Relevance of Thymic Stromal Lymphopoietin on the Pathogenesis of Glioblastoma: Role of the Neutrophil.

Glioblastoma multiforme (GBM) is the most predominant and malignant primary brain tumor in adults. Thymic stromal lymphopoietin (TSLP), a cytokine primarily generated by activated epithelial cells, has recently garnered attention in cancer research. This study was aimed to elucidate the significance of TSLP in GBM cells and its interplay with the immune system, particularly focused on granulocyte neutrophils. Our results demonstrate that the tumor produces TSLP when stimulated with epidermal growth factor (EGF) in both the U251 cell line and the GBM biopsy (GBM-b). The relevance of the TSLP function was evaluated using a 3D spheroid model. Spheroids exhibited increased diameter, volume, and proliferation. In addition, TSLP promoted the generation of satellites surrounding the main spheroids and inhibited apoptosis in U251 treated with temozolomide (TMZ). Additionally, the co-culture of polymorphonuclear (PMN) cells from healthy donors with the U251 cell line in the presence of TSLP showed a reduction in apoptosis and an increase in IL-8 production. TSLP directly inhibited apoptosis in PMN from GBM patients (PMN-p). Interestingly, the vascular endothelial growth factor (VEGF) production was elevated in PMN-p compared with PMN from healthy donors. Under these conditions, TSLP also increased VEGF production, in PMN from healthy donors. Moreover, TSLP upregulated programed death-ligand 1 (PDL-1) expression in PMN cultured with U251. On the other hand, according to our results, the analysis of RNA-seq datasets from Illumina HiSeq 2000 sequencing platform performed with TIMER2.0 webserver demonstrated that the combination of TSLP with neutrophils decreases the survival of the patient. In conclusion, our results position TSLP as a possible new growth factor in GBM and indicate its modulation of the tumor microenvironment, particularly through its interaction with PMN.

A spatial vascular transcriptomic, proteomic, and phosphoproteomic atlas unveils an angiocrine Tie-Wnt signaling axis in the liver.

Single-cell transcriptomics (scRNA-seq) has revolutionized the understanding of the spatial architecture of tissue structure and function. Advancing the "transcript-centric" view of scRNA-seq analyses is presently restricted by the limited resolution of proteomics and genome-wide techniques to analyze post-translational modifications. Here, by combining spatial cell sorting with transcriptomics and quantitative proteomics/phosphoproteomics, we established the spatially resolved proteome landscape of the liver endothelium, yielding deep mechanistic insight into zonated vascular signaling mechanisms. Phosphorylation of receptor tyrosine kinases was detected preferentially in the central vein area, resulting in an atypical enrichment of tyrosine phosphorylation. Prototypic biological validation identified Tie receptor signaling as a selective and specific regulator of vascular Wnt activity orchestrating angiocrine signaling, thereby controlling hepatocyte function during liver regeneration. Taken together, the study has yielded fundamental insight into the spatial organization of liver endothelial cell signaling. Spatial sorting may be employed as a universally adaptable strategy for multiomic analyses of scRNA-seq-defined cellular (sub)-populations.

The p.(Cys150Tyr) variant in CSRP3 is associated with late-onset hypertrophic cardiomyopathy in heterozygous individuals.

INTRODUCTION AND OBJECTIVES: Up to 50% of patients with hypertrophic cardiomyopathy (HCM) show no disease-causing variants in genetic studies. Mutations in CSRP3 have been associated with HCM, but evidence supporting pathogenicity is inconclusive. In this study, we describe an HCM cohort with a missense variant in CSRP3 (p.Cys150Tyr) with supporting evidence for pathogenicity and a description of the associated phenotype. METHODS: CSRP3 was sequenced in 6456 index cases with a diagnosis of HCM and in 5012 probands with other cardiomyopathies. In addition, 3372 index cases with hereditary cardiovascular disorders other than cardiomyopathies (mainly channelopathies and aortopathies) were used as controls. RESULTS: The p.(Cys150Tyr) variant was identified in 11 unrelated individuals of the 6456 HCM probands, and it was not identified in patients with other cardiomyopathies (p < 0.0001) or in our control population (p < 0.0001). Ten of the index cases were heterozygous and one was homozygous. Homozygous had a more severe phenotype. Family screening identified 17 other carriers. Wild-type individuals showed no signs of disease. The mean age at diagnosis of affected individuals was 55 ±â€¯13 years, and the mean left ventricular wall thickness was 18 ±â€¯3 mm. The variant showed highly age-dependent penetrance. After a mean follow-up of 11 (±8) years, no adverse events were reported in any of the HCM patients. CONCLUSIONS: The p.(Cys150Tyr) variant in CSRP3 causes late-onset and low risk form of hypertrophic cardiomyopathy in heterozygous carriers.

Alteration of platelet GPVI signaling in ST-elevation myocardial infarction patients demonstrated by a combination of proteomic, biochemical, and functional approaches.

The platelet-specific collagen receptor glycoprotein VI (GPVI) is critical for the formation of arterial thrombosis in vivo. We analyzed GPVI-activated platelets from ST-elevation myocardial infarction (STEMI) patients and matched stable coronary artery disease (SCAD) controls in order to provide novel clues on the degree of involvement of GPVI signaling in the acute event. Firstly, platelets were isolated from systemic venous blood and activated with the GPVI specific agonist CRP (collagen-related peptide). STEMI and SCAD samples were compared by a phosphoproteomics approach. Validations were by immunoblotting in systemic and intracoronary blood from independent cohorts of patients. Twenty-six differentially regulated proteins were identified when comparing CRP-activated systemic platelets from STEMI and SCAD patients, 4 of which were selected for validation studies: PLCÉ£2, G6f, SLP-76, and Dok-2. Immunoblot analyses showed these four proteins had higher tyrosine phosphorylation levels in response to CRP in platelets from STEMI patients, being these levels more pronounced at the culprit site of coronary artery occlusion. Moreover, platelet aggregation studies showed a higher response to GPVI agonists in STEMI patients compared to SCAD controls. In conclusion, we show an altered activation state of GPVI signaling in STEMI patients, confirming this receptor as a promising anti-thrombotic target for myocardial infarction.

Calidad de vida relacionada con la salud en adolescentes con sobrepeso y obesidad: casos y controles / Quality of life related to health in overweight and obese adolescents: cases and controls

ResumenObjetivo: Determinar la asociación entre la calidad de vida relacionada con la salud, con el sobrepeso y la obesidad en adolescentesMaterial y Método:estudio de casos y controles, 256 adolescentes entre 15 y 19 años; se les aplicó un instrumento validado, cuestionario de salud SF-12 y cuestionario Kiddo-Kindl para jóvenes.Resultados:Promedio de edad de los adolescentes fue de 17 años, promedio de IMC 24,27 kg/m2 +/-3,82 kg/m2. Se encontró que la dimensión del SF-12 con menor promedio de puntuación fue salud general (66,80±20,076). Las dimensiones mejor percibidas fueron: rol físico (89,45±26,311) y función física (87,21 ±23,638). Los mejores resultados en la valoración de la calidad de vida, estuvieron en las dimensiones familia (4,08±0,86) y amigos (4,08±0,80), lo relacionado con la participación obtuvo significancia estadística (p < 0,05), esta se comporta como factor de riesgo para la presencia del sobrepeso y la obesidad, así como el estar orgulloso de la apariencia física, (OR = 4,31, IC = 1,10-19,77) (p < 0,05).Conclusión:Las dimensiones de calidad de vida, que mejor autoperciben los adolescentes fueron: rol físico, y dimensión corporal. La dimensión rol emocional fue estadísticamente significativa para los casos, y para controles fue la función social. En la edad, para el grupo de casos la dimensión que tuvo diferencias estadísticamente significativas fue la corporal, y en los controles la dimensión vitalidad. La percepción de la calidad de vida antes de presentar sobrepeso fue buena, contrario a lo manifestado después de presentar sobrepeso y obesidad en los participantes.

AbstractObjective: To determine the association between quality of life related to overweight and obesity in adolescents.Material and Methods: A case-control study of 256 adolescents between 15 and 19 years were administered a validated instrument, the SF -12 health questionnaire and Kiddo - Kindl questionnaire for young people.Results: The average age of the adolescents was 17, the average BMI was 24.27 kg/m2 +/-3.82 kg/m2. The dimension of the SF-12 with the lowest average score was general health (66.80±20.076). The best perceived dimensions were: physical role (89.45 ± 26.311) and physical functioning (87.21 ± 23.638). The best results in the assessment of quality of life were in the dimension's family (4.08 ± 0.86) and friends (4.08 ± 0.80); matters related to participation obtained statistical significance (p < 0.05), which behaves as a risk factor for the presence of overweight and obesity, as well as being proud of physical appearance (OR = 4.31, IC = 1.10 to 19.77) (p < 0.05).Conclusion: The dimensions of quality of life, which are best perceived by teenagers, were physical role, and bodily dimension. The emotional role dimension was statistically significant in the test cases, and in the control cases, social function. Regarding age, body was the statistically significant dimension for the test group, and vitality for the control group. The perception of quality of life before being overweight was good, in contrast to the perception of quality of life after being overweight and obese.

2D-DIGE-based proteomic analysis of intracoronary versus peripheral arterial blood platelets from acute myocardial infarction patients: Upregulation of platelet activation biomarkers at the culprit site.

PURPOSE: Platelets play a fundamental role in the atherothrombotic events that lead to an acute myocardial infarction. In the present study we compared the proteome of intracoronary and peripheral arterial platelets from ST-elevation myocardial infarction (STEMI) patients in the search for potential platelet biomarkers/drug targets related to what is happening at the culprit site. EXPERIMENTAL DESIGN: Ten STEMI patients were recruited and blood collected from the occluded coronary artery, at the culprit site, in the moment of reperfusion. Systemic blood obtained from the radial artery of the same patients was used as control. Proteome analysis was based on high-resolution 2D-DIGE and mass spectrometry. Validations were by western blotting in a group of 11 patients. RESULTS: Sixteen differentially regulated protein features were identified, corresponding to 15 ORFs, mostly related to cytoskeletal and signaling proteins. We demonstrate the up-regulation of integrin αIIb (ITA2B), the adapter Src kinase-associated phosphoprotein-2 (SKAP2), and thrombospondin-1 isoforms in intracoronary platelets. CONCLUSION AND CLINICAL RELEVANCE: This study constitutes the first analyzing in detail the proteome of arterial intracoronary platelets from STEMI patients. We show variations in the platelet proteome when comparing intracoronary and peripheral platelets. Observed differences might be related to platelet activation events at the culprit site.

A 2D-DIGE-based proteomic analysis reveals differences in the platelet releasate composition when comparing thrombin and collagen stimulations.

Upon stimulation, platelets release a high number of proteins (the releasate). There are clear indications that these proteins are involved in the pathogenesis of several diseases, such as atherosclerosis. In the present study we compared the platelet releasate following platelet activation with two major endogenous agonists: thrombin and collagen. Proteome analysis was based on 2D-DIGE and LC-MS/MS. Firstly, we showed the primary role of thrombin and collagen receptors in platelet secretion by these agonists; moreover, we demonstrated that GPVI is the primary responsible for collagen-induced platelet activation/aggregation. Proteomic analysis allowed the detection of 122 protein spots differentially regulated between both conditions. After excluding fibrinogen spots, down-regulated in the releasate of thrombin-activated platelets, 84 differences remained. From those, we successfully identified 42, corresponding to 37 open-reading frames. Many of the differences identified correspond to post-translational modifications, primarily, proteolysis induced by thrombin. Among others, we show vitamin K-dependent protein S, an anticoagulant plasma protein, is up-regulated in thrombin samples. Our results could have pathological implications given that platelets might be playing a differential role in various diseases and biological processes through the secretion of different subsets of granule proteins and microvesicles following a predominant activation of certain receptors.

Identification of a circulating microvesicle protein network involved in ST-elevation myocardial infarction.

Membrane microvesicles (MVs) are released from activated cells, most notably platelets, into the circulation. They represent an important mode of intercellular communication, and their number is increased in patients with acute coronary syndromes. We present here a differential proteomic analysis of plasma MVs from ST-elevation myocardial infarction (STEMI) patients and stable coronary artery disease (SCAD) controls. The objective was the identification of MVs biomarkers/drug targets that could be relevant for the pathogenesis of the acute event. Proteome analysis was based on 2D-DIGE, and mass spectrometry. Validations were by western blotting in an independent cohort of patients and healthy individuals. A systems biology approach was used to predict protein-protein interactions and their relation with disease. Following gel image analysis, we detected 117 protein features that varied between STEMI and SCAD groups (fold change cut-off ≥2; p<0.01). From those, 102 were successfully identified, corresponding to 25 open-reading frames (ORFs). Most of the proteins identified are involved in inflammatory response and cardiovascular disease, with 11 ORFs related to infarction. Among others, we report an up-regulation of α2-macroglobulin isoforms, fibrinogen, and viperin in MVs from STEMI patients. Interestingly, several of the proteins identified are involved in thrombogenesis (e.g. α2-macroglobulin, and fibrinogen). In conclusion, we provide a unique panel of proteins that vary between plasma MVs from STEMI and SCAD patients and that might constitute a promising source of biomarkers/drug targets for myocardial infarction.

A detailed proteomic analysis of rhodocytin-activated platelets reveals novel clues on the CLEC-2 signalosome: implications for CLEC-2 signaling regulation.

C-type lectin-like receptor 2 (CLEC-2) is an essential platelet-activating receptor in hemostasis and thrombosis that is activated by the snake venom rhodocytin. We present here a differential proteomic analysis of basal and rhodocytin-activated platelets with the aim of providing novel clues on CLEC-2 signaling regulation. Proteome analysis was based on 2D-DIGE, phosphotyrosine immunoprecipitations followed by 1D SDS-PAGE and mass spectrometry. Protein-protein interactions were studied by coimmunoprecipitations and a systems biology approach. Overall, we identified 132 proteins differentially regulated after CLEC-2 platelet activation, including most of the major players reported so far in the signaling cascade. In addition, we identified various proteins not previously known to participate in CLEC-2 signaling, such as the adapters Dok-2 and ADAP, tyrosine kinase Fer, and tyrosine phosphatase SHIP-1. We also report an increased association between Dok-2 and SHIP-1 in rhodocytin-stimulated platelets, which might negatively regulate CLEC-2 signaling. Moreover, we also present a comparative analysis of proteomic data for CLEC-2 and glycoprotein VI signaling. We think that our data provide thrombosis-relevant information on CLEC-2 signaling regulation, contributing to a better understanding of this important signaling cascade.

Conocimiento y aplicación del derecho internacional humanitario y la misión médica en los servicios de urgencias del oriente antioqueño / Knowledge and application of the International Human Right and the Medical Mission in the emergency services in the East Region of Antioquia

En octubre de 2002 se llevó a cabo un estudio descriptivo de corte con el propósito de investigar sobre el conocimiento y la aplicación del Derecho Internacional Humanitario (DIH) y la Misión Médica (MM), en los servicios de urgencias localizados en municipios del oriente del departamento de Antioquia, región que sufre desde hace varios años las consecuencias del conflicto armado colombiano y que refleja la problemática general del país. El estudio se hizo encuestando a los directores de esos servicios de urgencias para averiguar qué formación habían tenido sobre DIH y MM y qué pautas había en las instituciones que representaban para la aplicación de estas normas. Los directores encuestados manifestaron tener muy poca información acerca del DIH y la MM. Durante su formación académica de pregrado y posgrado no recibieron educación formal al respecto. Estuvieron de acuerdo en la importancia del tema, la necesidad de capacitarse y la posibilidad de aplicarlo. Por otra parte, en las instituciones de salud que cuentan con servicios de urgencias en el oriente antioqueño se carece de protocolos o manuales sobre el DIH y la MM así como de actividades de formación continua que permitan al personal sanitario actuar a la luz de estas normas; además, el conocimiento previo de estos aspectos no es requisito para laborar en las instituciones de salud que cuentan con servicios de urgencias en la región.

In october ctober 2002 a descriptive study was made to find out the grade of knowledge in the Emergency Services of the East region of Antioquia of the International Human Right, (IHR) and the Medical Mission (MM). This region has been suffering since many years the consequences of the war that show what has been happening in the entire country. A questionnaire was filled by the directors of the hospitals in this region about both items. They agree on the importance of the items but said they have little knowledge about them and they have received few instructions regarding those items. In their hospitals there are neither guides nor education programs for the personnel in this regard. To be posted in a hospital, a person needs not show proficiency in IHR or MM

Estudio cefalométrico longitudinal de la variabilidad en el crecimiento craneofacial en 55 pacientes de la comunidad de Damasco Antioquia / Longitudinal cephalometirc study of the variability of craniofacial growth in 55 patients of the community of Damasco Colombia

Este es un estudio cefalométrico longitudinal a 10 años en 55 individuos, niños de la comunidad de Damasco, corregimiento de Santa Bárbara Antioquia. En Colombia no se han realizado estudios de tipo longitudinal; los estudios reportados hasta hoy son de tipo transversal como el de Zagarra, y Villegas en Bogotá; Cárdenas en Heliconia Antioquia, Y Palacino en Medellín. Los objetivos de este estudio fueron observar y evaluar el comportamiento del crecimiento craneofacial longitudinal en niños entre los 6 y los 13 años de edad de la comunidad de Damasco, comparar los resultados con los reportados por otros estudios y establecer diferencias. La recolección de datos se hizo por medio de radiografía cefálica lateral tomadas a cada individuo con un intervalo de 2 años. Para el análisis de las radiografías se tuvieron en cuenta 10 mediciones en estructuras óseas, nueve lineales y una angular. Al analizar los resultados se observó que existen diferencias entre individuos de la comunidad de Damasco y los pertenecientes a los demás estudios encontrándose para los de Damasco valores menores en la mayoría de las dimensiones. Además se observaron diferencias entre los individuos de ambos sexos para todos los estudios, siendo mayores los valores para los hombres que para las mujeres...