Emerging biomarkers for improving pregnancy planning in multiple sclerosis.

Background: Patient disability, relapse rate, and age are used for family planning in multiple sclerosis (MS). However, the need for more accurate biomarkers is widely recognized. We aimed to explore the influence of age on neurofilament light chain (sNfL), which reflects acute inflammation; glial fibrillary acidic protein (GFAP), associated with disability progression independent of relapses; and anti-Müllerian hormone (AMH), reflecting ovarian reserve, to provide a tailored family planning strategy. Methods: This case-control study included 95 MS patients and 61 healthy control women (HCW). sNfL and GFAP levels were measured using a sensitive single-molecule array assay. AMH levels were measured by the automated Elecsys® Anti-Müllerian Hormone Assay. Results: We observed no significant differences in AMH values between MS patients and the control group within any of the age-matched categories. Age exhibited a negative correlation with AMH values in both groups, as expected. Nevertheless, our findings suggest a slight tendency toward reduced ovarian reserve in MS patients (rho MS patients = -0.67, p < 0.0001; rho HCW = -0.43, p = 0.0006). Interestingly, among the 76 MS participants under 40 years old, we identified ten individuals (13.1%) with AMH levels below 0.7 ng/ml, indicative of a low ovarian reserve, and an additional six individuals (7.8%) with AMH levels between 0.7 ng/ml and 0.9 ng/ml, suggesting a potential risk of premature ovarian failure. Conversely, sNfL and GFAP levels in the MS group exhibited high variability but showed no significant association with age intervals. Conclusion: We found no significant differences in AMH, sNfL or GFAP values between MS patients and the control group within any of the age-matched categories. The assessment of AMH, sNFL and GFAP levels at MS onset facilitates personalized therapeutic and family planning strategies for childbearing-age women.

IL-6 and Other Biomarkers associated with Poor Prognosis in a Cohort of Hospitalized Patients with COVID-19 in Madrid.

OBJECTIVES: There are several published works on the prognostic value of biomarkers in relation to the severity or fatal outcome of coronavirus disease 2019 (COVID-19). In Spain, the second European country in incidence of the disease at the time of data collection, there are few studies that include both laboratory parameters and clinical parameters. Our aim is to study the relationship of a wide series of biomarkers with admission to intensive care and death in a hospital in the Autonomous Community of Madrid (Spain), with special attention to IL-6 due to its role in the systemic inflammatory response associated with a worse prognosis of the disease. METHODS: Data were collected from 546 hospitalized patients with COVID-19. All of them had IL-6 results, in addition to other biochemical and haematological parameters. The difference of the medians for the selected parameters between the groups (ICU vs non-ICU, dead vs survivors) was studied using a Mann-Whitney analysis. The independent variables that predicted death were studied using a Cox proportional hazard regression model. RESULTS: Higher age and blood concentrations of ALT, creatinine, CK, cTnI, LDH, NT-proBNP, CRP, IL-6, leucocyte count and D-dimer together with lower blood concentrations of albumin and lymphocyte count were associated with mortality in univariate analysis. Age, LDH, IL-6 and lymphocyte count remained associated with death in multivariate analysis. CONCLUSIONS: Age, LDH, IL-6 and lymphocyte count, as independent predictors of death, could be used to establish more aggressive therapies in COVID-19 patients.

Comparison of three methods for determining anti-thyrotropin receptor antibodies (TRAb) for diagnosis of Graves' disease: a clinical validation.

Objectives: Graves' disease is secondary to the presence of anti-thyrotropin receptor antibodies (TRAb), which stimulate thyroid hormones. TRab determination is crucial for etiological diagnosis. The objectives of this study were (i) to compare two methods for determining TRab by chemoluminiscence vs. standard TRACE-immunofluorescence; (ii) to determine the diagnostic validity of the three methods. Methods: A retrospective study in 194 patients with a TRAb determination request. TRAb were determined by immunofluorescence (Kryptor, ThermoFisher) and chemiluminescence (Immulite, Siemens and Maglumi, Snibe). Clinical validation: medical records were reviewed and categorized according to thyroid function. Statistical analysis: Differences in quantitative variables were assessed by intraclass correlation coefficient, Bland-Altman plot, and mean differences (mD). Qualitative variables were dichotomized by cut-off points; Kappa coefficient was calculated. Correlations were evaluated by Pearson's coefficient and Passing-Bablok regression analysis. The diagnostic validity of the three methods was investigated. Results: Kryptor-Immulite: mD: 1.2 (95%CI: -16 to >18). Passing-Bablok: Constant error (95%CI: -0.8349 to -0.5987). Proportional error (95%CI: 0.7862-1.0387). ICC: 0.86 (95%CI: 0.82-0.89). Kappa coefficient: 0.68 (95%CI 0.59-0.78). Kryptor-Maglumi: mD: -0.3 (95%CI: -12 to >12). Passing-Bablok: Constant error (95%CI: -0.7701 to >0.1621. Proportional error (95%CI: 0.8571 to 1.3179. ICC: 0.93 (95%CI: 0.89-0.97). Kappa coefficient: 0.53 (95%CI: 0.32-0.74). Diagnosis of Graves' disease was confirmed in 113 patients (Kryptorf showed better specificity and positive predictive value, whereas Immulite demonstrated better sensitivity and negative predictive value). Conclusions: The three methods have a good diagnostic performance for Graves' disease, with superimposable results on Bland-Altman plot. Interchangeability was not confirmed on the regression and agreement analysis, with the presence of biases.

Un análisis general de la prolisil-oxidasa circulante en pacientes no isquémicos con una insuficiencia cardiaca común con fracción de eyección conservada / A global assessment of circulating prolysyl oxidase in nonischemic patients with garden-variety heart failure with preserved ejection fraction

Introducción y objetivos: La enzima lisil-oxidasa se expresa al alza en el miocardio de pacientes con cardiopatía hipertensiva. Se propone investigar si los pacientes con insuficiencia cardiaca y fracción de eyección conservada de origen hipertensivo-metabólico (ICFEc-HM) presentaban también concentraciones elevadas de prolisil-oxidasa circulante (pLOXc) y las posibles consecuencias de ello. Métodos: Se cuantifican las concentraciones de pLOXc de 85 pacientes no isquémicos con ICFEp-HM en estadio C y se comparan con las de 51 controles sanos. Se evaluaron además las correlaciones entre las concentraciones de pLOXc y ciertos parámetros de rigidez miocárdica, productos del ciclo del colágeno y citocinas fibrogénicas, así como el valor predictivo de la concentración plasmática de la proenzima a 1 año de seguimiento. Resultados: Se detectaron valores aumentados de pLOXc y se encontró que se correlacionaban con los cocientes E/E' y las constantes de rigidez que se calcularon. El subgrupo de pacientes con disfunción diastólica de tipo 1 mostró una correlación negativa solo entre la pLOXc y el péptido natriurético cerebral, mientras que en los pacientes con un patrón diastólico restrictivo se demostró una fuerte correlación entre la pLOXc y la galectina-3. El análisis de Kaplan-Meier reveló que las concentraciones de pLOXc > 52,20 ng/ml incrementaron ligeramente el riesgo de desenlace fatal (test de log rank= 4,45; p = 0,034). Al aplicar la regresión de COX, se obtuvo que la pLOXc es un significativo predictor independiente de muerte u hospitalización por descompensación de la ICFEp-HM (HR = 1,360; IC95%, 1,126-1,638; p = 0,046). Conclusiones: Los pacientes con ICFEp-HM sintomática tienen altas concentraciones séricas de pLOXc, lo cual se asocia con índices de llenado diastólico restrictivo. Tales concentraciones representan un factor de riesgo moderado de mal pronóstico. A lo largo de la historia natural de la ICFEp-HM, se ha constatado que las concentraciones de pLOXc al principio se correlacionan negativamente con las de péptido natriurético cerebral, y después tienen correlación positiva con las de galectina-3, a medida que se desarrolla una disfunción diastólica avanzada

Introduction and objectives: Lysyl oxidase is overexpressed in the myocardium of patients with hypertensive cardiomyopathy. We aimed to explore whether patients with hypertensive-metabolic heart failure with preserved ejection fraction (HM-HFpEF) also have increased concentrations of circulating prolysyl oxidase (cpLOX) and its possible consequences. Methods: We quantified cpLOX concentrations in 85 nonischemic patients with stage C, HM-HFpEF, and compared them with those of 51 healthy controls. We also assessed the correlations of cpLOX with myocardial stiffness parameters, collagen turnover products and fibrogenic cytokines, as well as the predictive value of plasma proenzyme levels at 1-year of follow-up. Results: We detected raised cpLOX values and found that they correlated with calculated E/E' ratios and stiffness constants. The subgroup of patients with type I diastolic dysfunction showed a single negative correlation between cpLOX and B-type natriuretic peptide whereas patients with a restrictive diastolic pattern showed a strong correlation between cpLOX and galectin-3. Kaplan-Meier analysis revealed that cpLOX > 52.20 ng/mL slightly increased the risk of a fatal outcome (log-rank = 4.45; P = .034). When Cox regression was used, cpLOX was found to be a significant independent predictor of cardiovascular death or hospitalization due to the decompensation of HM-HFpEF (HR, 1.360; 95%CI, 1.126-1.638; P = .046). Conclusions: Patients with symptomatic HM-HFpEF show high cpLOX serum levels associated with restrictive diastolic filling indices. These levels represent a moderate risk factor for poor clinical outcome. Throughout the natural history of HM-HFpEF, we observed that cpLOX concentrations were initially negatively correlated with B-type natriuretic peptide but positively correlated with galectin-3 as advanced diastolic dysfunction developed

A Global Assessment of Circulating Prolysyl Oxidase in Nonischemic Patients With Garden-variety Heart Failure With Preserved Ejection Fraction.

INTRODUCTION AND OBJECTIVES: Lysyl oxidase is overexpressed in the myocardium of patients with hypertensive cardiomyopathy. We aimed to explore whether patients with hypertensive-metabolic heart failure with preserved ejection fraction (HM-HFpEF) also have increased concentrations of circulating prolysyl oxidase (cpLOX) and its possible consequences. METHODS: We quantified cpLOX concentrations in 85 nonischemic patients with stage C, HM-HFpEF, and compared them with those of 51 healthy controls. We also assessed the correlations of cpLOX with myocardial stiffness parameters, collagen turnover products and fibrogenic cytokines, as well as the predictive value of plasma proenzyme levels at 1-year of follow-up. RESULTS: We detected raised cpLOX values and found that they correlated with calculated E/E' ratios and stiffness constants. The subgroup of patients with type I diastolic dysfunction showed a single negative correlation between cpLOX and B-type natriuretic peptide whereas patients with a restrictive diastolic pattern showed a strong correlation between cpLOX and galectin-3. Kaplan-Meier analysis revealed that cpLOX > 52.20 ng/mL slightly increased the risk of a fatal outcome (log-rank = 4.45; P = .034). When Cox regression was used, cpLOX was found to be a significant independent predictor of cardiovascular death or hospitalization due to the decompensation of HM-HFpEF (HR, 1.360; 95%CI, 1.126-1.638; P = .046). CONCLUSIONS: Patients with symptomatic HM-HFpEF show high cpLOX serum levels associated with restrictive diastolic filling indices. These levels represent a moderate risk factor for poor clinical outcome. Throughout the natural history of HM-HFpEF, we observed that cpLOX concentrations were initially negatively correlated with B-type natriuretic peptide but positively correlated with galectin-3 as advanced diastolic dysfunction developed.

Regulation of endothelin-converting enzyme-1 (ECE-1) by the calcimimetic R-568.

Although calcimimetics were developed to block parathyroid hormone synthesis, some reports suggest that they may also reduce blood pressure by unknown mechanisms. Calcimimetic-induced changes in the synthesis of endothelial vasoactive factors could be involved. Wistar rats were treated with the calcimimetic R-568, and systolic blood pressure (SBP) was registered with a tail-cuff sphygmomanometer, the content of endothelial nitric oxide synthase (eNOS) and endothelin-converting enzyme (ECE-1) in tissue was evaluated by immunohistochemistry and Western blot, circulating levels of endothelin-1 (ET-1) were measured by ELISA. R-568 reduced SBP and circulating levels of ET-1, without changes in eNOS expression. In contrast, R-568 increased the lung and vascular content of ECE-1. In order to analyze the mechanisms involved, we studied the effect of R-568 on human endothelial cells. R-568 did not modify neither eNOS protein content nor pre-pro-ET-1 mRNA expression, but increased ECE-1 protein content, and decreased ET-1 synthesis and ECE-1 activity. The inhibition of ECE-1 activity was very strong, similar to the classic ECE inhibitor phosphoramidon, the addition of exogenous zinc restored enzymatic activity. Moreover, the amount of zinc in immunoprecipitated ECE from R-568 treated cells was 3-fold less than in control cells. In conclusion, R-568 inhibits ECE by expelling zinc from the enzyme, with the subsequent decrease in enzymatic activity and reducing circulating levels of ET-1, which may be responsible for the lower SBP observed in R-568-treated rats. This descent would be partially compensated by the increased synthesis of the ECE-1 itself, and by other homeostatic mechanisms that regulate SBP.

Evaluation of functional integrity of the retinohypothalamic tract in advanced glaucoma using multifocal electroretinography and light-induced melatonin suppression.

The aim of the study was to investigate the survival of melanopsin-expressing retinal ganglion cells (mRGCs) and the functional integrity of the retinohypothalamic tract in patients with bilateral advanced glaucomatous optic neuropathy by measuring the neuroendocrine light response of the pineal gland. Nine patients with bilateral advanced primary open-angle glaucoma (glaucoma group) and nine normal control subjects (control group) were included in this pilot observational, prospective, case-control study. The best-corrected visual acuity logMAR, standard automated perimetry mean deviation, and the retinal nerve fiber layer thickness determined by optical coherence tomography and multifocal electroretinography were used to evaluate the changes. Melatonin was analyzed in the saliva by radioimmunoassay before and after exposure to bright light (600 lux) for 60 min at night. The advanced glaucoma group did not have any significant nocturnal melatonin suppression after exposure to bright light (14.28 ± 3.07 pg/ml pre-light melatonin concentration vs. 15.22 ± 3.56 pg/ml after light exposure; p = 0.798) unlike the marked melatonin suppression in the control group (22.43 ± 4.37 pg/ml pre-light melatonin concentration vs. 11.25 ± 1.89 pg/ml after light exposure; p < 0.002). Response density estimates by the scalar product amplitude measure for the interval 0-80 ms of the first-order kernel responses were similar in both groups, indicating that outer retinal function was significantly unchanged in the glaucoma group (5.95 ± 0.54 nV/dg^2) compared with the control group (6.20 ± 0.22 nV/dg^2) (p = 0.689). Our findings are consistent with the interpretation that the rhythmic secretion of melatonin was affected in advanced glaucoma, suggesting that attention should be paid to non-image-forming visual functions, such as control of circadian rhythm and the clinical impact in patients with glaucoma.

Procedimiento de validación de magnitudes bioquímicas en un gasómetro. Aplicación al alcance flexible en la Norma ISO 15189 / Validation procedure of biochemical parameters in a gasometer. Application to the flexible scope in the standard ISO 15189

Objetivos. Diseñar un procedimiento de validación de 4 magnitudes bioquímicas en un gasómetro frente a un analizador acreditado que sea sencillo, consistente y aceptable según la Norma ISO 15189:2007 y que pueda servir como método de acreditación flexible. Métodos. Medición secuencial en los analizadores Dimension RxL y GEM 4000 de glucosa, sodio (Na+), potasio (K+) y lactato en 91 muestras de plasma (heparina de litio) de pacientes entre marzo y junio de 2009 (EP-9). Se chequearon outliers y error sistemático. Los pares de resultados se estudiaron mediante análisis de regresión lineal y gráficos de Bland-Altman. Métodos. El criterio de aceptación fue obtener un error sistemático inferior a las especificaciones de calidad definidas en el laboratorio acreditado. Resultados y discusión. Los resultados de glucosa y K+ fueron aceptables según el procedimiento 6.1 de la guía EP-9, mientras que Na+ fue aceptable según el procedimiento 6.2. En cuanto a lactato, se detectó un error sistemático superior a la especificación del laboratorio. Conclusiones. Se han validado los métodos de determinación de glucosa, Na+ y K+ en un gasómetro GEM 4000 mediante la aplicación de la norma EP-9 con respecto a los del Dimension RxL (métodos de referencia). El lactato no se pudo validar porque el error sistemático superó la especificación del laboratorio. Conclusiones. Este procedimiento se propone como herramienta de validación de métodos para laboratorios acreditados con alcance flexible según la Norma UNE EN ISO 15189 (AU)

Objectives. Designing a 4 biochemical parameters validation procedure in a gasometer versus an accredited analyzer. This procedure is simple, robust and acceptable according to the standard ISO 15189: 2007 and that can serve as flexible accreditation method. Methods. Sequential measurement in Dimension RxL and GEM 4000 analysers of glucose, Na+, K+ and lactate in 91 patients plasma samples (Lithium heparin) between March and June 2009 (EP-9). Checked outliers and bias results pairs were studied using linear regression analysis and Bland-Altman graphics. Methods. The acceptance criterion was to get a bias lower than quality specifications defined in the accredited laboratory. Results and discussion. The results of glucose and K+ were acceptable, following the procedure 6.1 of EP9 guideline, while for Na+ it was acceptable by the procedure 6.2. For lactate, a superior to the specification of the lab bias was detected. Conclusions. Measuring methods of glucose, sodium and potassium in a GEM 4000 gasometer applying standard EP9 versus Dimension RxL as reference methods have been validated. Lactate could not be validated because the bias exceeded the specification of the laboratory. Conclusions. This procedure is intended as validation tool of methods for laboratories accredited with flexible scope according to UNE EN ISO 15189 standard (AU)

[Serum PTH levels as a predictive factor of hypocalcaemia after total thyroidectomy]. / Determinación de paratirina en suero como factor predictivo de hipocalcemia tras tiroidectomía total.

INTRODUCTION: Postoperative parathyroid hormone (PTH) levels as a predictor of hypocalcaemia in patients subjected to total thyroidectomy is analyzed. MATERIAL AND METHOD: Prospective study involving 67 patients who underwent total thyroidectomy due to a benign disease. Serum PTH and ionised calcium were measured 20 h after surgery. Sensitivity, specificity and predictive values of PTH and ionised calcium levels were calculated to predict clinical and analytical hypocalcaemia. RESULTS: A total of 42 (62.7%) patients developed hypocalcaemia (ionised calcium<0.95 mmol/l), but only 20 (29.9%) presented with symptoms. PTH concentration the day after surgery was significantly lower in the group that developed symptomatic hypocalcaemia (5.57+/-6.4 pg/ml) than in the asymptomatic (21.5+/-15.3 pg/ml) or normocalcaemic (26.8+/-24.9 pg/ml) groups (p=0.001). Taking the value of 13 pg/ml as a cut-off point of PTH levels, sensitivity, specificity, positive predictive value and negative predictive value were 54%, 72%, 76% and 48%, respectively. On the other hand, sensitivity for predicting symptomatic hypocalcaemia was 95% and specificity was 76%. The test showed a high incidence of false positives (11/30, 36%). Negative predictive value was 97% and positive predictive value was 65%. In multivariate analysis, PTH and ionised calcium were the only perioperative factors that showed an independent predictive value as risk indicators of symptomatic hypocalcaemia. CONCLUSIONS: Normal PTH levels 20 h after surgery practically rule out the subsequent appearance of hypocalcaemia symptoms. On the other hand, low PTH levels are not necessarily associated to symptomatic hypocalcaemia due to the high number of false positives.

Determinación de paratirina en suero como factor predictivo de hipocalcemia tras tiroidectomía total / Serum PTH levels as a predictive factor of hypocalcaemia after total thyroidectomy

Introducción: se analiza el valor de la determinación postoperatoria de paratirina como indicador de riesgo de hipocalcemia tras tiroidectomía total. Material y método: estudio prospectivo de 67 pacientes sometidos a tiroidectomía total por enfermedad benigna. Se determinó la concentración de paratirina y calcio iónico a las 20h de postoperatorio. Se calculó la sensibilidad, la especificidad y los valores predictivos positivo (VPP) y negativo (VPN) de las concentraciones de paratirina y calcio iónico para predecir la aparición de hipocalcemia sintomática o no. Resultados: presentaron hipocalcemia (Ca iónico<0,95 mmol/l) 42 pacientes (62,7%) pacientes, pero únicamente 20 (29,9%) mostraron síntomas. La concentración de paratirina a las 20h de la intervención fue inferior en el grupo con hipocalcemia sintomática (5,57±6,4pg/ml) que en el grupo de hipocalcemia sin síntomas (21,5±15,3pg/ml) y que entre los pacientes normocalcémicos (26,8±24,9pg/ml) (p=0,001). Con un punto de corte para la paratirina en 13pg/ml, la sensibilidad, la especificidad, el VPP y el VPN de paratirina para predecir la aparición de hipocalcemia analítica fueron del 54, el 72, el 76 y el 48%, respectivamente. Por otro lado, la sensibilidad para predecir hipocalcemia sintomática fue del 95% y la especificidad, el 76%. El test presentó alta incidencia de falsos positivos (11/30) (36%). El VPN fue del 97% y el VPP, el 65%. Paratirina y calcio iónico en el análisis multivariable fueron los únicos factores con valor predictivo como indicadores de riesgo de hipocalcemia sintomática. Conclusiones: una concentración normal de paratirina a las 20h de la intervención prácticamente descarta la aparición posterior de síntomas de hipocalcemia. Por contra, cifras de paratirina bajas no se acompañan necesariamente de síntomas debido al elevado número de falsos positivos (AU)

Introduction: Postoperative parathyroid hormone (PTH) levels as a predictor of hypocalcaemia in patients subjected to total thyroidectomy is analyzed. Matherial and method: Prospective study involving 67 patients who underwent total thyroidectomy due to a benign disease. Serum PTH and ionised calcium were measured 20h after surgery. Sensitivity, specificity and predictive values of PTH and ionised calcium levels were calculated to predict clinical and analytical hypocalcaemia. Results: A total of 42 (62.7%) patients developed hypocalcaemia (ionized calcium<0.95mmol/l), but only 20 (29.9%) presented with symptoms. PTH concentration the day after surgery was significantly lower in the group that developed symptomatic hypocalcaemia (5.57±6.4pg/ml) than in the asymptomatic (21.5±15.3pg/ml) or normocalcaemic (26.8±24.9pg/ml) groups (p=0.001). Taking the value of 13pg/ml as a cut-off point of PTH levels, sensitivity, specificity, positive predictive value and negative predictive value were 54%, 72%, 76% and 48%, respectively. On the other hand, sensitivity for predicting symptomatic hypocalcaemia was 95% and specificity was 76%. The test showed a high incidence of false positives (11/30, 36%). Negative predictive value was 97% and positive predictive value was 65%. In multivariate analysis, PTH and ionised calcium were the only perioperative factors that showed an independent predictive value as risk indicators of symptomatic hypocalcaemia. Conclusions: Normal PTH levels 20h after surgery practically rule out the subsequent appearance of hypocalcaemia symptoms. On the other hand, low PTH levels are not necessarily associated to symptomatic hypocalcaemia due to the high number of false positives (AU)