Perfil clínico-epidemiológico de pacientes com hanseníase atendidos em hospital universitário no Rio de Janeiro entre 2008 e 2017

Modelo do estudo: Estudo observacional, longitudinal, retrospectivo e descritivo de uma coorte de pacientes. Objetivo: Descrever o perfil clínico e epidemiológico dos pacientes com diagnóstico de hanseníase, atendidos em um centro de referência no estado do Rio de Janeiro. Metodologia: Foram incluídos todos os pacientes com diagnóstico de hanseníase atendidos no Hospital Universitário Gaffrée e Guinle entre os anos de 2008 a 2017. Os dados foram coletados através da revisão de prontuários e analisados através de estatística descritiva, utilizando-se o software SPSS 22.0 database . Resultados: Um total de 112 pacientes foi incluído no estudo, sendo que 54,5% eram do sexo masculino. A média de idade foi de 49,9 anos, 62,4% eram moradores do município do Rio de Janeiro e 69,9% apresentaram formas clínicas multibacilares da hanseníase. Foi possível avaliar o grau de incapacidade inicial de 36 (32,1%) dos pacientes incluídos. Desses, 25% apresentaram grau de incapacidade inicial II. Durante o tratamento poliquimioterápico, 21 (18,8%) pacientes apresentaram algum efeito colateral, sendo o mais comum a anemia relacionada à dapsona. Cinquenta e nove (52,7%) pacientes apresentaram algum tipo de estado reacional da hanseníase e 10 (8,9%) apresentaram complicações sistêmicas graves durante o acompanhamento. Conclusões: O presente estudo evidenciou um perfil clínico-epidemiológico diferente do descrito na literatura, com uma grande porcentagem de pacientes com grau de incapacidade inicial II, uma alta frequência de estados reacionais e de complicações advindas do curso clínico da doença ou dos tratamentos implementados.Introduction: This was an observational, longitudinal, retrospective and descriptive study of a cohort of patients. Objectives: To describe the clinical and epidemiological profile of the patients diagnosed with leprosy admitted to a reference medical center in the State of Rio de Janeiro. Methods: All leprosy patients admitted to the dermatology clinic from the Hospital Universitário Gaffrée and Guinle between 2008 and 2017 were included. Pertinent patients’ data were collected through medical charts review. Data were then analyzed through descriptive statistics using SPSS 22.0  software.  Results: a total of 112 patients were included in the study. Among them, 54.5% were male, the median age was 49.9 years old, 62.4% were residents in the city of Rio de Janeiro and 69.9% presented multibacillary leprosy clinical forms. We evaluated the initial disability grade of 36 from the 112 included patients,  and out of them,  25% had grade II disability. During multidrug therapy, 21 (18.8%) patients presented an adverse effect, which is the most common anemia related to Dapsone. Fifty-nine (52.7%) patients presented leprosy reaction and 10 (8.9%) patients presented severe systemic complications during the follow-up. Conclusions: In the present study, it was observed a distinctive clinical and epidemiological leprosy profile. There was a high frequency of grade II initial disability, leprosy reaction and severe clinical complications due to the progression of the disease or as adverse effects of leprosy treatment

CARACTERÍSTICAS HIDROGEOMORFOMÉTRICAS DA MICROBACIA DO RIO SANTA CRUZ, AMAZÔNIA OCIDENTAL, BRASIL

The conservation of natural resources is essential for sustainable development in agricultural properties, and can be achieved with planning based on detailed information on landscape characteristics. The objective of this work is to carry out the hydrogeomorphometric characterization of the Santa Cruz river microbasin, Western Amazon, Brazil. Data was acquired through equations and geotechnologies. The microbasin has an area of 500.72 km2, perimeter of 115.57 km, form factor of 0.23, compactness coefficient of 1.44, circularity index of 0.47, altitudes between 189 and 484 m, 94% from flat to gently undulating reliefs, drainage network of 491.77 km, dendritic drainage pattern, 5th order river hierarchy, drainage density of 0.98 km km-2, 0.82 springs km-2, maintenance coefficient of 1,018.18 m2 m-1, sinuosity index of 36.36% and concentration time of 13.89 h. Therefore, the microbasin has an elongated shape, low susceptibility to flooding, low susceptibility to water erosion, high potential for agricultural mechanization, complex drainage system, high condition for fish habitation, low densities (hydrographic, drainage and springs), good efficiency maintenance, wandering main channel and high concentration time. The information can be used to form public policies aimed at sustainable development in the region.La conservación de los recursos naturales es esencial para el desarrollo sostenible en las propiedades agrícolas, y puede obtenerse con una planificación basada en información detallada de las características del paisaje. El trabajo tiene como objetivo realizar la caracterización hidrogeomorfométrica de la cuenca del río Santa Cruz, Amazonas Occidental, Brasil. Los datos se adquirieron mediante ecuaciones y geotecnias. La cuenca tiene una superficie de 500,72 km2, un perímetro de 115,57 km, un factor de forma de 0,23, un coeficiente de compacidad de 1,44, un índice de circularidad de 0,47, altitudes entre 189 y 484 m, un 94% de relieve plano a suavemente ondulado, una red de drenaje de 491,77 km, un patrón de drenaje dendrítico, una jerarquía fluvial de 5º orden, una densidad de drenaje de 0,98 km-2, 0,82 manantiales km-2, un coeficiente de mantenimiento de 1. 018,18 m2 m-1, índice de sinuosidad de 36,36% y tiempo de concentración de 13,89 h. Por lo tanto, la cuenca tiene una forma alargada, baja susceptibilidad a las inundaciones, baja susceptibilidad a la erosión del agua, alto potencial de mecanización agrícola, sistema de drenaje complejo, alta condición para la habitabilidad de los peces, bajas densidades (hidrograma, drenaje y manantiales), buena eficiencia de mantenimiento, canal principal ramificado y alto tiempo de concentración. La información puede utilizarse para la formación de políticas públicas dirigidas al desarrollo sostenible de la región.  A conservação dos recursos naturais é essencial para o desenvolvimento sustentável nas propriedades agropecuárias, e pode ser obtida com um planejamento alicerçado em informações detalhadas das características da paisagem. O trabalho tem como objetivo realizar a caracterização hidrogeomorfométrica da microbacia do rio Santa Cruz, Amazônia Ocidental, Brasil. Os dados foram adquiridos por meio de equações e geotecnologias. A microbacia tem área de 500,72 km2, perímetro de 115,57 km, fator de forma de 0,23, coeficiente de compacidade de 1,44, índice de circularidade de 0,47, altitudes entre 189 e 484 m, 94% de relevos planos a suave ondulados, rede de drenagem de 491,77 km, padrão de drenagem dendrítico, hierarquia fluvial de 5ª ordem, densidade de drenagem de 0,98 km km-2, 0,82 nascentes km-2, coeficiente de manutenção de 1.018,18 m2 m-1, índice de sinuosidade de 36,36% e tempo de concentração de 13,89 h. Portanto, a microbacia tem formato alongado, baixa suscetibilidade a enchentes, baixa suscetibilidade a erosão hídrica, elevado potencial para mecanização agrícola, sistema de drenagem complexo, alta condição para a habitação de peixes, baixas densidades (hidrográfica, drenagem e nascentes), boa eficiência de manutenção, canal principal divagante e elevado tempo de concentração. As informações podem ser utilizadas para a formação de políticas públicas direcionadas para o desenvolvimento sustentável da região.A conservação dos recursos naturais é essencial para o desenvolvimento sustentável nas propriedades agropecuárias, e pode ser obtida com um planejamento alicerçado em informações detalhadas das características da paisagem. O trabalho tem como objetivo realizar a caracterização hidrogeomorfométrica da microbacia do rio Santa Cruz, Amazônia Ocidental, Brasil. Os dados foram adquiridos por meio de equações e geotecnologias. A microbacia tem área de 500,72 km2, perímetro de 115,57 km, fator de forma de 0,23, coeficiente de compacidade de 1,44, índice de circularidade de 0,47, altitudes entre 189 e 484 m, 94% de relevos planos a suave ondulados, rede de drenagem de 491,77 km, padrão de drenagem dendrítico, hierarquia fluvial de 5ª ordem, densidade de drenagem de 0,98 km km-2, 0,82 nascentes km-2, coeficiente de manutenção de 1.018,18 m2 m-1, índice de sinuosidade de 36,36% e tempo de concentração de 13,89 h. Portanto, a microbacia tem formato alongado, baixa suscetibilidade a enchentes, baixa suscetibilidade a erosão hídrica, elevado potencial para mecanização agrícola, sistema de drenagem complexo, alta condição para a habitação de peixes, baixas densidades (hidrográfica, drenagem e nascentes), boa eficiência de manutenção, canal principal divagante e elevado tempo de concentração. As informações podem ser utilizadas para a formação de políticas públicas direcionadas para o desenvolvimento sustentável da região

Dietary inflammatory index and inflammatory biomarkers in adolescents from LabMed physical activity study

Background/objectives The dietary inflammatory index (DII) is a tool to measure the diet’s inflammatory potential and has been used with adults to predict low-grade inflammation. The present study aims to assess whether this dietary score predicts low-grade inflammation in adolescents. Subjects/methods The sample comprises 329 adolescents (55.9% girls), aged 12–18 years, from LabMed Physical Activity Study. DII score was calculated based on a food-frequency questionnaire and categorized into tertiles. We collected blood samples to determine the follow inflammatory biomarkers: C-reactive protein (CRP), interleukin-6 (IL-6), complement component 3 (C3), and 4 (C4). In addition we calculated an overall inflammatory biomarker score. Odds ratios (OR) and 95% confidence intervals (95%CI) were computed from binary logistic regression models. Results DII score, comparing first with third tertile, was positively associated with IL-6 in crude model (OR = 1.88, 95% CI:1.09–3.24, ptrend = 0.011) and in fully adjusted (for biological and lifestyle variables) (OR = 3.38, 95%CI:1.24–9.20, ptrend = 0.023). Also, DII score was positively associated with C4, when fully adjusted (OR = 3.12, 95%CI:1.21–8.10, ptrend = 0.016). DII score was negatively associated with C3 in crude model, comparing first with second but not with third tertile, and no significant associations in fully adjusted model were observed, although a trend was found (OR = 1.71, 95% CI:0.63–4.66, ptrend = 0.044). No significant associations were observed between DII score and CRP. However, DII score was positively associated with the overall inflammatory biomarker score, when fully adjusted (OR = 5.61, 95% CI:2.00–15.78, ptrend = 0.002). Conclusions DII score can be useful to assess the diet’s inflammatory potential and its association with low-grade inflammation in adolescents.The authors gratefully acknowledged the participation of all adolescents and their parents, teachers and schools of the LabMed and Physical Activity Study, the cooperation of volunteer’s, the Department of Hygiene and Epidemiology (University of Porto) for the conversion food frequency questionnaire data into nutrients, and the Research Centre in Physical Activity, Health and Leisure (University of Porto) for the sponsoring the LabMed and Physical Activity Study.info:eu-repo/semantics/publishedVersio

High anti-SARS-CoV-2 antibody seroconversion rates before the second wave in Manaus, Brazil, and the protective effect of social behaviour measures: results from the prospective DETECTCoV-19 cohort

Background: The city of Manaus, Brazil, has seen two collapses of the health system due to the COVID-19 pandemic. We report anti-SARS-CoV-2 nucleocapsid IgG antibody seroconversion rates and associated risk factors in Manaus residents before the second wave of the epidemic in Brazil. Methods: A convenience sample of adult (aged ≥18 years) residents of Manaus was recruited through online and university website advertising into the DETECTCoV-19 study cohort. The current analysis of seroconversion included a subgroup of DETECTCoV-19 participants who had at least two serum sample collections separated by at least 4 weeks between Aug 19 and Oct 2, 2020 (visit 1), and Oct 19 and Nov 27, 2020 (visit 2). Those who reported (or had no data on) having a COVID-19 diagnosis before visit 1, and who were positive for anti-SARS-CoV-2 nucleocapsid IgG antibodies at visit 1 were excluded. Using an in-house ELISA, the reactivity index (RI; calculated as the optical density ratio of the sample to the negative control) for serum anti-SARS-CoV-2 nucleocapsid IgG antibodies was measured at both visits. We calculated the incidence of seroconversion (defined as RI values ≤1·5 at visit 1 and ≥1·5 at visit 2, and a ratio >2 between the visit 2 and visit 1 RI values) during the study period, as well as incidence rate ratios (IRRs) through cluster-corrected and adjusted Poisson regression models to analyse associations between seroconversion and variables related to sociodemographic characteristics, health access, comorbidities, COVID-19 exposure, protective behaviours, and symptoms. Findings: 2496 DETECTCoV-19 cohort participants returned for a follow-up visit between Oct 19 and Nov 27, 2020, of whom 204 reported having COVID-19 before the first visit and 24 had no data regarding previous disease status. 559 participants were seropositive for anti-SARS-CoV-2 nucleocapsid IgG antibodies at baseline. Of the remaining 1709 participants who were seronegative at baseline, 71 did not meet the criteria for seroconversion and were excluded from the analyses. Among the remaining 1638 participants who were seronegative at baseline, 214 showed seroconversion at visit 2. The seroconversion incidence was 13·06% (95% CI 11·52–14·79) overall and 6·78% (5·61–8·10) for symptomatic seroconversion, over a median follow-up period of 57 days (IQR 54–61). 48·1% of seroconversion events were estimated to be asymptomatic. The sample had higher proportions of affluent and higher-educated people than those reported for the Manaus city population. In the fully adjusted and corrected model, risk factors for seroconversion before visit 2 were having a COVID-19 case in the household (IRR 1·49 [95% CI 1·21–1·83]), not wearing a mask during contact with a person with COVID-19 (1·25 [1·09–1·45]), relaxation of physical distancing (1·31 [1·05–1·64]), and having flu-like symptoms (1·79 [1·23–2·59]) or a COVID-19 diagnosis (3·57 [2·27–5·63]) between the first and second visits, whereas working remotely was associated with lower incidence (0·74 [0·56–0·97]). Interpretation: An intense infection transmission period preceded the second wave of COVID-19 in Manaus. Several modifiable behaviours increased the risk of seroconversion, including non-compliance with non-pharmaceutical interventions measures such as not wearing a mask during contact, relaxation of protective measures, and non-remote working. Increased testing in high-transmission areas is needed to provide timely information about ongoing transmission and aid appropriate implementation of transmission mitigation measures. Funding: Ministry of Education, Brazil; Fundação de Amparo à Pesquisa do Estado do Amazonas; Pan American Health Organization (PAHO)/WHO.World Health OrganizationRevisión por pare

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

ATLANTIC EPIPHYTES: a data set of vascular and non-vascular epiphyte plants and lichens from the Atlantic Forest

Epiphytes are hyper-diverse and one of the frequently undervalued life forms in plant surveys and biodiversity inventories. Epiphytes of the Atlantic Forest, one of the most endangered ecosystems in the world, have high endemism and radiated recently in the Pliocene. We aimed to (1) compile an extensive Atlantic Forest data set on vascular, non-vascular plants (including hemiepiphytes), and lichen epiphyte species occurrence and abundance; (2) describe the epiphyte distribution in the Atlantic Forest, in order to indicate future sampling efforts. Our work presents the first epiphyte data set with information on abundance and occurrence of epiphyte phorophyte species. All data compiled here come from three main sources provided by the authors: published sources (comprising peer-reviewed articles, books, and theses), unpublished data, and herbarium data. We compiled a data set composed of 2,095 species, from 89,270 holo/hemiepiphyte records, in the Atlantic Forest of Brazil, Argentina, Paraguay, and Uruguay, recorded from 1824 to early 2018. Most of the records were from qualitative data (occurrence only, 88%), well distributed throughout the Atlantic Forest. For quantitative records, the most common sampling method was individual trees (71%), followed by plot sampling (19%), and transect sampling (10%). Angiosperms (81%) were the most frequently registered group, and Bromeliaceae and Orchidaceae were the families with the greatest number of records (27,272 and 21,945, respectively). Ferns and Lycophytes presented fewer records than Angiosperms, and Polypodiaceae were the most recorded family, and more concentrated in the Southern and Southeastern regions. Data on non-vascular plants and lichens were scarce, with a few disjunct records concentrated in the Northeastern region of the Atlantic Forest. For all non-vascular plant records, Lejeuneaceae, a family of liverworts, was the most recorded family. We hope that our effort to organize scattered epiphyte data help advance the knowledge of epiphyte ecology, as well as our understanding of macroecological and biogeographical patterns in the Atlantic Forest. No copyright restrictions are associated with the data set. Please cite this Ecology Data Paper if the data are used in publication and teaching events. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Effect of physical exercise on cellular immune response of patients with Chronic Obstructive Pulmonary Disease (COPD)

Os estágios avançados da DPOC são longos e dolorosos processos onde há o aumento dos sintomas, fazendo com que o paciente entre em um ciclo vicioso de deterioração da capacidade física, dispneia, ansiedade e isolamento social. Deste modo, o exercício vem se mostrando um componente importante na DPOC, auxiliando no tratamento medicamentoso para a redução dos sintomas e melhora da qualidade de vida. Neste contexto, não há muitos relatos na literatura sobre o papel da atividade física no padrão de secreção de citocinas e na resposta proliferativa de pacientes com DPOC. Além disso, não há muita concordância sobre o comprimento do telômero e fenótipo celular quando a comparados tabagistas que não desenvolvem a doença e pacientes com DPOC. O objetivo deste trabalho foi comparar alguns parâmetros imunológicos entre pacientes com DPOC e indivíduos tabagistas sem DPOC, e em pacientes com DPOC antes e após o programa de reabilitação oferecido no Hospital das Clínicas da FMUSP. As coletas de sangue foram realizadas em dois momentos para o grupo DPOC (pré e pós-programa de reabilitação pulmonar), e em um único momento para o grupo tabagista. Estas amostras foram processadas para obtenção de células mononucleares do sangue periférico, onde foram analisados os seguintes parâmetros: proliferação celular e apoptose, fenotipagem de linfócitos, comprimento relativo do telômero e dosagem de citocinas. Verificamos que indivíduos tabagistas possuem menores quantidades de proteína C reativa que pacientes com DPOC, e uma tendência a maior número de linfócitos. Além disso, o comprimento relativo do telômero em tabagistas é maior do que em pacientes com DPOC, especialmente em linfócitos TCD8+, e em menor grau em linfócitos TCD4+. Linfócitos TCD8+ de portadores de DPOC apresentaram maiores porcentagens de células terminalmente diferenciadas, sugerindo exaustão celular destes linfócitos, e menores porcentagens de células de memória central e memória efetora. Pacientes com DPOC apresentam maiores quantidades de citocinas comparados aos tabagistas sem DPOC. Já na comparação pré e pós-reabilitação verificamos menores quantidades de leucócitos, menores pontuações nos questionários de sintomas, e maiores distâncias percorridas no teste de caminhada de 6 minutos. Na avaliação da linfoproliferação, para as células estimuladas com mitógeno (fitohemaglutinina) e antígenos (citomegalovirus e Haemophilus influenza) foi possível verificar melhora na resposta linfoproliferativa dos pacientes no período pós-reabilitação, assim como maiores níveis da citocina imunoreguladora IL-10. Deste modo concluímos que pacientes com DPOC possuem um perfil mais pró-inflamatório e de diferenciação terminal que tabagistas sem a doença e que exercício físico é capaz de modular o ambiente inflamatório melhorando alguns parâmetros da resposta imune celularThe advanced stages of COPD are long and painful with increase of symptoms making the patients enter a vicious cycle of deterioration of physical capacity, dyspnea, anxiety and social isolation. Therefore, the exercise shows an important component of COPD pathogenesis, assisting in pharmacological treatment, reducing symptoms and enhancing life quality. In this context, there are no concise reports in literature about the role of physical activity in the pattern of cytokine secretion and proliferative response in COPD patients. Besides this, there is no consensus in the data comparing smokers with COPD and smokers without COPD. Because of this, we compared some immune parameters of smokers with COPD and smokers without COPD, and evaluated the cellular immune response before and after a rehabilitation program offered at the Hospital das Clínicas FMUSP. Blood collection was performed in two moments in the COPD group (before and after the rehabilitation program), and once in smokers without COPD. After that the samples were processed to peripheral blood mononuclear cells isolation, in which were analyzed the cellular proliferation and apoptosis, the lymphocyte phenotypic characteristics, the telomere length and the cytokines levels in serum and culture supernatants. Smokers without COPD have lower levels of C reactive protein, and a trend to greater percentages of lymphocytes than in smoker with COPD. The telomere length of COPD patients was shorter than that of smokers without COPD, especially in TCD8+ lymphocytes, with a non-significant trend in the TCD4+ lymphocytes. The TCD8+ subpopulation of COPD patients comprised greater percentages of terminally differentiated cells, and lower percentages of central memory and effector memory cells, suggesting a bias to more terminally differentiated (and eventually exhausted) cells. Moreover, the levels of pro inflammatory cytokines were greater in COPD patients. Evaluation of the exercise effect, we found greater quantities of leucocytes, lower scores in the symptoms questionnaires and longer distances in the six minute walk test after the rehabilitation program. Besides this, the proliferative response to the mitogen phytohemaglutinin, and the antigens from cytomegalovirus and nontypeable Haemophilus influenza were all improved after the exercise program with greater levels of secretion of the anti-inflammatory cytokine IL-10. In conclusion, COPD patients have a pro inflammatory profile and a bias for more terminally differentiated (and eventually exhausted) cells when compared with smokers without COPD, and that the exercise program is capable of modulating the inflammatory microenvironment enhancing some parameters of the cellular immune respons

Cost-Effective Trap qPCR Approach to Evaluate Telomerase Activity: an Important Tool for Aging, Cancer, and Chronic Disease Research

OBJECTIVES: Telomeres are a terminal ‘‘DNA cap’’ that prevent chromosomal fusion and degradation. However, aging is inherent to life, and so is the loss of terminal sequences. Telomerase is a specialized reverse transcriptase encoded by self-splicing introns that counteract chromosome erosion. Telomerase activity is observed during early embryonic development, but after the blastocyst stage, the expression of telomerase reduces. The consequences of either insufficient or unrestrained telomerase activity underscore the importance of ongoing studies aimed at elucidating the regulation of telomerase activity in humans. In the present study, we aimed to standardize a simplified telomerase repeat-amplification protocol (TRAP) assay to detect telomerase activity in unstimulated and PHA-stimulated mononuclear cells. METHODS and RESULTS: Our optimized qPCR-based can efficiently evaluate telomerase activity. Quantification of protein and DNA between unstimulated and PHA-stimulated peripheral blood mononuclear cells revealed cellular activation and cell-cycle entry. The assay also showed that relative telomerase activity is significantly different between these two conditions, supporting the applicability of the assay. Furthermore, our findings corroborated that telomerase activity decreases with age. CONCLUSIONS: Telomeres and telomerase are implicated in aging and development of chronic diseases and cancer; however, difficulty in accessing commercial kits to investigate these aspects is a critical constraint in health surveillance studies. Our optimized assay was successfully used to differentiate telomerase activity between unstimulated and stimulated cells, clearly showing the reactivation of telomerase upon cell activation. This assay is affordable, reproducible, and can be executed in resource-limited settings