Claudia Giuliano Bica

MnSOD Gene Polymorphism Association with Steroid-Dependent Cancer

Oxidative stress enhances carcinogenesis due to DNA damage. Manganese superoxide dismutase (MnSOD) Val16Ala polymorphism has been recently associated with breast and prostate cancer. The role of oxidative stress in male breast cancer is poorly investigated due to the low prevalence of this neoplasia. We studied the relationship between prostate cancer (PC), male (MBC) and female breast cancer (FBC) and this polymorphism in a case–control study. Human genetic polymorphism Val16Ala of MnSOD was obtained from blood and paraffin-embedded tumor samples. The polymorphism was determined in 11 cases of MBC, 51 cases of PC, 89 cases of FBC and 372 age-adjusted healthy controls by polymerase chain reaction–restriction fragment length polymorphism techniques using restriction enzyme Hae III. Chi-square or Fisher test were used to compare the MnSOD frequency distribution. The observed genotypic frequencies of all samples were AA = 9.6% (n = 50), VV = 25.4% (n = 133) and AV = 64% (n = 340), all at Hardy–Weinberg equilibrium. Breast and prostate cancer risk was elevated in male and female patients with the Ala/Ala genotype compared to controls (p = 0.006, odds ratio = 2.5, 95% confidence interval 1.393–4.541). Even though the frequency of the Ala allele was low (9.6%) in the studied population, these data support the hypothesis that MnSOD and oxidative stress play a significant role in breast cancer risk both in males and females and also brings new information on the role of this polymorphism in prostate cancer. This is the first study which provides some evidence that genetic polymorphism in the MnSOD gene may be associated with an increased risk of male breast cancer. Studies with a larger sample size are needed to confirm the findings.

Polymorphism (ALA16VAL) correlates with regional lymph node status in breast cancer

We studied the possible association between Ala16Val manganese-dependent superoxide dismutase (MnSOD) gene genotypes and breast cancer lymph node status because previous investigations suggested an association between the AA genotype and breast cancer. We included 281 women (188 controls and 93 cases of invasive breast cancer with axillary lymph node metastasis (LN+) and without lymph node metastasis (LN-). DNA was extracted from paraffin-embedded tumor tissue or peripheral blood leukocytes, and MnSOD polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism techniques. In addition, the immunohistochemical profile (p53, Ki-67 and estrogen/progesterone receptors) was also compared between invasive breast cancer groups and different MnSOD genotypes. The frequency of the VV genotype was higher in the LN+ group than in the control and LN- groups (chi(2)=5.081, P=0.02). Subjects with LN+ breast cancer (LN+ group) showed a higher incidence of VV genotype carriers associated with positive Ki-67 marker. Subjects with LN+ breast cancer (LN+ group) showed a higher incidence of VV genotype carriers associated with negative p53 marker. Despite the fact that the AA genotype is well established as being associated with an increased risk of breast cancer, the VV genotype may be associated with a higher metastatic potential, suggesting that MnSOD imbalance is the condition associated with carcinogenesis.

Fatores clínicos e anatomopatológicos que influenciam a sobrevida de pacientes com câncer de mama e derrame pleural neoplásico

OBJECTIVE: The objective of this study was to identify the clinical and pathological factors that can influence the prognosis of breast cancer patients with clinical symptoms of malignant pleural effusion. METHODS: This was a clinical cohort study, in which we analyzed the medical charts of patients diagnosed with malignant pleural effusion between 2006 and 2010. By examining the charts, we identified the female patients with a history of breast cancer. For those patients, we collected pathology data related to the primary tumor and cytopathology data related to the pleural metastasis. RESULTS: We evaluated 145 patients, 87 (60%) of whom had tested positive for malignant cells in the pleural fluid. Ductal histology was observed in 119 (82%). The triple-negative breast cancer phenotype was seen in 25 cases (17%). Those patients had the worst prognosis (with a sharp decline in the survival curve), and 20 of the 25 (80%) died during the follow-up period (through June of 2011). The mean survival after the identification of malignant pleural effusion was 6 months. CONCLUSIONS: In patients with triple-negative breast cancer who test positive for malignant cells in the pleural fluid, the prognosis is poor and survival is reduced.

Association of manganese superoxide dismutase gene polymorphism (Ala-9Val) and breast cancer in males and females

INTRODUCAO: Uma das diversas rotas metabolicas envolvidas no processo de carcinogenese da mama e o polimorfismo Ala-9Val do gene da superoxido dismutase dependente de manganes, cuja associacao com o aumento do risco de câncer de mama em mulheres ja e bem estabelecida na literatura. Contudo, nao existem estudos envolvendo esse polimorfismo no carcinoma de mama em homens, principalmente devido a baixa prevalencia dessa neoplasia. O objetivo deste estudo e analisar o polimorfismo da MnSOD em uma populacao de homens e mulheres com câncer de mama no sul do Brasil, comparando tais achados com controles saudaveis. METODOS: Foi realizado um estudo de caso-controle em cem pacientes com câncer de mama (11 homens e 89 mulheres) e 370 controles saudaveis. O DNA foi extraido do tecido tumoral emblocado em parafina. O polimorfismo da MnSOD foi determinado por tecnicas de PCR-RFLP usando a enzima de restricao Hae III. O teste do qui quadrado foi usado para comparar a distribuicao das frequencias dos polimorfismos. RESULTADOS: As frequencias genotipicas dos pacientes com câncer de mama foram AA = 15%; AV = 56%; VV = 29% e dos controles AA = 6,5%; AV = 68,1% e VV = 25,4%. Os pacientes com câncer de mama, tanto as mulheres como os homens, apresentaram frequencias significativamente mais elevadas do genotipo AA quando comparadas aos controles (p = 0,035), sugerindo associacao forte desse genotipo com o câncer de mama. O intervalo de confianca foi de 1,393-4,541 (95%) e o risco encontrado foi de 2,15 para individuos portadores do genotipo AA, quando comparados com os controles que tinham os genotipos VV e AV da MnSOD. DISCUSSAO: Esses resultados confirmam a associacao ja estabelecida do genotipo da MnSOD AA com câncer de mama em mulheres e indicam distribuicao de frequencia similar e risco aumentado na populacao masculina.

Prevalence of the hemochromatosis gene mutation in patients with nonalcoholic steatohepatitis and correlation with degree of liver fibrosis

BACKGROUND Nonalcoholic steatohepatitis is a chronic liver disease with a high prevalence in the general population and a potential to evolve into cirrhosis. It is speculated that iron overload could be associated with liver injury and unfavorable progress in affected patients. AIMS To evaluate the prevalence of mutation of the hemochromatosis gene (HFE) in patients with nonalcoholic steatohepatitis and to correlate it with histological findings in liver specimens. PATIENTS AND METHODS Twenty-nine patients with nonalcoholic steatohepatitis were evaluated. The presence of mutation in the hemochromatosis gene (C282Y and H63D) was tested in all patients and its result was evaluated in relation to hepatic inflammatory activity, presence of fibrosis, and iron overload in the liver. The control group was composed of 20 patients with normal liver function tests and 20 patients infected with the hepatitis C virus, with elevated serum levels of aminotransferases and with chronic hepatitis as shown by biopsy. RESULTS Mutation of the hemochromatosis gene (C282Y and/or H63D) was diagnosed in 16 (55.2%) patients with nonalcoholic steatohepatitis, in 12 (60%) patients with hepatitis C and in 8 (40%) patients with no liver disease. No association was found between the presence of mutation and inflammatory activity, nor with the presence of fibrosis in patients with nonalcoholic steatohepatitis. An association was found between the presence of mutation and the occurrence of iron overload in liver, but there was no association between liver iron and the occurrence of fibrosis. CONCLUSIONS The findings suggest that iron does not play a major role in the pathogenesis and progression of nonalcoholic steatohepatitis, and routine tests of the hemochromatosis gene mutation in these patients should not be recommended.

The Role of Urine Cytology for ‘Decoy Cells’ as a Screening Tool in Renal Transplant Recipients

Objective: To determine the role of urine cytology for ‘decoy cells’ as a screening tool for polyomavirus type BK (BKV) infection in renal transplant recipients. Study Design: This was a prospective cohort study of patients undergoing renal transplantation between 2006 and 2010. Results: A total of 442 patients underwent urine cytology for decoy cells, 27.8% underwent 1 examination only and 72.2% more than one. Of the 1,713 examinations reviewed, 426 (24.9%) were positive and 785 (45.8%) were negative for ‘decoy’ cells, 380 (22.2%) showed degenerated tubular cells and 122 (7.1%) were unsatisfactory for analysis. Urine cytology was found to have a specificity of 68.5%, a sensitivity of 84.6%, a positive predictive value of 21.2%, a negative predictive value of 97.8% and an overall accuracy of 69.9%. The incidence of polyomavirus nephropathy among the patients investigated was 11.8%. Of the 442 patients, 32 (7.2%) had graft loss, which was attributed to BKV nephropathy in 2 (6.2% of the 32). Conclusions: Urine cytology is an effective screening method for monitoring renal transplant patients, with high sensitivity and a high negative predictive value, and can therefore be used routinely in the follow-up of renal transplant patients.

Deteção de Mycobacterium tuberculosis em amostras clínicas por reação em cadeia da polimerase utilizando primers baseados na região intergênica plcB-plcC

OBJECTIVE To develop a system for the molecular diagnosis of tuberculosis by polymerase chain reaction (PCR), constructing primers based on the difference in gene organization of the intergenic region of phospholipase C (plcB-plcC region), which differentiates Mycobacterium tuberculosis from other mycobacteria. METHODS A PCR product of the expected size (432 bp) was obtained from M. tuberculosis and M. africanum only. A total of 33 mycobacterial isolates and 273 clinical samples from patients suspected of having tuberculosis were examined. These were used in the comparative study of the PCR technique versus culture. RESULTS For PCR versus culture, the data showed 93.8% accuracy (p < 0.0001), 93.1% sensitivity (CI: 88.7-96.0), and 96.4% specificity (CI: 96.1-99.4). The Kappa value (0.82) shows that there was a near-perfect concordance between the two tests. CONCLUSION The use of the plcB-plcC region in PCR amplification was found to be an important and reliable tool for the specific diagnosis of tuberculosis in the samples analyzed.

Ethics Requirement Score: new tool for evaluating ethics in publications

Objective To analyze ethical standards considered by health-related scientific journals, and to prepare the Ethics Requirement Score, a bibliometric index to be applied to scientific healthcare journals in order to evaluate criteria for ethics in scientific publication. Methods Journals related to healthcare selected by the Journal of Citation Reports™ 2010 database were considered as experimental units. Parameters related to publication ethics were analyzed for each journal. These parameters were acquired by analyzing the author’s guidelines or instructions in each journal website. The parameters considered were approval by an Internal Review Board, Declaration of Helsinki or Resolution 196/96, recommendations on plagiarism, need for application of Informed Consent Forms with the volunteers, declaration of confidentiality of patients, record in the database for clinical trials (if applicable), conflict of interest disclosure, and funding sources statement. Each item was analyzed considering their presence or absence. Result The foreign journals had a significantly higher Impact Factor than the Brazilian journals, however, no significant results were observed in relation to the Ethics Requirement Score. There was no correlation between the Ethics Requirement Score and the Impact Factor. Conclusion Although the Impact Factor of foreigner journals was considerably higher than that of the Brazilian publications, the results showed that the Impact Factor has no correlation with the proposed score. This allows us to state that the ethical requirements for publication in biomedical journals are not related to the comprehensiveness or scope of the journal.

Impact of cell arrangement of pleural effusion in survival of patients with breast cancer.

Objective: This study was performed to evaluate the potential influence of cytological differences between pleural effusions on the survival of women with metastatic breast cancer during 30 months of follow-up. Study Design: A hospital-based cohort study was performed. Pleural fluid cytology slides from patients with breast cancer were examined. Cases were grouped according to the pattern of tumor cells (spheroid and isolated), in order to access their prognostic value. Results: The study comprised 87 patients. An isolated cell pattern was associated with higher mortality 30 months after the pleural effusion when compared to a spheroid pattern (p = 0.038). Patients with an isolated cell pattern showed higher risk of dying than patients with spheroid formations. The relative risk after adjustment of intervening variables was 5.336 (95% CI 1.054-27.020). The presence of a triple-negative immunohistochemical pattern significantly increased the risk of mortality before 30 months. Conclusion: Pleural effusion with isolated malignant cells is associated with worse prognosis after 30 months of follow-up.

Antidepressant Drugs Modulate Differentially Anti-inflammatory Lithium’s Property: An in Vitro and in Vivo Study

Li (lithium), a mood stabilizer has anti-inflammatory effect. However, in clinical practice, Li can be administered together with other antidepressants drugs, such as FLX (fluoxetine), IMI (imipramine), NOR (nortriptyline) and ESC (escitalopram). As interaction between Li and these antidepressant drugs on inflammatory modulation has not been investigated yet, we performed an in vitro protocol using a non-human macrophage cell line. Oxidative and inflammatory markers, as well as cell cycle analysis and cytokine gene expressions were compared among treatments. An IR (inflammatory ratio) was calculated based on the following oxidative-inflammatory variables: nitric oxide, superoxide anion, reactive oxygen molecules, cytokines IL-1β, IL-6, TNF-α and IL-10. The in vitro calculated IR data were validated through an in vivo analysis of 154 human subjects with similar IR. Li and control cells presented similar IR values. FLX, NOR and IMI increased slightly IR values indicating some proinflammatory effect, whereas ESC decreased IR values indicating some anti-inflammatory effect. However, cells exposed to Li + ESC triggered a proinflammatory response on macrophages. Thus, IR comparison results suggest that the Li anti-inflammatory effect is not universal and could be influenced by both basal macrophage-inflammatory state and interaction of other psychiatric drugs. These results could be useful to understand some inconsistencies observed in human studies involving Li and other psychiatric drugs.