MIR4435-2HG as a possible novel predictive biomarker of chemotherapy response and death in pediatric B-cell ALL.

Introduction: Although B-cell acute lymphoblastic leukemia (B-cell ALL) survival rates have improved in recent years, Hispanic children continue to have poorer survival rates. There are few tools available to identify at the time of diagnosis whether the patient will respond to induction therapy. Our goal was to identify predictive biomarkers of treatment response, which could also serve as prognostic biomarkers of death, by identifying methylated and differentially expressed genes between patients with positive minimal residual disease (MRD+) and negative minimal residual disease (MRD-). Methods: DNA and RNA were extracted from tumor blasts separated by immunomagnetic columns. Illumina MethlationEPIC and mRNA sequencing assays were performed on 13 bone marrows from Hispanic children with B-cell ALL. Partek Flow was used for transcript mapping and quantification, followed by differential expression analysis using DEseq2. DNA methylation analyses were performed with Partek Genomic Suite and Genome Studio. Gene expression and differential methylation were compared between patients with MRD-/- and MRD+/+ at the end of induction chemotherapy. Overexpressed and hypomethylated genes were selected and validated by RT-qPCR in samples of an independent validation cohort. The predictive ability of the genes was assessed by logistic regression. Survival and Cox regression analyses were performed to determine the association of genes with death. Results: DAPK1, BOC, CNKSR3, MIR4435-2HG, CTHRC1, NPDC1, SLC45A3, ITGA6, and ASCL2 were overexpressed and hypomethylated in MRD+/+ patients. Overexpression was also validated by RT-qPCR. DAPK1, BOC, ASCL2, and CNKSR3 can predict refractoriness, but MIR4435-2HG is the best predictor. Additionally, higher expression of MIR4435-2HG increases the probability of non-response, death, and the risk of death. Finally, MIR4435-2HG overexpression, together with MRD+, are associated with poorer survival, and together with overexpression of DAPK1 and ASCL2, it could improve the risk classification of patients with normal karyotype. Conclusion: MIR4435-2HG is a potential predictive biomarker of treatment response and death in children with B-cell ALL.

Untargeted metabolomic and lipidomic analyses reveal lipid dysregulation in the plasma of acute leukemia patients.

Acute leukemias (AL) are aggressive neoplasms with high mortality rates. Metabolomics and oxidative status have emerged as important tools to identify new biomarkers with clinical utility. To identify the metabolic differences between healthy individuals (HI) and patients with AL, a multiplatform untargeted metabolomic and lipidomic approach was conducted using liquid and gas chromatography coupled with quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS or GC-QTOF-MS). Additionally, the total antioxidant capacity (TAC) was measured. A total of 20 peripheral blood plasma samples were obtained from patients with AL and 18 samples from HI. Our analysis revealed 135 differentially altered metabolites in the patients belonging to 12 chemical classes; likewise, the metabolic pathways of glycerolipids and sphingolipids were the most affected in the patients. A decrease in the TAC of the patients with respect to the HI was evident. This study conducted with a cohort of Colombian patients is consistent with observations from other research studies that suggest dysregulation of lipid compounds. Furthermore, metabolic differences between patients and HI appear to be independent of lifestyle, race, or geographic location, providing valuable information for future advancements in understanding the disease and developing more global therapies.

IL-8 in bone marrow and peripheral blood of patients with B-cell acute lymphoblastic leukemia is associated with high regulatory T cell counts, degree of tumor infiltration and expression of CXCR1 in blasts.

INTRODUCTION: Regulatory T cells (Treg cells) in a tumor environment and the expression of forkhead box P3 (FOXP3) in tumor cells have been associated with a poor prognosis. There are few studies evaluating Treg cells and FOXP3 in B-cell acute lymphoblastic leukemia (B-cell ALL). This study aimed to evaluate the frequencies of Treg cells in bone marrow (BM) and peripheral blood (PB) of patients with B-cell ALL and to determine their associations with the circulating cytokine profile and the expression of CXCR1 (IL-8 receptor) in Treg cells, as well as to compare FOXP3 expression in blasts of patients with B-cell ALL and normal lymphoid precursors. METHODS: Samples of BM and PB from patients with B-cell ALL and healthy controls were studied. Treg cells, cytokines, FOXP3 and CXCR1 were evaluated using flow cytometry and analyzed. RESULTS: A total of 20 patients with B-cell ALL and 10 healthy controls were included. In B-cell ALL patients, Treg cell frequencies increased significantly, with higher percentages in the PB. Absolute Treg cell counts were associated with absolute blast counts in the BM and PB and with an IL-8 concentration. The IL-8 and IL-6 levels were associated with the CXCR1 expression in PB Treg cells. In addition, a greater expression of FOXP3 was observed in leukemic blasts than in normal lymphoid precursors. CONCLUSIONS: These results suggest that the presence of Treg cells and cytokines in the tumor environment may correspond to mechanisms to evade the immune response. For that reason, it would be important to monitor these parameters in B-cell ALL to establish their effect on the disease prognosis.

Plant-derived extracts and metabolic modulation in leukemia: a promising approach to overcome treatment resistance.

Leukemic cells acquire complex and often multifactorial mechanisms of resistance to treatment, including various metabolic alterations. Although the use of metabolic modulators has been proposed for several decades, their use in clinical practice has not been established. Natural products, the so-called botanical drugs, are capable of regulating tumor metabolism, particularly in hematopoietic tumors, which could partly explain the biological activity attributed to them for a long time. This review addresses the most recent findings relating to metabolic reprogramming-Mainly in the glycolytic pathway and mitochondrial activity-Of leukemic cells and its role in the generation of resistance to conventional treatments, the modulation of the tumor microenvironment, and the evasion of immune response. In turn, it describes how the modulation of metabolism by plant-derived extracts can counteract resistance to chemotherapy in this tumor model and contribute to the activation of the antitumor immune system.

In Vitro Evaluation of Anti-Rotaviral Activity and Intestinal Toxicity of a Phytotherapeutic Prototype of Achyrocline bogotensis (Kunth) DC.

Viruses represent the primary etiologic agents (70-80%) of acute diarrheal disease (ADD), and rotavirus (RV) is the most relevant one. Currently, four rotavirus vaccines are available. However, these vaccines do not protect against emerging viral strains or are not available in low-income countries. To date, there are no approved drugs available against rotavirus infection. In this study, we evaluated the in vitro anti-rotaviral activity and intestinal toxicity of a phytotherapeutic prototype obtained from Achyrocline bogotensis (Kunth) DC. (PPAb); medicinal plant that contains compounds that inhibit the rotavirus replication cycle. Virucidal and viral yield reduction effects exerted by the PPAb were evaluated by immunocytochemistry and flow cytometry. Furthermore, the toxic impact of the PPAb was evaluated in polarized human intestinal epithelial C2BBe1 cells in terms of cytotoxicity, loss of cytoplasmic membrane asymmetry, and DNA fragmentation by MTT and fluorometry. PPAb concentrations under 0.49 mg/mL exerted significant virucidal and viral yield reduction activities, and concentrations under 16 mg/mL neither reduced cell viability, produced DNA fragmentation, nor compromised the C2BBe1cell membrane stability after 24-h incubation. Based on these results, the evaluated phytotherapeutic prototype of Achyrocline bogotensis might be considered as a promising alternative to treat ADD caused by rotavirus.

Randomized double-blind clinical study in patients with COVID-19 to evaluate the safety and efficacy of a phytomedicine (P2Et).

Background: It has been proposed that polyphenols can be used in the development of new therapies against COVID-19, given their ability to interfere with the adsorption and entrance processes of the virus, thus disrupting viral replication. Seeds from Caesalpinia spinosa, have been traditionally used for the treatment of inflammatory pathologies and respiratory diseases. Our team has obtained an extract called P2Et, rich in polyphenols derived from gallic acid with significant antioxidant activity, and the ability to induce complete autophagy in tumor cells and reduce the systemic inflammatory response in animal models. Methods: In this work, a phase II multicenter randomized double-blind clinical trial on COVID-19 patients was designed to evaluate the impact of the P2Et treatment on the clinical outcome and the immunological parameters related to the evolution of the disease. The Trial was registered with the number No. NCT04410510*. A complementary study in an animal model of lung fibrosis was carried out to evaluate in situ lung changes after P2Et in vivo administration. The ability of P2Et to inhibit the viral load of murine and human coronaviruses in cellular models was also evaluated. Results: Patients treated with P2Et were discharged on average after 7.4 days of admission vs. 9.6 days in the placebo group. Although a decrease in proinflammatory cytokines such as G-CSF, IL-15, IL-12, IL-6, IP10, MCP-1, MCP-2 and IL-18 was observed in both groups, P2Et decreased to a greater extent G-CSF, IL-6 and IL-18 among others, which are related to lower recovery of patients in the long term. The frequency of T lymphocytes (LT) CD3+, LT double negative (CD3+CD4-CD8-), NK cells increased in the P2Et group where the population of eosinophils was also significantly reduced. In the murine bleomycin model, P2Et also reduced lung inflammation and fibrosis. P2Et was able to reduce the viral replication of murine and human coronaviruses in vitro, showing its dual antiviral and anti-inflammatory role, key in disease control. Conclusions: Taken together these results suggest that P2Et could be consider as a good co-adjuvant in the treatment of COVID-19. Clinical trail registration: https://clinicaltrials.gov/ct2/show/NCT04410510, identifier: NCT04410510.

Assessment of Placental Extracellular Vesicles-Associated Fas Ligand and TNF-Related Apoptosis-Inducing Ligand in Pregnancies Complicated by Early and Late Onset Preeclampsia.

Preeclampsia (PE) is a hypertensive disorder that affects 2-8% of pregnancies and is one of the main causes of fetal, neonatal, and maternal mortality and morbidity worldwide. Although PE etiology and pathophysiology remain unknown, there is evidence that the hyperactivation of maternal immunity cells against placental cells triggers trophoblast cell apoptosis and death. It has also been reported that placenta-derived extracellular vesicles (EV) carry Fas ligand (FasL) and Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trigger apoptosis in Jurkat T cells. This study aimed to quantify and compare FasL and TRAIL expression in EV derived from cultures of placenta explants from women with PE (early versus late) and women with uncomplicated pregnancies. Also, the study assessed EV capacity to induce apoptosis in Jurkat T cells. The authors isolated EV from placenta explant cultures, quantified FasL and TRAIL using ELISA, and analyzed EV apoptosis-inducing capability by flow cytometry. Results showed increased FasL and TRAIL in EV derived from placenta of women with PE, and increased EV apoptosis-inducing capability in Jurkat T cells. These results offer supporting evidence that EV FasL and TRAIL play a role in the pathophysiology of PE.

Facilitadores y barreras percibidos en la práctica de la actividad física en adolescentes escolarizados en Piedecuesta (Santander), en 2016: análisis cualitativo / Perceived facilitators and barriers to physical activity in adolescents in Piedecuesta (Santander), in 2016: qualitative analysis / Facilitadores e barreiras evidentes na prática da atividade física nos adolescentes em idade escolar em Piedecuesta Santander, em 2016: Análise qualitativa

Resumen Objetivo: Describir la percepción que tienen adolescentes escolarizados entre 12 y 16 años, en Piedecuesta (Santander), en 2016, sobre los facilitadores y las barreras que influyen en la práctica de la actividad física. Metodología: Análisis cualitativo desde un enfoque fenomenológico, con la participación de 20 adolescentes escolarizados que, mediante entrevistas semiestructuradas y un grupo focal, conversaron sobre la práctica, los beneficios, los entornos y la influencia y la compañía en relación con la actividad física, para identificar barreras y facilitadores. Resultados: La práctica de la actividad física se limitaba a las clases de educación física en la mayoría de los casos. El compañero principal en la práctica extracurricular era el padre del mismo sexo. La actividad física representaba una oportunidad para el disfrute, mejorar la salud física y mental, así como la contextura corporal, y para romper con la rutina diaria. La falta de tiempo por compromisos académicos fue la razón principal para no realizar actividad física. Los lugares públicos no siempre estaban disponibles, a menudo no se consideraban seguros. En la escuela, el espacio y las instalaciones eran inadecuados, y el uso estaba restringido. Conclusiones: La escuela es un punto de convergencia importante para la práctica de la actividad física, lo que sugiere la necesidad de fortalecer el plan de estudios al respecto; este entorno es un facilitador potencial para aumentar el nivel de actividad física en adolescentes. También se percibe como barrera, dado que la práctica interfiere con las actividades académicas. Las mejoras en la seguridad y el atractivo estético en el entorno comunitario se visibilizan como un facilitador para promover la actividad física.

Abstract Objective: Describe the perception of adolescents aged between 12 and 16 years, in Piedecuesta (Santander), in 2016, about the facilitators and barriers that influence physical activity. Methodology: Qualitative analysis from a phenomenological approach, with the participation of 20 adolescents who talked about the practice, benefits, environments, influence and company in connection with physical activity , to identify barriers and facilitators, through semi-structured interviews and a focus group. Results: Physical activity was limited to physical education classes in most cases. The main companion in extracurricular physical activity was the parent of the same sex. Physical activity represented an opportunity for enjoyment, improvement of physical and mental health, as well as body composition, and to have a break in their daily routine. Lack of time due to academic commitments was the main reason for not doing physical activity. Public places were not always available and they were often not considered to be safe. Space and facilities at school were inadequate, and their use was restricted.. Conclusions: The school is an important point of convergence for doing physical activity, suggesting the need to strengthen the curriculum in this respect; this environment is a potential facilitator to increasing the level of physical activity in adolescents. It can also be seen as a barrier, given that the practice interferes with academic activities. Improvements in safety and aesthetic appeal in the surrounding community are seen as a facilitator to promoting physical activity.

Resumo Objetivo: Descrever a percepção que os adolescentes entre 12 e 17 anos de Piedecuesta em Santander, tiveram em 2016 sobre os facilitadores e as barreiras que influenciaram na prática da atividade física. Metodologia: Análise qualitativa partindo de um foco fenomenológico, com a participação de 20 adolescentes em idade escolar que, através de entrevistas semiestruturadas e um grupo focal, conversaram sobre a prática, os benefícios, os entornos, a influência e o acompanhamento relacionados com a atividade física, para identificar barreiras e facilitadores. Resultados: Na maioria dos casos, a prática da atividade física limitava-se às aulas de educação física. O companheiro principal na prática extracurricular era o genitor ou genitora do mesmo sexo. A atividade física representava uma oportunidade para disfrutar, melhorar a saúde física e mental, além da estrutura corporal e para sair da rotina diária. A falta de tempo devido às responsabilidades escolares foi a principal razão para não realizarem atividade física. Os lugares públicos nem sempre estavam disponíveis e geralmente não são considerados seguros. Na escola, o espaço e as instalações eram inadequados e o uso estava restrito. Conclusões: A escola é um importante ponto de convergência para a prática da atividade física, o que determina a necessidade de fortalecer o currículo relacionado; esse ambiente é um facilitador potencial para aumentar o nível de atividade física nos adolescentes. Também se percebe como barreira, já que a prática interfere com as atividades escolares. As melhorias na segurança e um espaço comunitário mais atraente do ponto de vista estético são determinantes como facilitadores para a promoção da atividade física.

Influence of Dose Intensity in Consolidation with HIDAC and Other Clinical and Biological Parameters in the Survival of AML.

BACKGROUND: The impact of the dose intensity administered in consolidation in Latin America is unknown. This study aimed to evaluate the relative dose intensity (RDI) in consolidation and its impact in overall survival. METHODS: A retrospective study of 86 patients with AML who were diagnosed between 2010 and 2016 with a 2-year follow-up in a fourth-level Colombian hospital was carried out. Clinical characteristics were reported, Kaplan-Meier was used for estimating the overall survival, and Cox regression was used for multivariate analysis. RESULTS: The median overall survival (OS) was 20.83 months, and the median event-free survival (EFS) was 16.83 months. 64.3% of the patients achieved remission after the 7 + 3 chemotherapy induction treatment. Patients under 30 years of age, with white blood cell counts less than 100.000 cells/mm3 who responded to induction treatment had a better OS. Additionally, patients receiving an RDI greater than 0.75 of the planned consolidation dose had better survival. The prognostic variables with impact in the OS were the leukocyte count in peripheral blood at diagnosis, the RDI in consolidation treatment with HIDAC and the response obtained after induction. CONCLUSION: This retrospective study allowed us to know the epidemiology of AML in a reference Colombian Hospital. Additionally, in our knowledge, it is the first study that reports the RDI in consolidation with HIDAC in Latin America as a prognostic factor that directly impacts the OS.

Preferential Activity of Petiveria alliacea Extract on Primary Myeloid Leukemic Blast.

The need for new therapeutic approaches to improve the response in acute leukemia (AL), either by directing therapy or with new therapeutic alternatives, has been a research and clinical interest topic. We evaluated whether blasts from AL patients were sensitive ex vivo to the induction chemotherapy and whether the extracts of Petiveria alliacea (Anamu SC) and Caesalpinia spinosa (P2Et) modulated the sensitivity of leukemic cells to death. Bone marrow samples were taken from 26 patients with de novo AL and 6 in relapse, and the cytotoxicity of the extracts alone or in combination with the chemotherapeutic was evaluated by XTT. Patients were classified as good (GR) and bad responders (BR) according to the ex vivo test. 70.5% of the GR patients to the ex vivo test achieved postinduction remission to induction chemotherapy with a median overall survival of 12.50 months versus 7.23 months in the two groups. Furthermore, it was found that the ex vivo response to extracts and chemotherapeutics is heterogeneous and shows an exclusive pattern between the extracts, Anamu being the more effective in inducing cell death. The combination of extracts with chemotherapeutic agents showed synergistic or antagonistic effects in the patients' blasts. These results show that the ex vivo evaluation of the sensitivity to induction drugs using primary blasts from patients exhibits a correlation with the response to induction chemotherapy in patients. These analyses would allow establishing a system to predict response to treatment and determine ex vivo susceptibility to new therapies under development, among which is phytotherapeutics.

[Concordance of the assessment of pubertal development with the Tanner scale between adolescents and a trained physician]. / Concordancia en la evaluación del desarrollo puberal mediante la escala de Tanner entre adolescentes y un médico entrenado.

OBJECTIVES.: To assess the concordance between self-assessment of pubertal development with the Tanner scale performed by adolescents compared to the assessment performed by a trained physician. MATERIALS AND METHODS.: As part of the SIMAC project, data was collected on 244 adolescents. At baseline, we included an anthropometric and pubertal development assessment, with a self-assessment by the participants. RESULTS.: We included 229 teenagers from 12 to 17 years old. The agreement between the self-assessment and the clinical evaluation with the Tanner scale presented an 88.3% agreement and a weighted kappa coefficient of 0.554. The self-assessment in women showed a good agreement ( kappaweighted=0.653) and in men a moderate agreement (kappaweighted=0.464). The only variable with a significant influence on the agreement ability was gender; the disagreement probability in males was 63% greater than in females, regardless of age, height, or weight (95% CI: 1,18-2, 26). CONCLUSIONS: . Self-assessment was not accurate enough to replace medical examination, especially in males. More research is needed on this subject considering the impact that self-assessment of sexual development could have.

OBJETIVOS.: Evaluar la concordancia entre la autoevaluación del desarrollo puberal mediante la escala de Tanner realizada por adolescentes, comparado con la evaluación efectuada por un médico entrenado. MATERIALES Y MÉTODOS.: Los datos de 244 adolescentes fueron recolectados como parte del proyecto SIMAC. En la línea de base incluimos una evaluación antropométrica y del desarrollo puberal, con una autoevaluación por parte de los participantes. RESULTADOS.: Incluimos 229 adolescentes de 12 a 17 años. La concordancia entre la autoevaluación y la evaluación clínica de la escala de Tanner presentó un acuerdo obtenido de 88,3% y un coeficiente kappa ponderado de 0,554. La autoevaluación en mujeres demostró una concordancia buena (kappapond=0,653), y en varones una concordancia moderada (kappapond=0,464). La única variable con una influencia significativa sobre la capacidad de acuerdo fue el sexo; la probabilidad de desacuerdo en los varones fue 63% mayor que en las mujeres, independiente de edad, talla y peso (IC 95%:1,18-2,26). CONCLUSIONES.: La autoevaluación no fue lo suficientemente precisa para reemplazar el examen médico, sobre todo en los varones. Se requieren más estudios en relación con este tema, reconociendo el impacto que podría llegar a tener la autoevaluación del desarrollo sexual.

Concordancia en la evaluación del desarrollo puberal mediante la escala de Tanner entre adolescentes y un médico entrenado / Concordance of the assessment of pubertal development with the Tanner scale between adolescents and a trained physician

RESUMEN Objetivos. Evaluar la concordancia entre la autoevaluación del desarrollo puberal mediante la escala de Tanner realizada por adolescentes, comparado con la evaluación efectuada por un médico entrenado. Materiales y métodos. Los datos de 244 adolescentes fueron recolectados como parte del proyecto SIMAC. En la línea de base incluimos una evaluación antropométrica y del desarrollo puberal, con una autoevaluación por parte de los participantes. Resultados. Incluimos 229 adolescentes de 12 a 17 años. La concordancia entre la autoevaluación y la evaluación clínica de la escala de Tanner presentó un acuerdo obtenido de 88,3% y un coeficiente kappa ponderado de 0,554. La autoevaluación en mujeres demostró una concordancia buena (kappapond=0,653), y en varones una concordancia moderada (kappapond=0,464). La única variable con una influencia significativa sobre la capacidad de acuerdo fue el sexo; la probabilidad de desacuerdo en los varones fue 63% mayor que en las mujeres, independiente de edad, talla y peso (IC 95%:1,18-2,26). Conclusiones. La autoevaluación no fue lo suficientemente precisa para reemplazar el examen médico, sobre todo en los varones. Se requieren más estudios en relación con este tema, reconociendo el impacto que podría llegar a tener la autoevaluación del desarrollo sexual.

ABSTRACT Objectives. To assess the concordance between self-assessment of pubertal development with the Tanner scale performed by adolescents compared to the assessment performed by a trained physician. Materials and Methods. As part of the SIMAC project, data was collected on 244 adolescents. At baseline, we included an anthropometric and pubertal development assessment, with a self-assessment by the participants. Results. We included 229 teenagers from 12 to 17 years old. The agreement between the self-assessment and the clinical evaluation with the Tanner scale presented an 88.3% agreement and a weighted kappa coefficient of 0.554. The self-assessment in women showed a good agreement ( kappaweighted=0.653) and in men a moderate agreement (kappaweighted=0.464). The only variable with a significant influence on the agreement ability was gender; the disagreement probability in males was 63% greater than in females, regardless of age, height, or weight (95% CI: 1,18-2, 26). Conclusions . Self-assessment was not accurate enough to replace medical examination, especially in males. More research is needed on this subject considering the impact that self-assessment of sexual development could have.

May Measurement Month 2017: an analysis of blood pressure screening results in Colombia-Americas.

Despite the availability of efficient methods to evaluate blood pressure (BP) and of safe and efficient medication to treat and control hypertension, the levels of awareness, treatment and control are very low globally, particularly in low- and middle-income countries. To highlight the importance of improving these rates, the International Society of Hypertension (ISH) endorsed by the World Hypertension League have implemented the May Measurement Month initiative. We present here the results obtained in Colombia. The Fundación Oftalmológica de Santander (FOSCAL) led the implementation of this strategy in Colombia and 11 departments participated. The data collection followed the guidelines of the ISH. The information collected was compiled for the report generation and the submission to the Technical Secretariat of the ISH. Data cleaning was performed locally by FOSCAL. Data were collated and analysed centrally. A total of 22 258 participants (58.8% female) were included in the analysis. Mean age was 40.9 ± 17.7 years. Age and sex-standardized BP excluding participants receiving BP medications was 118/74.3 mmHg, and in those on treatment 125/78 mmHg. High BP was present in 5036 (22.8%) individuals, 1637 of 18 644 (8.8%) who were not receiving anti-hypertensive medications were hypertensive, and 961 of 3359 (28.6%) receiving treatment were not controlled. These results highlight the need to develop innovative promotion strategies at individual and population levels to increase the awareness of the importance of BP, and the consequences of not having well-controlled hypertension. This initiative is an effective and easy to implement strategy that should be maintained in the coming years.

Loss of T-Cell Multifunctionality and TCR-Vß Repertoire Against Epstein-Barr Virus Is Associated With Worse Prognosis and Clinical Parameters in HIV+ Patients.

Epstein-Barr virus (EBV) is an oncogenic virus associated with the development of aggressive and poor-prognosis B-cell lymphomas in patients infected with human immunodeficiency virus (HIV+ patients). The most important risk factors for these malignancies include immune dysfunction, chronic immune activation, and loss of T-cell receptor (TCR) repertoire. The combination of all these factors can favor the reactivation of EBV, malignant cell transformation, and clinical progression toward B-cell lymphomas. The overarching aim of this study was to evaluate the frequency, phenotype, functionality, and distribution of TCR clonotypes for EBV-specific T-cell subpopulations in HIV+ patients at different clinical stages and for HIV+ patients with B-cell lymphoma, as well as to establish their association with clinical variables of prognostic value. Factors were studied in 56 HIV+ patients at different clinical stages and in six HIV+ subjects with diagnosed B-cell lymphoma. We found a significant decrease in all subpopulations of EBV-specific CD4+ T cells from HIV+ patients at stage 3 and with B-cell lymphoma. EBV-specific effector CD8+ T cells, particularly effector memory cells, were also reduced in HIV+ patients with B-cell lymphoma. Interestingly, these cells were unable to produce IFN-γ and lacked multifunctionality in HIV+ patients. The TCR-Vß repertoire, which is key for protection against EBV in healthy individuals, was less diverse in HIV+ patients due to a lower frequency of TCR-Vß2+, Vß4+, Vß7.1+, Vß9+, Vß13.6+, Vß14+, Vß17+, Vß22+ CD4+, Vß14+, and Vß17+ CD8+ T cells. HIV+ patients with positive plasma EBV loads (EBV+HIV+) had a noteworthy decrease in the levels of both TNF-α+ and multifunctional TNF-α+/IL-2+ and TNF-α+/IFN-γ+ CD8+ T cells. Altogether, our findings demonstrate that HIV+ patients have significant alterations in the immune response to EBV (poor-quality immunity) that can favor viral reactivation, escalating the risk for developing EBV-associated B-cell lymphomas.

Obesity and Preeclampsia: Common Pathophysiological Mechanisms.

Preeclampsia is a disorder specific of the human being that appears after 20 weeks of pregnancy, characterized by new onset of hypertension and proteinuria. Abnormal placentation and reduced placental perfusion associated to impaired trophoblast invasion and alteration in the compliance of uterine spiral arteries are the early pathological findings that are present before the clinical manifestations of preeclampsia. Later on, the endothelial and vascular dysfunction responsible of the characteristic vasoconstriction of preeclampsia appear. Different nutritional risk factors such as a maternal deficit in the intake of calcium, protein, vitamins and essential fatty acids, have been shown to play a role in the genesis of preeclampsia, but also an excess of weight gain during pregnancy or a pre-pregnancy state of obesity and overweight, which are associated to hyperinsulinism, insulin resistance and maternal systemic inflammation, are proposed as one of the mechanism that conduce to endothelial dysfunction, hypertension, proteinuria, thrombotic responses, multi-organ damage, and high maternal mortality and morbidity. Moreover, it has been demonstrated that pregnant women that suffer preeclampsia will have an increased risk of future cardiovascular disease and related mortality in their later life. In this article we will discuss the results of studies performed in different populations that have shown an interrelationship between obesity and overweight with the presence of preeclampsia. Moreover, we will review some of the common mechanisms that explain this interrelationship, particularly the alterations in the L-arginine/nitric oxide pathway as a crucial mechanism that is common to obesity, preeclampsia and cardiovascular diseases.

Study of body fluid samples using flow cytometry: Six years of experience at the Hospital Universitario San Ignacio - Pontificia Universidad Javeriana, Bogota - Colombia / Estudio de muestras de fluidos corporales utilizando citometría de flujo: seis años de experiencia en el Hospital Universitario San Ignacio - Pontificia Universidad Javeriana Bogotá, Colombia / Estudo de amostras de fluídos corporais utilizando citometria de fluxo: seis anos de experiencia no Hospital Universitário San Ignacio - Pontificia Universidad Javeriana, Bogotá, Colômbia

Abstract Flow cytometry (FCM) was implemented in 2008 at the Pontificia Universidad Javeriana and later at the Hospital Universitario San Ignacio to examine special samples of patients with hematological malignancies and solid tumors other than bone marrow and peripheral blood for diagnosis and monitoring. This study describes the main findings of special sample evaluation over a six-year period. In all, 1070 samples of body fluids from patients with benign and malignant diseases were examined by FCM. These samples were stabilized with TransFixTM and stained with six-color immunophenotyping panels. Samples included cerebrospinal fluid, bronchoalveolar lavage, pleural fluid, pericardial fluid and ascite fluid from patients with acute and chronic leukemia, myelodysplastic syndromes, lymphomas, myeloma, autoimmune diseases, immunodeficiencies and solid tumors, among others. Flow cytometry provided important information for the classification and detection of minimal numbers of tumor cells in leukemia and lymphoma cases. This work represents the first national report describing FCM implementation in special samples for diagnosis and clinical monitoring of patients with malignant and benign pathologies.

Resumen La citometría de flujo fue implementada en 2008 en la Pontificia Universidad Javeriana y posteriormente en el Hospital San Ignacio con el fin de examinar, para diagnóstico y monitoreo, muestras especiales de pacientes con malignidades hematológicas y tumores sólidos, distintos de los de médula ósea y sangre periférica. Este estudio describe los principales hallazgos de la evaluación de estas muestras especiales en un periodo de seis años. En total, se examinaron por citometría de flujo 1070 muestras de fluidos corporales de pacientes con enfermedades malignas y benignas. Estas muestras se estabilizaron con TransFix™ y teñidas con paneles inmunofenotípicos de seis colores. Las muestras incluyeron líquido cefalorraquídeo, lavado broncoalveolar, fluido pleural, fluido pericárdico y fluido ascítico, provenientes de pacientes con leucemia aguda y crónica, síndromes mielodisplásicos, linfomas, mieloma, enfermedades autoinmunes, inmunodeficiencias y tumores sólidos, entre otras enfermedades. La citometría de flujo proporcionó información importante sobre la clasificación y detección de números mínimos de células tumorales en casos de leucemia y linfoma. Este trabajo representa el primer reporte nacional que describe la implementación de citometría de flujo en muestras especiales para diagnóstico y monitoreo clínico de pacientes con patologías malignas y benignas.

Resumo A citometria de fluxo foi implementada em 2008 na Pontificia Universidad Javeriana e posteriormente no Hospital San Ignacio com a finalidade de examinar amostras especiais de pacientes com malignidades hematológicas e tumores sólidos diferentes aos de medula óssea e sangue periférico para diagnóstico e monitoramento. Este estudo descreve as principais descobertas a partir da avaliação de amostras especiais em um período de 6 anos. Se examinaram por citometria de fluxo um total de 1.070 amostras de fluídos corporais de pacientes doenças malignas e benignas. Estas amostras se estabilizaram com TransFix™ e coradas com painéis imunofenotípicos de seis cores. As amostras incluíram fluído cérebro-espinhal, lavado broncoalveolar, fluído pleural, fluído pericárdico e fluído ascético, provenientes de pacientes com leucemias aguda e crónica, síndromes mielodisplásicos, linfomas, mielomas, doenças autoimunes, imunodeficiência, e tumores sólidos, entre outras doenças. A citometria de fluxo proporcionou informações importantes sobre a classificação e detecção de um número mínimo de células tumorais nos casos de leucemia e linfomas. Este trabalho representa o primeiro relato nacional que descreve a implementação de citometria de fluxo em amostras especiais para diagnóstico e monitoramento clínico de pacientes com patologias malignas e benignas.

Prognostic stratification improvement by integrating ID1/ID3/IGJ gene expression signature and immunophenotypic profile in adult patients with B-ALL.

BACKGROUND: Survival of adults with B-Acute Lymphoblastic Leukemia requires accurate risk stratification of patients in order to provide the appropriate therapy. Contemporary techniques, using clinical and cytogenetic variables are incomplete for prognosis prediction. METHODS: To improve the classification of adult patients diagnosed with B-ALL into prognosis groups, two strategies were examined and combined: the expression of the ID1/ID3/IGJ gene signature by RT-PCR and the immunophenotypic profile of 19 markers proposed in the EuroFlow protocol by Flow Cytometry in bone marrow samples. RESULTS: Both techniques were correlated to stratify patients into prognostic groups. An inverse relationship between survival and expression of the three-genes signature was observed and an immunophenotypic profile associated with clinical outcome was identified. Markers CD10 and CD20 were correlated with simultaneous overexpression of ID1, ID3 and IGJ. Patients with simultaneous expression of the poor prognosis gene signature and overexpression of CD10 or CD20, had worse Event Free Survival and Overall Survival than patients who had either the poor prognosis gene expression signature or only CD20 or CD10 overexpressed. CONCLUSION: By utilizing the combined evaluation of these two immunophenotypic markers along with the poor prognosis gene expression signature, the risk stratification can be significantly strengthened. Further studies including a large number of patients are needed to confirm these findings.

High expression of ID family and IGJ genes signature as predictor of low induction treatment response and worst survival in adult Hispanic patients with B-acute lymphoblastic leukemia.

BACKGROUND: B-Acute lymphoblastic leukemia (B-ALL) represents a hematologic malignancy with poor clinical outcome and low survival rates in adult patients. Remission rates in Hispanic population are almost 30% lower and Overall Survival (OS) nearly two years inferior than those reported in other ethnic groups. Only 61% of Colombian adult patients with ALL achieve complete remission (CR), median overall survival is 11.3 months and event-free survival (EFS) is 7.34 months. Identification of prognostic factors is crucial for the application of proper treatment strategies and subsequently for successful outcome. Our goal was to identify a gene expression signature that might correlate with response to therapy and evaluate the utility of these as prognostic tool in hispanic patients. METHODS: We included 43 adult patients newly diagnosed with B-ALL. We used microarray analysis in order to identify genes that distinguish poor from good response to treatment using differential gene expression analysis. The expression profile was validated by real-time PCR (RT-PCT). RESULTS: We identified 442 differentially expressed genes between responders and non-responders to induction treatment. Hierarchical analysis according to the expression of a 7-gene signature revealed 2 subsets of patients that differed in their clinical characteristics and outcome. CONCLUSIONS: Our study suggests that response to induction treatment and clinical outcome of Hispanic patients can be predicted from the onset of the disease and that gene expression profiles can be used to stratify patient risk adequately and accurately. The present study represents the first that shows the gene expression profiling of B-ALL Colombian adults and its relevance for stratification in the early course of disease.