Adropin and Irisin in Patients with Cardiac Cachexia / Adropina e Irisina em Pacientes com Caquexia Cardíaca

Abstract Background: Cardiac cachexia is an important predictive factor of the reduction in survival of patients with heart failure with reduced ejection fraction. Objectives: The aims of the present study were to evaluate adropin and irisin levels in cachectic and non-cachectic subjects and the relationships between the levels of these proteins and clinical and laboratory parameters in patients with HFrEF. Methods: The clinical records of patients who were admitted to the cardiology outpatient clinic for heart failure with reduced ejection fraction were screened. Cachectic patients were identified and assigned to the study group (n = 44, mean age, 65.4 ± 11.2 y; 61.4% men). Heart failure with reduced ejection fraction patients without weight loss were enrolled as the control group (n = 42, mean age, 61.0 ± 16.5 y; 64.3% men). The serum adropin and irisin levels of all patients were measured. A p-value < 0.05 was considered significant. Results: Serum adropin and irisin levels were significantly higher in the cachexia group than in the controls (Adropin (ng/L); 286.1 (231.3-404.0) vs 213.7 (203.1-251.3); p < 0.001, Irisin (µg/mL); 2.6 (2.2-4.4) vs 2.1 (1.8-2.4); p = 0.001). Serum adropin and irisin levels were positively correlated with brain natriuretic peptide (BNP) levels and New York Heart Association (NYHA) class and negatively correlated with body mass index (BMI) and serum albumin levels (all p values: < 0.001). In a multivariate analysis, adropin was the only independent predictor of cachexia in the heart failure with reduced ejection fraction patients (OR: 1.021; 95% CI: 1.004−1.038; p = 0.017). Conclusions: The results suggest that adropin and irisin may be novel markers of cardiac cachexia in heart failure with reduced ejection fraction patients. Adropin and irisin are related with the severity of heart failure.

Resumo Fundamento: A caquexia cardíaca é um importante preditor de redução de sobrevida em pacientes com insuficiência cardíaca com fração de ejeção reduzida (ICFER). O objetivo deste estudo foi avaliar os níveis de adropina e irisina em pacientes com ICFER caquéticos e não caquéticos, assim como a relação entre os níveis dessas proteínas e os parâmetros clínicos e laboratoriais nesses pacientes. Objetivos: Os objetivos do presente estudo foram avaliar os níveis de adropina e irisina em indivíduos caquéticos e não caquéticos e as relações entre os níveis dessas proteínas e os parâmetros clínicos e laboratoriais em pacientes com ICFEN. Métodos: Os prontuários de pacientes atendidos no ambulatório de cardiologia para ICFER foram triados. Aqueles com ICFER caquéticos foram identificados e constituíram o grupo de estudo (n = 44; idade média, 65,4 ± 11,2 anos; 61,4% de homens). Aqueles com ICFER e sem perda de peso foram arrolados como grupo controle (n = 42; idade média, 61,0 ± 16,5 anos; 64,3% de homens). Os níveis séricos de adropina e irisina de todos os pacientes foram medidos. Considerou-se significativo um p-valor < 0,05. Resultados: Os níveis séricos de adropina e irisina foram significativamente mais altos nos pacientes caquéticos do que nos controles [adropina (ng/l): 286,1 (231,3-404,0) vs 213,7 (203,1-251,3); p < 0,001; irisina (µg/ml): 2,6 (2,2-4,4) vs 2,1 (1,8-2,4); p = 0,001]. Os níveis séricos de adropina e irisina correlacionaram-se positivamente com os níveis de peptídeo natriurético cerebral (BNP) e a classe funcional da New York Heart Association (NYHA), e negativamente com o índice de massa corporal (IMC) e os níveis séricos de albumina (todos os p-valores: < 0,001). Na análise multivariada, a adropina foi o único preditor independente de caquexia nos pacientes com ICFER (OR: 1,021; IC 95%: 1,004−1,038; p = 0,017). Conclusões: Os resultados sugerem que a adropina e a irisina possam ser novos marcadores de caquexia cardíaca em pacientes com ICFER. Adropina e irisina estão relacionadas com a gravidade da insuficiência cardíaca.

Biodisponibilidade de peptídeos do feijão caupi (Vigna unguiculata L. Walp) e o metabolismo do colesterol / Bioavailability of cowpea peptides (Vigna unguiculata L. Walp) and the cholesterol metabolism

Introdução: Doenças cardiovasculares constituem importante causa de morte em todo mundo e a hipercolesterolemia está diretamente relacionada a elas. A dieta desempenha papel importante neste processo e alguns alimentos como o feijão caupi (Vigna unguiculata L. Walp), especialmente sua proteína, tem sido apontado com potencial capacidade de redução do colesterol plasmático. Os efeitos hipocolesterolêmicos já observados indicaram o uso da proteína do feijão caupi, ou dos seus peptídeos, como ingrediente funcional de alimentos para a promoção da saúde e a redução do risco de doenças. Entretanto, as consequências da digestão gastrointestinal na absorção destes peptídeos são claramente complexas tornando essenciais estudos in vitro e in vivo para avaliar a sua bioacessibilidade e sua resistência à degradação gastrointestinal, além da disponibilidade e real eficácia destes peptídeos. Objetivo: Analisar a biodisponibilidade de peptídeos e avaliar parâmetros ligados ao metabolismo do colesterol em modelos animais após ingestão de isolado proteico de feijão caupi. Métodos: A farinha de feijão caupi foi desengordurada e sua proteína isolada. O isolado proteico foi submetido a métodos de hidrólise in vitro, para verificação das frações peptídicas formadas e inferência sobre a capacidade de ligação à albumina. Dois experimentos in vivo foram conduzidos. No primeiro, o isolado proteico do feijão caupi foi administrado a ratos e a concentração dos peptídeos monitorada no sangue, por 2 horas. O experimento in vivo 2 consistiu na alimentação de hamsters com dietas normo (N) - e hipercolesterolêmicas por 21 dias, contendo a proteína do feijão caupi como única proteína da ração (I), comparada ao controle de caseína (H). Neste experimento foram analisados no plasma: colesterol total (CT) e frações (LDLc, VLDLc e HDLc), triglicerídeos (TG) e peptídeos; nas fezes: colesterol total (CF) e ácidos biliares (AB); no fígado: colesterol (CH) e lipídeos totais (LH), HMGCR (atividade enzimática e expressão) e expressão de SREBP2, LDLR, ABCA1, ABCG1, ABCG5, ABCG8, LXRa e AMPK. Resultados: Os peptídeos identificados a partir da hidrólise proteica do feijão caupi, ou a partir do plasma dos animais estudados não evidenciaram similaridades entre os experimentos ou corresponderam a sequências previamente identificadas para o feijão caupi a partir de banco de dados. CT, VLDLc, HDLc, TG, CH dos hamsters foram maiores nos grupos H e I quando comparado ao N; LDLc foi maior para I comparado aos demais; LH foi maior em H comparado a N, sendo que I não diferiu dos demais; CF foi maior para I comparado a N, sendo que H não diferiu dos demais. A expressão de ABCA1 foi maior para I em relação aos demais; LXRa foi maior para I em relação a H, mas N não diferiu dos demais; SREBP2 foi menor em H em comparação aos demais; HMGCR foi mais expressa em N em comparação aos demais, ao passo que a atividade desta enzima foi maior em I quando comparado a N, sendo que H não diferiu dos demais. Não houve diferença entre os grupos quanto a AB ou expressão de ABCG8 ou AMPK. Não foram obtidos resultados de expressão para LDLR, ABCG1 e ABCG5. Conclusão: Apesar de pesquisas anteriores a este trabalho terem evidenciado a capacidade do isolado proteico do feijão caupi em inibir a atividade da HMGCR, inibir a solubilização micelar ou melhorar o perfil de lipídeos plasmáticos, no trabalho atual esta matéria prima não mostrou atuação positiva quanto ao metabolismo do colesterol de hamsters nas condições experimentais utilizadas. Os fragmentos indicados como peptídeos obtidos a partir da hidrólise proteica do feijão caupi, ou do plasma dos animais estudados não corresponderam a peptídeos com comprovada, ou até mesmo, com indicação de bioatividade

Introduction: Cardiovascular diseases are important cause of death worldwide and hypercholesterolemia is directly related to them. Diet plays an important role in this process and some foods such as cowpea (Vigna unguiculata L. Walp), especially its protein, have been shown to have a potential for reducing plasma cholesterol. The hypocholesterolemic effects already observed indicated the use of cowpea protein, or its peptides, as a functional food ingredient for health promotion and reduction of disease risk. However, the consequences of gastrointestinal digestion on the absorption of these peptides are clearly complex, making in vitro and in vivo studies essential to assess their bioaccessibility and resistance to gastrointestinal degradation, as well as the availability and actual efficacy of these peptides. Objectives: To analyze the bioavailability of peptides and evaluate parameters related to cholesterol metabolism in animal models after ingestion of protein isolate of cowpea. Methods: Cowpea flour was defatted and its protein isolated. The protein isolate was subjected to in vitro hydrolysis methods to verify the formed peptide fractions and inference about albumin binding ability. Two in vivo experiments were conducted. In the first, the cowpea protein isolate was administered to rats and the concentration of the peptides monitored in the blood for 2 hours. The in vivo experiment 2 consisted of feeding hamsters with normal (N) - and hypercholesterolemic diets for 21 days, containing the cowpea protein as the sole dietary protein (I), compared to casein control (H). In this experiment were analyzed in the plasma: total cholesterol (TC) and fractions (LDLc, VLDLc and HDLc), triglycerides (TG) and peptides; In feces: total cholesterol (CF) and bile acids (AB); In the liver: cholesterol (CH) and total lipids (LH), HMGCR (enzymatic activity and expression) and expression of SREBP2, LDLR, ABCA1, ABCG1, ABCG5, ABCG8, LXRa and AMPK. XX. Results: The peptides identified from the protein hydrolysis of cowpea or from the plasma of the animals studied did not show similarities among the experiments or correspond to sequences previously identified for the cowpea from the database. CT, VLDLc, HDLc, TG, CH of hamsters were higher in groups H and I when compared to N; LDLc was higher for I compared to the others; LH was higher in H compared to N, and I did not differ from the others; CF was higher for I compared to N, and H did not differ from the others. The expression of ABCA1 was higher for I than the others; LXRa was higher for I than H, but N did not differ from the others; SREBP2 was lower in H compared to the others; HMGCR was more expressed in N compared to the others, whereas the activity of this enzyme was higher in I when compared to N, and H did not differ from the others. There was no difference between groups regarding AB or expression of ABCG8 or AMPK. No expression results were obtained for LDLR, ABCG1 and ABCG5. Conclusion: Although previous research to this work evidenced the ability of the cowpea protein isolate to inhibit HMGCR activity, inhibit micellar solubilization or improve the plasma lipid profile, in the current work this raw material did not show a positive cholesterol metabolism of hamsters under the experimental conditions used. The fragments indicated as peptides obtained from the protein hydrolysis of cowpea beans, or from the plasma of the animals studied did not correspond to peptides with proven, or even, with indication of bioactivity

Experimental osteonecrosis: development of a model in rodents administered alendronate

Abstract The main objective of this study was to cause bisphosphonate-related osteonecrosis of the jaws to develop in a rodent model. Adult male Holtzman rats were assigned to one of two experimental groups to receive alendronate (AL; 1 mg/kg/week; n = 6) or saline solution (CTL; n = 6). After 60 days of drug therapy, all animals were subjected to first lower molar extraction, and 28 days later, animals were euthanized. All rats treated with alendronate developed osteonecrosis, presenting as ulcers and necrotic bone, associated with a significant infection process, especially at the inter-alveolar septum area and crestal regions. The degree of vascularization, the levels of C-telopeptide cross-linked collagen type I and bone-specific alkaline phosphatase, as well as the bone volume were significantly reduced in these animals. Furthermore, on radiographic analysis, animals treated with alendronate presented evident sclerosis of the lamina dura of the lower first molar alveolar socket associated with decreased radiographic density in this area. These findings indicate that the protocol developed in the present study opens new perspectives and could be a good starting model for future property design.

Proteomic Analysis of Rat Brain Stem with DAI by MALDI-TOF-MS / 法医学杂志

OBJECTIVE@#To establish a diagnostic model for diffuse axonal injury (DAI) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). To screen the proteins or peptides associated with DAI for providing the biomarkers with theoretic foundation.@*METHODS@#Fifteen male Sprague-Dawley rats were randomly divided into DAI group (n = 10) and control group (n = 5). The protein or peptide expression profiles of rat brain stem were detected by MALDI-TOF-MS. ClinProTools 2.2 software was used to find specific peaks, and a diagnostic model was established by the genetic algorithm.@*RESULTS@#There were significant differences in 61 peaks of DAI group (P < 0.05), 9 peaks were down-regulated and 52 up-regulated. The diagnostic model was established based on 5 different peaks. The specificity and sensitivity of cross validation was 96.14% and 95.98%; while the specificity and sensitivity of blind validation showed was 73.33% and 70.00%, respectively.@*CONCLUSION@#A specific and sensitive diagnostic model of DAI can be established by MALDI-TOF-MS to provide a potential value for determining DAI in forensic practice.

Bone health in cerebral palsy and introduction of a novel therapy / A saúde do osso na paralisia cerebral e a introdução de uma nova terapia

ABSTRACT Objective To assess the bone health status of children with cerebral palsy and the therapeutic effect of denosumab in a subgroup of children with cerebral palsy and decreased bone mass. Methods Children with cerebral palsy were evaluated according to their motor disability score (classification system gross motor functions III to V), bone density and bone turnover markers. Dual X-ray energy absorption was used to measure the lumbar spine, and total body, except the head. Thereafter a group of children with cerebral palsy and osteoporosis was treated with denosumab, a fully human monoclonal antibody. Bone turnover markers were measured before and three months after treatment. Results Reduction in bone mineral density was observed, particularly in children with greater impairment evaluated by the motor score. Decreased bone turnover markers were found in a selected group of children three months after exposure to denosumab. Conclusion Bone loss was present in children with significant impairment of motor function, as well as decreased serum levels of bone resorption markers with new forms.

RESUMO Objetivo Avaliar o estado de saúde dos ossos em crianças com paralisia cerebral e o efeito terapêutico do denosumabe em um subgrupo de crianças com paralisia cerebral e redução da massa óssea. Métodos Crianças com paralisia cerebral foram avaliadas de acordo com seu escore de incapacidade motora (sistema de classificação para funções motoras grossas, de III a V), e marcadores de turnover ósseo. Dual de absorção de energia de raios X foi utilizado para medir a coluna lombar e total do corpo menos cabeça. Posteriormente, um grupo de crianças com paralisia cerebral e osteoporose foi tratado com denosumabe, um anticorpo monoclonal totalmente humano. Marcadores de remodelação óssea foram medidos antes e três meses após o tratamento. Resultados Houve uma redução da densidade óssea, particularmente em crianças com maior comprometimento do escore motor; os marcadores de remodelação óssea diminuíram em um grupo selecionado de crianças três meses depois de terem sido expostas ao denosumabe. Conclusão A perda óssea esteve presente em crianças com importante comprometimento das funções motoras, além da redução nos níveis séricos de marcadores de reabsorção óssea com novos tratamentos.

Biomarcadores óseos y su relación con la densidad mineral ósea en adultas jóvenes y adolescentes en su primer año posparto / Bone biomarkers and its relation with bone mineral density in adults and adolescents during the first year postpartum

La información sobre biomarcadores óseos en adolescentes y adultas durante el periodo posparto es incierta, por lo que el objetivo de este artículo fue analizar el patrón de biomarcadores óseos en adolescentes y adultas a 15, 90, 180 y 365 días posparto (dpp) y su asociación con la densidad mineral ósea (DMO) y lactancia materna. Se realizó un estudio de cohorte en 32 madres adolescentes ≤17 años y 41 adultas de 18 a 29 años de edad en el primer año posparto. Se realizaron medidas antropométricas, DMO y biomarcadores óseos y así como datos del tipo y la duración de lactancia. Como resultados se encontró asociación entre la concentración basal de N-telopéptidos ≤24 μg/L y mayor aumento de DMO. Las adolescentes tuvieron mayor concentración de N-telopéptidos (p≤0.004) y menor concentración de osteocalcina (5±3 vs13±4, p <0.001) que las adultas. La lactancia no afectó el cambio de DMO (p>0.050), ni de biomarcadores óseos. La osteocalcina se asoció con el cambio en DMO (p<0.040). La prolactina fue mayor entre las que practicaron lactancia materna exclusiva (p<0.001). A menor edad menores concentraciones de osteocalcina (p<0.001) y mayores concentraciones de N-telopéptidos (p<0.001). Se concluyó que a menor concentración de N-telopéptidos y mayor de osteocalcina hubo un mayor aumento de DMO, lo cual implica menor aumento de ésta en el grupo de adolescentes. La lactancia no afectó la DMO.

The objective of this study was to describe the trend of bone biomarkers in adults and adolescents women at 15, 90, 180 and 365 postpartum days (ppd) and its relation with bone mineral density (BMD). It was a prospective cohort of 32 teenager’s ≤17 and 41 women from 18 to 29 years old. We evaluated diet, anthropometry, BMD, bone biomarkers and hormonal profile. In all, the concentration of N-telopeptide was higher at 15 days postpartum decreasing during first year postpartum, but adolescents had the highest concentration. The lowest N-telopeptide concentration was associated with highest increasing of the BMD. Osteocalcin concentration was lower in adolescents than in adults women (5 ± 3 vs 13 ± 4 ng/mL, p<0.001) during first year postpartum. Exclusive breastfeeding did not affect the BMD (p>0.050) or bone biomarkers. Osteocalcin concentration was positively associated with bone BMD (p<0.040), breastfeeding did not affect osteocalcin concentrations. Prolactin was higher among women who breastfed exclusively (p<0.001). Age and breastfeeding inversely correlated with bone biomarkers (p<0.001) N-telopeptide and PTHi respectively. We concluded that a lower N-telopeptide concentration and a higher osteocalcin concentration were associated with a higher increasing of BMD, so then, adolescents showed the lowest recovery of the BMD. Breastfeeding does not affect the BMD.

Early effects of tumor necrosis factor inhibition on bone homeostasis after soluble tumor necrosis factor receptor use

BACKGROUND/AIMS: Our aim was to assess whether short-term treatment with soluble tumor necrosis factor (TNF) receptor affects circulating markers of bone metabolism in rheumatoid arthritis (RA) patients. METHODS: Thirty-three active RA patients, treated with oral disease-modifying antirheumatic drugs (DMARDs) and glucocorticoids for > 6 months, were administered etanercept for 12 weeks. Serum levels of bone metabolism markers were compared among patients treated with DMARDs at baseline and after etanercept treatment, normal controls and naive RA patients not previously treated with DMARDs (both age- and gender-matched). RESULTS: Bone-specific alkaline phosphatase (BSALP) and serum c-telopeptide (CTX)-1 levels were lower in RA patients treated with DMARDs than in DMARD-naive RA patients. After 12 weeks of etanercept treatment, serum CTX-1 and sclerostin levels increased. In patients whose DAS28 improved, the sclerostin level increased from 1.67 +/- 2.12 pg/mL at baseline to 2.51 +/- 3.03 pg/mL, which was statistically significant (p = 0.021). Increases in sclerostin levels after etanercept treatment were positively correlated with those of serum CTX-1 (r = 0.775), as were those of BSALP (r = 0.755). CONCLUSIONS: RA patients treated with DMARDs showed depressed bone metabolism compared to naive RA patients. Increases in serum CTX-1 and sclerostin levels after short-term etanercept treatment suggest reconstitution of bone metabolism homeostasis.

Proteomic profiling of nephrotic syndrome in serum using magnetic bead based sample fractionation & MALDI-TOF MS.

Background & objectives: At present, the diagnosis of nephrotic syndrome (NS) requires a renal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate serum petidome in NS patients. Methods: The fasting blood samples from 49 patients diagnosed with NS by renal biopsy, including 17 mesangial proliferative glomerulonephritis (MsPGN), 12 minimal change nephrotic syndrome (MCNS), 10 focal segmental glomerulosclerosis (FSGS) and 10 membranous nephropathy (MN), were collected and screened to describe their variability of the serum peptidome. The results in NS group were compared with those in 10 control healthy individuals. Specimens were purified with magnetic beads-based weak cation exchange chromatography and analyzed in a MALDI-TOF MS. Results: The results showed 43, 61, 45 and 19 differential peptide peaks in MsPGN, MCNS, MN and FSGS groups, respectively. A Genetic Algorithm was used to set up the classification models. Cross validation of healthy controls from MsPGN, MCNS, MN and FSGS was 96.18, 100, 98.53 and 94.12 per cent, respectively. The recognition capabilities were 100 per cent. Interpretation & conclusions: Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may give an early idea of the pathology of nephrotic syndrome.

Serum Elastin-Derived Peptides and Anti-Elastin Antibody in Patients with Systemic Sclerosis

The elastin metabolism in systemic sclerosis (SSc) has been known to be abnormal. The authors investigated relationship between the clinical manifestations of systemic sclerosis (SSc) and serum levels of soluble elastin-derived peptide (S-EDP) and anti-elastin antibodies. Serum samples were obtained from 79 patients with SSc and 79 age- and sex-matched healthy controls. Concentrations of serum S-EDP and anti-elastin antibodies were measured by ELISA. The serum concentrations of S-EDP in SSc patients were significantly higher than in healthy controls (median, 144.44 ng/mL vs 79.59 ng/mL, P < 0.001). Serum EDP concentrations were found to be correlated with disease duration in SSc (P = 0.002) and particularly in diffuse cutaneous SSc (P = 0.005). Levels of anti-elastin antibodies were found to be more elevated in SSc patients than in healthy controls (median, 0.222 U vs 0.191 U, P = 0.049), more increased in diffuse cutaneous SSc than limited cutaneous SSc (median, 0.368 U vs 0.204 U, P = 0.031). In addition, levels of anti-elastin antibodies were also found to be negatively associated with presence of anti-centromere antibody (P = 0.023). The S-EDP levels were not found to be correlated with levels of anti-elastin antibodies. The increased S-EDP and anti-elastin antibody levels and association with clinical and laboratory characteristics may reflect the abnormal metabolism in SSc.

Current Recommendations for Laboratory Testing and Use of Bone Turnover Markers in Management of Osteoporosis

Osteoporosis is a major health problem worldwide, and is projected to increase exponentially due to the aging of the population. The absolute fracture risk in individual subjects is calculated by the use of algorithms which include bone mineral density (BMD), age, gender, history of prior fracture and other risk factors. This review describes the laboratory investigations into osteoporosis which include serum calcium, phosphate, creatinine, alkaline phosphatase and 25-hydroxyvitamin D and, additionally in men, testosterone. Parathyroid hormone (PTH) is measured in patients with abnormal serum calcium to determine its cause. Other laboratory investigations such as thyroid function testing, screening for multiple myeloma, and screening for Cushing's syndrome, are performed if indicated. Measurement of bone turnover markers (BTMs) is currently not included in algorithms for fracture risk calculations due to the lack of data. However, BTMs may be useful for monitoring osteoporosis treatment. Further studies of the reference BTMs serum carboxy terminal telopeptide of collagen type I (s-CTX) and serum procollagen type I N-terminal propeptide (s-PINP) in fracture risk prediction and in monitoring various treatments for osteoporosis may help expedite their inclusion in routine clinical practice.

Impacto da maturação esquelética em biomarcadores do metabolismo ósseo e na densidade mineral óssea em adolescentes brasileiros saudáveis / Impact of skeletal maturation on bone metabolism biomarkers and bone mineral density in healthy Brazilian male adolescents

OBJETIVO: Avaliar o comportamento de biomarcadores de formação e reabsorção óssea em adolescentes brasileiros em função da sua maturação biológica. MÉTODOS: Oitenta e sete voluntários foram divididos em grupos segundo a idade óssea (IO): 10-12 anos (n = 25), 13-15 anos (n = 36) e 16-18 anos (n = 26). Foram analisados peso (kg), estatura (m), índice de massa corporal (kg/m2), ingestão de cálcio de 3 dias (mg/dia), avaliação dos eventos pubertários pelos critérios de Tanner, níveis dos biomarcadores [osteocalcina (OC) (ng/mL), fosfatase alcalina óssea (FAO) (U/L), telopeptídeo carboxiterminal sérico (S-CTx) (ng/mL)] e sua correlação com a densidade mineral óssea (DMO) (g/cm2) por atenuação de raios X de dupla energia da coluna lombar, do fêmur proximal e de corpo total. RESULTADOS: Os biomarcadores mostraram comportamento semelhante, apresentando medianas elevadas dos 13 aos 15 anos (FAO = 154,71 U/L, OC = 43,0 ng/mL, S-CTx = 2,09 ng/mL; p < 0,01) e no estágio puberal G4. As medianas decresceram com o avançar da IO e da maturação sexual. Os níveis dos biomarcadores mostraram paralelismo com pico de velocidade em estatura, e, curiosamente, os biomarcadores de formação indicaram correlação negativa com a DMO, isto é, valores de DMO elevados se correlacionaram com valores baixos dos biomarcadores. CONCLUSÕES: Este é o primeiro estudo em adolescentes brasileiros com critérios de inclusão e exclusão rígidos e cuidadosos a avaliar a correlação entre marcadores ósseos e DMO frente a indicadores da maturação biológica. Os resultados ajudam a compreender o turnover ósseo e o monitoramento do metabolismo ósseo.

OBJECTIVE: To evaluate the behavior of biomarkers of bone formation and resorption in healthy male Brazilian adolescents according to their biological maturation. METHODS: Eighty-seven volunteers were divided into age groups according to bone age (BA): 10-12 years (n = 25), 13-15 years (n = 36), and 16-18 years (n = 26). Weight (kg), height (m), body mass index (kg/m2), calcium intake from 3 days assessed by 24-h food recall (mg/day), pubertal event evaluation by Tanner criteria, and serum biomarker levels (osteocalcin [OC] [ng/mL], bone alkaline phosphatase [BAP] [U/L], and serum carboxyterminal telopeptide [S-CTx] [ng/mL]) were recorded and correlated to bone mineral density (BMD) (g/cm2) measured by dual energy X-ray absorptiometry of the lumbar spine, proximal femur, and whole body. RESULTS: Biomarkers showed similar behaviors, presenting higher median values in the 13-15 year group (BAP = 154.71 U/L, OC = 43.0 ng/mL, S-CTx = 2.09 ng/mL; p < 0.01) and when adolescents were in the pubertal stage G4. Median biomarker values decreased with advancing BA and sexual maturation. Biomarker values showed parallelism with peak height velocity, and, interestingly, bone formation biomarkers indicated significant negative correlation with BMD in the different evaluated locations, i.e., higher BMD values correlated with lower bone biomarker values. CONCLUSIONS: This is the first study of healthy Brazilian adolescents with rigid and careful inclusion and exclusion criteria to assess the correlation of bone markers and BMD with biological maturation indicators. Our results can help understand bone turnover and monitor bone metabolism.

The effects of discontinuing long term alendronate therapy in a clinical practice setting / Efeitos da retirada do uso prolongado de alendronato na prática clínica

OBJECTIVE: To assess bone turnover markers (BTM) and bone mineral density (BMD) after discontinuation of alendronate treatment used for five or more years. SUBJECTS AND METHODS: 40 patients (pt) with post-menopausal osteoporosis treated with alendronate (10 mg/d) for at least five years (Group 1, G1) had their medication discontinued. Group 2 (G2): 25 pt treated with alendronate for at least one year. Group 3 (G3): 23 treatment-naïve osteoporotic pt. BMD was evaluated in G1 and G2 at baseline and after 12 months. Collagen type I cross-linked C-telopeptide (CTX) and procollagen type 1 N-terminal propeptide (P1NP) levels were measured in all pt at baseline, and in G1 and G2 every three months for 12 months. Data were analyzed using ANOVA on ranks and Mann-Whitney tests. RESULTS: Mean BMD values in G1 and G2 did not differ during follow-up. However, 16 pt (45.7 percent) in G1 and one (5.2 percent) in G2 lost BMD (P < 0.001). BTM at baseline was not different between G1 and G2, and both were lower than G3. A significant increase in BTM levels was detected in G1 pt after three months, but not in G2. CONCLUSION: Observed BMD loss and BTM rise after alendronate withdrawal imply that bone turnover was not over suppressed, and alendronate discontinuation may not be safe.

OBJETIVO: Avaliar a evolução dos marcadores de metabolismo ósseo (MMO) e da densidade mineral óssea (DMO) após cinco anos de uso de alendronato em mulheres osteoporóticas na pós-menopausa. SUJEITOS E MÉTODOS: 40 pacientes (pct) osteoporóticas, na pós-menopausa, em uso de alendronato (10 mg/dia) por pelo menos 5 anos (Grupo 1 − G1) tiveram o uso do bisfosfonato suspenso. O grupo 2 (G2): 25 mulheres na pós-menopausa, em uso de alendronato (10 mg/dia) há pelo menos 1 ano. Grupo 3 (G3): 23 pct osteoporóticas, controles ainda sem tratamento. G1 e G2 submeteram-se à avaliação da DMO por DXA (basal e após 12 meses de seguimento). Todas as pct colheram amostras basais de CTX e P1NP, e G1 e G2 submeteram-se a coletas trimestrais de CTX e P1NP durante 1 ano. Resultados foram analisados por ANOVA on ranks e Mann-Whitney. RESULTADOS: Níveis médios de DMO não variaram em G1 ou G2 durante o estudo; no entanto, 16 pct (45,7 por cento) no G1 e 1 pct (5,2 por cento) no G2 apresentaram redução clinicamente significativa de DMO (P < 0,001). Níveis basais de CTX e P1NP não diferiram entre G1 e G2, com ambos inferiores aos níveis de G3. Em G1, observou-se elevação significativa de CTX e P1NP após 3 meses. Os níveis de CTX e P1NP em G2 permaneceram estáveis durante todo o seguimento. CONCLUSÃO: Não parece haver supressão excessiva do metabolismo ósseo na prática clínica. A suspensão temporária do alendronato após seu uso prolongado pode não ser segura.

Normocalcemic primary hyperparathyroidism in clinical practice: an indolent condition or a silent threat? / Hiperparatireoidismo primário normocalcêmico na prática clínica: condição indolente ou ameaça silenciosa?

OBJECTIVE: To describe the characteristics of normocalcemic primary hyperparathyroidism (NPHPT) in patients seen for osteoporosis evaluation. PATIENTS AND METHODS: We examined the records of 156 women who came to the hospital to be screened for osteoporosis. Measurements of total calcium, PTH, 25-hydroxy vitamin D, and β-C-telopeptide were recorded. Bone mineral density and T-scores were evaluated by densitometry of the lumbar spine, femoral neck and distal one-third of the radius. The latter was only measured in patients with primary hyperparathyroidism. Nephrolithiasis and bone fractures were documented by a review of the medical records. RESULTS: We identified 14 patients with NPHPT, accounting for 8.9 percent of the population studied. In the medical records, the occurrence of kidney stones was reported in 28.6 percent of the patients with NPHPT, in contrast with only 0.7 percent of the noncarriers. Regarding the presence of general fractures, 21.4 percent of the patients with NPHPT were affected versus 16.2 percent of noncarriers. CONCLUSION: Data from our study suggest that NPHPT has a diverse phenotypic presentation, implying that this may not be an "indolent" disease.

OBJETIVO: Avaliar as características do hiperparatireoidismo primário normocalcêmico (HPTPN) em pacientes atendidos para avaliação de osteoporose. PACIENTES E MÉTODOS: Foi realizada análise de um banco de dados de 156 mulheres que procuraram atendimento para avaliação de osteoporose. Todas apresentavam dosagem de cálcio sérico, PTH, 25-hidroxi-vitamina D e C-telopeptídeo. A densidade mineral óssea e escore-T foram avaliados por meio de densitometria óssea de coluna lombar, colo do fêmur e rádio distal, este último apenas em pacientes com hiperparatireoidismo renal primário. Nefrolitíase e fraturas ósseas foram documentadas pela revisão dos prontuários. RESULTADOS: Foram identificadas 14 pacientes com HPTPN, correspondendo a 8,9 por cento da população estudada. Nos registros médicos, o relato da existência de litíase renal ocorreu em 28,6 por cento dos portadores de HPTN em contraste com apenas 0,7 por cento nas mulheres não portadoras, com um p < 0,001. CONCLUSÃO: Os dados do estudo sugerem que HPTPN tem uma apresentação fenotípica variada, podendo não ser uma patologia "indolente".

Short-term bone marker responses to teriparatide and strontium ranelate in patients with osteoporosis previously treated with bisphosphonates / Resposta dos marcadores de curto prazo a teriparatida e ranelato de estrôncio em pacientes com osteoporose previamente tratados com bisfosfonatos

OBJECTIVE: To evaluate the responses of serum β-CTX and osteocalcin in patients who were undergoing treatment with teriparatide or strontium ranelate (SR). SUBJECTS AND METHODS: We analyzed 14 patients (12 women and 2 men; mean age of 71 years) taking teriparatide, and 13 female patients (mean age of 70 years) taking SR; all the patients having previously been on bisphosphonates. Serum β-CTX and osteocalcin levels were determined before and after the first and third months of teriparatide treatment and up to the fourth month of treatment with SR. RESULTS: We observed an initial significant increase in osteocalcin levels during the first month (165 percent, p = 0.01) followed by a peak of β-CTX (180 percent, p = 0.02) after the third month of treatment with teriparatide. An increase in these markers was also observed with SR: 49 percent in osteocalcin (p = 0.002) and 80 percent in β-CTX (p = 0.008). CONCLUSION: SR had a predominantly short-term bone-forming effect in postmenopausal women with osteoporosis previously treated with bisphosphonates in a lesser degree than with teriparatide.

OBJETIVO: Avaliar as respostas do β-CTX e osteocalcina séricos em pacientes que foram submetidas a tratamento com teriparatida ou ranelato de estrôncio (RE). SUJEITOS E MÉTODOS: Analisaram-se 14 pacientes (12 mulheres e 2 homens; idade média 71 anos) tomando teriparatida, e 13 mulheres (idade média 70 anos) tomando RE; todos os pacientes haviam previamente tomado bisfosfonatos. Níveis séricos de β-CTX e osteocalcina foram determinados antes e após o primeiro e terceiro meses de tratamento com teriparatida e no quarto mês de tratamento com RE. RESULTADOS: Observou-se um aumento inicial significativo nos níveis de osteocalcina no primeiro mês (165 por cento, p = 0,01), seguido por um pico do β-CTX (180 por cento, p = 0,02) após o terceiro mês de tratamento com teriparatida. Aumento nesses marcadores também foi observado com RE: 49 por cento na osteocalcina (p = 0,002) e 80 por cento no β-CTX (p = 0,008). CONCLUSÃO: RE teve um efeito predominantemente na formação óssea a curto prazo em mulheres na pós-menopausa com osteoporose tratadas previamente com bisfosfonatos em menor grau que a teriparatida.

C telopeptides of type I collagen inpostmenopausal women: an experience in a Tunisian clinical laboratory

The purpose of study was to evaluate the interest of C-telopeptides of type I collagen [CTX] in the diagnosis of osteoporosis in postmenopausal women and to define its cut-off value. A transverse descriptive study enrolled postmenopausal women: 139 osteoporotic [G1] and 39 non osteoporotic [G2]. The 2 groups were defined by bone density measurement. The followmg markers were measured: serum alkaline phosphatase [ALP] bone alkaline phosphatase [bone ALP], serum C-terminal telopeptide of type I collagen [CTX]. Statistical analyses were performed using SPSS 10, 5. The corresponding estimation of sensitivity and specificity of CTX have been presented as receiver Operating Curve [ROC]. There was no difference in the measurement of ALP and bone ALP in the 2 groups but CTX was statistically higher in G1 compared to G2 [p<0.001]. The percentage of osteoporotic women [G1] with CTX values>0.500 ng/ml was higher than that of non osteoporotic women [G2]. We have established a ROC curve to find the cut-off value of CTX that enables the distinction between osteoporotic women with high level of bone remodelling, and non osteoporotic women. The cut-off value of CTX 0.55 pg/mi was the best; it associated best sensitivity and specificity. The total increase and significance for CTX was greater in the group of osteoporotic women and appeared therefore to be a good bone turnover marker in the diagnosis of osteoporosis in comparison with ALP and bone ALP. The cut-off value of CTX 0.55 pg/mi may improve the sensitivity and specificity of prediction of future fractures

Acute Effects of Intravenous Administration of Pamidronate in Patients with Osteoporosis

We investigated acute effects of intermittent large dose bisphophonate therapy in osteoporotic patients. Peripheral blood mononuclear cells were incubated with alendronate (100 micrometer) for 18 hr, in vitro and cytokine expressions were measured by real-time RT-PCR. Pamidronate 30 mg was administered on 26 osteoporotic patients; and acute phase reactants, inflammatory cytokines and bone biomarkers were measured. The in vitro study showed significant increase in mRNA expression of IL-6, TNF-alpha and IFN-gamma. A notable rise in serum C-reactive protein (CRP) was observed over 3 days after pamidronate infusion (P=0.026). Serum levels of TNF-alpha, IL-6 and IFN-gamma were also significantly increased (P=0.009, 0.014, 0.035, respectively) and the increase in IL-6 levels were strongly correlated with CRP levels (P=0.04). Serum calcium and c-telopeptide levels rapidly decreased after the treatment (P=0.02, <0.001, respectively). This study showed that mRNA expression of inflammatory cytokines at peripheral blood mononuclear cells (PBMC) level were observed within 18 hr and marked elevation of inflammatory cytokines and acute phase reactants were demonstrated after pamidronate infusion at the dose for osteoporosis. Our studies confirmed that intermittent large dose aminobisphosphonate causes acute inflammation.

Serum cross-linked n-telopeptides of type 1 collagen (NTx) in patients with solid tumors / Dosagem sérica do N-telopeptídeo do colágeno tipo I (NTx) em pacientes com tumores sólidos

CONTEXT AND OBJECTIVE: Cross-linked N-telopeptides of type I collagen (NTx) increase in concentration in situations in which bone resorption is increased, such as osteoporosis and bone metastasis (BM). We aimed to evaluate the serum concentrations of NTx in a sample of patients with several types of solid tumors. DESIGN AND SETTING: Cross-sectional analytical study with a control group in a tertiary public hospital. METHODS: We performed the quantitative enzyme-linked immunosorbent assay (ELISA) on serum NTx levels in 19 subjects without a history of cancer and 62 patients with various solid tumors who had been referred for a bone scan. Three experienced analysts read all bone scans. RESULTS: The serum NTx levels in patients with cancer and BM, with cancer but without BM and without cancer were 46.77 ± 2.58, 32.85 ± 2.05 and 22.32 ± 2.90 respectively (P < 0.0001). We did not find any significant correlations of serum NTx with age, gender, history of bone pain, tumor type and bone alkaline phosphatase levels. We found a significant correlation between serum NTx and alkaline phosphatase levels (R² = 0.08; P = 0.022). CONCLUSIONS: Serum NTx levels are significantly higher in patients with solid tumors and bone metastases than they are in patients without bone metastases and in normal controls.

CONTEXTO E OBJETIVO: Os N-telopeptídeos do colágeno tipo-I (NTx) elevam-se quando a reabsorção óssea está aumentada, devido a condições como osteoporose e metástase óssea. Sendo assim, temos por objetivo avaliar os níveis séricos de NTx em uma população heterogênea de pacientes com tumores sólidos. TIPO DE ESTUDO E LOCAL: Estudo transversal analítico com grupo controle, realizado em hospital público terciário. MÉTODOS: 19 pacientes sem história de câncer e 62 pacientes com tumores sólidos de vários tipos, encaminhados para estadiamento e investigação dos sintomas esqueléticos, foram avaliados pela técnica de ELISA (Enzyme Linked Immuno Sorbent Assay) quantitativa para a dosagem de NTx. Três especialistas leram todas as imagens ósseas. RESULTADOS: O nível de NTx encontrado em pacientes com câncer e metástase óssea, sem metástase óssea e sem diagnóstico de câncer foi 46,77 ± 2,58, 32,85 ± 2,05 e 22,32 ± 2,90, respectivamente (P < 0,0001). Não encontramos correlação entre o NTx, idade, sexo, história de dor óssea, tipo de tumor e níveis de fosfatase alcalina óssea. Encontramos correlação significativa entre os níveis de NTx e de Fosfatase Alcalina (r² = 0,08, P = 0,022). CONCLUSÃO: O NTx sérico é significativamente mais elevado em pacientes com tumores sólidos e metástases ósseas quando comparado com pacientes sem metástases ósseas e controles normais.

Evaluation of polypeptide YY and ghrelin in complicated type 2 diabetic patients

The gut peptides polypeptide YY [PYY], [a potent satiety agent] and ghrelin [a potent hunger signal] are suggested to play a role in obesity. Obesity is commonly associated or complicated with hypertension [HT] and type 2 diabetes [T2D]. Thus, the current study aimed to: [i] estimate the fasting plasma levels of PYY and ghrelin in lean versus overweight subjects as well as overweight HT and T2D subjects, [ii] assess if PYY and ghrelin are affected by the glycemic state; and [iii] intercorrelate the levels of PYY and ghrehn considering insulin sensitivity, blood pressure values and lipid profile in HT and/or T2D subjects. Twelve lean healthy male subjects [group I] and fifty eight overweight, age and sex matched subjects [group II] were included in the present study. Group II [overweight group] was further sub classified into: [i] group IIa: normoglycemic normotensive subjects [n=14]; [ii] group IIb: T2D normotensive patients [n=18]; [iii] group IIc: normoglycemic HT patients [n=14]. Fasting plasma lipid profile, glucose [FG], insulin [Fl]. PYY, ghrelin and blood glycated hemoglobin A1C [HbA1C] were estimated. Insulin sensitivity was evaluated according to the homeostatic model assessment [HOMA] index. In the present study significantly lower mean plasma levels of both ghrelin and PYY were observed in all overweight groups versus the lean control group. The hypertensive and T2D groups, also, showed lower PYY and ghrelin levels compared to the over-weight normotensive normoglycemic group. Furthermore, in hypertension T2D group both present, both PYY and ghrelin levels showed further decrease. Ghrelin correlated positively with high density lipoprotein cholesterol, HDL-c [r=0.43, p<0.01]. Both ghrelin and PYY correlated negatively with BMI, FG, Fl, HbA1c, HOMA index, low density lipoprotein cholesterol [LDL-c] and mean arterial blood pressure [r=0.52, r=0.62, r=-0.73, r=-0.71, r=-0.76, r=-0.42, and r=-0.5. p<0.01 respectively for ghrelin: and r=-0.51, r=-0.61, r=-0.62, r=-0.39, and r=-0.48, p<0.05 respectively for PYY]. In controls, PYY and ghrelin were negatively correlated [r=-0.76, p<0.001]. However, in all groups of patients studied, they were positively correlated [r=0.64, p<0.001], Multiple regression analysis revealed that low ghrelin and PYY concentration were independently correlated to BMI [p=0.002, and p=0.009 respectively]. Low ghrelin was, also, independently correlated to FI [i.e., hyperinsulinemia] [p=0.04]. In the diabetic groups both PYY and ghrelun levels were lower in patients with poor glycemtc control versus controlled diabetics, as assessed byHbA1C. Thus, from the current study it could be concluded that low PYY and ghrelin levels may play a role in the pathogenesis of obesity, hypertension and T2D. Combination of a ghrelin antagonist [a hunger signal antagonist] and PYY [a satiety signal] is potentially and attractive therapeutic strategy for treatment of obesity and its complications

Evaluación de anticuerpos anti-péptido cíclico citrulinado y anti-vimetina mutada en pacientes con artitris reumatoide / Evaluation of ciclic citrulinated anti-peptide antibodies and anti-vimentin muted in patients with rheumatoid arthritis

La Artritis Reumatoide (AR) es una enfermedad inflamatoria crónica y multisistémica de etiología desconocida, que afecta principalmente las articulaciones periféricas. Su diagnóstico desde el laboratorio se ha limitado al empleo del Factor Reumoatoide (FR) como único marcador considerado por le Colegio Americano de Reumatología (CAR) entre sus criterios de detección para la enfermedad, conociendo su relativa sensibilidad y baja especificidad, surgió el propósito de hallar un marcador seológico con sensibilidad y especificidad suficiente capaz de contribuir al diagnóstico de la AR. Para dar cumplimiento con el objetivo general, se llevó a cabo la evaluación de los ensayos de anticuerpos anti-péptidos cíclicos citrulinados de segunda generación a través de dos metodologías (anti-CCP2, ELISA el Inmunofluorometría-enzimática) y anti-vimentina mutada citrulinada (anti-MCV, ELISA) y su correlación posterior con el FR y anticuerpos anti-nucleares (AAN), por lo cual, se seleccionaron 59 pacientes con AR y 29 pacientes con otras enfermedades autoinmunes. observándose que el ensayo anti-MCV (ELISA) presentó mayor sensibilidad diagnóstica, en tanto que el anti-CCP2 (ELISA) reflejó mayor especificidad, sin embargo, se obtuvo mejor correlación y concordancia entre los ensayos anti CCP2 (Inmunofluorometría-enzimática) y anti-MCV (ELISA). Así mismo se obtuvo moderada concordancia entre el FR y el anti-CCP2 (ELISA) y, entre los AAN y el anti-CCP2 (ELISA). Concluyendo que el anti-MCV (ELISA) presentó un mejor comportamiento para el diagnóstico de la enfermedad, sin dejar en consideración que el anti-CCP2 a través de sus dos metodologías también puede ser utilizado por el Reumatológo en la evaluación del paciente con AR.

The rheumatoid arthritis (RA) is a chronic multisystemic inlfammatory disease of unknown etiology that principally effects the peripheral articulations. It's diagnose from the laboratory has been limited to the emplyment of the rheumatoid factor (RF) as the only marker considerate by the American College of Rheumatology (ACR) between there criteria for the disease detection. Knowing its relative sensibility and low specificity, we proposed to find a serologic marker wih adequate sensibility and specificity to contribute in the diagnosis of RA. To perform the general objective, we evaluated the assays of cyclic citruliated anti-peptide antibodies of second generation by two methodologies (anti-CCP2, ELISA and Enzymatic Immunofuorometry) and muted anti-vimetin citrulinated (anti-MCV, ELISA) and it's correlation with the RF and anti-nuclear antibodies (ANA). There were selected 59 patients with RA and 29 patients with other autoimmune disease. We observed that the anti-MCV (ELISA) assay presented mayor diagnostic sensibility, while the anti-CCP2 (ELISA) reflected mayor specificity. But with he (anti-CCP2, ELISA and Enzimatic Inmunofluoremetry) and anti-MCV (ELISA) we obtained better correlations and concodance. We also obtained moderate concordante between FR and anti-CCP2, (Elisa), and between ANA and anti-CCP2 (ELISA). Concluding that the anti-MCV (ELISA) presented a better behavior for the diagnosis of the disease, without considerating that the anti-CCP2 by it's both methodologies can be utilized by the Rheumatologist for the evaluation of the patient with RA.