o objetivo de avaliar a integridade funcional dos linfócitos expostos ao MeHg utilizou-se teste de reação colorimétrica para 3-(4,5-dimetiltiazol-2-yl)-2,5-difenil tetrazólio bromide (MTT), o qual detecta atividade metabólica mitocondrial. Para avaliar a resposta imune do linfócito, medidas da concentração do fator de necrose tumoral alfa (TNF α) no sobrenadante do cultivo, foram realizadas por ELISA. É uma citocina pró-inflamatória liberada em resposta a agressão celular de diferentes causas, incluindo estresse oxidativo, um dos efeitos agudos mais evidentes do MeHg, além disso, esta citocina também poder responder a regulação prolactinérgica em linfócitos humanos. Após 18 horas de exposição do cultivo a crescentes concentrações do metal (0,1; 1, 5, 10 e 50 μM) verificou-se significativa diminuição do tipo dose-dependente da viabilidade celular a partir de 1 μM (35%) e progressivamente até 50 μM (80%), quando poucas células íntegras foram encontradas nos cultivos. Um efeito bifásico em forma de “sino” ocorreu na liberação de TNF α, onde concentrações mais baixas de MeHg inibiram (0,1 e 1 μM), a intermediária estimulou (5 μM) e as duas maiores (10 e 50 μM) voltaram a inibir. A prolactina também diminuiu a viabilidade celular, em cerca de 30%, somente na dose mais elevada (10 nM). Por outro lado, na dose de 1 nM a PRL preveniu a diminuição de 40% da viabilidade celular resultante a exposição ao MeHg a 5 μM. Esta dose de 1 nM de PRL foi a única a estimular a liberação de TNF α, mas curiosamente, reverteu a liberação desta citocina quando associada a 5 μM de MeHg, concentração que igualmente estimulou a secreção de TNF α. Os resultados confirmaram a toxidade do MeHg para linfócitos de primatas (linhagem B95-A) e sua reversão por uma possível ação protetora da PRL. Um efeito bifásico na secreção de TNF α resultou da exposição ao MeHg, sugerindo a presença de diferentes mecanismos citotóxicos resultantes a ação mercurial. Por outro lado, a PRL foi pouco efetiva em estimular a secreção daquela citocina, invertendo esta resposta quando associada ao MeHg. No entanto, estes resultados são preliminares e carecem de um estudo mais acurado para sua completa elucidação.

Avaliação do efeito do tratamento periodontal não-cirúrgico sobre parâmetros clínicos e imunológicos em pacientes portadores de Diabetes mellitus Tipo 2: estudo clínico e imunológico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação dos marcadores clínicos e inflamatórios da doença hepática gordurosa não alcoólica em mulheres na pós-menopausa com síndrome metabólica

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Associação entre expressão de receptores TLR2 e TLR4 e produção de TNF-alpha e IL-10 por monócitos de gestantes portadoras de pré-eclâmpsia

Preeclampsia (PE) is a pregnancy specific syndrome characterized by a systemic inflammatory response, with higher intensity than that observed in normal pregnancy. Cells of the immune system, such as monocytes and granulocytes are endogenously activated and secrete high levels of free radicals and inflammatory cytokines. The objective of this study was to assess the activation state of monocytes from pregnant women with preeclampsia by endogenous expression of TLR2 e TLR4 receptors and to correlate the expression of TLR2 and TLR4 on monocytes surface of pregnant women with PE with the production of tumor necrosis factor-alpha (TNF- and interleukin-10 (IL-10) by these cells stimulated or not with peptidoglycan (PG) and lipopolysaccharide (LPS), as agonists agents of TLR2 and TLR4, respectively. We evaluated 15 pregnant women with PE, 15 normotensive pregnant women (NT) and 15 non-pregnant (NP). Peripheral blood monocytes were incubates in the presence or absence of LPS or PG. The supernatant obtained after 18h of culture was aspirated and used for TNF- and IL-10 determination by enzyme immunoassay (ELISA). The endogenous expression of TLR2 and TLR4 receptors was evaluated by flow cytometry. Our results showed significant highly concentrations of TNF- and TLR4 expression in monocytes of preeclamptic women when compared with NT and NP. Normal pregnant women presented higher levels of IL-10 in comparison with PE and NP groups. TLR2 expression was similar in the three groups studied. Therefore, our study highlights the important role of TLR4 in PE and the consequent high production of TNF- by monocytes of these patients, as well as the potential mechanism involving low levels of IL-10 in the pathophysiology of the disease. These observations demonstrate the strong link between the pathology of PE and the immune system of these patients

Síntese e avaliação biológica de novos derivados anti-inflamatórios esteroides

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Resposta de fibroblastos da gengiva de pacientes jovens e idosos e efeito da laserterapia de baixa potência e de fator de crescimento sobre estas células

Pós-graduação em Reabilitação Oral - FOAR