Phospho-RNA-Seq Highlights Specific Small RNA Profiles in Plasma Extracellular Vesicles.

Small RNAs (sRNAs) are bioactive molecules that can be detected in biofluids, reflecting physiological and pathological states. In plasma, sRNAs are found within extracellular vesicles (EVs) and in extravesicular compartments, offering potential sources of highly sensitive biomarkers. Deep sequencing strategies to profile sRNAs favor the detection of microRNAs (miRNAs), the best-known class of sRNAs. Phospho-RNA-seq, through the enzymatic treatment of sRNAs with T4 polynucleotide kinase (T4-PNK), has been recently developed to increase the detection of thousands of previously inaccessible RNAs. In this study, we investigated the value of phospho-RNA-seq on both the EVs and extravesicular plasma subfractions. Phospho-RNA-seq increased the proportion of sRNAs used for alignment and highlighted the diversity of the sRNA transcriptome. Unsupervised clustering analysis using sRNA counts matrices correctly classified the EVs and extravesicular samples only in the T4-PNK treated samples, indicating that phospho-RNA-seq stresses the features of sRNAs in each plasma subfraction. Furthermore, T4-PNK treatment emphasized specific miRNA variants differing in the 5'-end (5'-isomiRs) and certain types of tRNA fragments in each plasma fraction. Phospho-RNA-seq increased the number of tissue-specific messenger RNA (mRNA) fragments in the EVs compared with the extravesicular fraction, suggesting that phospho-RNA-seq favors the discovery of tissue-specific sRNAs in EVs. Overall, the present data emphasizes the value of phospho-RNA-seq in uncovering RNA-based biomarkers in EVs.

Plasma-derived FVIII/VWF complex shows higher protection against inhibitors than isolated FVIII after infusion in haemophilic patients: A translational study.

INTRODUCTION: Presence of von Willebrand factor (VWF) in FVIII concentrates offers protection against neutralizing inhibitors in haemophilia A (HA). Whether this protection is more evident in plasma-derived (pd) FVIII/VWF or recombinant (r) FVIII concentrates remains controversial. AIM: We investigated the protection exerted by VWF against FVIII inhibitors in an in vivo mouse model of HA exposed to pdFVIII/VWF or to various rFVIII concentrates. METHODS: Haemophilia A mice received the different FVIII concentrates after administration of vehicle or an inhibitory IgG purified from a commercial pool of HA plasma with inhibitors and FVIII:C recoveries were measured. Furthermore, using a novel clinically oriented ex vivo approach, Bethesda inhibitory activities (BU) of a commercial pool of HA plasma with inhibitors were assessed using normal plasma, or plasma from severe HA patients, without inhibitors, after treatment with the same concentrates. RESULTS: in vivo studies showed that pdFVIII/VWF offers markedly higher protection against inhibitors when compared with any of the FVIII products without VWF. More importantly, in the ex vivo studies, plasma from patients treated with pdFVIII/VWF showed higher protection against inhibitors (P values ranging .05-.001) in comparison with that observed in plasma from patients who received FVIII products without VWF, regardless of the type of product evaluated. CONCLUSION: Data indicate that FVIII+VWF complexes assembled in the circulation after rFVIII infusion are not equivalent to the naturally formed complex in pdFVIII/VWF. Therefore, rFVIII infused into HA patients with inhibitors would be less protected by VWF than the FVIII in pdFVIII/VWF concentrates.

Plasma exchange with albumin replacement for Alzheimer's disease treatment induced changes in serum and cerebrospinal fluid inflammatory mediator levels.

OBJECTIVE: There is extensive literature indicating that inflammatory pathways are affected in Alzheimer's disease (AD). We examined whether plasma exchange with albumin replacement (PE-Alb) can impact the inflammatory status of AD patients and alter the relationship between inflammatory mediators and cognitive measures. METHODS: Serum and cerebrospinal fluid (CSF) samples from 142 AD patients participating in the AMBAR trial (14-month schedule of PE-Alb treatment vs. placebo [sham PE-Alb]) were analyzed for changes from baseline for 19 inflammatory mediators (6 inflammatory cytokines, 9 chemokines, and 4 vascular injury indicators) at representative time points across the AMBAR study (lasting effects) as well as in pre- versus post-PE-Alb procedure (acute effects). Association between mediator changes and clinical outcomes reported in the AMBAR study (cognitive, functional, behavioral function, and global change tests) was assessed. RESULTS: PE-Alb significantly reduced IFN-γ, eotaxin, MIP-1α and ICAM-1 levels in serum, and eotaxin-3 and MIP-1ß levels in CSF, at various time points during treatment (p < 0.05; false discovery rate-corrected). Vascular injury indicators were the mediators mostly affected by post- versus pre-PE-Alb level reduction. Increased serum MIP-1α levels were associated with worsening in ADAS-Cog, CDR-sb, and ADCS-CGIC scores in the placebo group, but not in the PE-Alb-treated group. INTERPRETATION: Peripheral intervention could affect AD by reducing inflammatory mediators in both peripheral and central compartments. Changes in MIP-1α due to PE-Alb were associated with changes in clinical outcomes.

Non-additive effect on thrombin generation when a plasma-derived factor VIII/von Willebrand factor (FVIII/VWF) is combined with emicizumab in vitro.

BACKGROUND: Emicizumab is an alternative non-factor approach for treating patients with hemophilia A. However, there is a potential risk of thrombotic events when emicizumab is concomitantly administered with pro-hemostatic therapies. OBJECTIVES: To assess the hemostatic effect in vitro when a plasma-derived factor VIII concentrate containing von Willebrand factor (pdFVIII/VWF) was added to hemophilia A plasma (HAp) in combination with emicizumab. METHODS: HAp and HAp with FVIII inhibitors (HAp-i) samples with different concentrations of emicizumab (50 and 100 µg/mL) were combined with activated prothrombin complex concentrate at 0.5 to 1 U/mL, recombinant activated factor VII (rFVIIa) at 0.5 to 7 µg/mL, and pdFVIII/VWF at 0.1 to 4.5 IU/mL. Thrombin generation (TG; thrombin peak and endogenous thrombin potential) was determined using a Calibrated Automated Thrombogram assay. RESULTS: When activated prothrombin complex concentrate was added to HAp and HAp-i with emicizumab, TG dramatically increased (multiplier effect > 4.5×). Addition of rFVIIa to HAp or HAp-i with emicizumab moderately increased TG in a concentration-related manner compared with rFVIIa alone. Addition of pdFVIII/VWF to HAp or HAp-i with emicizumab induced a TG response equivalent to those samples without emicizumab. In an in vitro model of immune tolerance induction with bleeds (HAp-i 15 Bethesda units), combination of pdFVIII/VWF, emicizumab, and rFVIIa did not trigger a multiplying effect on TG. CONCLUSIONS: pdFVIII/VWF showed a non-additive effect on TG when combined in vitro with emicizumab. This finding suggests that emicizumab has limited ability to promote factor X activation in the presence of pdFVIII/VWF, thus reducing the risk of thrombotic events.

Predicting Job Satisfaction in Military Organizations: Unpacking the Relationship Between Emotional Intelligence, Teamwork Communication, and Job Attitudes in Spanish Military Cadets.

Although prior research has extensively examined the association of emotional intelligence (EI) with various job attitudes (e.g., job satisfaction), empirical and systematic investigation of this link within military institutions has captured considerably less attention. The present research analyzed the relationship between EI, teamwork communication, and job satisfaction among Spanish military cadets. We tested the potential unique contribution of EI to job satisfaction over and above demographics (i.e., gender and age), proactive personality, and resilience. Moreover, we also examined whether EI was indirectly linked to job satisfaction via its relationship with teamwork communication. A sample of 363 cadet officers of the Spanish General Military Academy completed questionnaires assessing EI, teamwork communication, proactive personality, resilience, and job satisfaction. Hierarchical regression analysis revealed that EI exhibited incremental variance (ΔR 2 = 5.2%) in predicting job satisfaction (B = 0.539, 95% CI [0.306,0.771]) even after accounting for demographics, proactive personality, and resilience. Additionally, mediation analysis showed that the association of EI with job satisfaction was partially driven by enhanced teamwork communication. This research provides empirical evidence suggesting a pathway (i.e., effective teamwork communication) through which EI could help military cadets to experience higher job satisfaction. Implications for future academic programs including EI and teamwork communication to promote positive job attitudes among military personnel are discussed.

Modulation of genotoxicity and cytotoxicity by radish grown in metal-contaminated soils.

Members of the Brassicaceae family are known for their anticarcinogenic and genetic material protective effects. However, many of the species of this family accumulate high amounts of metals, which is an undesirable feature. Radish (Raphanus sativus L.) has shown to accumulate metals in roots to a higher extent than others members of Brassicaceae. The main objectives of this work are (i) to study the distribution of the accumulated As, Pb and Cd in radish plants and (ii) to establish the genotoxic, antigenotoxic and cytotoxic activities of the root and shoot of this vegetable. Results indicate that (i) the shoots of radish accumulate higher concentrations of metal(oid)s than roots; (ii) the shoots were genotoxic at the different concentrations studied, with the root showing such genotoxic effect only at the highest concentration assayed; (iii) the antigenotoxic potential of radish is reduced in plants with high metal content and (iv) the tumouricide activities of the radish plants were negatively correlated to their metal(oid) contents. An interaction between metal(oid)s and the isotyocianates (hydrolysis products of the glucosinolates) contained in the radish is suggested as the main modulator agents of the genotoxic activity of the plants grown in contaminated soils with metal(oid)s.

Nivel de conocimientos y capacidad para el autocuidado en personas en tratamiento con hemodiálisis y su relación con la calidad de vida / Level of knowledge and self-care capacity in individuals undergoing hemodialysis and its relationship with quality of life

Introducción:Determinar el grado de conocimientos que tie-nen los pacientes en hemodiálisis sobre su enfermedad y trata-miento, valorar su capacidad para hacerse el autocuidado y su relación con la calidad de vida relacionada con la salud.Material y Método: Se realizó un estudio descriptivo y trans-versal en dos unidades de diálisis. Se estudiaron 31 pacien-tes en hemodiálisis con edad media de 67,2±14 años (71% hombres). Se estudió nivel de conocimientos sobre la enfermedad y tra-tamiento, capacidad de autocuidado y calidad de vida (lámi-nas COOP/WONCA). Se recogieron, además, variables socio-demográficas, clínicas, comorbilidad y grado de dependencia.Resultados: El 9,7% de los pacientes eran laboralmente acti-vos. El 16,1% no tenía formación, 38,7% básica, 32,3% secun-daria y 12,9% universitaria. El 61% tenía un grado de conoci-mientos alto sobre su enfermedad y tratamiento, el 32% medio y el 7% bajo.El 52 % de los pacientes mostraron alta capacidad para el au-tocuidado y el 48% media. El 3,2% de los pacientes presentaba dependencia severa, moderada el 22,6%, leve el 6,5%, siendo autónomos el 67,7%.La puntuación media en el cuestionario de calidad de vida fue de 24,06±6,5 puntos.Encontramos relación significativa entre el grado de conocimien-tos con edad (0,384, p<0,05) y nivel formativo (0,464, p<0,01).Conclusiones: Los pacientes presentan un alto nivel de cono-cimiento sobre su enfermedad y tratamiento, alto grado de autonomía y capacitación para el autocuidado, siendo los más jóvenes y con mayor nivel formativo, los que muestran mejores resultados, sin que estas variables tengan relación con la cali-dad de vida (AU)

Introduction:To determine the level of knowledge that hemodialysis patients have about their disease and treatment, assess their ability for self-care, and examine its relationship with health-related quality of life.Material and Method:A descriptive, cross-sectional study was conducted in two dialysis units. Thirty-one hemodialysis patients were studied, with a mean age of 67.2±14 years (71% male). The study assessed the level of knowledge about the disease and treatment, self-care ability, and quality of life (using COOP/WONCA charts). Additionally, sociodemographic, clinical, comorbidity, and dependence variables were collected.Results: 9.7% of the patients were employed. 16.1% had no formal education, 38.7% had basic education, 32.3% had secondary education, and 12.9% had a university education. 61% had a high level of knowledge about their disease and treatment, 32% had a moderate level, and 7% had a low level.52% of the patients demonstrated a high capacity for self-care, while 48% showed a moderate capacity. 3.2% of the patients exhibited severe dependence, 22.6% moderate dependence, 6.5% mild dependence, and 67.7% were autonomous. The average score on the quality-of-life questionnaire was 24.06±6.5 points. A significant relationship was found between the level of knowledge and age (0.384; p<0.05) and educational level (0.464; p<0.01).Conclusions: Patients exhibit a high level of knowledge about their disease and treatment, along with a high degree of autonomy and self-care capabilities. Younger patients and those with higher education levels show better results, although these variables do not appear to be directly related to quality of life (AU)

Cancer, Warts, or Asymptomatic Infections: Clinical Presentation Matches Codon Usage Preferences in Human Papillomaviruses.

Viruses rely completely on the hosts' machinery for translation of viral transcripts. However, for most viruses infecting humans, codon usage preferences (CUPrefs) do not match those of the host. Human papillomaviruses (HPVs) are a showcase to tackle this paradox: they present a large genotypic diversity and a broad range of phenotypic presentations, from asymptomatic infections to productive lesions and cancer. By applying phylogenetic inference and dimensionality reduction methods, we demonstrate first that genes in HPVs are poorly adapted to the average human CUPrefs, the only exception being capsid genes in viruses causing productive lesions. Phylogenetic relationships between HPVs explained only a small proportion of CUPrefs variation. Instead, the most important explanatory factor for viral CUPrefs was infection phenotype, as orthologous genes in viruses with similar clinical presentation displayed similar CUPrefs. Moreover, viral genes with similar spatiotemporal expression patterns also showed similar CUPrefs. Our results suggest that CUPrefs in HPVs reflect either variations in the mutation bias or differential selection pressures depending on the clinical presentation and expression timing. We propose that poor viral CUPrefs may be central to a trade-off between strong viral gene expression and the potential for eliciting protective immune response.