Câncer do colo uterino: rastreamento, marcador biológico e polimorfismo genético

Existem fortes evidências de que os programas de rastreamento baseados em citologia resultaram em diminuição significativa da incidência e mortalidade por câncer do colo do útero, no entanto, um excesso substancial de tratamento de lesões intraepiteliais de baixo grau que dificilmente progrediriam para carcinoma cervical resulta da baixa especificidade do tradicional rastreio citológico. A detecção precoce das lesões através do rastreamento citológico e a avaliação do grau histológico em espécimes cervicais são fundamentais, entretanto não permitem identificar quais pacientes terão maior probabilidade de progressão para lesões de alto grau e carcinoma invasivo. A busca de potenciais marcadores de prognóstico; objetivando o entendimento da progressão das lesões intraepiteliais é de suma importância. Acredita-se que fatores imunoregulatórios, imunogenéticos e proteínas do ciclo celular estejam intimamente envolvidos no processo de carcinogênese. Considerando o exposto, a proposta do projeto foi inicialmente avaliar a eficácia da citologia oncológica no rastreamento do câncer cervical, foi investigado ainda o polimorfismo do gene do fator de transcrição FOXP3 e a expressão da proteína do ciclo celular P63 (P63) associados respectivamente a diagnóstico e prognóstico das lesões cervicais. Em um primeiro momento foi realizado estudo transversal que envolveu 3194 mulheres. As participantes foram submetidas à citologia e biópsia de colo dirigida por colposcopia e os resultados foram comparados para verificar-se a acurácia do teste de Papanicolaou na detecção de lesões intraepiteliais e câncer cervical. Posteriormente, realizou-se estudo comparativo do tipo observacional estratificado em três grupos: Grupo 1: 16 casos com diagnóstico histopatológico de metaplasia/cervicite, considerados normais, Grupo 2: 11 casos com lesão de baixo grau (LSIL) e Grupo 3: 15 casos com lesão de alto grau (HSIL) ou carcinoma epidermoide de colo. Um total de 42 participantes respondeu a um questionário epidemiológico padronizado sobre as características demográficas, hábitos pregressos, história reprodutiva e de comportamento sexual. Após exame colposcópico, foram coletados fragmentos de espécimes cervicais para a pesquisa da expressão proteica da P63 por imunohistoquímica. Amostras de sangue periférico foram coletadas para extração do DNA e detecção do polimorfismo do gene FOXP3. No primeiro estudo em que se avaliou a acurácia do teste de Papanicolaou, encontrou-se sensibilidade de 0,83, valor preditivo positivo (VPP) de 0,77 e especificidade de 0,23 no rastreamento das lesões cervicais e câncer de colo. viii Melhores resultados foram observados quando se avaliou a acurácia diagnóstica para lesões de alto grau e carcinoma com VPP de 0,99 e especificidade de 0,84.No estudo subsequente onde se comparou a expressão da proteína P63 observou-se maior número de núcleos marcados no grupo com lesões intraepiteliais de alto grau e câncer quando comparado ao grupo com biópsias negativas (p=0,0004). No último estudo pesquisou-se a associação do polimorfismo do gene FOXP3 com lesões intraepiteliais cervicais sendo evidenciada maior prevalência do genótipo heterozigoto, CT, no grupo com lesões de colo na histopatologia (p=0,027). Mulheres com lesões intraepiteliais de baixo ou alto grau e câncer de colo de útero apresentam maior expressão da proteína P63 e maior prevalência de genótipo heterozigoto do gene FOXP3 em comparação com as sem lesões cervicais. A associação da pesquisa da expressão da proteína e do polimorfismo do gene pode tornar os exames utilizados atualmente para a avaliação diagnóstica e prognóstica das lesões de colo uterino mais efetivos em detectar quais as mulheres com maior risco para progressão para câncer

Bioprospecção de extratos metanólicos de macroalgas marinhas do litoral do Rio Grande do Norte: avaliação das atividades anticoagulante, antioxidante e antiproliferativa

Seaweeds are organisms known to exhibit a variety of biomolecules with pharmacological properties. The coast of Rio Grande do Norte has over 100 species of seaweeds, most of them not yet explored for their pharmacological potential. Sugars and phenolic compounds are the most studied of these being assigned a range of biological properties, such as anticoagulant , antiinflammatory, antitumor and antioxidant activities. In this work, we obtained methanolic extracts from thirteen seaweeds of the coast of Rio Grande do Norte (Dictyota cervicornis; Dictiopterys delicatula; Dictyota menstruallis; D. mertensis; Sargassum filipendula; Spatoglossum schröederi; Acanthophora specifera; Botryocladia occidentalis; Caulerpa cupresoides; C. racemosa; C. prolifera; C. sertularioides e Codium isthmocladum). They were evaluated as anticoagulant and antioxidant drugs, as well as antiproliferative drugs against the tumor cell line HeLa. None of the methanolic extracts showed anticoagulant activity, but when they were evaluated as antioxidant drugs all of extracts showed antioxidant activity in all tests performed (total antioxidant capacity, sequestration of superoxide and hydroxyl radicals, ferric chelation and reductase activity), especially the algae D. mentrualis, D. cilliolata and C. prolifera, who had the greatest potential to donate electrons.In addition, the ability of iron ions chelation appears as the main antioxidant mechanism of the methanolic extracts of these seaweeds mainly for the extract of the C. racemosa seaweed, which reached almost 100% activity. In the MTT assay, all extracts showed inhibitory activity at different levels againts HeLa cells. Moreover, D. cilliolata (MEDC) and D. menstrualis (MEDM) extracts showed specific activity to this cell line, not inhibiting the viability of 3T3 normal cell line, so they were chosen for detailing the antiproliferative mechanism of action. Using flow cytometry, fluorescence microscopy and in vitro assays we demonstrated that MEDC and MEDM induced apoptosis in HeLa cells by activation of caspases 3 and 9 and yet, MEDC induces cell cycle arrest in S phase. Together, these results showed that the methanolic extracts of brown seaweed D. menstrualis and D. cilliolata may contain agents with potential use in combatting cells from human uterine adenocarcinoma. This study also points to the need for more in-depth research on phytochemical and biological context to enable the purification of biologically active products of these extracts

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

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.

Avaliação in vitro da eficácia de produtos homeopáticos contendo momordica charantia através de bioensaios

Homeopathic medicines have been used for over two hundred years without the examination of their effects on in vivo and in vitro assays, due to the peculiarity of homeopathic preparations, the high dilution, which creates a challenge for the use of usual analytical techniques of quality control of medicine.Although there is scarcity of literature and variety of experiments, recently there have been some studies with few in vitro assays which have shown positive responses when evaluating the mechanism of action of homeopathic medicines which are able to act on a specific system.The present study aims to evaluate the efficacy of homeopathic products containing Momordica charantia through bioassays.Homeopathic products were tested by the MTT to assess cytotoxicity in RAW 264.7 (macrophage-like cells) and in tumor cells HeLa (human cervical adenocarcinoma cells), CHO K1 (Chinese hamster ovary cells), PANC-1 (human pancreas cancer cells) and PC-3 (human prostate cancer cells), dosage of inflammatory mediators NO, TNF-α and IL-6 released by RAW 264.7 cells, analysis of the death process and cell cycle changes of PC-3 by flow cytometry. The data demonstrate that homeopathic products of Momordica charantia did not show cytotoxicity to RAW 264.7, increased the production of inflammatory mediators by RAW 264.7 synergistically with LPS, showed cytotoxicity to PC-3 with change in its cell cycle inhibiting its proliferation, being the 30CH the most potent sample. Correlation studies were conducted in order to evaluate the possible in vitro applicable models to the quality control of homeopathic products with Momordica charantia. The data showed that the best applicable models in assessing the quality are the MTT to assess cytotoxicity in RAW 264.7 and PC-3 in 24 hours for Momordica charantia fruit products and dosage of NO production by RAW 264.7 with and without LPS

Avaliação da atenção humanizada ao abortamento em maternidade-escola, em Natal, Rio Grande do Norte

Unplanned pregnancy is experienced by millions of women worldwide. Such fact increases the risk of abortion-related morbimortality, which represents a serious public health problem. This study aims to evaluate the advances and challenges of the implementation of Humanized Abortion Care at the Maternity-School in Natal, state of Rio Grande do Norte. The research was evaluative, was preceded by an Evaluative Study, and resulted in a Case Study. The intentional sample totaled 102 subjects (60 users, 39 professionals and 3 managers). The collection techniques included documental analysis, semi-structured interview and observation with a field diary. The documental analysis was descriptive, while the Content Analysis by Bardin was used for semi-structured interviews and field diary. The Evaluative Study observed that Humanized Abortion Care is an evaluative program with preparation and pact of the logical model, of the matrix of indicators and evaluative questions. The Case Study showed that users were satisfied with the problem-solving capacity and access to the service; however, is also showed that they pointed out inadequacy in terms of environment, qualified hearing and reproductive planning. Professionals reported that the inefficiency of service consists of infrastructure and environment, which are considered inefficient and inadequate to humanized care, especially regarding patient accommodation, the lack of hospital beds, the reduced number of rooms in the surgical center and the lack of laboratory inside the maternity. Moreover, reproductive planning does not consist of an institutionalized practice in the service, and integrality with other services or partnership with the community is not in place. The Maternity Board emphasizes that the excessive demand of patients is one of the reasons that hinders the appropriate implementation of the technical standard. We then conclude that although satisfied regarding problem-solving capacity in terms of service and ease of access, there is room for improvement in qualified hearing systems, in the creation of a system to promote team work, implementation of ombudsman and satisfaction surveys. The right of shared choice did not prevail among users and health professionals with regard to the option of uterine evacuation procedure. Environment was the most mentioned category as that requiring more changes, seeing as a limited factor for the development of humanized and welcoming practices. Health professionals do not establish a periodic routine of planning practices, and such practices are not aligned with the Technical Standard. Incorporation of guidelines and availability of a plurality of methods and possibilities of choices for family planning are required. There is no institutionalization of reference and counter-reference, or partnerships with the community, which makes integrality of care not viable. The Standard needs to be included in the action plans of managers as one of the priorities in the construction of care strategies for women's health, in order to enable, allied to other initiatives, the real integration among safe conduct service, primary care network and social organizations. As a result, respect for human rights and adequate humanized care, as a way of attention and prevention of abortion, can be secured.