Indução de morte celular em células de adenocarcinoma cervical humano (HeLa) pela lectina da esponja Cinachyrella apion (CaL)

Cancer is a term used to represent a set of more than 100 diseases, including malignant tumors from different locations. The malignancies are the second leading cause of death in the population, representing approximately 17% of deaths of known cause. Strategies that induce differentiation have had limited success in the treatment of established cancers. In this work, a lectin purified from the marine sponge Cinachyrella apion (CaL) was evaluated due to its hemolytic, cytotoxic and antiproliferative properties, besides the ability to induce cell death via apoptosis in tumor cells. The antiproliferative activity of CaL was tested against cell lines, with the highest inhibition of tumor growth for HeLa, reducing cell growth at a dose dependent manner, with a concentration of 10 μg/mL. The hemolytic activity and toxicity against peripheral blood cells were tested using the concentration of IC50 for both trials and twice the IC50 for analysis in flow cytometry, indicating that CaL is not toxic to these cells. To assess the mechanism of cell death caused by CaL in HeLa cells, we performed flow cytometry and western blotting. The results showed the lectin probably induces cell death by apoptosis activation by pro-apoptotic protein Bax, promoting mitochondrial membrane permeabilization, cell cycle arrest in S phase, with accumulation of cells of approximately 57% in this phase, and acting as both dependent and/or independent of caspases pathway. These results suggest that CaL has the potential to be used as drug treatment against cancer.

O papel modulatório do ferro sobre a resposta imune na Leishmaniose Visceral

Iron is an essential element for many cellular functions, including the immune response against intracellular pathogens. In this study, we aimed evaluate the effect of iron on IRP2, IFN-γ, TNF-α, IL-6, IL-10, MIG and IP10 expression in PBMC and assess the effect of the spleen parasite load on the expression of these genes in the spleen of L. infantum naturally infected dogs. Blood sample from 7 DTH+ donor was collected and PBMC was obtained. The cells were cultivated in absence (iron chelator desferroximane, DFO 10 μM supplemented media) or in presence of iron (hemin 6 mM) for 1 h, followed by stimulation with Leishmania infatum antigen for 4 h. 44 dog spleen samples were obtained and parasite load in this organ was determinate by qPCR. Gene expression was analyzed by qPCR and cytokine production quantified by flow cytometry. In antigen stimulated cells, genes involved in immune response are significantly more expressed in presence of iron. T CD4+ and TCD8+ lymphocytes produces IFN-γ, TNF-α and IL-10 possibly in iron dependent pathway. Monocytes antigen stimulated reduced TNF-α, IL-6 and IL-10 production in presence of iron. We found spleen of infected dogs IRP2 expression increases according to parasite load in that organ, while an inverse profile was found for IFN-γ, TNF-α e IL-10 expression. These results suggest that T lymphocytes depends on iron to produce IFN-γ, TNF-α and IL-10, while iron seems to inhibit cytokine production in monocytes. So, we propose an immunoregulatory mechanism carried out by iron during L. infantum infection in humans and dogs

Avaliação do efeito da megadose de vitamina A no colostro humano

The mothers supplementation of vitamin A in the postpartum comes being a measure of intervention sufficiently used in the combat to the vitamin deficiency. The objective of this work was to evaluate the effect of the mother megadose of vitamin A under the levels of retinol in colostrum of postpartum mothers receiving care at the Januário Cicco Maternity School (MEJC), Natal, RN, as well as analyzing the influence of the maternal nutritional status in the reply to this supplementation. The study it was transversal type, with participation of 91 women in labor divided in group had participated of the study have controlled (44 women) and supplemented group (47 women). In the period of the morning blood and milk had been collected (milk 0h). After that a capsule of retinil palmitate of (200 000 UI or 60 mg) was supplied to the supplemented group. Another aliquot of colostro was after gotten 24h of the first collection (milk 24h). Retinol in milk and serum was quantified through the High Pressure Liquid Chromatography. The vitamin ingestion was evaluated by the questionnaire of frequency of alimentary consumption. The levels of serum retinol were 40.6 ± 10.6 and 35.9 ± 10.9 µg/dL in the groups controlled and supplemented, respectively. The women had presented a satisfactory average ingestion of vitamin (1492,4 µgRAE/dia), however with high prevalence of inadequate consumption (23%). Average values of retinol in milk 0h had been found and 24h of 93.5 ± 50.3 µg/dL and 99.1 ± 49.3 µg/dL has the group controlled group, respectively (p>0.05). After the supplementation had a significant increase in the levels of retinol of the supplemented group, being found values of 102.0 ± 56.0 µg/dL and 196.1 ± 74.0 µg/dL for milk 0h and 24h, respectively (p<0.0001). The women in labor presented different answers to the supplementation influenced for the basal levels of retinol in colostrum. It was possible to verify that women with deficient levels of retinol in milk had transferred more retinol to milk 24h than ones with adjusted levels, showing a percentage of reply equivalent to 326.1% and 86.5% of increase, respectively (p< 0.0001). Although the apparent normality found in the serum, the studied women are considered of risk to the development of the vitamin deficiency, and megadose was efficient in first 24h after the supplementation and wakes up with the mechanisms considered for transference of vitamin A to the milk

Prevalência da infecção por papilomavírus humano, herpes simplex tipo 1 e 2 e Chlamydia trachomatis em um segmento da população feminina da grande Natal/RN

Sexually transmitted diseases (STDs) are among the largest public health problems, especially in developing countries. The acquisition of these infections during early sexual activity is common and many infections have a benign course. However, in some pathogens remain in the state of latency can be reactivated and cause productive infection that may progress to severe forms. In addition, some of them are transmitted vertically resulting in congenital infection, causing immediate damage or long-term child. The classic risk factors for sexually transmitted agents are: early onset of sexual and reproductive health, multiple sexual partners throughout life, use of oral contraceptives and co-infections with different pathogens. We present the results of a cross-sectional study aimed to estimate the prevalence of genital infection by human papillomavirus (HPV), Herpes simplex virus (HSV) and Chlamydia trachomatis (CT) in a segment of the female population of the metropolitan area Christmas, among those who enrolled voluntarily sought, Basic Health Units for the examination of cancer screening cervix in the period 2008 to 2010. All participants, a total of 261 women answered a standard questionnaire by which identified the socio-demographic characteristics, classical risk factors for STDs, reproductive and sexual activity and smoking. Of each patient were obtained two samples, one for the completion of the Pap test for detection of cellular changes and the other processed for DNA extraction and analyzed by PCR (polymerase chain reaction) to detect the three pathogens studied. The population of the study was composed of sexually active women aged between 13 and 79 years, mean 38.7 years, most of them being married, low education levels and low incomes. The majority (87%) had normal results on cytology and only 2.7% had low-grade cytological abnormalities. Prevalence rates were 37.9% for HPV, 4.6% for CT and 26% for HSV. HPV prevalence was higher in women under 25, unmarried and in those who had multiple sexual partners. Women with simultaneous infection by HSV-1 and 2 had higher prevalence of HPV infection. The prevalence of HSV infection showed no association whatsoever with the risk factors analyzed and HSV-1 was the predominant type among the cases of genital HSV infection. The overall prevalence of C. Trachomatis was relatively low, thus providing greater value in younger women aged less than or equal to 20 years

Prevalência da infecção pelo Papilomavírus humano, Chlamydia Trachomatis e Herpes Simples do tipo 2 em adolescentes atendidas em unidades de saúde pública de Natal

This study assessed the level of knowledge, attitude and practice of Pap smear and human papillomavirus (HPV), in addition to analyzing the prevalence of genital HPV infection, Herpes Simplex Type 2 (HSV-2) and Chlamydia trachomatis in teenagers. The study consisted of two approaches, one based only on interviews conducted with adolescents enrolled in public schools or in public health facilities in the city of Natal. The other approach involved only a group of 132 adolescents enrolled among those admitted to two health units in Natal-RN. This second group of participants two specimens were collected for laboratory analysis: one was directed to prepare the blade for the Pap test, and other processed for DNA extraction for molecular analysis, focusing on the detection of HPV, HSV-2 and C . trachomatis. The presence of DNA of the three pathogens was investigated by the technique of polymerase chain reaction (PCR). The presence of each of the three pathogens was analyzed in terms of socio-demographic characteristics, as well as sexual and reproductive activity to identify risk factors for infection and development of lesions of the uterine cervix. The results show that the adolescents in this study had levels of knowledge and attitude very low, both in relation to cytology to HPV as though they have made a reasonable percentage of adequate practice exam and prevention of HPV infection. The overall prevalence of HPV infection was 54.5% and 48.2% in adolescents with normal cytology and 86.4% in those with abnormal cytology. We observed a higher proportion of cases of infection in the age group of 18 to 21. The prevalence of HPV infection was slightly higher among pregnant teenagers. The overall prevalence of HSV-2 infection was 13.6% and 11.8% in women with normal cytology and 22.7% in those with abnormal cytology. A higher proportion of cases of infection was found in the age group from 14 to 17, with a slightly higher prevalence among pregnant women. The C. trachomatis was found with an overall prevalence of 19.7% and 21.8% in adolescents with normal cytology and 9.1% in those with abnormal cytology. The prevailing rate was highest in the age group 18 to 21 years and in nonpregnant

Correlação entre a infecção genital pelo vírus do papiloma humano, a resposta imune e os achados colpocitológicos em mulheres grávidas e não grávidas

The genital HPV infection is very common between men and women worldwide, affecting particularly young women, constituting a serious public health problem in less developed regions, favored by the poor living conditions of population. The cytology and colposcopy have notorious importance in the diagnosis of precursor lesions of cervical cancer and therefore its prevention. However, even with such diagnostic tools, the number of women who develop cervical cancer is still high. This study aims to assess the prevalence of genital tract infection by HPV in pregnant and nonpregnant women, evaluating the profile of the immune response presented by the women of these two groups in order to establish correlations among profile of immune response, presence of virus and occurrence of lesions of the uterine cervix. We analyzed specimens obtained from the cervix of 221 patients, 91 pregnant and 130 non-pregnant, aged 14-72 years. The women were subjected to colposcopic and cytologic evaluation detect possible changes in the cervix and then samples were collected in order to perform HPV detection by PCR and real-time PCR for detection of mRNA of pro-inflammatory and anti-inflammatory cytokines. In the present study, the overall prevalence of HPV genital infection was 28.1%; of which 31.9% were pregnant patients and 25.4% in non-pregnant women. Young women under 30 years and those with low educational level education showed a higher risk of HPV infection. Colposcopy showed better correlation with detection of HPV DNA by PCR, when compared to cytology. Generally, HPV infected patients, pregnant or not, exhibited reduced mRNA expression of both pro-inflammatory (IFN-γ, TNF-α) and anti-inflammatory (IL -10) cytokines, when compared to patients not infected by HPV. Nonpregnant patients infected presented increase mRNA expression of IL-17 in patients without injury, whereas those with lesion showed higher mRNA expression of TGF-β. Pregnant women without injury infected exhibited increased mRNA expression of TGF-β. There was no difference in HPV prevalence between pregnant and nonpregnant women. There was a reduction of pro-inflammatory cytokines, except IL-17, in all women infected by HPV. Moreover, we observed an increase of TGF-β in HPV-infected women who are pregnant or not. The results suggest that, in women in this study, HPV infection promoted changes in the profile of cytokines necessary for activation of effective immune response, possibly favoring viral persistence

Estudo in silico da interação da albumina de soro humano com o ibuprofeno

Currently, computational methods have been increasingly used to aid in the characterization of molecular biological systems, especially when they relevant to human health. Ibuprofen is a nonsteroidal antiinflammatory or broadband use in the clinic. Once in the bloodstream, most of ibuprofen is linked to human serum albumin, the major protein of blood plasma, decreasing its bioavailability and requiring larger doses to produce its antiinflamatory action. This study aimes to characterize, through the interaction energy, how is the binding of ibuprofen to albumin and to establish what are the main amino acids and molecular interactions involved in the process. For this purpouse, it was conducted an in silico study, by using quantum mechanical calculations based on Density Functional Theory (DFT), with Generalized Gradient approximation (GGA) to describe the effects of exchange and correlation. The interaction energy of each amino acid belonging to the binding site to the ligand was calculated the using the method of molecular fragmentation with conjugated caps (MFCC). Besides energy, we calculated the distances, types of molecular interactions and atomic groups involved. The theoretical models used were satisfactory and show a more accurate description when the dielectric constant ε = 40 was used. The findings corroborate the literature in which the Sudlow site I (I-FA3) is the primary binding site and the site I-FA6 as secondary site. However, it differs in identifying the most important amino acids, which by interaction energy, in order of decreasing energy, are: Arg410, Lys414, Ser 489, Leu453 and Tyr411 to the I-Site FA3 and Leu481, Ser480, Lys351, Val482 and Arg209 to the site I-FA6. The quantification of interaction energy and description of the most important amino acids opens new avenues for studies aiming at manipulating the structure of ibuprofen, in order to decrease its interaction with albumin, and consequently increase its distribution

Hospital humano: etnografia da humanização hospitalar na perspectiva de usuários

This ethnographic work studies the experiences of patients admitted in public (PUH) and private (PRH) hospitals in the Brazilian northeastern region. 28 adult patients of different clinics participated in the study. Data were analyzed by the patient path method, consisting in a combination of complemented and articulated techniques free observation, participating observation, ethnographic interview and patient testimonials collected prospectively during the patients admissions, from their arrival and until their discharge. The analysis was carried out according to the Thematic Categories Analysis Technique and the data were interpreted pursuant to medical anthropology, healthcare humanization and healthcare promotion theoretical references. The ethical principles of Resolution 196/96 were followed. The human hospital, as revealed by the patient, highlights the significance of subjectivity. 225 (54.7%) out of 411 mentioned concepts were collected in a public hospital (PUH) and 186 (45.3%) in a private institution (PRH). The results show that the patient at the PUH and PRH ethnoevaluates different aspects of the healthcare professionals´ human and technical competence, the hospital´s functioning structure, the access to and the ethics in the financial management, and develops overcoming strategies for his stay at the hospital. This ethnoevaluation is mediated by different factors, namely: social and economic status, personality, religiosity, ironic speech, somber diagnosis and satisfied needs, prior hospital experiences and the conditions under which the interview was carried out. A pedagogic proposal for the hospital humanization must include structural, managerial and organizational changes of the offered services and use active methodologies aimed to the political resolution of problematic situations at work and the inclusion of affective and subjective factors, and become as well a tool for the collective learning. This study shows the importance for the user´s ethnoevaluation to be incorporated into the hospital management and care as a guideline in the decision making and clinical action, thus promoting practices that shall lead to a decent and humanized care. The multidisciplinary nature of this study allowed a wide understanding of the user´s perspective as a socially critical ethnoevaluator

Caracterização do perfil imunológico em mulheres portadoras do papilomavírus humano

O câncer de colo uterino consiste em um problema de grande relevância social, visto que se trata da segunda malignidade ginecológica mais freqüente no mundo. O principal precursor do câncer da cérvice uterina consiste na infecção pelo Papiloma vírus humano (HPV). O HPV tem despertado grande interesse na comunidade científica, devido à relação estabelecida entre a sua epidemiologia e o câncer de cérvice uterina. Apesar do grande avanço na biologia do HPV, pouco se sabe acerca da resposta imunológica a este vírus. O Papilomavírus humano (HPV) é um DNA vírus epiteliotrófico, que está vinculado à carcinogênese do colo uterino por meio de evidências epidemiológicas e laboratoriais. As infecções pelo HPV ocorrem em mulheres em todo o mundo. Existe consenso na literatura quanto à associação do DNA HPV com as neoplasias intra epiteliais cervicais e o câncer cervical. Por este motivo é de grande importância a detecção precoce deste tipo de infecção para que seja efetuado um tratamento específico. Esse trabalho está caracterizando o perfil imunológico de pacientes portadoras de HPV através da quantificação linfocitária (TCD4, TCD8, células NKT e células NK), e correlacionou esse material à técnica de captura híbrida, para poder comprovar a infecção pelo HPV, esperando assim contribuir precocemente para o diagnóstico do câncer de colo uterino.

Liofilização como ferramenta de preservação de sêmen humano

Liofilização é um método alternativo de preservação de espermatozóide, com vantagens sobre o criopreservado pelo baixo custo de armazenamento, facilidade no transporte pela não necessidade de containers e baixas taxas de contaminação. Esse estudo foi realizado no intuito e testar a eficácia de 5 açúcares na proteção do material cromossômico (DNA) de espermatozoides humanos liofilizados após reidratação. Para isso foram utilizados espermatozoides de indivíduos classificados como normozooespérmicos segundo OMS. Em eppendorfs foram colocados 1ml de sêmen que foram diluídos (1:1) em 1 ml de solução de Glicose, Lactose, Maltose, Manitol e Sorbitol, separadamente, nas concentrações 0.2M, 0.4M, 0.5M e 0.6M. Essas amostras foram analisadas em duas fases a primeira após congelamento observando a porcentagem de células com danos de membrana e outra análise após a liofilização, utilizando o Teste Cometa para detectar as taxas de fragmentação de DNA dos espermatozoides recuperadas após reidratação. O processo de liofilização de espermatozoides não foi capaz de recuperar a motilidade dos mesmos, mas foi capaz de manter 62.30 ± 13.76% de membrana plasmática intacta na presença de 0.6M manitol. As substâncias que melhor preservaram o DNA dessas células foram 0.2 lactose, 0.2M e 0.6M manitol e sorbitol, com apenas 4% de danos ao DNA. Esses resultados sugerem uma boa proteção ao DNA de espermatozoides humanos liofilizados, mas ainda requer testes adicionais para avaliar a fertilização e estudos que garantam a viabilidade da prole

IgA sérica, secretora, IgE total e estado nutricional em crianças com infecções por enteroparasitas

Infections for intestinal parasites are one of the main morbidade causes in humans and, its relationships with socioeconomic levels and hygiene conditions in countries in development are already very established. Many works, even so, they are being accomplished to elucidate the complex interactions among nutrition, these infections and answer imunológica, because it is seen that malnutrition commits the immunity increasing the susceptibilidade for infectious diseases and these for its time can harm the state human nutricional. It is known that sponge helmínticos they stimulate synthesis of IgE so much policlonal as specific for the same ones and that IgA secretora, main imunoglobulina of defense of the mucous ones, can act against protozoa as the Giardia lamblia and against helmintos as Trichuris tichiura and Strongyloides stercorales. Some studies show that the malnutrition energy protéica influences in the production of these answers, but some authors show results divergentes. In this work it was evaluated the levels of total IgE, IgA sérica and secretora, contagem of sanguine eosinófilos, levels of proteins séricas and state nutricional, in 103 children of low socioeconomic level, to discover a correlation between those and infection for enteroparasitas. They participated in the study children of both sexes, with age of 3 to 6 years, visitors of the same creche and residents in a neighborhood with precarious hygiene conditions and basic saneamento, in the city of Christmas. The obtained results showed that the faulty environmental and socioeconomic conditions favored to a high infection frequency for enteroparasitas, mainly Trichuris trichiura and Ascaris lumbricoides between the helmintos and Endolimax sleep and Giárdia lamblia among the protozoa. Light malnutrition without deficit protéico was observed in 30% of the children, which didn't also present significant deficiencies of IgA sérica and secretora. The sponged children

Protagonismo juvenil: uma avaliação do programa agente jovem de desenvolvimento humano na cidade de Natal RN (2006/2007).

This paper presents a critical rescue of the term youth, while simultaneously is presented as the lifetime of between 15 and 24 years. Rescue two features in the literature. The first considers youth as a transition period and the second phase work that from a predisposition to rebellion. Discusses the design plural of that term, youth recital to this social and historical aspects of different societies to which it relates and highlights the importance of realizing the diversity of it. Where the object of investigation, the Young Agent Program, the overall objective of the study is to evaluate the effectiveness of the actions of the Young Agent Program for Human Development in Natal-RN. Specifically aims to evaluate the effectiveness of the Young Agent Program: a) as an enhancer of the integration of young people in social spaces of family, school and community, b) as to (re) insertion and the incorporation of issues related to health in adolescence, and c ) regarding the work of young people as multipliers of the actions of the program. With regard to the methodological procedures presents a review of the literature on youth, and the categories of human development, leadership and evaluation of youth policies for the construction of theoretical and analytical approach, using documentary data collection in the Municipal Labor and Social Services - SEMT on the Young Agent Program, and interviews with actors involved in the program. Finally, evaluates the effectiveness of the Young Agent Program from the perspective of participants

Água de consumo humano como fator de risco à saúde em propriedades rurais

OBJETIVO: Verificar a qualidade higiênico-sanitária da água de consumo humano em propriedades rurais por meio da contagem de indicadores microbiológicos de potabilidade. MÉTODOS: Foram colhidas 180 amostras de água utilizada para consumo humano das fontes, reservatórios e ponto de consumo em 30 propriedades rurais, situadas na região Nordeste do Estado de São Paulo. Determinou-se o número mais provável de coliformes totais, Escherichia coli e o número de microrganismos mesófilos. Foi verificada a presença de medidas de proteção das fontes de abastecimento. RESULTADOS: Os resultados evidenciaram que 90% das amostras de água das fontes, 90% dos reservatórios e 96,7% de água de consumo humano, colhidas no período de chuvas, e 83,3%, 96,7% e 90%, daquelas colhidas respectivamente nos mesmos locais, durante a estiagem, estavam fora dos padrões microbiológicos de potabilidade para água de consumo humano. CONCLUSÕES: A água utilizada nas propriedades rurais foi considerada um importante fator de risco à saúde dos seres humanos que a utilizam. A adoção de medidas preventivas, visando à preservação das fontes de água, e o tratamento das águas já comprometidas são as ferramentas necessárias para diminuir consideravelmente o risco de ocorrência de enfermidades de veiculação hídrica.

Evolução de títulos sorológicos, nas provas de soroaglutinação em placa, antígeno acidificado tamponado e fixação de complemento, em bezerras Nelore vacinadas aos 18 meses de idade com Brucella abortus amostra B 19

Com o objetivo de estudar a persistência da resposta sorológica, através das provas de soroaglutinação em placa, rosa Bengala e fixação de complemento, 108 bezerras, com idade ao redor de 18 meses, foram vacinadas com uma dose padrão da vacina preparada com Brucella abortus amostra B 19. Foram obtidas amostras de soro sangüíneo antes da vacinação e após 45 dias, 6, 12 e 18 meses. Antes da vacinação, todas as bezerras apresentaram resultados negativos nos três testes. Após 45 dias, todas apresentaram título a partir de 1/100 na prova de soroaglutinação em placa e todas apresentaram resultado positivo no teste rosa Bengala, ao passo que dois animais apresentaram título 1/2 e os demais apresentaram título a partir de 1/4 na reação de fixação de complemento. Após um ano de vacinação, a grande maioria das bezerras já não apresentava título sorológico significativo. Considerando o risco a que estão sujeitos animais que vivem em áreas endêmicas e em propriedades onde a doença ocorre, e considerando a acentuada redução de título sorológico observada na grande maioria dos animais, pode-se concluir que, no caso bezerras de raças zebuínas em áreas endêmicas e que não tenham sido vacinadas na idade regulamentar, é mais vantajoso vaciná-las com a amostra B 19 aos 18 meses de idade do que deixá-las expostas a um elevado risco de infecção por Brucella.

Estabilidade do antígeno de célula total de Brucella abortus para uso no diagnóstico sorológico da brucelose bovina pela reação de fixação de complemento

A reação de fixação de complemento é um dos testes usados no diagnóstico confirmatório da brucelose bovina, e para sua realização emprega-se o mesmo antígeno usado na prova de soroaglutinação lenta, porém não foi possível encontrar na literatura estudos sobre a estabilidade desse antígeno para uso na prova de fixação de complemento, de modo a estabelecer um prazo de validade para o mesmo. Por isso, esta investigação teve por objetivo avaliar a estabilidade do antígeno de célula total de Brucella conservado sob refrigeração, para uso na reação de fixação de complemento. Analisaram-se 14 partidas de antígeno, preparado com Brucella abortus amostra 1119/3 e padronizado para uso na prova de soroaglutinação lenta, com tempo de fabricação variando de 9 meses a 23 anos e 11 meses. Testaram-se 167 soros bovinos com títulos variáveis de anticorpos contra Brucella, adotando-se a técnica com incubação a 37ºC nas duas fases da reação e 5 unidades hemolíticas 50% de complemento. Considerou-se como positivo o soro com pelo menos 25% de fixação de complemento na diluição 1:4. Compararam-se os resultados obtidos com as 13 partidas de antígeno com aqueles obtidos com a partida com 9 meses de fabricação, usando o teste de chi2 de McNemar e o coeficiente kappa. A grande maioria dos soros apresentou resultados muito próximos quando testados com as diversas partidas de antígeno, e não se observou relação entre tempo de fabricação do antígeno e diferenças nos resultados obtidos.