Performance and reference intervals of thrombin generation test: results from the Brazilian longitudinal study of adult health (ELSA-Brasil). A cross-sectional study

ABSTRACT BACKGROUND: The thrombin generation test (TGT) has shown promise for investigation of hemorrhagic and thrombotic diseases. However, despite its potential, it still needs standardization. Moreover, few studies have established reference values for TGT parameters. In Brazil, these values have not yet been established. OBJECTIVE: To determine TGT performance and reference intervals for TGT parameters in healthy individuals. DESIGN AND SETTING: Cross-sectional study conducted among participants in the Brazilian Longitudinal Study of Adult Health (Estudo Longitudinal de Saúde do Adulto, ELSA-Brasil). METHODS: The reference sample consisted of 620 healthy individuals. The calibrated automated thrombogram (CAT) method, under low and high tissue factor (TF) conditions, was used to assess thrombin generation. Test performance was analyzed using intra and interassay coefficients of variation (CV) and reference intervals were calculated using the nonparametric method proposed by the International Federation of Clinical Chemistry and the Clinical and Laboratory Standards Institute. RESULTS: The intraassay CV ranged from 1.4% to 2.2% and the interassay CV, 6.8% to 14.7%. The reference intervals for TGT parameters under low and high TF conditions were, respectively: lagtime: 3.0-10.3 and 1.4-3.7 min; endogenous thrombin potential (ETP): 1134.6-2517.9 and 1413.6-2658.0 nM.min; normalized ETP: 0.6-1.3 and 0.7-1.4; peak: 103.2-397.7 and 256.4-479.0 nM; normalized peak: 0.3-1.3 and 0.7-1.2; and time-to-peak: 5.6-16.0 and 3.4-6.7 min. These parameters were categorized relative to sex. Conclusion: TGT performance was adequate and the proposed reference intervals were similar to those of other studies. Our findings may be useful for consolidating the TGT, through contributing to its standardization and validation.

Electrical Cochlear Response as an Objective Measure of Hearing Threshold and Hearing Performance Evaluation in Pediatric Cochlear Implant Users / Respuesta Coclear Eléctrica como Medida Objetiva del Umbral Auditivo y la Evaluación del Rendimiento Auditivo en Usuarios Pediátricos de Implante Coclear

ABSTRACT The difficulties of applying the audiometry in pediatric populations and its methodological limitations in implanted patients have spurred the development of new alternative auditory evaluation methods. This study aimed to show an objective method to estimate hearing thresholds in pediatric cochlear implanted patients through Electrical Cochlear Response (ECR) and to quantify the hearing performance by using an Auditory Skills Questionnaire (ASQ) and a Calibrated Sounds Test (CST) designed on purpose. Eighteen implanted patients, 1-6 years old underwent standard audiometry, ECR, and ASQ in two evaluation sessions T1 and T2. At T2, in addition, patients underwent CST. For patients ≤3 years old (G1), Pure Tone Averages (PTA and PTAECR)showed a statistically significant difference between them at T1 and T2. At T2 improvements in audiometric and ECR thresholds were observed (p<0.05), regarding T1. Patients older than 3 years (G2) had significantly better ASQ and CST scores. CST detection scores at 40 dBHL for groups G1 and G2, 36% and 70% respectively, showed a better relationship to ECR thresholds. The relationship observed between ECR thresholds and CST detection scores seems to confirm that ECR brings the feasibility of objective hearing threshold estimation and provides a better frequency resolution than audiometry.

RESUMEN Las dificultades para la aplicación de la audiometría en la población pediátrica además de sus limitaciones metodológicas en pacientes usuarios de implante coclear, señalan la necesidad de métodos audiométricos alternos. En el presente trabajo se utiliza el potencial eléctrico, denominado Respuesta Coclear Eléctrica (ECR) observado solamente en usuarios de implante coclear, para la estimación de umbrales auditivos prescindiendo de la participación consiente del paciente, además de evaluar el desempeño auditivo mediante un Cuestionario de Habilidades Auditivas (ASQ) y la Prueba de Sonidos Calibrados (CST). A dieciocho participantes de 1 a 6 años, se les practicó Audiometría, ECR y ASQ en dos sesiones, T1 y T2; adicionalmente, en T2 se aplicó CST. En T1 y T2 los promedios de tonos puros, PTA y PTAECR, de pacientes ≤ 3 años (G1), mostraron una diferencia estadísticamente significativa entre ellos. En T2 los umbrales audiométricos y ECR (p <0.05), mejoraron respecto de T1. Pacientes > 3 años (G2) lograron puntuaciones ASQ y CST significativamente mejores. Los puntajes de detección CST a 40 dBHL, G1(36%) y G2(70%), mostraron mejor relación con los umbrales ECR. Esta relación entre los umbrales ECR y los puntajes de detección CST indican que la ECR permite estimar el umbral de audición, logrando adicionalmente mayor resolución en frecuencia que la audiometría.

Comparing the number of attempts required for complete nuclear chop using calibrated and conventional phacotip

Background: Phacoemulsification is a state of art technique with a steep learning curve the configuration of the phacotip affects the efficacy and execution of the nuclear chopping techniques. Inadequate penetration of phacotip may result in partial thickness nuclear cleavage with residual posterior plane and over enthusiastic penetration may result in posterior capsule rupture. This may be avoided if some estimate can be made preoperatively of the depth of penetration required to achieve full thickness crack.Methods: A total of 60 eyes of 60 patients with age related cataract with grade 4.0 to 6.9 (LOCS III) fulfilling the inclusion and exclusion criteria were included in the study after written informed consent. They were divided into two groups A and B with 30 patients each randomly using envelope method and were operated using conventional and calibrated phacotips respectively and the number of attempts required to achieve complete nuclear chop was noted.Results: In group A, vertical chop was safely and effectively done is 23 patients in just one attempt while it took 2 attempts in 6 patients and 3 attempts in 1 patient. In group B, it took just one attempt in 26 patients while 2 attempts were required in 4 patients. The two groups were comparable (p value 0.453) and was found to be statistically significant.Conclusions: With the use of a calibrated phacotip, the surgeon already knows to what depth he has to penetrate the tip into the center of nucleus depending upon the grade of cataract which results in safe and effective chop in minimum attempts where as in the conventional phacotip, it is more of a blind process. Hence the calibrated phacotip has taken the guess work out of question.

Accuracy of Blood Loss Determination after Vaginal Delivery: Visual Estimation versus Calibrated Measurement.

Background: The ability to determine accurately, the blood loss during childbirth is of extreme importance in the diagnosis and management of primary postpartum haemorrhage (PPH). Aims: In this study, we evaluate the effectiveness of visual estimation of blood loss, as a method of diagnosing primary postpartum haemorrhage. Methodology: A cross-sectional study on 271 participants was conducted between April and October 2012, at the Komfo Anokye Teaching Hospital. Women who went through spontaneous vaginal delivery were monitored and blood loss after delivery was visually estimated and then measured with a graduated receptacle, up to 1 hour post-delivery. The paired t-test and Bland- Altman plot was used to compare outcomes of the two methods. P<0.05 was considered significant. Results: Mean age of study participants was 27.7±5.7. As per visual estimation, most participants were reported to have lost <200 ml of blood (45.0%) as opposed to that of measured losses where most participants had lost between 200-500 ml of blood (48.7%). The mean measured blood loss was 306.09±218.39 ml against a 250±188.78 ml mean visually estimated blood loss (P < 0.0001). Prevalence of primary PPH as estimated with measured blood losses was 20.3% (55/271). Visual estimation, however gave a prevalence of 15.9% (43/271), an underestimation by 4.4%. A Bland- Altman plot showed a clinical bias large enough to cause significant differences in diagnosis of primary PPH. Conclusion: This study adds to existing evidence that the visual estimation of blood loss in clinical settings underestimates losses and is not reliable. This can lead to misdiagnosis of primary PPH and thus an underestimation of the condition. Health care workers should therefore base diagnosis on calibrated measurement methods.

Thrombin generation increasing with age and decreasing with use of heparin indicated by calibrated automated thrombogram conducted in Chinese / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Calibrated Automated Thrombogram(CAT) is a test to monitor the generation of thrombin. It can be described by four parameters: lag time, peak thrombin, endogenous thrombin potential (ETP) and time to peak (ttPeak). This study aims to determine the normal ranges of CAT parameters in Chinese, and evaluate whether thrombin generation is correlated with the concentration of heparin/low molecular weight heparin.</p><p><b>METHODS</b>Plasma from 120 healthy subjects were collected to determine the normal rangea of CAT parameters in Chinese. Normal plasma pool (NPP, n=25) spiked with different concentrations of heparin or enoxaparin were used to detecte CAT parameters. The overall and age specific normal ranges of CAT parameters were calculated using descriptive statistics method with mean±2SD. The correlation between CAT parameters and age or concentrations of heparin, enoxaparin were analyzed with linear regression model.</p><p><b>RESULTS</b>The normal ranges for lag time, peak thrombin, ETP, ttPeak in the subjects were 3.648±2.465 min, 367.39±151.93 nmol/L, 2277±1030 nmol/L•min and 6.372±4.280 min respectively. Age was linearly correlated with lag time (r=-0.6583, P<0.0001), peak thrombin (r=0.4863, P<0.0001), ETP (r=0.3608, P<0.0014) and ttPeak (r=-0.6313, P<0.0001). The values of ETP/peak ratio were linearly correlated with concentrations of heparin.</p><p><b>CONCLUSION</b>The normal ranges of four CAT parameters for Chinese were determined. CAT parameters are associated with age. ETP/peak ratio could be used to monitor the process of anticoagulation therapy.</p>

Asas metálicas calibradas para microbiólogos: Una alternativa de fabricación nacional / Calibrated metallic loops for microbiologists: A nationally manufactured alternative

Las asas calibradas para la cuantificación de unidades formadoras de colonias bacterianas transfieren un volumen de muestra “exactamente conocido” y han sido herramientas de gran importancia para laboratorios de control de calidad, investigación y clínico. Las asas que se comercializan en el país son importadas y mayoritariamente de aleaciones de níquel-cromo. Es por ello que surge la necesidad de buscar internamente alternativas de bajo costo. En este trabajo se propuso la fabricación de asas con alambres comerciales de níquel-cromo equivalentes a las asas de platino y níquel-cromo comerciales, a un costo inferior. Las asas fueron comparadas según su desempeño y apariencia. Todas las asas fueron calibradas empleando dos métodos (gravimétrico y colorimétrico) con ángulos de carga de 45º y 90°. Se empleó un diseño experimental con arreglo factorial de tratamiento 2³ (método de calibración-tipo de asa-ángulo de carga, con 4 repeticiones). Se realizaron análisis de varianza de 672 datos, comparación de medias (prueba de Tukey) y se calcularon los intervalos de confianza de Bonferroni de las desviaciones estándar. Se demostró que las asas artesanales emulan a las comerciales, representando una alternativa nacional de bajo costo para la obtención de estos instrumentos.

Calibrated metallic loops for the quantification of bacterial colony forming units transfer an “exactly known” sample volume and have been extremely important tools for quality control, research and clinical laboratories. The loops marketed in our country are imported and mostly made of nickel-chrome alloy. This has created the need for searching for low cost national alternatives. In this work we propose the manufacture of these loops with commercial nickel-chrome wire equivalent to the commercial platinum and nickel-chrome loops, at lower costs. The loops were comparable in performance and appearance. All the loops were calibrated using two methods (gravimetric and colorimetric) with 45º and 90º charge angles. An experimental design with factorial treatment arrangement 2³ (calibration method-loop type-charge angle with 4 repetitions) was used. A 672 data variance analysis and comparison of means (Tukey’s test) were carried out, and Bonferroni’s confidence intervals of the standard deviations were calculated. It was demonstrated that the artisan loops emulated the commercial ones, representing a low cost national alternative for obtaining these instruments.

Calibração da creatinina sérica com uso de equação de regressão: influência na performance da fórmula do estudo MDRD / Serum creatinine calibration with regression equation: influence on MDRD formula performance

Diferenças nos métodos de medida da creatinina sérica podem determinar amplas variações na taxa de filtração glomerular (TFG) estimada com fórmulas. Para a padronização da medida da creatinina, deve ser usado método calibrado rastreável para medida de referência com ID-MS (isotope dilution mass spectrometry). Objetivo: Avaliar a TFG estimada com a equação MDRD (Modification of Diet in Renal Disease) original (MDRDo, creatinina método não calibrado) e a equação MDRD re-expressa (MDRDr, método calibrado por ID-MS), comparando-as com a TFG medida pelo 51Cr-EDTA (método padrão) em indivíduos normais. Métodos: Foram avaliados 101 indivíduos, com idade média de 38±12 anos, sendo 45 homens. A TFG foi medida pela técnica de injeção única do 51Cr-EDTA (TFG 51Cr-EDTA) e estimada pelas equações MDRDo: 186 x creatinina sérica-1,154 x idade-0,203 x 0,742 (se mulher) x 1,210 (se negro) e MDRDr, substituindo-se o valor 186 por 175 na equação. A creatinina sérica foi medida pelo método de Jaffe não calibrado e transformada em calibrado com a fórmula: y=1,07x-0,249, obtida previamente por regressão. A concordância entre os métodos foi avaliada através da análise de Bland&Altman. Resultados: Os valores médios para as TFG 51Cr-EDTA, MDRDr e MDRDo foram de 105±18, 102±21 e 84±13 ml/min/1,73 m², respectivamente. Acurácia (percentual de casos de TFG estimada que não desviam em mais de 15% do valor medido) foi maior com o uso da MDRDr em relação à MDRDo (57% vs. 35%, P=0,002). O viés (diferença entre TFG medida e estimada) para 51Cr-EDTA e MDRDr foi de 3±23 ml/min/1,73 m². Para 51Cr-EDTA e MDRDo o viés foi significativamente maior, sendo de 21±18 ml/min/1,73 m². No entanto, a precisão, avaliada como desvio padrão do viés indicou elevada dispersão nos dois casos. Conclusão: O uso da equação re-expressa do MDRD, empregando a creatinina calibrada, produz uma estimativa mais acurada da TFG do que a equação original do MDRD.

Differences in methods of measurement of serum creatinine may provide wide variations in glomerular filtration rate (GFR) estimated with formulas. To standardize the measurement of creatinine, calibrated methods should be used, traceable to the reference ID-MS (isotope dilution mass spectrometry) method. Aim: To evaluate the performance of GFRs estimated with the original Modification of Diet in Renal Disease study equation (MDRDo; non-calibrated creatinine method) and with the re-expressed MDRD equation (MDRDr; ID-MS creatinine), comparing them with the GFR measured by 51Cr-EDTA (standard method ) in normal adults. Methods: 101 subjects, aged 38±12 years, 45 (45%) men were evaluated. GFR was measured by single-injection 51Cr-EDTA (GFR 51Cr-EDTA) technique and estimated by the following equations - MDRDo: 186 x serum creatinine-1. 154 x age-0.203 x 0.742 (if female) x 1.210 (if black), and MDRDr, replacing the value 186 by 175 in the equation. Serum creatinine was measured by a non-calibrated Jaffe’s method and transformed into calibrated with the formula: y=1.07x-0.249, previously obtained by regression. The agreement between methods was assessed by the Bland&Altman analyses. Results: The mean GFR 51Cr-EDTA, MDRDr and MDRDo were 105±18, 102±21 and 84±13 ml/min/1, 73 m2, respectively. There was no agreement between 51Cr-EDTA and MDRDo GFR (P <0.001), but it was present between 51Cr-EDTA and MDRDr GFR (P=0.149). Accuracy (percentage of cases of estimated GFR within 15% of measured value) was higher with the use of MDRDr in comparison to MDRDo (57% vs. 35%, P=0.002). Bias (diference between measured and estimated GFR) for 51Cr-EDTA and MDRDr was 3±23 ml/min/1.73 m². For 51Cr-EDTA and MDRDo the bias was significantly higher, 21±18 ml/min/1.73 m². However, the precision, evaluated as standard deviation of bias indicated a huge variation in both cases. Conclusion: The re-expressed MDRD equation, using calibrated creatinine, is a more accurate estimation of GFR than.