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1.
Aging Male ; 21(3): 200-205, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29327639

RESUMO

Erectile dysfunction is a common disease characterized by endothelial dysfunction. The aetiology of ED is often multifactorial but evidence is being accumulated in favor of the proper function of the vascular endothelium that is essential to achieving and maintaining penile erection. Uric acid itself causes endothelial dysfunction via decreased nitric oxide production. This study aims to evaluate the serum uric acid (SUA) levels in 180 ED patients, diagnosed with the International Index of Erectile Function-5 (IIEF-5) and 30 non-ED control. Serum uric acid was analyzed with a commercially available kit using ModularEVO (Roche, Monza, Italy). Within-assay and between-assay variations were 3.0% and 6.0%, respectively. Out of the ED patients, 85 were classified as arteriogenic (A-ED) and 95 as non-arteriogenic (NA-ED) with penile-echo-color-Doppler. Uric acid levels (median and range in mg/dL) in A-ED patients (5.8, 4.3-7.5) were significantly higher (p < .001) than in NA-ED patients (4.4, 2.6-5.9) and in control group (4.6, 3.1-7.2). There was a significant difference (p < .001) between uric acid levels in patients with mild A-ED (IIEF-5 16-20) and severe/complete A-ED (IIEF-5 ≤ 10) that were 5.4 (range 4.3-6.5) mg/dL and 6.8 (range 6.4-7.2) mg/dL, respectively. There was no difference between the levels of uric acid in patients with different degree of NA-ED. Our findings reveal that SUA is a marker of ED but only of ED of arteriogenic aetiology.


Assuntos
Impotência Vasculogênica/sangue , Ácido Úrico/sangue , Adulto , Estudos de Casos e Controles , Endotélio Vascular/fisiopatologia , Humanos , Impotência Vasculogênica/diagnóstico , Impotência Vasculogênica/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Ereção Peniana/fisiologia , Inquéritos e Questionários , Ultrassonografia Doppler em Cores
2.
Medicine (Baltimore) ; 94(42): e1754, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26496293

RESUMO

There are no studies comparing some of the most important markers, such as vitamin D, parathormone, osteocalcin, bone alkaline phosphatase, and calcium, in patients with chronic benign and malignant pancreatic diseases. Our objective was to comparatively evaluate serum markers of bone metabolism in patients with chronic pancreatitis and in those with ductal pancreatic adenocarcinoma. Sixty-three consecutive subjects were studied: 30 patients with a firm diagnosis of chronic pancreatitis and 33 having histologically confirmed pancreatic adenocarcinoma. Serum 25-hydroxyvitamin D, bone alkaline phosphatase, osteocalcin, parathormone, and calcium were determined using commercially available kits. Taking into consideration the clinical variables of all 63 patients studied, 25-hydroxyvitamin D was inversely correlated with only the body mass index (P = 0.007), whereas it was not correlated with age (P = 0.583) or fecal elastase-1 concentrations (P = 0.556). Regarding the other substances studied, parathormone was positively correlated with only the age of the patients (P = 0.015). Of the 5 substances studied, only bone alkaline phosphates were significantly different (P < 0.001) between patients with chronic pancreatitis and those with pancreatic ductal adenocarcinoma. Within the 2 groups of patients, the 23 patients with chronic pancreatitis without diabetes mellitus had serum concentrations of 25-hydroxyvitamin D significantly lower (P = 0.045) than those with chronic pancreatitis having diabetes mellitus, whereas smokers with pancreatic ductal adenocarcinoma had serum concentrations of calcium significantly higher (P < 0.001) as compared to nonsmokers. Altered bone metabolism seems to be associated with chronic diseases of the pancreas; however, the mechanism should be better elucidated.


Assuntos
Osso e Ossos/metabolismo , Carcinoma Ductal Pancreático/sangue , Neoplasias Pancreáticas/sangue , Vitamina D/análogos & derivados , Fatores Etários , Idoso , Consumo de Bebidas Alcoólicas , Índice de Massa Corporal , Cálcio/sangue , Carcinoma Ductal Pancreático/complicações , Doença Crônica , Insuficiência Pancreática Exócrina/complicações , Feminino , Humanos , Medições Luminescentes , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/complicações , Hormônio Paratireóideo/sangue , Fumar/epidemiologia , Vitamina D/sangue
3.
Clin Chim Acta ; 438: 252-4, 2015 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-25218698

RESUMO

BACKGROUND: Mid-infrared spectral technology has shown a high degree of promise in detecting glucose in plasma. OptiScan Biomedical has developed a glucose monitor based on mid-infrared spectroscopy that withdraws blood samples and measures plasma glucose. The objective of this study was to evaluate the accuracy and performance of the OptiScanner™ 5000 system on different pools of blood. METHODS: This study was performed to validate the blood glucose measurements obtained with the OptiScanner™ 5000 by comparing them to Central Laboratory glucose measurements (VITROS® 5600 Integrated System) as a comparative method across a broad range of glucose values over a three day period to obtain 80-90 paired measurements. RESULTS: A total of 81 paired measurements, distributed between 33 and 320mg/100mL of glucose, were performed. The aggregate data points were within International Organization for Standardization standards, with 100% of the glucose values within ±15%. CONCLUSIONS: The current study suggests that a mid-IR fixed-wavelength system (OptiScanner) can measure glucose accurately across a wide range of glucose values in plasma of ICU patients.


Assuntos
Glicemia/análise , Espectrofotometria Infravermelho/normas , Humanos , Monitorização Fisiológica/instrumentação , Reprodutibilidade dos Testes , Espectrofotometria Infravermelho/instrumentação , Espectrofotometria Infravermelho/métodos
4.
J Sex Med ; 11(11): 2792-800, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25092061

RESUMO

INTRODUCTION: Endothelial dysfunction has been demonstrated to play an important role in pathogenesis of erectile dysfunction (ED) and vitamin D deficiency is deemed to promote endothelial dysfunctions. AIM: To evaluate the status of serum vitamin D in a group of patients with ED. METHODS: Diagnosis and severity of ED was based on the IIEF-5 and its aetiology was classified as arteriogenic (A-ED), borderline (BL-ED), and non-arteriogenic (NA-ED) with penile-echo-color-Doppler in basal condition and after intracaversous injection of prostaglandin E1. Serum vitamin D and intact PTH concentrations were measured. MAIN OUTCOME MEASURES: Vitamin D levels of men with A-ED were compared with those of male with BL-ED and NA-ED. RESULTS: Fifty patients were classified as A-ED, 28 as ED-BL and 65 as NA-ED, for a total of 143 cases. Mean vitamin D level was 21.3 ng/mL; vitamin D deficiency (<20 ng/mL) was present in 45.9% and only 20.2% had optimal vitamin D levels. Patients with severe/complete-ED had vitamin D level significantly lower (P = 0.02) than those with mild-ED. Vitamin level was negatively correlated with PTH and the correlation was more marked in subjects with vitamin D deficiency. Vitamin D deficiency in A-ED was significantly lower (P = 0.01) than in NA-ED patients. Penile-echo-color-Doppler revealed that A-ED (PSV ≤ 25 cm/second) was more frequent in those with vitamin D deficiency as compared with those with vitamin >20 ng/dL (45% vs. 24%; P < 0.05) and in the same population median PSV values were lower (26 vs. 38; P < 0.001) in vitamin D subjects. CONCLUSION: Our study shows that a significant proportion of ED patients have a vitamin D deficiency and that this condition is more frequent in patients with the arteriogenic etiology. Low levels of vitamin D might increase the ED risk by promoting endothelial dysfunction. Men with ED should be analyzed for vitamin D levels and particularly to A-ED patients with a low level a vitamin D supplementation is suggested.


Assuntos
Disfunção Erétil/etiologia , Deficiência de Vitamina D/complicações , Vitamina D/sangue , Adulto , Disfunção Erétil/sangue , Disfunção Erétil/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Pênis/fisiopatologia , Deficiência de Vitamina D/sangue
6.
Can J Cardiol ; 28(5): 537-41, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22552174

RESUMO

BACKGROUND: Subclinical inflammation and atrial stretch have been recognized as important contributors to atrial fibrillation (AF) onset and perpetuation. The aim of the study was to compare the predictive role of serum inflammatory markers (serum amyloid A [SAA], and C-reactive protein [CRP]) and N-terminal pro brain natriuretic peptide (NT-proBNP) an indice of atrial strain in relation to subacute arrhythmic recurrence rate in patients with persistent AF and normal left ventricular ejection fraction (LVEF). METHODS: We studied 57 patients with a mean LVEF of 58.7 ± 6%. NT-proBNP, SAA and CRP levels were determined few hours before electrical cardioversion and 3 weeks after cardioversion. RESULTS: Subacute AF recurrences were documented in 19 (33 %) patients. Whereas NT-proBNP levels did not predict arrhythmic outcome, higher SAA (> 6.16-6.19 mg/L) and CRP levels (> 2.99-3.10 mg/L) were significantly associated with AF recurrences (odds ratio [OR], 5.39; 95% confidence interval [CI], 1.59-18.26; P = 0.007 and OR, 14.93; 95% CI, 3.90-57.19; P < 0.001). Both SAA (OR, 18.29; 95% CI, 2.07-161.46; P = 0.009) and high sensitivity CRP (OR, 42.03; 95% CI, 4.83-365.45; P = 0.001) through the multivariate logistic regression analysis show an independent role in predicting the AF recurrence with a sensitivity of 100% (38/38) and a specificity of 52.6% (10/19). CONCLUSIONS: The present study demonstrates that in patients with persistent AF and preserved LVEF, SAA and CRP levels are independent predictors of AF subacute recurrence rate, whereas NT-proBNP, not associated with arrhythmic outcome, reflects the hemodynamic alterations secondary to arrhythmia presence. The simultaneous determination of SAA and high sensitivity CRP has a very high sensitivity (100%) in predicting the AF recurrence.


Assuntos
Fibrilação Atrial/sangue , Fibrilação Atrial/terapia , Proteína C-Reativa/metabolismo , Cardioversão Elétrica/efeitos adversos , Proteína Amiloide A Sérica/metabolismo , Idoso , Análise de Variância , Fibrilação Atrial/diagnóstico , Fibrilação Atrial/mortalidade , Estudos de Coortes , Intervalos de Confiança , Cardioversão Elétrica/métodos , Eletrocardiografia , Feminino , Humanos , Mediadores da Inflamação/sangue , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Peptídeo Natriurético Encefálico/sangue , Razão de Chances , Fragmentos de Peptídeos/sangue , Valor Preditivo dos Testes , Prognóstico , Estudos Prospectivos , Recidiva , Medição de Risco , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Taxa de Sobrevida , Função Ventricular Esquerda/fisiologia
7.
J Nephrol ; 25(6): 1047-53, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22362550

RESUMO

INTRODUCTION: We measured serum creatinine concentrations in 17 male athletes of the Italian national rugby team. METHODS: Blood was obtained before the start of training and during the competitive season. Serum creatinine level was measured by Jaffé reaction at 4 time points during the season, with a formal measure of creatinine clearance in midseason. RESULTS: The values of estimated glomerular filtration rate (eGFR) calculated with the Cockcroft-Gault (CG) equation were higher than those calculated with the Modification of Diet in Renal Disease (MDRD) Study formula (p<0.001). This difference was significantly decreased but still present when the MDRD formula was corrected for body surface area (BSA). When compared with measured creatinine clearance (CrCl), the MDRD underestimates the CrCl by 51 ml/min (95% confidence interval [95% CI], 36-67 ml/min, p<0.0001). When corrected for BSA, this difference falls to 27 ml/min (95% CI, 13-44 ml/min, p=0.001). The CG eGFR gave a better estimate of CrCl, differing by 1 ml/min (95% CI, -16 to +17 ml/min, p=NS). CONCLUSIONS: The MDRD formula underestimates the CrCl in rugby players, even when corrected for BSA. Conversely, the CG formula more closely approximates the actual CrCl measurement. The equations to estimate GFR should be used with caution in subjects having atypical anthropometric characteristics.


Assuntos
Creatinina/sangue , Futebol Americano , Taxa de Filtração Glomerular , Rim/fisiologia , Modelos Biológicos , Adulto , Análise de Variância , Biomarcadores/sangue , Superfície Corporal , Nível de Saúde , Humanos , Itália , Rim/metabolismo , Masculino , Fatores de Tempo , Adulto Jovem
8.
Amyloid ; 18(4): 216-21, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22070520

RESUMO

BACKGROUND: Cardiac biomarkers play a major role in the identification of patients at risk of early death in AL amyloidosis, and a staging system based on amino-terminal pro-natriuretic peptide type-B (NT-proBNP) and troponins (cTn) is used for prognostic stratification. Adrenomedullin is produced by several tissues including the heart, and portends a poor prognosis in heart diseases. We investigated the ability of midregional proadrenomedullin (MR-proADM) to predict early death in AL amyloidosis. METHODS: One-hundred and thirty consecutive patients with newly-diagnosed AL amyloidosis were prospectively enrolled. The impact on survival of NT-proBNP, cTnI and MR-proADM was evaluated. RESULTS: The concentration of MR-proADM correlated with systolic and diastolic function, but did not reflect the amount of amyloid deposited in the heart. Moreover, MR-proADM was associated with non-cardiac markers of advanced disease. The staging system based on NT-proBNP and cTnI identified high-risk subjects, but could not discriminate good-risk and intermediate-risk patients. Conversely, a staging system based on MR-proADM and cTnI identified 3 groups with significantly different survivals. CONCLUSIONS: Midregional-proADM is a powerful prognostic marker in AL amyloidosis, which may not only reflect cardiac dysfunction but also widespread systemic disease, and can be combined with cTn for detecting patients at risk of early death.


Assuntos
Adrenomedulina/metabolismo , Amiloide/metabolismo , Amiloidose/diagnóstico , Precursores de Proteínas/metabolismo , Idoso , Amiloidose/sangue , Amiloidose/complicações , Amiloidose/mortalidade , Biomarcadores , Feminino , Insuficiência Cardíaca/sangue , Insuficiência Cardíaca/etiologia , Insuficiência Cardíaca/mortalidade , Ventrículos do Coração/diagnóstico por imagem , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Prognóstico , Modelos de Riscos Proporcionais , Troponina I/sangue , Ultrassonografia
9.
Ther Drug Monit ; 33(3): 303-8, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21544015

RESUMO

BACKGROUND: Adequate plasma trough concentrations (Ctrough) of protease inhibitors are required to maintain antiviral activity throughout the dosing interval. Therapeutic drug monitoring is used in clinical practice to optimize dosage and avoid toxic or subtherapeutic drug exposure. The pharmacokinetic variability of Atazanavir (ATV) can be relatively large, as a result of several factors. One of the affecting factors may be hepatic impairment due to hepatitis C virus (HCV) coinfection. METHODS: We collected trough plasma samples from human immunodeficiency virus (HIV)-1-infected outpatients, with and without HCV coinfection and/or cirrhosis, receiving stable highly active antiretroviral therapy containing ATV. In the total population, we mainly compared the 2 regimens: 300ATV + 100RTV OD [ritonavir (RTV), once daily (OD)] versus 400ATV OD. We used a threshold value of 0.15 µg/mL, based on the proposed therapeutic range (0.15-0.85 µg/mL). Plasma concentrations of ATV were determined by a validated assay using high-performance liquid chromatography with ultraviolet detection. A total of 214 HIV-infected outpatients were included. For each regimen, we compared 3 groups of subjects: HIV+/HCV-, HIV+/HCV+, and HIV+/HCV+ with cirrhosis. RESULTS: In the whole study population, we observed a large variability and found suboptimal Ctrough levels (<0.15 µg/mL) in 23 subjects (2 belonging to the 300/100 OD group and 21 to the 400 OD group). For the standard dosage regimen of 300ATV + 100RTV OD, we did not find a statistical difference between HIV-infected patients without HCV coinfection versus HIV-infected patients with HCV coinfection: median 0.85 (interquartile range 0.53-1.34) and 0.95 (0.70-1.36) µg/mL, respectively. In HIV+/HCV+-infected patients with cirrhosis, we found a median Ctrough of 0.70 (0.43-1.0) µg/mL, with no statistical difference when compared with HIV+/HCV- infected patients. For the 400ATV OD (n=90) dosage regimen, the total median ATV Ctrough was 0.40 (0.23-1.0) µg/mL. In this group, we found a statistically significant difference between HIV+/HCV- and HIV+/HCV+-infected patients: median Ctrough was 0.23 (0.11-0.42) and 0.52 (0.20-1.0) µg/mL, respectively. In HIV+/HCV+ subjects with cirrhosis, the Ctrough median value was 0.42 (0.13-0.75) µg/mL, and there was a significant difference when compared with HIV patients without coinfection. CONCLUSIONS: Therapeutic drug monitoring of ATV in patients receiving unboosted regimen may be useful to identify those HIV-infected subjects, with or without HCV coinfection, who may benefit from adding low RTV doses, or the subset of patients in whom removal of RTV could be attempted without the risk of suboptimal plasma ATV exposure.


Assuntos
Infecções por HIV/sangue , Infecções por HIV/tratamento farmacológico , Inibidores da Protease de HIV/administração & dosagem , Hepatite C/sangue , Oligopeptídeos/administração & dosagem , Oligopeptídeos/sangue , Piridinas/administração & dosagem , Piridinas/sangue , Adulto , Terapia Antirretroviral de Alta Atividade/métodos , Sulfato de Atazanavir , Monitoramento de Medicamentos/métodos , Feminino , Fibrose/sangue , Fibrose/virologia , HIV/isolamento & purificação , Infecções por HIV/virologia , Inibidores da Protease de HIV/sangue , Inibidores da Protease de HIV/farmacocinética , Hepacivirus/isolamento & purificação , Hepatite C/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Oligopeptídeos/farmacocinética , Piridinas/farmacocinética , Estudos Retrospectivos , Ritonavir/uso terapêutico
10.
Blood ; 116(18): 3426-30, 2010 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-20644111

RESUMO

In light-chain (AL) amyloidosis, prognosis is dictated by cardiac dysfunction. N-terminal natriuretic peptide type B (NT-proBNP) and cardiac troponins (cTn) are used to assess the severity of cardiac damage. We evaluated the prognostic relevance of a high-sensitivity (hs) cTnT assay, NT-proBNP, and cardiac troponin I in 171 consecutive patients with AL amyloidosis at presentation and 6 months after treatment. Response and progression of NT-proBNP were defined as more than 30% and more than 300 ng/L changes. All 3 markers predicted survival, but the best multivariable model included hs-cTnT. The hs-cTnT prognostic cutoff was 77 ng/L (median survival 10.6 months for patients with hs-cTnT above the cutoff). After treatment, response and progression of NT-proBNP and a more than 75% increase of hs-cTnT were independent prognostic determinant. In AL amyloidosis, hs-cTnT is the best baseline prognostic marker. Therapy should be aimed at preventing progression of cardiac biomarkers, whereas NT-proBNP response confers an additional survival benefit.


Assuntos
Amiloidose/diagnóstico , Peptídeo Natriurético Encefálico , Fragmentos de Peptídeos , Troponina T , Adulto , Idoso , Idoso de 80 Anos ou mais , Amiloidose/tratamento farmacológico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miocárdio/patologia , Prognóstico , Análise de Sobrevida , Troponina I
11.
Clin Chem ; 55(3): 499-504, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19131635

RESUMO

BACKGROUND: The diagnosis of systemic immunoglobulin light-chain (AL) amyloidosis requires demonstration of amyloid deposits in a tissue biopsy and amyloidogenic monoclonal light chains. The optimal strategy to identify the amyloidogenic clone has not been established. We prospectively assessed the diagnostic sensitivity of the serum free light chain (FLC) kappa/lambda ratio, a commercial serum and urine agarose gel electrophoresis immunofixation (IFE), and the high-resolution agarose gel electrophoresis immunofixation (HR-IFE) developed at our referral center in patients with AL amyloidosis, in whom the amyloidogenic light chain was unequivocally identified in the amyloid deposits. METHODS: The amyloidogenic light chain was identified in 121 consecutive patients with AL amyloidosis by immunoelectron microscopy analysis of abdominal fat aspirates and/or organ biopsies. We characterized the monoclonal light chain by using IFE and HR-IFE in serum and urine and the FLC kappa/lambda ratio in serum. We then compared the diagnostic sensitivities of the 3 assays. RESULTS: The HR-IFE of serum and urine identified the amyloidogenic light chain in all 115 patients with a monoclonal gammopathy. Six patients with a biclonal gammopathy were omitted from the statistical analysis. The diagnostic sensitivity of commercial serum and urine IFE was greater than that of the FLC kappa/lambda ratio (96% vs 76%). The combination of serum IFE and the FLC assay detected the amyloidogenic light chain in 96% of patients. The combination of IFE of both serum and urine with the FLC kappa/lambda ratio had a 100% sensitivity. CONCLUSIONS: The identification of amyloidogenic light chains cannot rely on a single test and requires the combination of a commercially available FLC assay with immunofixation of both serum and urine.


Assuntos
Amiloidose/sangue , Amiloidose/urina , Imunoensaio/métodos , Cadeias Leves de Imunoglobulina/sangue , Cadeias Leves de Imunoglobulina/urina , Amiloidose/imunologia , Biomarcadores/sangue , Biomarcadores/urina , Feminino , Humanos , Cadeias Leves de Imunoglobulina/imunologia , Masculino
12.
J Cardiovasc Med (Hagerstown) ; 9(6): 581-8, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18475126

RESUMO

OBJECTIVES: Maintenance of sinus rhythm after cardioversion of atrial fibrillation is a major clinical challenge also in patients with preserved left ventricular function. Subclinical inflammation and atrial strain have been recognized as important contributors to atrial fibrillation onset and perpetuation. Aim of the study was to compare the predictive role of C-reactive protein (CRP) and indices of atrial dysfunction in relation to subacute arrhythmic recurrence rate in patients with persistent atrial fibrillation and normal left ventricular ejection fraction (LVEF). METHODS: We studied 53 patients with a mean LVEF of 58.7 +/- 6%. Left atrial diameter and area, left atrial auricle emptying velocity, N-terminal pro-b-type natriuretic peptide (NT-proBNP) and CRP levels were determined few hours before electrical cardioversion. NT-proBNP and CRP levels were also measured 1 h and 3 weeks after cardioversion. RESULTS: Subacute atrial fibrillation recurrences were documented in 18 (33.9%) patients. Whereas none of the parameters reflecting atrial dysfunction predicted arrhythmic outcome, higher CRP levels (>3.0 mg/l) were significantly associated with atrial fibrillation recurrences [odds ratio (OR): 1.6; 95% confidence interval (CI): 1.4-2.5; P = 0.031]. No changes in CRP levels were evident after cardioversion independently of underlying rhythm. On the contrary, NT-proBNP levels, which were correlated with left atrial auricle emptying velocity, significantly decreased only in patients who maintained sinus rhythm (from 638 +/- 329 to 295 +/- 261 pg/ml; P < 0.001). CONCLUSION: The present study demonstrates that in patients with persistent atrial fibrillation and preserved LVEF, CRP level is an independent predictor of atrial fibrillation subacute recurrence rate, whereas none of the indices of atrial dysfunction is associated with arrhythmic outcome. NT-proBNP levels reflect, instead, the hemodynamic alterations secondary to arrhythmia presence.


Assuntos
Fibrilação Atrial/diagnóstico , Fibrilação Atrial/terapia , Função Atrial/fisiologia , Biomarcadores/sangue , Proteína C-Reativa/análise , Cardioversão Elétrica , Volume Sistólico/fisiologia , Idoso , Fibrilação Atrial/fisiopatologia , Feminino , Humanos , Masculino , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Recidiva
13.
J Mol Diagn ; 9(4): 538-45, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17652639

RESUMO

To quantify the amount of the 3460G-->A/ND1 point mutation responsible for Leber's hereditary optic neuropathy, we developed a quantitative real-time polymerase chain reaction method based on the SYBR Green assay and a new approach using the TaqMan assay. Both methods were based on the amplification refractory mutation system, comparing the heteroplasmic load quantified by restriction fragment length polymorphism in 15 Leber's hereditary optic neuropathy family members, with the results obtained using quantitative real-time polymerase chain reaction methods. The comparative evaluation of mitochondrial DNA (mtDNA) heteroplasmy from blood samples showed significant correlation between restriction fragment length polymorphism analysis, real-time SYBR Green assay, and TaqMan assay. We validated the last method by measuring experimental samples composed by a known proportion of cloned plasmids containing either the wild-type or mutant sequence, giving a correlation coefficient of 0.999 (P < 0.0001). The real-time amplification refractory mutation system polymerase chain reaction by TaqMan assay provides a rapid, reliable, sensitive, reproducible, and one-step quantitative method to detect heteroplasmic mutant mtDNA. This method allows the quantitation of a broad range of mutational load (up to 100%, down to 0.01%) on the basis of in vitro calibration, thus rendering the TaqMan assay suitable for the diagnostic analysis of heteroplasmic load in mtDNA-related disorders.


Assuntos
DNA Mitocondrial/genética , Atrofia Óptica Hereditária de Leber/genética , Mutação Puntual/genética , Reação em Cadeia da Polimerase/métodos , Calibragem , Sondas de DNA/metabolismo , Feminino , Corantes Fluorescentes/metabolismo , Humanos , Masculino , Linhagem
15.
Clin Chem Lab Med ; 43(8): 883-5, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16201902

RESUMO

We evaluated the analytical performance of a new, commercial, fully automated immunoturbidimetric assay for the determination of ferritin [FER-Latex(X2)CN SEIKEN, Denka Seiken, Japan] in serum on the Olympus AU2700 analyzer. The new assay is a latex-enhanced turbidimetric immunoassay with an analysis time of 10 min. The linearity of the assay was confirmed up to 2505 pmol/L (R2=0.999). The detection limit and the functional sensitivity were both 4.5 pmol/L. The intra- and inter-assay imprecision (CV) at 67, 506, 2186 pmol/L was < 1.8% and < 2.5%, respectively. Verification of the traceability to a WHO standard (80/578) showed a recovery of 102.6% (target value 449 pmol/L). No hook effect was observed in samples containing up to 33,705 pmol/L. The assay showed good correlation with the Beckman Immage nephelometric system (r=0.999). Hemoglobin (< or = 9.8 g/L), total bilirubin (< or = 113 micromol/L), conjugated bilirubin (< or = 109 micromol/L) and rheumatoid factor (< or = 5.2x10(5) IU/L) did not interfere with the assay. The reference interval (2.5-97.5 percentile) was 72-521 pmol/L for men and 27-267 pmol/L for women. The reference interval in patients with anemia, malignant tumors and hemochromatosis was 5.6-52, 130-2436 and 1465-2903 pmol/L, respectively. On the basis of the receiver operating characteristic curve, the 90% sensitivity cut-off value to distinguish between patients with and without iron deficiency was 40 pmol/L. The new latex turbidimetric procedure for ferritin assay is an attractive alternative that avoids the need for dedicated instrumentation.


Assuntos
Análise Química do Sangue/métodos , Análise Química do Sangue/estatística & dados numéricos , Ferritinas/sangue , Imunoensaio/métodos , Imunoensaio/estatística & dados numéricos , Adulto , Anemia Ferropriva/sangue , Análise Química do Sangue/normas , Feminino , Ferritinas/normas , Hemocromatose/sangue , Humanos , Imunoensaio/normas , Masculino , Pessoa de Meia-Idade , Neoplasias/sangue , Curva ROC , Padrões de Referência , Valores de Referência , Sensibilidade e Especificidade
16.
Biochem J ; 384(Pt 2): 287-93, 2004 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-15324306

RESUMO

We analysed key biochemical features that reflect the balance between glycolysis and glucose oxidation in cybrids (cytoplasmic hybrids) harbouring a representative sample of mitochondrial DNA point mutations and deletions. The cybrids analysed had the same 143B cell nuclear background and were isogenic for the mitochondrial background. The 143B cell line and its rho(0) counterpart were used as controls. All cells analysed were in a dynamic state, and cell number, time of plating, culture medium, extracellular volume and time of harvest and assay were strictly controlled. Intra- and extra-cellular lactate and pyruvate levels were measured in homoplasmic wild-type and mutant cells, and correlated with rates of ATP synthesis and O2 consumption. In all mutant cell lines, except those with the T8993C mutation in the ATPase 6 gene, glycolysis was increased even under conditions of low glucose, as demonstrated by increased levels of extracellular lactate and pyruvate. Extracellular lactate levels were strictly and inversely correlated with rates of ATP synthesis and O2 consumption. These results show increased glycolysis and defective oxidative phosphorylation, irrespective of the type or site of the point mutation or deletion in the mitochondrial genome. The different biochemical consequences of the T8993C mutation suggest a uniquely different pathogenic mechanism for this mutation. However, the distinct clinical features associated with some of these mutations still remain to be elucidated.


Assuntos
Respiração Celular/fisiologia , Citoplasma/química , DNA Mitocondrial/genética , Células Híbridas/química , Mutação/genética , Trifosfato de Adenosina/biossíntese , Plaquetas/química , Plaquetas/metabolismo , Linhagem Celular , Ácido Cítrico/metabolismo , Citoplasma/metabolismo , Matriz Extracelular/química , Fibroblastos/química , Fibroblastos/metabolismo , Humanos , Células Híbridas/metabolismo , Espaço Intracelular/química , Ácido Láctico/metabolismo , Mioblastos/química , Mioblastos/metabolismo , Consumo de Oxigênio/fisiologia , Ácido Pirúvico/metabolismo
17.
J Pediatr Gastroenterol Nutr ; 36(3): 392-6, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12604981

RESUMO

UNLABELLED: Fecal elastase is considered to be a highly sensitive and specific non-invasive exocrine pancreatic function test. However, enteropathy may theoretically cause decreased exocrine pancreatic enzyme secretion through alteration of enteric hormone release. OBJECTIVE: The aim of this study was to evaluate the possible influence of transient small bowel damage on pancreatic elastase secretion. METHODS: We studied 166 children (aged 4 months to 14 years, mean 2 years); 114 of these children had acute enteritis and 52 children were control subjects (with gastro-intestinal symptoms or extra-intestinal diseases). Feces were collected from each patient 3 days after the onset of diarrhea and then tested for fecal elastase, bacterial pathogens, Rotavirus, and Adenovirus. Liquid fecal samples were not considered eligible for elastase measurement. Pancreatic elastase was measured using an ELISA method (Sche.Bo.Tech, Germany). We classified the results, expressed in microg/g stool, as: severe pancreatic insufficiency (<100 microg/g), moderate pancreatic insufficiency (100 to 200 microg/g), and normal (>200 microg/g). RESULTS: In the acute enteritis group we found severe levels in 14 (12%) children, moderate levels in 18 children (16%), and normal levels in 82 children (72%). In contrast, 52 of 52 (100%) control subjects demonstrated normal results. Statistical analysis (Wilcoxon rank test) demonstrated a significant difference between the enteritis and control groups (P < 0.01). Serial measurement of fecal elastase performed in 10 patients with enteritis showed a progressive increase of levels in 6 patients and an early decline with subsequent increases in the other 4 patients. CONCLUSIONS: Transient exocrine pancreatic insufficiency may be present in transient small bowel disease, caused by both bacterial and viral infections, possibly related to reduced enteric CCK secretion.


Assuntos
Enterite/fisiopatologia , Insuficiência Pancreática Exócrina/diagnóstico , Fezes/enzimologia , Intestino Delgado/patologia , Pâncreas/fisiologia , Elastase Pancreática/análise , Testes de Função Pancreática , Doença Aguda , Adolescente , Biomarcadores/análise , Criança , Pré-Escolar , Enterite/microbiologia , Enterite/virologia , Ensaio de Imunoadsorção Enzimática/métodos , Insuficiência Pancreática Exócrina/enzimologia , Feminino , Humanos , Lactente , Masculino , Pâncreas/enzimologia , Testes de Função Pancreática/métodos , Índice de Gravidade de Doença
18.
JOP ; 3(5): 139-43, 2002 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12221328

RESUMO

BACKGROUND: There is a lack of data concerning pancreatic involvement during shock. AIM: To evaluate possible pancreatic alterations in the early phase of shock. SETTING: Twelve consecutive patients with shock were studied within 2 hours from the onset of illness. Seven patients died during the hospital stay: 3 within 4 hours from admission, 3 within 4-8 hours and 1 within 12 hours. MAIN OUTCOME MEASURES: Amylase, lipase, C-reactive protein, amyloid A, interleukin 6, procalcitonin and vascular cell adhesion molecule-1 serum concentrations were determined on admission and 4, 8, and 12 hours afterward. All patients underwent imaging studies of the pancreas. RESULTS: None of the patients developed clinical signs or morphological alterations compatible with acute pancreatitis. Serum amylase levels were above the upper reference limit in 7 patients (58.3%) and serum lipase levels in 2 patients (16.7%; P=0.062). There were no significant differences found between survivors and non-survivors in the serum concentrations of all the proteins studied. CONCLUSIONS: In patients with shock, amylase seems to be more frequently elevated than lipase. None of the patients showed pancreatic alterations at imaging techniques.


Assuntos
Pâncreas/fisiopatologia , Choque/fisiopatologia , Idoso , Idoso de 80 Anos ou mais , Amilases/sangue , Feminino , Humanos , Lipase/sangue , Masculino , Pâncreas/enzimologia , Choque/sangue , Choque/enzimologia , Choque/mortalidade
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