Association Between Preexisting Versus Newly Identified Atrial Fibrillation and Outcomes of Patients With Acute Pulmonary Embolism

Background Atrial fibrillation (AF) may exist before or occur early in the course of pulmonary embolism (PE). We determined the PE outcomes based on the presence and timing of AF. Methods and Results Using the data from a multicenter PE registry, we identified 3 groups: (1) those with preexisting AF, (2) patients with new AF within 2 days from acute PE (incident AF), and (3) patients without AF. We assessed the 90-day and 1-year risk of mortality and stroke in patients with AF, compared with those without AF (reference group). Among 16 497 patients with PE, 792 had preexisting AF. These patients had increased odds of 90-day all-cause (odds ratio [OR], 2.81; 95% CI, 2.33-3.38) and PE-related mortality (OR, 2.38; 95% CI, 1.37-4.14) and increased 1-year hazard for ischemic stroke (hazard ratio, 5.48; 95% CI, 3.10-9.69) compared with those without AF. After multivariable adjustment, preexisting AF was associated with significantly increased odds of all-cause mortality (OR, 1.91; 95% CI, 1.57-2.32) but not PE-related mortality (OR, 1.50; 95% CI, 0.85-2.66). Among 16 497 patients with PE, 445 developed new incident AF within 2 days of acute PE. Incident AF was associated with increased odds of 90-day all-cause (OR, 2.28; 95% CI, 1.75-2.97) and PE-related (OR, 3.64; 95% CI, 2.01-6.59) mortality but not stroke. Findings were similar in multivariable analyses. Conclusions In patients with acute symptomatic PE, both preexisting AF and incident AF predict adverse clinical outcomes. The type of adverse outcomes may differ depending on the timing of AF onset.info:eu-repo/semantics/publishedVersio

Multiplex Analysis of CircRNAs from Plasma Extracellular Vesicle-Enriched Samples for the Detection of Early-Stage Non-Small Cell Lung Cancer

Background: The analysis of liquid biopsies brings new opportunities in the precision oncology field. Under this context, extracellular vesicle circular RNAs (EV-circRNAs) have gained interest as biomarkers for lung cancer (LC) detection. However, standardized and robust protocols need to be developed to boost their potential in the clinical setting. Although nCounter has been used for the analysis of other liquid biopsy substrates and biomarkers, it has never been employed for EV-circRNA analysis of LC patients. Methods: EVs were isolated from early-stage LC patients (n = 36) and controls (n = 30). Different volumes of plasma, together with different number of pre-amplification cycles, were tested to reach the best nCounter outcome. Differential expression analysis of circRNAs was performed, along with the testing of different machine learning (ML) methods for the development of a prognostic signature for LC. Results: A combination of 500 μL of plasma input with 10 cycles of pre-amplification was selected for the rest of the study. Eight circRNAs were found upregulated in LC. Further ML analysis selected a 10-circRNA signature able to discriminate LC from controls with AUC ROC of 0.86. Conclusions: This study validates the use of the nCounter platform for multiplexed EV-circRNA expression studies in LC patient samples, allowing the development of prognostic signatures

KRAS G12C-mutant driven non-small cell lung cancer (NSCLC)

KRAS G12C mutations in non-small cell lung cancer (NSCLC) partially respond to KRAS G12C covalent inhibitors. However, early adaptive resistance occurs due to rewiring of signaling pathways, activating receptor tyrosine kinases, primarily EGFR, but also MET and ligands. Evidence indicates that treatment with KRAS G12C inhibitors (sotorasib) triggers the MRAS:SHOC2:PP1C trimeric complex. Activation of MRAS occurs from alterations in the Scribble and Hippo-dependent pathways, leading to YAP activation. Other mechanisms that involve STAT3 signaling are intertwined with the activation of MRAS. The high-resolution MRAS:SHOC2:PP1C crystallization structure allows in silico analysis for drug development. Activation of MRAS:SHOC2:PP1C is primarily Scribble-driven and downregulated by HUWE1. The reactivation of the MRAS complex is carried out by valosin containing protein (VCP). Exploring these pathways as therapeutic targets and their impact on different chemotherapeutic agents (carboplatin, paclitaxel) is crucial. Comutations in STK11/LKB1 often co-occur with KRAS G12C, jeopardizing the effect of immune checkpoint (anti-PD1/PDL1) inhibitors

RNA-Based Multiplexing Assay for Routine Testing of Fusion and Splicing Variants in Cytological Samples of NSCLC Patients

The detection of ALK receptor tyrosine kinase (ALK), ROS proto-oncogen1, receptor tyrosine kinase (ROS1), ret proto-oncogen (RET), and MET proto-oncogen exon 14 skipping (METΔex14) allows for the selection of specific kinase inhibitor treatment in patients with non-small cell lung cancer (NSCLC). Multiplex technologies are recommended in this setting. We used nCounter, a multiplexed technology based on RNA hybridization, to detect ALK, ROS1, RET, and METΔex14 in RNA purified from cytological specimens (n = 16) and biopsies (n = 132). Twelve of the 16 cytological samples (75.0%) were evaluable by nCounter compared to 120 out of 132 (90.9%) biopsies. The geometrical mean (geomean) of the housekeeping genes of the nCounter panel, but not the total amount of RNA purified, was significantly higher in biopsies vs. cytological samples. Among cytological samples, we detected ALK (n = 3), METΔex14 (n = 1) and very high MET expression (n = 1) positive cases. The patient with METΔex14 had a partial response to tepotinib, one of the patients with ALK fusions was treated with crizotinib with a complete response. Cell blocks and cytological extensions can be successfully used for the detection of fusions and splicing variants using RNA-based methods such as nCounter

Multiplex Analysis of CircRNAs from Plasma Extracellular Vesicle-Enriched Samples for the Detection of Early-Stage Non-Small Cell Lung Cancer

Background: The analysis of liquid biopsies brings new opportunities in the precision oncology field. Under this context, extracellular vesicle circular RNAs (EV-circRNAs) have gained interest as biomarkers for lung cancer (LC) detection. However, standardized and robust protocols need to be developed to boost their potential in the clinical setting. Although nCounter has been used for the analysis of other liquid biopsy substrates and biomarkers, it has never been employed for EV-circRNA analysis of LC patients. Methods: EVs were isolated from early-stage LC patients (n = 36) and controls (n = 30). Different volumes of plasma, together with different number of pre-amplification cycles, were tested to reach the best nCounter outcome. Differential expression analysis of circRNAs was performed, along with the testing of different machine learning (ML) methods for the development of a prognostic signature for LC. Results: A combination of 500 μL of plasma input with 10 cycles of pre-amplification was selected for the rest of the study. Eight circRNAs were found upregulated in LC. Further ML analysis selected a 10-circRNA signature able to discriminate LC from controls with AUC ROC of 0.86. Conclusions: This study validates the use of the nCounter platform for multiplexed EV-circRNA expression studies in LC patient samples, allowing the development of prognostic signatures

Multiplex Detection of Clinically Relevant Mutations in Liquid Biopsies of Cancer Patients Using a Hybridization-Based Platform

BACKGROUND: With the advent of precision oncology, liquid biopsies are quickly gaining acceptance in the clinical setting. However, in some cases, the amount of DNA isolated is insufficient for Next-Generation Sequencing (NGS) analysis. The nCounter platform could be an alternative, but it has never been explored for detection of clinically relevant alterations in fluids. METHODS: Circulating-free DNA (cfDNA) was purified from blood, cerebrospinal fluid, and ascites of patients with cancer and analyzed with the nCounter 3 D Single Nucleotide Variant (SNV) Solid Tumor Panel, which allows for detection of 97 driver mutations in 24 genes. RESULTS: Validation experiments revealed that the nCounter SNV panel could detect mutations at allelic fractions of 0.02-2% in samples with 5 pg mutant DNA/mL. In a retrospective analysis of 70 cfDNAs from patients with cancer, the panel successfully detected EGFR, KRAS, BRAF, PIK3CA, and NRAS mutations when compared with previous genotyping in the same liquid biopsies and paired tumor tissues [Cohen kappa of 0.96 (CI = 0.92-1.00) and 0.90 (CI = 0.74-1.00), respectively]. In a prospective study including 91 liquid biopsies from patients with different malignancies, 90 yielded valid results with the SNV panel and mutations in EGFR, KRAS, BRAF, PIK3CA, TP53, NFE2L2, CTNNB1, ALK, FBXW7, and PTEN were found. Finally, serial liquid biopsies from a patient with NSCLC revealed that the semiquantitative results of the mutation analysis by the SNV panel correlated with the evolution of the disease. CONCLUSIONS: The nCounter platform requires less DNA than NGS and can be employed for routine mutation testing in liquid biopsies of patients with cancer

Multiplex RNA-based detection of clinically relevant MET alterations in advanced non-small cell lung cancer

MET inhibitors have shown activity in non-small-cell lung cancer patients (NSCLC) with MET amplification and exon 14 skipping (METΔex14). However, patient stratification is imperfect, and thus, response rates have varied widely. Here, we studied MET alterations in 474 advanced NSCLC patients by nCounter, an RNA-based technique, together with next-generation sequencing (NGS), fluorescence in situ hybridization (FISH), immunohistochemistry (IHC), and reverse transcriptase polymerase chain reaction (RT-PCR), exploring correlation with clinical benefit. Of the 474 samples analyzed, 422 (89%) yielded valid results by nCounter, which identified 13 patients (3%) with METΔex14 and 15 patients (3.5%) with very-high MET mRNA expression. These two subgroups were mutually exclusive, displayed distinct phenotypes and did not generally coexist with other drivers. For METΔex14, 3/8 (37.5%) samples positive by nCounter tested negative by NGS. Regarding patients with very-high MET mRNA, 92% had MET amplification by FISH and/or NGS. However, FISH failed to identify three patients (30%) with very-high MET RNA expression, among which one received MET tyrosine kinase inhibitor treatment deriving clinical benefit. Our results indicate that quantitative mRNA-based techniques can improve the selection of patients for MET-targeted therapies

Anales de Edafología y Fisiología Vegetal Tomo 16

Nuevo reactivo para la determinación calorimétrica del fósforo en suelos, l. Propiedades generales, por F. Lucena Conde y L. Prat.-- Nuevo reactivo para la determinación calorimétrica del fósforo en suelos, II Aplicación al análisis de suelos, por F. Lucena Conde y L. Prat.-- Aplicación del índice de los minerales básicos para deducir la fertilidad de los suelos, en los trópicos, por Josefina Pérez Mateos.-- Contribución al estudio de la fertilidad de los suelos de la Huerta de Murcia, por O. Carpena, C. Abrisqueta, M. G. Guillén y J. A. Sánchez.-- Composición químiCa de hojas de Citrus. I. Limonero, por O. Carpena, C. Abrisqueta, J. A. Sánchez y M. G. Guillén.-- Estudios edafológico-geográficos acerca de las formaciones edáficas sobre roca caliza de España meridional y sur-oriental, por H. Klinge.-- Síntesis del butadieno a partir del alcohol etílico influencia de catalizadores arcilla-MgO, por Vicente Aleixandre y Teófilo Fernández.-- Aplicaciones ánalíticas de la fotometría de llama. I. Estudio de las interferencias mutuas en las determinaciones de Na, K y Ca en extractos de suelo obtenidos con acetato amónico N, por F. Burriel-Martí, J. Ramírez-Muñoz y Adelaida Benito-Potous.-- Contribución al conocimiento de los suelos recientes, relictos y fósiles sobre roca caliza, del Norte de España, por H . Klínge.-- La clorosis férrica del limonero l, por O. Carpena, M. G. Guillén y J. A. Sánchez.-- La clorosis férrica del limonero II por O. Carpena, J. A. Sánchez y M. G. Guillén. Estudio de tierras pardas y ranker canarios en relación con la naturaleza de la roca madre, por Angel Hoyos de Castro y Vicente Soler.-- Acerca de la existencia de un sistema de defensa celular durante la fase de crecimiento: citoarjesis e hipótesis de Ios citoarjés, por Román Vicente Jordana.-- Los arenales costeros del Levante español I. Las playas de Valencia. Estudio mineralógico de suis materiales. por Josefina Pérez Mateas y Juan J. Alonso Pascual.-- La red fluvial de Valencia. II. Mitad norte de la provincia, por Juan J. Alonso Pascual.-- El agente etiológico de la tuberculosis del olivo en relación con el suelo, por R. Beltrá.-- Los españoles de la zona árida formados sobre sedimentos silúricos, por José María Albareda, Vicente Aleixandre y Mª del Carmen Sánchez Calvo.-- Aplicación de las medidas de viscosidad absoluta al estudio de la acción de los defloculantes sobre las suspensiones acuosas de· minerales arcillosos, por L. Rodrigo, F. Costell y J. l. Fenández Alonso.-- Construcción y funcionamiento de un sencillo modelo de Fotómetro de llama, por José Cardús Aguilar.-- Fertilización del Dianthus caryophyllus en cultivos industriales I por José Cardús, y Jorge F. Aguilá.-- Contribución a la ecología de los tardígrados de suelos húmedos. por F. Michelcic.-- La red fluvial de Valencia. III. Mitad sur de la provincia. Conclusiones, por Juan J. Alonso Pascual.-- Influencia de algunos factores sobre la aparición de cierto grado de fitotoxicidad en la naranja W Navel frente al uso del ortofenilfenato sódico-hexamina, por Benito Orihuel Gasqué y José Merí Puig.-- Génesis de la montmorillonita de Marruecos español, II, por E. Gutiérrez, Ríos, J. L. Martín Vivaldi y C. del Pino Vázquez.--Modificación de las propiedades adsorbentes de los minerales arcillosos mediante tratamiento térmico y catión de cambio, I, por Vicente Aleixandre y Mª Covadonga Rodríguez.-- Clasificación filogenética y ecológica de los suelos del mundo, por Manuel del Llano.-- Auxinas e inhibidores del crecimiento en las plantas, por Ramiro Díaz Eimil.-- Estudio citológico de la epidermis del bulbo de una variedad hexaploide de Scilla marítima L. por M. Losada y G. Giménez.-- Composición mineralógica y génesis de algunos tipos de suelos calizos. béticos. l. Mineralogía de las fracciones gruesas {arenas), por G. Paneque Guerrero y F. González García.-- Sobre la determinación de fósforo y potasio asimilables en los suelos de la Vega de Granada. l. Extracción del fósforo mediante la mezcla acético-acetato, amónico, por F. Capitán-García y R. García-Ruiz.-- Estudio acerca de la influencia de diferentes materiales orgánicos sobre el nivel de nitrógeno y pH de los suelos alcalinos, por S. P. Mitra y Hari Shanker.-- Los efectos del estilbestrol por vía ·orar sobre la retención de nitrógeno y la digestibilidad en corderos enteros y Castrados, por Eduardo Zorita y Gaspar González.-- El encalado de los suelos de zona húmeda, por F. Guitián Ojea y M. Muñoz Taboadela.--La lana del vidrio como soporte del cultivo en el estudio sobre las liciencias del boro en plantas, por F. Lucena Conde y R. Aragonés Apodaca.-- Composición mineralógica y génesis de algunos tipos de suelos calizos béticos. II. Estudio fisico-químico y mineralógico de las fracciones finas (arcillas), por F. González García y G. Paneque Guerrero.-- Estudio fisicoquimico de las arcillas del Levante espaiñol. V. Ensayos técnicos, por A. Grua Mestre, F. Costell Landete y J. I. Fernández Alonso.-- El problema de las bioformas de los tardígrados, por Franc Milhelcic. Información. Notas. Profs. J. Mitchell y P. Ehrenberg.-- Notas Congreso de la U. I. de Química Pura y Aplicada. Notas. Nueva sección. Sociedad Española de Ciencia del Suelo.-- Notas. Nuevos Colaboradores.-Trabajo premiado. Notas. LI Asamblea del Frío. -- Notas. Prof. A .. Boerger. 117 aniversario de la Facultad de Agricultura de Pisa. V Congreso Internacional de la IQUA. Notas. Conferencia del prof. Klotz. Congreso para la protección de las plantas.-Congreso de Bioquímica.—Notas. Coloquio. Pensionado. Doctorados.-- Notas. Sociedad Española de la Ciencia del Suelo.-- Bibliografía. D. W. Thorne y H. R. Peterson: I rigated soils. Their fertility and menagement. Handbuch der Pflanzenphysilogie. Vol. I. XV Congrès International de Chimíe puré et appliquée. Experientia .Supplementum V.-- H. Moritz: Spectrochemische Betriebanalyse. C. R. Strouts. J. H. Gilfillan and N. H. Wilson (Ed.): Analytical Chernistry. The Working Tools.-- Walter Wagner, Clarence J. Hull and Gerald E. Markle : Avanced Analytical Chemistry.-- Instituto Internacional de la potasa. Potassium Symposiumn 1956.-- S. H. Stallings: Soil Conservation.-- Samuel L. Tisdale and Werner L. Nelsotl : Soil fertility and fertilizersPeer reviewe

BID expression determines the apoptotic fate of cancer cells after abrogation of the spindle assembly checkpoint by AURKB or TTK inhibitors

Background: Drugs targeting the spindle assembly checkpoint (SAC), such as inhibitors of Aurora kinase B (AURKB) and dual specific protein kinase TTK, are in different stages of clinical development. However, cell response to SAC abrogation is poorly understood and there are no markers for patient selection. Methods: A panel of 53 tumor cell lines of different origins was used. The effects of drugs were analyzed by MTT and flow cytometry. Copy number status was determined by FISH and Q-PCR; mRNA expression by nCounter and RT-Q-PCR and protein expression by Western blotting. CRISPR-Cas9 technology was used for gene knock-out (KO) and a doxycycline-inducible pTRIPZ vector for ectopic expression. Finally, in vivo experiments were performed by implanting cultured cells or fragments of tumors into immunodeficient mice. Results: Tumor cells and patient-derived xenografts (PDXs) sensitive to AURKB and TTK inhibitors consistently showed high expression levels of BH3-interacting domain death agonist (BID), while cell lines and PDXs with low BID were uniformly resistant. Gene silencing rendered BID-overexpressing cells insensitive to SAC abrogation while ectopic BID expression in BID-low cells significantly increased sensitivity. SAC abrogation induced activation of CASP-2, leading to cleavage of CASP-3 and extensive cell death only in presence of high levels of BID. Finally, a prevalence study revealed high BID mRNA in 6% of human solid tumors. Conclusions: The fate of tumor cells after SAC abrogation is driven by an AURKB/ CASP-2 signaling mechanism, regulated by BID levels. Our results pave the way to clinically explore SAC-targeting drugs in tumors with high BID expression.Fil: Bertran Alamillo, Jordi. Quiron Dexeus University Hospital; EspañaFil: Giménez Capitán, Ana. Quiron Dexeus University Hospital; EspañaFil: Román, Ruth. Quiron Dexeus University Hospital; EspañaFil: Talbot, Sara. Quiron Dexeus University Hospital; EspañaFil: Whiteley, Rebecca. Quiron Dexeus University Hospital; EspañaFil: Floch, Nicolas. Astrazeneca; Reino UnidoFil: Martinez Perez, Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Martin, Matthew J.. Astrazeneca; Reino UnidoFil: Smith, Paul D.. Astrazeneca; Reino UnidoFil: Sullivan, Ivana. Hospital de la Santa Creu I Sant Pau; España. Hospital Universitari Dexeus; EspañaFil: Terp, Mikkel G.. University of Southern Denmark; DinamarcaFil: Saeh, Jamal. Astrazeneca; Reino UnidoFil: Marino Buslje, Cristina. Fundación Instituto Leloir; ArgentinaFil: Fabbri, Giulia. Astrazeneca; Reino UnidoFil: Guo, Grace. Astrazeneca; Reino UnidoFil: Xu, Man. Astrazeneca; Reino UnidoFil: Tornador, Cristian. Teresa Moretó Foundation And Wholegenix Sl; EspañaFil: Aguilar Hernández, Andrés. Hospital Universitari Dexeus; EspañaFil: Reguart, Noemí. Hospital Clinic Barcelona; EspañaFil: Ditzel, Henrik J.. University of Southern Denmark,; Dinamarca. Odense University Hospital; DinamarcaFil: Martínez Bueno, Alejandro. Hospital Universitari Dexeus; EspañaFil: Nabau Moretó, Núria. Teresa Moretó Foundation And Wholegenix Sl; EspañaFil: Gascó, Amaya. Astrazeneca; Reino UnidoFil: Rosell, Rafael. Germans Trias I Pujol Research Institute; España. Hospital Universitari Dexeus; EspañaFil: Pease, J. Elizabeth. Astrazeneca; Reino UnidoFil: Polanska, Urszula M.. Astrazeneca; Reino UnidoFil: Travers, Jon. Astrazeneca; Reino UnidoFil: Urosevic, Jelena. Astrazeneca; Reino UnidoFil: Molina Vila, Miguel A.. Hospital Universitari Dexeus; Españ