Symptomatic carotid atherosclerotic plaques are associated with increased infiltration of natural killer (NK) cells and higher serum levels of NK activating receptor ligands

A wide array of immune cells, including lymphocytes, is known to be present and to play a pathogenetic role in atherosclerotic lesions. However, limited information is currently available regarding the presence of Natural Killer (NF cell subsets within vessel plaque, and more in general, regarding their role in human atherosclerosis. We evaluated the distribution of NK cells in human carotid atherosclerotic plaques, dissecting asymptomatic and symptomatic patients (identified as affected by stroke, transient ischemic attack, or amaurosis fugax within 6 months) with the aim of shedding light on the putative contribution of NK cells to the pathogenic process that leads to plaque instability and subsequent clinical complications. We observed that carotid plaques were consistently infiltrated by NK cells and, among them, CD56(bright)perforin(low) NK cells were abundantly present and displayed different markers of tissue residency (i.e., CD103 CD69 and CD49a). Interestingly, carotid atherosclerotic plaques of symptomatic patients showed a higher content of NK cells and an increased ratio between CD56(bright)perforin(low) NK cells and their CD56(dim)perforin(high)counterpart. NK cells isolated from plaques of symptomatic patients were also stronger producers of IFN-gamma. Analysis of the expression of NK activating receptor ligands (including MICA/B, ULBP-3, and B7-H6) in atherosclerotic carotid plaques revealed that they were abundantly expressed by a HLA-DR(+)CD11c(+) myeloid cell population resident in the plaques. Remarkably, sera of symptomatic patients contained significant higher levels of soluble ligands for NK activating receptors. Our observations indicate that CD56(bright)( )NK cells accumulate within human atherosclerotic lesions and suggest a possible contribution of NK cells to the process determining plaque instability

Acute atrial ischemia associates with early but not late new-onset atrial fibrillation in STEMI patients treated with primary PCI: relationship with in-hospital outcomes

Abstract Funding Acknowledgements Type of funding sources: None. OnBehalf N/A Background. New-onset atrial fibrillation (NOAF) is known to be a common complication in STEMI patients undergoing primary percutaneous coronary intervention (PCI), which is associated with a negative short- and long-term prognosis. Recently, two distinct phenotypes of NOAF have been described, namely early (EAF) and late NOAF (LAF). However, whether EAF and LAF recognize different pathogenetic mechanisms is unknown. Purpose. To investigate atrial branches occlusion and EAF or LAF onset in STEMI patients undergoing primary PCI. Methods. Retrospective cohort study including 155 STEMI patients. Patients were divided into 3 groups: sinus rhythm (SR), EAF or LAF. Clinical characteristics, angiographic features including occlusion of atrial branches, namely ramus ostia cavae superioris (ROCS), atrio-ventricular node artery (AVNA), right intermediate atrial artery (RIAA) and left intermediate atrial artery (LIAA), were assessed. We also investigated in-hospital complications, death, and a composite of major post-NOAF adverse events (AEs) including cardiogenic shock, acute pulmonary edema, sustained ventricular tachycardia and ventricular fibrillation. Results. Mean age was 63.8 ± 11.9 years; 78.7% of men. NOAF was detected in 22 (14.2%) patients: 10 (6.4%) EAF and 12 LAF (7.7%). Compared to EAF, LAF patients were older (p = 0.013), with higher GRACE risk score (p = 0.014) and Killip class (p = 0.015), depressed ejection fraction (p = 0.007), elevated filling pressures (p = 0.029), higher c-reactive protein (p = 0.014) and more TIMI flow <3 (p = 0.015). As shown in Figure 1, EAF was associated with higher prevalence of occluded ROCS (p = 0.010), AVNA (p = 0.005) and RIAA (p < 0.001), compared to SR. Moreover, EAF patients had more frequently ≥2 diseased atrial branches than SR (19.5%, p < 0.001) and LAF (25%, p < 0.030) patients. In LAF patients, a higher incidence of pre-PCI cardiogenic shock, post-PCI AEs (p = 0.019 vs SR; p = 0.029 vs EAF) and death (p = 0.004 vs SR) was found. Conclusions. The occlusion of atrial branches is associated with early but not late NOAF following STEMI. LAF patients had worse in-hospital AEs and mortality. Abstract Figure

B-cell-specific checkpoint molecules that regulate anti-tumour immunity.

The role of B cells in anti-tumour immunity is still debated and, accordingly, immunotherapies have focused on targeting T and natural killer cells to inhibit tumour growth1,2. Here, using high-throughput flow cytometry as well as bulk and single-cell RNA-sequencing and B-cell-receptor-sequencing analysis of B cells temporally during B16F10 melanoma growth, we identified a subset of B cells that expands specifically in the draining lymph node over time in tumour-bearing mice. The expanding B cell subset expresses the cell surface molecule T cell immunoglobulin and mucin domain 1 (TIM-1, encoded by Havcr1) and a unique transcriptional signature, including multiple co-inhibitory molecules such as PD-1, TIM-3, TIGIT and LAG-3. Although conditional deletion of these co-inhibitory molecules on B cells had little or no effect on tumour burden, selective deletion of Havcr1 in B cells both substantially inhibited tumour growth and enhanced effector T cell responses. Loss of TIM-1 enhanced the type 1 interferon response in B cells, which augmented B cell activation and increased antigen presentation and co-stimulation, resulting in increased expansion of tumour-specific effector T cells. Our results demonstrate that manipulation of TIM-1-expressing B cells enables engagement of the second arm of adaptive immunity to promote anti-tumour immunity and inhibit tumour growth

Crystal structure and centromere binding of the plasmid segregation protein ParB from pCXC100

Plasmid pCXC100 from the Gram-positive bacterium Leifsonia xyli subsp. cynodontis uses a type Ib partition system that includes a centromere region, a Walker-type ATPase ParA and a centromere-binding protein ParB for stable segregation. However, ParB shows no detectable sequence homology to any DNA-binding motif. Here, we study the ParB centromere interaction by structural and biochemical approaches. The crystal structure of the C-terminal DNA-binding domain of ParB at 1.4 Å resolution reveals a dimeric ribbon–helix–helix (RHH) motif, supporting the prevalence of RHH motif in centromere binding. Using hydroxyl radical footprinting and quantitative binding assays, we show that the centromere core comprises nine uninterrupted 9-nt direct repeats that can be successively bound by ParB dimers in a cooperative manner. However, the interaction of ParB with a single subsite requires 18 base pairs covering one immediate repeat as well as two halves of flanking repeats. Through mutagenesis, sequence specificity was determined for each position of an 18-bp subsite. These data suggest an unique centromere recognition mechanism by which the repeat sequence is jointly specified by adjacent ParB dimers bound to an overlapped region

The interaction of Pcf11 and Clp1 is needed for mRNA 3′-end formation and is modulated by amino acids in the ATP-binding site

Polyadenylation of eukaryotic mRNAs contributes to stability, transport and translation, and is catalyzed by a large complex of conserved proteins. The Pcf11 subunit of the yeast CF IA factor functions as a scaffold for the processing machinery during the termination and polyadenylation of transcripts. Its partner, Clp1, is needed for mRNA processing, but its precise molecular role has remained enigmatic. We show that Clp1 interacts with the Cleavage–Polyadenylation Factor (CPF) through its N-terminal and central domains, and thus provides cross-factor connections within the processing complex. Clp1 is known to bind ATP, consistent with the reported RNA kinase activity of human Clp1. However, substitution of conserved amino acids in the ATP-binding site did not affect cell growth, suggesting that the essential function of yeast Clp1 does not involve ATP hydrolysis. Surprisingly, non-viable mutations predicted to displace ATP did not affect ATP binding but disturbed the Clp1–Pcf11 interaction. In support of the importance of this interaction, a mutation in Pcf11 that disrupts the Clp1 contact caused defects in growth, 3′-end processing and transcription termination. These results define Clp1 as a bridge between CF IA and CPF and indicate that the Clp1–Pcf11 interaction is modulated by amino acids in the conserved ATP-binding site of Clp1

Habilidades funcionales de niños, niñas y adolescentes con parálisis cerebral y su relación con el compromiso motor y la discapacidad intelectual en Argentina.

Introducción: El compromiso motor caracteriza la parálisis cerebral (PC), y suele asociarse a la discapacidad intelectual (DI). Se han desarrollado sistemas de clasificación estandarizados para describir las funciones de personas con PC. Objetivo: caracterizar funcionalmente a niños, niñas y adolescentes de 0 a 18 años con PC de Argentina e indagar la asociación entre el compromiso motor (GMFCS), la DI y las clasificaciones funcionales. Métodos: Estudio transversal. Se recolectaron datos a través de entrevistas a las familias y revisión de historias clínicas. Se incluyeron personas con PC. Los datos se recolectaron de 19 instituciones de distintas ciudades de Argentina. Para el análisis de los datos se utilizó test de Fisher y odds ratio [IC95%], con significación <0,05. Resultados: participaron 182 niños, niñas y adolescentes con PC. Según clasificación GMFCS prevaleció el nivel V con 36,3%. Quienes presentan compromiso motor más severo (GMFCS IV-V), tienen 72 [25,4;206,0] veces y 13 [5,9;28,2] veces más chances de presentar un nivel severo de MACS y CFCS respectivamente. Pero, presentaron 34 [7,9;146,0] veces más chances de un nivel leve a moderado de EDACS. Quienes presentaron DI tuvieron 10 [5,1;20,5] veces más chances de presentar un nivel severo GMFCS, 6 [3,4;13,2] veces más chances un nivel severo MACS y 4 [2,0;7,8] veces más chances de un nivel severo CFCS. Por el contrario, tienen 4 [1,9;9,5] veces más chances de presentar un nivel leve-moderado EDACS. Conclusión: el nivel de GMFCS y la presencia de DI influyen en la funcionalidad general y aumentan la severidad en el compromiso, habilidades manuales y de comunicación

The structure of the tetrasialoganglioside from human brain

Autosomal dominant retinal vasculopathy with cerebral leukodystrophy is a microvascular endotheliopathy with middle- age onset. In nine families, we identified heterozygous C- terminal frameshift mutations in TREX1, which encodes a 3'-5' exonuclease. These truncated proteins retain exonuclease activity but lose normal perinuclear localization. These data have implications for the maintenance of vascular integrity in the degenerative cerebral microangiopathies leading to stroke and dementias

Understanding EU legal integration/disintegration : in search of new perspectives

This report summarises the UACES/ James Madison Trust EUFutures Research Network Launch Workshop entitled 'Understanding legal integration/disintegration: in search of new perspectives'. The event consisted of four panels on 'Interdisciplinary research on EU law', 'Research Methods and EU law', 'Understanding the EU's integration processes' and 'Understanding EU law through soft law, discourse, ideas & beliefs', respectively. The future of EU legal integration is at a significant juncture with the departure of the UK, substantial rule of law challenges, internal and external crises, and an increasingly apathetic multilateral legal order. There is increased recognition amongst EU lawyers, who have historically limited themselves to doctrinal analysis and legal hermeneutics, that methodology plays an essential role in order to understand EU integration and shape its future. The question remains though how to connect interdisciplinary approaches to EU law, policy and politics. How should EU law (as an object) be studied? What are the respective merits of each discipline (political science, sociology, economy, history) in explaining the way EU law is created, applied, used, transformed in the process of EU integration? What is the added value of bringing together different approaches to law? In particular, how can EU law (as an academic discipline) open itself up to the methods of the social sciences and what, in return, can law offer to our understanding of EU studies more widely? In order to answer these questions, EUFutures brings together scholars for this workshop to: reflect on the future methodological direction(s) of EU law and EU integration and consider both how law could open itself up to methodologies from other disciplines, and what legal analysis could offer political, economic and historical approaches

Insight from an Italian Delphi Consensus on EVAR feasibility outside the instruction for use: the SAFE EVAR Study

BACKGROUND: The SAfety and FEasibility of standard EVAR outside the instruction for use (SAFE-EVAR) Study was designed to define the attitude of Italian vascular surgeons towards the use of standard endovascular repair (EVAR) for infrarenal abdominal aortic aneurysm (AAA) outside the instruction for use (IFU) through a Delphi consensus endorsed by the Italian Society of Vascular and Endovascular Surgery (Societa Italiana di Chirurgia Vascolare ed Endovascolare - SICVE). METHODS: A questionnaire consisting of 26 statements was developed, validated by an 18 -member Advisory Board, and then sent to 600 Italian vascular surgeons. The Delphi process was structured in three subsequent rounds which took place between April and June 2023. In the first two rounds, respondents could indicate one of the following five degrees of agreement: 1) strongly agree; 2) partially agree; 3) neither agree nor disagree; 4) partially disagree; 5) strongly disagree; while in the third round only three different choices were proposed: 1) agree; 2) neither agree nor disagree; 3) disagree. We considered the consensus reached when &gt;70% of respondents agreed on one of the options. After the conclusion of each round, a report describing the percentage distribution of the answers was sent to all the participants. RESULTS: Two -hundred -forty-four (40.6%) Italian Vascular Surgeons agreed to participate the first round of the Delphi Consensus; the second and the third rounds of the Delphi collected 230 responders (94.3% of the first -round responders). Four statements (15.4%) reached a consensus in the first rounds. Among the 22 remaining statements, one more consensus (3.8%) was achieved in the second round. Finally, seven more statements (26.9%) reached a consensus in the simplified last round. Globally, a consensus was reached for almost half of the proposed statements (46.1%). CONCLUSIONS: The relatively low consensus rate obtained in this Delphi seems to confirm the discrepancy between Guideline recommendations and daily clinical practice. The data collected could represent the source for a possible guidelines' revision and the proposal of specific Good Practice Points in all those aspects with only little evidence available