Previous gut microbiota has an effect on postprandial insulin response after intervention with yacon syrup as a source of fructooligosaccharides: a randomized, crossover, double-blind clinical trial.

OBJECTIVES: The use of prebiotics has an effect on postprandial glucose and insulin concentrations; however, the interaction between the previous profile of the intestinal microbiota and the effect of supplementation with prebiotics is not clear. Our objective was to evaluate the effect of previous intestinal microbiota profile on the postprandial insulin response to yacon syrup, used as a source of fructooligosaccharides (FOSs), in young women. The product presents high levels of FOS. METHODS: In this double-blind, crossover, randomized clinical trial, 40 adult women were allocated to receive a breakfast containing 40 g of yacon syrup (14 g FOS, intervention A) or a breakfast containing 40 g of placebo (intervention B). On each intervention day, after 12 h of fasting, an aliquot of blood was collected for insulin analysis at 0, 15, 30, 45, 60, 90, and 120 min. The fecal sample was collected before the beginning of the interventions, and the DNA was extracted and quantified, with subsequent amplification of the 16S region, next-generation sequencing, and analysis of sequencing data. RESULTS: The glucose and insulin concentrations were reduced after ingestion of yacon syrup compared with placebo, specifically at the 30 min to insulin. After the receiver operating characteristic analysis, six volunteers who did not respond to the yacon consumption intervention were identified. The abundance of the phylum Actinobacteria (P = 0.021) and the order Bifidobacteriales (P = 0.013) were positively associated with better insulin response. Other main phyla were not associated with intervention response. CONCLUSIONS: The previous profile of the intestinal microbiota has an effect on the postprandial insulin response to FOSs, mainly in the phylum Actinobacteria and Bifidobacteriales order.

Association of angiotensin-converting enzyme I gene I/D polymorphism with endometrial but not with ovarian cancer.

Associations have been found between the angiotensin-converting enzyme insertion deletion (I/D) polymorphism (ACE I/D) and endometrial and epithelial ovarian cancer (EC and EOC, respectively). In this study, the following frequencies for each of three ACE polymorphisms, DD, ID, and II, respectively, were observed: in the EC group, 55, 24, and 21% versus the control group 39, 40, and 21% (p = 0.033*); in the EOC group 49, 36, and 15% versus the control group 49, 33, and 18% (p = 0.82). According to these allelic distributions, DD carriers are 2.0 times more likely than individuals carrying the ID or II genotypes to develop EC; therefore, the DD genotype seems to be protective against EC. In contrast, no association was observed between ACE (I/D) polymorphism with EOC. The ACE (I/D) polymorphism might play a role in the pathogenesis of EC and it should be considered when identifying genetic markers for EC.

Plasma metabolomics are associated with metabolic syndrome: A targeted approach.

OBJECTIVES: Advances in metabolomic tools have allowed us to gain a more comprehensive understanding of metabolic syndrome (MetS). The aim of this study was to evaluate the association between plasma metabolomic profiles and MetS. METHODS: For this study, adults without diabetes, chronic kidney disease, stroke, heart disease, or cancer and with full metabolomics, biochemical, and dietetic data available, representing a subsample of the Health Survey of Sao Paulo study (ISA-Capital; N = 130), were included. The joint interim statement consensus criteria were used for diagnosing MetS. Absolute quantification (µmol/L) of blood metabolites was achieved by targeted quantitative profiling of annotated metabolites by electrospray ionization tandem mass spectrometry in plasma samples. Mean differences in the compounds for MetS were evaluated by linear regression adjusted for confounding factors. RESULTS: Serine was inversely associated with MetS (ß = -15.04; P = 0.014). In glycerophospholipids with acyl-alkyl bonds, there was an inverse association with MetS, including phosphatidylcholine (PC) ae C42:5 (ß = -0.15; P = 0.040), PC ae C44:5 (ß = -0.15; P = 0.046), PC ae C40:4 (ß = -0.21; P = 0.014) and PC ae C44:4 (ß = -0.04; P = 0.032). CONCLUSION: Plasma metabolomic profiles were associated with MetS, especially the amino acid serine and some acyl-alkyl PCs.

STUDY OF LIPID BIOMARKERS OF PATIENTS WITH POLYPS AND COLORECTAL CÂNCER.

BACKGROUND: Colorectal cancer (CRC) is one of the leading causes of cancer worldwide. Early diagnostic methods using serum biomarkers are required. The study of omics, most recently lipidomics, has the purpose of analyzing lipids for a better understanding of human lipidoma. The evolution of mass spectrometry methods, such as MALDI-MS technology, has enabled the detection and identification of a wide variety of lipids with great potential to open new avenues for predictive and preventive medicine. OBJECTIVE: To determine the lipid profile of patients with colorectal cancer and polyps. METHODS: Patients with stage I-III CRC, adenomatous polyps and individuals with normal colonoscopy were selected. All patients underwent peripheral blood collection for lipid extraction. The samples were analyzed by MALDI-MS technique for lipid identification. STATISTICAL ANALYSIS: Univariate and multivariate (principal component analysis [PCA] and discriminant analysis by partial least squares [PLS-DA]) analyses workflows were applied to the dataset, using MetaboAnalyst 3.0 software. The ions were identified according to the class of lipids using the online database Lipid Maps (http://www.lipidmaps.org). RESULTS: We included 88 individuals, 40 with CRC, 12 with polyps and 32 controls. Boxplot analysis showed eight VIP ions in the three groups. Differences were observed between the cancer and control groups, as well as between cancer and polyp, but not between polyps and control. The polyketide (810.1) was the lipid represented in cancer and overrepresented in polyp and control. Among the patients with CRC we observed differences between lipids with lymph node invasion (N1-2) compared to those without lymph node invasion (N). CONCLUSION: Possible lipid biomarkers were identified among cancer patients compared to control and polyp groups. The polyketide lipid (810.1) was the best biomarker to differentiate the cancer group from control and polyp. We found no difference between the biomarkers in the polyp group in relation to the control.

Multiple sclerosis has a distinct lipid signature in plasma and cerebrospinal fluid.

OBJECTIVE: The diagnosis of multiple sclerosis (MS) has changed over the last decade, but remains a composite of clinical assessment and magnetic resonance imaging to prove dissemination of lesions in time and space. The intrathecal synthesis of immunoglobulin may be a nonspecific marker and there are no plasma biomarkers that are useful in the diagnosis of MS, presenting additional challenges to their early detection. METHODS: We performed a preliminary untargeted qualitative lipidomics mass spectrometry analysis, comparing cerebrospinal fluid (CSF) and plasma samples from patients with MS, other inflammatory neurological diseases and idiopathic intracranial hypertension. RESULTS: Lipid identification revealed that fatty acids and sphingolipids were the most abundant classes of lipids in the CSF and that glycerolipids and fatty acids were the main class of lipids in the plasma of patients with MS. The area under the curve was 0.995 (0.912-1) and 0.78 (0.583-0.917), respectively. The permutation test indicated that this ion combination was useful for distinguishing MS from other inflammatory diseases (p < 0.001 and 0.055, respectively). CONCLUSION: This study concluded that the CSF and plasma from patients with MS bear a unique lipid signature that can be useful as a diagnostic biomarker.

Expression of tumor suppressors miR-195 and let-7a as potential biomarkers of invasive breast cancer.

OBJECTIVE: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at the posttranscriptional level. Some miRNAs, including let-7a and miR-195, have been described as tumor suppressors. However, the roles of these microRNAs in breast cancer progression remain controversial. The aim of this study is to evaluate miR-195 and let-7a expression as potential biomarkers of invasive breast cancer. METHODS: In the present study, 200 individuals were separated into three groups: (i) 72 women constituting the control group who were selected according to rigorous and well-established criteria; (ii) 56 patients with benign breast tumors; and (iii) 72 patients with malignant breast cancers of different clinical stages. The miR-195 and let-7a expression levels in serum were evaluated by real-time PCR. The results were assessed alone and in combination, and the analysis included an estimation of sensitivity and specificity in ROC curves. RESULTS: Compared with the benign and control groups, both microRNAs were downregulated in the malignant breast cancer patient group. Compared with the malignant group, the combination of both biomarkers in the control and benign groups showed good sensitivity and specificity in the serum with AUCs of 0.75 and 0.72, respectively. The biomarker combination for the control group versus the malignant group exhibited a better sensitivity and specificity than for the benign group versus the malignant group. CONCLUSION: These findings support the evidence that the analysis of miR-195 and let-7a can be used as a non-invasive biomarker for breast cancer detection.

Omega-3 PUFA modulate lipogenesis, ER stress, and mitochondrial dysfunction markers in NASH - Proteomic and lipidomic insight.

BACKGROUND & AIMS: Currently there is no FDA-approved therapy for nonalcoholic steatohepatitis (NASH). Increased n-6/n-3 polyunsaturated fatty acids (PUFA) ratio can induce endoplasmic reticulum (ER) stress and mitochondrial dysfunction that characterize NASH. Our recent study with n-3 PUFA showed improvement in individual histologic parameters like steatosis, ballooning and lobular inflammation. We hypothesized that n-3 PUFA therapy mediated improvement in histologic parameters is modulated by lipidomic and proteomic changes. METHODS: We therefore evaluated hepatic proteomic and plasma lipidomic profiles before and after n-3 PUFA therapy in subjects with NASH. In a double-blind, randomized, placebo-controlled trial, patients with NASH received 6-month treatment with n-3 PUFA (0.945 g/day [64% alpha-linolenic (ALA), 21% eicosapentaenoic (EPA), and 16% docosahexaenoic (DHA) acids]). Paired liver biopsy and plasma collected before and after-n-3 PUFA therapy were assessed using mass spectrometry and gas chromatography for hepatic proteomics and plasma lipidomics. Data were matched to UniProt and LIPID MAPS database, respectively. Cytoscape software was used to analyze functional pathways. Twenty-seven NASH patients with paired liver histology and plasma before and after n-3 PUFA treatment were studied. RESULTS: Treatment with n-3 PUFA significantly increased ALA, EPA, and glycerophospholipids, and decreased arachidonic acid (p < 0.05 for all). Further, proteomic markers of cell matrix, lipid metabolism, ER stress and cellular respiratory pathways were also modulated. Interestingly, these alterations reflected functional changes highly suggestive of decreased cellular lipotoxicity potential; reduced ER proteasome degradation of proteins and induction of chaperones; and a shift in cell energy homeostasis towards mitochondrial beta-oxidation. CONCLUSION: Six-month treatment with omega-3 PUFAs significantly improved hepatic proteomic and plasma lipidomic markers of lipogenesis, endoplasmic reticulum stress and mitochondrial functions in patients with NASH.

Association between the p53 arginine/arginine homozygous genotype at codon 72 and human papillomavirus E6/E7 mRNA expression.

OBJECTIVE: To evaluate the association between p53 polymorphisms and human papillomavirus (HPV) E6/E7 mRNA expression. METHODS: We analyzed 175 cervical samples from women aged 16-69 years old who were tested for HPV E6/E7 mRNA expression (NucliSENS® EasyQ® HPV). The samples were divided into three groups: positive (n=75) those with positive HPV E6/E7 mRNA expression and positive high-risk HPV Hybrid Capture (HR-HC) test; negative (n=52) those with negative HPV E6/E7 mRNA expression and positive HR-HC; and control (n=48) those with negative HPV E6/E7 mRNA expression and negative HR-HC. The p53 polymorphisms at codons 11, 72, and 248 were evaluated through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The frequency of the arginine/arginine homozygous genotype at codon 72 was significantly higher in the positive (49.3%) than in the negative (32.7%) and control groups (20.8%, p=0.002*). The frequency of the arginine allele was also significantly higher in the positive (67.3%) than in the negative (53.8%) and control groups (38.5%, p<0.001*). The arginine/arginine homozygous genotype was significantly associated with positive HPV E6/E7 mRNA expression (positive group) compared with negative and control groups (odds ratio: 2.633; 95% CI, 1.399-4.954, p=0.003). The frequency of arginine/arginine homozygous genotype at codon 72 remained significantly more frequent in the positive group of women aged ≥30 years than in the other two groups. CONCLUSION: The presence of the p53 arginine/arginine homozygous genotype at codon 72 was significantly associated with the positive HPV E6/E7 mRNA expression.

[Role of the genetic polymorphism of p53 (codon 72) gene in colorectal cancer]. / Estudo do polimorfismo genético no gene p53 (códon 72) em câncer colorretal.

BACKGROUND: Polymorphisms are genetic variations that can occur in sequences of codons, leading to defective proteins. p53 is the most commonly gene affected in human cancer. The polymorphism of this gene occurs by a substitution of a base in codon 72 and may increase the risk of cancer. AIM: To investigate the possible association between p53 arginine/72 proline polymorphism and susceptibility to colorectal cancer. PATIENTS AND METHODS: This polymorphism was studied by polymerization chain reaction using specific primers in 100 patients with colorectal cancer paired by sex and age to 100 patients without cancer. Alcohol and tobacco used by all the patients and clinical aspects as stage, grade of differentiation and recurrence in the case group was compared with the genotype analyzed. RESULTS: The frequency of homozygosis for arginine was 56% in the cancer group and 58% in the control group. No significant difference was observed among both groups. This genotype was more frequent in colorectal cancer patients stage IV than in stage I (80% versus 14%). There was no significant difference between genotypes and alcohol, tobacco, grade of differentiation or recurrence. CONCLUSION: Homozygosity for arginine was the most prevalent genotype in both groups. The frequency of codon 72 proline/arginine p53 gene polymorphism was not correlated with a higher risk of colorectal cancer. Arginine/arginine genotype was more prevalent in advanced cancer patients (stage IV).

Estudo de biomarcadores lipídicos em pacientes portadores com pólipos e câncer colorretal / Study of lipid biomarkers of patients with polyps and colorectal câncer

ABSTRACT BACKGROUND: Colorectal cancer (CRC) is one of the leading causes of cancer worldwide. Early diagnostic methods using serum biomarkers are required. The study of omics, most recently lipidomics, has the purpose of analyzing lipids for a better understanding of human lipidoma. The evolution of mass spectrometry methods, such as MALDI-MS technology, has enabled the detection and identification of a wide variety of lipids with great potential to open new avenues for predictive and preventive medicine. OBJECTIVE: To determine the lipid profile of patients with colorectal cancer and polyps. METHODS: Patients with stage I-III CRC, adenomatous polyps and individuals with normal colonoscopy were selected. All patients underwent peripheral blood collection for lipid extraction. The samples were analyzed by MALDI-MS technique for lipid identification. STATISTICAL ANALYSIS: Univariate and multivariate (principal component analysis [PCA] and discriminant analysis by partial least squares [PLS-DA]) analyses workflows were applied to the dataset, using MetaboAnalyst 3.0 software. The ions were identified according to the class of lipids using the online database Lipid Maps (http://www.lipidmaps.org). RESULTS: We included 88 individuals, 40 with CRC, 12 with polyps and 32 controls. Boxplot analysis showed eight VIP ions in the three groups. Differences were observed between the cancer and control groups, as well as between cancer and polyp, but not between polyps and control. The polyketide (810.1) was the lipid represented in cancer and overrepresented in polyp and control. Among the patients with CRC we observed differences between lipids with lymph node invasion (N1-2) compared to those without lymph node invasion (N). CONCLUSION: Possible lipid biomarkers were identified among cancer patients compared to control and polyp groups. The polyketide lipid (810.1) was the best biomarker to differentiate the cancer group from control and polyp. We found no difference between the biomarkers in the polyp group in relation to the control.

RESUMO CONTEXTO: O câncer colorretal (CCR) é, mundialmente, uma das principais causas de câncer. Métodos de diagnóstico precoce através de biomarcadores séricos são necessários. O estudo das ômicas, mais recentemente a lipidômica, tem a finalidade de analisar os lipídeos para melhor compreensão do lipidoma humano. A evolução dos métodos de espectrometria de massa, como a tecnologia por MALDI-MS, possibilitou a detecção e a identificação de uma ampla variedade de lipídeos com grande potencial para abrir novos caminhos para a medicina preditiva e preventiva. OBJETIVO: Determinar o perfil lipidômico de pacientes com câncer colorretal e pólipos. MÉTODOS: Foram selecionados pacientes com CCR estádio I-III, com pólipos adenomatosos e indivíduos com colonoscopia normal. Todos os pacientes foram submetidos a coleta do sangue periférico para extração do lipídeo. As amostras foram analisadas por técnica de MALDI-MS para a identificação dos lipídeos. ANÁLISE ESTATÍSTICA: Para análise univariada e multivariada foram utilizados a análise de componentes principais (PCA) e a análise discriminante pelos quadrados mínimos (PLS-DA). Os íons foram identificados de acordo com a classe de lipídeos usando-se o Lipid Maps (http://www.lipidmaps.org). RESULTADOS: Foram incluídos 88 indivíduos, 40 com CCR, 12 com pólipos e 32 controles. A análise de boxbolt evidenciou oito íons VIP nos três grupos. Observou-se diferenças entre os grupos câncer e controle, assim como entre câncer e pólipo, mas não entre pólipos e controle. O policetídeo (810,1) foi o lipídeo hipo-representado no câncer e hiperrepresentado no pólipo e controle. Entre os pacientes com CCR observamos diferenças entre os lipídeos com invasão linfonodal (N1-2) comparados aos sem invasão linfonodal (N0). CONCLUSÃO: Foram identificados possíveis biomarcadores lipídicos entre os pacientes com câncer comparados aos grupos controle e pólipo. O lipídeo policetídeo (810,1) foi o melhor biomarcador para diferenciar o grupo câncer do controle e pólipo. Não encontramos diferença entre os biomarcadores no grupo pólipo em relação ao controle.

Prevalence of sexually transmitted diseases in female athletes in São Paulo, Brazil.

OBJECTIVE: To determine the prevalence of sexually transmitted diseases in female athletes. METHODS: An observational, cross-sectional study was conducted including 50 female athletes with mean age of 20 ± 3 years. Colposcopy, pap smear, and polymerase chain reaction for Chlamydia trachomatis, human papillomavirus and Neisseria gonorrhoeae were performed. Blood samples were collected to test for the human immunodeficiency virus, syphilis, hepatitis B and C. The athletes presenting clinical diseases or conditions identifiable by laboratory tests were treated and followed up in the unit. RESULTS: Forty-six percent of the participants were unaware of sexually transmitted diseases. The prevalence of sexually transmitted diseases among athletes was 48% (24 cases). Human papillomavirus was the most frequent agent (44%). Considering the human papillomavirus genotypes, subtype 16 was the most prevalent (53%), followed by 11-6 (22%) and 18 (13%). Two athletes tested positive for C. trachomatis. There were no cases diagnosed of infection by N. gonorrhoeae, syphilis, hepatitis B, hepatitis C and human immunodeficiency virus. However, only 26 athletes had been vaccinated for hepatitis B. CONCLUSION: The prevalence of sexually transmitted diseases in female athletes was high. Primary prevention measures (hepatitis B and human papillomavirus vaccination) and secondary (serology, pap smears) must be offered to this specific group of women. The matter should be further approached in sports.

Multiple sclerosis has a distinct lipid signature in plasma and cerebrospinal fluid / Esclerose múltipla tem uma assinatura lipídica distinta no plasma e no líquido cefalorraquiano

ABSTRACT The diagnosis of multiple sclerosis (MS) has changed over the last decade, but remains a composite of clinical assessment and magnetic resonance imaging to prove dissemination of lesions in time and space. The intrathecal synthesis of immunoglobulin may be a nonspecific marker and there are no plasma biomarkers that are useful in the diagnosis of MS, presenting additional challenges to their early detection. Methods We performed a preliminary untargeted qualitative lipidomics mass spectrometry analysis, comparing cerebrospinal fluid (CSF) and plasma samples from patients with MS, other inflammatory neurological diseases and idiopathic intracranial hypertension. Results Lipid identification revealed that fatty acids and sphingolipids were the most abundant classes of lipids in the CSF and that glycerolipids and fatty acids were the main class of lipids in the plasma of patients with MS. The area under the curve was 0.995 (0.912-1) and 0.78 (0.583-0.917), respectively. The permutation test indicated that this ion combination was useful for distinguishing MS from other inflammatory diseases (p < 0.001 and 0.055, respectively). Conclusion This study concluded that the CSF and plasma from patients with MS bear a unique lipid signature that can be useful as a diagnostic biomarker.

RESUMO Embora o diagnóstico da EM tenha se modificado na última década, ainda tem como requisito básico a demonstração da disseminação no tempo e no espaço, através do quadro clínico e do exame de ressonância magnética. A síntese intratecal de imunoglobulina pode ser um marcador inespecífico e não há biomarcadores plasmáticos que sejam úteis no diagnóstico da EM, impondo desafios à sua detecção precoce. Métodos Realizamos uma análise lipidômica preliminar por espectrometria de massas, não direcionada, qualitativa, comparando amostras de LCR e plasma de pacientes com EM, outras doenças neurológicas inflamatórias e hipertensão intracraniana idiopática (HII). Resultados A identificação lipídica revelou que os ácidos graxos e esfingolipídios foram as classes mais abundantes de lipídios no LCR e que glicerolipídios e ácidos graxos foram a principal classe de lipídios no plasma de pacientes com EM. A AUC foi de 0,995 (0,912-1) e 0,78 (0,583-0,917), respectivamente. O teste de permutação indicou que essa combinação de íons foi útil para distinguir a EM de outras doenças inflamatórias (p < 0,001 e 0,055, respectivamente). Conclusão Este estudo sugere que o líquido cefalorraquidiano (LCR) e o plasma de pacientes com EM possuem uma assinatura lipídica única, pode ser útil como um biomarcador diagnóstico.

Expression of tumor suppressors miR-195 and let-7a as potential biomarkers of invasive breast cancer

OBJECTIVE: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at the posttranscriptional level. Some miRNAs, including let-7a and miR-195, have been described as tumor suppressors. However, the roles of these microRNAs in breast cancer progression remain controversial. The aim of this study is to evaluate miR-195 and let-7a expression as potential biomarkers of invasive breast cancer. METHODS: In the present study, 200 individuals were separated into three groups: (i) 72 women constituting the control group who were selected according to rigorous and well-established criteria; (ii) 56 patients with benign breast tumors; and (iii) 72 patients with malignant breast cancers of different clinical stages. The miR-195 and let-7a expression levels in serum were evaluated by real-time PCR. The results were assessed alone and in combination, and the analysis included an estimation of sensitivity and specificity in ROC curves. RESULTS: Compared with the benign and control groups, both microRNAs were downregulated in the malignant breast cancer patient group. Compared with the malignant group, the combination of both biomarkers in the control and benign groups showed good sensitivity and specificity in the serum with AUCs of 0.75 and 0.72, respectively. The biomarker combination for the control group versus the malignant group exhibited a better sensitivity and specificity than for the benign group versus the malignant group. CONCLUSION: These findings support the evidence that the analysis of miR-195 and let-7a can be used as a non-invasive biomarker for breast cancer detection.

Association between the p53 arginine/arginine homozygous genotype at codon 72 and human papillomavirus E6/E7 mRNA expression

ABSTRACT Objective: To evaluate the association between p53 polymorphisms and human papillomavirus (HPV) E6/E7 mRNA expression. Methods: We analyzed 175 cervical samples from women aged 16-69 years old who were tested for HPV E6/E7 mRNA expression (NucliSENS® EasyQ® HPV). The samples were divided into three groups: positive (n = 75) those with positive HPV E6/E7 mRNA expression and positive high-risk HPV Hybrid Capture (HR-HC) test; negative (n = 52) those with negative HPV E6/E7 mRNA expression and positive HR-HC; and control (n = 48) those with negative HPV E6/E7 mRNA expression and negative HR-HC. The p53 polymorphisms at codons 11, 72, and 248 were evaluated through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequency of the arginine/arginine homozygous genotype at codon 72 was significantly higher in the positive (49.3%) than in the negative (32.7%) and control groups (20.8%, p = 0.002*). The frequency of the arginine allele was also significantly higher in the positive (67.3%) than in the negative (53.8%) and control groups (38.5%, p < 0.001*). The arginine/arginine homozygous genotype was significantly associated with positive HPV E6/E7 mRNA expression (positive group) compared with negative and control groups (odds ratio: 2.633; 95% CI, 1.399-4.954, p = 0.003). The frequency of arginine/arginine homozygous genotype at codon 72 remained significantly more frequent in the positive group of women aged ≥30 years than in the other two groups. Conclusion: The presence of the p53 arginine/arginine homozygous genotype at codon 72 was significantly associated with the positive HPV E6/E7 mRNA expression.

Assessment of glutathione S-transferaseM1 (GSTM1) and its polymorphisms GSTM1 null in the response to treatment with chemotherapy in advanced ovarian carcinoma.

OBJECTIVE: To assess if the genotype of the glutathione S-transferase M1 (GSTM1) enzyme and its GSTM1 null polymorphism can influence the response to chemotherapeutic treatment of advanced ovarian cancer. METHODS: Case-control study of 112 patients with advanced ovarian cancer submitted to chemotherapy during the period from January 1995 to December 2005. The tissue to study the GSTM1 genotype and its deletion came from surgical staging to treat ovarian cancer. The PCR product generates two distinct genotypes, characterized as positive and null. The response to chemotherapy was evaluated using the World Health Organization (WHO) criteria. Patients were classified as having: a) no response, b) a response. RESULTS: The presence of GSTM1 or its GSTM1 null polymorphism did not influence the preoperative chemotherapy response. Among the patients who did respond, 88.9% presented with positive GSTM1 and 11.1% with null GSTM1. Among the patients that did not respond, 85.71% presented with positive GSTM1 and 14.29% with null GSTM1 (p = 0.825). GSTM1 and its GSTM1 null polymorphism had no influence on the postoperative response to chemotherapy. Among the patients who did respond, 80.65% presented with positive GSTM1 and 19.35% with null GSTM1. Among the patients who did not respond, 87.50% presented with positive GSTM1 and 12.5% with the null polymorphism (p = 0.553). CONCLUSION: No difference was observed in the response to treatment with chemotherapy in patients with advanced ovarian cancer, as to the GSTM1 genotype compared to its GSTM1 null polymorphism.

High Risk HPV E6/E7 Oncoprotein Expression in Women with High Grade Squamous Intraepithelial Lesion / Expressão de oncoproteinas E6/e7 do HPV de alto risco em Mulheres com Lesão Intraepitelial Escamosa de alto grau

Purpose To correlate the expression of high-risk HPV E6 mRNA with pap smear, colposcopy, and biopsy results in women with high grade squamous intraepithelial lesion (HSIL). Methods A cross-sectional study was performed on women referred for primary care services after cytological diagnosis of HSIL. We evaluated the expression of E6/E7 mRNA of HPV types 16,18,31,33, and 45 and correlated the results with those of Pap smear, colposcopy, and biopsy. For amplification/detection of mRNA E6 / E7 we used NucliSENSEasyQ kit to detect HPV mRNA by polymerase chain reaction with primers/ probes for HPV types 16, 18, 31, 33, and 45. Results Out of 128 valid tests, the results of 30 (23.4%) tests were negative and 98 (70%) tests were positive. Only one type of HPV was detected in 87.7% of the E6/E7 mRNA positive cases. HPV16 was detected in 61.2% of the cases, followed by HPV33 (26.5%), HPV31 (17.3%), HPV18 (10%), and HPV45 (4.08%). Pap smear tests revealed that the E6/E7 test was positive in 107 (83.8%) women with atypical squamous cells - high grade (ASC-H), HSIL, or higher. The E6/E7 test was positive in 69 (57.5%) specimens presenting negative cytology results. When analyzing the association with colposcopy results, the frequency of positive E6/E7 results increased with the severity of the injury, ranging from 57.1% in women without colposcopy-detected injury to 86.5% in those with higher levels of colposcopy findings. Of the 111 women who underwent biopsy and E6/E7 testing, the E6/E7 test was positive in 84.7% of the women who presented with lesions of cervical intraepithelial neoplasia (CIN) grade 2 or higher. Finally, 41.2% of women with a negative biopsy presented a positive E6/E7 test. Conclusions E6/E7mRNA expression was higher in women with HSIL and CIN grade 2 or higher.

Objetivo Correlacionar a expressão mRNAE6/E7 do HPV de alto risco com os exames de Papanicolau, colposcopia e biópsia em mulheres com lesão intraepitelial escamosa de alto grau (HSIL). Métodos Estudo transversal com mulheres encaminhadas aos serviços de atenção primária com diagnóstico citológico de HSIL. Foi avaliada a expressão do mRNAE6/E7 dos tipos de HPV 16,18,31,33 e 45, correlacionando-se a expressão com os exames de Papanicolau, colposcopia e biópsia. Para a amplificação/detecção demRNA de E6/E7 foi usado o kit NucliSENS EasyQ(r) HPV que detecta mRNA do HPV por meio da reação em cadeia da polimerase com primers/probes HPV dos tipos 16, 18, 31, 33 e 45. Resultados Foram obtidos 128 testes válidos. Destes: 30 (23,4%) foram negativos e 98 (70%) dos testes foram positivos. Foi encontrado apenas um tipo de HPV em 87,7% dos positivos. O HPV16 foi omais encontrado em61,2%, seguido pelos HPV33 (26,5%); HPV31 (17,34%); HPV 18 (10,0%) e HPV (45 4,0%). Quanto ao exame de Papanicolau, o teste E6/E7 foi positivo em 107 (83,8%) das mulheres com ASC-H, HSIL ou superior, enquanto em citologia negativa foi encontrado um resultado positivo em 69 (57,5%) colposcopia. A frequência de teste E6/E7 positivo aumentou com a gravidade da lesão, detectada na colposcopia variando de 57,1% em mulheres sem lesão identificada em colposcopia até 86,5% naqueles com achado de colposcopia de grau maior. Das 111 mulheres que se submeteram a biópsia e o teste E6/E7, o teste foi positivo em 84,7% das que apresentaramlesão igual ou superior a NIC 2 (neoplasia intraepitelial cervical) e 41,2% daqueles com biópsia negativa. Conclusões A expressão de E6, E7 RNAm ocorreu commaior frequência emlesões de alto grau citológica e em casos com biópsias de NIC2 ou maior.

[Endometrial polyps: clinical and epidemiological aspects and analysis of polymorphisms]. / Pólipos endometriais: aspectos clínicos, epidemiológicos e pesquisa de polimorfismos.

PURPOSE: To evaluate the clinical and epidemiological risk factors for endometrial cancer in postmenopausal women with endometrial polyps, as well as the genetic polymorphism of the progesterone receptor (PROGINS). METHODS: A case-control study was designed with 160 postmenopausal women with endometrial polyps, compared to a normal Control Group of 400 postmenopausal women. The genotyping of PROGINS polymorphism was determined by the polymerase chain reaction. Clinical and epidemiological data were compared between benign endometrial polyps and 118 of the control subjects. Variables were also compared with regard to benign and malignant endometrial polyps. RESULTS: Comparison of the epidemiological variables between groups showed a significant difference for age, ethnicity, time since menopause, parity, tamoxifen use, hypertension and breast cancer, all of them more prevalent in the polyp group. After adjustment for age, statistical significance remained only for parity (OR=1.1), hypertension (OR=2.2) and breast cancer (OR=14.4). There were six cases of malignant polyps (3.7%). The frequency of bleeding was 23.4% for benign polyps and 100% for malignant polyps, with large polyps being detected in 54.6% of the benign cases and in 100 of the malignnat ones. The frequency of arterial hypertension was 54.5% for benign polyps and 83.3% for the malignant ones. The frequency of PROGINS T1/T1, T1/T2 and T2/T2 polymorphism was 79.9%, 19.5% and 0.6%, respectively, for the polyp group, and 78.8%, 20.8% and 0.5% for the Control Group. CONCLUSIONS: Elderly age, hypertension, and breast cancer were significantly associated with endometrial polyps. The presence of PROGINS polymorphism was not significantly associated with endometrial polyps. The incidence of malignant polyps was low and strongly associated with bleeding, large-sized polyp and arterial hypertension.

Genetic analysis and cAMP measurement: comparison between lean and obese anovulating mice / Análise genética e dosagem de AMP cíclico: comparação entre camundongos fêmeas anovuladoras magras e obesas

PURPOSE: To evaluate genes differentially expressed in ovaries from lean (wild type) and obese (ob/ob) female mice and cyclic AMP production in both groups. METHODS: The expression on messenger RNA levels of 84 genes concerning obesity was analyzed through the PCR array, and cyclic AMP was quantified by the enzyme immunoassay method. RESULTS: The most downregulated genes in the Obesity Group included adenylate cyclase-activating polypeptide type 1, somatostatin, apolipoprotein A4, pancreatic colipase, and interleukin-1 beta. The mean decrease in expression levels of these genes was around 96, 40, 9, 4.2 and 3.6-fold, respectively. On the other hand, the most upregulated genes in the Obesity Group were receptor (calcitonin) activity-modifying protein 3, peroxisome proliferator activated receptor alpha, calcitonin receptor, and corticotropin-releasing hormone receptor 1. The increase means in the expression levels of such genes were 2.3, 2.7, 4.8 and 6.3-fold, respectively. The ovarian cyclic AMP production was significantly higher in ob/ob female mice (2,229±52 fMol) compared to the Control Group (1,814±45 fMol). CONCLUSIONS: Obese and anovulatory female mice have reduced reproductive hormone levels and altered ovogenesis. Several genes have their expression levels altered when leptin is absent, especially adenylate cyclase-activating polypeptide type 1. .

OBJETIVO: Avaliar os genes diferencialmente expressos em ovários de camundongos fêmeas magras (tipo selvagem) e obesas (ob/ob) e a produção de AMP cíclico em ambos os grupos. MÉTODOS: A expressão nos níveis de RNA mensageiro de 84 genes relacionados à obesidade foi analisada por PCR Array, e o AMP cíclico foi quantificado por método imunoenzimático. RESULTADOS: Os genes que mais sofreram diminuição da expressão no Grupo Obesidade incluíram o tipo 1 de polipeptídeo ativador da adenilato ciclase, o da somatostatina, da apolipoproteína A4, da colipase pancreática e da beta interleucina 1. A média de redução na expressão desses genes foi de aproximadamente 96, 40, 9, 4,2 e 3,6 vezes, respectivamente. Por outro lado, os genes que mais tiveram aumento na expressão no Grupo Obesidade foram o gene da proteína modificadora da atividade do receptor de calcitonina 3, do proliferador de peroxissomos ativados por proteína alfa, do receptor de calcitonina e do receptor para hormônio liberador de corticotropinas 1. As médias de acréscimo nos níveis de expressão de tais genes foram de 2,3, 2,7, 4,8 e 6,3 vezes, respectivamente. A produção de AMP cíclico ovariana foi significantemente aumentada em camundongos fêmeas ob/ob (2.229±52 fMol) quando comparada ao Grupo Controle (1.814±45 fMol). CONCLUSÕES: Camundongos fêmeas obesas e anovuladoras possuem níveis de hormônio reprodutivo reduzidos e ovulogênese alterada. Vários genes mostram níveis de expressão alterados quando a leptina está ausente, principalmente o tipo 1 de polipeptídeo ativador da adenilato ciclase. .

Analysis of GB virus C infection among HIV-HCV coinfected patients.

The aim of this study was to evaluate the effect of GB virus C on laboratory markers and histological parameters among HIV-seropositive patients coinfected with HCV. Lower degrees of hepatic lesions were observed in the triple-infected patients, in comparison with HIV-HCV coinfected patients who were negative for GBV-C RNA.