Single-nucleus RNA sequencing (snRNA-Seq) has transformed our understanding of complex tissues, providing insights into cellular composition and heterogeneity in gene expression between cells, and their alterations in development and disease. High costs however constrain the number of samples analysed. Sample pooling and their demultiplexing following sequencing based on prior labelling with antibodies or lipid anchors conjugated to DNA barcodes (cell hashing and MULTI-seq), or using genetic differences between samples, provides a solution. However, there remains no comprehensive evaluation of these demultiplexing tools to guide selection between them. Here, we benchmark the leading software (Vireo, Souporcell, Freemuxlet, scSplit) used for sample demultiplexing using genetic variants. We further compared obtaining genetic variants from SNP array analysis of gDNA and from sample-matched bulk-RNA-Seq data, identified using three different variant calling tools (BCFtools, cellSNP, FreeBayes). Demultiplexing performance was evaluated on simulated multiplexed datasets comprising two, four, and six samples with doublet percentages between 0% and 30%, and validated against demultiplexing using sex-linked genes. Software implementation and execution were evaluated by run speed, robustness, scalability, and usability. Our results show that all tools excluding scSplit provide high recall and precision with an accuracy of 80%-85%. Vireo achieved the best accuracy. Demultiplexing tools were differentially affected by the variant calling tool with which it was paired. For all tools, accuracy decreased with the increasing percentage of doublets. Deployment of demultiplexing during analysis of pooled real-world 10x RNA-Seq data from the human heart and from different species is shown, as are advantages for doublet detection and removal.

Klotho has been implicated in blood pressure regulation through its effects on vascular integrity, calcium-phosphate homeostasis, and oxidative stress pathways. However, population-level data exploring its association with hypertension remain inconclusive.

Data from the U.S. National Health and Nutrition Examination Survey (NHANES; 2007-2016) were analyzed using weighted multivariate generalized linear regression models. Klotho levels were evaluated as both continuous and categorical (quartiles) variables. A nonlinear dose-response relationship was further explored using spline regression. To assess potential causality, a two-sample Mendelian randomization (MR) analysis was conducted using the inverse-variance weighted (IVW) method. External validation was performed in an independent Asian cohort.

In NHANES, no statistically significant association was observed between serum Klotho and hypertension, either as a continuous variable or by quartiles after adjusting for coronary heart disease (CHD) or hyperlipidemia (e.g. Q4: OR = 0.89 [95% CI: 0.79-1.00], p = 0.06 for CHD adjustment). However, nonlinear analysis revealed a potential inflection point at 1342.8 pg/mL, suggesting a threshold-dependent relationship. The MR analysis yielded no evidence of a causal association between genetically predicted Klotho levels and hypertension risk (IVW OR = 1.005 [95% CI: 0.999-1.011], p = 0.09). Findings from the small Asian cohort were consistent with null associations (OR = 0.999 [95% CI: 0.991-1.007], p = 0.80).

Across three distinct populations, no robust linear or causal relationship between Klotho levels and hypertension was identified. However, due to the limited sample size of the Asian cohort, future studies are warranted to validate these findings in larger populations.

Aneurysmal subarachnoid hemorrhage (aSAH) with World Federation of Neurological Societies (WFNS) grade V has a high mortality rate and poor prognosis. Some patients with WFNS grade V aSAH have had good outcomes after aggressive treatment; however, outcome predictions based on routine examinations and findings obtained at admission are yet to be reported. This study aimed to develop a decision tree model for predicting outcomes of patients with WFNS grade V aSAH to aid decision-making for treatment strategy.

A multicenter study with retrospective and prospective data collected from 201 (derivation cohort) and 26 (validation cohort) patients with WFNS grade V aSAH, respectively, was conducted. Clinical outcomes were divided into good (Modified Rankin Scale [mRS] score at the time of discharge: 0-2) and poor (mRS score: 3-6) outcomes. A decision tree model was developed for the derivation cohort using the classification and regression tree method with clinical data including laboratory findings; it was named OPAS-V (Outcome Prediction in Aneurysmal Subarachnoid hemorrhage with WFNS grade V). The performance of the model was evaluated by area under the curve (AUC) and overall accuracy in both cohorts.

OPAS-V comprised 3 metrics; the percentage of lymphocytes (< 49.9% or not), age (> 50 yrs or not), and glucose to potassium ratio (≥ 3.2 or not). The model achieved an AUC of 0.828 (95% confidence interval: 0.712-0.944) and overall accuracy of 0.930. Moreover, the model performed well in the validation cohort with an AUC of 0.727 (95% confidence interval: 0.441-1) and overall accuracy of 0.885.

This study developed the first decision tree model for predicting outcomes of patients with WFNS grade V aSAH, based on simple findings obtained at admission. This may aid clinicians in determining treatment strategies for severe conditions such as WFNS grade V aSAH.

To compare the efficacy and safety of ticagrelor versus clopidogrel in stroke patients who were CYP2C19 loss-of-function (LOF) carriers stratified by age and renal function.

Patients in the CHANCE-2 trial were randomized to ticagrelor-aspirin or clopidogrel-aspirin treatment. The primary efficacy outcome was occurrence of a new stroke within 90 days, while bleeding was assessed for safety. Patients were categorized based on age and estimated glomerular filtration rate (eGFR).

In patients with eGFR >90 mL/min/1.73 m², ticagrelor-aspirin was associated with a significantly lower risk of the subsequent stroke within 90 days compared with the clopidogrel-aspirin in those aged over 65 years (HR 0.53, 95% CI 0.33-0.85, p = 0.008) and under 65 years (HR, 0.67, 95% CI, 0.47-0.96, p = 0.03). While in those with eGFR 60-89 mL/min/1.73 m², ticagrelor did not show superiority over clopidogrel in reducing stroke regardless of age category (age ≥ 65: HR 1.14, 95% CI 0.71-1.84, p = 0.59; age < 65: HR 0.40, 95% CI 0.12-1.33, p = 0.13). The incidence of mild bleeding events was higher with ticagrelor-aspirin treatment in those aged < 65 years with eGFR ≥90 mL/min/1.73 m² (HR 3.33, 95% CI 2.18-5.10, p < 0.001) and in those aged ≥ 65 years with eGFR <60mL/min/1.73 m² (HR 8.68, 95% CI 1.06-71.1, p = 0.04).

Elderly patients with normal renal function appear to benefit from ticagrelor compared with clopidogrel. Both younger patients with normal renal function and those with advanced age and renal insufficiency are prone to mild bleeding.

Behcet's syndrome (BS) is a multisystem syndrome that typically manifests as recurrent oral and genital ulcers, as well as other systemic manifestations. Few studies describing the characteristics of BS among Israeli patients have been published.

To describe the characteristics of BS patients and to compare Jewish and Arab subpopulations.

We retrospectively reviewed electronic medical records and extracted demographic, clinical, laboratory, and medication data for each patient. We compared the Jewish and Arabic BS patients.

The cohort included 98 patients. Males constituted 49 (50%); mean age at the time of diagnosis was 29.9 years; 71 (72.4%) were Arab and 27 (27.6%) were Jewish. Oral ulcers were evident in 93 patients (94.9%) and genital ulcers in 54 (55.1%). Involvement of the skin, joints, eyes, gastrointestinal tract, and neurologic and vascular systems were demonstrated among 42 (42.9%), 57 (58.2%), 47 (48.0%), 8 (8.2%), 10 (10.2%), and 15 (15.3%), respectively. HLA B51 was positive in 24 of 37 (64.9%). Pathergy was positive in 8 of 12 (66.7%). Colchicine was used in 82 (83.7%), azathioprine 47 (48%), methotrexate 16 (16.3%), apremilast 10 (10.2%), cyclosporine-A 8 (8.2%), adalimumab 26 (26.5%), infliximab 12 (12.2%), cyclophosphamide 1 (1.0%), tocilizumab 2 (2.0%), and anti-coagulation 6 (6.1%). The Arab and Jewish subpopulations were significantly different regarding male proportion, 40 (56.3%) vs. 9 (33.3%), P = 0.042.

BS is more common among Arabs in northern Israel, but no significant clinical or demographic differences were found except for a higher proportion of male patients among Arabs.

Early assessment of cardiovascular disease risk plays an important role in preventing cardiovascular disease, vascular age (VA) is an important indicator for early screening of cardiovascular disease risk. This study presents a pulse signal-based dataset for VA prediction. The dataset comprises 226 subjects with 1364 pulse cycles, spanning both sexes (49.6% male, 50.4% female) and an age range of 20 to 69 years. Pulse signals were denoised by Savitzky-Golay filters, and 4th-order derivatives were calculated to extract the features of pulse signal. We applied the classic statistical model Klemera Doubal method (KDM) and five artificial intelligence models to predict VA. The experimental results showed that these models can predict VA with high accuracy and stability. It indicates that using pulse signals to predict VA is a simple, non-invasive, and effective method for assessing vascular health.

This meta-analysis aims to compare the clinical outcomes of the no-touch (NT) and conventional (CON) vein harvesting techniques in patients undergoing coronary artery bypass grafting (CABG).

We conducted a systematic review and meta-analysis following the guidelines of the Cochrane Handbook and PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) statement. We searched PubMed, Embase and Web of Science for randomised controlled trials (RCTs) comparing NT and CON vein harvesting techniques in CABG patients. Data were extracted on primary outcomes (graft failure incidence per patient, graft occlusion incidence per patient and leg infection) and secondary outcomes (revascularisation, all-cause death, myocardial infarction). Statistical analysis was performed using Review Manager V.5.4, with risk ratios (RRs) calculated for binary outcomes.

Seven RCTs involving 4176 patients were included. The NT group showed a significantly lower risk of graft failure incidence per patient (RR=0.74, p=0.0001) and graft occlusion incidence per patient (RR=0.62, p=0.0002) compared with the CON group. However, the NT group had a higher risk of leg infection (RR=1.91, p<0.00001). No significant differences were observed between groups for revascularisation (p=0.46), all-cause death (p=0.87), or myocardial infarction (p=0.95).

The no-touch vein harvesting technique is associated with reduced graft failure incidence per patient and graft occlusion incidence per patient compared with conventional harvesting techniques in CABG, though it increases the risk of leg infection. These findings suggest NT as a preferable technique for improving graft patency but highlight the need for caution regarding leg infection.

CRD42025646500.

A man in his mid-40s presented with recurrent episodes of syncope, some of which were triggered by neck extension. In-hospital telemetry documented sinoatrial arrest with a prolonged asystole of 11 s. Computed tomography of the neck demonstrated bilateral elongation of the styloid processes consistent with Eagle syndrome (ES). Vagal reflex stimulation was suspected as the underlying mechanism. A dual-chamber pacemaker was implanted prior to surgical styloidectomy. At 3-month follow-up, pacemaker interrogation revealed 2% pacing activity, with no further syncopal episodes. This case highlights an unusual cardioinhibitory presentation of ES and supports early pacing when surgical resolution of vagal stimulation remains uncertain.

Excess aldosterone production contributes to the development of hypertension and results in fibrosis with dysfunction of the heart, vasculature and kidneys. Consequently, new agents have been developed to reduce endogenous aldosterone synthesis. The primary objective of this systematic review is to describe the BP-lowering effects of aldosterone synthase inhibitors (ASIs) in hypertensive patients and, secondly, to describe their potential renal protective effects and possible influence on cortisol production and plasma potassium. We searched PubMed, Embase and ClinicalTrials.gov and included randomized controlled and clinical trials according to PICO using the review tool Covidence. Thirteen studies were included and all demonstrated BP reduction through ASI treatment. Among patients with apparent resistant hypertension, the placebo-corrected reductions in seated systolic BP were 11.0 mmHg for baxdrostat and 9.6 mmHg for lorundrostat. A significant suppression of cortisol production was found for LCI699 (osilodrostat) but not for baxdrostat, lorundrostat, BI 690517 (vicadrostat) or dexfadrostat. Studies on BI 690517 showed a reduction in urine-albumin-creatinine ratio, indicating renal protection. ASIs may increase potassium levels. We conclude that ASIs have promising BP-lowering effects with very limited effects on cortisol production and offer reno-protective effects in chronic kidney disease. Studies on hypertensive target organ damage and cardiovascular outcomes are, however, lacking.

To analyse anatomic factors of the distal landing zone (dLZ) associated with the durability of endovascular repair of thoracic aortic aneurysm (TAA).

Consecutive patients undergoing thoracic endovascular aortic repair (TEVAR) for undissected TAA were queried from a single centre from 2004 to 2022. Patient and operative factors were considered as well as detailed anatomic factors at the dLZ assessed by 3D reconstruction of pre-TEVAR imaging. The outcome of interest was the long-term risk of TEVAR failure at the dLZ.

A total of 101 patients undergoing TEVAR repair of TAA were considered, of whom 17 suffered distal TEVAR failure over a median follow-up period of 2.7 years. Two anatomic factors showed outsized influence on long-term outcomes: dLZ diameter and dLZ length (the length of non-dilated aortic tissue above the coeliac artery). Patients who progressed to distal TEVAR failure had larger dLZ diameter (34.2 mm vs 30.7 mm, P = 0.034) and far shorter dLZ length (3.8 cm vs 7.5 cm, P = 0.008). Patients with dLZ diameter greater than 35 mm had much greater risk of mortality or distal TEVAR failure within 2 years (34% vs 5%, P = 0.012), as did those with dLZ length less than 4 cm (27% vs 6%, P = 0.006).

In this study, mild dilation at the dLZ beyond 35 mm and short length at the dLZ less than 4 cm are both clear anatomic risk factors for poor long-term outcome after supraceliac TEVAR. It may be appropriate to consider repair via branched endografts landing beyond the coeliac artery in patients with these risk factors.