This study investigated the effectiveness of atmospheric cold plasma (ACP) treatments at different voltages (10-50 kV) in mitigating muscle protein degradation and maintaining the quality of Pacific white shrimp (Litopenaeus vannamei) during refrigerated storage at 4 °C. Shrimp samples were treated with ACP at various voltages, and physicochemical indices, including trichloroacetic acid (TCA)-soluble peptides, acid-soluble nitrogen (ASN), and free amino acid (FAA) contents, were monitored throughout storage. Proteomic analysis was conducted to identify differentially expressed proteins (DAPs), followed by Gene Ontology (GO), KEGG pathway, and protein-protein interaction analyses to elucidate functional alterations. Findings indicated that the 50 kV ACP treatment markedly retarded the buildup of TCA-soluble peptides and ASN, and alleviated fluctuations in FAA levels relative to the control. Over the 0-12-day storage interval, fewer differentially abundant proteins (DAPs) were identified in the 50 kV-treated group than in the untreated control group. Bioinformatic analyses suggested that these DAPs were largely linked to the preservation of protein structure. Correlation analysis suggested that reactive species generated by ACP induced oxidative modification of key endogenous proteases (cathepsins B, L, D, and calpains), leading to conformational changes that inhibited enzyme activity. Consequently, hydrolysis of myosin heavy chains and actin was reduced, preserving myofibrillar integrity, protein solubility, and cytoskeletal stability. Overall, ACP treatment effectively impeded protein degradation through a non-thermal mechanism, offering a sustainable preservation strategy to extend the shelf life of shrimp and other high-protein aquatic products while minimizing quality deterioration and economic losses.

Disseminated histoplasmosis is a leading AIDS-related opportunistic mycosis and carries high mortality risk. A simplified induction strategy using single high-dose of liposomal amphotericin B (L-AmB) has shown promise for disseminated disease, but persons with central nervous system (CNS) involvement were excluded from the clinical trial. We describe a 35-year-old man with advanced HIV disease (CD4 24 cells/µL) and a high fungal burden who was diagnosed with disseminated histoplasmosis and, subsequently, culture-confirmed CNS involvement. He was administratively withdrawn from the trial when CNS involvement was noted. He was treated within a randomized trial with a single 10 mg/kg dose of L-AmB, followed by antiretroviral therapy (ART) and itraconazole consolidation therapy, with full clinical recovery, normalization of cerebrospinal fluid biochemistry findings, and microbiological clearance. This suggests that a single high-dose L-AmB induction strategy, combined with effective ART and azole consolidation therapy, may be viable in selected patients with CNS involvement.

The development of craniofacial bones is highly dependent on the intramembranous ossification process driven by orofacial mesenchymal stem cells (OMSCs). Optineurin (OPTN), a traditional mitophagy receptor, promotes OMSC matrix mineralization by mediating the extracellular release of amorphous calcium phosphate (ACP) through the initiation of mitophagy via liquid-liquid phase separation. However, the dynamic mechanisms by which OPTN regulates mitochondrial engulfment remain elusive. In this work, we show that in both zebrafish and mouse models, Optn absence causes systemic bone loss and craniofacial abnormalities. Mechanistically, OPTN recruits thyroid receptor-interacting protein 6 (TRIP6) to the surface of damaged mitochondria, inducing local F-actin polymerization and remodeling, which relies on cytoskeletal mechanical forces to support the bending and closure of the phagophore. The depletion of OPTN or TRIP6 significantly impairs mitophagy flux, triggering a massive accumulation of dysfunctional mitochondria. Overall, this study elucidates that the OPTN-TRIP6-F-actin axis ensures autophagosome closure through cytoskeletal remodeling, revealing a novel paradigm in which cytoskeletal mechanical regulation maintains mitochondrial homeostasis to orchestrate craniofacial osteogenesis.

BackgroundAdvance Care Planning (ACP) enables people with progressive diseases to document and communicate future treatment preferences. For Parkinson's disease (PD) and atypical parkinsonism (AP), ACP is particularly important because of disease complexity, symptom burden, the progressive nature of the disease, and cognitive decline, yet its implementation across care settings remains unclear.MethodsWe conducted a medical chart review of 189 deceased patients with PD or AP who died in nursing homes (n = 91), hospitals (n = 58), or at home (n = 40) in the Netherlands. For patients who died at home, data were obtained from their general practice records. Data on treatment preferences, care discussions and palliative care goals were extracted at one year and six weeks before death.ResultsDocumentation of treatment preferences increased towards death but differed markedly between settings. Nursing homes recorded resuscitation and ICU limitations early and comprehensively, whereas hospitals made major updates only in the final weeks of life. In general practices, ACP documentation was limited. Nursing homes and hospitals more frequently addressed disease-specific scenarios, palliative sedation and end-of-life issues, while general practices more often reported preferred place of death and euthanasia. Palliative care goals in nursing homes covered all domains, while in hospitals and general practices they mainly related to physical aspects.ConclusionACP remains inconsistently documented and frequently delayed until the final weeks of life, when crises or cognitive decline may hinder meaningful participation. Earlier, iterative ACP-supported by clinician training, PD-specific guidance, and system-level measures such as ACP prompts in electronic medical records-could foster a more proactive and person-centered palliative approach aligned with patient values throughout the disease trajectory.

Improving the aqueous solubility of moderately soluble drugs using safe and sustainable solvent systems remains an important challenge in pharmaceutical formulation. In this study, a series of aqueous natural deep eutectic solvents (NADESs) were systematically designed and evaluated to enhance the solubility, stability, and biocompatibility of acetaminophen (ACP). The effects of hydrogen bond donor type, hydrogen bond acceptor type, molar ratio, and NADES concentration were investigated to identify the optimal formulation. Among the tested systems, the aqueous betaine chloride-acetic acid NADES (BeCl-Ace) at a molar ratio of 1 : 4 and a concentration of 70% exhibited the highest solubilization capacity, increasing ACP solubility by 25.28 and 22.14-fold at 25 and 37 °C, respectively, compared with water. Computational analyses based on electrostatic potential mapping, binding energy calculations, and interaction region indicator analysis revealed that the enhanced solubility originates from cooperative hydrogen bonding, electrostatic attraction, and van der Waals interactions between ACP and the BeCl-Ace components, particularly BeCl. In addition, the selected NADES formulation significantly improved ACP stability under thermal stress, retaining 89.13% of the initial drug content after 30 days at 37 °C, compared with 70.20% in water. Cytotoxicity evaluation on HEK-293A cells further confirmed good biocompatibility, with cell viability remaining above 92% across the tested concentration range. These findings indicate that aqueous BeCl-Ace NADESs represent a promising and sustainable solvent system for improving ACP formulation performance.

In this work, we present a detailed mechanistic and solvent-effect study of the multicomponent synthesis of 2-amino-3-cyano-4H-pyran (ACP) derivatives catalyzed by a water-soluble imidazolium-functionalized polymer (PEI.IL)(?)a synthetic enzyme (synzyme). Rather than focusing solely on synthetic scope, this study dissects how solvent properties and reaction equilibria govern reaction productivity and pathway selection in this multicomponent transformation. Solvent effects were quantitatively analyzed using the Kamlet-Abboud-Taft model, revealing that hydrogen-bond acidity (α), hydrogen-bond basicity (β), and dipolarity/polarizability (π*) play distinct and competing roles at different stages of the reaction. Control experiments, combined with systematic variation of reaction conditions, demonstrate that product formation proceeds predominantly through a pathway initiated by the reversible condensation between benzaldehyde and malononitrile, followed by enol-mediated a new C-C bond formation. Reaction productivity was treated as a thermodynamically oriented descriptor, allowing correlations between solvent effects, keto-enol equilibria, and overall efficiency to be established. The methodology affords a series of ACP derivatives in good to excellent yields under optimized conditions, with the catalyst remaining recyclable over multiple cycles without loss of activity. By integrating solvent-effect analysis, equilibrium considerations, and mechanistic control experiments, this work provides an experimentally grounded framework for understanding and optimizing multicomponent reactions beyond conventional yield-based approaches.

Long COVID is a multi-system, heterogeneous condition that manifests as new or worsened symptoms lasting at least 3 months following acute SARS-CoV-2 infection, often despite normal results on routine diagnostic testing. Common manifestations include fatigue, post-exertional malaise, cognitive impairment, and orthostatic intolerance. Primary care clinicians are uniquely positioned to identify and manage Long COVID due to their longitudinal relationships with patients and holistic understanding of their patients' health. However, diagnostic uncertainty, time constraints, and limited access to specialty care present significant challenges, and sometimes hesitation, to addressing Long COVID in the primary care setting. This focused narrative review provides a pragmatic, primary care-focused approach for the evaluation and management of Long COVID. Using patient-centered vignettes, we first describe common clinical presentations. Next, we provide a framework for the pathophysiological mechanisms suspected to contribute to persistent symptoms after SARS-CoV-2 infection. We then present a simple approach to Long COVID diagnosis and management that leverages primary care clinicians' skillset and highlights unique aspects of Long COVID care (e.g., diagnostic workup to evaluate for competing or coexisting diagnoses; management strategies focused on validation and supportive care, including activity pacing, multidisciplinary rehabilitation, and consideration of symptom-based medication trials). We offer practical strategies to effectively, yet efficiently, address a possible Long COVID diagnosis within the context of brief primary care visits (e.g., prioritize the most bothersome or addressable symptom(s) first, focus on one or two symptoms each visit, leverage longitudinal follow-up). We conclude with evidence-based strategies for prevention counseling, including infection prevention and early outpatient treatment during acute infection. While we remain optimistic that reliable diagnostic biomarkers and curative treatments will one day be available, until then, this primer supports primary care clinicians in recognizing Long COVID and supporting their patients who are living with it, despite the constraints of our current healthcare environment.

Adamantinomatous craniopharyngiomas (aCPs) are complex intracranial neoplasms that arise in the sellar and suprasellar region of the brain, affecting the endocrine and nervous systems, causing severe sequelae. Although most of aCPs are driven by somatic mutations in β-catenin (CTNNB1), some cases remain genetically unresolved. Here, we show that disruption of the Wnt-antagonist and tumour suppressor adenomatous polyposis coli (Apc) leads to pituitary tumours that present all classic molecular and histopathological hallmarks of aCPs. Mechanistically, we show that a hypomorphic allele of Apc leads to tumour development and that bi-allelic loss of Apc in the Sox2+ve pituitary stem cells can initiate aCP formation. Apc-driven tumour-initiating cells show a distinct transcriptional signature of p21-mediated senescence-associated secretory phenotype (SASP) with methylomic analyses revealing shared epigenetic alteration of SASP-like inflammatory programme across aCP subtypes. Our data demonstrate a causal role of disruption of Apc in driving aCPs, reinforcing the importance of genetic testing for mutations in APC in patients with aCP. Furthermore, our work highlights the need to screen patients with familial adenomatous polyposis (FAP) or the spectrum of APC-pathogenic syndromes for aCP development early in life.

BackgroundNative American (NA) patients experience persistent inequities in advance care planning (ACP) and often present to tertiary centers with critical illness; culturally respectful serious-illness communication is therefore essential in emergency and intensive care settings.InterventionWe developed and implemented a nursing-led, multidisciplinary educational initiative at a large academic medical center serving patients from >50 tribal nations. Informed by a literature review, local practice needs, and expert input from a Native American Patient Experience Ambassador, we created (1) an "Honoring Your Choices" (HYC) communication card with culturally sensitive prompts for serious illness conversations and (2) a quick reference guide (QRG) outlining key cultural considerations and unit-specific resources. Tools were disseminated to seven intensive care units and the emergency department through nurse leader engagement, unit rounding, and staff education, and the initiative was presented at departmental Grand Rounds/didactics with pre/post surveys to assess impact.ResultsTwenty-seven pre-session and twenty-four post-session surveys were collected from clinicians and trainees. Prior to the session, >80% of respondents reported being not or only slightly confident discussing end-of-life issues with NA patients/families; post-session surveys demonstrated increased self-reported confidence in recognizing when cultural beliefs may influence care and in conducting end-of-life discussions.ConclusionsThis initiative demonstrates how culturally congruent approaches to serious illness conversations may support more meaningful engagement with NA patients and families. Our experience suggests that developing and evaluating a brief, point-of-care communication tool like Honoring Your Choices may be accomplished with existing interdisciplinary partnerships, offering a practical model that other institutions can readily mirror.