Fatores cl?nicos, comportamentais, nutricionais, polimorfismo C677T do gene da enzima metilenotetraidrofolato redutase (MTHFR) e risco de c?ncer de mama.

Programa de P?s-Gradua??o em Ci?ncias Biol?gicas. N?cleo de Pesquisas em Ci?ncias Biol?gicas, Pr?-Reitoria de Pesquisa e P?s Gradua??o, Universidade Federal de Ouro Preto.O C?ncer de Mama (CM) ? uma doen?a com taxas de incid?ncia crescentes em diversas partes do mundo e tamb?m no Brasil, a qual ? causada pela intera??o de diversos fatores de risco. Foi realizado estudo caso controle, baseado em hospital, mascarado, com 654 mulheres (com CM, controles e com doen?a ben?gna da mama) para avaliar as intera??es entre diversos fatores de risco e a ocorr?ncia de CM. A MTHFR, envolvida no metabolismo do ?cido f?lico, catalisa a redu??o da 5,10-MTHF a 5- MTHF que ? necess?rio para a convers?o de homociste?na em metionina pela metionina sintase. Uma redu??o da atividade da MTHFR produz uma enzima termol?bil, resultando em um aumento da homociste?na plasm?tica e do estado de hipometila??o gen?mica. Estudos epidemiol?gicos sugerem que o polimorfismo do gene da Metilenotetrahidrofolato redutase (MTHFR), C677T, pode modificar o risco para CM. Foi realizada antropometria e aplicados question?rios para avalia??o de caracter?sticas s?cio-demogr?ficas, cl?nicas, antropom?tricas, reprodutivas, diet?ticas, comportamentais e da freq??ncia de consumo alimentar (para avaliar a ingest?o de ?cido f?lico, vitaminas B6 e B12). Foi coletado sangue perif?rico do paciente para extra??o do DNA. A genotipagem do polimorfismo C677T foi realizada atrav?s da t?cnica PCR-RFLP. Atrav?s do modelo log?stico criado para explicar a ocorr?ncia do CM de mama na popula??o estudada, foi observado que o sedentarismo, a baixa renda, a n?o amamenta??o e a menopausa, ajustados pela idade, uso de CTO, % GC, escolaridade e idade da primeira MMG afetaram de forma estatisticamente significativa o risco para a ocorr?ncia do CM. Mulheres na pr?-menopausa que n?o amamentaram seus filhos apresentaram risco de 2,15 vezes de desenvolver CM em rela??o ?s mulheres na p?s menopausa. Mulheres etilistas na p?s-menopausa apresentaram maior risco de desenvolver CM em rela??o ?s mulheres n?o etilistas na p?s menopausa. O consumo de ?cido f?lico encontrou-se acima da recomenda??o pr?-estabelecida pela RDA, por?m foi observado um consumo menor, n?o significante, no grupo de casos quando comparado com o grupo controle. Foi observado que o consumo de vitamina B6 se encontrava abaixo da recomenda??o no grupo de casos e, portanto este baixo consumo pode interferir no metabolismo do folato e da enzima da MTHFR. O polimorfismo C677T de MTHFR nem isoladamente nem em associa??o com fatores comportamentais e diet?ticos neste trabalho n?o foi suficiente para aumentar o risco de CM, diferente do sugerido por resultados de outros trabalhos.The breast cancer (BC) is a disease, with increasing incidence rate worldwide and also in Brazil, which is caused by the interaction of different risk factors. A masked casecontrol study was conducted with 654 women (with breast cancer, controls and with benign breast disease) to assess the interaction between several risk factors and the breast cancer occurrence. The methylenetetrahydrofolate reductase (MTHFR), involved in the folic acid metabolism catalyzes the 5,10-MTHF to 5- MTHF reduction, necessary to convert homocystein into methionine by methionine synthase. A reduction in the activity of MTHFR produces a thermo labile enzyme increasing plasma homocysteine and leading to genomic hypomethylation. Epidemiologic studies suggest that the MTHFR C677T gene polymorphism may modify the risk of BC. Anthropometric measures were collected and surveys for evaluation of social-demographic, clinical, behavioral features and food frequency ingestion (folic acid, vitamins B6 and B12) were conducted. Peripheral blood was collected for DNA extraction. The C677T polymorphism genotyping was conducted using a PCR-RFLP protocol. The logistic model created to explain the occurrence of BC has shown that sedentary behavior, low incomings, no breast-feeding and menopause, adjusted by age, oral contraceptive use, %body fat, education level and date of the first mammography has affected the risk for BC occurrence with statistic significance. Women in pre-menopause that did not breast-feed their children has a 2,15 times higher risk of BC compared with women in post-menopause. Alcoholic women in post-menopause presented a higher risk for BC compared to non-alcoholic women in post-menopause. The folic acid ingestion has shown to be higher than the established RDA recommendation, although the BC group presented a lower ingestion compared with the control group. The ingestion of vitamin B6 was lower than the recommendation in the case group, and this low ingestion can interfere in the folate metabolism. In this work, the C677T polymorphism, neither isolated, nor in association with behavioral and dietary factors, has not affected the risk of BC, opposed to results suggested in other works

Importância prognóstica dos níveis transcricionais do gene TNRC6A em pacientes com Câncer de Mama / Prognostic relevance of transcriptional levels of TNRC6A gene in patients with Breast Cancer

O câncer de mama (CAM) é o câncer mais comumente diagnosticado entre as mulheres no mundo ocidental, com um número crescente de óbitos. A busca de marcadores tumorais na avaliação do prognóstico dessa doença tem se tornado crucial para a escolha terapêutica, o que impacta diretamente a sobrevida das pacientes. A proteína TNRC6A tem sido descrita associada às vias de miRNA, regulando toda a maquinaria celular de células normais e tumorais. O presente estudo tem como objetivo avaliar a expressão dos transcritos do gene TNRC6A e sua correlação com a ocorrência e progressão do CAM. Foram coletadas 43 amostras de sangue periférico de pacientes com CAM e 44 com Doença Benigna da Mama (DBM). Em cada amostra foi realizada a transcrição reversa seguida de ensaios de qPCR. Com a análise dos níveis de mRNA verificou-se que não houve uma diferença significativa entre o grupo CAM e DBM. Além disso, os dados de idade, receptores hormonais e proliferação celular também não mostraram associação com o marcador. Contudo, menores níveis foram identificados em pacientes com comprometimento dos gânglios linfáticos axilares, evidenciando a importância do gene TNRC6A na progressão de tumores mamários.

Viability and gene transcription of Campylobacter jejuni in infant milk formulas

This study simulated the contamination of two varieties of infant milk formulas (homemade and commercial) with 103 and 104 CFU/mL of Campylobacter jejuni, that were kept under refrigeration (4-7ºC) for up to 48 hours. The aim of this study was to verify the maintenance of the viability and ability of Campylobacter jejuni to produce transcripts of virulence and resistance to stress conditions during periods of 0 (after preparation), 24 and 48 hours. C. jejuni remained viable during all analyzed stages and the presence of coliforms was not detected. In general, the counts reduced 1 log cycle after 48 hours for all samples, except the 104 CFU/mL inoculum of commercial formula, which reduced 2 log cycles, indicating greater injury of C. jejuni in this food matrix. C. jejuni showed to be more adapted to homemade matrix, due to high transcription of the gene related to cell invasion, ciaB, and more susceptible in the commercial matrix, due to the high transcription  of genes related to conditions of stress tolerance (dnaJ, p19, sodB). The low infective dose of C. jejuni coupled with greater vulnerability of children less than five years indicate the need for care in the preparation and maintenance of infant formulas, to prevent the use of contaminated raw material and cross-contamination, especially in homemade formulations

Prospective Analysis of Food Consumption and Nutritional Status and the Impact on the Dietary Inflammatory Index in Women With Breast Cancer During Chemotherapy

Considering the implications of adverse effects of chemotherapy (CT) and the potential impact of diet on patients’ recovery, this study aimed to prospectively evaluate the association between the consumption of food groups, patients’ Dietary Inflammatory Index (DII®) scores, and their nutritional status. Anthropometric and dietary assessments of 55 women with breast cancer (BC) were performed at three time points. T0 is the time point after the first CT cycle, T1 is the time point after the intermediate CT cycle, and T2 is the time point after the last CT cycle. We identified a significant increase in weight, body mass index, and waist circumference during CT. Consumption of poultry and eggs was higher in T1 when compared to T2, while consumption of total fruit and total vegetables was higher at T0 compared to T1 and T2. The diet became more pro-inflammatory over the course of treatment (X2(2) = 61.127), and was related to higher abdominal adiposity. Total fruit (T0: R2 = 0.208, T1: R2 = 0.095, T2: R2 = 0.120) and total vegetable consumption (T0: R2 = 0.284, T1: R2 = 0.365, T2: R2 = 0.580) predicted DII® change at the three-time points. Meanwhile, consumption of total grains was significantly associated only with T1 (R2 = 0.084) and T2 (R2 = 0.118), and consumption of simple sugars was significantly associated only with T0 (R2 = 0.137) and T1 (R2 = 0.126). Changes in food consumption led to an increase in the inflammatory profile of the diet, suggesting the necessity to improve the guidelines during and after CT. These results reinforce the need to promote healthier eating practices in concert with maintaining a healthy nutritional status in women with BC treated with C

Consumption of minas frescal cheese may be a source of human infection by Campylobacter jejuni

Campylobacter spp. is an emerging pathogen that causes gastroenteritis in humans and the consumption of dairy food can characterize sources of infection. We aimed to verify the viability and a presence of transcripts associated with characteristics of virulence and adaptation of C. jejuni isolated from Minas Frescal cheeses, produced with contaminated milk and stored under refrigeration for up to ten days. The samples were analyzed for bioindicators, Campylobacter spp., pH, acidity, moisture and sodium chloride. Campylobacter spp. recovered were evaluated for the production of transcripts of: ciaB, dnaJ, p19 and sodB. The results were correlated with the viability of C. jejuni and changes in their transcriptome. Storage at low temperatures reduced C. jejuni from the first to the fourth day. The variations in humidity, pH and acidity influenced the decreasing of C. jejuni. There was a reduction in transcripts' production of the four genes, more pronounced on the fourth day, indicating the inability of the microorganism to perform its metabolic activities, due to the conditions of injury. Despite the presence of mechanisms of virulence and adaptation, C. jejuni could not remain viable four days after production. However, consumption of fresh cheese contaminated with Campylobacter jejuni can be a source of infection when consumed up to four days after production

Pept?deos ligantes de c?lulas tumorais e de imunoglobulinas g espec?ficos do c?ncer de mama.

At? o momento, n?o existem biomoleculares que sejam efetivamente utilizados no diagn?stico do c?ncer de mama. Uma alternativa para auxiliar a busca por marcadores moleculares ? baseada na sele??o de ligantes de alta afinidade por meio de Phage Display (PD). O principal objetivo desta tese foi o desenvolvimento de novos biomarcadores envolvidos no c?ncer de mama selecionados contra uma linhagem celular tumoral altamente agressiva e contra IgG purificada espec?fica de tecido mam?rio de pacientes. Ap?s a sele??o PD em biblioteca aleat?ria de pept?deos, todos os clones imunorreativos foram caracterizados por sequenciamento, deduzidos in vitro e submetidos a an?lises in silico. Valida??es posteriores foram conduzidas com ELISA e an?lise histopatol?gica. Apesar de terem sido relatadas diversas varia??es da metodologia, a maior parte dos processos de sele??o usou ligantes obtidos depois de tr?s ciclos de sele??o. Os marcadores relatados na literatura foram pouco explorados devido ? complexidade dos processos de valida??o. Este trabalho demonstrou que as sele??es do primeiro e segundo ciclos s?o vi?veis e podem ser utilizadas para aumentar o n?mero de potenciais marcadores a serem testados. Os resultados com linhagem de c?lulas tumorais identificaram tr?s potenciais marcadores no terceiro ciclo de sele??o, dos quais foi selecionado aquele com melhor rela??o de absorb?ncia tumor/controle no ELISA para s?ntese qu?mica. O pept?deo sint?tico foi associado com uma marca??o menos intensa em tumores triplo negativos (Receptor de Estr?geno, Receptor de Progesterona e ErbB2). Outros pept?deos do segundo ciclo tamb?m demonstraram especificidade a tecidos tumorais. Para a estrat?gia de mapeamento de ant?genos contra IgG purificada espec?fica de tecido mam?rio, um pept?deo (F4) pode reconhecer tanto tecidos tumorais de mama quanto IgG circulante. Ambos clone F4 e pept?deo sint?tico SF4 alcan?aram diagn?stico de alta acur?cia, mas surpreendentemente, o clone F4 apresentou a melhor sensibilidade e especificidade (77,8% e 85,7%, respectivamente), sugerindo que pode ser utilizado no diagn?stico para triagem precoce de c?ncer de mama, anterior ?s an?lises por imagem e patol?gicas. A identifica??o e caracteriza??o do biomarcador putativo est? em investiga??o por meio de sele??o combinatorial de anticorpos scFv contra o fago F4 e o pept?deo SF4, os quais ser?o utilizados para captura imunol?gica do ant?geno verdadeiro e para an?lise por espectrometria de massas. Um sensor eletroqu?mico de baixo custo utilizando o pept?deo SF4 tamb?m foi desenvolvido.Currently there are no biomarkers that have effectively been used in breast cancer (BC) diagnosis, which has been accomplished mainly based on mammography and pathological analysis of biopsy specimens. An alternative technology for the discovery of markers is through selection of high affinity ligands by Phage Display (PD). The main objective of this investigation was the development of new biomarkers involved in breast cancer that were selected against a highly aggressive tumor cell lineage and against purified tissue-specific IgG from BC patients. After PD selections of random peptide libraries, all immunoreactive ligands were further characterized by sequencing, in vitro translated and submitted to bioinformatic analyses. Further validations were performed by ELISA and immunohistochemistry. Although many variations of the methodology have been reported, most of the selection processes have used ligands obtained after three or more cycles of selection, but successful markers have been marginally explored due to complex validation processes. We have demonstrated that selections in the first and second cycles are feasible and can be used to increase the number of potential markers to be tested. Furthermore, results from the tumor cell lineage also identified three potential peptide markers in the third cycle that presented the best relation was chosen for peptide synthesis. The synthetic peptide was associated to less intense labeling of triple negative tumors (Estrogen Receptor, Progesterone receptor, and ErbB2). Other peptides from the second cycle also seemed very specific to tumor tissues. For the antigen mapping strategy against purified tissue-specific IgG from patients? tumor biopsies, one peptide (F4) could recognize both breast tumor tissues as well as circulating IgG from patients. Both F4 clone and its synthetic peptide reached a very high accurate diagnosis, but surprisingly, the selected F4 clone presented the highest sensitivity and specificity, (77.8% and 85.7%, respectively), suggesting that it can be used as a diagnostic reagent for early BC screening prior to imaging and pathological analyses. Identification and characterization of the putative biomarker is under investigation through Fab combinatorial antibody selection against both F4 clone and peptide, which will be used for immunological capture of the true antigen and for mass spectrometry analysis. A low cost electrochemical sensor using the SF4 peptide was also developed

Does sleep influence weight gain during pregnancy? A prospective study

Objective: The focus of this study was to evaluate the associations between subjective sleep quality and duration and weight gain during pregnancy. Methods: A prospective and longitudinal study was conducted with 63 pregnant women. Pregnant women were evaluated at the first, second and third trimester for subjective sleep quality (Pittsburgh Sleep Quality Index [PSQI]) and anthropometric variables for body mass index [BMI] calculation. The sleep quality was grouped per cluster, identifying those individuals who maintained, improved or worsened their sleep quality, based on the PSQI classifications. Generalized estimating equations (GEE) were used to examine the association between sleep and BMI over the pregnancy period. Results: An effect of the interaction between time of pregnancy and clusters of sleep quality was observed on the BMI (p<0.05), which indicates that pregnant women who improved subjective sleep quality during pregnancy gained more weight from the second to third trimester, while those that worsened the subjective sleep quality gained more weight during the first to second trimester. Sleep duration was not associated with weight gain. However, pregnant women who maintained the same BMI category over the pregnancy period increased their sleep duration from the first to third trimester, while those that increased the BMI category slept the same amount of time during this period (median=1.0 [0.0–2.0] and median=0.0 [-2.0–1.0], respectively, p=0.039). Conclusions: The authors concluded that a worse subjective sleep quality seems to lead to an inadequate weight gain distribution during the period of pregnancy

Relationship between food perceptions and health-related quality of life in a prospective study with breast cancer patients undergoing chemotherapy

OBJECTIVE: To correlate the perceptions related to dietary intake with the domains and subscales of health-related quality of life (HRQL) in women with breast neoplasms receiving chemotherapy. METHODS: In this prospective study, 55 women with breast cancer were followed up during chemotherapy at three different times (T0, T1, T2). Before chemotherapy, perceptions related to food consumption were evaluated. HRQL was analyzed with the EORTC QLQ-C30 and Br23 instruments 21 days after each investigated cycle. The differences (T2-T0) in the subscales and HRQL domains were correlated with the differences (T2-T0) in the appetite scores. Spearman’s correlation was used to verify a possible correlation between differences in functional and overall HRQL domains (T2-T0) and differences in appetite scores for certain foods and between the differences in some subscales of EORTC QLQ-C30 and Br23 (T2-T0) and differences in appetite scores for certain food groups (T2-T0). RESULTS: Correlations between pain and appetite for bitter taste and between an increased appetite for juices and pain intensification or fatigue were identified, and pain was correlated with an appetite for starchy foods. An appetite for vegetables, legumes and meat/eggs was correlated with physical function. The only significant correlation with social functions occurred between the appetite for sweet foods and these functions. We found a correlation between overall health, emotional function, social function and physical function and the appetite for juices. CONCLUSION: Chemotherapy alters the individual’s relationship with food and, consequently, the individual’s HRQL

Temporal influence of endocrine therapy with tamoxifen and chemotherapy on nutritional risk and obesity in breast cancer patients

Abstract Background The effect of endocrine therapy with tamoxifen (TMX) on weight gain has been reported in the literature, but the outcomes are still controversial. Moreover, previous treatment options, such as chemotherapy (CT), also include body changes. The focus of this study was to verify the temporal influence of endocrine therapy with TMX on nutritional risk and obesity and its association with CT in breast cancer patients. Methods In this cross-sectional study, 84 breast cancer surviving women were evaluated during endocrine therapy with TMX. Anthropometric, biochemical and body composition parameters were measured. A generalized estimating equation (GEE) was used to examine the association between CT and groups of women using TMX categorized by the duration of the treatment (group 1, women using TMX for the first 3 years; group 2, women using TMX between 3 and 4 years and group 3, women using TMX for more than 4 years). Results The interaction of CT with duration of TMX use showed a significant effect on Body Mass Index (BMI), waist circumference (WC) and body fat percentage (BFP) (GEE p-value = 0.002, 0.000, 0.000, respectively). Women from group 1 who underwent CT presented higher values of body variables compared to those women from group 2 who also underwent CT (BMI = 29.14 ± 0.93, 26.76 ± 0.85 kg/m2; WC = 94.45 ± 1.96, 91.07 ± 2.44 cm; BFP = 36.36 ± 1.50, 33.43 ± 1.66%, respectively). On the other hand, women from group 1 who did not undergo CT presented lower values of body variables compared to those women from group 2 who also did not undergo CT (BMI = 25.29 ± 0.46, 28.40 ± 0.95 kg/m2; WC = 85.84 ± 0.90, 97.75 ± 0.88 cm; BFP = 30.32 ± 0.43; 42.95 ± 1.03%, respectively). Conclusions Women on endocrine therapy with TMX are mostly overweighed and obese, most evidently in women who received CT, and who were at the beginning of treatment. Women that did not undergo CT, despite presenting lower values of body variables in the first 3 years, still deserve special attention because significantly higher values were observed in women between 3 and 4 years of therapy