Evaluation of the Premier Hb9210 instrument for HbA1c determination.

Background: Glycated hemoglobin measurements are a valuable tool for long-term blood glucose monitoring and the diagnosis of diabetes. Its widespread use has been made possible due to the development of new analytical methods with improved performances and standardization with reference materials. The aim of the present study was to evaluate the Trinity Biotech Premier Hb9210 analyzer for the measurement of HbA1c.Methods: The precision was assessed using the CLSI EP-15A3 and EP-10A3 protocols. The latter was also used to investigate linearity, carryover, and linear drift. The comparison study was performed between Premier Hb910 and Tosoh HLC-723 G8 through Passing-Bablok regression and the Bland-Altman plot. The Fleiss Kappa index was used to assess the degree of agreement. The interference of Hb variants was investigated using samples with Hb variants S, C, D, E, J, and Seville.Results: Within-run and between-run imprecision fell between 0.37% and 1.16%. No statistically significant nonlinearity, carry-over, and/or drift were observed. The resulting regression line of the Passing-Bablok analysis was y = 0.00 + 1.00x. The Pearson correlation coefficient was 0.997. In the Bland-Altman plot, the relative bias was 0.01%. The overall Fleiss Kappa index was 0.9. No interference from hemoglobin variants was observed.Conclusion: The Premier Hb9210 demonstrated a high degree of automation, reproducibility, good agreement, minimal carry-over effect, and excellent linearity across the wide range of HbA1c levels commonly found in diabetic patients and was not influenced by Hb variants.

Premier Hb9210 can be used as an alternative to monitor glycemic status.Premier Hb9210 is not affected by common hemoglobin variants.Premier Hb9210 can correctly classify diabetic patients.

Liver stiffness accuracy by magnetic resonance elastography in histologically proven non-alcoholic fatty liver disease patients: a Spanish cohort.

OBJECTIVES: to evaluate the performance of magnetic resonance elastography (MRE) to stage liver fibrosis in patients with histologically confirmed nonalcoholic fatty liver disease (NAFLD) and to assess the impact of potential confounding factors in MRE diagnostic accuracy. The secondary objective was to compare MRE with other non-invasive methods for staging fibrosis such as transient elastography (TE) and non-invasive scores (APRI and FIB-4). METHODS: sixty-five histologically confirmed NAFLD patients were prospectively enrolled at the Hospital Universitario Virgen del Rocío (Seville, Spain). Liver stiffness was measured by MRE, TE and non-invasive scores (APRI and FIB-4). Fibrosis was assessed by liver biopsy using the steatosis, activity and fibrosis (SAF) score. Patients were classified into three groups according to the consistency between MRE and histopathological findings: underestimation, concordance and overestimation groups. Areas under the ROC curve (AUROC) and diagnostic performance were evaluated. RESULTS: the area under the ROC curve (AUROC) of MRE in advanced fibrosis (≥ F3) was 0.90 (0.82-0.97), while TE AUROC was 0.82 (0.72-0.93) (p = 0.22) and lower for the non-invasive test (FIB-4 0.67 and APRI 0.62). Inflammatory activity, steatosis grade and higher levels of liver biochemistry appeared to overestimate MRE results in the univariate analysis, but only gamma-glutamyl transferase (GGT) was statistically significant in the multivariate analysis (p < 0.01). Age, sex, body mass index (BMI), weight, diabetes mellitus (DM), high blood pressure (HBP), platelets or lipidic profile did not affect MRE accuracy. CONCLUSIONS: MRE is an effective and non-invasive method for detecting and staging liver fibrosis in NAFLD patients. MRE is more accurate than TE and allows the study of liver anatomy. Histological inflammation and surrogate biomarkers of inflammation can overestimate liver stiffness, but only GGT was statistically significant in the multivariate analysis. Important features of NAFLD patients such as obesity, DM, or lipidic profile did not affect MRE accuracy.

Liver stiffness accuracy by magnetic resonance elastography in histologically proven non-alcoholic fatty liver disease patients: a Spanish cohort

Objectives: to evaluate the performance of magnetic resonance elastography (MRE) to stage liver fibrosis in patients with histologically confirmed nonalcoholic fatty liver disease (NAFLD) and to assess the impact of potential confounding factors in MRE diagnostic accuracy. The secondary objective was to compare MRE with other non-invasive methods for staging fibrosis such as transient elastography (TE) and non-invasive scores (APRI and FIB-4). Methods: sixty-five histologically confirmed NAFLD patients were prospectively enrolled at the Hospital Universitario Virgen del Rocío (Seville, Spain). Liver stiffness was measured by MRE, TE and non-invasive scores (APRI and FIB-4). Fibrosis was assessed by liver biopsy using the steatosis, activity and fibrosis (SAF) score. Patients were classified into three groups according to the consistency between MRE and histopathological findings: underestimation, concordance and overestimation groups. Areas under the ROC curve (AUROC) and diagnostic performance were evaluated. Results: the area under the ROC curve (AUROC) of MRE in advanced fibrosis (≥ F3) was 0.90 (0.82-0.97), while TE AUROC was 0.82 (0.72-0.93) (p = 0.22) and lower for the non-invasive test (FIB-4 0.67 and APRI 0.62). Inflammatory activity, steatosis grade and higher levels of liver biochemistry appeared to overestimate MRE results in the univariate analysis, but only gamma-glutamyl transferase (GGT) was statistically significant in the multivariate analysis (p < 0.01). Age, sex, body mass index (BMI), weight, diabetes mellitus (DM), high blood pressure (HBP), platelets or lipidic profile did not affect MRE accuracy. Conclusions: MRE is an effective and non-invasive method for detecting and staging liver fibrosis in NAFLD patients. MRE is more accurate than TE and allows the study of liver anatomy (AU)

Hypoparathyroidism after thyroidectomy: A 20-year experience at a children's hospital.

INTRODUCTION: Hypoparathyroidism (HP) is the most common complication of total thyroidectomy and can be an emergency. OBJECTIVES: To describe the prevalence of HP after total thyroidectomy in children under 14 years of age, the variables related to its appearance and its clinical expression. PATIENTS AND METHODS: Retrospective study at a children's hospital in the last 20 years. HP was defined by the need to supplement calcium after the intervention and was considered permanent if it could not be suspended within 12 months. Fisher's statistical method of comparison of proportions. RESULTS: Thirty-nine children and adolescents (26 females) with an age range of 3.67-14.00 years. In 25 patients, the intervention was prophylactic and in 14 it was therapeutic; 14 suffered accidental excision of some parathyroid gland, but none more than two of them; 12 presented HP, of which 3 were permanent; 5 presented clinical symptoms; 1 of them was an emergency. The frequency of HP was 4/4 when 2 parathyroids were dissected, 2/10 when one was dissected, and 6/25 when none were dissected (p = 0.02). In the prophylactic interventions, it was 6/25 compared to 6/14 in the therapeutic ones (p = 0.29). The three cases of permanent HP were in children under 6 years of age, and it did not occur in any older children (p = 0.09). CONCLUSIONS: HP is a common and sometimes serious complication in children after total thyroidectomy. It can occur, and even be permanent, even if the intervention is prophylactic and parathyroid glands remain in situ. Younger age could be a risk factor.

Hypoparathyroidism after thyroidectomy: A 20-year experience at a children's hospital. / Hipoparatiroidismo postiroidectomía: experiencia de 20 años en un hospital infantil.

INTRODUCTION: Hypoparathyroidism (HP) is the most common complication of total thyroidectomy and can be an emergency. OBJECTIVES: To describe the prevalence of HP after total thyroidectomy in children under 14 years of age, the variables related to its appearance and its clinical expression. PATIENTS AND METHODS: Retrospective study at a children's hospital in the last 20 years. HP was defined by the need to supplement calcium after the intervention and was considered permanent if it could not be suspended within 12 months. Fisher's statistical method of comparison of proportions. RESULTS: Thirty-nine children and adolescents (26 females) with an age range of 3.67 to 14.00 years. In 25 patients, the intervention was prophylactic and in 14 it was therapeutic. Fourteen suffered accidental excision of some parathyroid gland, but none more than t2 of them. Twelve presented HP, of which 3 were permanent; 5 presented clinical symptoms; one of them was an emergency. The frequency of HP was 4/4 when 2 parathyroids were dissected, 2/10 when one was dissected, and 6/25 when none were dissected (P=.02). In the prophylactic interventions, it was 6/25 compared to 6/14 in the therapeutic ones (P=.29). The 3 cases of permanent HP were in children under 6 years of age, and it did not occur in any older children (P=.09). CONCLUSIONS: HP is a common and sometimes serious complication in children after total thyroidectomy. It can occur, and even be permanent, even if the intervention is prophylactic and parathyroid glands remain in situ. Younger age could be a risk factor.

Gestante eutiroidea con anticuerpos estimulantes del receptor de tirotropina positivos: manifestación tardía de hipertiroidismo neonatal. A propósito de un caso / Euthyroid pregnant woman with positive thyrotropin receptor-stimulating antibodies: Late manifestation of neonatal hyperthyroidism. About a case

Las enfermedades tiroideas, después de la diabetes mellitus, se encuentran entre los trastornos endocrinos más comunes durante el embarazo, con una incidencia del 5-10%. Es importante su detección y tratamiento precoz ya que puede tener consecuencias negativas tanto para la madre como para el feto. El hipertiroidismo se encuentra en menor frecuencia que el hipotiroidismo durante el embarazo, entre 0,1-1%. Se caracteriza por presentar tirotropina baja con hormonas tiroideas elevadas, siendo la enfermedad de Graves la causa más frecuente (el 85% de los casos). A continuación se expone el caso de un lactante con hipertiroidismo primario de etiología autoinmune, hijo de una madre sin diagnóstico previo de hipertiroidismo durante la gestación (AU)

Thyroid diseases, after diabetes mellitus, are among the most common endocrine disorders during pregnancy, with an incidence of 5-10%. Early detection and treatment is important, as they can have negative consequences for both the mother and the foetus. Hyperthyroidism is less frequent than hypothyroidism during pregnancy, being between 0.1% and 1%. It is characterised by a low thyrotropin with elevated thyroid hormones, with Graves’ disease being the most frequent cause (85% of cases). The following is the case of an infant with primary hyperthyroidism of autoimmune origin, the son of a mother without previous diagnosis of hyperthyroidism during gestation (AU)

Monitoring of transplanted liver health by quantification of organ-specific genomic marker in circulating DNA from receptor.

BACKGROUND: Health assessment of the transplanted organ is very important due to the relationship of long-term survival of organ transplant recipients and health organ maintenance. Nowadays, the measurement of cell-free DNA from grafts in the circulation of transplant recipients has been considered a potential biomarker of organ rejection or transplant associated complications in an attempt to replace or reduce liver biopsy. However, methods developed to date are expensive and extremely time-consuming. Our approach was to measure the SRY gene, as a male organ biomarker, in a setting of sex-mismatched female recipients of male donor organs. METHODS: Cell-free DNA quantization of the SRY gene was performed by real-time quantitative PCR beforehand, at the moment of transplantation during reperfusion (day 0) and during the stay at the intensive care unit. Beta-globin cell-free DNA levels, a general cellular damage marker, were also quantified. RESULTS: Beta-globin mean values of patients, who accepted the graft without any complications during the first week after surgery, diminished from day 0 until patient stabilization. This decrease was not so evident in patients who suffered some kind of post-transplantation complications. All patients showed an increase in SRY levels at day 0, which decreased during hospitalization. Different complications that did not compromise donated organs showed increased beta-globin levels but no SRY gene levels. However, when a donated organ was damaged the patients exhibited high levels of both genes. CONCLUSION: Determination of a SRY gene in a female recipient's serum is a clear and specific biomarker of donated organs and may give us important information about graft health in a short period of time by a non-expensive technique. This approach may permit clinicians to maintain a close follow up of the transplanted patient.

Intraoperative parathyroid hormone monitoring in neonatal severe primary hyperparathyroidism.

Neonatal severe primary hyperparathyroidism presents in the first days of life with severe life-threatening hypercalcemia. It is associated with an inactivating homozygous mutation of the calcium sensing receptor gene. Total parathyroidectomy is the treatment of choice, so the surgeon must identify all the parathyroid tissue, including supernumerary and ectopic glands. We present the case of an infant who underwent total parathyroidectomy at age 4 months in which intraoperative parathyroid hormone monitoring provided immediate confirmation of surgical cure.

Comparación entre 2 sistemas analíticos para la determinación de la hemoglobina A1c: inmunoturbidimetría versus cromatografía líquida de alta eficiencia / Comparison of 2 analytical systems for hemoglobin A1c: An immunoturbidimetric method versus high performance liquid chromatography

Introducción. La hemoglobina A1c (HbA1c) es ampliamente utilizada en la determinación del estado glucémico de pacientes con diabetes mellitus. El objetivo de este estudio fue comparar 2 métodos automatizados para medir HbA1c basados en diferentes principios de medida, evaluar la correlación entre ambos y su practicabilidad. Métodos. Se analizaron 622 muestras mediante 2 sistemas analíticos con fundamentos diferentes de medición: cromatografía líquida de alta eficiencia (HPLC) (analizador ADAMS A1c HA-8160; A. Menarini Diagnostics, Italia) e inmunoturbidimetría (Tina-quant Hemoglobin A1c Gen.3, plataforma Cobas 6000; Roche Diagnostics, Suiza). Ambos métodos fueron calibrados según el procedimiento de referencia de la IFCC. Se valoró la correlación entre ambos métodos mediante los análisis de regresión de mínimos cuadrados y Passing-Bablok (R programa v.2.11.1). También se registró el tiempo de puesta en marcha, las tareas de mantenimiento diario y el rendimiento de los 2 instrumentos. Resultados. La correlación fue muy alta tanto por mínimos cuadrados (ordenada en el origen 0.05, pendiente 0.98) como en Passing-Bablok (ordenada en el origen 0,10, pendiente 1,00). El tiempo invertido diariamente para la puesta en marcha del analizador HA-8160 fue de 25 min y el tiempo de finalización fue de 15 min. Las tareas de mantenimiento del Cobas 6000 al inicio y fin del día son procesos automatizados. El rendimiento de los analizadores fue 20 muestras/h en el HA-8160 y 100 muestras/h en el Cobas 6000. El sistema analítico cuyo principio de medida es HPLC incluye también el análisis manual de cada cromatograma. Conclusiones. Existe una correlación excelente entre los métodos de HPLC e inmunoturbidimétrico. La ventaja del sistema analítico que utiliza la inmunoturbidimetría es la optimización del tiempo de procesamiento de las determinaciones de HbA1c, lo que reduce el coste unitario de la prueba (AU)

Introduction. Hemoglobin A1c (HbA1c) is widely used to assess glycemic status in patients with diabetes mellitus. The purpose of this study was to compare 2 automated analytical systems to measure HbA1c that use different measurement principles, evaluating the correlation between the two methods, as well as their ease of use. Methods. A total of 622 samples were analyzed using 2 methods: high performance liquid chromatography (HPLC) (analyzer ADAMS A1c HA-8160; A. Menarini Diagnostics, Italy) and an immunoturbidimetric assay (Tina-quant Hemoglobin A1c Gen.3, Cobas 6000 analyzer; Roche Diagnostics, Switzerland). Both methods were calibrated in accordance with IFCC reference measurement procedure. The correlation between the two methods was assessed by least squares and Passing-Bablok linear regression analyses (R program v.2.11.1). The daily start-up time of the 2 instruments used, daily maintenance tasks, and determination of throughput were also recorded. Results. There was a strong correlation between the results generated by the two test methods using both the least squares (intercept 0.54; slope 0.98) and Passing-Bablok (intercept 0.10; slope 1.00) regression methods. The time spent daily for the start-up of the HA-8160 analyzer was 25 min and completion time was 15 min. Maintenance tasks for the Cobas 6000 analyzer at the beginning and end of the day are automated processes. The throughput for the HA-8160 analyzer was 20 samples/h, and 100 samples/h for the Cobas 6000 analyzer. The HPLC method also included a time-consuming manual analysis of each chromatogram. Conclusions. An excellent correlation was observed between the HPLC and immunoturbidimetric methods. The advantages of the immunoturbidimetric method are optimization of processing time of HbA1c tests and a reduction in the unit cost per test (AU)

Estudio de autoanticuerpos en el inicio de la diabetes autoinmune en nuestro medio mediante ELISA / Autoantibodies: analysis of new-onset autoimmune diabetes using ELISA test

Introducción. Los auto-anticuerpos son marcadores útiles en el inicio de la diabetes mellitus autoinmune, como apoyo diagnóstico. El objetivo de este trabajo fue conocer la prevalencia de autoanticuerpos en el inicio de diabetes en nuestro medio, medidos mediante ELISA y valorar su papel diagnóstico en diabetes autoinmune. Material y métodos. Muestra de 111 pacientes con diabetes de inicio: 61 tipo 1 (incluye 12 diabetes autoinmune latente del adulto [LADA]) y 50 tipo 2. Grupo control 64 no diabéticos. Se analizaron antidescarboxilasa del ácido glutámico GADA, anti-tirosina fosfatasa de membrana IA-2 y anti-insulina IAA, mediante enzimoinmunoanálisis ELISA en microplaca. Se realizó estudio observacional descriptivo para valoración de pruebas diagnósticas. Programa estadístico PASW Statistics versión 18. Resultados. GADA(+): 78,7% DM1; 83,3% LADA; 2,0% DM2; 1,6% control. IA-2(+): 52,2% DM1; 45,5% LADA; 16,4% DM2; 12,5% control. IAA(+): 10,3% DM1; 18% LADA; 6,0% DM2; 1,7% control. GADA e IA-2 diferenciaron significativamente (p<0,0001) a los pacientes con diabetes autoinmune. No así IAA. Área bajo curva receiver operating characteristics (ROC): GADA=0,90 (p<0,0001); IA-2=0,74 (p<0,0001); IAA=0,57 (p=0,126). Criterio límite GADA(+) > 5,5 U/mL (sensibilidad 79%, especificidad 98%) e IA-2(+) > 6,0 U/mL (sensibilidad 54%, especificidad 84%). Análisis de factores riesgo asociados: Odds ratio GADA=51,44 e IA-2=4,64. Conclusiones. En nuestro estudio, la prevalencia de GADA en el inicio de diabetes es alta y su determinación eficaz en el diagnóstico de diabetes autoinmune. IA-2 incrementó 4,6 veces la probabilidad diagnóstica. IAA medidos mediante nuestro test ELISA no han mostrado valor como marcadores en la diabetes autoinmune, aunque esto no descarta su utilidad en otras patologías (AU)

Autoantibodies are useful markers for clinical onset of autoimmune diabetes, and as a diagnostic supports. The aim of this study was to determine the prevalence of autoantibodies at the onset diabetes mellitus in our environment, measured by an ELISA technique, and to assess its role as autoimmune diabetes diagnostic support. Material and methods. A sample of 111 patients with diabetes onset was studied, which included, 61 type 1 diabetes (12 Latent Autoimmune Diabetes in Adults (LADA) and 50 type 2 diabetes. Control group: 64 non-diabetics. Antibodies glutamic acid decarboxylase antibodies (GADA), membrane phosphatase anti-tyrosine (IA-2), and anti-insulin (IAA) were determined by enzyme immunoassay ELISA microplates. A descriptive observational study was conducted to the measure diagnostic tests using the software PASW Statistics version 18. Results. GADA(+): 78.7% DM1; 2.0% DM2; 1.6% control. IA-2(+): 52.2% DM1; 45.5% LADA; 16.4% DM2; 12.5% control. IAA(+): 10.3% DM1; 18.0% LADA; 6.0% DM2; 1.7% control. Comparing means, autoimmune diabetics were significantly differentiated (P<.0001) by GADA and IA-2 values, but not with IAA. The area under the Curve using software Receiver Operating Characteristics (ROC): GADA=0.90 (P<.0001), IA-2=0.74 (P<.0001), IAA=0.57 (P=.126). Optimal cut off for GADA>5.5U/mL (sensitivity 79%, specificity 98%) and IA-2>6.0U/mL (sensitivity 54%, specificity 84%). On analysing risk factors associated with autoimmune diabetes diagnosis, the calculated Odds ratio gave GADA=51.44 and IA-2=4.64. Conclusions. In this study GADA(+) prevalence is high at diabetes onset, and its calculation has been effective for autoimmune diabetes diagnosis. IA-2 increases diagnostic probability 4.6 times. Anti-insulin antibodies measured by ELISA test did not demonstrate value for autoimmune diabetes diagnosis and classification, although its usefulness is not excluded for the diagnosis support of other diseases (AU)

Hemoglobin Seville [α2 ß2 81(EF5) Leu→Phe] a silent phenotypic variant that interferes in hemoglobin A1c measurement by ion-exchange HPLC method.

OBJECTIVES: The aim of the study was to investigate hemoglobin (Hb) species in a 61 year-old male with diabetes mellitus type II and a low value of Hb A(1c). DESIGN AND METHODS: Hb species were analyzed by electrophoresis and chromatography methods. Functional properties were determined by oxygen equilibrium studies. ß-globin gene was amplified by PCR and sequenced. RESULTS AND CONCLUSIONS: A novel clinically silent Hb (Hb Seville), that results in falsely low Hb A(1c) measurement, was detected. This Hb variant presented a single base mutation at codon 81 (C→T) of the ß-globin gene. This case points out the necessity of careful inspection of the chromatograms and the use of additional methods to Hb A(1c) measurement when the presence of aberrant peaks is detected.

Influence of reduction in the elution times on HPLC glycohaemoglobin results.

OBJECTIVES: To compare HPLC methods with short and long elution times for HbA1c measurement in blood. METHODS: Comparison between G7-Tosoh (1.2 min); Bio-Rad-Variant-II-Turbo (1.3 min) and Arkray-HA-8160 (2.9 min). RESULTS: Passing-Bablok regression equations were: Y=0.17+0.96X; Y=-0.39+1.01X; Y=-0.40+1.0X and the means of the differences using Bland-Altman Plot were 0.02; -0.34; 0.32 for HA-8160/G7-Tosoh, HA-8160/Variant-II-Turbo and G7-Tosoh/Variant-II-Turbo, respectively. CONCLUSIONS: Faster elution methods had no problems on reproducibility of results obtained by slower elution methods.

Características citomorfológicas e inmunohistoquímicas de cultivos primarios obtenidos a partir de tumores cerebrales / Cyto-morphological and immunohistochemical features of primary cell cultures obtained from human brain tumor

Objetivos: Establecer si los cultivos primarios de tumores cerebrales son válidos como método de estudio 'in vitro' para la extrapolación y aplicación clínica futura de ensayos terapéuticos. Material y método: Se obtuvieron 8 cultivos primarios a partir del material de biopsia de pacientes con tumores cerebrales (2 glioblastomas multiformes, 1 ependimoma, 1 oligodendro-glioma anaplásico y:4 meningiomas). Se valoró la expresión inmunohistoquímica a la proteína ácida glial fibrilar, vimentina, antígeno epitelial de membrana, S-100 y CD57 en el material turnoral parafinado y -en los cultivos primarios correspondientes. Resultados: Se obtuvo una estrecha correlación en la expresión inmunohistoquímica, para cada tumor a los antígenos estudiados, entre el material parafinado y los cultivos primarios (23/24, 95.8 por ciento). únicamente se apreció una disminución en la intensidad de dicha expresión, cuando esta era positiva en el material parafinado.. (7/18, 38.9 por ciento), en el cultivo primario correspondiente, pero en ningún caso con resultados negativos en parafina se evidenció positividad en el cultivo celular (0/6, 0 por ciento). Conclusiones: Las técnicas inmunohistoquímicas son válidas para caracterizar a los elementos celulares de los cultivos primarios obtenidos de tumores cerebrales, permitiéndonos complementar a otros métodos como la citomorfología y validarlos como 'banco de pruebas' para futuros estudios con aplicación clínica y terapéutica (AU)