Is the expansion of the subway network alone capable of improving local air quality? A study case in São Paulo, Brazil.

One of the policies adopted to reduce vehicular emissions is subway network expansion. This work fitted interrupted regression models to investigate the effects of the inauguration of subway stations on the mean, trend, and seasonality of the NO, NO2, NOx, and PM10 local concentrations. The regions investigated in the city of São Paulo (Brazil) were Pinheiros, Butantã, and St. Amaro. In Pinheiros, after the inauguration of the subway station, there were downward trends for all pollutants. However, these trends were not significantly different from the trends observed before. In Butantã, only regarding NO, there was a significant reduction and seasonal change after the subway station's inauguration. In St. Amaro, no trend in the PM10 concentration was noted. The absence of other transportation and land use policies in an integrative way to the subway network expansion may be responsible for the low air quality improvement. This study highlights that the expansion of the subway network must be integrated with other policies to improve local air quality.

Multimorbidities Are Associated to Lower Survival in Ischaemic Stroke: Results from a Brazilian Stroke Cohort (EMMA Study).

BACKGROUND: Stroke prognosis is related to the multimorbidity profile. Moreover, performing an individual evaluation of most common cerebrovascular risk factors (CVRF) not always identifies patients with poor prognosis. Thus, we decided to evaluate multimorbidity profile, focusing on the Charlson Comorbidity Index (CCI) validated by Goldstein for ischaemic stroke (IS) patients, a score that measures a burden of comorbidities and its related mortality in the long-term survival of the EMMA Study (Study of Stroke Mortality and Morbidity). METHODS: Nine hundred fifty-nine individuals (median age 70 years) had validated data on the diagnosis of IS, main CVRF and clinical comorbidities pre index event such as atrial fibrillation (AF), stroke recurrence, diabetes, hypertension, heart failure and cancer. CCI modified by Goldstein was calculated, which includes 17 clinical conditions with scores ranging from 1 to 6 (0-31 points). Survival analyses were performed by Kaplan-Meier curves and Cox logistic regression models (cumulative hazard ratio [HR] with [95% CI]) for all-cause mortality at 180 days, and every 3 years up to 9-year follow-up. Mortality analyzes were performed by CCI categorized according to weight added to comorbidities (Reference group: zero, moderate: 1, severe: 2 and very severe: ≥3 points). We also tested the modification effect of AF and stroke recurrence including these conditions in the CCI. RESULTS: The overall survival rate was 47% (508 deaths/959). The worst survival (577, 95% CI 381-773 days) and the highest risk of death after stroke were observed in the very severe CCI group (HR 3.18; 95% CI 2.16-4.69) up to 9 years. The inclusion of previous AF and stroke in the CCI slightly increased the risk of death for very severe CCI (HR 3.27; 95% CI 2.07-5.18). CONCLUSIONS: A high burden of comorbidities represented an independent predictor of poor prognosis increasing the risk of dying by 2 to 3 times among IS up to 9 years in the EMMA study. The inclusion of other CVRF such as AF and stroke recurrence slightly modified all-cause mortality risk.

Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with anastrozole.

OBJECTIVES: To evaluate Ki-67 antigen expression in mammary epithelium of female rats in persistent estrus treated with anastrozole. MATERIALS AND METHODS: Twenty-eight Wistar-Hanover female rats in persistent estrus induced by subcutaneous injection of 1.25 mg of testosterone propionate in the second day of life were randomly divided into two groups, control and experimental, with 14 animals each. The animals of control group received only the vehicle (propyleneglycol) and the animals of group experimental received 0.125 mg daily of anastrozole by gavage during 28 days. After 28 days of treatment, all animals were sacrificed and the first pair of abdominal-inguinal mammary glands was removed and fixed in 10% buffered formalin to investigate Ki-67 antigen expression by immunohistochemistry. RESULTS: The mean percentage of Ki-67-stained nuclei per 500 cells in the mammary epithelium was 76.97 ± 0.76 and 14.44 ± 2.02 [mean ± standard error of the mean (SEM)] in the control and experimental groups, respectively (p < 0.0001). CONCLUSIONS: Anastrozole treatment significantly reduced Ki-67 expression in the mammary epithelium of rats in persistent estrus.

Major Depression as a Predictor of Poor Long-Term Survival in a Brazilian Stroke Cohort (Study of Stroke Mortality and Morbidity in Adults) EMMA study.

BACKGROUND: The influence of poststroke depression on long-term survival is poorly investigated. Thus, we aimed to evaluate the influence of major depression disorder (MDD) on long-term survival in the participants from The Study of Stroke Mortality and Morbidity in Adults (EMMA Study) in São Paulo, Brazil. METHODS: We prospectively evaluated ischemic and hemorrhagic stroke (HS) cases from the EMMA Study. Baseline and stroke characteristics and cardiovascular risk factors were evaluated according to MDD assessed by the Patient Health Questionnaire, which was applied 30 days after index event and periodically during 1-year follow-up. We performed Kaplan-Meier survival analysis, as well as crude and multiple Cox proportional hazards models. RESULTS: In this subset of the EMMA Study, we evaluated 164 (85.9%) patients with ischemic stroke and 27 (14.1%) with HS. Among these, overall incidence of MDD was 25.1% during 1 year of follow-up, regardless stroke subtype. The peak rate of major depression postacute event was beyond 1 month. We observed a lower survival rate among individuals who developed poststroke MDD than among those who did not develop this condition after 1 year of follow-up (85.4% versus 96.5%, log rank P = .006). After multiple analysis, we kept a higher risk of all-cause mortality among those who developed MDD compared to participants without MDD (hazard ratio = 4.60, 95% confidence interval = 1.36-15.55, P = .01). CONCLUSION: Our findings suggest that incident MDD is a potential marker of poor prognosis 1 year after stroke.

Stroke in the rain forest: prevalence in a ribeirinha community and an urban population in the Brazilian Amazon.

BACKGROUND: Our objective was to determine the cerebrovascular prevalence in a town in the Brazilian Amazon basin and compare the ribeirinhos (riparians) to the urban population in the same municipality. METHODS: From May to October 2011, 6,216 residents over 35 years of age in the town of Coari were interviewed using a screening questionnaire, the Stroke Symptom Questionnaire. Cerebrovascular prevalence rates (PRs) from the door-to-door surveillance were calculated according to the location of the home. RESULTS: Respondent totals were 4,897 in the urban area and 1,028 in the rural area. The crude prevalence of stroke was 6.3% in rural and 3.7% in urban areas with differences maintained after sex and age adjustment. Among stroke cases, the ribeirinhos were those with less access to medical care in comparison to the urban area (32.1 vs. 52.5%, p = 0.01), and a positive association between rural area and no medical care for stroke remained (PR, 1.33; 95% confidence interval, 1.03-1.71), independently of age, sex, education and functional impairment. CONCLUSIONS: This study provides the first population-based cerebrovascular prevalence comparison between an urban and a rural population in the Amazon rain forest. The PRs were higher in the ribeirinha compared to the urban population in the same municipality.

Effect of 10-valent pneumococcal vaccine on pneumonia among children, Brazil.

Pneumonia is most problematic for children in developing countries. In 2010, Brazil introduced a 10-valent pneumococcal conjugate vaccine (PCV10) to its National Immunization Program. To assess the vaccine's effectiveness for preventing pneumonia, we analyzed rates of hospitalization among children 2-24 months of age who had pneumonia from all causes from January 2005 through August 2011. We used data from the National Hospitalization Information System to conduct an interrupted time-series analysis for 5 cities in Brazil that had good data quality and high PCV10 vaccination coverage. Of the 197,975 hospitalizations analyzed, 30% were for pneumonia. Significant declines in hospitalizations for pneumonia were noted in Belo Horizonte (28.7%), Curitiba (23.3%), and Recife (27.4%) but not in São Paulo and Porto Alegre. However, in the latter 2 cities, vaccination coverage was less than that in the former 3. Overall, 1 year after introduction of PCV10, hospitalizations of children for pneumonia were reduced.

Ki-67 and Bcl-2 antigen expression in adenomatous colorectal polyps from women with breast cancer.

BACKGROUND: The aim of this study was to evaluate Ki-67 and Bcl-2 antigen expression in colorectal polyps from women with breast cancer. METHODS: A randomized, controlled study was carried out in 35 women, either with or without breast cancer, who had adenomatous colorectal polyps. The patients were divided into two groups: group A (without breast cancer; control group; n = 17) and group B (with breast cancer; study group; n = 18). Immunohistochemistry was performed on the colorectal polyps to evaluate Ki-67 and Bcl-2 antigen expression. Student's t-test and the chi(2) test were used for the statistical analysis of Ki-67 and Bcl-2 expression, respectively. Statistical significance was established as P < 0.05. RESULTS: The mean percentage of Ki-67-stained nuclei in groups A and B was 36.25 +/- 2.31 and 59.44 +/- 3.34 (+/- SEM), respectively (P < 0.0001), while the percentage of cases with cells expressing Bcl-2 in groups A and B was 23.5 and 77.8%, respectively (P < 0.001). CONCLUSIONS: In the present study, there was greater proliferative activity and greater expression of the antiapoptotic protein Bcl-2 in the colorectal polyps of women with breast cancer.

Correction: Mortality among blood donors seropositive and seronegative for Chagas disease (1996-2000) in São Paulo, Brazil: A death certificate linkage study.

[This corrects the article DOI: 10.1371/journal.pntd.0005542.].

Beta-2 adrenergic receptor gene polymorphisms Gln27Glu, Arg16Gly in patients with heart failure.

BACKGROUND: Beta-2 adrenergic receptor gene polymorphisms Gln27Glu, Arg16Gly and Thr164Ile were suggested to have an effect in heart failure. We evaluated these polymorphisms relative to clinical characteristics and prognosis of alarge cohort of patients with heart failure of different etiologies. METHODS: We studied 501 patients with heart failure of different etiologies. Mean age was 58 years (standard deviation 14.4 years), 298 (60%) were men. Polymorphisms were identified by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: During the mean follow-up of 12.6 months (standard deviation 10.3 months), 188 (38%) patients died. Distribution of genotypes of polymorphism Arg16Gly was different relative to body mass index (chi2 = 9.797;p = 0.04). Overall the probability of survival was not significantly predicted by genotypes of Gln27Glu, Arg16Gly, or Thr164Ile. Allele and haplotype analysis also did not disclose any significant difference regarding mortality. Exploratory analysis through classification trees pointed towards a potential association between the Gln27Glu polymorphism and mortality in older individuals. CONCLUSION: In this study sample, we were not able to demonstrate an overall influence of polymorphisms Gln27Glu and Arg16Gly of beta-2 receptor gene on prognosis. Nevertheless, Gln27Glu polymorphism may have a potential predictive value in older individuals.

Effects of raloxifene on Ki-67 and CD34 antigen expression in breast cancer.

BACKGROUND: The aim of this study was to evaluate the effect of raloxifene on CD34 and Ki-67 antigen expression in breast cancer specimens from postmenopausal women. METHODS: Sixteen postmenopausal patients with operable, stage II (>or=3 cm), estrogen receptor-positive breast cancer, who took 60 mg of raloxifene daily for 28 days, participated in this study. Immunohistochemistry was carried out in tumor samples prior to and following raloxifene treatment to evaluate CD34 and Ki-67 protein expression. Angiogenesis was quantified in 10 randomly selected fields per slide, and Ki-67-stained nuclei were counted in 1,000 cells per slide using an image capture and analysis system with 400x magnification. Student's t test for paired samples was used for the statistical analysis of data. Statistical significance was established at p < 0.05. RESULTS: The mean number of microvessels was 44.44 +/- 3.54 prior to raloxifene therapy and 22.63 +/- 1.61 following therapy (p < 0.001), and the mean percentage of Ki-67-stained nuclei was 19.28 +/- 1.61 and 12.13 +/- 1.48 prior to and following raloxifene treatment, respectively (p < 0.001). CONCLUSION: Raloxifene significantly reduces CD34 and Ki-67 protein expression in breast carcinoma in postmenopausal women.

Effectiveness of the Mobile Emergency Medical Services (SAMU): use of interrupted time series.

OBJECTIVE: To evaluate the performance of the Mobile Emergency Medical Services (SAMU) in the ABC Region, using myocardial infarction as tracer condition. METHODS: The analysis of interrupted time series was the approach chosen to test immediate and gradual effects of the intervention on the study population. The research comprised adjusted monthly time series of the hospital mortality rate by myocardial infarction in the period between 2000 and 2011. Data were extracted from the Mortality Information System (SIM), using segmented regression analysis to evaluate the level and trend of the intervention before and after its implementation. To strengthen the internal validity of the study, a control region was included. RESULTS: The analysis of interrupted time series showed a reduction of 0.04 deaths per 100,000 inhabitants in the mortality rate compared to the underlying trend since the implementation of the Emergency Medical Services (p = 0.0040; 95%CI: -0.0816 - -0.0162) and a reduction in the level of 2.89 deaths per 100,000 inhabitants (p = 0.0001; 95%CI: -4.3293 - -1.4623), both with statistical significance. Regarding the control region, Baixada Santista, the difference in the result trend between intervention outcome and post-intervention control of -0.0639 deaths per 100,000 inhabitants was statistically significant (p = 0.0031; 95%CI: -0.1060 - -0.0219). We cannot exclude confounders, but we limited their presence in the study by including control region series. CONCLUSIONS: Although the analysis of interrupted time series has limitations, this modeling can be useful for analyzing the performance of policies and programs. Even though the intervention studied is not a condition that in itself implies effectiveness, the latter would not be present without the former, which, integrated with other conditions, generates a positive result. SAMU is a strategy that must be expanded when formulating and consolidating policies focusing on emergency care.

A non-linear mathematical model using optical sensor to predict heart decellularization efficacy.

One of the main problems of the decellularization technique is the subjectivity of the final evaluation of its efficacy in individual organs. This problem can result in restricted cell repopulation reproducibility and worse responses to transplant tissues. Our proposal is to analyze the optical profiles produced by hearts during perfusion decellularization, as an additional method for evaluating the decellularization process of each individual organ. An apparatus comprised of a structured LED source and photo detector on an adjustable base was developed to capture the relationship between transmitted light during the perfusion of murine hearts, and residual DNA content. Voltage-time graphic records were used to identify a nonlinear mathematical model to discriminate between decellularizations with remaining DNA above (Incomplete Decellularization) and below (Complete Decellularization) the standardized limits. The results indicate that temporal optical evaluation of the process enables inefficient cell removal to be predicted in the initial stages, regardless of the apparent transparency of the organ. Our open system also creates new possibilities to add distinct photo detectors, such as for specific wavelengths, image acquisition, and physical-chemical evaluation of the scaffold, in order to collect different kinds of information, from dozens of studies. These data, when compiled and submitted to machine learning techniques, have the potential to initiate an exponential advance in tissue bioengineering research.

Morphometric analysis of the urethra of castrated female rats treated with tamoxifen.

OBJECTIVES: The aim of this study was to evaluate the effects of tamoxifen on the weight and thickness of the urethral epithelium of castrated female rats. METHODS: Forty castrated adult female Wistar-Hannover rats were randomly divided into two groups: Group I (n=20) in which the animals received only the vehicle (propylene glycol) and Group II (n=20) in which the rats received tamoxifen 250microg/day by gavage. After 30 days of treatment, all animals were sacrificed and the urethra was immediately removed for weighing. Next, the urethra was divided into the proximal and distal segments, which were fixed in 10% formaldehyde and submitted to routine histological techniques for morphometric study. The data were analyzed using the weighted minimum mean-square error method and Student's t-test for two independent samples (p<0.05). RESULTS: There was a significant increase in the mean weight of the urethra in the rats of Group II compared to the control group, 32.0+/-2.0mg and 22.0+/-1.6mg, respectively (p<0.001). The mean thickness of the distal urethral epithelium of the animals treated with tamoxifen was significantly greater than that of the control group, 42.8+/-2.0microm and 36.6+/-1.5microm, respectively (p<0.001). There was no statistically significant difference between the two groups with respect to the epithelial thickness of the proximal urethra (p=0.514). CONCLUSION: Treating castrated adult rats with 250microg/day of tamoxifen for 30 days may increase the weight of the urethra and the thickness of the distal urethral epithelium.

CUSUM chart to monitor autocorrelated counts using Negative Binomial GARMA model.

Cumulative sum control charts have been used for health surveillance due to its efficiency to detect soon small shifts in the monitored series. However, these charts may fail when data are autocorrelated. An alternative procedure is to build a control chart based on the residuals after fitting autoregressive moving average models, but these models usually assume Gaussian distribution for the residuals. In practical health surveillance, count series can be modeled by Poisson or Negative Binomial regression, this last to control overdispersion. To include serial correlations, generalized autoregressive moving average models are proposed. The main contribution of the current article is to measure the impact, in terms of average run length on the performance of cumulative sum charts when the serial correlation is neglected in the regression model. Different statistics based on transformations, the deviance residual, and the likelihood ratio are used to build cumulative sum control charts to monitor counts with time varying means, including trend and seasonal effects. The monitoring of the weekly number of hospital admissions due to respiratory diseases for people aged over 65 years in the city São Paulo-Brazil is considered as an illustration of the current method.

Combined effect of PCV10 and meningococcal C conjugate vaccination on meningitis mortality among children under five years of age in Brazil.

The 10-valent pneumococcal conjugate vaccine (PCV10) was introduced in the Brazilian National Immunization Program in March 2010, scheduled at 2, 4, and 6 months, with a booster at 12-15 months of age. The meningococcal C conjugate vaccine (MCC) was introduced in November 2010, scheduled at 3 and 5 months, with a booster dose at 12-15 months of age and no catch-up for older age groups. In this interrupted time-series analysis study, we used Brazilian mortality data from 2005 to 2015 for children under five years of age (excluding data from the state of Bahia) to assess the combined impact of these vaccines on the overall burden of meningitis mortality among children aged 0-23 months and 2-4 years, as defined using meningitis and meningococcemia specific International Classification of Diseases - tenth revision codes. Secular trends and seasonality were taken into account. We found significant reductions for both age groups relative to those observed for the comparison group of diseases, with immediate effects after the transition period (2010-2011) of 29.2% and 27.5% for children aged 0-23 months and 2-4 years, respectively. These immediate effects were sustained throughout the post-vaccination period (2012-2015). In total, 337 deaths were averted by the combined effect of both vaccines, 238 (95%CI 169-319) for children aged 0-23 months and 99 (95%CI 56-144) for those aged 2-4 years. These results add strong evidence in support of investments in these vaccines by low and middle-income countries.

Genetic polymorphism of calcium-sensing receptor in women with breast cancer.

Breast cancer is a disease of unknown etiology, whose major risk factors are genetic alterations. Polymorphism of the calcium-sensing receptor (CaSR) has been a focus of some recent studies, due to a probable association with breast cancer risk and tumor aggressiveness. A relationship between polymorphic rs17251221 variant of the CaSR gene, and allele G (considered a gain-of-function mutation) and breast cancer risk has been stressed, despite the paucity of studies found in the literature. The present study involved 137 women (69 women with breast cancer-case; and 68 controls without breast cancer) who had 3 ml of peripheral blood drawn for DNA study. Genomic DNA was extracted from leukocytes by genotyping technique with real-time polymerase chain reaction. The AG genotype (rs17251221) was present in 13 women (18.84%) from the case group and in 8 (11.76%) women from the control group (p = 0.3434), while the GG genotype (rs17251221) did not occur in any group. In contrast, no statistically significant difference was observed between the AG genotype of variant rs17251221 in premenopausal case and control women (p = 0.71). There was also no statistically significant difference between postmenopausal case and control patients (p = 0.6851). In the current study, CaSR gene polymorphism of SNP variant rs17251221 did not show any statistically significant association with breast cancer, in both premenopausal and postmenopausal women.

Morphological and morphometric analysis of the adrenal cortex of androgenized female rats.

AIM: We evaluated the thickness of the adrenal cortex zones of female rats androgenized to mimic polycystic ovary syndrome. METHODS: Forty-four female virgin Wistar-Hannover rats were divided into two groups: controls (n = 17) and animals which received testosterone propionate on the 2nd day of life (n = 27). At 90 days of life, after confirmation of persistent estrus, the animals were sacrificed, and the adrenal cortex zones were evaluated. Student's t test and Levene's test were used in the statistical analysis (p < 0.05 considered significant). RESULTS: The adrenal glands of the androgenized rats were more voluminous and had a more intensely vascularized zona reticularis than the control animals. The mean thicknesses of zona glomerulosa and zona reticularis in the androgenized rats were 58.4 and 730.7 mum, respectively, significantly thicker than the values in the control group (45.0 and 328.3 mum, respectively). CONCLUSION: Zona reticularis and zona glomerulosa of the androgenized female rats were significantly thicker than those of the control animals.

CUSUM control charts to monitor series of Negative Binomial count data.

To detect outbreaks of diseases in public health, several control charts have been proposed in the literature. In this context, the usual generalized linear model may be fitted for counts under a Negative Binomial distribution with a logarithm link function and the population size included as offset to model hospitalization rates. Different statistics are used to build CUSUM control charts to monitor daily hospitalizations and their performances are compared in simulation studies. The main contribution of the current paper is to consider different statistics based on transformations and the deviance residual to build control charts to monitor counts with seasonality effects and evaluate all the assumptions of the monitored statistics. The monitoring of daily number of hospital admissions due to respiratory diseases for people aged over 65 years in the city São Paulo-Brazil is considered as an illustration of the current proposal.

Mortality among blood donors seropositive and seronegative for Chagas disease (1996-2000) in São Paulo, Brazil: A death certificate linkage study.

BACKGROUND: Individuals in the indeterminate phase of Chagas disease are considered to have mortality rates similar to those of the overall population. This study compares mortality rates among blood donors seropositive for Chagas disease and negative controls in the city of São Paulo, Brazil. METHODOLOGY/PRINCIPAL FINDINGS: This is a retrospective cohort study of blood donors from 1996 to 2000: 2842 seropositive and 5684 seronegative for Chagas disease. Death status was ascertained by performing probabilistic record linkage (RL) with the Brazil national mortality information system (SIM). RL was assessed in a previous validation study. Cox Regression was used to derive hazard ratios (HR), adjusting for confounders. RL identified 159 deaths among the 2842 seropositive blood donors (5.6%) and 103 deaths among the 5684 seronegative (1.8%). Out of the 159 deaths among seropositive donors, 26 had the 10th International Statistical Classification of Diseases and Related Health Problems (ICD-10) indicating Chagas disease as the underlying cause of death (B57.0/B57.5), 23 had ICD-10 codes (I42.0/I42.2/I47.0/I47.2/I49.0/I50.0/I50.1/ I50.9/I51.7) indicating cardiac abnormalities possibly related to Chagas disease listed as an underlying or associated cause of death, with the others having no mention of Chagas disease in part I of the death certificate. Donors seropositive for Chagas disease had a 2.3 times higher risk of death due to all causes (95% Confidence Interval (95% CI), 1.8-3.0) than seronegative donors. When considering deaths due to Chagas disease or those that had underlying causes of cardiac abnormalities related to Chagas disease, seropositive donors had a risk of death 17.9 (95% CI, 6.3-50.8) times greater than seronegative donors. CONCLUSIONS/SIGNIFICANCE: There is an excess risk of death in donors seropositive blood for Chagas disease compared to seronegative donors. Chagas disease is an under-reported cause of death in the Brazilian mortality database.

Comparative study between ultrasound-guided fine needle aspiration cytology of axillary lymph nodes and sentinel lymph node histopathology in early-stage breast cancer.

The replacement of sentinel lymph node biopsy (SNB) by ultrasound-guided fine-needle aspiration (US-guided FNA) cytology of axillary lymph nodes is controversial, despite the simplicity and reduced cost of the latter. In the present study, US-guided FNA was performed in 27 patients with early-stage breast cancer for comparison with SNB. Data were analyzed by calculation of sample proportions. Tumor subtypes included invasive ductal carcinoma (85%), invasive lobular carcinoma (7%), and tubular and metaplastic carcinoma (4%). FNA had a sensitivity of 45%, specificity of 100%, positive predictive value of 100% and a negative predictive value of 73%. Axillary lymph node cytology obtained by US guided-FNA in patients with breast cancer had a specificity similar to that of sentinel lymph node histopathology in the presence of axillary node metastases. However, when lymph node cytology is negative, it does not exclude the existence of metastatic implants, due to its low sensitivity in comparison to sentinel lymph node histopathology.