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OBJECTIVES: The aim of this study was to harmonize the criteria for the Bhattacharya indirect method Microsoft Excel Spreadsheet for reference intervals calculation to reduce between-user variability and use these criteria to calculate and evaluate reference intervals for eight analytes in two different years. METHODS: Anonymized laboratory test results from outpatients were extracted from January 1st 2018 to December 31st 2019. To assure data quality, we examined the monthly results from an external quality control program. Reference intervals were determined by the Bhattacharya method with the St Vincent's hospital Spreadsheet firstly using original criteria and then using additional harmonized criteria defined in this study. Consensus reference intervals using the additional harmonized criteria were calculated as the mean of four users' lower and upper reference interval results. To further test the operation criteria and robustness of the obtained reference intervals, an external user validated the Spreadsheet procedure. RESULTS: The extracted test results for all selected laboratory tests fulfilled the quality criteria and were included in the present study. Differences between users in calculated reference intervals were frequent when using the Spreadsheet. Therefore, additional criteria for the Spreadsheet were proposed and applied by independent users, such as: to set central bin as the mean of all the data, bin size as small as possible, at least three consecutive bins and a high proportion of bins within the curve. CONCLUSIONS: The proposed criteria contributed to the harmonization of reference interval calculation between users of the Bhattacharya indirect method Spreadsheet.
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Valores de Referencia , Humanos , Control de CalidadRESUMEN
Background and aims: GEM Premier ChemSTAT is a new point-of-care system providing rapid creatinine, BUN and tCO2 measurements together with electrolytes, metabolites, hematocrit, pH and pCO2 from a single whole blood specimen in acute care settings such as emergency departments and intensive care units. Accurate measurements of whole blood creatinine can aid in the diagnosis and treatment of renal diseases. Materials and methods: Heparinized whole blood samples from different clinical locations were evaluated on the GEM Premier ChemSTAT and results compared to plasma from the same samples on the Beckman AU5800 or whole blood on the GEM Premier 4000. Precision studies were conducted with whole blood and quality control material. Results: ChemSTAT correlated well with plasma samples on the AU5800 (regression slopes (S): 0.957-1.159, correlation coefficients (r)≥0.952) and with whole blood specimens on the GEM Premier 4000 (S: 0.9646-1.124, r ≥ 0.974). The repeatability was 0.1%-3.1% and QC precision were within lab and manufacturers' specifications. Conclusion: ChemSTAT demonstrated strong correlation to the comparative methods and excellent precision. Combining with its continuous quality management, ChemSTAT is suitable for acute care settings to provide rapid, reliable results, which could minimize time-to-treatment and improve patient outcome.
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The aim of this study was to determine reference intervals in an outpatient population from Vall d'Hebron laboratory using an indirect approach previously described in a Dutch population (NUMBER project). We used anonymized test results from individuals visiting general practitioners and analysed during 2018. Analytical quality was assured by EQA performance, daily average monitoring and by assessing longitudinal accuracy between 2018 and 2020 (using trueness verifiers from Dutch EQA). Per test, outliers by biochemically related tests were excluded, data were transformed to a normal distribution (if necessary) and means and standard deviations were calculated, stratified by age and sex. In addition, the reference limit estimator method was also used to calculate reference intervals using the same dataset. Finally, for standardized tests reference intervals obtained were compared with the published NUMBER results. Reference intervals were calculated using data from 509,408 clinical requests. For biochemical tests following a normal distribution, similar reference intervals were found between Vall d'Hebron and the Dutch study. For creatinine and urea, reference intervals increased with age in both populations. The upper limits of Gamma-glutamyl transferase were markedly higher in the Dutch study compared to Vall d'Hebron results. Creatine kinase and uric acid reference intervals were higher in both populations compared to conventional reference intervals. Medical test results following a normal distribution showed comparable and consistent reference intervals between studies. Therefore a simple indirect method is a feasible and cost-efficient approach for calculating reference intervals. Yet, for generating standardized calculated reference intervals that are traceable to higher order materials and methods, efforts should also focus on test standardization and bias assessment using commutable trueness verifiers.
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Laboratorios , Pacientes Ambulatorios , Creatina Quinasa , Creatinina , Humanos , Estándares de Referencia , Valores de ReferenciaRESUMEN
El río Ebro surca La Rioja dando lugar a una franja de tierra fértil de abundantes huertas y cultivos, muy especialmente el cultivo de la vid. Por otra parte, por sus tierras pasa la ruta jacobea (camino francés), fuente de transmisión de saber culinario. La historia documentada de la gastronomía riojanase remonta a la época medieval de la reconquista. Se mezclan la tendencia cristiana abundante de asados cárnicos, y la cocina algo más elaborada del Al-Ándalus. Gonzalo de Berceo, clérigo del Monasterio de Suso en San Millán de la Cogolla (La Rioja) menciona el vino y los quesos de la sierra de Cameros en el siglo XIII en sus versos. La cocina riojana está emparentada con la vasca, la navarra y la aragonesa. Se trata de una cocina en la que destaca la carne, las legumbres y el vino como sus más representativos componentes. También destaca por su abundante repostería, reflejo de una vida conventual extensa a lo largo del territorio. (AU)
The Ebro River flows through La Rioja giving rise to astrip of fertile land with abundant orchards and crops, especially vines. On the other hand, the Jacobean route (French way) passes through its lands, a source of transmission of culinary knowledge. The documented history of Rioja gastronomy dates back to the medieval period of the reconquest. The Christian trend plenty of meat roasts is mixed with the somewhat more elaborate cuisine of Al-Andalus. Gonzalo de Berceo, clergyman of the Monastery of Suso in San Millán de la Cogolla (La Rioja) mentions the wineand cheeses of the Sierra de Cameros in the 13th century in his verses. Riojan cuisine is related to Basque, Navarre and Aragonese cuisine. It is a cuisine in which meat, vegetables and wine stand out as its most representative components. It also stands out for its abundant confectionery, a reflection of extensive convent life throughout the territory. (AU)
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Humanos , Vino , Carne , Verduras , Culinaria , España/etnología , Libros de Cocina como AsuntoRESUMEN
Objectives: Administration of busulfan is extending rapidly as a part of a conditioning regimen in patients undergoing hematopoietic stem cell transplantation (HSCT). Monitoring blood plasma levels of busulfan is recommended for identifying the optimal dose in patients and for minimizing toxicity. The aim of this research was to validate a simple, rapid, and cost-effective analytical tool for measuring busulfan in human plasma that would be suitable for routine clinical use. This novel tool was based on liquid chromatography coupled to mass spectrometry. Methods: Human plasma samples were prepared using a one-step protein precipitation protocol. These samples were then resolved by isocratic elution in a C18 column. The mobile phase consisted 2 mM ammonium acetate and 0.1% formic acid dissolved in a 30:70 ratio of methanol/water. Busulfan-d8 was used as the internal standard. Results: The run time was optimized at 1.6 min. Standard curves were linear from 0.03 to 5 mg/L. The coefficient of variation (%CV) was less than 8%. The accuracy of this method had an acceptable bias that fell within 85-115% range. No interference between busulfan and the interfering compound hemoglobin, lipemia, or bilirubin not even at the highest concentrations of compound was tested. Neither carryover nor matrix effects were observed using this method. The area under the plasma drug concentration-time curves obtained for 15 pediatric patients who received busulfan therapy prior to HSCT were analyzed and correlated properly with the administered doses. Conclusions: This method was successfully validated and was found to be robust enough for therapeutic drug monitoring in a clinical setting.
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BACKGROUND: Different forms of pregenomic and other hepatitis B virus (HBV) RNA have been detected in patients' sera. These circulating HBV-RNAs may be useful for monitoring covalently closed circular DNA activity, and predicting hepatitis B e-antigen seroconversion or viral rebound after nucleos(t)ide analog cessation. Data on serum HBV-RNA quasispecies, however, is scarce. It is therefore important to develop methodologies to thoroughly analyze this quasispecies, ensuring the elimination of any residual HBV-DNA. Studying circulating HBV-RNA quasispecies may facilitate achieving functional cure of HBV infection. AIM: To establish a next-generation sequencing (NGS) methodology for analyzing serum HBV-RNA and comparing it with DNA quasispecies. METHODS: Thirteen untreated chronic hepatitis B patients, showing different HBV-genotypes and degrees of severity of liver disease were enrolled in the study and a serum sample with HBV-DNA > 5 Log10 IU/mL and HBV-RNA > 4 Log10 copies/mL was taken from each patient. HBV-RNA was treated with DNAse I to remove any residual DNA, and the region between nucleotides (nt) 1255-1611 was amplified using a 3-nested polymerase chain reaction protocol, and analyzed with NGS. Variability/conservation and complexity was compared between HBV-DNA and RNA quasispecies. RESULTS: No HBV-DNA contamination was detected in cDNA samples from HBV-RNA quasispecies. HBV quasispecies complexity showed heterogeneous behavior among patients. The Rare Haplotype Load at 1% was greater in DNA than in RNA quasispecies, with no statistically significant differences (P = 0.1641). Regarding conservation, information content was equal in RNA and DNA quasispecies in most nt positions [218/357 (61.06%)]. In 102 of the remaining 139 (73.38%), HBV-RNA showed slightly higher variability. Sliding window analysis identified 4 hyper-conserved sequence fragments in each quasispecies, 3 of them coincided between the 2 quasispecies: nts 1258-1286, 1545-1573 and 1575-1604. The 2 hyper-variable sequence fragments also coincided: nts 1311-1344 and 1461-1485. Sequences between nts 1519-1543 and 1559-1587 were only hyper-conserved in HBV-DNA and RNA, respectively. CONCLUSION: Our methodology allowed analyzing HBV-RNA quasispecies complexity and conservation without interference from HBV-DNA. Thanks to this, we have been able to compare both quasispecies in the present study.
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Ácidos Nucleicos Libres de Células , Hepatitis B Crónica , Hepatitis B , Antivirales/uso terapéutico , Estudios Transversales , ADN Viral/genética , ADN Viral/uso terapéutico , Hepatitis B/tratamiento farmacológico , Virus de la Hepatitis B/genética , Hepatitis B Crónica/diagnóstico , Hepatitis B Crónica/tratamiento farmacológico , Humanos , Cuasiespecies , ARNRESUMEN
Liver disease is frequently asymptomatic, challenging early identification in the primary care setting. The fibrosis 4 (FIB4) index is a liver fibrosis biomarker that is a potential alternative to liver biopsy for diagnosing and managing liver disease. This study aimed to calculate the FIB4 index for screening individuals at high risk of liver disease at the community level. This was a retrospective real-world study analyzing blood and serum test results from a central laboratory. The primary outcome was the number of individuals within each risk category for hepatic fibrosis: high risk (FIB4 ≥ 3.25) and low risk (FIB4 < 1.3). The analysis included samples from 31,753 patients, of which 18,102 were aged 40 to 75 years. In these patients, the FIB4 index had been explicitly requested in 1852 (10.2%) cases and estimated ad hoc in the rest. Of the 263 (1.5%) cases with FIB4 ≥ 3.25, the FIB4 index was requested in 46 (17.5%), and 52 (19.8%) showed evidence of liver fibrosis in their medical records, while the rest did not report any data regarding liver fibrosis. FIB4 is a simple score that can play a role as a "red flag" for early identification of patients at high risk of advanced liver fibrosis and their referral to specialized care.
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Objectives: The strain the SARS-COV-2 pandemic is putting on hospitals requires that predictive values are identified for a rapid triage and management of patients at a higher risk of developing severe COVID-19. We developed and validated a prognostic model of COVID-19 severity. Methods: A descriptive, comparative study of patients with positive vs. negative PCR-RT for SARS-COV-2 and of patients who developed moderate vs. severe COVID-19 was conducted. The model was built based on analytical and demographic data and comorbidities of patients seen in an Emergency Department with symptoms consistent with COVID-19. A logistic regression model was designed from data of the COVID-19-positive cohort. Results: The sample was composed of 410 COVID-positive patients (303 with moderate disease and 107 with severe disease) and 81 COVID-negative patients. The predictive variables identified included lactate dehydrogenase, C-reactive protein, total proteins, urea, and platelets. Internal calibration showed an area under the ROC curve (AUC) of 0.88 (CI 95%: 0.85-0.92), with a rate of correct classifications of 85.2% for a cut-off value of 0.5. External validation (100 patients) yielded an AUC of 0.79 (95% CI: 0.71-0.89), with a rate of correct classifications of 73%. Conclusions: The predictive model identifies patients at a higher risk of developing severe COVID-19 at Emergency Department, with a first blood test and common parameters used in a clinical laboratory. This model may be a valuable tool for clinical planning and decision-making.
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Coronavirus disease 2019 (COVID-19) is frequently associated with severe systemic consequences, including vasculitis, a hyperinflammatory state and hypercoagulation. The mechanisms leading to these life-threatening abnormalities are multifactorial. Based on the analysis of publicly available interactomes, we propose that severe acute respiratory syndrome coronavirus-2 infection directly causes a deficiency in C1 esterase inhibitor, a pathogen-specific mechanism that may help explain significant systemic abnormalities in patients with COVID-19.
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COVID-19/metabolismo , Proteína Inhibidora del Complemento C1/metabolismo , SARS-CoV-2/metabolismo , COVID-19/patología , HumanosRESUMEN
Aim: To assess agreement between fasting plasma glucose (FPG) and hemoglobin A1c (HbA1c) levels for diagnosis of dysglycemia (diabetes and risk of diabetes), overall and depending on clinical characteristics. Methods: The study enrolled 1020 adult subjects without drug-treated diabetes who underwent a laboratory test at a Spanish health care center. The criteria for dysglycemia of the American Diabetes Association were used. A logistic regression analysis was used to predict de novo diagnosis of dysglycemia based on sex, age, body mass index, anemia, and iron levels. Results: Overall prevalence of dysglycemia was 28.04%, and was identified by FPG only in 13.63% of subjects, by both FPG and HbA1c in 7.65%, and by HbA1c only in 6.76% (de novo diagnoses). Independent predictors of de novo diagnoses based on HbA1c were female sex (odds ratio [OR]: 2.119, 95% confidence interval [CI]: 1.133-4.020; p<0.020), age (OR for 42-56 years: 2.541, 95% CI: 0.634-17.140; OR for ≥57 years: 5.656, 95% CI: 1.516-36.980; overall p<0.007), and serum ferritin levels (borderline significance). Conclusions: In this study population, agreement between FPG and HbA1c for diagnosis of dysglycemia was poor, with FPG being the test that identified more subjects. De novo diagnoses based on HbA1c were more common in females and increased with age (AU)
Objetivo: Evaluar la concordancia entre el diagnóstico de disglucemia (diabetes y riesgo de diabetes) realizado por glucemia basal (GB) y hemoglobina A1c (HbA1c), globalmente y según características clínicas. Métodos: El estudio incluyó a 1.020 sujetos adultos con diabetes no tratada con fármacos que realizaron una prueba de laboratorio en un centro de salud español. Los criterios de disglucemia fueron los de la American Diabetes Association. Se utilizó un análisis de regresión logística para predecir un nuevo diagnóstico de disglucemia a partir del sexo, edad, índice de masa corporal, presencia de anemia y estatus férrico. Resultados: La prevalencia global de disglucemia fue del 28,04%, identificada únicamente por GB en el 13,63% de los sujetos, por GB y HbA1c en el 7,65% y solo por HbA1c en el 6,76% (nuevos diagnósticos). Los predictores independientes de nuevo diagnóstico según HbA1c fueron el sexo femenino (odds ratio [OR]: 2,119; intervalo de confianza [IC] 95%: 1,133-4,020; p<0,020), la edad (OR para 42-56 años: 2,541; IC 95%: 0,634-17,140; OR para ≥57 años: 5,656, IC 95%: 1,516-36,980; p<0,007 en general) y la ferritina sérica (significación límite). Conclusiones: En esta población la concordancia entre GB y HbA1c para el diagnóstico de disglucemia es pobre; la GB es la prueba que identifica más sujetos. Los nuevos diagnósticos por HbA1c se realizan con mayor frecuencia en mujeres y aumentan con la edad (AU)
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Humanos , Masculino , Femenino , Adulto , Glucemia , Diabetes Mellitus/diagnóstico , Hemoglobina Glucada/análisis , Sexo , Modelos Logísticos , Índice de Masa Corporal , Anemia/complicaciones , Anemia/diagnóstico , Intervalos de ConfianzaRESUMEN
AIM: To assess agreement between fasting plasma glucose (FPG) and hemoglobin A1c (HbA1c) levels for diagnosis of dysglycemia (diabetes and risk of diabetes), overall and depending on clinical characteristics. METHODS: The study enrolled 1020 adult subjects without drug-treated diabetes who underwent a laboratory test at a Spanish health care center. The criteria for dysglycemia of the American Diabetes Association were used. A logistic regression analysis was used to predict de novo diagnosis of dysglycemia based on sex, age, body mass index, anemia, and iron levels. RESULTS: Overall prevalence of dysglycemia was 28.04%, and was identified by FPG only in 13.63% of subjects, by both FPG and HbA1c in 7.65%, and by HbA1c only in 6.76% (de novo diagnoses). Independent predictors of de novo diagnoses based on HbA1c were female sex (odds ratio [OR]: 2.119, 95% confidence interval [CI]: 1.133-4.020; p<0.020), age (OR for 42-56 years: 2.541, 95% CI: 0.634-17.140; OR for ≥57 years: 5.656, 95% CI: 1.516-36.980; overall p<0.007), and serum ferritin levels (borderline significance). CONCLUSIONS: In this study population, agreement between FPG and HbA1c for diagnosis of dysglycemia was poor, with FPG being the test that identified more subjects. De novo diagnoses based on HbA1c were more common in females and increased with age.
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Factores de Edad , Glucemia/análisis , Diabetes Mellitus/sangre , Ayuno/sangre , Hemoglobina Glucada/análisis , Estado Prediabético/sangre , Factores Sexuales , Adulto , Anemia/sangre , Anemia/epidemiología , Índice de Masa Corporal , Comorbilidad , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/epidemiología , Femenino , Ferritinas/sangre , Humanos , Hierro/sangre , Sobrecarga de Hierro/sangre , Sobrecarga de Hierro/epidemiología , Masculino , Persona de Mediana Edad , Estado Prediabético/diagnóstico , Estado Prediabético/epidemiología , Prevalencia , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , España/epidemiologíaRESUMEN
BACKGROUND/AIMS: In myocytes from diabetic hearts, the reduction in the amplitude of the transient outward potassium current (I(to)) and the acceleration of its inactivation contribute to the action potential duration lengthening. Whereas the reduced amplitude is attributable to a reduced support of trophic factors, the mechanism underlying the acceleration of inactivation remains unknown. Ca(2+)/Calmodulin-dependent protein kinase II (CaMKII) modifies the inactivation kinetics of I(to). In this work we explored the role of CaMKII in the acceleration of I(to) current inactivation observed in diabetic myocytes. METHODS: We used patch-clamp and immunoblotting techniques in enzymatically-isolated myocytes from healthy and streptozotocin-induced diabetic rat hearts, and in blood samples from diabetic patients. RESULTS: In control myocytes, inhibition of either calmodulin or CaMKII accelerated I(to) current inactivation. However, in diabetic myocytes I(to) inactivation was already accelerated, and did not respond to calmodulin or CaMKII inhibition. Calmodulin protein abundance was significantly reduced in diabetic myocytes. Incubation of diabetic myocytes with insulin recovered calmodulin expression to normal values. A similar pattern of calmodulin expression appears in the blood of diabetic patients. Insulin treatment also restored I(to) current inactivation kinetics as well as the responsiveness to regulation by calmodulin. CONCLUSION: Diabetes-induced acceleration of I(to) current inactivation is due to a reduced effect of CaMKII on I(to) channels as a result of a diabetes-induced reduction in calmodulin protein expression. A correct follow up of the insulin treatment could prevent this alteration.