The impact of therapeutic hypothermia on peripheral blood cell in newborns with hypoxic ischemic encephalopathy

Abstract The effect of hypothermia treatment on white blood cell (WBC), neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR) and platelet-to-lymphocyte ratio (PLR) values as an indicator of inflammation was evaluated in newborns with hypoxic ischemic encephalopathy (HIE). The study was performed that the before-therapeutic hypothermia (TH) and after-TH WBC, lymphocytes, neutrophils, monocytes and NLR, LMR and PLR values of the complete blood cell count were retrospectively evaluated. The results of the patient group were compared with the results of healthy newborns. A total of 78 patients who underwent TH were evaluated in our study. Mean values before and after TH were NLR3.8/2.7, LMR 5.6/8.6, and PLR 60.3/67.1 respectively. A statistical significance was present for NLR values before and after TH in those with seizure in our study (4.15±2.95/3.01±2.54) but no statistical significance was found for LMR or PLR. In neonates with HIE, effect of TH on complete blood cell count and inflammatory mechanisms (mediated neutrophil and lymphocyte) may be minimal.

Diagnosis of patients with blood cell count for COVID-19: An explainable artificial intelligence approach / Diagnóstico de pacientes com hemograma para COVID-19: Uma abordagem com inteligência artificial explicável / Diagnóstico de pacientes con hemograma para COVID-19: un enfoque explicable de inteligencia artificial

Objective: Present an explainable artificial intelligence (AI) approach for COVID-19 diagnosis with blood cell count. Methods: Five AI algorithms were evaluated: Logistic Regression, Random Forest, Support Vector Machine, Gradient Boosting and eXtreme Gradient Boosting. A Bayesian optimization with 5-Fold cross-validation was used to hyper-parameters tuning. The model selection evaluated three results: cross validation performance, test set prediction performance and a backtest: performance on identifying patients negative for COVID-19, but positive for others respiratory pathologies. Shapley Additive explanations (SHAP) was used to explain the chosen model. Results: A Random Forest model was obtained with 77.7% F1-Score (IC95%:57.1;92.3), 85.9% AUC (IC95%:73.7;95.9), 74.4% Sensitivity (IC95%:50.0;92.1) and 97.5% Specificity (IC95%:93.6;100.0). The main features were leukocytes, platelets and eosinophils. Conclusion: The research highlights the importance of model interpretability, demonstrating blood cell count as a possibility for COVID-19 diagnosis. The methodological structure developed, using TRIPOD's guidelines, can be extrapolated to other pathologies.

Objetivo: Propor uma abordagem com inteligência artificial explicável para diagnóstico de COVID-19 com hemograma. Métodos: Cinco algoritmos de IA foram testados: Regressão Logística, Florestas Aleatórias, Máquina de Vetores de Suporte, Gradient Boosting e eXtreme Gradient Boosting. Os hiper-parâmetros foram definidos através da otimização bayesiana com validação cruzada 5-Fold. A seleção de modelo utilizou três resultados de desempenho para definir o melhor modelo: validação cruzada, conjunto de teste e rendimento na identificação de pacientes negativos para COVID-19, porém positivos para outras patologias respiratórias (backtest). Ao final, Shapley Additive explanations (SHAP) foi utilizado para explicar o modelo escolhido. Resultados: Obteve-se um modelo Random Forest com F1-Score de 77.7% (IC95%:57.1;92.3), AUC de 85.9% (IC95%:73.7;95.9), Sensibilidade de 74.4% (IC95%:50.0;92.1) e Especificidade de 97.5% (IC95%:93.6;100.0). As principais variáveis foram leucócitos, plaquetas e eosinófilos. Conclusão: A pesquisa destaca a importância da interpretabilidade do modelo, demonstrando o hemograma como uma possibilidade para diagnosticar COVID-19. A estrutura metodológica desenvolvida no estudo, utilizando as diretrizes do TRIPOD, pode ser extrapolada para detecção de outras patologias.

Objetivo: Proponer un enfoque explicable de inteligencia artificial (IA) para el diagnóstico de COVID-19 con el uso de hemograma. Métodos: Cinco modelos de IA fueron evaluados: Logistic Regression, Random Forest, Support Vector Machine, Gradient Boosting e eXtreme Gradient Boosting. Los hiper-parámetros fueron definidos a través de optimización bayesiana con validación cruzada 5-Folds. La selección del modelo se utilizó tres resultados: rendimiento del validación cruzada, rendimento en conjunto de pruebas y el análisis de desempeño en identificación de pacientes negativos para COVID-19, pero positivos para otras patologías respiratorias (backtest). Shapley Additive explanations (SHAP) fue utilizado para explicar el modelo elegido. Resultados: Se obtuvo un modelo Random Forest con F1-Score de 77.7% (IC95%:57.1;92.3), AUC de 85.9% (IC95%:73.7;95.9), Sensibilidad de 74.4% (IC95%:50.0;92.1) y Especificidad de 97.5% (IC95%:93.6;100.0). Las principales variables fueron leucocitos, plaquetas y eosinófilos. Conclusión: La investigación presenta la importancia de la interpretabilidad del modelo, demostrando el uso de hemograma como posibilidad para diagnosticar COVID-19. La estructura elaborada, siguiendo las directrices de TRIPOD, puede ser extrapolar para otras patologías.

Comparative study on erythrogram parameters of Cell-Dyn Ruby and Humacount 5l / Estudio comparativo de los parámetros del eritrograma de Cell-Dyn Ruby y Humacount 5l

SUMMARY: The use of hematological counts for the prevention, diagnosis and follow-up of hematological diseases has increased. Indeed, the correct operation of a clinical laboratory is essential to producing comparable results. However, there is a paucity of validation and reproducibility studies among the different existing methods for clinical analysis. Therefore, our aim was to assess the commutability of the results provided by analyzers with different measuring systems. Sixty venous blood samples were obtained from patients, without discriminating for age or sex. Then, an automated hematological analysis was performed using the Cell-Dyn Ruby and HumaCount 5L instruments. The variables measured were: RBC, Hb, HCT, MCV, MCH and MCHC. The data were compared by a one-way ANOVA and Pearson's correlation coefficient. Statistical significance was fixed at p < 0.05. There were no statistically significant differences for RBC, HCT, MCH or MCHC. In addition, with the exception of MCHC, all the analytes showed a good correlation coefficient between the two instruments. There is a variety of automated systems for the clinical laboratory and it is essential for the clinician to know the different methodologies used in hematological analyzers as well as their sensitivity and specificity. Therefore, our results are useful for demonstrating the importance of practical knowledge of the analyzers mentioned.

RESUMEN: El uso de recuentos de células sanguíneas para la prevención, diagnóstico y monitoreo de enfermedades hematológicas ha ido en aumento. Por ello, el funcionamiento correcto de un laboratorio clínico es indispensable para producir resultados comparables. Sin embargo, existen pocos estudios de validación y reproducibilidad de los diferentes métodos de análisis clínico existentes. Por lo tanto, nuestro objetivo fue evaluar la intercambiabilidad de los resultados entregados por los analizadores que utilizan diferentes sistemas de medición. Se obtuvieron sesenta muestras de sangre venosa de pacientes, sin discriminar por edad o sexo. Los eritrogramas fueron obtenidos utilizando los analizadores automatizados Cell-Dyn Ruby y HumaCount 5L. Las variables medidas fueron: RBC, Hb, HCT, MCV, MCH y MCHC. Los datos fueron comparados por ANOVA a una vía y la correlación de Pearson. La significación estadística se estableció en el nivel estándar p<0,05. No hubo diferencias estadísticamente sig- nificativas para RBC, HCT, MCH y MCHC. Con la excepción de la MCHC, todos los analitos presentaron un buen coeficiente de correlación entre los dos analizadores comparados. Existen varios sistemas de automatización para su uso en laboratorios clínicos. Por lo tanto, es primordial para el clínico estar familiarizado con las diferentes metodologías utilizadas en los analizadores de sangre, así como su sensibilidad y especificidad. Nuestros resultados son útiles para mostrar la importancia del conocimiento práctico de los diferentes sistemas de medidas comparados.

Neutrophil scattering data driven pre-microscopic flagging of acute leukemic cases

ABSTRACT Background: The hematology analyzer, Sysmex XN-1000, generates white blood cell count with varying scattering intensities during a complete blood count (CBC) analysis. Objectives: The objectives of the study were to study the predictive role of median and coefficient of variation of neutrophil scattering items in blood samples for differentiation of leukemic subjects. Methods: We evaluated six neutrophil scattering parameters: neutrophil side scatter mean intensity, neutrophil side fluorescence light (SFL) mean intensity, neutrophil forward scatter mean intensity, neutrophil side scatter area distribution width (NE-WX), neutrophil SFL area distribution width (NE-WY), and neutrophil forward scatter area distribution width (NE-WZ), measured in white blood cell differential scattergram generated by the hematology analyzer (Sysmex XN-1000) at an academic medical center. Results: We collected 433 blood samples from acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) cases and normal controls. AML group showed highly significant differences in the mean values compared with the control group. Out of six neutrophil scattering items, NE-WX, NE-WY, and NE-WZ showed high efficiency, with area under the curve (AUC) values of 0.764, 0.748, and 0.757, respectively, to differentiate AML from ALL cases and control groups. When comparing combined acute leukemia cases (AML plus ALL) with the control group, NE-WX, NE-WY, and NE-WZ generated highly significant AUC values (0.840, 0.884, and 0.801, respectively). Conclusion: The neutrophil scattering parameters generated during CBC analysis provide a new tool for the prediction of acute leukemia and its lineage.

Inmunodeficiencias primarias y citopenias / Primary immunodeficiencies and cytopenias

Introducción: La visión actual de las enfermedades por inmunodeficiencia primaria (IDP) incluye un número creciente de síndromes que están asociados con la desregulación inmune y la autoinmunidad como características predominantes. Las citopenias autoinmunes pueden ser el primer signo de desregulación que precede a la presentación clásica de inmunodeficiencia primaria, con infecciones recurrentes u oportunistas. El conocimiento de un espectro de enfermedades potencialmente involucradas (hematológicas, reumatológicas e inmunológicas) es crucial para la identificación de una cierta proporción de genotipos y fenotipos de otros diagnósticos descritos. También permitirá excluir desórdenes como lupus eritematoso sistémico, inmunodeficiencia variable común, síndrome linfoproliferativo autoinmune; así como realizar diagnósticos diferenciales noveles como la deficiencia de GATA2, deficiencia de CD27, deficiencia de sensibilidad a lipopolisacáridos, síndrome fosfoinositol-3-quinasa delta activada, inmunodeficiencia ligada a X con déficit de magnesio y otros. Objetivo: Proporcionar una sinopsis conceptual de la aparición de citopenias en las IDP con el propósito de actualizar el conocimiento actual sobre dicho tema y de aumentar la percepción, tanto de hematólogos como inmunólogos, en relación a la presentación de citopenias como manifestación de estas enfermedades. Métodos: Se revisaron artículos originales y de corte experimental publicados en la década 2009 - 2019, en algunas bases de datos de la Biblioteca Virtual de Salud (BVS) de Cuba. Conclusiones: Al igual que las formas benignas autolimitadas de citopenia autoimmune post o parainfecciosas, o la neutropenia autoimmune adquirida de la infancia, que generalmente ocurren independientemente de una IDP subyacente reconocida, muchas de las citopenias que acompañan a esta enfermedad (pero no todas) están mediadas por autoanticuerpos. Es esencial entonces, que los médicos valoren, ante la evidencia clara de citopenia, que esta puede ser autoinmune(AU)

Introduction: The current view of primary immunodeficiency diseases (IDP) includes an increasing number of syndromes that are associated with immune dysregulation and autoimmunity as predominant characteristics. Autoimmune cytopenias may be the first sign of dysregulation that precedes the classic presentation of primary immunodeficiency, with recurrent or opportunistic infections. The knowledge of a spectrum of potentially involved diseases (hematological, rheumatological and immunological) is crucial for the identification of a certain proportion of genotypes and phenotypes of other diagnoses described. It will also allow excluding disorders such as systemic lupus erythematosus, common variable immunodeficiency, autoimmune lymphoproliferative syndrome; as well as making novel differential diagnoses such as GATA2 deficiency, CD27 deficiency, lipopolysaccharide sensitivity deficiency, activated delta phosphoinositol-3-kinase syndrome, X-linked immunodeficiency with magnesium deficiency and others. Objective: This review provides a conceptual synopsis of the appearance of cytopenias in the IDPs with the purpose of updating current knowledge on this topic and increasing the perception, of both hematologists and immunologists, in relation to the presentation of cytopenias as manifestation of these diseases. Methodos: Original and experimental articles published in the 2009-2019 decade were reviewed in some databases of the Virtual Health Library (VHL) of Cuba. Conclusions: As the self-limited benign forms of post or parainfectious autoimmune cytopenia, or childhood acquired autoimmune neutropenia, which generally occur independently of a recognized underlying IDP, many of the cytopenias that accompany this disease (but not all) mediated by autoantibodies. It is essential, then, that doctors assess, given the clear evidence of cytopenia, that it may be autoimmune(AU)

Blood cells profile in umbilical cord of late preterm and term newborns / Perfil de hemograma em sangue de cordão umbilical de recém-nascidos pré-termo tardio e a termo

ABSTRACT Objective: To describe the hematological profile in cord blood of late preterm and term newborns and compare blood indices according to sex, weight for gestational age and type of delivery. Methods: Cross-sectional study with late preterm and term newborns in a second-level maternity. Multiple gestation, chorioamnionitis, maternal or fetal hemorrhage, suspected congenital infection, 5-minute Apgar <6, congenital malformations, and Rh hemolytic disease were excluded. Percentiles 3, 5,10, 25, 50, 75, 90, 95 and 97 of blood indices were calculated for both groups. Results: 2,662 newborns were included in the sample, 51.1% males, 7.3% late preterms, 7.8% small for gestational age (SGA) and 81.2% adequate for gestational age (AGA). Mean gestational age was 35.6±1.9 and 39.3±1.0 weeks, respectively, for premature and term neonates. The erythrocytes indices and white blood cells increased from 34-36.9 to 37-41.9 weeks. Basophils and platelets remained constant during gestation. Premature neonates presented lower values ​​of all blood cells, except for lymphocytes and eosinophils. SGA neonates presented higher values ​​of hemoglobin, hematocrit and lower values of leukocytes, neutrophils, bands, segmented, eosinophils, monocytes and platelets. Male neonates presented similar values ​​of erythrocytes and hemoglobin and lower leukocytes, neutrophils, segmented and platelets. Neonates delivered by C-section had lower values ​​of red blood cells and platelets. Chronic or gestational hypertension induced lower number of platelets. Conclusions: Blood cells increased during gestation, except for platelets and basophils. SGA neonates had higher hemoglobin and hematocrit values and lower leukocytes. Number of platelets was smaller in male SGAs, born by C-section and whose mothers had hypertension.

RESUMO Objetivo: Descrever o perfil hematológico em sangue de cordão de recém-nascidos pré-termo tardio e a termo e comparar parâmetros hematimétricos segundo sexo, adequação peso idade gestacional e tipo de parto. Métodos: Estudo transversal com recém-nascidos pré-termo tardio e a termo, em maternidade de nível secundário. Excluíram-se gestação múltipla, corioamnionite, hemorragia materna ou fetal, suspeita de infecção congênita, Apgar no 5o minuto <6, malformações congênitas e doença hemolítica Rh. Calcularam-se os percentis 3, 5, 10, 25, 50, 75, 90, 95 e 97 dos parâmetros hematológicos. Resultados: Incluíram-se 2.662 recém-nascidos, 51,1% do sexo masculino, 7,3% prematuros tardios, 7,8% pequenos para a idade gestacional e 81,2% adequados. A idade gestacional foi 35,6±1,9 e 39,3±1,0 semanas, respectivamente, nos prematuros e termos. As séries vermelha e branca aumentaram de 34-36,9 para 37-41,9 semanas, exceto basófilos e plaquetas, que permaneceram constantes. Os prematuros apresentaram menores médias nas séries vermelha, plaquetária e branca, com exceção de linfócitos e eosinófilos. Recém-nascidos pequenos para a idade gestacional apresentaram maiores valores de hemoglobina e hematócrito e menores de leucócitos, neutrófilos, bastonetes segmentados, eosinófilos, monócitos e plaquetas. Recém-nascidos masculinos apresentaram taxas semelhantes de hemoglobina e hematócrito e menores de leucócitos, neutrófilos, segmentados e plaquetas. Na cesárea, as células vermelhas e as plaquetas foram menores que no parto vaginal. O número de plaquetas foi menor na hipertensão crônica ou gestacional. Conclusões: As células sanguíneas aumentaram durante a gestação, exceto plaquetas e basófilos. Recém-nascidos pequenos para a idade gestacional apresentaram maiores taxas de hemoglobina e hematócrito e menores de células brancas. O número de plaquetas foi menor no recém-nascido pequeno para a idade gestacional, masculino, nascido por cesárea e de mãe hipertensa.

Hemopoese em desnutrição proteica: caracterização do estroma medular e avaliação da participação do cálcio / Hematopoiesis in protein malnutrition: characterization of bone marrow stroma and evaluation of calcium participation

A desnutrição proteica continua sendo um dos principais problemas nutricionais do mundo. Trabalhos de nosso laboratório e de outros autores evidenciam que entre as alterações presentes na desnutrição proteica, está a alteração do tecido hemopoético, com modificações em componentes da matriz extracelular, alterações no ciclo celular da célula tronco/progenitora hemopoética, redução da produção de precursores hemopoéticos, tanto na série eritrocitária como na série leucocitária, levando a anemia e leucopenia. Os mecanismos de participação do Ca2+ nas células da medula óssea são pouco conhecidos, porém, sabe-se que ele atua no processo de hemopoese. Têm sido descrito que elevações da concentração de Ca2+ citoplasmático induzem a proliferação e diferenciação de células mielóides. A ação dessa via em indivíduos desnutridos também é pouco conhecida. Este estudo tem como objetivo avaliar o estabelecimento da celularidade medular in vitro, bem como investigar mecanismos moleculares envolvidos na proliferação e diferenciação dessa celularidade, além de avaliar a ação do cálcio na presença da interleucina-3 em células-tronco hemopoéticas murinas e sua modulação para avaliar alterações na via das MAPKs. Camundongos C57BL/6, machos e adultos foram submetidos à desnutrição proteica e, após a perda de aproximadamente 20% de seu peso corporal, as células da medula óssea foram colhidas. Essas células foram imunofenotipadas, além de reagirem com anticorpos específicos para caracterização da célula-tronco hemopoética e proteínas da via de sinalização de cálcio intracelular. Observamos que a celularidade do estroma medular em cultura de longa duração de animais desnutridos é alterada, principalmente em células de origem mesenquimal, que aparecem em maior número em desnutridos ao longo dos dias de cultura. Além disso, as ondas de cálcio intracelular estavam diminuídas em animais desnutridos, bem como as proteínas p-PKC, p-PLCy, CAMKII, p-AKT e p-STAT5 não respondem ao estímulo de IL-3, levando a uma deficiência da expressão das MAPK: ERK 1/2, JNK e p38. A desnutrição proteica pode causar alterações na celularidade estromal da medula óssea e na diferenciação das células tronco hemopoéticas pela via das MAPKs estimulada por IL-3

Protein malnutrition remains one of the world's major nutritional problems. Studies from our laboratory and others shown that alterations in protein malnutrition include hemopoietic tissue alterations, changes in extracellular matrix components, changes in the hemopoietic stem/progenitor cell tissue, reduction in the production of hemopoietic precursors, in the erythroid series as in the mieloyd series, leading to anemia and leukopenia. Mechanisms of Ca2+ participation in bone marrow cells are poorly understood, but no hemopoiesis has been developed. Elevations of cytoplasmic Ca2+ concentration in proliferation and differentiation of myeloid cells were included. Such an action through malnourished animals is also a little known. This study aims to evaluate the establishment of cellularity in vitro as well as investigate the molecular involvement in cell proliferation and differentiation, as well as to evaluate the action of calcium in the presence of IL-3 in hemopoietic stem cells and its modulation by analytical evaluations in the MAPKs pathway. C57BL/6, male adult mices were subjected to protein restriction and, after loss of approximately 20% of their body weight, bone marrow cells were harvested. These were immunophenotyped in addition to specific activation terms for the hemopoietic stem cell and intracellular signaling pathway proteins. We observed that the bone marrow cells in long-term culture of malnourished animals is altered, mainly in cells of mesenchymal origin, which appears in greater numbers in undernourished throughout the days of culture. In addition, as intracellular calcium waves decreased in malnourished animals, as well as the p-PKC, p-PLC, CAMKII, p-AKT and p-STAT5 proteins did not respond to IL-3, sugesting expression of the expression of MAPK: ERK 1/2, JNK and p38. Protein malnutrition may have changes in bone marrow capacity and differentiation of hemopoietic stem cells through IL-3-stimulated MAPKs

Establishment of Age- and Gender-Specific Reference Ranges for 36 Routine and 57 Cell Population Data Items in a New Automated Blood Cell Analyzer, Sysmex XN-2000

We established age- and gender-specific reference ranges for the 36 routine complete blood cell (CBC) and 57 cell population data (CPD) items in the Sysmex XN-2000 (Sysmex, Japan). In total, 280 peripheral blood samples were obtained from an equal number of healthy adults. Values for 36 routine items and 57 CPD items were obtained for each sample, and the results were categorized into six subgroups (N>39 in each subgroup) according to patient age (20-40, 41-60, and >60 yr) and gender (male and female), and compared with respect to age and gender differences. The majority of data items (22 of 36 routine CBC items and 44 of 57 CPD items) exhibited significant differences (P< or =0.05) in their results with respect to age or gender, and several red cell-, lymphocyte-, and platelet-related data tended to decrease in women or older adults. These results provide a basis for establishing age- and gender-specific reference ranges for routine and CPD items in Sysmex XN-2000. Furthermore, these reference ranges could be used to determine clinical significance for new items of Sysmex XN-2000 in further studies.

La Biometría hemática desde la optica de la homeopatía: hemograma, citometríahemática, citología hemática / The biometrics from the viewpoint of homeopathy: CBC, citometría hemática, hematic cytology

Las pruebas de laboratorio son un complemento importante para que el médico homeópata recopile la totalidad de los datos con que se manifiesta la enfermedad de sus pacientes. Uno de los exámenes más habituales a los que se puede recurrir es la biometría hemática, el cual ofrece una orientación conveniente sobre los mecanismos que se activan en el organismo ante un estímulo nocivo determinado,mediante información detallada sobre los elementos formes de la sangre: serie blanca, serie roja y plaquetas. Se presentan a continuación las consideraciones más importantes sobre las variables que se estudian mediante un hemograma, pues aunque ninguno delos valores que nos puede proporcionar se encuentra en los repertorios clásicos,es evidente que nos ofrece información sobre signos tan importantes o comunescomo anemia, hemorragias, trombosis, tromboflebitis o abscesos, todos ellos ensus diferentes modalidades.

Laboratory tests are an important complement to the homeopath. They help to collect all the data manifested by the patient´s disease. One of the most common tests to which the homeopath can turn to is the Cell Blood Count, which offers a convenient guidance of the mechanisms that are activated in the body to a determined noxious stimulus, through detailed information about the blood cells: white,red and platelets count.The following are the most important considerations of the variables that are studied by a CBC, as though none of the values that we can find is in the classical repertoire, it is evident that offers very important information on common signs such as anemia, bleeding, thrombosis, thrombophlebitis or abscess, all in their different ways.

Avances y aplicación clínica de la citometría hemática automatizada / Advances and clinical application of automated hematic cytometry

Se revisan los antecedentes históricos del surgimiento de la automatización en la rama hematológica y los principios de detección utilizados por los contadores automáticos de células sanguíneas, con referencia a su acción combinada en la caracterización de las distintas poblaciones celulares hemáticas. Se describen los componentes básicos de los analizadores hematológicos. Se detalla la obtención, intervalo de referencia y utilidad clínica de las variables de uso común ofrecidas por estos equipos en el estudio de los eritrocitos y trombocitos, referidos a los parámetros más actualizados en el estudio de estas series celulares. En cuanto a los leucocitos, se mencionan los principales parámetros relacionados con el estudio automatizado de uso rutinario, la utilidad de los novedosos principios inmunológicos de detección en la caracterización de esta población celular, así como los logros y obstáculos manifiestos durante el conteo y estudio de las subpoblaciones leucocitarias inmaduras o anómalas. Por último, se exponen las ventajas del uso de los contadores hematológicos para el trabajo actual del laboratorio de hematología y la necesidad del conocimiento de sus potencialidades diagnósticas por parte de los médicos, especialmente los hematólogos e inmunólogos

Historical background on the development of automation in hematology as well as the major detection principles used by automated blood cell counters are reviewed, referring to their combined action in the characterization of various hematic cell populations. The basic components of hematological analyzers are described. Obtention reference interval and clinical utility of commonly used variables provided by these equipments for the study of erythrocytes and thrombocytes are described referring to the most updated parameters for the study of these cell series. The main parameters of the everyday leukocyte automated study, the novel immunological detection principles used in its characterization, as well as the achievements and obstacles found during this process are mentioned. Finally, the advantages of blood cell counters for the current hematology laboratory work and the need of knowledge of its diagnostic potential by physicians, particularly hematologists and immunologists, are presented

Validation of the Sysmex sp-1000i automated slide preparer-stainer in a clinical laboratory

Background: The speed and quality of information have become essential items in the release of laboratory reports. The Sysmex®SP1000-I device has been developed to prepare and stain smear slides. However, for a device to be cleared for use in the laboratory routine it must pass through a validation process. Objective: To evaluate the performance and reliability of the Sysmex® SP-1000i slide preparer-stainer incorporated into the routine of a hospital laboratory in Porto Alegre. Methods: Peripheral blood samples of patients attending the laboratory for ambulatory exams with leukocyte counts between 7000/°L and 12,000/°L were evaluated, independent of gender and age. Two slides were prepared for each sample using the Sysmex® SP-1000i equipment; one of the slides was used to perform quality control tests using the CellaVision® DM96 device, and the other slide was used to compare pre-classification by the same device and the classification performed by a pharmacist-biochemist. Results: The results of all the slides used as controls were acceptable according to the quality control test as established by the manufacturer of the device. In the comparison between the automated pre-classification and the classification made by the professional, there was an acceptable variation in the differential counts of leukocytes for 90% of the analyzed slides. Pearson correlation coefficient showed a strong correlation for band neutrophils (r = 0.802; p-value < 0.001), segmented neutrophils (r = 0.963; p-value < 0.001), eosinophils (r = 0.958; p-value < 0.001), lymphocytes (r = 0.985; p-value < 0.001) and atypical lymphocytes (r = 0.866; p-value < 0.001) using both methods. The red blood cell analysis was adequate for all slides analyzed by the equipment and by the professional. Conclusion: The new Sysmex®SP1000-i methodology was found to be reliable, fast and safe for the routines of medium ...

Avaliando a comutatividade: importante requisito da qualidade para laboratórios clínicos / Understanding commutability: important quality requirement for clinical laboratories

INTRODUÇÃO: Poucos laboratórios no Brasil realizam a avaliação da comutatividade dos resultados de exames, provavelmente por desconhecimento dos procedimentos para sua realização e também pela pouca importância dada a essa avaliação. OBJETIVO: Avaliar a comutatividade dos resultados de exames hematológicos realizados em três analisadores automatizados em um laboratório de um hospital público universitário, em Belo Horizonte, MG, Brasil, propondo procedimento prático, de baixa complexidade, factível de ser utilizado em laboratórios clínicos. MATERIAL E MÉTODOS: As amostras foram selecionadas visando obter valores nos níveis de decisão terapêutica. Foram processadas seis amostras por dia, durante quatro dias, em duplicata em três analisadores, totalizando 48 replicatas em cada instrumento. Foi avaliada a correlação entre os resultados de 10 dos principais parâmetros hematológicos obtidos nos instrumentos-teste e aqueles obtidos com o instrumento-referência. Foram estimados os erros sistemáticos e totais, considerando as especificações da variação biológica como limite máximo aceitável. RESULTADOS: O coeficiente de correlação (r) entre os equipamentos para os parâmetros investigados foi > 0,975. Os erros sistemático (médio) e total, obtidos para os parâmetros analisados, quando se comparam os equipamentos de teste com o de referência, atenderam às especificações da qualidade definidos. Discussão: A comutatividade é um importante processo da gestão da qualidade do laboratório clínico e visa garantir a comparabilidade de resultados de exames realizados por diferentes sistemas. CONCLUSÃO: Utilizando procedimento padronizado internacionalmente, prático e de baixa complexidade, demonstrou-se que os exames realizados nos equipamentos avaliados são equivalentes, podendo ser usados indistintamente no acompanhamento de pacientes.

INTRODUCTION: Few Brazilian laboratories evaluate the commutability of test results, probably due to the lack of procedure expertise as well as the neglected importance of its assessment. OBJECTIVE: The aim of this study was to evaluate the commutability of hematological test results performed with three automated analyzers at the laboratory of a public university hospital in Belo Horizonte, MG, Brazil, proposing a practical, simple, and feasible procedure to be applied in clinical laboratories. MATERIAL AND METHODS: Samples were selected in order to obtain hematologic values for therapeutic decision levels. Six samples were processed in duplicate in three analyzers daily during a four-day period amounting to a total of 48 replicates in each instrument. The correlation between the results of 10 hematologic parameters obtained with test instruments and the reference instrument was assessed. Systematic and total errors were estimated and criteria for acceptable performance were based on the biological variation specifications. RESULTS: The correlation coefficient (r) between test instruments and reference instrument results was > 0.975. Systematic (mean) and total errors met the required quality specifications when compared with reference instruments. Discussion: Commutability is an important process of quality management in clinical laboratories and it ensures the comparability of test results carried out with different procedures. CONCLUSION: Through a practical, simple, and internationally standardized procedure, this study showed that test results from the evaluated instruments were equivalent, which allows their use in patient monitoring.

Ultrastructural characterization of bovine umbilical cord blood cells / Caracterização ultra-estrutural das células sanguíneas do cordão umbilical bovino

The umbilical cord blood (UCB) is an important source of pluripotent stem cells, which motivated researches on ontogeny and transplantation. The morphological characterization of umbilical cord cells is the first step to establish subsequent experiments on these areas. Although some information on humans can be found, no data on UCB is available for bovines. Therefore, this work is the first attempt to conduct an ultrastructural characterization of bovine umbilical cord blood. Blood was collected from the umbilical cord of twenty fetuses by punction of the umbilical vein. Samples were processed for whole leucocytes observation by centrifugation and the buffy coat was collected. Cells were washed and pelleted and prepared according to the standard protocol of the transmission electron microscopy. The presence of cells with morphologic characteristics compatible with the precursors from the erythrocytic, neutrophilic, eosinophilic, basophilic, and lymphocytic lineages was observed. Atypical cells with peculiar morphological features, strongly similar to apoptotic cells, were seen. Bovine neutrophils with three types of cytoplasmic granules were also found in the blood. The ultrastructural characteristics of observed bovine UCB cells where similar to those found in other species, suggesting that bovines could possibly constitute an experimental model for approaches on UCB cells research.

O sangue de cordão umbilical (SCU) é uma importante fonte de células progenitoras pluripotentes, que motiva pesquisas em ontogenia e transplantes. A caracterização morfológica das células de cordão umbilical é o primeiro passo para se estabelecer experimentos subsequentes nessas áreas. Embora algumas informações sobre SCU em humanos possam ser encontradas, não existe nenhuma informação disponível sobre elas em bovinos. Portanto, este trabalho é a primeira tentativa de se conduzir uma caracterização ultra-estrutural do sangue de cordão umbilical bovino. O sangue foi coletado do cordão umbilical de 20 fetos por punção da veia umbilical. As amostras foram processadas para observação dos leucócitos totais por centrifugação pela coleta do botão leucocitário. As células foram lavadas, peletizadas e preparadas de acordo com protocolo padrão para microscopia eletrônica de transmissão. A presença de células com características morfológicas compatíveis com precursores das linhagens eritrocítica, neutrofílica, eosinofílica, basofílica e linfocítica foram observadas. Células atípicas com características morfológicas peculiares muito semelhantes a células em apoptose foram observadas. No sangue do cordão umbilical também foi encontrado neutrófilos bovinos apresentando três tipos de grânulos citoplasmáticos. As características ultraestruturais do SCU bovino foram semelhantes às encontradas em outras espécies, sugerindo que esta espécie possa servir como modelo experimental para abordagens em pesquisa sobre sangue de cordão umbilical.

Necesidad de la evaluación morfológica del hemograma en un hospital complejo tipo 1: control de calidad y gestión / Necessity of the morphological evaluation of the haemogram in a complex hospital type 1: management and quality control

Objetivo: Determinar criterios de selección de muestras para la evaluación morfológica del hemograma, basados en parámetros clínicos y resultados del contador hematológico y en términos de gestión, costos y clínica, los resultados de un programa de selección de muestras para la evaluación morfológica del examen en un hospital tipo 1. Pacientes, Materiales y Método: Pacientes con una muestra tomada en el Hospital de Carabineros de Chile durante el mes de Agosto de 2009, las que fueron analizadas en un equipo Beckman Coulter HmX@ y evaluadas morfológicamente por el Tecnólogo Médico. Se registraron los parámetros clínicos tales como hemoglobina, leucocitos y plaquetas, además del lugar de derivación, especialidad y edad del paciente. Los resultados fueron analizados, tanto por alerta clínica como instrumental, incluyendo todos los parámetros considerados clínicamente importantes. Resultados: Participaron 2.222 pacientes, hubo concordancia entre el Tecnólogo Médico y el analizador en 93,9 por ciento de los casos, con una sensibilidad de 97,3 por ciento y una especificidad de 88,9 por ciento con valores predictivos positivo y negativo de 95,9 por ciento y 92,7 por ciento respectivamente, para detectar una muestra anormal. Por criterios de selección se eliminó la evaluación morfológica en 79,5 por ciento de los casos. Hubo 1,6 por ciento de casos falso-negativos en los criterios de morfología. Discusión: El estudio demostró que es posible disminuir el número de evaluaciones morfológicas hechas al hemograma en un 80 por ciento de los casos, lo cual es semejante a los estudios internacionales efectuados en un hospital tipo 1. El sistema de alerta clínica pone énfasis en la importancia de que el médico tratante ingrese la información clínica en la solicitud de hemograma. Hay una liberación importante de tiempo de Tecnólogo Médico para utilizar su capacidad en funciones de mayor complejidad.

Objectives: To determine selection criteria for the morphological evaluation of full blood count samples, based on clinical criterias and the results of an automatic hematological analyzer. To analyze the cost-benefits and gestion of such a selection procedure in samples obtained from a public general hospital. Patients, Materials and Methods: AII patients with a full blood count taken at the Hospital de Carabineros de Chile during August 2009 were included AII samples were analyzed using a Beckman-Coulter HmX@ automatic analyzer and analyzed morphologically by the Medical Science Officer. Patient details including age, sex, hematological parameters Hemoglobin, white cell and platelet count and place of reference were noted the results were analyzed using clinical and instrument alerts, including all clinical hematological parameters considered important. Results: 2,222 patients were included in the study. There were an agreement in analysis between the HmX and Medical Science Officer in 93.9 percent of cases, with a sensitivity of 93.9 percent specificity of 88.9 percent, a positive and negative predictive value of 95.9 percent and 927 percent respectively to detect and abnormal blood count. Owing to the selection criteria used it was possible to eliminate the need for morphological evaluation in 79.5 percent of cases, the possibility of a serious error of omission being minimal, only 1,6 percent of cases were false negative, on morphological and not clinical grounds. Discussion: The study showed that it is possible to decrease the number of morphological evaluations in 80 percent of cases, which is similar to the figures internationally reported for a general hospital. The clinical alert puts emphasis on the treating physician to complete clinical details on the order form. In total, there were 1.6 percent of false negative cases on morphological grounds y it is important to note that some morphological abnormalities will not be detected the...

Variação da contagem de células somáticas em vacas leiteiras de acordo com patógenos da mastite / Somatic cell counts variation in dairy cows according to mastitis pathogens

Avaliou-se o efeito de patógenos da mastite sobre a contagem de células somáticas (CCS) em leite. Foram coletadas 3.987 amostras de leite de 2.657 animais oriundos de 24 rebanhos leiteiros localizados nos estados do Rio de Janeiro e Minas Gerais. As amostras de leite foram usadas para CCS e identificação de patógenos da mastite. Estatísticas descritivas, teste T para amostras independentes e modelo linear generalizado foram usados para análise dos dados. O modelo linear generalizado identificou os efeitos de rebanho, animal dentro de rebanho, ordem de parto, estação do ano e infecção intramamária causada por Streptococcus agalactiae e Streptococcus spp. que não S. agalactiae como significativos na variação da CCS. O efeito de animal dentro de rebanho foi maior que o efeito de rebanho. S. agalactiae foi o patógeno responsável pelo maior aumento da CCS em vacas e apresentou em média 1.520.000 células/mL. Foi observado efeito específico dos patógenos na variação da CCS.

The influence of mastitis pathogens on variation of milk somatic cell count (SCC) was evaluated. Three thousand nine hundred eighty-seven milk samples were colected from 2,657 dairy cows in 24 herds located in the states of Minas Gerais and Rio de Janeiro. The milk samples were used to SCC and identification of mastitis pathogens. Descriptive statistics, T test for independent samples, and generalized linear model were used to data analysis. The generalized linear model identified the effects of herd, animal within herd, parity, year season, intramammary infection, and infection caused by Streptococcus agalactiae and Streptococcus spp. except S. agalactiae as significant on SCC variation. The effect of animal within herd was higher than the effect of herd. S. agalactiae was the pathogen responsible for higher SCC increasing and presented the average of 1,520,000 cells/mL. The specific effect on SCC variation was observed in the study.

Hemograma, proteinograma, ionograma e dosagens bioquímicas e enzimáticas de ovinos acometidos por conidiobolomicose no nordeste do Brasil: [revisão] / Hematology and blood biochemistry in sheep with conidiobolomycosis in northeastern Brazil: [revision]

Foram realizados eritrogramas, leucogramas, dosagens séricas de proteínas totais, albumina, globulinas, cálcio, fósforo, magnésio, cloretos, uréia, creatinina, bilirrubina total, direta e indireta e atividade sérica de aspartato aminotransferase (AST), fosfatase alcalina (FA) e gama-glutamiltransferase (GGT) de 56 ovinos afetados por conidiobolomicose e de 371 ovinos sadios provenientes dos mesmos rebanhos dos casos da doença. Os resultados revelaram que os ovinos com conidiobolomicose apresentam anemia arregenerativa normocítica normocrômica, leucocitose com neutrofilia e discreto desvio à esquerda regenerativo, com elevação da relação neutrófilo:linfócito, monocitose moderada, trombocitose, hipoproteinemia (hipoglobulinemia), hipomagnesemia, hipocalcemia e elevação dos teores de fósforo. As atividades séricas de AST e GGT estavam aumentadas e as de FA diminuídas. Não houve alteração nos níveis de uréia e creatinina, mas ocorreu hipoglicemia e hiperbilirrubinemia. Estes resultados podem ser utilizados para estudos experimentais da doença, em ensaios com tratamentos e para identificar casos precoces da enfermidade que não apresentem sinais clínicos. Além disso, os valores encontrados em 371 ovinos deslanados hígidos podem ser utilizados como valores referenciais para este tipo de ovinos na região semi-árida do Brasil.

Hematologic values, serum levels of albumin, globulins, total proteins, calcium, phosphorus, magnesium, chloride, urea nitrogen, creatinine, total, direct and indirect bilirrubin, and serum activities of aspartate aminotransferase (AST), alkalin phosphatase (AF) and gama-glutamyltransferase (GGT) were determined in 56 hair sheep with conidiobolomycosis and 371 healthy hair sheep from the same flocks. Sheep with conidiobolomycosis had nonregenerative, normocytic and normochromic anemia, leucocytosis with neutrophilia and increased nutrophil:lymphocyte rate, moderate monocytosis, moderate thrombocytosis, hypoproteinemia (hypoglobulinemia), hypomagnesemia, hypocalcemia and increased values of phosphorus. Serum activities of AST and GGT were increased and serum activity of FA decreased. Serum values of urea and creatinine were within normal values. Hypoglycemia and hyperbilirrubinemia were also observed. These results can be used for experimental studies with the disease, for treatments trials, and to detect early cases of the disease without clinical signs. Otherwise the hematologic and blood biochemistry values of healthy sheep can be used as reference values for hair sheep in the semi-arid region of Brazil.

Microrganismos patogênicos, celularidade e resíduos de antimicrobianos no leite bovino produzido no sistema orgânico: [revisão] / Pathogenic microorganisms, somatic cell count and drug residues evaluation in organic bovine milk: [revision]

Nos últimos anos cresce a preocupação dos consumidores quanto à qualidade do leite e às condições de produção e bem-estar dos animais. Simultaneamente, aumenta o interesse e o consumo de produtos e subprodutosde origem animal produzidos no sistema orgânico, com destaque para o leite e derivados. O presente estudo investigoua presença de microrganismos patogênicos, a sensibilidade e a multi-resistência dos isolados aos antimicrobianos, a celularidade e a presença de resíduos de drogas no leite de vacas, com e sem mastite, produzido no sistema orgânico. Foram amostradas 148 vacas no período médio de lactação, das quais duas com mastite clínica, 72 com mastite subclínica e 74 sem mastite (controles), provenientes de quatro pequenas propriedades do interior do Estado de São Paulo, certificadas como orgânicas. Staphylococcus aureus (25,7%), Streptococcus spp. (21,4%), Corynebacterium bovis (12,9%), Streptococcus agalactiae (4,3%) e Staphylococcus spp. (4,3%) foram os microrganismos mais frequentemente isolados nos animais com mastite. Aspergillus spp. foi isolado de um animal. Ceftiofur (95,2%), oxacilina (84,2%), gentamicina (76,3%) e cefoperazona (70,3%) foram os antimicrobianos mais efetivos frenteaos isolados. As maiores taxas de resistência das linhagens foram constatadas para penicilina (53,5%), ampicilina (41,6%) e neomicina (38,6%). Multi-resistência a trêsou mais fármacos foi encontrada em 40 (39,6%) linhagens. A média da contagem de células somáticas das propriedades para animais com mastite foi 175.742,67 céls/mL, enquantoque para os animais controles foi 58.227,6 céls/mL. A presença de resíduos de antimicrobianos foi observada no leite de quatro (2,7%) animais. Os resultados revelarambaixa celularidade média nos animais, indicativo de boa qualidade do leite. Entretanto, apontam para a necessidadeda adoção de medidas de controle para microrganismos contagiosos e maior...

In last years increase the importance of milk quality and conditions of bovine milking. Simultaneously, increase the interest about organic milk and derivates. The aim of present study was investigate the milk pathogens, sensitivity and multiple drug resistance of isolates, somatic cell count and residues of drugs in milk, from cattle with and without mastitis, come from four little organic dairy farms in State of São Paulo, Brazil. Were used 148 cattle on the middle period of lactation. From these, two showed clinical mastitis, 72 subclinical mastitis and 74 without signs of mammary inflammation (controls). Staphylococcus aureus (25.7%), Streptococcus spp. (21.4%), Corynebacterium bovis (12.9%), Streptococcus agalactiae (4.3%) and Staphylococcus spp. (4.3%) were the more-frequent microorganismsisolated from animals with mastitis. Aspergillus spp. was isolated from one animal. Ceftiofur (95.2%), oxacillin (84.2%), gentamicin (76.3%) and cefoperazone (70.3%) were the moreeffective drugs. High resistance of isolates were found to penicillin (53.5%), ampicillin (41.6%) and neomycin (38.6%). Multiple drug resistance to three or more drugs was observed in 40 (39.6%) isolates. Media of somatic cell count encountered in animals with mastitis and controls were 175,742.67cs/mL and 58,227.6 cs/mL, respectively. Antimicrobials residuesin milk were detected in four (2.7%) animals. The present findings showed the low somatic cell count of animals, indicative of good quality of milk. However, pointed the need of control measures for contagious pathogens of bovine mastitis and more attention forprohibition of antimicrobial use in organic dairy farms.

Hemograma e bioquímica sérica de tartarugas cabeçudas (Caretta caretta) de vida livre e mantidas em cativeiro, no litoral norte da Bahia / Hemogram and serum biochemistry of free-ranging and captive loggerhead sea turtles (caretta caretta), from north coast of Bahia, Brazil

Com o objetivo de se obter dados que auxiliem na rotina clínica de tartarugas marinhas, realizou-se o hemograma e a determinação de alguns parâmetros bioquímicos séricos de fêmeas de tartarugas cabeçudas (Caretta caretta) de vida livre (n = 22), animais que desovam no Litoral Norte da Bahia, e em cativeiro (n = 5), animais mantidos no Centro de Visitantes do Projeto Tamar-ICMBio, na Praia do Forte, Brasil. Os valores obtidos para os dois grupos foram comparados, onde se observou diferença estatística significativa (p<0,05) para as contagens relativas e absolutas de eosinófilos e contagem relativa de monócitos dentre as variáveis do hemograma e, para a proteína total, globulina, glicose, colesterol e aspartato aminotransferase. Possivelmente, diferenças nas condições ambientais, nutricionais e reprodutivas, as quais estes animais foram submetidos, influenciaram nos resultados.

The aim of this study was to improve the clinical evaluation in sea turtles by analyzing the hemogram and serum biochemistry in female logger head sea turtles (Caretta caretta), free-ranging (n=22) nesting in the north coast of Bahia and in the animals captivity (n=5) in the visitor center of the Projeto Tamar-ICMBio, in Praia do Forte, Bahia, Brazil. The values obtained were compared between the two groups and it was observed that there was statistically significant difference(p<0,05) for the relative and absolute eosinophil count, relative monocyte count, total serum protein, globulin, glucose, cholesterol and aspartate aminotransferase. The results variation is probably related to the differences in environmental, nutritional and reproductive conditions