Evaluation of leucocytes from sputum samples of pulmonary tuberculosis patients using flow cytometry.

SETTING: Information about the sputum cells of pulmonary tuberculosis (PTB) patients is scarce. The analysis of sputum cells using optical microscopy (OM) is a well-established method, but it has some serious limitations. OBJECTIVE: To establish a new flow cytometry (FC) protocol for the leucocyte evaluation of sputum samples from PTB patients. DESIGN: A new FC protocol using 0.1% dithiothreitol and 0.5% paraformaldehyde was developed to fluidise sputum samples and kill Mycobacterium tuberculosis, respectively, to allow the analysis of sputum samples collected from TB patients. The protocol was validated by comparing it with OM, and the cellularity of 30 sputum samples from patients with PTB was evaluated. RESULTS: The comparison between leucocyte subsets analysed using OM and FC showed agreement. Immunophenotyping of leucocytes from sputum samples showed that neutrophils (95.7%) comprised the largest proportion of sputum cells, followed by monocytes/macrophages (2.6%) and lymphocytes (1.6%). Among the total T-lymphocytes (100%), 12.3% were T-helper cells, 24.1% were cytotoxic T-cells and 62.9% were gamma/delta T; none of the T lymphocytes had the CD4+/CD8+ phenotype. CONCLUSION: FC is a useful method for evaluating the different subtypes of leucocytes present in the sputum samples of PTB patients.

Assessment of physical activity intensity and duration in the paediatric population: evidence to support an a priori hypothesis and sample size in the agreement between subjective and objective methods.

We aimed to provide evidence for an a priori hypothesis and sample size for subjectively assessing physical activity intensity and duration in paediatric population, adopting objective methods as the reference. We searched electronic databases, reference lists and author databases. Correlation coefficients were pooled as an indicator of agreement estimates. We found 183 agreement analyses (94.5% based on correlation coefficients) from 89 studies that met our inclusion criteria. We identified four physical activity parameters addressing intensity and two parameters addressing duration. The physical activity parameters focusing on intensity were measured only by questionnaires, and the best correlation was achieved by moderate-to-vigorous physical activity compared with a heart rate monitor. In addition, total physical activity duration had a stronger correlation with an accelerometer when measured by diaries or logs than when measured by questionnaires. In both cases, the correlation was moderate. Studies with sample sizes between 50 and 99 subjects showed measurements that were stable in both magnitude and interpretation. Our findings suggest that the agreement between subjective and objective methods for assessing physical activity intensity and duration is weak to moderate. Furthermore, sample sizes ranging from 50 to 99 subjects provide stable agreement estimates between methods.

Abdominal obesity and its association with socioeconomic factors among adolescents from different living environments.

BACKGROUND: Socioeconomic status has been associated with obesity in children and adolescents. This association may be dependent according with where adolescents lives. OBJECTIVE: The aim of this study was to evaluate the association between different socioeconomic indicators such as parental education and occupation and socioeconomic status with abdominal obesity in adolescents from two observational studies: the Healthy Lifestyle in Europe by Nutrition in Adolescence cross-sectional study (HELENA-CSS) and the Brazilian Cardiovascular Adolescent Health (BRACAH) study. METHODS: European (n = 3192, aged 12.5-17.5 years, with 53.1% girls from HELENA-CSS) and Brazilian (n = 991, aged 14-18 years, with 54.5% girls from BRACAH study) adolescents from two cross-sectional studies were included in this analysis. Complete data on waist circumference (WC), height, socioeconomic status indicators and several confounders were collected. Socioeconomic indicators were measured using a self-reported questionnaire in order to assess the family social status of the adolescents. Multilevel linear regression models were used to examine associations, and results were adjusted for potential confounders. RESULTS: Adjusted results showed inverse associations between mother's and father's education levels (p < 0.001) and father's occupation level (p < 0.001) with waist-to-height ratio (WHtR) and WC in HELENA-CSS girls. Similarly in European girls, socioeconomic indicators by socioeconomic status and maternal occupation level were associated with WHtR. In HELENA-CSS boys, the same significant association was found between WHtR and WC with maternal occupation level. Moreover, in European boys WHtR was also associated with parental education. In Brazilian adolescents, both indicators of abdominal obesity did not remain associated with the independents variables, after adjustment for potential confounders. CONCLUSIONS: Abdominal obesity was associated with socioeconomic indicators in higher-income countries, but this association was not observed in a lower-middle-income country.

Molecular Cytotoxic Mechanisms of 1-(3,4,5-Trihydroxyphenyl)-dodecylbenzoate in Human Leukemia Cell Lines.

Recent studies have shown that gallic acid and its alkylesters induce apoptosis in different cell lines. Since new compounds with biological activity and less cytotoxicity to normal cells are necessary for cancer therapy, the aim of this study was to evaluate the cytotoxic effect of 1-(3,4,5-trihydroxyphenyl)-dodecylbenzoate on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells. The cell viability was determined by MTT method. The apoptosis induction was assessed by bromide and acridine orange staining and by Annexin V-FITC Apoptosis Detection kit. The cell cycle analysis was carried out by flow cytometry using propidium iodide. Cytometric analysis was also performed to evaluate the expression of the following proteins: AIF, p53, Bcl-2 and Bax. The mitochondrial potential was also assessed by flow cytometry using MitoView633 kit. The results showed that the compound significantly reduced the cell viability of K562 and Jurkat cells in a concentration and time dependent manner (IC50 of 30 µM). The compound induced cell cycle arrest in G0/G1phase and significantly increased the proportion of cells in the sub-G0/G1phase. Apoptosis was confirmed by the sight of morphological characteristics of apoptosis and by phosphatidylserine externalization (73.47±5.71% of cells expressing annexin). The results also showed that the compound promotes a modification in Bax:Bcl-2 ratio and increases p53 expression. Thus, it is possible to conclude that 1-(3,4,5-trihydroxyphenyl)-dodecylbenzoate induces apoptosis by inhibiting the antiapoptotic protein Bcl-2 and by increasing the release of AIF, Bax and p53. In addition, it blocks the cell cycle at G0/G1, stopping cell proliferation. So far, the results suggest that this compound may have a potential therapeutic effect against leukemia cells.

Inflammation profile in overweight/obese adolescents in Europe: an analysis in relation to iron status.

BACKGROUND/OBJECTIVES: The objectives of this study were to investigate the relationship between inflammatory parameters (CRP, c-reactive protein; AGP, α1-acid glycoprotein), iron status indicators (SF, serum ferritin; sTfR, soluble transferrin receptor) and body mass index (BMI) z-score, fat-free mass (FFM) and fat mass (FM) in European adolescents. Differences in intake for some nutrients (total iron, haem and non-haem iron, vitamin C, calcium, proteins) were assessed according to BMI categories, and the association of nutrient intakes with BMI z-score, FM and FFM was evaluated. METHODS: A total of 876 adolescents participating in the Healthy Lifestyle in Europe by Nutrition in Adolescence-Cross Sectional Study were included in the study sample. RESULTS: Mean CRP values (standard error; s.e.) were significantly higher in overweight/obese adolescents (1.7±0.3 and 1.4±0.3 mg/l in boys and girls, respectively) than in thin/normal-weight adolescents (1.1±0.2 and 1.0±0.1 mg/l in boys and girls, respectively) (P<0.05). For boys, mean SF values (s.e.) were significantly higher in overweight/obese adolescents (46.9±2.7 µg/l) than in thin/normal-weight adolescents (35.7±1.7 µg/l) (P<0.001), whereas median sTfR values did not differ among BMI categories for both boys and girls. Multilevel regression analyses showed that BMI z-score and FM were significantly related to CRP and AGP (P<0.05). Dietary variables did not differ significantly among BMI categories, except for the intake of vegetable proteins, which, for boys, was higher in thin/normal-weight adolescents than in overweight/obese adolescents (P<0.05). CONCLUSIONS: The adiposity of the European adolescents was sufficient to cause chronic inflammation but not sufficient to impair iron status and cause iron deficiency.

Prevalence of cardiovascular risk factors among Latin American adolescents: a multilevel analysis.

High blood pressure (HBP) and obesity is a well-established major risk factor for stroke and coronary heart disease. However, the literatures are scarce about these informations in adolescents from low-and-middle income countries. This school-based survey was carried out among students from Maringá (Brazil) and Buenos Aires (Argentina) selected random sampling. We studied 991 Brazilian adolescents (54.5% girls) in the age range of 14-18 years. In Argentina, we studied 933 adolescents (45.9% female) in the age range of 11-17 years. The outcomes of this study are general obesity, abdominal obesity and HBP. The associated factors analysed were gender, age and health behaviours. The prevalence of obesity was 5.8% in Brazil and 2.8% in Argentina, the prevalence of abdominal obesity was 32.7% in Brazil and 11.1% in Argentina, the prevalence of HBP was 14.9% in Brazil and 13.5% in Argentina. The multilevel analysis showed that older adolescents (>14 years old) have a little likelihood of being overweight, whereas male adolescents are more likely to be obese and have HBP. The abdominal obesity in both indicators were not associated with the independent variables. The prevalence of cardiovascular risk factors is high in Latin American adolescents independent of each country, and was associated with male gender.

Sleep time and cardiovascular risk factors in adolescents: the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) study.

OBJECTIVE: We aimed to examine the association between adolescents' sleep time and a cardiometabolic risk score. A second aim was to examine associations between sleep time and individual cardiometabolic risk factors. METHODS: Adolescents (N=699; ages, 12.5-17.5 years) participating in the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) study were examined. Sleep time was reported by a questionnaire. Physical activity (PA) was assessed by accelerometry (n=497). Cardiometabolic risk factors measurements included insulin resistance, blood pressure, adiposity markers, cardiorespiratory fitness, and blood lipids. A cardiovascular disease risk score was computed. Associations were examined by a multilevel regression analysis (linear for individual risk factors and Poisson for the clustered risk score). RESULTS: For school days no association was found between sleep time and cardiometabolic risk factors. At weekend days, the prevalence ratio (PR) of having a clustered risk score increased by 15% for each additional hour of sleep controlling for age, sex, and socioeconomic status (SES); however, the prevalence disappeared when adjusting for PA. CONCLUSIONS: In European adolescents sleep time is not associated with cardiometabolic risk factors when important confounders are considered. Future research about sleep cardiovascular risk factors should register other sleep dimensions (sleep patterns or disturbances) to provide a better insight in this scientific field.

Sedentary behaviour and clustered metabolic risk in adolescents: the HELENA study.

BACKGROUND AND AIMS: Although sedentary behaviours are linked with mortality for cardiovascular reasons, it is not clear whether they are negatively related with cardio-metabolic risk factors. The aim was to examine the association between time engaged in television (TV) viewing or playing with videogames and a clustered cardio-metabolic risk in adolescents. METHODS AND RESULTS: Sedentary behaviours and physical activity were assessed in 769 adolescents (376 boys, aged 12.5-17.5 years) from the HELENA-CSS study. We measured systolic blood pressure, HOMA index, triglycerides, TC/HDL-c, VO2max and the sum of four skinfolds, and a clustered metabolic risk index was computed. A multilevel regression model (by Poisson) was performed to calculate the prevalence ratio of having a clustered metabolic risk. In boys, playing >4 h/day with videogames (weekend) and moderate to vigorous PA (MVPA) was associated with cardio-metabolic risk after adjustment for age, maternal education and MVPA. In contrast, TV viewing was not associated with the presence of cardio-metabolic risk. CONCLUSION: In boys, playing with videogames may impair cardio-metabolic health during the adolescence. Adolescents should be encouraged to increase their participation in physical activity of at least moderate intensity to obtain a more favourable risk factor profile.

Prevalence of abdominal obesity in adolescents: a systematic review.

The objectives of this study were to (i) review extant literature on the prevalence of abdominal obesity (AO) in adolescents of both sex (10-19 years old); (ii) analyse the cut-off points used for the diagnosis of AO and (iii) compare its prevalence between developed and developing countries. The search was carried out using online databases (MEDLINE, Web of Science, EMBASE, SPORTDiscus, SCIELO and BioMed Central), references cited by retrieved articles and by contact with the authors, considering articles published from the establishment of the databanks until 19 October 2009. Only original articles and those using waist circumference in the diagnosis were considered. Twenty-nine studies met the inclusion criteria. Fourteen of these studies were performed in developed countries. The prevalence of AO varied from 3.8% to 51.7% in adolescents from developing countries. The range of results was smaller among developed countries; with values from 8.7% to 33.2%. Eighteen different cut-off points were used. It was concluded the AO prevalence is high among adolescents, but is not clear what sex has a higher proportion and it is greater in adolescents from developing countries; however, there is no consensus in the literature about the criteria to be used.

The relationship between rating of perceived exertion and muscle activity during exhaustive constant-load cycling.

The aims of this study were to verify the relationship between rating of perceived exertion (RPE) and electromyography (EMG) increases during exhaustive constant-load cycling bouts and, to compare and to correlate the power outputs corresponding to perceived exertion threshold (PET) and neuromuscular fatigue threshold (NFT). 11 men completed 3-4 different exhaustive constant-load cycling bouts on a cycle ergometer, being RPE and EMG measured throughout the bouts. The linear regression of the RPEslope and EMGslope against the power output identified the PET and NFT intensity, respectively. There was a significant relationship between RPEslope and EMGslope (R(2)=0.69; P<0.01). However, the linearity of RPEslope (R(2)=0.93±0.07) was significantly higher (P<0.001) than EMGslope (R(2)=0.63±0.25). In addition, the RPEslope and EMGslope were related to time to exhaustion (r=-0.59 and r=-0.60; P<0.001). There was no significant difference (P=0.42) between PET (201.5±27.9W) and NFT (210.3±22.6W) and they were significantly correlated (r=0.78; P=0.005). Therefore, the RPE and EMG increases during exhaustive constant-load cycling bouts are related and, PET and NFT intensities are similar and closely associated.

Electromyography response of the iliocostalis lumborum muscle during flexion, extension and rotation movements of the trunk in orthostatic and seated position.

The action of the column extensor muscles has been studied through in the electromyography in various corporal postures. We verified the electromyography responses of the iliocostalis lumborum muscle during various movements and positions. 12 individuals males, sedentary, between 36 and 52 years old (average 46.16), weighing an average of 79.66 kilos, and with an average height of 173.0 cm, all working as drivers were studied. Electromyography with six channels and surface electrodes was used. The electrodes were placed on the right side of the trunk in the direction of the third and fifth lumbar vertebrae ICL2 and ICL1 respectively). In orthostatic position, the flexion movement presented RMS 31.18 (+/- 5.73) in ICL1 and 58.10 (+/- 14.81) in ICL2; in extension the RMS 32.46 (+/- 9.17) in ICL1 and 55.31 (+/- 16.70) in ICL2; in homolateral rotation the RMS 30.05 (+/- 10.60) in ICL1 and 53.29 (+/- 19.70) in ICL2; in heterolateral rotation the RMS 24.76 (+/- 5.99) in ICL1 and 44.79 (+/- 15.26) in ICL2. In seated in a chair without a back, the flexion presented RMS 22.42 (+/- 2.89) in ICL1 and 43.39 (+/- 5.68) in ICL2; in extension the RMS 22.47 (+/- 1.95) in ICL1 and 41.28 (+/- 6.20) in ICL2; in homolateral rotation the RMS 24.10 (+/- 3.97) in ICL1 and 47.94 (+/- 5.80) in ICL2; in heterolateral rotation, the RMS 22.59 (+/- 1.95) in ICL1 and 43.15 (+/- 5.71) in ICL2. In the position seated on the ground, the flexion presented RMS 22.83 (+/- 3.00) in ICL1 and 50.99 (+/- 11.19) in ICL2; in extension the RMS 22.39 (+/- 3.22) in ICL1 and 46.44 (+/- 8.19) in ICL2. The results are expressed in RMS, indicating the participation of the iliocostalis lumborum muscle during the movements undertaken. The place identified as ICL2 presented action potential of greater amplitude than the place identified as ICL1 in all movements and positions.

[Activation of the cardiac angiotensin-converting enzyme after myocardial infarction and its role in ventricular remodeling]. / Ativação da enzima conversora de angiotensina no coração após infarto do miocárdio e suas repercussões no remodelamento ventricular.

PURPOSE: To determine angiotensin converting enzyme (ACE) activity in the heart of infarcted rats and to investigate the effects of captopril and losartan on the post-infarction remodeling process. METHODS: Myocardial infarction (MI) was produced in Wistar rats by ligature of the left coronary artery. Control rats (Con) underwent a sham surgery. MI and Con rats remained untreated or were treated with captopril (30 mg/kg/day) or losartan (15 mg/kg/day) for 30 days. ACE activity was determined in right (RV) and left ventricular (LV) muscles and in the scar tissue. The effects of captopril therapy was also investigated in the hydroxiproline (OH-Pro) and protein in RV and LV. RESULTS: ACE activity increased 25% in the RV and 70% in the remaining LV muscle. The highest ACE activity was found in the scar tissue, where it was 4.5 times the value of the LV muscle (420 +/- 68 vs 94 +/- 8 nmoles/g/min; P < 0.01). An increase of the end-diastolic pressure and of the muscle mass was found in the RV and LV of MI rats. Captopril and losartan treatments were equally efficient to attenuate these parameters in both ventricles. Captopril also reduced the total OH-Pro content in the RV and LV muscles. The Prot concentration was significantly reduced in the myocardium of MI rats, an effect enhanced by captopril therapy. CONCLUSION: The AII concentration in the blood draining from the scar to the surrounding muscle is probably high. It is likely that this elevated local generation of AII contributes to hypertrophy and to collagen deposition. The effects of ACE inhibitors on remodeling are likely to depend on the reduction of the locally generated AII.