Immature granulocytes index as early marker of sepsis.

INTRODUCTION: Sepsis induces the recruitment of immature neutrophils into the circulation. An immature granulocyte percentage (IG%) count greater than 3% has been shown to be an indicator for the risk of sepsis. The aim of this study was to evaluate the IG% as predictor of sepsis compared to blood culture results and sepsis diagnostic confirmation. METHODS: The study included individuals (n = 301) of both sexes aged ≥18 years who underwent Hospital São Lucas examinations between January and November 2017. For all the patients, IG%, as well as blood culture results, were evaluated. All examinations were obtained from Clinical Laboratory database. Data were analyzed through the SPSS program version 18.0. RESULTS: There was statistical association between blood culture and IG% results (P = 0.009) and between sepsis confirmation and IG% on Pearson chi-square test (P < 0.001). An IG% cutoff point of 2.0% was able to exclude sepsis based on clinical diagnosis with a specificity of 90.9% and a sensitivity of 38.5%. The cutoff value in ROC analyses of IG% based on blood culture results was 0.3% and 0.4% based on clinical diagnosis. CONCLUSION: Our study demonstrated that IG% <2.0% are helpful on the exclusion of sepsis diagnosis with a very high specificity (90.9%). The IG% is a useful additional marker for sepsis diagnosis allowing the early initiation of therapy and better possibilities of recovery.

Mean Platelet Volume and Immature Platelet Fraction in Autoimmune Disorders.

Mean platelet volume (MPV), measured using automated blood analysers, has been appraised as a potential biomarker in cardiovascular disease, diabetes mellitus, and cancer. The test, a useful tool in differentiation of thrombocytopenic states, has now been carried out for autoimmune disorders, but data are yet scarce. Controversial results have been obtained in systemic and organ-specific autoimmune disorders. Another test, the immature platelet fraction (IPF) reflects the amount of young, reticulated platelets. IPF is calculated by automated hematology analysis or flow cytometry, and it is usually high in patients with rapid platelet destruction. For both MPV and IPF, standardization of cutoff is a major need. In this review, we focus the current applicability of MPV and IPF as biomarkers in patients with autoimmune diseases.

Immature platelet fraction in hypertensive pregnancy.

BACKGROUND: Imbalance in hemostatic mechanisms can occur during pregnancy with a tendency for hypercoagulability and increased thrombosis risk. Pregnant women with hypertensive disorder, especially preeclampsia, show alterations in platelet indexes. Immature platelet fraction (IPF) has been suggested as a sensitive index for monitoring changes in platelet production and destruction. OBJECTIVES: To evaluate the IPF in patients diagnosed with a gestational hypertensive disorder (GHD). PATIENTS AND METHODS: A cross-sectional study was conducted at an University Hospital to estimate maternal blood IPF index in 99 pregnant women, divided into three groups: normotensive pregnancy (NP), preeclampsia syndrome (PES), and non-proteinuric hypertensive pregnancy (nPHP). Following ethical approval and written informed consent, samples were collected from 33 NP, 34 PES, and 32 nPHP women. Platelet indexes were measured by fluorescent flow cytometry. RESULTS: IPF and mean platelet volume (MPV) counts in GHD were significantly higher than in NP (IPF: 3.8, 2.4-5.1%; 8.6, 5.8-10.6%; 7.3, 4.2-10.2%; p < 0.001 and MPV: 10.6 ± 0.9 fL; 12.1 ± 1.0 fL; 11.6 ± 1.0 fL; p < 0.001 for NP, PES, and nPHP, respectively). No difference was detected between PES and nPHP groups. The distribution of patients with an IPF above 6.1%for NP, PES, and nPHP was 9%, 65%, and 43.8%, respectively (p < 0.001). IPF as a test to differentiate GHD from the controls achieved an area under the curve of 0.83 on a receiver operating characteristics curve. CONCLUSION: A distinct profile in platelet indexes was detected in hypertensive pregnancies. It suggests that these markers could be used in daily routine as an additional tool in the management of pregnant women.

New red blood cell and reticulocyte parameters and reference values for healthy individuals and in chronic kidney disease / Novos parâmetros e valores de referência eritrocitários e reticulocitários normais e na doença renal crônica

Introduction: The importance of local references values has been well described in the literature; this is because the characteristics of the population may influence the laboratory tests. Objective: To establish the reference range for traditional and extended red blood cell parameters and reticulocyte indices in order to investigate its application in patients with chronic kidney disease (CKD). Materials and methods: 249 blood donors (125 males and 124 females) were selected to establish the reference values. The hemodialysis sample consisted of 62 patients with terminal CKD (48 male and 14 female). All analyzes were performed using the Sysmex XE-5000 automated hematology analyzer. Results: Differences between reference values was observed in relation to gender: red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular hemoglobin concentration (MCHC), percentage of hyperchromic red blood cells (%HYPER), percentage of microcytosis (%MICRO), percentage of macrocytosis (%MACRO), absolute reticulocyte count (RET), reticulocyte hemoglobin content (RET-He), immature reticulocyte fraction (IFR), low fluorescence reticulocytes (LFR), medium fluorescence reticulocytes (MFR), and high fluorescence reticulocytes (HFR). Individuals with CKD presented RBC, HGB, HCT, MCHC, red cell distribution width expressed as coefficient of variation (RDW-CV), percentage of hypochromic red blood cells (%HYPO), percentage of reticulocytes (RET%), RET (female group), IFR, LFR, MFR, and HFR results compatible with the anemic state, which can be observed in 91.8% of patients. All studied parameters were in the area under the curve (AUC) > 0.4. In male group, %HYPO (AUC: 0.806) and IFR (AUC: 0.762) presented higher AUC values, while female group presented %HYPO (AUC: 0.806), %HYPER (AUC: 0.815), and IFR (AUC: 0.660). Conclusion: The medical advancement, the development of new techniques and hematological parameters have revealed important information ...

Introdução: A importância dos valores de referências locais tem sido bastante descrita na literatura, isso porque características da população podem influenciar os testes laboratoriais. Objetivos: Estabelecer o intervalo de referência para parâmetros eritrocitários tradicionais e estendidos e índices reticulocitários a fim de investigar sua aplicação em pacientes com doença renal crônica (DRC). Materiais e métodos: Dos doadores de sangue, 249 pacientes foram selecionados para estabelecimento dos valores de referência (125 homens e 124 mulheres); dos pacientes em hemodiálise, a amostra foi composta por 62 indivíduos com DRC terminal (48 homens e 14 mulheres). Todas as análises foram realizadas no avaliador hematológico Sysmex XE-5000. Resultados: Foi observada uma distinção entre os valores de referência em relação ao gênero: células vermelhas do sangue (RBC), hemoglobina (HGB), hematócrito (HCT), concentração de hemoglobina corpuscular média (CHCM), porcentagem de eritrócitos hiper-hemoglobinizados (%HIPER), porcentagem de microcitose (%MICRO), porcentagem de macrocitose (%MACRO), contagem absoluta de reticulócitos (RET), contagem relativa de reticulócitos (RET-He), fração de reticulócitos imaturos (IFR), reticulócitos de baixa fluorescência (LFR), reticulócitos de média fluorescência (MFR) e reticulócitos de alta fluorescência (HFR). Os indivíduos com DRC apresentaram resultados de RBC, HGB, HCT, CHCM, coeficiente de variação do tamanho dos eritrócitos (RDW-CV), %HIPO, RET%, RET (grupo das mulheres), IFR, LFR, MFR e HFR compatíveis com o estado anêmico, que pode ser observado em 91,8%. Todos os parâmetros estudados apresentaram área sob a curva (AUC) > 0,4. Para o grupo dos homens, a %HIPO (AUC: 0,806) e a IFR (AUC: 0,762) apresentaram melhores valores de AUC; já para o grupo das mulheres foram a %HIPO (AUC: 0,806), a %HIPER (AUC: 0,815) e a IFR (AUC: 0,660). Conclusão: Avanço da medicina e surgimento de novas técnicas e parâmetros ...

Marcador de risco hemostático associado a eventos cardiovasculares em síndrome metabólica / Hemostatic risk marker associated with cardiovascular events in metabolic syndrome

Fundamentos: A doença cardiovascular (DCV) é a principal causa de morte nos países em desenvolvimento. Os indivíduos com síndrome metabólica (SM) apresentam risco elevado para DCV. Os fatores de risco tradicionais em conjunto, não explicam todos os eventos cardiovasculares. O fator von Willebrand (FvW), envolvido na agregação plaquetária e trombose, tem sido investigado nesse contexto. Objetivo: Investigar a relação entre FvW e DCV, em pacientes portadores de SM, com e sem eventos cardiovasculares prévios. Métodos: Estudados 77 pacientes ambulatoriais, ≥18 anos, portadores de SM de acordo com os critérios estabelecidos pela NCEP-ATP III. Mediu-se o nível plasmático do FvW e comparou-se os valores médios entre os grupos com DCV prévia (n=30) e sem DCV documentada (n=47). Resultados: Da população estudada, 66,0% eram do sexo feminino, 78,0% de etnia branca, média de idade 63,7±8,9 anos, peso médio 82,9±14,9 kg e índice de massa corpórea 32,2±4,8 kg/m2. O nível plasmático médio do FvW foi similar nos pacientes com e sem DCV prévia, com valores de 154,5±52,1 e 155,47±41,4, respectivamente. Observou-se associação entre o diabetes mellitus (DM) e DCV estabelecida, que permaneceu significativa após ajuste para as demais variáveis incluídas no modelo multivariado. Conclusões: Não houve diferença no nível plasmático médio do FvW entre os pacientes portadores de SM, com e sem DCV documentada. A presença de DM, entretanto, esteve associada de forma independente com DCV nesta população.

Background: Cardiovascular disease (CVD) is the leading cause of death in developing countries. Individuals with metabolic syndrome (MS) are at increased risk for CVD. The traditional risk factors, altogether, do not explain all cardiovascular events. The von Willebrand factor (vWF), involved in platelet aggregation and thrombosis, has been investigated in this context. Objective: To investigate the relationship between the vWF and CVD in patients with MS, with and without previous cardiovascular events. Methods: The study included 77 outpatients, ≥18 years, with MS, according to the criteria established by NCEP-ATP III. The plasma level of vWF was measured and the mean values were compared between the groups with prior CVD (n=30) and without documented CVD (n=47). Results: In the study population, 66.0% were female, 78.0% were white, mean age 63.7±8.9, mean weight 82.9±14.9 kg, and body mass index 32.2±4.8 kg/m2. The average plasma level of vWF was similar in patients with and without previous CVD, with values of 154.5±52.1 and 155.47±41.4, respectively. There was an association between diabetes mellitus (DM) and established CVD, which remained significant after adjusting for other variables included in the multivariate model. Conclusions: There was no difference in the mean plasma level of vWF among patients with MS, with and without documented CVD. The presence of DM, however, was independently associated with CVD in this population.

Avaliação do desempenho do equipamento de hematologiaSysmex XE2100D em um laboratório municipal / Evaluation of the performance of the Sysmex XE2100D hematology equipment in a municipal laboratory

Validar o analisador hematológico Sysmex XE-2100D (Kobe, Japan), descreveras etapas do processo e avaliar seu desempenho antes da sua inserção na rotina detrabalho do Laboratório Municipal de Gravataí, RS. Métodos: O estudo foi realizado com101 amostras de sangue total de pacientes adultos. Foram realizados quatro testes:precisão intraensaio, linearidade, exatidão e carreamento. Resultados: Os testes deprecisão apresentaram resultados adequados para a maioria dos parâmetros. Somentea contagem absoluta de monócitos e a quantificação de plaquetas tiveram valoressuperiores aos preconizados em algumas amostras. O coeficiente de variação foi inferiora 1,5% para a série vermelha. Para linearidade, os coeficientes de correlação foramsuperiores a 0,99; no teste de exatidão, superiores a 0,98 para a maioria dos parâmetros,e o percentual de carreamento inferior a 1,0% para todos os parâmetros analisados.Conclusão: Ao se analisarem amostras com perfis semelhantes aos testados pelofabricante, os resultados de precisão foram excelentes; entretanto, conforme esperado,as amostras com resultados alterados apresentaram coeficientes de variação maiores.Os resultados de precisão indicam a necessidade de outros estudos utilizando amostrascom resultados fora do limite de normalidade para determinação do coeficiente de variaçãoaceitável para esse tipo de amostra. Os resultados de linearidade, exatidão e carreamentoforam semelhantes aos obtidos em outros estudos. Considerando os resultados obtidos,o equipamento foi aprovado para utilização no laboratório...

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 and large laboratories, improving the quality of microscopic analysis in complete blood counts.

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 ...

Lamellar body count and stable microbubble test on tracheal aspirates from infants for the diagnosis of respiratory distress syndrome.

OBJECTIVES: To evaluate the performance of lamellar body count in tracheal aspirates from intubated preterm babies to predict respiratory distress syndrome. DESIGN: Case-control study. SETTING: Three neonatal intensive care units. PATIENTS: Seventy-two patients not older than 3 days were included in the study, 38 preterm infants with respiratory distress syndrome, 16 preterms without respiratory distress syndrome, and 18 term infants. All required mechanical ventilation. INTERVENTIONS: Lamellar body count was performed in an automated cell counter. Tracheal samples were diluted in dithiothreitol without centrifugation and kept frozen at -20°C until use. Samples were placed in a dithiothreitol-containing test tube at a ratio of one part tracheal aspirate to six parts dithiothreitol solution, vortexed for 10 secs, and aspirated by the cell counter. Lamellar body count was performed using the platelet channel. All results were multiplied by seven. The stable microbubble test was done for comparison. MEASUREMENTS: Lamellar body count and stable microbubble test. MAIN RESULTS: Lamellar body count was significantly lower in the respiratory distress syndrome group compared with the non respiratory distress syndrome preterm group and also with the term group. The median and interquartile range obtained for lamellar body count were 38,500/µL (14,000-112,000) for the respiratory distress syndrome group, 822,500/µL (442,000-962,500) for the non respiratory distress syndrome preterm group, and 633,000/µL (322,000-1,608,000) for the term group (p < .001). The sensitivity and specificity of lamellar body count and stable microbubble test for the diagnosis of respiratory distress syndrome were calculated, taking into consideration the respiratory distress syndrome and the non respiratory distress syndrome preterm groups. Considering a cutoff point of 200,000 lamellar bodies/µL, lamellar body count sensitivity was 92.1% (95% confidence interval 78.6-98.3) and lamellar body count specificity was 93.8% (95% confidence interval 69.8-99.8). The area under the curve was 0.94 (95% confidence interval 0.84-1.00). CONCLUSIONS: Lamellar body count and stable microbubble test can be rapidly and easily performed on tracheal aspirates and they seem to have very good performance for diagnosing respiratory distress syndrome in intubated patients.

Lamellar body count and stable microbubble test on gastric aspirates from preterm infants for the diagnosis of respiratory distress syndrome.

BACKGROUND: Lamellar body count (LBC) in amniotic fluid is being used to identify infants at risk of respiratory distress syndrome (RDS) who would benefit from surfactant prophylaxis or very early therapy. The test in gastric aspirates of newborns has not been properly explored. OBJECTIVE: The main objective of this research was to evaluate the performance of LBC alone or in combination with the stable microbubble test (SMT), done on gastric aspirates from preterm babies to predict RDS. METHODS: A total of 34 preterm infants with RDS and 29 without RDS, with a gestational age between 24 and 34 weeks, were included in the study. Gastric fluid was collected in the delivery room. A diluent (dithiothreitol) allowed all samples to be processed, even the thickest and non-homogeneous ones, without centrifugation. The SMT was done for comparison. RESULTS: The best cut-off value was <42,000 lamellar bodies/microl to predict RDS, with a sensitivity of 92% (95% CI 73-100%) and specificity of 86% (95% CI 77-95%). The area under the receiver-operating characteristic curve was 0.928 (95% CI 0.86-0.99). SMT showed similar results. LBC and SMT together in series (positive result if both tests were positive) showed a sensitivity of 100% and a specificity of 86%. CONCLUSION: LBC on gastric aspirates diluted in a solution of dithiothreitol can be rapidly and easily performed, and may be used alone or in combination with SMT as a predictor of RDS, allowing selective prophylaxis or very early treatment only in surfactant-deficient newborns.

Fructose-1,6-bisphosphate inhibits in vitro and ex vivo platelet aggregation induced by ADP and ameliorates coagulation alterations in experimental sepsis in rats.

Sepsis is a systemic response to an infection that leads to a generalized inflammatory reaction. There is an intimate relationship between procoagulant and proinflammatory activities, and coagulation abnormalities are common in septic patients. Pharmaceutical studies have focused to the development of substances that act on coagulation abnormalities and on the link between coagulation and inflammation. Fructose-1,6-bisphosphate (FBP) is a high-energy glycolitic metabolite that in the past two decades has been shown therapeutic effects in great number of pathological situations, including sepsis. The aims of this study were to assess the effects of FBP on platelet aggregation in vitro and ex vivo in healthy and septic rats and evaluate the use of FBP as a treatment for thrombocytopenia and coagulation abnormalities in abdominal sepsis in rat. FBP inhibited platelet aggregation (P < 0.001) in vitro in healthy rats from the smallest dose tested, 2.5 mM, in a dose-dependent manner. The mean effective dose calculated was 10.6 mM. The highest dose tested, 40 mM, completely inhibited platelet aggregation (P < 0.001) induced by ADP. Platelet aggregation in plasma from septic rats was inhibited only with higher doses of FBP, starting from 20 mM (P < 0.001). The calculated mean effective dose was 19.3 mM. Ex vivo platelet aggregation in septic rats was significantly lower (P < 0.05) than healthy rats and the treatment with FBP, at the dose of 2 g/kg, diminished the platelet aggregation at the extension of 27% (P < 0.001), suggesting that FBP is a potent platelet aggregation inhibitor in vivo. Moreover, treatment with FBP 2 g/kg prevented thrombocytopenia (P < 0.001), prolongation of prothrombin and partial thromboplastin time (P < 0.001), but not fibrinogen, in septic rats. The most important findings in this study are that FBP is a potent platelet aggregation inhibitor, in vitro and ex vivo. It presents protective effects on coagulation abnormalities, which can represent a treatment against DIC. The mechanisms for these effects remain under investigation.

Angiotensin converting enzyme ( ACE ) DD genotype: relationship with venous thrombosis

O troemboembolismo venoso (TEV) é uma doenca multifatorial associada com fatores de risco adquiridos e hereditários. Vários polimorfismos, tais como fator V de Leiden, mutacão G20210A da protrombina e as deficiências de proteína C, proteína S e anti-trombina são considerados fatores de risco para TEV. A enzima conversora da angiotensina (ECA) afeta a hemostasia diminuindo a fibrinólise. O polimorfismo no gene da ECA, caracterizado pela insercão/delecão de um fragmento de 287 pb no intron16, está relacionado a variacões nos níveis séricos da enzima. O genótipo DD foi associado com aumento de risco para TEV. Este estudo examinou a freqüência dos alelos I e D e a sua associacão com trombose venosa em um grupo de indivíduos do Sul do Brasil. Foram analisados 71 pacientes com trombose venosa profunda e/ou tromboembolismo pulmonar e 71 indivíduos sem história de trombose. A genotipagem foi realizada através da reacão em cadeia da polimerase. As freqüências do alelo D e do genótipo DD foram, respectivamente, 51,4% e 22,5% para os pacientes, e 64,7% e 45,0% para os controles. A razão de chance (odds ratio = OR) para a hipótese dominante (genótipos DD+ID versus genótipo II) foi 0,75 (IC 95%; 0,29-1,93) e a OR para a hipótese recessiva (genótipo DD versus genótipos ID+II) foi 0,35 (IC 95%; 0,16-0,78). Concluindo, nossos resultados sugerem que o genótipo DD não representa um fator de risco para TEV e pode exercer um efeito protetor para trombose venosa.