The cytoprotective effect of alpha-tocopherol and daidzein against d-galactosamine-induced oxidative damage in the rat liver

We hypothesized that the hepatotoxicity that develops after the induction of oxidative stress (induced by d-galactosamine [GalN]) can be ameliorated by alpha-tocopherol (ATC) and the soy isoflavone daidzein. To test this, we ranked and assigned male Wistar rats into 6 groups, which involved pretreatment (ATC or daidzein) for 1 hour followed by treatment (GalN) for 23 hours. Histopathologic analysis showed that GalN administration induced marked necrosis (P < .001), steatosis (P < .001), both lobular and portal inflammations (P < .001), overall histopathologic score (P < .001), and activation of caspase-3 in the liver (P < .001). Immunohistochemical staining of malondialdehyde-protein adducts, a measure of oxidative stress, was increased in response to GalN (P < .001). Paradoxically, there were increases in total (P < .05) and cytosolic superoxide dismutase (P < .001) activities after GalN administration, indicative of an up-regulation of antioxidant defenses. The concentration of total protein (P < .001), albumin (P < .01), and globulin fractions (P < .001) in the plasma, as well as the activity of aspartate aminotransferase (P < .001), was significantly perturbed after GalN treatment, reflective of overall acute hepatic injury. Administration of daidzein showed a significant amelioration of the Ga1N-induced increase in malondialdehyde-protein adducts (P < .01) and cytosolic superoxide dismutase activities (P < .01) in the liver. However, all other variables were not significantly altered in response to daidzein. In response to ATC pretreatment, the total histopathologic score (P < .05), degree of necrosis (P < .05), and both lobular (P < .05) and portal (P = .05) inflammations were significantly ameliorated. To conclude, both daidzein and ATC protect the liver against oxidative damage possibly via different pathways.

Immobilized Kidney 28-kDa Endostatin- Related (KES28kDa) Fragment Promotes Endothelial Cell Survival

Background/Objective: Renal ischemia-hypoxia is a leading cause of acute kidney injury (AKI). Ischemia causes extracellular matrix breakdown of the tubular basement membrane. Endostatin (ES) is the C-terminal fragment of collagen XVIII generated by proteolytic cleavage. Recent studies have demonstrated that ES expression is upregulated in ischemic kidneys. The present study aimed to characterize ES from ischemic kidneys. Methods: Ischemic renal failure was induced via 45 min of occlusion of the left renal artery and vein. After the ischemic period, blood was collected. Kidneys were harvested and used for immunohistochemical testing and protein extraction. Three-step purification was used. Soluble and immobilized purified ES were tested in cell viability and adhesion assays. Results: The soluble KES28kDa inhibited endothelial cell proliferation: 25 versus 12.5 mu g (p < 0.05); 12.5 versus 3.15 mu g (p < 0.05). Immobilization of KES28kDa supports endothelial cell survival over the control p = 0.021). Human umbilical vein endothelial cells plated on immobilized KES28kDa showed an increase in membrane ruffles and stress fibers. Conclusion: These data demonstrate the local synthesis of a 28-kDa ES-related fragment following AKI and suggest its role in endothelium survival. Copyright (C) 2010 S. Karger AG, Basel

The Immunomodulatory Action of Sialostatin L on Dendritic Cells Reveals Its Potential to Interfere with Autoimmunity

Sialostatin L (SialoL) is a secreted cysteine protease inhibitor identified in the salivary glands of the Lyme disease vector Ixodes scapularis. In this study, we reveal the mechanisms of SialoL immunomodulatory actions on the vertebrate host. LPS-induced maturation of dendritic cells from C57BL/6 mice was significantly reduced in the presence of SialoL. Although OVA degradation was not affected by the presence of SialoL in dendritic cell cultures, cathepsin S activity was partially inhibited, leading to an accumulation of a 10-kDa invariant chain intermediate in these cells. As a consequence, in vitro Ag-specific CD4(+) T cell proliferation was inhibited in a time-dependent manner by SialoL, and further studies engaging cathepsin S(-/-) or cathepsin L(-/-) dendritic cells confirmed that the immunomodulatory actions of SialoL are mediated by inhibition of cathepsin S. Moreover, mice treated with SialoL displayed decreased early T cell expansion and recall response upon antigenic stimulation. Finally, SialoL administration during the immunization phase of experimental autoimmune encephalomyelitis in mice significantly prevented disease symptoms, which was associated with impaired IFN-gamma and IL-17 production and specific T cell proliferation. These results illuminate the dual mechanism by which a human disease vector protein modulates vertebrate host immunity and reveals its potential in prevention of an autoimmune disease. The Journal of Immunology, 2009, 182: 7422-7429.

Deficiency of thrombospondin-1 reduces Th17 differentiation and attenuates experimental autoimmune encephalomyelitis

Transforming growth factor beta (TGF-beta) plays a role both in the induction of Treg and in the differentiation of the IL-17-secreting T cells (Th17) which drive inflammation in experimental autoimmune encephalomyelitis (EAE). We investigated the role that thrombospondin-1 (TSP-1) dependent activation of TGF-beta played in the generation of an encephalitic Th17 response in EAE. Upon immunization with myelin oligodendrocyte glycoprotein peptide (MOG(35-55)), TSP-1 deficient (TSP-1(null)) mice and MOG(35-55) TCR transgenic mice that lack of TSP-1 (2D2.TSP-1(null)) exhibited an attenuated form of EAE, and secreted lower levels of IL-17. Adoptive transfer of in vitro-activated 2D2.TSP-1(null) T cells induced a milder form of EAE, independent of TSP-1 expression in the recipient mice. Furthermore, in vitro studies demonstrated that anti-CD3/anti-CD28 pre-activated CD4+ T cells transiently upregulated latent TGF-beta in a TSP-1 dependent way, and such activation of latent TGF-beta was required for the differentiation of Th17 cells. These results demonstrate that TSP-1 participates in the differentiation of Th17 cells through its ability to activate latent TGF-beta, and enhances the inflammatory response in EAE. (C) 2009 Elsevier Ltd. All rights reserved.

Association of Human T Lymphotropic Virus 1 Amplification of Periodontitis Severity with Altered Cytokine Expression in Response to a Standard Periodontopathogen Infection

Background. Periodontal diseases (PDs) are infectious diseases in which periodontopathogens trigger chronic inflammatory and immune responses that lead to tissue destruction. Recently, viruses have been implicated in the pathogenesis of PDs. Individuals infected with human T lymphotropic virus 1 (HTLV-1) present with abnormal oral health and a marked increased prevalence of periodontal disease. Methods. In this study, we investigated the patterns of periodontopathogen infection and local inflammatory immune markers in HTLV-1-seropositive individuals with chronic periodontitis (CP/HTLV-1 group) compared with HTLV-1 -seronegative individuals with chronic periodontitis (CP group) and periodontally healthy, HTLV-1 -seronegative individuals (control group). Results. Patients in the CP/HTLV-1 group had significantly higher values of bleeding on probing, mean probing depth, and attachment loss than patients in the CP group. The expression of tumor necrosis factor a and interleukin (IL) 4 was found to be similar in the CP and CP/HTLV-1 groups, whereas IL-12 and IL-17 levels trended toward a higher expression in the CP/HTLV-1 group. A significant increase was seen in the levels of IL-1 beta and interferon gamma in the CP/HTLV-1 group compared with the CP group, whereas expression of the regulatory T cell marker FOXp3 and IL-10 was significantly decreased in the lesions from the CP/HTLV-1 group. Interestingly, similar frequency and/or load of periodontopathogens (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans) and frequency of viruses (herpes simplex virus 1, human cytomegalovirus, and Epstein-Barr virus) characteristically associated with PDs were found in the CP/HTLV and CP groups. Conclusions. HTLV-1 may play a critical role in the pathogenesis of periodontal disease through the deregulation of the local cytokine network, resulting in an exacerbated response against a standard periodontopathogen infection.

Efeitos do alcalóide indólico caulerpina, obtido de alga do gênero caulerpa, em modelo de colite experimental murina

As doenças inflamatórias intestinais são enfermidades onde a tolerância e homeostase da resposta inflamatória estão comprometidas, gerando lesões teciduais e favorecendo o surgimento de neoplasias. Há dois importantes exemplos de doenças inflamatórias intestinais, a colite ulcerativa, foco do modelo desse estudo, e a doença de Crohn. Os medicamentos utilizados no tratamento dessas doenças desencadeiam diversos efeitos adversos; além disso, em alguns pacientes, eles não são eficazes. Os extratos de algas têm demonstrado várias atividades biológicas, entre elas a atividade anti-inflamatória. Os extratos das algas do gênero Caulerpa foram utilizados em vários estudos, onde modelos inflamatórios foram analisados, entre eles o modelo de colite ulcerativa. A utilização do extrato metanólico da C. mexicana como terapêutica nesse modelo atenuou o quadro clinico desenvolvido pelos animais. Sendo assim, o presente estudo teve como objetivo analisar a ação terapêutica da Caulerpina (CLP), extraída da C. racemosa, no modelo murino de colite ulcerativa. Camundongos C57BL/6 machos foram expostos a uma solução de Dextrana Sulfato de Sódio (DSS) a 3% por sete dias. A partir do primeiro dia de exposição ao DSS os animais foram tratados em dias alternados com a CLP nas doses de 4 e 40 mg/kg e com a dexametasona (3 mg/kg) por via oral. O desenvolvimento da doença foi analisado através do índice de atividade da doença (IAD), que leva em consideração a perda de peso corporal, a consistência e a presença de sangue nas fezes. Após a eutanásia, o cólon foi removido e mensurado, e amostras do tecido colônico foram destinadas a análise histológica e à cultura para dosagem de citocinas. Os níveis de citocinas no sobrenadante da cultura do cólon foram mensurados por ELISA. O tratamento com a CLP (4 mg/kg) desencadeou significativa melhora quanto à perda de peso corporal e ao IAD, e atenuou o encurtamento do cólon em resposta ao DSS. Tal dose foi capaz de reduzir os níveis de citocinas pró-inflamatórias analisadas (TNF-, IFN-, IL-6, IL-17), mas não teve efeito significativo nas citocinas anti-inflamatórias IL-10 e TGF-. O tratamento com a CLP (40 mg/kg) não foi eficaz quanto a perda de peso e ao IAD, além de não ter atenuado a redução do cólon em resposta ao DSS. Essa dose conseguiu reduzir os níveis das citocinas pró-inflamatórias, porém não os níveis de IL-6. O tratamento com a dexametasona obteve melhora discreta quanto à perda de peso corporal e ao IAD, porém não atenuou a redução do cólon em resposta ao DSS. Esse tratamento também conseguiu reduzir todas as citocinas pró-inflamatórias testadas. Deste modo a CLP (4 mg/kg) demonstrou ser uma alternativa promissora no tratamento da colite ulcerativa, em razão dos seus efeitos benéficos sobre parâmetros clínicos, morfológicos e moleculares do modelo em estudo

Dendritic cell are able to differentially recognize Sporothrix schenckii antigens and promote Th1/Th17 response in vitro

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

Leukotrienes are not essential for the efficacy of a heterologous vaccine against Mycobacterium tuberculosis infection

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

Análise da resposta th17 em líquen plano oral

Th17 cells have been strongly associated with the pathogenesis of several autoimmune and inflammatory diseases. IL-17 and IL-23 are important cytokines associated with this lineage. The aim of this study was to analyze, through immunohistochemical methods, the immunoexpression of IL-17 and IL-23 in the inflammatory infiltrate of oral lichen planus (OLP) lesion compared to that of inflammatory fibrous hyperplasia (IFH) and between clinical forms reticular and erosive of OLP. The sample included 41 cases of OLP, of which 23 were reticular and 18 erosive and 10 cases of IFH. The results were subjected to nonparametric statistical tests with a 5% significance level. In OLP lesions histomorphological analysis, the most common findings were: hyperparakeratinization, specimens with atrophic epithelium in erosive clinical form (p = 0.011), epithelial projections in most of reticular type of lesions, in addition Civatte bodies were identified in most samples of both clinical forms. For immunohistochemistry analysis, five fields with strong immunoreactivity for IL-17 and IL-23 were photomicrographed at 400x magnification, images were transferred to a computer where with ImageJ software®, lymphocytes that exhibited cytoplasmic immunostaining for these cytokines were counted. A mean was established after for each case. There was no statistically significant difference in the number of imunopositive lymphocytes for IL-17 and IL-23 among the group of OLP and IFH group, however a larger amount of lymphocytes imunopositive for IL-17 was found in the LPO group (p = 0.079) and significantly higher amounts of those lymphocytes were found in the erosive OLP when compared to the group of reticular OLP and IFH (p = 0.019). Furthermore, a marker epithelial immunopositivity for IL-17 was observed in OLP group. Although the results of this study do not permit the forceful assertion about the participation of Th17 lineage in OLP lesions, the findings of immunopositive lymphocytes counting for IL-17 and IL-23, which are potent proinflammatory cytokines, together with the the marked epithelial immunopositivity found for IL-17 in this study, suggest a possible role of this lineage in the pathogenesis of this disorder

Cistatina C e taxa de filtração glomerular em cirurgia cardíaca com circulação extracorpórea

OBJETIVO: Avaliar a cistatina C como marcador de função renal em pacientes submetidos à cirurgia de cardíaca com circulação extracorpórea, comparando com a dosagem sérica de creatinina. MÉTODOS: Foram analisados 50 pacientes consecutivos submetidos à cirurgia de revascularização do miocárdio. A função renal foi avaliada com a dosagem sérica de cistatina C e de creatinina no pré-operatório, no primeiro e no quinto dia de pós-operatório. Foram utilizadas as fórmulas de Cockcroft-Gault (CG) e Modification of Diet in Renal Disease (MDRD) para calcular a taxa de filtração glomerular estimada (TFG) através da creatinina, e a fórmula de Larsson para a TFG estimada através da cistatina C (TFG-Cis). RESULTADOS: A creatinina e o TFG através das fórmulas de CG e MDRD não mostraram diferença significativa nos momentos estudados. Após a agressão renal pela cirurgia, houve um aumento da cistatina C no 1º e 5º pós-operatório, sendo que no 5º pós-operatório com diferença estatisticamente significativa (P < 0,01). Houve uma queda da TFG estimada pela cistatina C de 105,2 ± 41,0 ml/min, no pré-operatório, para 89,5 ± 31,5 ml/min no 5º dia pós-operatório (P < 0,012). CONCLUSÃO: A cistatina C e a TFG-Cis apresentaram mudanças significativas no pós-operatório de cirurgia de revascularização do miocárdio quando comparadas a creatinina e a respectiva TFG estimada pelas fórmulas de Cockcroft-Gault e MDRD

Fatores pré-operatórios associados à injúria renal aguda após cirurgia cardíaca: estudo prospectivo

OBJECTIVES. To evaluate the clinical characteristics and co-morbidities associated with development of Acute kidney injury (AKI) after cardiac surgery in patients of a predominantly tertiary care university hospital.METHODS. One hundred and fifty cardiac surgery patients were prospectively studied at the Botucatu School of Medicine, University Hospital- UNEP, using a protocol which started 48h before and finished 48h after surgery, AKI was defined as a 30% increase in baseline serum creatinine.RESULTS. Were expressed as mean +/- standard deviation or median, with a significance level of 5%. Results: There was a slight male predominance (57%); mean age of the studied population, was 56 +/- 14 years. Sixty-five per cent of patients underwent myocardial revascularization. AKI was diagnosed in 34% of cases, Multivariate analysis showed that age over 60 years and peripheral vascular disease were significant associated with AKI developmentCONCLUSION. AKI was a common complication in cardiac surgery patients at this hospital, AKI was associated with age over 60 years and presence of peripheral vascular disease.

Fatores de risco para mortalidade na lesão renal aguda

OBJETIVO: Este trabalho tem como objetivo avaliar a evolução de pacientes com lesão renal aguda (LRA) por Necrose Tubular Aguda internados no Hospital das Clínicas da Faculdade de Medicina de Botucatu - UNESP. MÉTODOS: Trata-se de estudo de coorte retrospectivo, no qual foram avaliados 477 pacientes maiores de 18 anos, no período de janeiro de 2001 a dezembro de 2008. LRA foi definida de acordo com os valores de creatinina sérica, conforme proposto pelo Acute Kidney Injury Network (AKIN). RESULTADOS: A média de idade da população estudada foi de 65,5 ± 16,2 anos, com predomínio de homens (62%) e com idade > 60 anos (65,2%). Diabetes mellitus ocorreu em 61,9%, hipertensão arterial em 44,4% e doença renal crônica em 21,9%. A mortalidade foi de 66%. Após análise multivariada, foram variáveis associadas ao óbito a necessidade de diálise, internação em UTI, idade > 60 anos e menor tempo de acompanhamento nefrológico. A recuperação renal entre os sobreviventes foi de 96,9%. CONCLUSÃO: Este trabalho mostra que a evolução dos pacientes com LRA provenientes de enfermarias clínica e cirúrgica é semelhante à literatura. Porém, a alta mortalidade do grupo mostra a necessidade da identificação de fatores de risco para o desenvolvimento de LRA nesses pacientes e capacitação da equipe assistente para o diagnóstico precoce dessa síndrome.

Aspectos nutricionais na lesão renal aguda

A avaliação nutricional é ferramenta indispensável para a monitoração e acompanhamento clínico do paciente com lesão renal aguda (LRA). A perda aguda da função renal interfere no metabolismo de todos os macronutrientes, propiciando situações pró-inflamatórias, pró-oxidativas e de hipercatabolismo. As principais alterações nutricionais no paciente com LRA são hipercatabolismo, hiperglicemia e hipertrigliceridemia, que, somadas às contribuições da doença de base, complicações e necessidade de terapia renal substitutiva, podem interferir na depleção nutricional do paciente. A desnutrição em pacientes com LRA está associada a maior incidência de complicações, maior tempo de internação e maior mortalidade. Entretanto, existem poucos estudos na literatura avaliando o estado nutricional de pacientes com LRA. Parâmetros antropométricos como índice de massa corporal, circunferência do braço e pregas cutâneas são de difícil interpretação, devido à alteração no estado de hidratação desses pacientes. Os parâmetros bioquímicos geralmente utilizados na rotina clínica também sofrem influência de fatores não nutricionais, como prejuízo da função hepática e estado inflamatório. Embora não existam dados prospectivos sobre o comportamento dos marcadores nutricionais, alguns autores conseguiram demonstrar associações de alguns parâmetros com desfecho clínico. A utilização de marcadores como albumina, colesterol, pré-albumina, IGF-1, aplicação da avaliação subjetiva global e cálculo do balanço nitrogenado parecem ser úteis como parâmetros de triagem para pior prognóstico e maior mortalidade em pacientes com LRA. em pacientes com LRA em terapia renal substitutiva, uma oferta calórica em torno de 25 a 30 kcal/kg e oferta mínima de 1,5 g/kg/dia de proteínas é recomendada a fim de minimizar o catabolismo proteico e prevenir complicações metabólicas.

Análise imuno-histoquímica de proteínas relacionadas às respostas Th1, Th2 e Th17 na doença periodontal

Periodontal disease is an inflammatory condition of infectious nature characterized by destruction of protecting and supporting dental tissues. It happens as a response produced by the host when attacked by microorganisms. Several factors are involved in this process. Among them, cytokines are key regulatory molecules in this immune response, playing a role either protective and/or destructive in lesion progression. Thus, this study investigated the immunohistochemical expression of IFN- , GATA-3, IL-17, IL-23, IL-6 and TGF- in gingival tissues of humans, in an attempt to gain a better understanding of the participation of Th1, Th2 and Th17 immune responses in the development of periodontal disease processes. To this end, eighty-two samples of gingival tissues were divided into three groups: Group 1 = 15 (samples of healthy gum tissue as controls), Group 2 = 36 (samples with chronic gingivitis) and Group 3 = 31 (samples with chronic periodontitis). All cases were submitted to morphological analysis from sections stained with hematoxylin and eosin and then subjected to staining by immunohistochemistry using the streptavidin-biotin method. Results showed positive labeling for all proteins. Nonetheless, we observed a greater expression of Th1 cytokines and Th17 cells in group 3. We found statistically significant difference between TGF- expression and the clinical condition of the samples (p=0.02). We conclude that Th1 and Th17 responses may act synergistically in the destructive process of periodontal tissue, overlapping the Th2 response that was also present in these tissues