Design, Synthesis, and Pharmacological Evaluation of Novel Hybrid Compounds To Treat Sickle Cell Disease Symptoms

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

Cocaine induces cell death and activates the transcription nuclear factor kappa-b in pc12 cells

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

Experimental murine mycobacteriosis: evaluation of the functional activity of alveolar macrophages in thalidomide- treated mice

Thalidomide is a selective inhibitor of tumor necrosis factor-alpha (TNF-alpha), a cytokine involved in mycobacterial death mechanisms. We investigated the role of this drug in the functional activity of alveolar macrophages in the presence of infection induced by intranasal inoculation of Mycobacterium avium in thalidomide-treated and untreated adult Swiss mice. Sixty animals were inoculated with 5 x 10(6) M. avium by the respiratory route. Thirty animals received daily thalidomide (30 mg/kg mouse) and 30 received water by gavage up to sacrifice. Ten non-inoculated mice were used as a control group. Lots of animals from each group were evaluated until 6 weeks after inoculation. Infection resulted in an increased total number of inflammatory cells as well as increased activity of pulmonary macrophages. Histologically, intranasal inoculation of bacilli resulted in small mononuclear infiltrates located at the periphery of the organ. Culture of lung fragments revealed the presence of bacilli only at the beginning and at the end of the experimental period. Thalidomide administration did not affect the microbiological or histological features of the infection. Thalidomide-treated and untreated animals showed the same amount of M. avium colonies 3 weeks after infection. Although it did not affect bacillary clearance, thalidomide administration resulted in a decreased percent of spread cells and release of hydrogen peroxide, suggesting that factors other than TNF-alpha play a role in the killing of mycobacteria by alveolar macrophages. Thalidomide administration also reduced the number of spread cells among resident macrophages, suggesting a direct effect of the drug on this phenomenon.

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.

4-Fluoro-2-methoxyphenol, an apocynin analog with enhanced inhibitory effect on leukocyte oxidant production and phagocytosis

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

Expressão imuno-histoquímica do cd8, foxp3, Tgf β, tnf α e nf-кb em displasias epiteliais e Carcinomas epidermóides orais

The Oral Epithelial Dysplasia (OED) is the lesion that precedes or co-exists with the Oral Squamous Cell Carcinoma (OSCC), presenting molecular and/or histological similar alterations. The divergences about the malignization potential of OEDs and the role of inflammation in this process make hard the early diagnosis and evaluation of OSCCs aggressiveness. Thus, it became the goal of this study to evaluate the role of inflammation in oral carcinogenesis and tumoral aggressiveness. For this purpose a morphological study was performed in 20 OED cases and 40 OSCC cases to detect the malignization potential of OEDs and the histologic malignancy grading (HMG) of OSCCs, analyzing superficial masses for dismorphism evaluation and the invasive front for evaluation of tumoral growing; and immunohistochemical, using anti-CD8, anti-FOXP3, anti-TGFβ, anti-TNFα and anti-NF-кB antibodies, comparing their with the types lesion, histological degree and intensity of the inflammatory infiltrate. The results were statistically significant for the parameters: cell maturity (p=0,0001), masses presence (p=0,038) and dismorphism (p=0,037), when associated to HMG. To compare the expression of the markers with the types lesion, a significantly higher expression of CD8 (p=0,001) and NF-кB (p=0,002) in the OED, and also a smaller expression of the epithelial TGFβ in the severe OEDs (p=0,011), without significant expression between OSCC degrees. By relating the expression of the studied markers with the inflammatory infiltrate intensity, a positive relation was observed with: inflammatory TNFα(p=0,003), epithelial TNFα and NF-кB (p=0,051 and p=0,004), in OEDs; and with CD8 (p=0,021) and TNFα (p=0,015) in conjunctive OSCCs; and a negative relation with epithelial TNFα (p=0,034) in OSCCs. No significant relation was found between FOXP3 with any of the studied variables. These findings lead to the conclusion that, the study of the invasive front is as important as the study of superficial masses for the evaluation of tumoral aggressiveness; the intensity of the inflammatory infiltrate has no use as a parameter for prognostic evaluation of OSCC in routine exams, but, the molecular events detected in this study may be necessary to give basis for determining the malignant potential in OEDs and aggressiveness in OSCCs

Adaptação e validação da escala de motivação à prática de atividades físicas

The study of the motivation to practice physical activities is relevant once through this it is possible to develop intervention strategies for sedentary population. In this way, the present dissertation aimed to adapt and validate for the Brazilian context the Motives for Physical Activity Measure Revised (MPAM-R). It was investigated yet the relation of some socio-demographic variables (age, gender, Body Mass Index - BMI, types of physical activity; way to practice and time of practice) and the participants means in the motives studied. The physical activity (PA) is defined as any bodily movement produced by skeletal muscles resulting in energy expenditure above the basal level. However, in the present study it was considered motivation for two types of PA: the physical exercises and sports. The Self-Determination Theory (SDT) underlay this research, once it has been used in the sportive context, beyond the facto of it was used as theoretical base to the development of MPAM-R. To attain the proposed goals, it was accomplished translations to Portuguese of the original English scale. Next, was carried out the semantic analysis and the judge analysis. For the empirical analysis of items, participated 309 practitioner of PA, classified in physical exercises practitioners and sports practitioners, with ages between 16 and 74 years, distributed equally by sex. They answered the final version in Portuguese of the MPAM-R and socio-demographic questions. The data was collected in Natal/RN, where the researcher approached people in some places where the PA is practiced, following she communicated that their involvement will be spontaneous and their responses will be confidential. The obtained results pointed the confirmation of the existence of five factors in the final version of the instrument, that was composed by 26 items that presented the following statistics indexes: x² (289) = 757.75, p < 0.000, x²/DF = 2.62, with GFI of 0.83, AGFI of 0.80 and RMSEA of 0.07. The reliability (Cronbach Alpha) of the complete instrument was 0.90, and the indexes of each factor were considered satisfactory too: Enjoyment (α = 0.88), Health (α = 0.84), Appearance (α = 0.79), Competence (α = 0.85) and Social (α = 0.75). It was observed that, in general, the main motive presented by the participants to practice PA was Health. It was verified yet, that women and aged had a higher mean in the Health factor; among the exercise practitioners was found a higher mean in the Appearance factor; and a higher mean in the Social factor was found among those that practice PA with accompaniment. It was concluded that the MPAM-R presented satisfactory psychometric parameters, became it useful in futures researches. Moreover, proposed the accomplishment of new studies that considered others variables to the intent of the better understanding of motivation to practice physical activity

Adaptação e validação da escala de dependência de exercício físico em versão tradicional e informatizada

While providing physical and psychological benefits, excessive exercise could be or cause a compulsive behavior, making the individual dependent on it. In a parallel discussion, computerized psychological instruments, for a hand, reflects the development of information technology and your applicability to other areas, but also shows little advance for Psychological Assessment. In this perspective, this study aims to adapt the Exercise Dependence Scale (EDS-R) in two formats (paper-and-pencil and computerized) and evaluate evidence of factorial and convergent validity, and reliability of each version and compare them with each other. It is also proposed to observe the relationship of some bio-demographic (Sex, age, frequency, duration and intensity of practice exercise) and the exercise dependence (DEF). For this purpose, 709 regular physical activity practitioners, selected by procedures non-probabilistic sampling, responded a adapted version of EDS-R, Muscle Appearance Satisfaction Scale (MASS), Body Modification Scale (BMS) and a demographic questionnaire, analyzed through Exploratory Factor Analysis, Cronbach's Alpha and not parametric tests. Both the traditional version and the computer showed a seven factors structure, explaining 57 and 62% of the variance, respectively, and Cronbach's alphas of 0.83 and 0.89. Factors were: (1) intentionality, (2) continuity, (3) tolerance, (4) reduction of other activities, (5) lack of control, (6) abstinence and (7) time spent on exercise. Relationships were observed between the Exercise Dependence and the variables: age, diets, consumption of food supplements and medicines for weight change, desire to do plastic surgery and body satisfaction. We observed also a positive correlation between the DEF and the frequency, duration and intensity of exercise, and the factor "Dependence on exercising" from MASS, indicating convergent validity of the EDS-R. Finally, comparisons between the two formats were equivalent, with few changes: computerized version achieved higher DEF scores. Based on these results, it can be concluded that the EDS-R has factorial and convergent validity, reliability, to measure exerceise dependence on traditional e computerized formats. DEF is related to actions used to body modification and behaviors toward exercise. Finally, it was found equivalence between the formats, especially in psychometric parameters, thus suggesting feasibility of a computerized assessment. However, it was observed that the computerized data has sample recruiting strategies more limited

Tradução e adaptação da escala de dependência de substâncias do Millon Clinical Multiaxial III para o Brasil

Millon describes the normal personality by means of adaptation styles that are effective in normal environments and personality disorders such as unadapted operating styles. To operacionalize his theoretical model, Millon has built several instruments, including the Millon Clinical Multiaxial Inventory III (MCMI-III), wich consists of a self report inventory composed by 175 true or false response items, containing four verification scales, and others scales wich evaluates 14 personality patterns and 10 clinical syndromes. The Substance Dependence scale (T) is placed along with Clinical Syndromes scales. This research is justified by the lack of a Brazilian instrument to assess personality psychopathological aspects, and aims to translate and semantically adapt the MCMI-III to the Brazilian context, checking validity elements of the Substance Dependence scale, and developing a computer application for assisting the evaluation of assessment results. To this intent, 2.588 individuals data was collected, male and female, aged between 18 and 85 years, characterized as belonging to a clinical or non-clinical group, who took part in the survey via the internet or in person. Respondents completed the MCMI-III, a socio-demographic questionnaire and a subgroup also answered to the Goldberg General Health Questionnaire (GHQ). Besides descriptive statistics, we performed the analysis using the Student t test, principal components analysis and internal consistency. Despite difficulties related to translating very specific English terms, the assessment by judges, experts on Millon´s theory, and the back translation, attested the adequacy of the Brazilian version. Factorial analysis indicated the grouping of translated T scale items into three factors (social activities prejudice, lack of impulse control, and oppositional behavior), by presenting a single item on a fourth factor (apparently related to seeking pleasurable stimuli). The Cronbach alpha for this set of items was 0,82, indicating an acceptable scale reliability. The data analysis resulted in distinction of scores between clinical and non-clinical groups and between men and women; the relationship between high scores on the scale T and the other scales; scores of drug users according to the declared used substance; and the relationship between high scores on T and the verification of disorder or risk on GHQ mental health factor, indicating the instrument´s adequate sensistivity in identifying psychopathologies and the relationship between the different disorders or psychopathological personality patterns. Although further studies are necessary to develop the scores transformation factors, the computerized correction tool was adequate.

TLR4 and CD14 receptors expressed in rat pineal gland trigger NFKB pathway

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

Infliximab-induced psoriasis during therapy for Crohn's disease

Although therapy with tumor necrosis factor-alpha inhibitors (anti-TNF) provides beneficial effects in different immune inflammatory disorders, paradoxical cases of anti-THE-induced psoriasis have increasingly been reported, mostly in the setting of rheumatologic diseases. To date, less than 50 cases of infliximab-induced psoriasis in inflammatory bowel disease patients have been described. The present report was aimed at describing two new cases of infliximab-induced psoriasis during therapy for Crohn's disease and at carrying out a review on this intriguing phenomenon. (C) 2011 European Crohn's and Colitis Organisation. Published by Elsevier B.V. All rights reserved.

Cytokines released from blood monocytes and expressed in mucocutaneous lesions of patients with paracoccidioidomycosis evaluated before and during trimethoprim-sulfamethoxazole treatment

Background Mucocutaneous lesions in paracoccidioidomycosis are granulomatous and result from tissue responses to Paracoccidioides brasiliensis, the aetiological agent.Objectives and methods In this study we investigate the expression of tumour necrosis factor (TNF)-alpha, interleukin (IL)-10 and transforming growth factor (TGF)-beta 1 by immunohistochemistry in skin and mucosa lesions from patients with the chronic form of paracoccidioidomycosis, evaluated before and at day 20 of trimethoprim-sulfamethoxazole treatment. Cytokine production by peripheral blood monocytes was also studied by enzyme immunoassay.Results Intense immunostaining for TNF-alpha was detected in mononuclear cells that infiltrated granulomas in all skin and mucosa lesions before treatment simultaneously with low IL-10 granular deposits in these cells. At day 20 of treatment, there was reduced TNF-alpha and IL-10 deposition. Immunoreactive TGF-beta 1 was observed diffusely in the dermis and generally in the cytoplasm of macrophages and giant cells, before treatment, and as increased TGF-beta 1 deposits in the fibrosis area at day 20 of treatment. Peripheral blood monocytes from patients with paracoccidioidomycosis, evaluated before treatment, produced high endogenous levels of TNF-alpha, TGF-beta 1 and IL-10 in relation to healthy controls. Lipopolysaccharide-stimulated monocytes from patients secreted lower levels of TNF-alpha in both periods of evaluation while no impairment in capacity of IL-10 and TGF-beta production was observed.Conclusions Trimethoprim-sulfamethoxazole therapy was effective in decreasing fungal load in the lesions, allowing patient immune response to control the infection leading to the healing of the lesions.

HISTORICAL PERSPECTIVE and HUMAN CONSEQUENCES of AFRICANIZED BEE STINGS IN THE AMERICAS

In 1956, Africanized bees began to spread in the American continent from southern Brazil, where original African bees mated with European bees. A few years later, in 1990, these Africanized bees reached the United States and were found in Texas. Currently, these hybrid bees are found in several North American states and will probably reach the Canadian border in the future. Although the presence of Africanized bees had produced positive effects on Brazilian economy, including improvement in crop pollination and in honey production, turning Brazil into a major exporter, the negative impacts-such as swarming, aggressive behavior, and the ability to mass attack-resulted in serious and fatal envenomation with humans and animals. Victims of bee attacks usually develop a severe envenomation syndrome characterized by the release of a large amount of cytokines [interleukins (IL) IL-1, IL-6, IL-8], and tumor necrosis factor (TNF). Subsequently, such cytokines produce an acute inflammatory response that triggers adverse effects on skeletal muscles; bone marrow; hepatic and renal functions; and cardiovascular, central nervous, and immune systems. Finally, the aim of the present review is to study historical characteristics and current status of Africanized bees' spread, the composition of their venom, the impact of the bees on the Brazilian economy and ecology, and clinical aspects of their stings including immune response, and to suggest a protocol for bee sting management since there is no safe and effective antivenom available.

Modulatory effect of prostaglandins on human monocyte activation for killing of high- and low-virulence strains of Paracoccidioides brasiliensis

The effect of indomethacin (Indo), a cyclo-oxygenase inhibitor, on the monocyte-mediated killing of a low-(Pb265) and a high-(Pb18) virulence strain of Paracoccidioides brasiliensis was examined. The Pb18 strain was not killed by either non-activated or interferon-gamma (IFN-gamma)-activated human monocytes but these cells did show fungicidal activity if pretreated with Indo. In contrast with IFN-gamma tumour necrosis factor-alpha (TNF-alpha) was very effective at stimulating the fungicidal activity of monocytes. While the low-virulence strain, Pb265, could not be killed by monocytes, cells preincubated with IFN-gamma demonstrated fungicidal activity. The killing of this strain was also induced by pretreatment of monocytes with Indo. The results suggest a negative role for prostaglandins, which are synthesized via the cyclo-oxygenase pathway, in the regulation of monocyte-mediated killing of virulent and avirulent strains of P. brasiliensis and that TNF-alpha generation during the fungus-monocyte interaction is more important in the killing of Pb265 than Pb18.

Granulocyte macrophage colony-stimulating factor enhances the modulatory effect of cytokines on monocyte-derived multinucleated giant cell formation and fungicidal activity against Paracoccidioides brasiliensis

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