The homing of bone marrow stem cells is differentially activated in ischemic and valvular heart diseases and influenced by beta-blockers.

BACKGROUND: Cell homing is the mechanism by which an injury releases signaling molecules that cause recruitment, proliferation, migration and differentiation of progenitor cells. Stromal derived factor-1 (SDF-1) and its receptor CXCR4 are key molecules involved in homing and little is known about their activation in cardiopathies. Here, we assessed the homing activation status of bone marrow cells (BMC) concerning the SDF-1 and CXCR4 expression in ischemic (IHD) and valvular (VHD) heart diseases. METHODS: The SDF-1 and inflammatory profile were analyzed by ELISA from plasma obtained bone marrow of ischemic heart patients (IHD, n = 41), valvular heart patients (VHD, n = 30) and healthy controls (C, n = 9). Flow cytometry was used to evaluate CXCR4 (CD184) expression on the surface of bone marrow cells, and the CXCR4 expression was estimated by real-time quantitative PCR. RESULTS: The SDF-1 levels in the groups IHD, VHD and control were, respectively, 230, 530 and 620 pg/mL (P = 0.483), and was decreased in VHD patients using beta-blockers (263 pg/mL) when compared with other (844 pg/mL) (P = 0.023). Compared with IHD, the VHD group showed higher CXCR4 (P = 0.071) and CXCR7 (P = 0.082) mRNA expression although no difference in the level of CXCR4+ bone marrow cells was found between groups (P = 0.360). CONCLUSION: In conclusion, pathophysiological differences between IHD and VHD can affect the molecules involved in the activation of homing. In addition, the use of beta-blockers appears to interfere in this mechanism, a fact that should be considered in protocols that use BMC.

Mesenchymal stem cells from sternum: the type of heart disease, ischemic or valvular, does not influence the cell culture establishment and growth kinetics.

BACKGROUND: In an attempt to increase the therapeutic potential for myocardial regeneration, there is a quest for new cell sources and types for cell therapy protocols. The pathophysiology of heart diseases may affect cellular characteristics and therapeutic results. METHODS: To study the proliferative and differentiation potential of mesenchymal stem cells (MSC), isolated from bone marrow (BM) of sternum, we made a comparative analysis between samples of patients with ischemic (IHD) or non-ischemic valvular (VHD) heart diseases. We included patients with IHD (n = 42) or VHD (n = 20), with average age of 60 years and no differences in cardiovascular risk factors. BM samples were collected (16.4 ± 6 mL) and submitted to centrifugation with Ficoll-Paque, yielding 4.5 ± 1.5 × 107 cells/mL. RESULTS: Morphology, immunophenotype and differentiation ability had proven that the cultivated sternal BM cells had MSC features. The colony forming unit-fibroblast (CFU-F) frequency was similar between groups (p = 0.510), but VHD samples showed positive correlation to plated cells vs. CFU-F number (r = 0.499, p = 0.049). The MSC culture was established in 29% of collected samples, achieved passage 9, without significant difference in expansion kinetics between groups (p > 0.05). Dyslipidemia and the use of statins was associated with culture establishment for IHD patients (p = 0.049 and p = 0.006, respectively). CONCLUSIONS: Together, these results show that the sternum bone can be used as a source for MSC isolation, and that ischemic or valvular diseases do not influence the cellular yield, culture establishment or in vitro growth kinetics.

Combined Analysis of Endothelial, Hematopoietic, and Mesenchymal Stem Cell Compartments Shows Simultaneous but Independent Effects of Age and Heart Disease.

Clinical trials using stem cell therapy for heart diseases have not reproduced the initial positive results obtained with animal models. This might be explained by a decreased regenerative capacity of stem cells collected from the patients. This work aimed at the simultaneous investigation of endothelial stem/progenitor cells (EPCs), mesenchymal stem/progenitor cells (MSCs), and hematopoietic stem/progenitor cells (HSCs) in sternal bone marrow samples of patients with ischemic or valvular heart disease, using flow cytometry and colony assays. The study included 36 patients referred for coronary artery bypass grafting or valve replacement surgery. A decreased frequency of stem cells was observed in both groups of patients. Left ventricular dysfunction, diabetes, and intermediate risk in EuroSCORE and SYNTAX score were associated with lower EPCs frequency, and the use of aspirin and ß-blockers correlated with a higher frequency of HSCs and EPCs, respectively. Most importantly, the distribution of frequencies in the three stem cell compartments showed independent patterns. The combined investigation of the three stem cell compartments in patients with cardiovascular diseases showed that they are independently affected by the disease, suggesting the investigation of prognostic factors that may be used to determine when autologous stem cells may be used in cell therapy.

Comparação das metodologias óptica e de impedância para a determinação de parâmetros hematológicos no analisador Sysmex-XN

Objetivo: Comparar as metodologias de impedância e óptica para contagem de eritrócitos (RBC) e dosagem de hemoglobina (HGB) como forma de corrigir a contagem de eritrócitos em casos de pacientes que apresentam índice de concentração de hemoglobina corpus- cular média (CHCM) acima do ponto de corte. Métodos: Para comparar as metodologias, foram utilizadas amostras com CHCM normal e aumentado. Os parâmetros avaliados foram RBC, HGB e CHCM. Resultados: As amostras com valores de CHCM aumentado demonstraram uma correlação significativa entre RBC e HGB, porém o CHCM apresen- tou uma correlação fraca. Observamos que a metodologia por impedância obteve maior sucesso na correção do CHCM quando as amostras apresentavam aglutinação. Conclusão: A metodologia óptica é uma boa alternativa para correção de valores aumentados de CHCM em situações específicas, mas para casos de aglutinação não se mostrou efetiva.

Objetctive: Compare the impedance and optical methodologies for red blood cells (RBC) count and hemoglobin (HGB) measurement as a way to correct the RBC count in cases of patients who have a MCHC index above the cutoff. Methods: To compare the methodologies, samples with normal and augmented MCHC were used. The parameters evaluated were RBC, HGB and MCHC. Results: Samples with increased MCHC values demonstrated a significant correlation between RBC and HGB, but the MCHC showed a weak correlation. We observed that the impedance methodology was more successful in correcting the MCHC when the samples presented agglutination. Conclusion: Optical methodology is a good alternative for correction of increased CHCM values in specific situations, but for cases of agglutination it has not been shown to be effective.

Inclusões semelhantes aos corpúsculos de Russell em blastos mieloides / Inclusions similar to Russell bodies in myeloblasts

Alterações nos mecanismos celulares resultam no aparecimento de corpúsculos de Russel, que são inclusões intracitoplasmáticas basofílicas. Estas inclusões ocorrem devido a uma indigestão celular de proteínas e podem ser observadas em diversas condições patológicas como leucemias, mieloma múltiplo, linfoma de Burkitt e gastrite com corpúsculos de Russell, sendo raramente observadas em leucemias mieloides agudas. O presente estudo relata a presença de inclusões semelhantes aos corpúsculos de Russell em blastos no sangue periférico de uma paciente de 51 anos com leucemia mieloide aguda.

Changes in cell mechanisms result in the appearance of Russel bodies, which are basophilic intracytoplasmic inclusions. These inclusions occur due to cellular indigestion of proteins, and can be observed in several pathological conditions such as leukemias, multiple myeloma, Burkitt's lymphoma, and gastritis with Russell bodies, but rarely observed in acute myeloid leukemias. The present study reports the presence of inclusions similar to Russell bodies in peripheral blood blasts in a 51-year-old female patient with acute myeloid leukemia.

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.

Serum bilirubin concentration in patients with an established coronary artery disease.

Bilirubin has been considered an antioxidant, with capacity to remove reactive species of oxygen. Studies have suggested that an increased bilirubin level promotes protection against atherosclerosis. The case group was composed of 100 patients with coronary artery disease and the control group 100 patients with normal coronaries. Blood samples were collected to determine bilirubin concentrations. Bivariate analysis, multiple logistic regression models, and Spearman's correlation index were performed. A P value < 0.05 was considered to be significant. The case group was predominantly composed of men and the control group of women, with a mean age of 60 +/- 8.8 versus 56 +/- 10.9 (P = 0.015). The total bilirubin average was significantly higher in the control group than in the case group (0.76 mg/dL versus 0.39 mg/dL, P < 0.001). The level of ultrasensitive C reactive protein (us-CRP) was increased in the case group (3.63 mg/L versus 0.93 mg/L, P < 0.001). Although the correlation index for this inverse association has been weak, both are independently associated with a higher prevalence of coronary artery disease, total bilirubin 3 mg/L (OR: 1.17; IC: 1.04-1.33; P = 0.009). Reduced serum levels of bilirubin were shown to be associated with a higher prevalence of coronary artery disease emerging as a new potential risk factor marker. Additional studies are still necessary to confirm and demonstrate the association of these findings with clinical outcomes.

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 ...

Perfil lipídico de transplantados renais em uso de terapia imunossupressora / Lipid profile in post renal transplant patients treated with immunosuppressive therapy

Objetivo: A prevalência de alterações no perfil lipídico dos transplantados renais é muito alta, sendo a doença cardiovascular uma das principais causas de mortalidade nesses pacientes. Mesmo após o transplante renal, as complicações cardíacas não diminuem. O objetivo foi avaliar o perfil lipídico de pacientes transplantados renais em uso de terapia imunossupressora e relacionar qual dos imunossupressores estaria associado a alterações no perfil lipídico. Métodos: Estudo retrospectivo realizado com 186 pacientes em terapia imunossupressora pós-transplante renal com ciclosporina, tacrolimus ou sirolimus. Foram coletadas amostras dos pacientes que realizaram transplante renal entre 2002 e 2007 , na Irmandade Santa Casa de Misericórdia de Porto Alegre, e que foram incluídos na pesquisa. Resultados: Foi observado que os níveis de colesterol aumentaram nos pacientes em uso de ciclosporina e sirolimus. O HDL-C mostrou-se mais elevado em todos os grupos, independente do tempo, após o transplante renal. Quanto ao LDL-C, verificou-se diminuição com o passar do tempo de tratamento no grupo que fazia uso de ciclosporina. A razão colesterol total/HDL-C diminuiu com o tratamento de ciclosporina e de tacrolimus após 24 meses. Conclusão: Os resultados sugerem que o tacrolimus e a ciclosporina são os imunossupressores menos prejudiciais ao perfil lipídico dos pacientes transplantados renais, no entanto, o sirolimus está associado à maior alteração no perfil lipídico, podendo levar os pacientes a um pior prognóstico cardíaco.

Associação da concentração de bilirrubina sérica com a carga aterosclerótica em pacientes com doença arterial coronariana estabelecida / Association of serum bilirubin concentration with atherosclerotic burden in patients with established coronary artery disease

Fundamentos: As doenças cardiovasculares são um importante problema de saúde pública, e as respostas adaptativas e protetoras da vasculatura contra o estresse oxidativo são importantes na prevenção da aterosclerose. A bilirrubina tem sido considerada um antioxidante, com capacidade de remover espécies reativas de oxigênio. Estudos têm demonstrado que a concentração de bilirrubina sérica aumentada promove proteção contra aterosclerose. Objetivo: Avaliar a possível existência de relação entre a carga aterosclerótica de pacientes com doença arterial coronariana e a concentração de bilirrubina. Método: Estudo transversal composto por 81 pacientes com diagnóstico angiográfico estabelecido de doença arterial coronariana, submetidos a revascularização miocárdica. Foram avaliados os fatores de risco clássicos e a dosagem de bilirrubina. Foram utilizados os testes do qui-quadrado, teste t de Student ou Mann-Whitney, sendo considerado p < 0,05 estatisticamente significativo. Resultados: A amostra foi composta predominantemente por homens, com média de idade de 59,32 ± 9,28 anos no grupo com lesão mínima/moderada e 61,79 ± 7,55 anos no grupo com lesão grave. A mediana da bilirrubina total no grupo com lesão mínima/moderada foi de 0,34 mg/dL, enquanto no grupo com lesão grave foi de 0,43 mg/dL, diferente do esperado. A concentração de bilirrubina total foi maior no grupo com carga aterosclerótica grave que no grupo com carga mínima/moderada, embora sem significância estatística (p = 0,241). Conclusão: Níveis séricos de bilirrubina não se mostraram úteis na detecção ou predição da gravidade da lesão aterosclerótica dos pacientes com doença arterial coronariana.

Background: Cardiovascular diseases are an important public health problem. Adaptive and protective responses of the vascular system against oxidative stress are important in the prevention of atherosclerosis. Bilirubin has been considered an antioxidant, with the capacity to remove reactive species of oxygen. Previous studies have demonstrated that increased serum bilirubin concentrations promote protection against atherosclerosis. Purpose: To evaluate the relationship between the atherosclerotic burden and the bilirubin concentration of patients with coronary artery disease. Method: Transversal study composed of 81 patients with established angiographic diagnosis of coronary artery disease, submitted to myocardial revascularization. Classical risk factors and bilirubin levels were assessed. The chisquare, the t Student test or the Mann-Whitney test were used with p value significant at < 0.05. Results: The sample was composed predominantly of men, with a mean age of 59.32 ± 9.28 years in the minimum/moderate atherosclerotic burden group and 61.79 ± 7.55 years in the severe burden group. The median of total bilirubin in the minimum/moderate atherosclerosis burden group was 0.34 mg/dL and 0.43 mg/dL in the severe burden group. Differently than expected, the concentration of total bilirubin was higher in the group with severe atherosclerotic burden than in the group with minimum/moderate burden, although without statistical significance (p = 0.241). Conclusion: Serum bilirubin levels were not useful in the detection or prediction of the severity of atherosclerotic lesions in patients with coronary artery disease.