Everolimus-eluting bioresorbable scaffolds and metallic stents in diabetic patients: a patient-level pooled analysis of the prospective ABSORB DM Benelux Study, TWENTE and DUTCH PEERS.

BACKGROUND: Several studies compared everolimus-eluting bioresorbable scaffolds (EE-BRS) with everolimus-eluting stents (EES), but only few assessed these devices in patients with diabetes mellitus. AIM: To evaluate the safety and efficacy outcomes of all-comer patients with diabetes mellitus up to 2 years after treatment with EE-BRS or EES. METHODS: We performed a post hoc pooled analysis of patient-level data in diabetic patients who were treated with EE-BRS or EES in 3 prospective clinical trials: The ABSORB DM Benelux Study (NTR5447), TWENTE (NTR1256/NCT01066650) and DUTCH PEERS (NTR2413/NCT01331707). Primary endpoint of the analysis was target lesion failure (TLF): a composite of cardiac death, target vessel myocardial infarction or clinically driven target lesion revascularization. Secondary endpoints included major adverse cardiac events (MACE): a composite of all-cause death, any myocardial infarction or clinically driven target vessel revascularization, as well as definite or probable device thrombosis (ST). RESULTS: A total of 499 diabetic patients were assessed, of whom 150 received EE-BRS and 249 received EES. Total available follow-up was 222.6 patient years (PY) in the EE-BRS and 464.9 PY in the EES group. The adverse events rates were similar in both treatment groups for TLF (7.2 vs. 5.2 events per 100 PY, p = 0.39; adjusted hazard ratio (HR) = 1.48 (95% confidence interval (CI): 0.77-2.87), p = 0.24), MACE (9.1 vs. 8.3 per 100 PY, p = 0.83; adjusted HR = 1.23 (95% CI: 0.70-2.17), p = 0.47), and ST (0.9 vs. 0.6 per 100 PY, p > 0.99). CONCLUSION: In this patient-level pooled analysis of patients with diabetes mellitus from 3 clinical trials, EE-BRS showed clinical outcomes that were quite similar to EES.

The role of D2 dopamine receptors in oxytocin induced place preference and anxiolytic effect.

Neuropeptide oxytocin (OT) is involved in the regulation of social and non-social behaviour. The central nucleus of amygdala (CeA), part of the limbic system, plays an important role in learning, memory, anxiety and reinforcing mechanisms. CeA has been shown to be rich in OT receptors in rodents. Our previous findings indicated that OT in the rat CeA has a dose dependent rewarding and anxiolytic effect. The aim of our present study was to examine in the CeA the possible interaction of OT and D2 dopamine (DA) receptor antagonist Sulpiride on reinforcement in place preference test and on anxiety in elevated plus maze test. Wistar rats were microinjected bilaterally with 10 ng OT. In different group of animals 4 µg D2 DA receptor antagonist was applied. Other animals received D2 DA receptor antagonist 15 min before 10 ng OT treatment or vehicle solution into the CeA. Rats receiving 10 ng OT spent significantly longer time in the treatment quadrant during the test session in conditioned place preference test. Prior treatment with D2 DA receptor antagonist blocked the rewarding effects of OT. Antagonist in itself did not influence the time rats spent in the treatment quadrant. In elevated plus maze test, rats receiving 10 ng OT spent significantly longer time on the open arms. Prior treatment with D2 DA receptor antagonist blocked the effects of OT. Our results show that DA system plays a role in positive reinforcing and anxiolytic effects of OT because D2 DA receptor antagonist can block these actions.

Everolimus-eluting bioresorbable scaffolds for treatment of coronary artery disease in patients with diabetes mellitus: the midterm follow-up of the prospective ABSORB DM Benelux study.

BACKGROUND: Percutaneous coronary intervention (PCI) in patients with diabetes mellitus (DM) remains challenging even with modern drug-eluting stents (DES) due to high rates of repeat revascularization. Everolimus-eluting bioresorbable scaffolds (EE-BRS) might allow for repeat intervention prolonging the time interval of percutaneous treatment options. METHODS: The ABSORB DM Benelux Study is a dedicated prospective, international study to evaluate the midterm safety and efficacy of EE-BRS in DM patients. All DM patients that received ≥ 1 EE-BRS for any indication were enrolled and prospectively followed. Study endpoints were major adverse cardiac events (MACE): a composite of all-cause death, any myocardial infarction (MI) and ischemic-driven target vessel revascularization (TVR); target lesion failure (TLF): a composite of cardiac death (CD), target vessel MI, and ischemic-driven target lesion revascularization (TLR), as well as definite or probable scaffold thrombosis (ScT). RESULTS: Between April 2015 till March 2017, 150 DM patients and 188 lesions were treated and followed up to 3 years. Device implantation success was 100%. MACE occurred in 15.2% (event rate of 8.8 per 100 PY). TLF was reported in 11.7% (7.0 events per 100 PY). CD, target vessel MI, ischemic-driven TLR occurred in 3.4%, 3.6% and 5.5% respectively, while ScT was observed in 1.4%. There were no occurrences of late or very late ScT. CONCLUSION: EE-BRS treatment in DM patients shows comparable midterm safety and efficacy outcomes when historically compared with modern DES. New-generation EE-BRS might offer an attractive alternative to metallic DES in treatment of fast progressing atherosclerosis population as in DM patients. Trial registration NTR5447. Registered 05 October 2015, retrospectively registered.

The 1­year safety and efficacy outcomes of Absorb bioresorbable vascular scaffolds for coronary artery disease treatment in diabetes mellitus patients: the ABSORB DM Benelux study.

BACKGROUND: Diabetes mellitus (DM) patients show higher rates of repeat revascularisation even in the era of modern drug-eluting stents (DES). The concept of bioresorbable scaffolds is becoming captivating, as it might allow for repeat interventions, prolonging the time span during which patients can be treated by percutaneous coronary intervention (PCI). AIMS: We intend to evaluate the short- and long-term safety and efficacy of Absorb bioresorbable vascular scaffolds (Absorb BVS) in the treatment of coronary artery disease (CAD) in DM patients for any indication. METHODS: The ABSORB DM Benelux is an international prospective study in DM patients who have undergone PCI with ≥1 Absorb BVS. Major adverse cardiac events (MACE) at 1 year was the primary endpoint, defined as a composite of all-cause death, any myocardial infarction (MI) and ischaemia-driven target vessel revascularisation (TVR). Secondary endpoints were target lesion failure (TLF) and definite or probable scaffold thrombosis (ScT). RESULTS: Between April 2015 and March 2017, 150 DM patients and 188 non-complex lesions were treated. Device implantation was successful in 100%. MACE occurred in 14 (9.5%) patients, with all-cause death occurring in 4 (2.7%), any MI in 6 (4.1%) and ischaemia-driven TVR in 7 (4.8%) respectively. TLF was reported in 11 (7.5%). Definite and probable ScT was observed in 2 (1.4%). CONCLUSION: Absorb BVS for treatment of anatomically low-risk patients with DM show acceptable safety and efficacy outcomes at 1 year. If these promising results are confirmed after a longer follow-up period, new-generation bioresorbable scaffolds combined with refinement of implantation techniques might open new horizons for CAD treatment in DM patients.

Transradial and transulnar access for iliac artery interventions using sheathless guiding systems: A feasibility study.

PURPOSE: Our aim was to evaluate the acute success and complication rates of the transradial and transulnar access for iliac artery stenting using sheathless guiding systems. METHODS: Clinical and angiographic data from 156 consecutive patients with symptomatic iliac artery stenosis who were treated with transradial or transulnar access were evaluated. All patients underwent Duplex ultrasound before and after the intervention. The primary endpoints were the procedural success rate, major adverse events, and access site complication rates. The secondary endpoints were the angiographic result of the iliac artery intervention, fluoroscopy time, X-ray dose, procedure length, crossover rate to another puncture site and hospitalization duration. The impact of the learning curve was also investigated, along with right or left radial access. RESULTS: The indication for the intervention was intermittent claudication in 109 patients (69.9%), critical limb ischemia in 44 (28.2%) subjects and acute limb ischemia in three individuals (1.9%). Technical success was achieved in 155 patients (99.4%), with a crossover rate of 3.8%. Radial and ulnar artery access was used in 151 (96.8%) and 7 (4.5%) patients, respectively. The Ankle-brachial index increased from 0.69 [0.65-0.72] to 0.91 [0.88-0.95] as a result of the procedures (P < 0.001). The cumulative incidence of major adverse events was 3.8% at the 2-month follow-up (0% in patients with intermittent claudication and 13.8% in patients with critical limb ischemia). Radial artery access site complications were encountered in eight patients (5.1%). We documented decreased X-ray doses (1742.0 [783.9-2701] vs. 1435 [991.1-1879] vs. 692.8 [275.3-1110] Gy cm-2 P < 0.05) over time; however, the fluoroscopy time, procedure time, and contrast consumption were not significantly different. Left hand access was not associated with significantly better results than right radial artery access. CONCLUSIONS: Iliac artery stenting can be safely and effectively performed using radial or ulnar artery access and sheathless guiding catheters, with acceptable complication rates and high levels of technical success. The physician learning curve plays an important role in decreasing the X-ray dose. © 2016 The Authors. Catheterization and Cardiovascular Interventions Published by Wiley Periodicals, Inc.

Autologous bone marrow stem cell neurotransplantation in stroke patients. An open study.

PURPOSE: Bone marrow stem cells (BMSC) were transplanted into the perilesional area in five patients bearing sequels of stroke, to evaluate the safety of the procedure and tolerance to the transplanted cells. METHODS: Cells were obtained from bone marrow samples taken from the same patient and stereotactically implanted into the targets, determined using a combination of images, and trans-operative recording of multiunit activity. The cells were implanted in several points along tracts in the perilesional region. RESULTS: No important adverse events derived from surgery or transplant were observed during the one year follow-up period, or detected using a combination of tests and functional measurements applied pre- and post-surgically. In contrast, some improvements were observed regarding the neurological condition of the patients, but the small number of patients in the study does not allow any conclusive statement. CONCLUSIONS: Our results demonstrate that BMSC can be safely transplanted into the brain of patients, with excellent tolerance and without complications, using the methods described here.

Cross-cultural adaptation of the disabilities of the arm, shoulder, and hand (DASH) questionnaire into Hungarian and investigation of its validity in patients with systemic sclerosis.

OBJECTIVE: To adapt and validate the Hungarian version of the DASH and the shorter QuickDASH Outcome Measures and to establish their validity in patients with systemic sclerosis (SSc). METHODS: The Hungarian adaptation of the questionnaires was performed using forward/backward translations, expert and lay reviews. 128 patients completed the DASH, the Health Assessment Questionnaire (HAQ-DI), the Modified HAQ-DI for patients with SSc (SSc-HAQ), and the Short Form Health Survey (SF-36) questionnaire. 76 patients participated in a 12-month follow-up examination. Sensitivity to change was estimated using the standardized response mean (SRM). RESULTS: Cronbach alpha in the DASH sections were between 0.94-0.97. The intraclass correlation coefficient for the test-retest reliability of DASH was 0.89. DASH scores showed a correlation with both SSc-HAQ and the physical dimensions of the SF-36 (Spearman's rho: 0.89, -0.77 and -0.42, respectively). The SRM of DASH was 0.64 among the scleroderma patients with worsening HAQ-DI status. CONCLUSIONS: The Hungarian version of the DASH and QuickDASH demonstrated equivalent reproducibility, internal consistency and validity to the originals. The strong correlations of the DASH and QuickDASH with the HAQ-DI, and with the physical dimensions of the SF-36 show that the disability of the patient with SSc is predominantly caused by the functional impairment of the upper limb. Because both questionnaires were valuable tools for measuring upper extremity function and joint damage in SSc patients, we recommend the shorter and simpler QuickDASH for everyday clinical use.

Alterations of conditioned taste aversion after microiontophoretically applied neurotoxins in the medial prefrontal cortex of the rat.

The prefrontal cortex (PFC) has been reported to be essential in neural control of feeding. In the present study, we aimed to provide a complex characterization of behavioral consequences of PFC microlesions in CFY rats. Kainic acid (KA) was microiontophoretically applied into the mediodorsal division of PFC to damage intrinsic neurons, whereas in another group of rats, 6-hydroxydopamine (6-OHDA) was microiontophoretized into the same region to destroy catecholaminergic (CA) projection fiber terminals. Body weights, food and fluid intake of both lesioned and (sham-operated or intact) control animals were daily measured. Effects of intracellular dehydration and water deprivation were also studied. Open field activity, stereotyped behaviors, and orientation towards visual and somesthetic stimuli were pre- and postoperatively tested. To examine hypothesized consequences of mPFC microlesions on central taste information processing, the acquisition and retention of saccharine conditioned taste aversion (CTA) were studied. No major changes were recorded in body weights, food and water consumption. Dehydration or deprivation similarly increased water intake in all animals. Scores of open field activity and stereotyped behaviors in the 6-OHDA group were significantly higher than those of the other groups. As the main findings of the present studies, both KA and 6-OHDA lesioned rats displayed significant deficits in CTA acquisition and retention tests. These results suggest that the medial PFC has a substantial role in both the formation and the retrieval of CTA. Furthermore, the present findings also indicate the general significance of prefrontal CA mechanisms in the organization of goal-directed, adaptive behaviors.

Aislamiento y caracterización de células mesenquimales derivadas de tejido adiposo / Isolation and characterization of mesenchymal cells from adipose tissue

Introducción : Existe un creciente interés científico en el potencial terapéutico de las células madre mesenquimales derivadas de tejido adiposo ( ADSCs, en inglés). Estas células son abundantes en el tejido adiposo, son de fácil obtención y con un alto potencial de diferenciación hacia linajes celulares especializados incluyendo adipocitos, osteocitos, condrocitos, miocitos, cardiomiocitos, tenocitos, vasos sanguíneos y neuronas. Este trabajo se desarrolló con el objetivo de implementar en el laboratorio un procedimiento para aislar y cultivar ADSCs, con características que corresponden a las informadas para este linaje celular.Método: los precursores de células adiposas humanas se obtuvieron de tejido subcutáneo abdominal. Las células se separaron enzimáticamente del tejido y se decantaron por centrifugación, luego de cultivadas, se caracterizaron en su capacidad de diferenciación y por su marcadores fenotípicos.Resultados: Las ADSCs aisladas se replicaron en estas condiciones de cultivo y mantuvieron un fenotipo estable durante todo el período de estudio. Se comprobó su potencial adipogénico y osteogénico in vitro, como corresponde a las células madre mesenquimales. El estudio por citometría de flujo mostró que estas células expresan CD73, CD90 y CD105 y son negativas para los marcadores de linaje hematopoyético CD34 y CD45.En los ensayos de inhibición in vitro, las ADSCs demostraron su capacidad para inhibir la proliferación de células T humanas. Conclusiones : La caracterización fenotípica y funcional de las ADSCs obtenidas a partir del tejido adiposo abdominal demuestra que es posible la obtención mediante cultivo in vitro de células mesenquimales humanas sin inducir diferenciación espontánea, manteniendo su integridad funcional y altos niveles de proliferación, lo que sienta las bases para el inicio de ensayos preclínicos y su uso futuro en la terapia celular en nuestro país(AU)

Introduction : There is growing scientific interest in the therapeutic potential of stem cells derived from adipose tissue (ADSCs). These cells are abundant in adipose tissue, are readily available and have a high potential fordifferentiation into specialized cell lineages including adipocytes, osteocytes, chondrocytes, myocytes, cardiomyocytes, tenocyte, endothelial cells and neurons. The aim of this work was to isolate, cultivate andcharacterize ADSCs.Methods : human adipose precursor cells were obtained from abdominal subcutaneous tissue. Cells were enzymatically separated from the tissue and decanted by centrifugation, cultured and finally analyzed.Results : The ADSCs were able to replicate in our culture conditions. The cells maintained their phenotype in different passages throughout the study period, confirming its feasibility for in vitro culture. Also the ADSCs were induced to adipogenic and osteogenic differentiation, verifying its potential as mesenchymal stem cells in vitro. The flow cytometric study showed that these cells expressed CD73, CD90 and CD105 (markers of mesenchymal cells) and they were negative for CD34 and CD45 (hematopoieticcell markers). The ADSCs were able to inhibit in vitro the proliferation of T cells.Conclusions : It is possible to obtain ADSCs by in vitro cultivation without adipogenic induction, maintaining its functional integrity and high proliferation; this cell could be an important tool for the cellular therapy in our country(AU)

Aislamiento y caracterización de células mesenquimales derivadas de tejido adiposo / Isolation and characterization of mesenchymal cells from adipose tissue

Introducción : Existe un creciente interés científico en el potencial terapéutico de las células madre mesenquimales derivadas de tejido adiposo ( ADSCs, en inglés). Estas células son abundantes en el tejido adiposo, son de fácil obtención y con un alto potencial de diferenciación hacia linajes celulares especializados incluyendo adipocitos, osteocitos, condrocitos, miocitos, cardiomiocitos, tenocitos, vasos sanguíneos y neuronas. Este trabajo se desarrolló con el objetivo de implementar en el laboratorio un procedimiento para aislar y cultivar ADSCs, con características que corresponden a las informadas para este linaje celular. Método: los precursores de células adiposas humanas se obtuvieron de tejido subcutáneo abdominal. Las células se separaron enzimáticamente del tejido y se decantaron por centrifugación, luego de cultivadas, se caracterizaron en su capacidad de diferenciación y por su marcadores fenotípicos. Resultados: Las ADSCs aisladas se replicaron en estas condiciones de cultivo y mantuvieron un fenotipo estable durante todo el período de estudio. Se comprobó su potencial adipogénico y osteogénico in vitro, como corresponde a las células madre mesenquimales. El estudio por citometría de flujo mostró que estas células expresan CD73, CD90 y CD105 y son negativas para los marcadores de linaje hematopoyético CD34 y CD45.En los ensayos de inhibición in vitro, las ADSCs demostraron su capacidad para inhibir la proliferación de células T humanas. Conclusiones : La caracterización fenotípica y funcional de las ADSCs obtenidas a partir del tejido adiposo abdominal demuestra que es posible la obtención mediante cultivo in vitro de células mesenquimales humanas sin inducir diferenciación espontánea, manteniendo su integridad funcional y altos niveles de proliferación, lo que sienta las bases para el inicio de ensayos preclínicos y su uso futuro en la terapia celular en nuestro país(AU)

Introduction : There is growing scientific interest in the therapeutic potential of stem cells derived from adipose tissue (ADSCs). These cells are abundant in adipose tissue, are readily available and have a high potential fordifferentiation into specialized cell lineages including adipocytes, osteocytes, chondrocytes, myocytes, cardiomyocytes, tenocyte, endothelial cells and neurons. The aim of this work was to isolate, cultivate andcharacterize ADSCs. Methods : human adipose precursor cells were obtained from abdominal subcutaneous tissue. Cells were enzymatically separated from the tissue and decanted by centrifugation, cultured and finally analyzed. Results : The ADSCs were able to replicate in our culture conditions. The cells maintained their phenotype in different passages throughout the study period, confirming its feasibility for in vitro culture. Also the ADSCs were induced to adipogenic and osteogenic differentiation, verifying its potential as mesenchymal stem cells in vitro. The flow cytometric study showed that these cells expressed CD73, CD90 and CD105 (markers of mesenchymal cells) and they were negative for CD34 and CD45 (hematopoieticcell markers). The ADSCs were able to inhibit in vitro the proliferation of T cells. Conclusions : It is possible to obtain ADSCs by in vitro cultivation without adipogenic induction, maintaining its functional integrity and high proliferation; this cell could be an important tool for the cellular therapy in our country(AU)

Caracterización fenotípica de células madre mesenquimales humanas de médula ósea y tejido adiposo. Resultados preliminares / Phenotypic characterization of mesenchymal human stem cell from bone marrow and adipose tissue. preliminary results

Introducción:las células madre mesenquimales (CMM) poseen características fenotípicas y funcionales que les confieren un amplio potencial terapéutico por su posible uso en la terapia celular regenerativa, en el rechazo del trasplante alogénico y en enfermedades inflamatorias crónicas. Objetivo: evaluar la expresión de moléculas de membrana que permiten identificar la expresión de patrones moleculares característicos de CMM humanas mantenidas en cultivo. Métodos: se estudió la expresión fenotípica de células mononucleares procedentes de médula ósea obtenidas mediante aspiración medular, separadas por gradiente de Ficoll y cultivadas ex vivo entre los pases o subcultivos 3 y 16; y adipocitos cultivados procedentes de la extracción enzimática de tejido adiposo de donantes sanos. Se realizó doble marcaje para las moléculas CD34/CD45, CD34/CD90, CD34/CD117, y CD34/CD44. Resultados: en los resultados preliminares obtenidos se observó que las células cultivadas procedentes de médula ósea, entre los pases 4 y 8 de cultivo expresaron 45,13 por ciento de células CD34-/CD45- (doblemente negativas), lo que correspondió con el 25,24 por ciento de células CD34-/CD90+ y el 96,90 por ciento de CD34-/CD117-. En las células procedentes de cultivo de adipocitos se observó el 52,3 por ciento de CD34-/CD45- (doblemente negativas),12,31 por ciento de CD34-/CD90+,43,31 por ciento de CD34-/CD117- y 64,68 por ciento de CD34-/CD44+.Estos resultados sugieren que ambos cultivos se diferenciaron a CMM. Las CMM procedentes de adipocitos mostraron el 64,68 por ciento de células con expresión de la molécula de adhesión CD44 a la que se atribuyen propiedades funcionalescomo el asentamiento tisular. Conclusiones: estos resultados preliminares permiten corroborar que ambos métodos experimentales de cultivo son efectivos para la obtención de CMM con fines terapéuticos(AU)

Introduction:mesenchymal stem cells (MSCs) have phenotypic and functional characteristics whichgives them a broad therapeutic potential for possible use in regenerative cell therapy, allogeneic transplant rejection and chronic inflammatory diseases. Objective: to evaluate the expression of moleculemembranes expression to identify molecular patterns characteristic of human MSCs maintained in culture. Methods:the phenotypic expression of mononuclear cells from bone marrow wereobtained by bone marrow aspiration, separated by Ficoll and cultured ex vivo between passages or subcultures 3 and 16 and adipocytes cultured obtained from enzyme extraction of adipose tissue of healthy donor. Double staining was performed for molecules CD34/CD45, CD34/CD90, CD34/CD117 and CD34/CD44. Results:preliminary results showed that cultured mononuclear cells from bone marrow between passage 4 and 8 of culture expressed 45,13 percent CD34-/CD45- cells (double-negative), corresponding to 25,24 percent CD34-/CD90+ cells and 96,90 percent of CD34-/CD117-. Adipocytes from culture cells showed 52,3 percent CD34-/CD45- (double-negative), 12,31 percent cells CD34-/CD90+, 43,31 percent CD34-/CD117- (double-negative). Our results suggest that both cultures were differentiated to MSCs. Adipocytes from MSCs showed 64,68 percent of cells with expression of CD44 adhesion molecule conferring functional homing properties Conclusions:these preliminary results corroborate that the experimental methods used in cultivation are effective for obtaining MSCs with therapeutic purposes(AU)

Caracterización inmunofenotípica de pacientes con leucemia mieloide aguda / Immunophenotypic characterization of patients with acute myeloid leukemia

Introducción: la leucemia mieloide aguda incluye un grupo heterogéneo de neoplasias caracterizadas por una expansión clonal de mieloblastos, cuya clasificación involucra varios criterios, incluidos los inmunológicos. Objetivo: caracterizar el inmunofenotipo de los pacientes con leucemia mieloide aguda evaluados en el Instituto de Hematología e Inmunología. Métodos: se realizó un estudio descriptivo transversal de los pacientes diagnosticados con este tipo de leucemia, cuyas muestras de sangre fueron procesadas en el Departamento de Inmunología en el período 2008-2012. Se usó un ultramicrométodo inmunocitoquímico que utiliza un panel de anticuerpos monoclonales específicos de antígenos mieloides y linfoides. Las variables analizadas fueron: edad, sexo, subtipo de leucemia y expresión de marcadores inmunológicos, cuyas asociaciones fueron analizadas con los estadígrafos Chi-cuadrado y coeficiente de correlación de Spearman. Resultados: se estudiaron 58 pacientes, 28 del sexo femenino y 30 del masculino. El grupo de edad predominante fue de 0 a 9 años con una mediana de 26 años. El subtipo M4 resultó el más frecuente (30,4 por ciento). Los subtipos M4 y M7 predominaron en niños, mientras que el M0, predominó en adultos, con diferencias estadísticamente significativas (p d¼0,05). La combinación de los antígenos panmieloides CD13 y CD33 se presentó en el 91 por ciento de los enfermos. Las combinaciones de CD13/CD33, CD14/CD15, CD33/CD14 y CD33/CD15 mostraron correlación significativa. En el 20,6 por ciento de los pacientes evaluados, fueron detectados, además, antígenos linfoides. No se encontraron diferencias significativas en cuanto al sexo y la edad. El antígeno CD7 fue el más expresado, seguido de los antígenos: CD3, CD20, CD22 y CD79, en igual proporción. Conclusiones: el inmunofenotipaje celular demostró ser un procedimiento útil para confirmar el diagnóstico morfológico y clínico de la leucemia mieloide aguda(AU)

Introduction: Acute myeloid leukemia (AML) is a heterogeneous group of diseases characterized by clonal expansion of myeloblasts; its classification involves several criteria, including the immunological one. Objective: To characterize the immunophenotype of patients with acute myeloid leukemia who were evaluated at the Institute of Hematology and Immunology. Methods: A descriptive study of patients diagnosed with this kind of leukemia, whose blood samples were processed at the Department of Immunology during 2008-2012. An immunocytochemical ultramicromethod that uses a panel of monoclonal antibodies specific for myeloid and lymphoid antigens was applied. The variables analyzed were age, sex, subtype of leukemia and expression of immunological markers; their association was analyzed with the Chi-square test and Spearman's rank correlation coefficient. Results: The study covered 58 patients; 28 were males and 30 females. The predominant age group was 0 - 9 years with a median of 26 years. M4 subtype was the most common (30,4 percent). M4 and M7 subtypes predominated in children, while M0 predominated in adults with statistically significant differences (p d¼ 0,05). The combination of pan-myeloid antigens CD13 and CD33 was present in 91 percent of patients. Combinations of CD13/CD33, CD14/CD15, CD33/CD14 and CD33/CD15 showed significant correlation. In 20,6 percent of patients tested, lymphoid antigens were also detected. In this group, no significant differences by gender and age. CD7 antigen was the most expressed followed by antigens: CD3, CD20, CD22 and CD79, in equal proportion. Conclusions: Immunophenotyping of leukemia cells appeared as a useful tool to confirm the morphological and clinical diagnosis of acute myeloid leukemia(AU)

Caracterización inmunofenotípica de pacientes con leucemia mieloide aguda / Immunophenotypic characterization of patients with acute myeloid leukemia

Introducción: la leucemia mieloide aguda incluye un grupo heterogéneo de neoplasias caracterizadas por una expansión clonal de mieloblastos, cuya clasificación involucra varios criterios, incluidos los inmunológicos. Objetivo: caracterizar el inmunofenotipo de los pacientes con leucemia mieloide aguda evaluados en el Instituto de Hematología e Inmunología. Métodos: se realizó un estudio descriptivo transversal de los pacientes diagnosticados con este tipo de leucemia, cuyas muestras de sangre fueron procesadas en el Departamento de Inmunología en el período 2008-2012. Se usó un ultramicrométodo inmunocitoquímico que utiliza un panel de anticuerpos monoclonales específicos de antígenos mieloides y linfoides. Las variables analizadas fueron: edad, sexo, subtipo de leucemia y expresión de marcadores inmunológicos, cuyas asociaciones fueron analizadas con los estadígrafos Chi-cuadrado y coeficiente de correlación de Spearman. Resultados: se estudiaron 58 pacientes, 28 del sexo femenino y 30 del masculino. El grupo de edad predominante fue de 0 a 9 años con una mediana de 26 años. El subtipo M4 resultó el más frecuente (30,4 por ciento). Los subtipos M4 y M7 predominaron en niños, mientras que el M0, predominó en adultos, con diferencias estadísticamente significativas (p d»0,05). La combinación de los antígenos panmieloides CD13 y CD33 se presentó en el 91 por ciento de los enfermos. Las combinaciones de CD13/CD33, CD14/CD15, CD33/CD14 y CD33/CD15 mostraron correlación significativa. En el 20,6 por ciento de los pacientes evaluados, fueron detectados, además, antígenos linfoides. No se encontraron diferencias significativas en cuanto al sexo y la edad. El antígeno CD7 fue el más expresado, seguido de los antígenos: CD3, CD20, CD22 y CD79, en igual proporción. Conclusiones: el inmunofenotipaje celular demostró ser un procedimiento útil para confirmar el diagnóstico morfológico y clínico de la leucemia mieloide aguda

Introduction: Acute myeloid leukemia (AML) is a heterogeneous group of diseases characterized by clonal expansion of myeloblasts; its classification involves several criteria, including the immunological one. Objective: To characterize the immunophenotype of patients with acute myeloid leukemia who were evaluated at the Institute of Hematology and Immunology. Methods: A descriptive study of patients diagnosed with this kind of leukemia, whose blood samples were processed at the Department of Immunology during 2008-2012. An immunocytochemical ultramicromethod that uses a panel of monoclonal antibodies specific for myeloid and lymphoid antigens was applied. The variables analyzed were age, sex, subtype of leukemia and expression of immunological markers; their association was analyzed with the Chi-square test and Spearman's rank correlation coefficient. Results: The study covered 58 patients; 28 were males and 30 females. The predominant age group was 0 - 9 years with a median of 26 years. M4 subtype was the most common (30,4 percent). M4 and M7 subtypes predominated in children, while M0 predominated in adults with statistically significant differences (p d» 0,05). The combination of pan-myeloid antigens CD13 and CD33 was present in 91 percent of patients. Combinations of CD13/CD33, CD14/CD15, CD33/CD14 and CD33/CD15 showed significant correlation. In 20,6 percent of patients tested, lymphoid antigens were also detected. In this group, no significant differences by gender and age. CD7 antigen was the most expressed followed by antigens: CD3, CD20, CD22 and CD79, in equal proportion. Conclusions: Immunophenotyping of leukemia cells appeared as a useful tool to confirm the morphological and clinical diagnosis of acute myeloid leukemia

Caracterización fenotípica de células madre mesenquimales humanas de médula ósea y tejido adiposo. Resultados preliminares / Phenotypic characterization of mesenchymal human stem cell from bone marrow and adipose tissue. preliminary results

Introducción:las células madre mesenquimales (CMM) poseen características fenotípicas y funcionales que les confieren un amplio potencial terapéutico por su posible uso en la terapia celular regenerativa, en el rechazo del trasplante alogénico y en enfermedades inflamatorias crónicas. Objetivo: evaluar la expresión de moléculas de membrana que permiten identificar la expresión de patrones moleculares característicos de CMM humanas mantenidas en cultivo. Métodos: se estudió la expresión fenotípica de células mononucleares procedentes de médula ósea obtenidas mediante aspiración medular, separadas por gradiente de Ficoll y cultivadas ex vivo entre los pases o subcultivos 3 y 16; y adipocitos cultivados procedentes de la extracción enzimática de tejido adiposo de donantes sanos. Se realizó doble marcaje para las moléculas CD34/CD45, CD34/CD90, CD34/CD117, y CD34/CD44. Resultados: en los resultados preliminares obtenidos se observó que las células cultivadas procedentes de médula ósea, entre los pases 4 y 8 de cultivo expresaron 45,13 por ciento de células CD34-/CD45- (doblemente negativas), lo que correspondió con el 25,24 por ciento de células CD34-/CD90+ y el 96,90 por ciento de CD34-/CD117-. En las células procedentes de cultivo de adipocitos se observó el 52,3 por ciento de CD34-/CD45- (doblemente negativas),12,31 por ciento de CD34-/CD90+,43,31 por ciento de CD34-/CD117- y 64,68 por ciento de CD34-/CD44+.Estos resultados sugieren que ambos cultivos se diferenciaron a CMM. Las CMM procedentes de adipocitos mostraron el 64,68 por ciento de células con expresión de la molécula de adhesión CD44 a la que se atribuyen propiedades funcionalescomo el asentamiento tisular. Conclusiones: estos resultados preliminares permiten corroborar que ambos métodos experimentales de cultivo son efectivos para la obtención de CMM con fines terapéuticos

Introduction:mesenchymal stem cells (MSCs) have phenotypic and functional characteristics whichgives them a broad therapeutic potential for possible use in regenerative cell therapy, allogeneic transplant rejection and chronic inflammatory diseases. Objective: to evaluate the expression of moleculemembranes expression to identify molecular patterns characteristic of human MSCs maintained in culture. Methods:the phenotypic expression of mononuclear cells from bone marrow wereobtained by bone marrow aspiration, separated by Ficoll and cultured ex vivo between passages or subcultures 3 and 16 and adipocytes cultured obtained from enzyme extraction of adipose tissue of healthy donor. Double staining was performed for molecules CD34/CD45, CD34/CD90, CD34/CD117 and CD34/CD44. Results:preliminary results showed that cultured mononuclear cells from bone marrow between passage 4 and 8 of culture expressed 45,13 percent CD34-/CD45- cells (double-negative), corresponding to 25,24 percent CD34-/CD90+ cells and 96,90 percent of CD34-/CD117-. Adipocytes from culture cells showed 52,3 percent CD34-/CD45- (double-negative), 12,31 percent cells CD34-/CD90+, 43,31 percent CD34-/CD117- (double-negative). Our results suggest that both cultures were differentiated to MSCs. Adipocytes from MSCs showed 64,68 percent of cells with expression of CD44 adhesion molecule conferring functional homing properties Conclusions:these preliminary results corroborate that the experimental methods used in cultivation are effective for obtaining MSCs with therapeutic purposes

Obtención y procesamiento de progenitores hematopoyéticos de sangre periférica para terapia celular en enfermedades angiológicas / Obtaining and processing of peripheral blood hematopoietic progenitor cells for cell therapy of angiological diseases

Se evaluó la seguridad y efectividad de un método manual de recolección de células progenitoras hematopoyéticas de sangre periférica movilizadas con factores estimuladores de colonias granulocíticas (FEC-G) de producción nacional (Leukocim y Hebervital). Se estudiaron 250 pacientes seleccionados para terapia celular en el Servicio de Angiología del Hospital General Docente Enrique Cabrera. La obtención y separación de las células mononucleares autólogas de sangre periférica (CMN-SP) se realizó mediante el método diseñado en el Instituto de Hematología e Inmunología. Para valorar la eficacia del método se analizaron en el concentrado obtenido las variables: contenido de células nucleadas, de células mononucleadas y de células CD 34+. Además, se determinó la viabilidad celular y la contaminación microbiológica. Se comprobó la eficiencia y seguridad del método de recolección y procesamiento para la obtención de un concentrado con un contenido de células mononucleares adecuado, sin complicaciones de importancia clínica. Se demostró la eficacia de los factores estimuladores de colonias granulocíticas empleados. Los efectos adversos de la movilización resultaron ligeros e independientes del factor estimulador utilizado(AU)

The safety and effectiveness of a manual collection method of peripheral blood hematopoietic progenitor cells mobilized by the Cuban-made granulocytic colony-stimulating factors (Leukocim and Hebervital) were evaluated. Two hundred patients, who had been selected for the cell therapy at the Angiology Service of Enrique Cabrera General Teaching Hospital, were studied. The method designed by the Institute of Hematology and Immunology served to obtain and separate autologous mononuclear cells from the peripheral blood. For the purpose of assessing the efficacy of this method, the variables contents of nucleate cells, of mononucleate cells and of CD 34+ cells were analyzed in the final concentrate. Additionally, the cell viability and the microbiological pollution were determined. The efficiency and safety of the collecting and processing method for obtaining one concentrate with adequate content of mononuclear cells and no significant clinical complications was confirmed. The efficacy of the Cuban granulocytic colony-stimulating factors was proved. The adverse effects of the mobilization were mild and unrelated to the used stimulating factor(AU)

Obtención y procesamiento de progenitores hematopoyéticos de sangre periférica para terapia celular en enfermedades angiológicas / Obtaining and processing of peripheral blood hematopoietic progenitor cells for cell therapy of angiological diseases

Se evaluó la seguridad y efectividad de un método manual de recolección de células progenitoras hematopoyéticas de sangre periférica movilizadas con factores estimuladores de colonias granulocíticas (FEC-G) de producción nacional (Leukocim y Hebervital). Se estudiaron 250 pacientes seleccionados para terapia celular en el Servicio de Angiología del Hospital General Docente Enrique Cabrera. La obtención y separación de las células mononucleares autólogas de sangre periférica (CMN-SP) se realizó mediante el método diseñado en el Instituto de Hematología e Inmunología. Para valorar la eficacia del método se analizaron en el concentrado obtenido las variables: contenido de células nucleadas, de células mononucleadas y de células CD 34+. Además, se determinó la viabilidad celular y la contaminación microbiológica. Se comprobó la eficiencia y seguridad del método de recolección y procesamiento para la obtención de un concentrado con un contenido de células mononucleares adecuado, sin complicaciones de importancia clínica. Se demostró la eficacia de los factores estimuladores de colonias granulocíticas empleados. Los efectos adversos de la movilización resultaron ligeros e independientes del factor estimulador utilizado

The safety and effectiveness of a manual collection method of peripheral blood hematopoietic progenitor cells mobilized by the Cuban-made granulocytic colony-stimulating factors (Leukocim and Hebervital) were evaluated. Two hundred patients, who had been selected for the cell therapy at the Angiology Service of Enrique Cabrera General Teaching Hospital, were studied. The method designed by the Institute of Hematology and Immunology served to obtain and separate autologous mononuclear cells from the peripheral blood. For the purpose of assessing the efficacy of this method, the variables contents of nucleate cells, of mononucleate cells and of CD 34+ cells were analyzed in the final concentrate. Additionally, the cell viability and the microbiological pollution were determined. The efficiency and safety of the collecting and processing method for obtaining one concentrate with adequate content of mononuclear cells and no significant clinical complications was confirmed. The efficacy of the Cuban granulocytic colony-stimulating factors was proved. The adverse effects of the mobilization were mild and unrelated to the used stimulating factor

Experiencia cubana con el uso terapéutico de células madre adultas / Cuban experience with the therapeutic use of adult stem cells

Las investigaciones básicas y clínicas realizadas en los últimos años sobre las células madre y sus posibilidades terapéuticas, son en la actualidad uno de los temas más excitantes de la medicina contemporánea. Ya se han obtenido importantes avances en el estudio y aplicación de las células madre adultas que muestran notables ventajas sobre las embrionarias, pues su manipulación resulta más simple, económica y se pueden obtener del propio individuo que va a ser tratado. Para la introducción en Cuba de la terapia celular regenerativa, en el Instituto de Hematología e Inmunología se seleccionaron como fuentes celulares las células madre adultas derivadas de la médula ósea y las movilizadas a la sangre periférica. Para facilitar la extensión del tratamiento a otros centros hospitalarios, se estandarizó una técnica para la movilización de las células madre hematopoyéticas a la sangre periférica, mediante un factor estimulador de colonias de granulocitos (Filgrastim, de producción nacional) y se desarrolló un método simple, económico y también más tolerable para los enfermos. De esta forma, se ha extendido la terapia celular a 6 provincias cubanas y hasta abril del año 2009 se habían tratado 563 casos con trasplante de células madre adultas autólogas, de los cuales el 81,7 por ciento corresponde a pacientes con enfermedades angiológicas, en los que se ha logrado disminuir significativamente la indicación de amputaciones mayores. También los resultados han sido muy prometedores en las lesiones óseas y procesos periodontales, entre otras enfermedades tratadas. Los resultados obtenidos hasta el momento se pueden considerar como un nuevo logro de la ciencia revolucionaria y de nuestros sistemas nacionales de salud y de ciencia y técnica. El método empleado es un proceder económico y factible para instituciones con recursos limitados(AU)

The basic and clinical researches carried out during past years on the stem cells and its therapeutic possibilities are at present times, one of the most interesting subjects of the contemporaneous medicine. There are advances in the study and application of adult stem cells showing remarkable advantages on the embryonic ones, since its handling is more simple, economic and they are obtained from the own subject to be treated. For the introduction in Cuba of the regenerative cellular therapy in the Institute of Hematology and Immunology the cellular sources selected were the adult stem cells derived from bone marrow and the mobilized ones to the peripheral blood. To make easy the expansion of treatment to other hospital centers, authors standardized a technique for the mobilization of the hematopoietic stem cells to peripheral blood using a granulocyte colony-stimulating factor (Filgrastim, of national production) developing a simple, economic and more tolerable method for patients. In this way, the cellular therapy has been expanded to 6 Cuban provinces and until April, 2009 562 cases with autologous adult stem cells transplant have been treated, from which the 81.7 percent to correspond to patients presenting with Angiology diseases with a significant reduction of major amputations. Also, the results have been very promising in the bone lesions and periodontal processes among other diseases treated. The results obtained until now may be considered as a new achievement of revolutionary science and of our national health systems and of science and technique. The method used is an economic and feasible procedure for the institutions with scarce resources(AU)

Experiencia cubana con el uso terapéutico de células madre adultas / Cuban experience with the therapeutic use of adult stem cells

Las investigaciones básicas y clínicas realizadas en los últimos años sobre las células madre y sus posibilidades terapéuticas, son en la actualidad uno de los temas más excitantes de la medicina contemporánea. Ya se han obtenido importantes avances en el estudio y aplicación de las células madre adultas que muestran notables ventajas sobre las embrionarias, pues su manipulación resulta más simple, económica y se pueden obtener del propio individuo que va a ser tratado. Para la introducción en Cuba de la terapia celular regenerativa, en el Instituto de Hematología e Inmunología se seleccionaron como fuentes celulares las células madre adultas derivadas de la médula ósea y las movilizadas a la sangre periférica. Para facilitar la extensión del tratamiento a otros centros hospitalarios, se estandarizó una técnica para la movilización de las células madre hematopoyéticas a la sangre periférica, mediante un factor estimulador de colonias de granulocitos (Filgrastim, de producción nacional) y se desarrolló un método simple, económico y también más tolerable para los enfermos. De esta forma, se ha extendido la terapia celular a 6 provincias cubanas y hasta abril del año 2009 se habían tratado 563 casos con trasplante de células madre adultas autólogas, de los cuales el 81,7 por ciento corresponde a pacientes con enfermedades angiológicas, en los que se ha logrado disminuir significativamente la indicación de amputaciones mayores. También los resultados han sido muy prometedores en las lesiones óseas y procesos periodontales, entre otras enfermedades tratadas. Los resultados obtenidos hasta el momento se pueden considerar como un nuevo logro de la ciencia revolucionaria y de nuestros sistemas nacionales de salud y de ciencia y técnica. El método empleado es un proceder económico y factible para instituciones con recursos limitados

The basic and clinical researches carried out during past years on the stem cells and its therapeutic possibilities are at present times, one of the most interesting subjects of the contemporaneous medicine. There are advances in the study and application of adult stem cells showing remarkable advantages on the embryonic ones, since its handling is more simple, economic and they are obtained from the own subject to be treated. For the introduction in Cuba of the regenerative cellular therapy in the Institute of Hematology and Immunology the cellular sources selected were the adult stem cells derived from bone marrow and the mobilized ones to the peripheral blood. To make easy the expansion of treatment to other hospital centers, authors standardized a technique for the mobilization of the hematopoietic stem cells to peripheral blood using a granulocyte colony-stimulating factor (Filgrastim, of national production) developing a simple, economic and more tolerable method for patients. In this way, the cellular therapy has been expanded to 6 Cuban provinces and until April, 2009 562 cases with autologous adult stem cells transplant have been treated, from which the 81.7 percent to correspond to patients presenting with Angiology diseases with a significant reduction of major amputations. Also, the results have been very promising in the bone lesions and periodontal processes among other diseases treated. The results obtained until now may be considered as a new achievement of revolutionary science and of our national health systems and of science and technique. The method used is an economic and feasible procedure for the institutions with scarce resources

Aislamiento de células mononucleares de sangre periférica para trasplante de células madre: método simplificado / Isolation of mononuclear cells of peripheral blood for stem cell transplant: simplified method

En los últimos años el tema de las células madre ha despertado creciente interés por su potencial terapéutico en enfermedades que hasta el momento no tienen un tratamiento efectivo. Se realizó un estudio prospectivo y exploratorio en pacientes con arteriosclerosis obliterante de miembros inferiores, en el que se evaluó la seguridad y efectividad de un método manual de recolección y procesamiento de células mononucleares y de células CD34+ a partir de sangre periférica movilizada. La sangre se procesó en sistemas cerrados de recolección, utilizando el hidroxietilalmidón como potenciador de la sedimentación eritrocitaria. Los pacientes fueron tratados con factor estimulador de colonias granulocítico en dosis total de 40ìcg/kg de peso durante 2 días, y después si el conteo de leucocitos era superior a 20 x 109/L se procedió a la autodonación. Para valorar la eficacia del método se analizaron las cantidades de células nucleadas, de células mononucleares y de células CD 34+ en el concentrado celular; se determinó la viabilidad celular y además se hizo el estudio microbiológico del material obtenido. Se demostró que el método es eficaz y seguro, ya que logra niveles celulares adecuados, con elevada viabilidad y ausencia de contaminación bacteriana. Por otra parte, es sencillo y de bajo costo, lo que permite su extensión a otros centros de salud, en particular a los de menos recursos. Esto facilita que un mayor número de pacientes se puedan beneficiar con el tratamiento a base de células madre(AU)

In the last years, the topic of stem cells has arisen an increasing interest for its therapeutic potential in diseases that have not an effective treatment so far. A prospsective and exploratory study was conducted in patients with obliterant atherosclerosis of the lower limbs to evaluate the safety and effectiveness of a manual method of collection and processing of mononuclear cells and of CD34+ cells, starting from mobilized peripheral blood. The blood was processed in closed collection systems, using hydroxyethyl starch as a potentiator of erythrocyte sedimentation. The patients were treated with granulocyte colony stimulating factor at total doses of 40µcg/kg of weight during 2 days. Self-donation was performed when the leukocyte count was higher than 20 x 109/L. To assess the efficacy of the method, the amounts of nucleated cells, of mononuclear cells and of CD 34+ cells in the cellular concentrate were analyzed. Cellular viability was determined and a microbiological study of the material obtained was conducted. It was proved that the method is efficient and safe, since adequate cellular levels with a high viability and absence of bacterial contamination are attained. On the other hand, it is simple and cheap, which allows its application in other health centres, particularly in those with less resources. This makes possible that more patients benefit from the stem cell treatment(AU)

Aislamiento de células mononucleares de sangre periférica para trasplante de células madre: método simplificado / Isolation of mononuclear cells of peripheral blood for stem cell transplant: simplified method

En los últimos años el tema de las células madre ha despertado creciente interés por su potencial terapéutico en enfermedades que hasta el momento no tienen un tratamiento efectivo. Se realizó un estudio prospectivo y exploratorio en pacientes con arteriosclerosis obliterante de miembros inferiores, en el que se evaluó la seguridad y efectividad de un método manual de recolección y procesamiento de células mononucleares y de células CD34+ a partir de sangre periférica movilizada. La sangre se procesó en sistemas cerrados de recolección, utilizando el hidroxietilalmidón como potenciador de la sedimentación eritrocitaria. Los pacientes fueron tratados con factor estimulador de colonias granulocítico en dosis total de 40ìcg/kg de peso durante 2 días, y después si el conteo de leucocitos era superior a 20 x 109/L se procedió a la autodonación. Para valorar la eficacia del método se analizaron las cantidades de células nucleadas, de células mononucleares y de células CD 34+ en el concentrado celular; se determinó la viabilidad celular y además se hizo el estudio microbiológico del material obtenido. Se demostró que el método es eficaz y seguro, ya que logra niveles celulares adecuados, con elevada viabilidad y ausencia de contaminación bacteriana. Por otra parte, es sencillo y de bajo costo, lo que permite su extensión a otros centros de salud, en particular a los de menos recursos. Esto facilita que un mayor número de pacientes se puedan beneficiar con el tratamiento a base de células madre.

In the last years, the topic of stem cells has arisen an increasing interest for its therapeutic potential in diseases that have not an effective treatment so far. A prospsective and exploratory study was conducted in patients with obliterant atherosclerosis of the lower limbs to evaluate the safety and effectiveness of a manual method of collection and processing of mononuclear cells and of CD34+ cells, starting from mobilized peripheral blood. The blood was processed in closed collection systems, using hydroxyethyl starch as a potentiator of erythrocyte sedimentation. The patients were treated with granulocyte colony stimulating factor at total doses of 40µcg/kg of weight during 2 days. Self-donation was performed when the leukocyte count was higher than 20 x 109/L. To assess the efficacy of the method, the amounts of nucleated cells, of mononuclear cells and of CD 34+ cells in the cellular concentrate were analyzed. Cellular viability was determined and a microbiological study of the material obtained was conducted. It was proved that the method is efficient and safe, since adequate cellular levels with a high viability and absence of bacterial contamination are attained. On the other hand, it is simple and cheap, which allows its application in other health centres, particularly in those with less resources. This makes possible that more patients benefit from the stem cell treatment.