RESUMO
ObjectiveTo investigate the clinical appropriateness and application value of the peroxidase （POD） method for the detection of unbound bilirubin （UB） in neonatal serum. MethodsHydrogen peroxide （0.33 mol/L） and three different final concentrations （0.019， 0.038， 0.075 μg/mL） of horseradish peroxidase （HRP） were added to standard bilirubin solution （1， 2， 3 μmol/L） to obtain a standardized HRP primary rate constant Kp. Then 25 μL of neonatal serum was diluted by 41.6 fold， and measured with 2.4 and 4.8 μg/mL HRP at 37 ℃ under the dark， to determine the UB concentration. The accuracy， precision， and stability of the methodology were validated. The clinical characteristics of 33 jaundiced neonates were collected， including total serum bilirubin （TSB）， indirect bilirubin （IDB）， albumin （ALB）， bilirubin to albumin molar ratio （BAMR）， etc. The experimental data were analyzed by Graphpad Prism 8.0. ResultsA standardized Kp of （7.20±1.08） mL·μg-1·min-1 was determined at pH 7.4±0.2， 37 ℃ in the dark. The HRP activity and UB concentrations remained stable at -20 ℃ for 3 weeks and a week， respectively. The mean intra-day and inter-day coefficients of variation of the serum samples with different UB concentrations were less than 10%. In this study， the UB concentrations in 33 jaundiced neonates （gestational age ≥35 weeks） were measured by the POD method in the range of （0.32～1.20） μg/dL， which was positively correlated with TSB， IDB and BAMR. Of the five infants whose UB concentrations measured more than 1 μg/dL， three received intensive phototherapy （60%）. ConclusionsThe POD method combined with a standard equipment spectrophotometer to detect serum UB concentrations in neonates is easy to operate， rapid to detect， and low cost. This method has good accuracy and precision， which is convenient for clinical implementation. Moreover， the measurement of serum UB may assist us in better management of neonatal jaundice in clinical practice.
RESUMO
Comparar la evaluación de morfología espermática mediante dos tinciones: Testsimplets® y Diff-Quik y la eficiencia de Testsimplets® y el test de peroxidasa para evaluar leucocitos en semen. Comparamos la morfología espermática de las muestras seminales de 30 pacientes infértiles, evaluada a través de las tinciones Testsimplets® y Diff-Quik. Adicionalmente, en pacientes que presentaron más de 106 de células redondas por mL de semen (n=7), comparamos la evaluación de leucocitos por Testsimplets® y el test de peroxidasa. En el Centro de fertilidad UNIFERTES, en Caracas, Venezuela. No existen diferencias significativas en el porcentaje de espermatozoides morfológicamente normales, siguiendo el criterio estricto de Kruger, entre ambas tinciones. Encontramos diferencias significativas en los porcentajes de los siguientes defectos morfológicos espermáticos, entre tinción: cabeza piriforme, pieza media doblada, irregular o con gota citoplasmática. No se encontraron diferencias significativas en la evaluación de leucocitos en semen mediante Testsimplets® y el test de peroxidasa. Testsimplets® es práctico para ahorrar tiempo y, consecuentemente, mejorar la eficiencia en el tiempo de entrega de resultados. Sin embargo, es considerablemente más costoso que Diff-Quik
To compare sperm morphology by Testsimplets® and Diff-Quik staining. To compare the efficiency of Staining cellular peroxidase and Testsimplets® to assess leukocytes in semen. We compared sperm morphology of 30 infertile patients evaluated by Testsimplets® and Diff-Quik staining. Additionally, in patients with more than 106 round cells per ml semen (n=7), we compared the assessment of leukocytes by Staining cellular peroxidase and Testsimplets®. Fertility clinic UNIFERTES, in Caracas, Venezuela. There was no significant difference in the percentage of morphologically normal sperm, according to Krugers strict criteria, between both stains. We found significant differences in the percentages of the following sperm morphological defects between staining: pyriform head, folded middle piece, irregular and presence of cytoplasmic droplets. There was no significant difference in the assessment of leukocytes in semen by Staining cellular peroxidase and Testsimplets®. Testsimplets® is practical for saving time and consequently, improving the efficiency in the delivery time of results. Nonetheless, it is considerably more expensive than Diff-Quik
