Browse Articles
Discover research articles across all indexed journals
A cautionary note on the potential pitfalls of using N-terminal truncated CD63 to label small extracellular vesicles
Abstract Small extracellular vesicles (sEV) are nanosized vesicles that facilitate intracellular communication. A significant research obstacle is the isolation of sEV devoid of non-sEV contaminants. Immunoaffinity capture with sEV-specific antibodies is an attractive approach to purifying sEV, but it risks disrupting the vesicles during antibody dissociation. Furthermore, immunoaffinity capture may require the modification of EV-specific proteins for the incorporation of tags on the EV surface, with unknown implications on EV production and function. The aim of this study was to investigate whether a previously reported CD63 truncation is efficient for the incorporation of small tags on the extravesicular surface. We therefore conjugated ALFA-tag to N-terminal-truncated CD63, and included nanoluciferase at the C-terminus, for luminescent tracing of the sEV. Full-length CD63-nanoluciferase was used as a control. Plasmid constructs expressing these proteins were transfected into HEK293 cells. In contrast to a previous report, the N-terminal truncation of CD63 impaired its membrane localisation and reduced the yield of EVs. Further investigation revealed that some of the tagged CD63 was co-localized with aggresomes and was preferentially secreted from the cells as soluble protein rather than being associated with sEV. These results demonstrate that CD63 truncation can impair its function and EV yield, potentially generating misleading results.
FDA approves new non-opioid pain drug
Enhancing the power quality of dual rotor wind turbines using improved fuzzy space vector modulation and super twisting sliding techniques
Record of polycyclic aromatic hydrocarbons (PAHs) from prehistoric sediments and human activity in the Lubei plain of China
Abstract Understanding the sources and dynamics of past biomass burning remain a significant challenge due to variations in paleofire combustion patterns across different temporal and spatial scales. This study integrates black carbon and polycyclic aromatic hydrocarbon (PAH) records from the Lubei Plain in the Shandong Peninsula, Lower Yellow River, to reconstruct Holocene fire regimes and their relationship with climatic shifts and human activities over the past 5000 years. During the mid-to-late Holocene (5000–3000 year BP. (calendar years before 1950)), a biomass burning levels were generally low, with a pronounced peak in low-molecular-weight PAHs (3-ring PAHs) and charcoal fluxes between 5000 and 4500 year BP, indicating increased fire activity likely driven by a short-term cold-dry event around 5000 year BP. From 3500 to 1000 year BP, three distinct episodes of low-temperature smoldering fires are identified, coinciding with deforestation and persistent droughts during the Shang Dynasty (3600–3046 year BP), the Qin and Western Han Dynasties (2200–2000 year BP), and the Sui and Tang Dynasties (1400–1100 year BP). In contrast, high-temperature flaming fires are associated with periods of intensified warfare and social upheaval, compounded by cold, arid climates during the Warring States period (2500–2400 year BP), the Eastern Han Dynasty (2000–1800 year BP), and the Wei, Jin, and Southern-Northern Dynasties (1800–1400 year BP). Over the past millennium, anthropogenic biomass burning remained elevated, reflecting sustained human influence on fire regimes. Meanwhile, Pollen and n-alkane records reveal a transition from primary forests to secondary shrubland during the late Holocene, driven by declining seasonal precipitation linked to a weakening East Asian monsoon and increased anthropogenic burning. Principal component analysis indicates that long-term deforestation primarily drove low-temperature smoldering fires, whereas high-temperature fires were more closely linked to periods of conflict. Seasonal precipitation variability, regulated by monsoonal dynamics, emerged as a fundamental control on fire regimes. This integrated analysis of PAHs, black carbon, and charcoal, coupled with multivariate statistical approaches, offers a robust framework for reconstructing fire-climate-human interactions in East Asia. The findings provide new insights into the mechanisms driving fire regimes and their long-term ecological and societal impacts.
Revisiting risk factors and incidence of postoperative tachyarrhythmias in pediatric cardiac surgery
J&J buys Intra-Cellular Therapies for US$14.6 billion, expanding its neuroscience portfolio
Robust self supervised symmetric nonnegative matrix factorization to the graph clustering
Base editing reduces prion protein
Scalable production and purification of engineered ARRDC1-mediated microvesicles in a HEK293 suspension cell system
Abstract Engineering of human ARRDC1-mediated microvesicles (ARMMs) as non-viral vehicles for delivery of gene therapies bears the potential to enable novel therapeutic paradigms. We evaluated two scalable strategies to generate ARMMs loaded with protein cargo, by transient transfection or stable cell line-based production. The upstream ARMMs production processes utilized a suspension-adapted HEK293-derived line, termed 5B8. 5B8 cells yielded robust production of ARMMs after transient transfection with the ARMMs loading construct or using a stable cell line containing a transgene that encodes the ARMMs loading cassette, in shake flasks or a stirred tank bioreactor, respectively. ARMMs were purified by ultracentrifugation (small scale) or a combination of TFF and AEX (scalable production). Both purification methods produced comparable ARMMs, in terms of size and payload incorporation. Single particle analysis showed approximately 50% were payload-containing ARMMs. Additionally, an in vivo study was conducted in mice to investigate the half-life and biodistribution of ARMMs administered intravenously. ARMMs showed rapid biodistribution predominantly to the spleen and liver and, to a lesser extent, kidneys, and lungs. The half-life of ARMMs in plasma was 6 ± 0.4 min. Altogether, this work advances knowledge on scale-up of engineered cell-derived vesicles for future in vivo delivery of therapeutic molecules.
A novel Kayvirus species phage RuSa1 removes biofilm and lyses multiple clinical strains of methicillin resistant Staphylococcus aureus
Provision, cough efficacy and treatment satisfaction of mechanical insufflation-exsufflation in a large multicenter cohort of patients with amyotrophic lateral sclerosis
Abstract In patients with amyotrophic lateral sclerosis (ALS), mechanical insufflation-exsufflation (MI-E) addresses cough deficiency to achieve major therapeutic goals: improving costal muscle and joint function, reducing atelectasis through insufflation, and clearing bronchial secretions via exsufflation. Despite its perceived benefits, there is limited systematic research on MI-E provision, symptom alleviation, or patient satisfaction. The research platform Ambulanzpartner coordinated this longitudinal observational study conducted in 12 German ALS centers from July 2018 to September 2023. Patients were enrolled based on ALS-related cough deficiency requiring MI-E therapy. The study recorded provision, reasons for withholding MI-E, clinical parameters, therapy frequency, subjective cough deficiency, and symptomatic relief. Satisfaction with MI-E therapy was determined by the likelihood of recommendation. Out of 694 ALS patients indicated for MI-E, 527 (75.9%) received the therapy. The primary reason for non-provision was that the patient had died before provision (n = 66 of 167; 39.5%). These patients were significantly more affected as represented by higher progression rates and lower cough peak flows (CPF) at the time of MI-E indication ( p < 0.05). Most patients who received MI-E used it daily (n = 290 of 370; 78.4%). Self-assessed cough deficiency correlated with clinical measurements, especially for patients with higher deficits. At follow-up visits, patients reported reduced cough deficiency ( p < 0.001). Frequent MI-E use was linked to greater symptom relief and higher likelihood of recommending the therapy. This study highlights the symptomatic and palliative potential of MI-E therapy for ALS patients.
Profiling of RNA N6-Methyladenosine methylation reveals the critical role of m6A in betaine alleviating hepatic steatosis
A covalent chemical probe for Chikungunya nsP2 cysteine protease with antialphaviral activity and proteome-wide selectivity
Abstract Chikungunya is a mosquito-borne viral disease that causes fever and severe joint pain for which there is no direct acting drug treatments. Vinyl sulfone SGC-NSP2PRO-1 (3) was identified as a potent inhibitor of the nsP2 cysteine protease (nsP2pro) that reduced viral titer against infectious isolates of Chikungunya and other alphaviruses. The covalent warhead in 3 captured the active site C478 and inactivated nsP2pro with a k inact/K i ratio of 5950 M–1 s–1. The vinyl sulfone 3 was inactive across a panel of 23 other cysteine proteases and demonstrated remarkable proteome-wide selectivity by two chemoproteomic methods. A negative control analog SGC-NSP2PRO-1N (4) retained the isoxazole core and covalent warhead but demonstrated > 100-fold decrease in enzyme inhibition. Both 3 and 4 were stable across a wide range of pH in solution and upon prolonged storage as solids. Vinyl sulfone 3 and its negative control 4 will find utility as high-quality chemical probes to study the role of the nsP2pro in cellular studies of alphaviral replication and virulence.
Association between FT4/FT3 ratio and microalbuminuria in euthyroid patients with type 2 diabetes mellitus
Lead free perovskite integrated metal organic framework as heterogeneous catalyst for efficient three component click reaction
Constructing a knowledge graph-driven intelligent data-enabled design system for mold using deep semantic understanding and intelligent decision support
Stability analysis of carbon emission trading mechanism in China based on a tripartite evolutionary game
[18F]FDG-PET and [18F]MPPF-PET are brain biomarkers for the creatine transporter Slc6a8 loss of function mutation
Abstract Pathogenic variants in the creatine transporter gene SLC6A8, reported to represent 2% of all intellectual disabilities in males, result in a spectrum of behavioral abnormalities including developmental delay, intellectual disability, and deficit in speech. While at present there are no effective treatments available, preclinical development and testing of gene therapy and other approaches to increase brain creatine are being actively pursued. In studying a mouse model of the disorder, [18F]fluorodeoxyglucose ([18F]FDG)-based positron emission tomography (PET)/computed tomography (CT) was performed to assess brain glucose metabolism in wild type and creatine transporter mutant mice (Slc6a8 -/y ). The findings demonstrate marked differences in glucose metabolism in the brains of wild type and Slc6a8 -/y mice. In conducting behavioral phenotyping studies, notable abnormalities in behavior in the murine model led to additional studies in serotonin-mediated activity. Serotonergic signaling differences were detected between wild type and Slc6a8 -/y mice using 4-(2′-methoxyphenyl)-1-[2′-(N-2″-pyridinyl)-p-[18F]fluorobenzamido]ethylpiperazine ([18F]MPPF). These data demonstrate that [18F]FDG-PET and [18F]-MPPF-PET may serve as appropriate and sensitive biomarkers that could be used to assess the efficacy of not only new approaches in treating mutations of the creatine transporter SLC6A8 and their effectiveness in normalizing brain metabolism but also in enhancing our understanding of the mechanism of brain dysfunction that occurs in this complex brain disorder.