Mutations in LRRK2 impair NF-κB pathway in iPSC-derived neurons.

BACKGROUND: Mutations in leucine-rich repeat kinase 2 (LRRK2) contribute to both familial and idiopathic forms of Parkinson's disease (PD). Neuroinflammation is a key event in neurodegeneration and aging, and there is mounting evidence of LRRK2 involvement in inflammatory pathways. In a previous study, we described an alteration of the inflammatory response in dermal fibroblasts from PD patients expressing the G2019S and R1441G mutations in LRRK2. METHODS: Taking advantage of cellular reprogramming, we generated induced pluripotent stem cell (iPSC) lines and neurons thereafter, harboring LRRK2G2019S and LRRK2R1441G mutations. We used gene silencing and functional reporter assays to characterize the effect of the mutations. We examined the temporal profile of TNFα-induced changes in proteins of the NF-κB pathway and optimized western blot analysis to capture α-synuclein dynamics. The effects of the mutations and interventions were analyzed by two-way ANOVA tests with respect to corresponding controls. RESULTS: LRRK2 silencing decreased α-synuclein protein levels in mutated neurons and modified NF-κB transcriptional targets, such as PTGS2 (COX-2) and TNFAIP3 (A20). We next tested whether NF-κB and α-synuclein pathways converged and found that TNFα modulated α-synuclein levels, although we could not detect an effect of LRRK2 mutations, partly because of the individual variability. Nevertheless, we confirmed NF-κB dysregulation in mutated neurons, as shown by a protracted recovery of IκBα and a clear impairment in p65 nuclear translocation in the LRRK2 mutants. CONCLUSIONS: Altogether, our results show that LRRK2 mutations affect α-synuclein regulation and impair NF-κB canonical signaling in iPSC-derived neurons. TNFα modulated α-synuclein proteostasis but was not modified by the LRRK2 mutations in this paradigm. These results strengthen the link between LRRK2 and the innate immunity system underscoring the involvement of inflammatory pathways in the neurodegenerative process in PD.

Substance P, NPY, CCK and their receptors in five brain regions in major depressive disorder with transcriptomic analysis of locus coeruleus neurons.

Major depressive disorder (MDD) is a serious disease and a burden to patients, families and society. Rodent experiments and human studies suggest that several neuropeptide systems are involved in mood regulation. The aim of this study is two-fold: (i) to monitor, with qPCR, transcript levels of the substance P/tachykinin (TAC), NPY and CCK systems in bulk samples from control and suicide subjects, targeting five postmortem brain regions including locus coeruleus (LC); and (ii) to analyse expression of neuropeptide family transcripts in LC neurons of 'normal' postmortem brains by using laser capture microdissection with Smart-Seq2 RNA sequencing. qPCR revealed distinct regional expression patterns in male and female controls with higher levels for the TAC system in the dorsal raphe nucleus and LC, versus higher transcripts levels of the NPY and CCK systems in prefrontal cortex. In suicide patients, TAC, TAC receptors and a few NPY family transcript levels were increased mainly in prefrontal cortex and LC. The second study on 'normal' noradrenergic LC neurons revealed expression of transcripts for GAL, NPY, TAC1, CCK, and TACR1 and many other peptides (e.g. Cerebellin4 and CARTPT) and receptors (e.g. Adcyap1R1 and GPR173). These data and our previous results on suicide brains indicates that the tachykinin and galanin systems may be valid targets for developing antidepressant medicines. Moreover, the perturbation of neuropeptide systems in MDD patients, and the detection of further neuropeptide and receptor transcripts in LC, shed new light on signalling in noradrenergic LC neurons and on mechanisms possibly associated with mood disorders.

Spatial RNA Sequencing Identifies Robust Markers of Vulnerable and Resistant Human Midbrain Dopamine Neurons and Their Expression in Parkinson's Disease.

Defining transcriptional profiles of substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) dopamine neurons is critical to understanding their differential vulnerability in Parkinson's Disease (PD). Here, we determine transcriptomes of human SNc and VTA dopamine neurons using LCM-seq on a large sample cohort. We apply a bootstrapping strategy as sample input to DESeq2 and identify 33 stably differentially expressed genes (DEGs) between these two subpopulations. We also compute a minimal sample size for identification of stable DEGs, which highlights why previous reported profiles from small sample sizes display extensive variability. Network analysis reveal gene interactions unique to each subpopulation and highlight differences in regulation of mitochondrial stability, apoptosis, neuronal survival, cytoskeleton regulation, extracellular matrix modulation as well as synapse integrity, which could explain the relative resilience of VTA dopamine neurons. Analysis of PD tissues showed that while identified stable DEGs can distinguish the subpopulations also in disease, the SNc markers SLIT1 and ATP2A3 were down-regulated and thus appears to be biomarkers of disease. In summary, our study identifies human SNc and VTA marker profiles, which will be instrumental for studies aiming to modulate dopamine neuron resilience and to validate cell identity of stem cell-derived dopamine neurons.

Axon-seq for in Depth Analysis of the RNA Content of Neuronal Processes.

Neuronal processes have an RNA composition that is distinct from the cell body. Therefore, to fully understand neuronal biology in health and disease we need to study both somas, dendrites and axons. Here we describe a detailed protocol of a newly refined method, Axon-seq, for RNA sequencing of axons (and dendrites) grown in isolation using single microfluidic devices. We also detail how to generate motor neurons from mouse and human pluripotent stem cells for sequencing, but Axon-seq is applicable to any neuronal cell. In Axon-seq, the axons are recruited through a growth factor gradient, lysed and directly processed to cDNA without RNA isolation. A careful bioinformatic step ensures that any soma-contaminated samples are easily identified and removed.

CARL y MPL en pacientes cubanos con trombocitemia esencial y mielofibrosis primaria

Introducción: La trombocitemia esencial y la mielofibrosis primaria comparten la presencia de las mutaciones JAK2, CALR y MPL. En total, están presentes en poco más del 90 % de los pacientes con estas enfermedades. Objetivos: Determinar el comportamiento de las mutaciones más frecuentes en los genes MPL y CALR en pacientes cubanos. Métodos: Se realizó un estudio ambispectivo, descriptivo y longitudinal en el Instituto de Hematología e Inmunología de Cuba, entre los años 2010 y 2020. Se incluyeron todos los pacientes con sospecha de trombocitemia esencial y de mielofibrosis primaria con muestras de ADN válidas. Se les identificaron las mutaciones CALR y MPL por PCR en tiempo real. Resultados: De los 53 pacientes estudiados, el 67,9 % fueron diagnosticados con trombocitemia esencial, el 22,6 % con mielofibrosis primaria. En el 90,6 % se pudo detectar alguna de las mutaciones conductoras; el 67,9 % fueron positivos a la mutación JAK2V617F, el 13,2 % a las mutaciones en el gen que codifica para la calreticulina y en el 9,4 % se identificaron mutaciones en el gen MPL. Conclusiones: El comportamiento de las mutaciones conductoras JAK2V617F, CALR y MPL en la muestra de pacientes cubanos con trombocitemia esencial y mielofibrosis primaria estuvo en correspondencia con lo descrito en la mayoría de las investigaciones.

Introduction: Essential thrombocythemia and primary myelofibrosis share the presence of JAK2, CALR and MPL mutations. In total, they comprise slightly more than 90 % of patients with these diseases. Objectives: To determine the behavior of the most frequent mutations in MPL and CALR genes in Cuban patients. Methods: An ambispective, descriptive and longitudinal study was performed at the Institute of Hematology and Immunology of Cuba, between 2010 and 2020. All patients with suspected essential thrombocythemia and primary myelofibrosis with valid DNA samples were included. CALR and MPL mutations were identified by real-time PCR. Results: Of the 53 patients studied, 67.9% were diagnosed with essential thrombocythemia, and 22.6% with primary myelofibrosis. In 90.6% it was possible to detect any of the driver mutations: 67.9% were positive for the JAK2V617F mutation, 13.2% for mutations in the gene coding for calreticulin and in 9.4% mutations in the MPL gene were identified. Conclusions: The behavior of the driver mutations JAK2V617F, CALR and MPL in the sample of Cuban patients with essential thrombocythemia and primary myelofibrosis was in correspondence with what is described in the majority of the investigations.

Axon-Seq Decodes the Motor Axon Transcriptome and Its Modulation in Response to ALS.

Spinal motor axons traverse large distances to innervate target muscles, thus requiring local control of cellular events for proper functioning. To interrogate axon-specific processes we developed Axon-seq, a refined method incorporating microfluidics, RNA sequencing (RNA-seq), and bioinformatic quality control. We show that the axonal transcriptome is distinct from that of somas and contains fewer genes. We identified 3,500-5,000 transcripts in mouse and human stem cell-derived spinal motor axons, most of which are required for oxidative energy production and ribogenesis. Axons contained transcription factor mRNAs, e.g., Ybx1, with implications for local functions. As motor axons degenerate in amyotrophic lateral sclerosis (ALS), we investigated their response to the SOD1G93A mutation, identifying 121 ALS-dysregulated transcripts. Several of these are implicated in axonal function, including Nrp1, Dbn1, and Nek1, a known ALS-causing gene. In conclusion, Axon-seq provides an improved method for RNA-seq of axons, increasing our understanding of peripheral axon biology and identifying therapeutic targets in motor neuron disease.

Niveles de β2microglobulina, variables clínicas y sociodemográficas en pacientes con síndrome linfoproliferativo crónico de reciente diagnóstico

Fundamento: la β2microglobulina está reconocida como marcador tumoral para diferentes propósitos en hematopatías malignas de estirpe linfoide; sin embargo, no hay antecedentes de su utilización en la provincia de Cienfuegos. Objetivo describir las características sociodemográficas, clínicas y la distribución de los niveles séricos de β2microglobulina en pacientes con síndrome linfoproliferativo crónico y su relación con los estadios clínicos y la respuesta al tratamiento de primera línea. Métodos: estudio observacional descriptivo transversal. La serie se conformó con todos los pacientes adultos con diagnóstico reciente (sin comenzar terapia antitumoral específica) de mieloma múltiple, leucemia linfoide crónica, linfoma no Hodgkin y linfoma Hodgkin, ingresados en el Servicio de Hematología del Hospital General Universitario Dr. Gustavo Aldereguìa Lima, durante el año 2020. La información se obtuvo mediante revisión documental de historias clínicas y ensayos de laboratorio. Se analizaron las variables: sexo, edad, color de la piel, niveles de β2microglobulina, tipo de enfermedad, estadios clínicos y respuesta al tratamiento. Resultados: el 84 % de la serie presentó niveles elevados del analito, más acentuado en el mieloma. Se constató relación entre los niveles estratificados de β2microglobulina con los estadios clínicos y la respuesta al tratamiento de primera línea. Conclusiones: las características sociodemográficas y las variables clínicas observadas no difieren de forma sustantiva con lo reportado. La distribución de los niveles de la β2microglobulina es sugerente de una relación directa entre los estadios clínicos e inversa con la respuesta al tratamiento.

Background: β2microglobulin is recognized as a tumor marker for different purposes in malignant hematopathies of lymphoid lineage; however, there is no history of its use in the Cienfuegos province. Objective: to describe the sociodemographic and clinical characteristics and the distribution of serum β2microglobulin levels in patients with chronic lymphoproliferative syndrome and their relationship with clinical stages and response to first-line treatment. Methods: cross-sectional descriptive observational study. The series was made up of all adult patients (universe 50) recently diagnosed (without starting specific antitumor therapy) of multiple myeloma, chronic lymphoid leukemia, non-Hodgkin lymphoma and Hodgkin lymphoma, admitted to the Hematology Service of the Dr. Gustavo Aldereguìa Lima General University Hospital, during the year 2020. The information was obtained through documentary review of medical records and laboratory tests. The analyzed variables were: sex, age, skin color, β2microglobulin levels, type of disease, clinical stages and response to treatment. Results: 84% of the series presented high levels of the analyte, more accentuated in myeloma. A relationship was found between the stratified levels of β2microglobulin with the clinical stages and the response to first-line treatment. Conclusions: the sociodemographic characteristics and the clinical variables observed do not differ substantially from what was reported. The distribution of β2microglobulin levels is suggestive of a direct relationship between clinical stages and an inverse relationship with response to treatment.

Resultados del trasplante autólogo de progenitores hematopoyéticos en pacientes con mieloma múltiple / Outcome of autologous stem cell transplantation in patients with multiple myeloma

Introducción: El mieloma múltiple es una neoplasia caracterizada por la proliferación de un clon de células plasmáticas monoclonales. Representa el 1 por ciento de todos los cánceres y el 10 por ciento de las neoplasias hematológicas. Las altas dosis de quimioterapia seguidas de trasplante autólogo de progenitores hematopoyéticos constituyen una opción terapéutica segura para un grupo seleccionado de pacientes. Objetivo: Analizar los resultados del trasplante autólogo de progenitores hematopoyéticos en pacientes con mieloma múltiple. Métodos: Estudio descriptivo, longitudinal, ambispectivo. El universo estuvo conformado por 14 pacientes (de los cuales se incluyeron 13) con diagnóstico de mieloma múltiple tratados con trasplante autólogo de progenitores hematopoyéticos de sangre periférica como tratamiento de consolidación, en el Instituto de Hematología e Inmunología de La Habana, desde enero de 2014 hasta diciembre de 2019. Resultados: La edad media fue de 53,9±5,6 años, predominó el sexo femenino. Las complicaciones más frecuentes fueron las infecciosas y la mucositis. La supervivencia global al año fue del 100 por ciento y del 80 por ciento a los cinco años. La supervivencia libre de progresión al año fue de 83 por ciento y 73 por ciento a los cinco años. Conclusiones: El trasplante hematopoyético autólogo de sangre periférica en pacientes con mieloma múltiple es un tratamiento que eleva las tasas de respuestas, la supervivencia global y libre de progresión y generalmente presenta pocas complicaciones(AU)

Introduction: Multiple myeloma is a neoplasm characterized by the presence of a plasmatic cells clone; accounts for 1 percent of cancers, and approximately 10 percent of hematologic malignancies. High dose chemotherapy followed by autologous stem cell transplantation remains a safe option in selected patients. Objective: To analyze the results of autologous stem cell transplantation from peripheral blood in patients with multiple myeloma. Methods: A descriptive, longitudinal, ambispective study. The universe was by 14 patients (13 of them were finally included), with diagnosis of multiple myeloma treated with autologous stem cell transplantation from peripheral blood at Institute of Hematology and Immunology of Havana from January 2014 to December 2019. Results: The median age of the patient was 53.9±5.6 years, with predominance of female sex. More frequents complications were infections and mucositis. A year overall survival was 100 percent and 80 percent at five years. Progression free survival at one and five years was 83 percent and 73 percent, respectively. Conclusions: The autologous stem cell transplantation in multiple myeloma patients increase complete responses, overall survival and progression free survival and is well accepted without severe complications(AU)

Disfunción crónica de órganos en pacientes con drepanocitosis. Parte I: manifestaciones cardiorrespiratorias / Chronic organ dysfunction in patients with sickle cell anemia. Part I: cardiorespiratory manifestations

Introducción: En la actualidad la mayoría de los pacientes con anemia falciforme alcanzan la adultez. La disfunción crónica de órganos constituye la causa primaria de muerte y representa un desafío en el manejo de estos enfermos. Objetivo: Analizar los aspectos generales de la disfunción orgánica crónica en pacientes con drepanocitosis y profundizar en las alteraciones cardiorrespiratorias. Métodos: Se realizó una revisión de los artículos publicados en los últimos diez años con el uso de los buscadores PubMed, SciELO y Google Académico. Los términos de búsqueda fueron: anemia de células falciformes, disfunción orgánica, mortalidad, hipertensión pulmonar, enfermedad pulmonar crónica, asma, apnea obstructiva del sueño. Análisis y síntesis de la información: Con el aumento de la expectativa de vida, en el manejo de los pacientes con drepanocitosis, gana en relevancia el diagnóstico precoz y el tratamiento oportuno de la disfunción crónica de órganos. Este es un proceso que comienza en la infancia, pero se hace más evidente en la etapa adulta. Las manifestaciones cardiorrespiratorias más comunes y con impacto en la morbilidad y en la mortalidad son: la hipertensión pulmonar y la enfermedad pulmonar crónica. Se describen aspectos relacionado con la prevalencia, diagnóstico, implicaciones en la evolución de los enfermos y aspectos relacionados con su tratamiento. Conclusiones: La hipertensión pulmonar y la enfermedad pulmonar crónica son frecuentes en pacientes con drepanocitosis. Ambas complicaciones tienen un impacto negativo en la evolución de estos enfermos y se asocian a aumento de la mortalidad. La detección temprana de estas afecciones, permite tomar acciones terapéuticas para disminuir sus consecuencias(AU)

Introduction: Currently, most patients with sickle cell disease reach adulthood. Chronic organ dysfunction constitutes the primary cause of death and represents a challenge in the management of these patients. Objectives: To analyze the general aspects of chronic organic dysfunction in patients with sickle cell disease and to delve into the cardiorespiratory ones. Methods: A review of the articles published in the last ten years was carried out using the PubMed, SciELO and Google Scholar search engines. The search terms were: sickle cell anemia, organ dysfunction, mortality, pulmonary hypertension, chronic lung disease, asthma, obstructive sleep apnea. Information analysis and synthesis: With the increase in life expectancy, in the management of patients with sickle cell disease, early diagnosis and timely treatment of chronic organ dysfunction gain relevance. This is a process that begins in childhood, but becomes more apparent in adulthood. The most common cardiorespiratory manifestations with an impact on morbidity and mortality are: pulmonary hypertension and chronic lung disease. Aspects related to prevalence, diagnosis, implications in the evolution of patients and aspects related to their treatment are described. Conclusions: Pulmonary hypertension and chronic lung disease are frequent in patients with sickle cell disease. Both complications have a negative impact on the evolution of these patients and are associated with increased mortality. Early detection of these conditions allows therapeutic actions to be taken to reduce their consequences(AU)

Estimación del filtrado glomerular por valores séricos de creatinina y cistatina C en adultos con drepanocitosis / Estimation of glomerular filtration rate using serum creatinine and cystatin C in adults with sickle cell anemia

Introducción: La estimación del filtrado glomerular a partir de la concentración sérica de creatinina en pacientes con drepanocitosis, es anormal cuando hay un deterioro extenso de la función renal. Objetivos: Evaluar la estimación del filtrado glomerular con el uso de creatinina, de cistatina C y de ambas determinaciones, en adultos con drepanocitosis. Métodos: Estudio observacional, descriptivo de corte transversal. Se incluyeron 44 adultos con drepanocitosis en quienes se estimó el filtrado glomerular, con el uso de creatinina, de cistatina C y de ambas, según las fórmulas de CDK-EPI. Resultados: Se encontraron discrepancias en el filtrado glomerular estimado por creatinina, por cistatina C y por ambas (medias: 112,2 ± 28,4; 55,7 ± 23,1 y 75,1 ± 24,7 mL/min/ 1,73 m2; respectivamente). Los porcentajes más elevados de pacientes con hiperfiltración por creatinina corresponden al genotipo SS y todos los que tiene enfermedad renal crónica en estadio 4 (filtrado estimado por cistatina C y mediante la combinación de ambos marcadores), tienen este tipo de hemoglobinopatía. La hiperfiltración fue más común en los más jóvenes y la disminución del filtrado en los mayores de 40 años. Conclusiones: La estimación del filtrado glomerular muestra diferencias entre los tres métodos estudiados. Con el uso de creatinina está sobrestimado y una elevada proporción de pacientes son clasificados como con hiperfiltración. El uso de la cistatina C o la combinación de ambas determinaciones, pudieran ofrecer una estimación más exacta del filtrado glomerular en pacientes con drepanocitosis(AU)

Introduction: The estimation of the glomerular filtration rate from the serum concentration of creatinine in patients with sickle cell disease is abnormal when there is an extensive deterioration of renal function. Objective: To estimate the glomerular filtration rate with the use of creatinine, cystatin C and both determinations, in adults with sickle cell disease. Determine if there are differences between these methods. Methods: Observational, descriptive cross-sectional study. Forty-four adults with sickle cell disease were included in whom the glomerular filtration rate was estimated using creatinine, cystatin C and both, according to the CDK-EPI formulas. Results: Discrepancies were found in the glomerular filtration rate estimated by creatinine, by cystatin C and by both (means: 112.2 ± 28.4; 55.7 ± 23.1 and 75.1 ± 24.7 mL/min/1.73 m2; respectively). The highest percentages of patients with creatinine hyperfiltration correspond to the SS genotype and all those with stage 4 chronic kidney disease (estimated filtration by cystatin C and by the combination of both markers), have this type of hemoglobinopathy. Hyperfiltration was more common in the youngest and decreased filtration in those older than 40 years. Conclusions: The estimation of the glomerular filtration rate shows differences between the three methods studied. With the use of creatinine, it is overestimated and a high proportion of patients are classified as having hyperfiltration. The use of cystatin C or the combination of both determinations could offer a more accurate estimate of glomerular filtration rate in patients with sickle cell disease(AU)

Influencia de la carga alélica del JAK2V617F en pacientes cubanos con trombocitemia esencial y mielofibrosis primaria / Influence of JAK2V617F allelic burden in Cuban patients with essential thrombocythemia and primary myelofibrosis

Introducción: La frecuencia de la mutación JAK2V617F se estima entre el 50 y 60 por ciento en pacientes con trombocitemia esencial y mielofibrosis primaria. El 30 por ciento de los pacientes con policitemia vera y mielofibrosis primaria. Entre 2-4 por ciento de los pacientes con trombocitemia esencial presentan pérdida de heterocigosidad. Objetivos: Evaluar la influencia de la carga alélica de la mutación JAK2V617F y su relación con variables clínico-hematológicas en el diagnóstico de estas enfermedades en pacientes cubanos. Métodos: Se realizó un estudio retrospectivo, descriptivo y longitudinal en el Instituto de Hematología e Inmunología entre 2010 y 2020. Se incluyeron todos los pacientes con sospecha de trombocitemia esencial y mielofibrosis primaria con muestras de ADN válidas. Se les cuantificó la carga alélica de la mutación por PCR en tiempo real. Resultados: Se detectó la mutación en 66,7 por ciento de los diagnosticados con trombocitemia esencial y mielofibrosis primaria. El 62,5 por ciento de los pacientes con mielofibrosis primaria fueron homocigotos a la mutación, mientras que en la trombocitemia esencial solo el 20,8 por ciento. La diferencia de medias de cargas alélicas entre ambas enfermedades fue estadísticamente significativa. No se encontraron diferencias significativas en la comparación de las variables clínicas y hematológicas en estas enfermedades ni asociación con la carga alélica con excepción de las plaquetas en la mielofibrosis primaria. Conclusiones: El estudio estuvo limitado por la escasa muestra de pacientes, pero se corresponde con otras investigaciones que sostienen el concepto de que la presentación fenotípica de las neoplasias mieloproliferativasestá influenciada por la carga mutacional del JAK2V617F(AU)

Introduction: The frequency of the JAK2V617F mutation is estimated to be between 50 percent and 60 percent in patients with essential thrombocythemia and primary myelofibrosis. 30 percent of patients with polycythemia vera and primary myelofibrosis and 2-4 percent of patients with essential thrombocythemia show loss of heterozygosity. Objectives: To evaluate the influence of the allelic load of the JAK2V617F mutation in the diagnosis of these diseases in Cuban patients and its relationship with clinical-hematological variables. Methodology: A retrospective, descriptive and longitudinal study was carried out at the Institute of Hematology and Immunology between 2010 and 2020. All patients with suspected essential thrombocythemia and primary myelofibrosis with valid DNA samples were included. The allelic load of the mutation was quantified by real-time PCR. Results: The mutation was detected in 66.7 percent of those diagnosed with essential thrombocythemia and primary myelofibrosis. 62.5 percent of the patients with primary myelofibrosis were homozygous for the mutation, while in essential thrombocythemia only 20.8 percent. The difference in mean allelic loads between both diseases was statistically significant. No significant differences were found in the comparison of clinical and hematological variables in these diseases or association with allelic load, with the exception of platelets in primary myelofibrosis. Conclusions: The study was limited by the small sample of patients, but it corresponds to other investigations that support the concept that the phenotypic presentation of myeloproliferative neoplasms is influenced by the mutational load of JAK2V617F(AU)

Identificación del riesgo de trombosis en pacientes con padecimientos oncológicos ambulatorios en quimioterapia

RESUMEN Fundamento existe una relación establecida entre el cáncer y la aparición y desarrollo de enfermedad tromboembólica. Se hace necesario identificar a los pacientes con un riesgo aumentado de tromboembolismo venoso utilizando escalas predictivas para individualizar el riesgo. Objetivo identificar el riesgo de trombosis en el paciente con padecimientos oncológicos ambulatorios en quimioterapia. Método se realizó un estudio descriptivo de corte transversal, en una serie de casos, atendidos en el Servicio de Oncología Clínica del Hospital General Universitario Dr. Gustavo Aldereguía Lima, de Cienfuegos, en el período comprendido entre el primero de junio de 2020 hasta el treinta de abril de 2021. La información se obtuvo del expediente clínico y un modelo recolector de datos. Las variables analizadas fueron: edad, sexo, sitio del tumor primario, conteo de plaquetas previo a la quimioterapia, niveles de hemoglobina sérica, uso de eritropoyetina, conteo leucocitario previo a la quimioterapia e índice de masa corporal. Los datos obtenidos se procesaron mediante el programa estadístico SPSS versión 21 y los resultados se presentan en tablas mediante números absolutos y porcientos. Resultados el grupo de edad más frecuente fue el de 60 años y más con 77 pacientes (51,3 %), del sexo masculino 45 (30 %). La aplicación del modelo de predicción de Khorana marcó un porcentaje relevante (26 %) de pacientes con riesgo intermedio y alto de trombosis. Conclusiones la investigación muestra una población adulta mayoritariamente envejecida, con factores de riesgo de trombosis relacionados con el paciente, la enfermedad oncológica y los tratamientos sistémicos.

ABSTRACT Background: there is an established relationship between cancer and the appearance and development of thromboembolic disease. It is necessary to identify patients with an increased risk of venous thromboembolism using predictive scales to individualize the risk. Objective: to identify the risk of thrombosis in patients with suffering cancer conditions undergoing chemotherapy. Method: a series of cases descriptive cross-sectional study, treated at the Clinical Oncology Service of the Dr. Gustavo Aldereguía Lima University General Hospital, Cienfuegos, from June the 1st, 2020 to April 30th, 2021. The information was obtained from the clinical record and a data collection model. The variables analyzed were: age, sex, site of the primary tumor, platelet count prior to chemotherapy, serum hemoglobin levels, erythropoietin use, leukocyte count prior to chemotherapy, and body mass index. The data obtained were processed using the SPSS version 21 statistical program and the results are presented in tables using absolute numbers and percentages. Results: the most frequent age group was 60 years old and over with 77 patients (51.3 %), 45 (30 %) male. The application of the Khorana score marked a relevant percentage (26 %) of patients with intermediate and high risk of thrombosis. Conclusions: the research shows a mostly aged adult population, with risk factors for thrombosis related to the patient, oncological disease and systemic treatments.

Capacitación en ensayos clínicos. Cienfuegos 2004-2018

RESUMEN Fundamento: los profesionales de la salud deben estar adecuadamente calificados para desempeñar sus funciones como investigadores. En Cienfuegos se han impartido varios cursos para capacitar al personal en ensayos clínicos, lo cual permite elevar la calidad de este tipo de investigación. Objetivo: describir el comportamiento de la capacitación del personal de salud en la temática de ensayos clínicos durante el periodo desde 2004 hasta 2018 en la provincia Cienfuegos. Métodos: estudio descriptivo sobre los cursos impartidos en Cienfuegos en el periodo comprendido desde 2004 hasta 2018. Se consultó la documentación legislada por secretaria docente de postgrado para la impartición de cursos. Se analizaron las variables: cursos impartidos, años en que se impartieron, investigadores capacitados, lugar de procedencia y profesión. Resultados: se impartieron 46 cursos, de ellos 19 para la capacitación de investigadores clínicos. Se capacitaron 366 investigadores, de los cuales 283 (77,3 %) pertenecen a la atención secundaria y 83 (22,7 %) a la atención primaria de salud. Entre los años 2013 y 2017 se alcanzaron las mayores cifras: 30 cursos y 163 investigadores capacitados para un el 46, 2 %. Según su profesión, el 52, 5 % son médicos y el 21,9 % son enfermeros. Conclusiones: en Cienfuegos se capacitó al personal que participó en la ejecución de ensayos clínicos; con mayor representación en el personal médico y de enfermería de la atención secundaria en relación al número de ensayos clínicos que se ejecutan en este nivel del sistema de salud.

ABSTRACT Background: Health professionals must be adequately qualified to perform their duties as investigators. In Cienfuegos, several courses have been given to train staff in clinical trials, which allows raising the quality of this type of research. Objective: Describe the behavior of the training of health personnel on the subject of clinical trials in the period from 2004 to 2018 in the province of Cienfuegos. Methods: Descriptive study on the courses taught in Cienfuegos in the period from 2004 to 2018. The documentation legislated by the postgraduate teaching secretary for the delivery of courses was consulted. The variables were analyzed: courses taught, years in which they were taught, researchers trained, place of origin and profession. Results: 46 courses were given, of them 19 for the training of clinical investigators. 366 researchers were trained, of which 283 (77.3%) belong to secondary care and 83 (22.7%) to primary health care. In the study period, it was identified that between 2013 and 2017 higher figures were reached, 30 courses and 163 trained researchers for 46.2%. According to their profession, 52.5% are doctors and 21.9% are nurses. Conclusions: In Cienfuegos, the personnel who participated in the execution of clinical trials were trained; with greater representation in the medical and nursing staff of secondary care in relation to the number of clinical trials carried out at this level of the health system.

Alteraciones funcionales ventilatorias en pacientes adultos con drepanocitosis / Functional ventilatory alterations in adult patients with sickle cell disease

RESUMEN Fundamento: en los pacientes con drepanocitosis las alteraciones de la función pulmonar son frecuentes y pueden detectarse por la espirometría. Objetivo: describir las alteraciones de la función ventilatoria en los pacientes con drepanocitosis seguidos en consulta provincial de hemoglobinopatías. Métodos: estudio observacional, descriptivo, de corte transversal. Se incluyeron 76 pacientes, seleccionados por muestreo no probabilístico accidental, de 88 adultos seguidos en el Hospital Dr. Gustavo Aldereguía Lima. Las variables estudiadas fueron: edad, sexo, tipo de hemoglobinopatía, síntomas respiratorios, antecedentes de síndrome torácico agudo, (formas clínicas y número de eventos), comorbilidades y patrón ventilatorio. Las pruebas funcionales ventilatorias se realizaron con Microspiro digital Hellige. Resultados: el grupo de edad entre 20-29 años fue el más frecuente (34,2 %). Hubo discreto predominio del sexo femenino (56,1 %) y del genotipo SS (67,1 %). El más frecuente de los síntomas respiratorios fue la disnea y entre las comorbilidades la insuficiencia cardíaca en 9,2 % de los enfermos. En 42,1 % se registró antecedentes de síndrome torácico agudo, neumonía en la mayoría de los casos. Nueve enfermos (11,8 %) presentaron trastornos obstructivos y 43 enfermos (56,6 %) un patrón restrictivo.En el genotipo SS el riesgo de tener alteraciones en la espirometría resultó tres veces mayor que el resto [OR=3,2 IC (1,7-5,9)] Conclusiones: resultó común el hallazgo de prueba funcional ventilatoria alterada con predominio del patrón restrictivo. Estas modificaciones se observaron con mayor frecuencia en pacientes con el genotipo SS.

ABSTRACT Foundation: in patients with sickle cell disease, lung function abnormalities are frequent and can be detected by spirometry. Objective: to describe the disturbances of the respiratory functions in patients with sickle cell disease followed in a provincial consultation of hemoglobinopathies. Methods: observational, descriptive, cross-sectional study. Seventy six patients, selected by accidental non-probabilistic sampling, of 88 adults followed at the Dr. Gustavo Aldereguía Lima Hospital were included. The variables studied were: age, sex, type of hemoglobinopathy, respiratory symptoms, history of acute thoracic syndrome, (clinical forms and number of events), comorbidities and ventilatory pattern. Functional ventilatory tests were performed with Hellige digital Microspiro. Results: the age group between 20-29 years was the most frequent (34.2%). There was a discrete female predominance (56.1%) and the SS genotype (67.1%). The most frequent of the respiratory symptoms was dyspnea and among the comorbidities heart failure in 9.2% of the patients. In 42.1% there was a history of acute chest syndrome, pneumonia in most cases. Nine patients (11.8%) presented obstructive disorders and 43 patients (56.6%) a restrictive pattern. In the SS genotype the risk of having spirometry alterations was three times higher than the rest [OR = 3.2 IC (1.7-5.9)] Conclusion: the finding of altered respiratory functional test with a predominance of the restrictive pattern was common. These modifications were observed more frequently in patients with the SS genotype.

Linfoma linfocítico de células pequeñas de anexos oculares. Presentación de un caso / Small cell lymphocytic lymphoma of eye annexes. Case presentation

RESUMEN Los linfomas son enfermedades malignas que se originan en las células del sistema inmune y se manifiestan predominantemente por linfoadenopatías; en ocasiones la presentación es en sitios extranodales. El ojo y sus anexos pueden ser afectados por un grupo heterogéneo de linfomas primarios o, secundariamente, por la extensión de linfomas originados en ganglios linfáticos o en otros sitios extranodales. Se reporta el caso de un paciente que acudió a consulta por ptosis palpebral izquierda y al examen físico se detectó afectación bilateral, con presencia de tumores en la conjuntiva bulbar inferior en ojo derecho y en la conjuntiva bulbar superior en ojo izquierdo. Mediante biopsia de conjuntiva se diagnosticó linfoma no Hodgkin linfocítico de células pequeñas B. El paciente fue considerado un estadio IIIAE de Ann Arbor y tratado con seis ciclos de ciclofosfamida, vincristina, doxorrubicina y prednisona, más el anticuerpo monoclonal cubano anti-CD20 CIMABior®. Se alcanzó la respuesta completa, estado en el cual se mantiene hasta el momento.

ABSTRACT Lymphomas are malignant diseases that originate in the immune system cells and are predominantly manifested by lymphadenopathy; sometimes the presentation is in extranodal places. The eye and its annexes may be affected by a heterogeneous group of primary lymphomas or, secondarily, by the extension of lymphomas originated in lymph nodes or other extranodal sites. The case of a patient who went to the consultation due to left palpebral ptosis is reported and physical examination showed bilateral involvement, with tumors in the lower right eye bulbar conjunctiva and in the left eye upper bulbar conjunctiva. A small B cell lymphocytic non-Hodgkin lymphoma was diagnosed by conjunctiva biopsy. The patient was considered a stage IIIAE of Ann Arbor and treated with six cycles of cyclophosphamide, vincristine, doxorubicin and prednisone, plus the Cuban monoclonal antibody anti-CD20 CIMABior®. The complete response was reached, state in which the disease remains so far.

Leucine-rich repeat kinase 2 modulates cyclooxygenase 2 and the inflammatory response in idiopathic and genetic Parkinson's disease.

Inflammatory mechanisms are activated in aging and late-onset neurodegenerative diseases, such as Parkinson's disease (PD). Mutations in leucine-rich repeat kinase 2 (LRRK2) contribute to both idiopathic and familial forms of PD. Here, we investigated the involvement of LRRK2 in inflammatory pathways using primary dermal fibroblasts from patients with 2 common mutations in LRRK2 (G2019S and R1441G), idiopathic PD and age-matched healthy individuals. Basal cyclooxygenase (COX)-2 RNA levels were very high in the fibroblasts of all patients. Remarkably, LRRK2 silencing experiments significantly reduced basal COX-2 levels and COX-2 induction after a pro-inflammatory stimulus. Additionally, in samples from patients with the R1441G mutation and with idiopathic PD, we found a prominent cytoplasmic re-distribution of human antigen R, a protein that, among others, stabilizes COX-2 RNA. Furthermore, the response to lipopolysaccharide was defective in these 2 groups, which showed weak induction of pro-inflammatory cytokines and reduced NFκB transcriptional activation. In summary, we describe multiple defects in inflammatory pathways in which LRRK2 appears to be critically involved. Further studies are required to establish the therapeutic implications of inflammatory dysregulation in the pathophysiology of Parkinson's disease.

Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for Dopamine Neurons From Human Pluripotent Stem Cells.

Human embryonic and induced pluripotent stem cells are potential cell sources for regenerative approaches in Parkinson disease. Inductive differentiation protocols can generate midbrain dopamine neurons but result in heterogeneous cell mixtures. Therefore, selection strategies are necessary to obtain uniform dopamine cell populations. Here, we developed a selection approach using lentivirus vectors to express green fluorescent protein under the promoter region of FOXA2, a transcription factor that is expressed in the floor plate domain that gives rise to dopamine neurons during embryogenesis. We first validated the specificity of the vectors in human cell lines against a promoterless construct. We then selected FOXA2-positive neural progenitors from several human pluripotent stem cell lines, which demonstrated a gene expression profile typical for the ventral domain of the midbrain and floor plate, but failed to enrich for dopamine neurons. To investigate whether this was due to the selection approach, we overexpressed FOXA2 in neural progenitors derived from human pluripotent stem cell lines. FOXA2 forced expression resulted in an increased expression of floor plate but not mature neuronal markers. Furthermore, selection of the FOXA2 overexpressing fraction also failed to enrich for dopamine neurons. Collectively, our results suggest that FOXA2 is not sufficient to induce a dopaminergic fate in this system. On the other hand, our study demonstrates that a combined approach of promoter activation and lentivirus vector technology can be used as a versatile tool for the selection of a defined cell population from a variety of human pluripotent stem cell lines.

Linfoma primario del sistema nervioso central. Presentación de un caso / Primary Lymphoma of the Central Nervous System. Case Presentation

Los linfomas primarios del sistema nervioso central son una variedad poco común de linfoma no Hodgkin extranodal, que pueden afectar cerebro, médula espinal, ojos, meninges y nervios craneales. Se presenta el caso de una paciente de 51 años de edad, que acude a consulta por cefalea y se comprobó papiledema causado por lesión encefálica que requirió tratamiento quirúrgico. El estudio histológico permitió diagnosticar linfoma no hodgkiniano de alto grado de malignidad, de células B, CD20 positivo, con patrón tipo Burkitt periférico. La paciente recibió inmuno-quimioterapia y radioterapia, con evolución favorable. Por ser este tipo de tumor poco común y de comportamiento agresivo se decidió la presentación del caso.

Primary Lymphomas of the Central Nervous System are an uncommon variety of non- Hodgkin extra node which may affect the brain, spinal cord, eyes, meninges and cranial nerves. It is presented a case of a 51-year-old patient who came to the doctor complaining of headache and papilledema caused by an encephalic lesion was corroborated which required surgical treatment. Histological study allowed diagnosing a non-hodking lymphoma of a high malignancy, B cells, positive CD20, with a type of peripheral Burkitt pattern. The patient received immunochemo-therapy and radiotherapy, with a favorable evolution. Because it is an uncommon and aggressive tumor it was decided to present the case.

Cellular programming and reprogramming: sculpting cell fate for the production of dopamine neurons for cell therapy.

Pluripotent stem cells are regarded as a promising cell source to obtain human dopamine neurons in sufficient amounts and purity for cell replacement therapy. Importantly, the success of clinical applications depends on our ability to steer pluripotent stem cells towards the right neuronal identity. In Parkinson disease, the loss of dopamine neurons is more pronounced in the ventrolateral population that projects to the sensorimotor striatum. Because synapses are highly specific, only neurons with this precise identity will contribute, upon transplantation, to the synaptic reconstruction of the dorsal striatum. Thus, understanding the developmental cell program of the mesostriatal dopamine neurons is critical for the identification of the extrinsic signals and cell-intrinsic factors that instruct and, ultimately, determine cell identity. Here, we review how extrinsic signals and transcription factors act together during development to shape midbrain cell fates. Further, we discuss how these same factors can be applied in vitro to induce, select, and reprogram cells to the mesostriatal dopamine fate.