ABSTRACT
Pulmonary artery banding (PAB) is a procedure mainly performed during the neonatal period as an initial stage to definitive palliative reconstruction, a scenario in which the criteria for banding adjustment are well defined. However, the indication for BAP in the adult is extraordinarily rare, even more in patients with single ventricle and unrepaired transposition of the great arteries (TGA), and there are no established criteria for banding adjustment. Due to the small number of these procedures, there is limited experience in their anesthetic management and complications. We describe a case of a 29-year-old patient diagnosed with a cyanotic congenital heart disease of double-inlet left ventricle with TGA and unrepaired mitral stenosis, who underwent to a hybrid procedure of PAB and enlargement of the communication between the two atria.
ABSTRACT
En un esfuerzo por estandarizar el manejo perioperatorio y mejorar los resultados posoperatorios de los pacientes adultos sometidos a cirugía, el Ministerio de Sanidad, a través del Grupo Español de Rehabilitación Multimodal (GERM) y el Instituto Aragonés de Ciencias de la Salud, en colaboración con diversas sociedades científicas españolas, y sobre la base de la evidencia disponible, publicó en 2021 la guía Recuperación intensificada en cirugía del adulto (RICA). Dicho documento incluye 12 medidas perioperatorias relacionadas con la fluidoterapia y la monitorización hemodinámica. La administración de fluidos y la monitorización hemodinámica no son sencillas, pero están directamente relacionadas con los resultados de los pacientes. El Subcomité de Fluidoterapia y monitorización hemodinámica de la Sección de Hemostasia, Medicina transfusional y Fluidoterapia (SHTF) de la Sociedad Española de Anestesiología y Reanimación (SEDAR) ha analizado dichas recomendaciones, concluyendo que deberían ser revisadas, ya que no siguen la metodología adecuada. (AU)
In an effort to standardize perioperative management and improve postoperative outcomes of adult patients undergoing surgery, the Ministry of Health, through the Spanish Multimodal Rehabilitation Group (GERM), and the Aragonese Institute of Health Sciences, in collaboration with multiple Spanish scientific societies and based on the available evidence, published in 2021 the Spanish Intensified Adult Recovery (RICA) guideline. This document includes 12 perioperative measures related to fluid therapy and hemodynamic monitoring. Fluid administration and hemodynamic monitoring are not straightforward but are directly related to postoperative patient outcomes. The Fluid Therapy and Hemodynamic Monitoring Subcommittee of the Hemostasis, Transfusion Medicine and Fluid Therapy Section (SHTF) of the Spanish Society of Anesthesiology and Critical Care (SEDAR) has reviewed these recommendations and concluded that they should be revised as they do not follow an adequate methodology. (AU)
Subject(s)
Humans , Adult , Fluid Therapy , Hemodynamics , Perioperative Period/methods , Guidelines as Topic , Societies/standardsABSTRACT
In an effort to standardize perioperative management and improve postoperative outcomes of adult patients undergoing surgery, the Ministry of Health, through the Spanish Multimodal Rehabilitation Group (GERM), and the Aragonese Institute of Health Sciences, in collaboration with multiple Spanish scientific societies and based on the available evidence, published in 2021 the Spanish Intensified Adult Recovery (RICA) guideline. This document includes 12 perioperative measures related to fluid therapy and hemodynamic monitoring. Fluid administration and hemodynamic monitoring are not straightforward but are directly related to postoperative patient outcomes. The Fluid Therapy and Hemodynamic Monitoring Subcommittee of the Hemostasis, Transfusion Medicine and Fluid Therapy Section (SHTF) of the Spanish Society of Anesthesiology and Critical Care (SEDAR) has reviewed these recommendations and concluded that they should be revised as they do not follow an adequate methodology.
ABSTRACT
Antecedentes: La administración intraoperatoria de fluidos es una intervención ubicua en los pacientes quirúrgicos. Pero la administración inadecuada de fluidos puede llevar a malos resultados postoperatorios. La prueba de volumen (PV), dentro o fuera de la denominada fluidoterapia guiada por objetivos, permite probar el sistema cardiovascular y la necesidad de administración adicional de fluidos. Nuestro objetivo primario fue evaluar el modo en que el anestesiólogo realiza la PV en el quirófano en términos de tipo, volumen, variables utilizadas para impulsar la PV, y comparar la proporción de pacientes que recibieron administración adicional de fluidos basada en la respuesta a la PV. Métodos: Se trata de un subestudio planificado de un estudio observacional realizado en 131 centros en España, en pacientes sometidos a cirugía. Resultados: En el estudio se incluyeron y analizaron 396 pacientes. La cantidad media [rango intercuartílico] de fluidos administrados durante la PV fue de 250ml (200-400). La principal indicación de la PV fue el descenso de la presión arterial sistólica en 246 casos (62,2%). La segunda indicación fue el descenso de la presión arterial media (54,4%). Se utilizó el gasto cardiaco en 30 pacientes (7,58%), y la variación del volumen sistólico en 29 de entre 385 casos (7,32%). La respuesta a la PV inicial no tuvo impacto a la hora de prescribir administración adicional de fluidos. Conclusiones: La indicación y la evaluación actuales de la PV en los pacientes quirúrgicos son altamente variables. La predicción de la receptividad a los fluidos no se utiliza rutinariamente, evaluándose a menudo las variables no adecuadas para valorar la respuesta hemodinámica a la PV, pudiendo causar efectos perjudiciales.(AU)
Background: Intraoperative fluid administration is a ubiquitous intervention in surgical patients. But inadequate fluid administration may lead to poor postoperative outcomes. Fluid challenges (FCs), in or outside the so-called goal-directed fluid therapy, allows testing the cardiovascular system and the need for further fluid administration. Our primary aim was to evaluate how anesthesiologists conduct FCs in the operating room in terms of type, volume, variables used to trigger a FC and to compare the proportion of patients receiving further fluid administration based on the response to the FC. Methods: This was a planned substudy of an observational study conducted in 131 centers in Spain in patients undergoing surgery. Results: A total of 396 patients were enrolled and analyzed in the study. The median [interquartile range] amount of fluid given during a FC was 250mL (200-400). The main indication for FC was a decrease in systolic arterial pressure in 246 cases (62.2%). The second was a decrease in mean arterial pressure (54.4%). Cardiac output was used in 30 patients (7.58%), while stroke volume variation in 29 of 385 cases (7.32%). The response to the initial FC did not have an impact when prescribing further fluid administration. Conclusions: The current indication and evaluation of FC in surgical patients is highly variable. Prediction of fluid responsiveness is not routinely used, and inappropriate variables are frequently evaluated for assessing the hemodynamic response to FC, which may result in deleterious effects.(AU)
Subject(s)
Humans , Operating Rooms/methods , Noise , Intraoperative Complications , Intraoperative Period , Fluid Therapy/methods , Cohort Studies , Anesthesiology , General SurgeryABSTRACT
BACKGROUND: Intraoperative fluid administration is a ubiquitous intervention in surgical patients. But inadequate fluid administration may lead to poor postoperative outcomes. Fluid challenges (FCs), in or outside the so-called goal-directed fluid therapy, allows testing the cardiovascular system and the need for further fluid administration. Our primary aim was to evaluate how anesthesiologists conduct FCs in the operating room in terms of type, volume, variables used to trigger a FC and to compare the proportion of patients receiving further fluid administration based on the response to the FC. METHODS: This was a planned substudy of an observational study conducted in 131 centres in Spain in patients undergoing surgery. RESULTS: A total of 396 patients were enrolled and analysed in the study. The median [interquartile range] amount of fluid given during a FC was 250ml (200-400). The main indication for FC was a decrease in systolic arterial pressure in 246 cases (62.2%). The second was a decrease in mean arterial pressure (54.4%). Cardiac output was used in 30 patients (7.58%), while stroke volume variation in 29 of 385 cases (7.32%). The response to the initial FC did not have an impact when prescribing further fluid administration. CONCLUSIONS: The current indication and evaluation of FC in surgical patients is highly variable. Prediction of fluid responsiveness is not routinely used, and inappropriate variables are frequently evaluated for assessing the hemodynamic response to FC, which may result in deleterious effects.
Subject(s)
Fluid Therapy , Operating Rooms , Humans , Stroke Volume/physiology , Cardiac Output , HemodynamicsABSTRACT
BACKGROUND: Research in fluid therapy and perioperative hemodynamic monitoring is difficult and expensive. The objectives of this study were to summarize these topics and to prioritize these topics in order of research importance. METHODS: Electronic structured Delphi questionnaire over three rounds among 30 experts in fluid therapy and hemodynamic monitoring identified through the Fluid Therapy and Hemodynamic Monitoring Subcommittee of the Hemostasis, Transfusion Medicine and Fluid Therapy Section of the Spanish Society of Anesthesiology and Critical Care. RESULTS: 77 topics were identified and ranked in order of prioritization. Topics were categorized into themes of crystalloids, colloids, hemodynamic monitoring and others. 31 topics were ranked as essential research priority. To determine whether intraoperative hemodynamic optimization algorithms based on the invasive or noninvasive Hypotension Prediction Index versus other management strategies could decrease the incidence of postoperative complications. As well as whether the use of renal stress biomarkers together with a goal-directed fluid therapy protocol could reduce hospital stay and the incidence of acute kidney injury in adult patients undergoing non-cardiac surgery, reached the highest consensus. CONCLUSIONS: The Fluid Therapy and Hemodynamic Monitoring Subcommittee of the Hemostasis, Transfusion Medicine and Fluid Therapy Section of the Spanish Society of Anesthesiology and Critical Care will use these results to carry out the research.
Subject(s)
Anesthesiology , Hemodynamic Monitoring , Transfusion Medicine , Adult , Humans , Consensus , Delphi Technique , Fluid Therapy , Critical Care , HemostasisABSTRACT
Chromosomal rearrangements of the human KMT2A/MLL gene are associated with de novo as well as therapy-induced infant, pediatric, and adult acute leukemias. Here, we present the data obtained from 3401 acute leukemia patients that have been analyzed between 2003 and 2022. Genomic breakpoints within the KMT2A gene and the involved translocation partner genes (TPGs) and KMT2A-partial tandem duplications (PTDs) were determined. Including the published data from the literature, a total of 107 in-frame KMT2A gene fusions have been identified so far. Further 16 rearrangements were out-of-frame fusions, 18 patients had no partner gene fused to 5'-KMT2A, two patients had a 5'-KMT2A deletion, and one ETV6::RUNX1 patient had an KMT2A insertion at the breakpoint. The seven most frequent TPGs and PTDs account for more than 90% of all recombinations of the KMT2A, 37 occur recurrently and 63 were identified so far only once. This study provides a comprehensive analysis of the KMT2A recombinome in acute leukemia patients. Besides the scientific gain of information, genomic breakpoint sequences of these patients were used to monitor minimal residual disease (MRD). Thus, this work may be directly translated from the bench to the bedside of patients and meet the clinical needs to improve patient survival.
Subject(s)
Histone-Lysine N-Methyltransferase , Leukemia, Myeloid, Acute , Myeloid-Lymphoid Leukemia Protein , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Humans , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Leukemia, Myeloid, Acute/genetics , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Histone-Lysine N-Methyltransferase/genetics , Myeloid-Lymphoid Leukemia Protein/genetics , Gene FusionABSTRACT
JUSTIFICACIÓN: los agonistas del receptor del GLP-1 (aGLP-1) son un grupo de fármacos que reducen significativamente la hemoglobina glicosilada sin riesgo de hipoglucemias. Este grupo terapéutico tiene efectos multifactoriales más allá del control de la glucemia: disminuye el vaciamiento gástrico, aumenta la cardioprotección, etc. Una dispensación adecuada ayuda a mejorar la adherencia al tratamiento.OBJETIVOS: analizar el impacto del taller de intervención en la dispensación de los análogos de GLP-1 inyectables a través de unas preguntas de evaluación.MATERIAL Y MÉTODOS: se realizó el taller Intervención en la dispensación de análogos de GLP-1 inyectables en las diferentes sedes de SEFAC desde septiembre a diciembre del 2021. Para la elaboración del taller se siguió el siguiente procedimiento: Se seleccionaron 4 farmacéuticos expertos en el tema y miembros del grupo de diabetes de SEFAC para la creación del taller. La estructura a seguir fue: 10 minutos teoría, 20 minutos rol play y 10 minutos preguntas. Creación del taller y preguntas de evaluación, revisión por pares del material por comité científico de SEFAC y el comité médico del laboratorio patrocinador. Formación de los ponentes de las diferentes sedes de SEFAC a través de una sesión formativa de 2 horas vía zoom.Desarrollo del taller, realizando las MISMAS preguntas de evaluación antes y después del taller.RESULTADOS: 773 encuestas realizadas en las 15 sedes de SEFAC: 407 encuestas antes del taller y 366 después. El porcentaje de respuestas correctas fue de 50,0% antes y 64,3% después. (AU)
Subject(s)
Humans , Diabetes Mellitus , Hypoglycemia , Blood Glucose , Pharmacists , Treatment Adherence and ComplianceABSTRACT
Fusion oncogenes (FOs) are common in many cancer types and are powerful drivers of tumor development. Because their expression is exclusive to cancer cells and their elimination induces cell apoptosis in FO-driven cancers, FOs are attractive therapeutic targets. However, specifically targeting the resulting chimeric products is challenging. Based on CRISPR/Cas9 technology, here we devise a simple, efficient and non-patient-specific gene-editing strategy through targeting of two introns of the genes involved in the rearrangement, allowing for robust disruption of the FO specifically in cancer cells. As a proof-of-concept of its potential, we demonstrate the efficacy of intron-based targeting of transcription factors or tyrosine kinase FOs in reducing tumor burden/mortality in in vivo models. The FO targeting approach presented here might open new horizons for the selective elimination of cancer cells.
Subject(s)
CRISPR-Cas Systems/genetics , Neoplasms/genetics , Oncogene Fusion/genetics , Animals , Base Sequence , Cell Line, Tumor , Cell Proliferation/genetics , Doxorubicin/therapeutic use , Fusion Proteins, bcr-abl/genetics , Gene Deletion , Genetic Loci , Genomic Instability , HEK293 Cells , Humans , Introns/genetics , Mice, Nude , Neoplasms/drug therapy , Neoplasms/pathology , Oncogene Proteins, Fusion/genetics , RNA, Guide, Kinetoplastida/metabolism , Reproducibility of Results , Xenograft Model Antitumor AssaysABSTRACT
The original version of this Article contained an error in the spelling of the author Juan Carlos Rodriguez-Manzaneque, which was incorrectly given as J Carlos Rodríguez-Manzaneque. This has now been corrected in both the PDF and HTML versions of the Article.
ABSTRACT
Mixed-lineage leukemia (MLL)-rearranged (MLLr) infant B-cell acute lymphoblastic leukemia (iMLLr-B-ALL) has a dismal prognosis and is associated with a pro-B/mixed phenotype, therapy refractoriness and frequent central nervous system (CNS) disease/relapse. Neuron-glial antigen 2 (NG2) is specifically expressed in MLLr leukemias and is used in leukemia immunophenotyping because of its predictive value for MLLr acute leukemias. NG2 is involved in melanoma metastasis and brain development; however, its role in MLL-mediated leukemogenesis remains elusive. Here we evaluated whether NG2 distinguishes leukemia-initiating/propagating cells (L-ICs) and/or CNS-infiltrating cells (CNS-ICs) in iMLLr-B-ALL. Clinical data from the Interfant cohort of iMLLr-B-ALL demonstrated that high NG2 expression associates with lower event-free survival, higher number of circulating blasts and more frequent CNS disease/relapse. Serial xenotransplantation of primary MLL-AF4+ leukemias indicated that NG2 is a malleable marker that does not enrich for L-IC or CNS-IC in iMLLr-B-All. However, NG2 expression was highly upregulated in blasts infiltrating extramedullar hematopoietic sites and CNS, and specific blockage of NG2 resulted in almost complete loss of engraftment. Indeed, gene expression profiling of primary blasts and primografts revealed a migratory signature of NG2+ blasts. This study provides new insights on the biology of NG2 in iMLLr-B-ALL and suggests NG2 as a potential therapeutic target to reduce the risk of CNS disease/relapse and to provide safer CNS-directed therapies for iMLLr-B-ALL.
ABSTRACT
Chromosomal rearrangements of the human MLL/KMT2A gene are associated with infant, pediatric, adult and therapy-induced acute leukemias. Here we present the data obtained from 2345 acute leukemia patients. Genomic breakpoints within the MLL gene and the involved translocation partner genes (TPGs) were determined and 11 novel TPGs were identified. Thus, a total of 135 different MLL rearrangements have been identified so far, of which 94 TPGs are now characterized at the molecular level. In all, 35 out of these 94 TPGs occur recurrently, but only 9 specific gene fusions account for more than 90% of all illegitimate recombinations of the MLL gene. We observed an age-dependent breakpoint shift with breakpoints localizing within MLL intron 11 associated with acute lymphoblastic leukemia and younger patients, while breakpoints in MLL intron 9 predominate in AML or older patients. The molecular characterization of MLL breakpoints suggests different etiologies in the different age groups and allows the correlation of functional domains of the MLL gene with clinical outcome. This study provides a comprehensive analysis of the MLL recombinome in acute leukemia and demonstrates that the establishment of patient-specific chromosomal fusion sites allows the design of specific PCR primers for minimal residual disease analyses for all patients.
Subject(s)
Histone-Lysine N-Methyltransferase/genetics , Leukemia, Myeloid, Acute/genetics , Myeloid-Lymphoid Leukemia Protein/genetics , Adult , Child , Chromosome Aberrations , Chromosome Breakage , Female , Gene Rearrangement/genetics , Humans , Infant , Male , Oncogene Proteins, Fusion/genetics , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Translocation, Genetic/geneticsABSTRACT
BACKGROUND: To determine the knowledge and willingness of local police officers (PO) to perform cardiopulmonary resuscitation (CPR), as well as to explore the association between CPR training and these variables. METHODS: Cross-sectional study with a sample of 390 PO from Asturias (Spain). An anonymous questionnaire was used to measure nine basic aspects of CPR from the European Resuscitation Council and four indicators of attitude towards performing CPR in a real context. Information on CPR training and its periodicity was also collected, as well as basic socio-demographic and occupational variables. RESULTS: Lack of CPR training was seen in 19.7% of PO, and 36.4% had received such training more than two years ago. Almost one out of four PO had performed at least one CPR in a real situation (24.1%), of which 9.6% had not been trained. The least remembered aspects of CPR were depth (11%) and frequency of chest compressions (24.4%). Only 49.7% of the agents felt sufficiently prepared to perform a CPR. Knowledge and disposition were significantly associated with having received training with a periodicity of less than two years. CONCLUSIONS: Given that PO are frequently first responders in situations of out-of-hospital cardiorespiratory arrest, specific training in CPR should be mandatory and periodic, with at least one course every two years. It would be interesting to determine which didactic instrumentation is most efficient for disseminating these training courses among police officers. Key words. Police. Cardiopulmonary resuscitation. Out-of-hospital cardiac arrest; attitude. Emergencies.
Subject(s)
Attitude , Cardiopulmonary Resuscitation , Out-of-Hospital Cardiac Arrest/therapy , Police , Adolescent , Adult , Cardiopulmonary Resuscitation/education , Cross-Sectional Studies , Female , Humans , Male , Middle Aged , Spain , Young AdultABSTRACT
Fundamento: Determinar los conocimientos de los agentes de la policía local (PL) sobre la reanimación cardiopulmonar (RCP) y su disposición para realizarla, así como explorar la asociación entre la formación en RCP y estas variables. Sujetos y método: Estudio transversal sobre una muestra de 390 agentes de la PL de Asturias (España). Se utilizó un cuestionario anónimo que midió nueve aspectos básicos sobre la RCP del Consejo Europeo de Reanimación y cuatro indicadores de la disposición para realizarla en situaciones reales. También se recogió información sobre la formación en RCP y su periodicidad, así como variables sociodemográficas y laborales básicas. Resultados: El 19,7% de los PL no había recibido formación en RCP y el 36,4% la había recibido hacía más de dos. El 24,1% habían realizado al menos una RCP en situación real, de los cuales el 9,6% no estaba formado. Los aspectos de la RCP menos recordados fueron la profundidad (11%) y la frecuencia de las compresiones (24,4%). El 49,7% de los agentes se siente con suficiente preparación para realizar una RCP. Los conocimientos y la disposición se asociaron significativamente con haber realizado cursos de formación con una periodicidad menor de dos años. Conclusiones: Dado que los agentes de PL deben intervenir con frecuencia en situaciones de parada cardiorrespiratoria como primeros intervinientes, la formación específica en RCP de los agentes de PL debería ser obligatoria y periódica, con al menos un curso cada dos años. Sería interesante determinar qué instrumentación didáctica es más eficiente para difundir estos cursos entre los policías (AU)
Background: To determine the knowledge and willingness of local police officers (PO) to perform cardiopulmonary resuscitation (CPR), as well as to explore the association between CPR training and these variables. Methods: Cross-sectional study with a sample of 390 PO from Asturias (Spain). An anonymous questionnaire was used to measure nine basic aspects of CPR from the European Resuscitation Council and four indicators of attitude towards performing CPR in a real context. Information on CPR training and its periodicity was also collected, as well as basic socio-demographic and occupational variables. Results: Lack of CPR training was seen in 19.7% of PO, and 36.4% had received such training more than two years ago. Almost one out of four PO had performed at least one CPR in a real situation (24.1%), of which 9.6% had not been trained. The least remembered aspects of CPR were depth (11%) and frequency of chest compressions (24.4%). Only 49.7% of the agents felt sufficiently prepared to perform a CPR. Knowledge and disposition were significantly associated with having received training with a periodicity of less than two years. Conclusions: Given that PO are frequently first responders in situations of out-of-hospital cardiorespiratory arrest, specific training in CPR should be mandatory and periodic, with at least one course every two years. It would be interesting to determine which didactic instrumentation is most efficient for disseminating these training courses among police officers (AU)
Subject(s)
Humans , Male , Female , Young Adult , Adult , Middle Aged , Vital Signs/physiology , Life Support Systems/standards , Police/statistics & numerical data , Health Knowledge, Attitudes, Practice , Cardiopulmonary Resuscitation/instrumentation , Ambulatory Care/standards , Emergency Medical Services/organization & administration , First Aid/instrumentation , Health Systems/organization & administration , Health Systems/standardsABSTRACT
B cells have been shown to be refractory to reprogramming and B-cell-derived induced pluripotent stem cells (iPSC) have only been generated from murine B cells engineered to carry doxycycline-inducible Oct4, Sox2, Klf4 and Myc (OSKM) cassette in every tissue and from EBV/SV40LT-immortalized lymphoblastoid cell lines. Here, we show for the first time that freshly isolated non-cultured human cord blood (CB)- and peripheral blood (PB)-derived CD19+CD20+ B cells can be reprogrammed to iPSCs carrying complete VDJH immunoglobulin (Ig) gene monoclonal rearrangements using non-integrative tetracistronic, but not monocistronic, OSKM-expressing Sendai Virus. Co-expression of C/EBPα with OSKM facilitates iPSC generation from both CB- and PB-derived B cells. We also demonstrate that myeloid cells are much easier to reprogram than B and T lymphocytes. Differentiation potential back into the cell type of their origin of B-cell-, T-cell-, myeloid- and fibroblast-iPSCs is not skewed, suggesting that their differentiation does not seem influenced by 'epigenetic memory'. Our data reflect the actual cell-autonomous reprogramming capacity of human primary B cells because biased reprogramming was avoided by using freshly isolated primary cells, not exposed to cytokine cocktails favoring proliferation, differentiation or survival. The ability to reprogram CB/PB-derived primary human B cells offers an unprecedented opportunity for studying developmental B lymphopoiesis and modeling B-cell malignancies.
Subject(s)
B-Lymphocytes/metabolism , CCAAT-Enhancer-Binding Proteins/genetics , Cellular Reprogramming/genetics , Fetal Blood/metabolism , Induced Pluripotent Stem Cells/metabolism , Leukocytes, Mononuclear/metabolism , B-Lymphocytes/cytology , B-Lymphocytes/immunology , Base Sequence , CCAAT-Enhancer-Binding Proteins/immunology , Cell Differentiation , Cell Separation , Cellular Reprogramming/immunology , Fetal Blood/cytology , Fetal Blood/immunology , Gene Expression , Genetic Vectors , Humans , Induced Pluripotent Stem Cells/cytology , Induced Pluripotent Stem Cells/immunology , Kruppel-Like Factor 4 , Kruppel-Like Transcription Factors/genetics , Kruppel-Like Transcription Factors/immunology , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/immunology , Molecular Sequence Data , Myeloid Cells/cytology , Myeloid Cells/immunology , Myeloid Cells/metabolism , Octamer Transcription Factor-3/genetics , Octamer Transcription Factor-3/immunology , Primary Cell Culture , Proto-Oncogene Proteins c-myc/genetics , Proto-Oncogene Proteins c-myc/immunology , SOXB1 Transcription Factors/genetics , SOXB1 Transcription Factors/immunology , Sendai virus/genetics , V(D)J Recombination/immunologyABSTRACT
Notch signaling is essential for definitive hematopoiesis, but its role in human embryonic hematopoiesis is largely unknown. We show that in hESCs the expression of the Notch ligand DLL4 is induced during hematopoietic differentiation. We found that DLL4 is only expressed in a sub-population of bipotent hematoendothelial progenitors (HEPs) and segregates their hematopoietic versus endothelial potential. We demonstrate at the clonal level and through transcriptome analyses that DLL4(high) HEPs are enriched in endothelial potential, whereas DLL4(low/-) HEPs are committed to the hematopoietic lineage, albeit both populations still contain bipotent cells. Moreover, DLL4 stimulation enhances hematopoietic differentiation of HEPs and increases the amount of clonogenic hematopoietic progenitors. Confocal microscopy analysis of whole differentiating embryoid bodies revealed that DLL4(high) HEPs are located close to DLL4(low/-) HEPs, and at the base of clusters of CD45+ cells, resembling intra-aortic hematopoietic clusters found in mouse embryos. We propose a model for human embryonic hematopoiesis in which DLL4(low/-) cells within hemogenic endothelium receive Notch-activating signals from DLL4(high) cells, resulting in an endothelial-to-hematopoietic transition and their differentiation into CD45+ hematopoietic cells.
Subject(s)
Biomarkers/metabolism , Cell Differentiation , Embryonic Stem Cells/cytology , Endothelium/cytology , Hematopoiesis/physiology , Hematopoietic Stem Cells/cytology , Intracellular Signaling Peptides and Proteins/metabolism , Membrane Proteins/metabolism , Animals , Cell Proliferation , Cells, Cultured , Embryoid Bodies , Embryonic Stem Cells/metabolism , Endothelium/metabolism , Female , Flow Cytometry , Gene Expression Profiling , Hematopoietic Stem Cells/metabolism , Humans , Immunoenzyme Techniques , Intracellular Signaling Peptides and Proteins/genetics , Membrane Proteins/genetics , Mice , Mice, Inbred NOD , Mice, SCID , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Natural killer cells (NK) are important effectors of anti-tumor immunity, activated either by the downregulation of HLA-I molecules on tumor cells and/or the interaction of NK-activating receptors with ligands that are overexpressed on target cells upon tumor transformation (including NKG2D and NKP30). NK kill target cells by the vesicular delivery of cytolytic molecules such as Granzyme-B and Granulysin activating different cell death pathways, which can be Caspase-3 dependent or Caspase-3 independent. Multiple myeloma (MM) remains an incurable neoplastic plasma-cell disorder. However, we previously reported the encouraging observation that cord blood-derived NK (CB-NK), a new source of NK, showed anti-tumor activity in an in vivo murine model of MM and confirmed a correlation between high levels of NKG2D expression by MM cells and increased efficacy of CB-NK in reducing tumor burden. We aimed to characterize the mechanism of CB-NK-mediated cytotoxicity against MM cells. We show a Caspase-3- and Granzyme-B-independent cell death, and we reveal a mechanism of transmissible cell death between cells, which involves lipid-protein vesicle transfer from CB-NK to MM cells. These vesicles are secondarily transferred from recipient MM cells to neighboring MM cells amplifying the initial CB-NK cytotoxicity achieved. This indirect cytotoxicity involves the transfer of NKG2D and NKP30 and leads to lysosomal cell death and decreased levels of reactive oxygen species in MM cells. These findings suggest a novel and unique mechanism of CB-NK cytotoxicity against MM cells and highlight the importance of lipids and lipid transfer in this process. Further, these data provide a rationale for the development of CB-NK-based cellular therapies in the treatment of MM.
Subject(s)
Immunity, Cellular , Killer Cells, Natural/immunology , Multiple Myeloma/immunology , Secretory Vesicles/immunology , Caspase 3/immunology , Female , Fetal Blood , Granzymes/immunology , Humans , K562 Cells , Killer Cells, Natural/pathology , Male , Multiple Myeloma/pathology , NK Cell Lectin-Like Receptor Subfamily K/immunology , Natural Cytotoxicity Triggering Receptor 3/immunology , Reactive Oxygen Species/immunologyABSTRACT
La neumatosis gástrica es un hallazgo radiológico infrecuente que puede deberse a causas de diferente pronóstico. Es preciso un diagnóstico diferencial precoz que permita establecer las pautas terapéuticas adecuadas, dadas las altas tasas de mortalidad asociadas
Gastric pneumatosis is an uncommon radiological finding due to causes of different prognosis. It is necessary an early differential diagnosis that allows the establishment of an appropriated treatment, due to high mortality rates associated
