The capacity to adapt is essential for a population to avoid extinction in a changing world and is recognized as a global conservation priority. Adaptation requires additive (heritable) genetic variation for traits that influence survival and fecundity, but measuring this variation is difficult, particularly in species of conservation concern. Instead, molecular genetic diversity is often used to infer adaptive potential. However, previous research has cast doubt on the suitability of traditional molecular markers (allozymes and microsatellites) for this purpose given their weak relationship with heritability-a common measure of additive genetic variance. Advances in sequencing technology have since shifted focus toward nucleotide diversity and variation in functional regions, but their practicality for predicting adaptive potential remains debated. Furthermore, heritability itself is a poor proxy for adaptive potential because it depends on environmental variance. We collated 2,113 published estimates of evolvability-a measure of additive genetic variance that avoids environmental confounding-across 193 eukaryotic species, and evaluated how well evolvability is predicted by molecular diversity. We find that microsatellite and nucleotide diversity are not significantly correlated to each other, and neither predict evolvability. Nucleotide diversity explains 1.1% of interspecific differences in evolvability and doubling nucleotide diversity only corresponds to a 11.7% increase in evolvability. With theoretical work, we show that such weak associations are expected. Together, our results suggest that simple molecular measures of genetic variation are insufficient for predicting adaptive potential and reliance on these metrics risks misinforming conservation management.
Opioids account for 76% of drug overdose deaths in the United States, with nearly 80,000 opioid overdose deaths annually. The risk of overdose is dynamic and influenced by rapidly changing behaviors and contexts that are not well captured by retrospective or infrequent assessments. Ecological momentary assessment (EMA) allows repeated near real-time reporting of behaviors and experiences in natural settings. This study evaluated the feasibility and acceptability of smartphone-based EMA among people who use opioids to monitor opioid use, overdose experiences, and naloxone access. Participants were recruited through the New Haven Syringe Services Program in New Haven, Connecticut, and completed fixed-time, twice-daily EMA prompts for 30 days using a smartphone app. EMA measures assessed behaviors and experiences occurring within the previous 12 hours, including opioid craving, plans to use drugs, opioid use, overdose experiences, and naloxone access. Feasibility was assessed through recruitment, retention, and EMA completion rates, as well as the reporting of drug use-related behaviors. Acceptability was evaluated using exit surveys assessing ease of use, burden, and privacy, as well as qualitative interviews. Of the 13 screened individuals, 10 were enrolled and completed the baseline assessment. A total of 9 (90%) participants completed EMA and responded to 411 prompts, with an overall compliance rate of a mean of 85.0% (SD 8.7%). Compliance was similar across morning (84.2%, SD 13.1%) and evening (85.8%, SD 11.2%) prompts. Participants reported opioid cravings (274 reports), plans to use illicit drugs (252 reports), and opioid use (411 reports) during the study period. Two participants reported overdose events (5 reports) and were carrying naloxone during each overdose event. Acceptability ratings were high: all participants reported that EMA prompts were easy to understand and private, and most did not find them burdensome. Qualitative feedback further highlighted the ease of use, integration into daily routines, and increased self-awareness, although some participants reported emotional discomfort when reflecting on their substance use. Smartphone-based EMA was feasible and acceptable among people who use opioids recruited through a syringe service program. EMA may support the monitoring of opioid use, overdose experiences, and naloxone access, and it may inform future digital health interventions aimed at reducing harm, including overdose risk, among people who use opioids.
Peripheral membrane proteins (PMPs) are critical mediators of signaling cascades initiated at the cell surface. Their functions depend on their innate ability to interact dynamically with membranes in response to changing cellular conditions. This membrane recruitment may occur via high-affinity interactions with specific lipids/proteins or via transient, low-affinity interactions with the membrane. These weak and dynamic interactions, which are critical regulators of PMP function, are challenging to capture. Taking Bruton's tyrosine kinase (BTK), a nonreceptor tyrosine kinase essential for B cell activation, we demonstrate a native mass spectrometry platform to understand lipid-mediated recruitment of PMPs by directly studying it from lipid bilayers customized to target membranes. Our data demonstrate that BTK recognizes phosphatidylserine (PS) independently of phosphatidylinositol (3, 4, 5) phosphate (PIP3) binding. We show that PS-bound BTK retains PIP3 binding via high-affinity sites, while exhibiting PIP3-independent basal membrane recruitment. Biochemical assays show that this PS-mediated recruitment sensitizes BTK to PIP3-mediated activation at near-physiological PIP3 concentrations. Thus, we propose a two-step model for BTK membrane recruitment and activation. A low-affinity interaction with high-copy number PS enables plasma membrane recruitment of BTK and increases its membrane-bound concentration. Upon B cell activation, this prerecruited, membrane-bound BTK population localizes to PIP3-rich domains via electrostatic gliding along the membrane, driven by low-affinity PS and high-affinity PIP3 binding. This indicates a cooperative mechanism in which PS can amplify B cell signaling by increasing membrane-bound BTK levels. Our work demonstrates a general model of regulation of PH domain-containing proteins by weak protein-lipid interactions, which can be extended to other PMPs.
Lateral lumbar interbody fusion (LLIF) is a minimally invasive spinal fusion technique utilizing a lateral transpsoas approach to access the anterior column of the spine. Vascular injury is a rare complication but difficult to manage due to the minimally invasive nature of the procedure, making it critical to carefully review preoperative imaging prior to committing to a lateral approach. A 67-year-old female presented with progressive low back pain and left lower extremity pain and sensory changes. MRI showed a grade 1 L4-5 spondylolisthesis with left-sided neuroforaminal stenosis. The patient was recommended for LLIF; however, on further review of the MRI findings, an abnormally dilated left gonadal vein (GV) was identified along the psoas muscle. Subsequent imaging revealed compression of the left renal vein between the superior mesenteric artery and aorta, causing venous outflow obstruction. Therefore, the surgical approach was changed to avoid potential vascular complications. GV injury is a rare but major complication of lateral spine surgery. This case highlights the importance of careful preoperative imaging and anatomical review as lateral approaches become more common, and acts as a cautionary report for vascular review prior to LLIF. https://thejns.org/doi/10.3171/CASE26180.
Fractional radio frequency microneedling (FRM) is an effective nonsurgical facelift modality supported by growing clinical evidence. While generally well tolerated, postprocedural erythema, pain, swelling, and bruising typically last up to 7 days. Other serious complications include prolonged swelling, infection, persistent grid mark, burns, and hypo- or hyperpigmentation. Recombinant human collagen has demonstrated potent wound-healing and anti-inflammatory properties, which could be harnessed for post-FRM recovery. To date, there is a lack of published studies evaluating the use of topical agents to improve postprocedural recovery following FRM. This case report describes the early experience of using topical recombinant collagen post FRM for nonsurgical facelift. Two 40-year-old female patients with Fitzpatrick skin type IV underwent FRM for nonsurgical facelift. The procedure was performed with standard energy settings. Immediately post procedure, topical recombinant collagen was applied to the treated areas. Either clinical photography or cross-polarized imaging was used to objectively capture the changes in facial erythema and swelling. Both cases demonstrated a marked reduction of erythema and inflammation within 1 to 2 hours after application of topical recombinant collagen. These observations appeared earlier compared to the typical timeframe of 1 to 3 days. No adverse events were reported. Topical recombinant collagen application post FRM was associated with an early reduction of skin erythema and inflammation.
The aging of salivary glands (SGs) leads to xerostomia, impaired oral defenses, and a compromised quality of life, yet the biology of human submandibular gland (SMG) aging remains poorly understood due to limited availability of human tissues and differences in anatomy and physiology between rodent and human SGs. The common marmoset (Callithrix jacchus) is a short-lived non-human primate (NHP) with SMG histology that closely resembles humans and has the potential to provide a translational model for investigating SG aging. In this study, we examined the age-associated structural and molecular alterations that occur in marmoset SMGs and evaluated whether chronic oral rapamycin treatment, a gerotherapeutic mTOR inhibitor, mitigates these age-related changes. Histological and immunofluorescence analyses revealed that there was a substantial decline in mucous and serous acinar units, expansion of ductal and collagen-rich stromal compartments, increased extracellular matrix remodeling, elevated dysregulation of lipid metabolism, and increased numbers of apoptotic and senescent cells in the SMG with aging. Rapamycin treatment was associated with partial preservation of acinar structure and a reduction in fibrosis, lipid accumulation, and number of apoptotic and senescent cells that resulted in an overall tissue phenotype that more closely resembled that of younger animals. These findings suggest that the aging marmoset SMG recapitulates key features of human SG aging and that rapamycin treatment was associated with an attenuation in the hallmarks of SG degeneration. The results suggest that the marmoset represents a valuable NHP model for studying SG aging biology and testing therapeutic strategies aimed at attenuating age-related structural degeneration associated with SG dysfunction.
Doctors have been identified as having a crucial role in responding to anticipated and experienced stigma of People with Mental Illness (PWMI). This paper aims to evaluate the effectiveness of the READ (Responding to Experienced and Anticipated Discrimination), an anti-stigma training for medical students, by measuring changes in their knowledge, attitudes, and skills, in responding to patients anticipated and experienced discrimination. The Mental Health Knowledge Schedule (MAKS), the Mental Illness Clinicians' Attitudes version 2 (MICA2), and an OSCE (Observed Structured Clinical Examination) were used to determine participants' knowledge, attitudes, and behaviours towards PWMI before and immediately after the training. There was evidence of difference in MICA2 mean total scores in the intervention group were compared to the control group after adjusting for age, gender and MICA baseline mean total scores (MD: -7.88; p < 0.001; 95% CI: -10.23 to -3.96). Moreover, the intervention group was 4.45 times more likely to be scored "pass" in the OSCE compared to the control group (p = 0.046, 95% CI: 1.03 to 19.26) after adjusting for age, gender and OSCE baseline scores. The positive changes in students' attitudes and skills after the READ training should encourage further research on the causal pathways of this positive relationship.
Large-volume autologous fat grafting for aesthetic augmentation is often complicated by cysts and fibrosis, suggesting that its survival mechanisms differ from those of small-volume grafts. To investigate the survival biology of large-volume fat grafts and evaluate the effects of mechanical pressure on tissue structure, viability, and long-term graft outcome. A 3-dimensional in vitro model using human adipose tissue in a scaffold was established. Constructs were cultured under 0, 6 or 12 mmHg of pressure to assess structural integrity, solute permeability, cell viability and molecular changes. The viable outer layer was then transplanted into nude mice to evaluate volume retention, necrosis and fibrosis. In vitro, a porous interstitial network supported tissue survival to a depth of approximately 8 mm. Pressure dose-dependently disrupted this network, reducing solute permeability from 76.4% to 21.3% and impairing viability. Pressure also induced a form of "latent injury," consistent with YAP-related mechanotransductive signaling, lineage-related molecular changes and metabolic stress. In vivo, although gross volume retention was similar among groups, pressure-conditioned grafts developed marked necrosis, with fibrosis increasing from 22.1% in controls to 53.3% in the 12 mmHg group at 8 weeks. Large-volume fat graft survival depends in part on the integrity of a pressure-sensitive porous network. Mechanical pressure may induce a clinically relevant "latent injury," consistent with molecular changes observed in vitro, which may contribute to later fibrosis. Minimizing mechanical stress during harvest, processing, handling and implantation may help preserve graft integrity and improve long-term outcomes.
Parkinson disease (PD) is characterized by motor symptoms as well as progressive cognitive decline leading to long-term functional impairment and diminished quality of life. Mild cognitive impairment in PD (PD-MCI) is a risk factor for developing PD-related dementia. PD-MCI provides a window to assess interventions that can improve cognition. Repetitive transcranial magnetic stimulation (rTMS) shows promise as an effective treatment to improve cognitive performance. This study aims to test the safety and feasibility of a 10-session, high-frequency rTMS protocol applied to the left dorsolateral prefrontal cortex and the rTMS efficacy in improving cognitive test performance among veterans with PD-MCI. This is a double-blind randomized controlled trial. We will enroll US military veterans with PD-MCI. Participants will be randomized to either active or sham rTMS treatment groups, each with 10 treatment sessions (2 sessions/day). Treatment need not be consecutive; rather, they can be spread across approximately 10 days (eg, Monday, Wednesday, Thursday, Monday, Tuesday, and Wednesday). Participants randomized to active rTMS will receive stimulation applied to the left dorsolateral prefrontal cortex at 110% the resting motor threshold, a 15-Hz rate, 5 seconds per train of pulses, a 10-second intertrial interval, and 40 trains of pulses per session. Each patient will receive approximately 3000 pulses per session. Sham stimulation will be administered at the same parameters as real rTMS; however, no magnetic field will be produced on the placebo side of the active or placebo coil. This protocol was approved by the Edward Hines Jr Veterans Administration Hospital and Jesse Brown Department of Veterans Affairs Medical Center institutional review boards. This study includes a Food and Drug Administration investigational device exemption (G190076). Safety will be assessed using the number of research-related adverse events experienced by the active rTMS group compared to the sham rTMS group. Feasibility will be assessed using protocol completion rates. To examine preliminary effects of rTMS, participants will complete a standardized neurocognitive battery assessment at baseline, end point, and 1-month follow-up. The primary study outcome is the change in score from baseline to end point on the National Institutes of Health-sponsored Executive Abilities: Measures and Instruments for Neurobehavioral Evaluation and Research Executive Composite score. This project was funded in June 2019, with additional funding secured in April 2024. As of April 2025, a total of 18 veterans with PD-MCI have completed the randomized controlled trial phase. Data collection is ongoing and will be completed by March 2027. We expect the results of this study to be available by March 2028. The knowledge gained on the safety, feasibility, and efficacy of rTMS will set the stage for future research optimizing therapeutic gains for existing cognitive rehabilitation treatments or developing new and adjunct treatments for PD-MCI. ClinicalTrials.gov NCT03836950; https://clinicaltrials.gov/study/NCT03836950. DERR1-10.2196/77241.
Living guidelines are developed for selected topic areas with rapidly evolving evidence that drives frequent change in recommended clinical practice. Living guidelines are updated on a regular schedule by a standing expert panel that systematically reviews the health literature on a continuous basis, as described in the ASCO Guidelines Methodology Manual. ASCO Living Guidelines follow the ASCO Conflict of Interest Policy Implementation for Clinical Practice Guidelines. Living Guidelines and updates are not intended to substitute for independent professional judgment of the treating clinician and do not account for individual variation among patients. See appendix for disclaimers and other important information (Appendix I and Appendix II). Updates are published regularly and can be found on the ASCO Publications website.
This viewpoint develops the futures framework for clinical artificial intelligence governance (FF-CAIG), a conceptual and anticipatory framework for organizing emerging governance challenges in clinical artificial intelligence (AI). Although life cycle-oriented oversight is increasingly reflected in clinical AI regulation and institutional governance, existing approaches remain more developed for near-term validation, current-state assurance, and retrospective risk detection than for longer-horizon sociotechnical change. This gap is increasingly relevant, as AI systems become more complex, adaptive, and autonomous, and as they become more deeply embedded in care relationships and accountability structures. FF-CAIG is grounded in 3 futures methodologies: the 3 horizons model, scenario planning, and causal layered analysis. It is operationalized through an emerging clinical AI risk taxonomy that links these methods to governance domains. Its practical outputs include horizon classification, risk-domain mapping, scenario stress-testing findings, accountability-chain mapping, and horizon-scaled minimum governance actions for deployment or continued use. Applied across near-term, transitional, and longer-term horizons, the framework proposes cross-horizon priorities, including stronger predeployment equity evaluation, clearer life cycle accountability, clinician AI oversight competencies, and safeguards for increasingly autonomous or AI-mediated care systems. We illustrate FF-CAIG through 3 representative clinical AI deployment patterns and discuss its limitations, including differential compliance burdens, risks of overdocumentation, variable feasibility across jurisdictions, and the need for empirical validation. FF-CAIG is intended not as a prescriptive policy instrument or validated assessment tool, but as a structured analytic approach for regulators, health system leaders, developers, and researchers seeking prospective and systems-oriented approaches to clinical AI governance.
In individuals with schizophrenia receiving either clozapine or olanzapine, this study examined changes in 1) body weight and other cardiometabolic measures and microbiota biodiversity and composition between commencement and completion of 24-week semaglutide intervention; 2) body weight between commencement and 76-week follow-up. 24-week intervention (16-weeks full-dose (1.0 mg/week) after 8-weeks' titration) of open-label nurse-administered semaglutide in a public mental health setting, with one-year post-intervention follow-up (76-week trial-completion). people with schizophrenia without diabetes receiving clozapine or olanzapine with BMI > 27 kg/m2. %body weight change at 24-weeks, and 76-weeks. Secondary endpoints: %change in waist circumference, HbA1c at 24-weeks and 76-weeks, body composition at 24-weeks. Gut microbiota changes were compared at baseline, 10-weeks and 24-weeks intervention completion. Mean age: 41.5 years (range 18-61), 65.4% female. Intervention completed by 65.4% (n = 17/26). 24-week intervention: intention-to-treat body weight reduction: -9.8% (95% CI: [-12.7%, -6.8%], p < 0.001) or - 10.1 kg (95% CI [-13.6, -6.6]); waist circumference reduction: -7.3% (95% CI: [-10.1%, -4.4%], p < 0.001); HbA1c non-significant reduction: -5.3% (95% CI [-10.4%, 0.1%], p = 0.055). Microbial alpha diversity decreased as time on semaglutide increased, with enrichment of Parasutterella excrementihominis. Trial completion: 88.2% (n = 15/17). Average body weight change baseline-76-weeks: -5.1% (95% CI: [-8.3%, -1.9%], p = 0.001) or - 5.3 kg (95% CI: [-8.9, -1.7]). Semaglutide was associated with significant weight loss in overweight/obese people with schizophrenia. These benefits attenuated following semaglutide discontinuation. Gut microbial compositional differences consistent with improvement in health outcomes may occur in semaglutide-treated people living with schizophrenia.
Orexin neurons regulate physiological functions, such as energy homeostasis, wakefulness, and motivated behaviors. However, studies linking orexin neuron activity to behavior via selectively activating/inactivating these inputs in a temporally controlled manner in rats are scarce. Here, we examined the role that orexin neurons play in motivated behavior in transgenic rats using cell type-specific fiber photometry and optogenetic manipulation. Using chemogenetics, we found that motivation for a reward increased when orexin neurons were activated. Furthermore, during motivated behavior, orexin neuron activity changed dynamically: Activity increased during reward prediction and decreased after reward receipt. When an unexpected event occurred (i.e., not obtaining an expected reward), increased orexin activity was sustained. Notably, orexin activity strengthened with increasing effort. Optogenetic inhibition of orexin neuron activation during reward prediction and treatment with an orexin 1-receptor antagonist reduced reward-seeking behavior. Therefore, orexin is crucial for linking predicted expectations with motivated behavior. Moreover, optimization of orexin activity is necessary to overcome difficulties during motivated behaviors.
: Growth hormone deficiency (GHD) can lead to significant growth and developmental issues, necessitating long-term treatment. Once-weekly somatrogon is a long-acting recombinant human growth hormone analogue, approved for treatment of children with GHD. : Assess the long-term safety and efficacy of somatrogon in children with GHD. : Open-label extension (OLE) study following an initial 12-month open-label phase 2 study. : Conducted at 14 centers across Hungary, Bulgaria, Belarus, Ukraine, Russia, Greece, and the US. : Fifty-three children with GHD (67.9% male, mean age 6.0±2.1 years) completed the phase 2 study and 48 entered the OLE. : Participants received once-weekly somatrogon for up to 9 years. : Safety endpoints included adverse events (AEs), antidrug antibodies (ADAs), local site injection reactions, and insulin-like growth factor-I SDS. Efficacy endpoints included annual height velocity (HV), change in height SDS, and annual bone maturation. Treatment-emergent AEs (TEAEs) incidence was 52.1% at OLE year (Y) 1, decreasing to 23.8% by Y9. Serious AEs were rare with only isolated cases reported. ADA positivity was observed, but no neutralizing antibodies were found. No correlation between ADA presence and TEAE incidence or severity was noted, nor was there a difference in growth based on ADA presence. Mean annualized HV remained > 5 cm/year, and height SDS improved from -4.0 at baseline to -0.2 by Y8. No changes in the ratio of bone age/chronological age were observed. Once-weekly somatrogon is a well-tolerated and a viable long-term treatment for GHD in children. More real-world data are needed. Clinicaltrials.gov: NCT01592500.
Preexposure prophylaxis (PrEP) is a key biomedical HIV prevention strategy that relies heavily on adherence for optimal effectiveness. In China, most PrEP users purchase their medication online, making it challenging to monitor and support adherence effectively. This protocol aims to describe a digital intervention developed to monitor and improve the medication adherence of real-world e-consumers of PrEP and an evaluation plan assessing its acceptability, feasibility, and effectiveness. The Real-Time Monitoring and Precision Intervention for HIV PrEP Adherence (REMOTE) trial is a parallel-group, open-label, online-delivered, active-controlled, and stratified randomized controlled trial conducted among 430 e-consumers of PrEP (320 event-driven regimen users and 110 daily regimen users) in China. People who have purchased PrEP online, have taken PrEP in the previous 3 months, and plan to continue for the next 6 months will be stratified by regimen type and randomized into control and intervention groups at a 1:1 ratio. A WeChat-based digital platform will deliver the REMOTE intervention. The monitoring module follows ecological momentary assessment principles. The intervention design is guided by the stages of change theory, tailoring strategies to different nonadherence risks based on monitoring results. Intervention components include low-risk (health education and an artificial intelligence chatbot), medium-risk (peer forum and admonitory education), and high-risk (customized reminders and physician counseling) strategies. The control group will use a simplified platform with only real-time monitoring. The primary outcome is the proportion of participants achieving optimal adherence for 6 months, assessed via real-time monitoring and validated via surveys at baseline and the 1-, 3-, and 6-month follow-ups. Secondary outcomes include PrEP adherence knowledge, self-efficacy, risk perception, adherence barriers, stigma, and social support, measured via surveys. Intention-to-treat analysis will be conducted. Funding for the study was approved in March 2024. Ethics approval for the study was granted in July 2024. The pilot trial was completed in November 2025. Baseline data collection commenced in January 2026. By February 5, 2026, recruitment and baseline data collection were completed, with 448 participants enrolled. The data have not been viewed by the research team. The intervention is currently ongoing, and the study is expected to conclude in August 2026. Results are anticipated to be published in early 2027. The REMOTE trial pioneers a real-time monitoring and precision intervention for e-consumers of PrEP. Leveraging technology and ecological momentary assessment, it delivers a personalized, real-time intervention that is crucial for adherence. The findings could significantly impact future HIV prevention strategies. Chinese Clinical Trial Registry ChiCTR2400088278; https://www.chictr.org.cn/showproj.html?proj=236414. DERR1-10.2196/92750.
The ability to dynamically adjust a behavioral response to a stimulus depending on context is of critical importance for animals. To investigate the neural basis supporting context-dependent sensory processing, we developed a behavioral task in which mice changed their response to a single whisker deflection according to a continuously present contextual cue. Through unbiased optogenetic inactivation mapping, we found that neuronal activity in sensory and motor cortices contributed to task execution and, interestingly, we uncovered an unexpected role of the retrosplenial cortex (RSC) for contextual integration. Widefield calcium imaging revealed that the RSC was the first dorsal cortical area to show context discrimination in response to whisker stimulation, followed by the whisker motor cortex. Finally, we combined optogenetic inactivation with calcium imaging to define causal context-dependent changes in sensorimotor processing. Our cortex-wide mapping experiments thus begin to define key cortical nodes for context-dependent sensorimotor transformation and highlight an important contribution of RSC.
This study investigated the color-related compounds of Salvia miltiorrhiza after sweating. Stereomicroscopy and Electronic eye system were employed to characterize color variation. UPLC-Q-Orbitrap-MS and HPLC identified differential compounds. Furthermore, μ-FTIR and MALDI-MSI visualized the spatial distribution of compounds, and the compounds distribution was confirmed via HPLC. Result showed that after sweating, the appearance characteristics of Salvia miltiorrhiza changed significantly, exhibiting a reddish-brown epidermis and purple-brown inner tissue. Among 49 significantly altered compounds, tanshinones accounted for 58%. HPLC further indicated that the content of tanshinones showed a significant increase. Integrated μ-FTIR and MALDI-MSI demonstrated tanshinones increased after sweating and accumulated in the epidermis. Further confirmation of the increase in tanshinones was achieved by HPLC analysis of epidermis and cortex isolated. These results reveal tanshinones as the color-related compounds and clarify their spatial distribution, providing insights into the chemical changes during the traditional sweating process of Salvia miltiorrhiza.
Safety studies of the COVID-19 vaccine have identified some adverse events. Yet newer variant-updated formulations, along with increased hybrid immunity, may change these risks. Early-era safety data may not reflect experience with updated formulations in more immune-experienced populations. To evaluate 90-day risks for adverse events after coadministration of COVID-19 and influenza vaccines compared with influenza vaccination alone, across bivalent, XBB-adapted, and KP-adapted COVID-19 vaccine periods. Target trial emulation using electronic health care data. U.S. Department of Veterans Affairs. Participants receiving both COVID-19 and seasonal influenza vaccines (n = 705 124) and those receiving only an influenza vaccine (n = 1 813 205) between 1 September 2022 and 26 August 2025. Receipt of both COVID-19 and seasonal influenza vaccines versus receipt of only an influenza vaccine. 90-day risks for 46 prespecified individual adverse events grouped into 3 composite outcomes (tier 1, serious or life-threatening; tier 2, clinically significant; tier 3, less severe or self-limiting), using weighted discrete-time survival models. For all 3 composite outcomes, risks were similar between groups: tier 1 (risk ratio [RR], 1.03 [95% CI, 0.99 to 1.09]), tier 2 (RR, 0.99 [CI, 0.96 to 1.03]), and tier 3 (RR, 0.99 [CI, 0.96 to 1.02]). Of the 46 individual adverse events, 2 tier-3 risks had nominal statistical significance: syncope (RR, 1.09 [CI, 1.02 to 1.17]) and tinnitus (RR, 0.95 [CI, 0.92 to 0.99]); no risks were statistically significant after correcting for multiple comparisons. For all risks in tier 1 or tier 2, confidence bounds included 1.0 (no effect). In period-stratified analyses, neither composite (tier) nor individual event estimates supported differences in risks between groups. Generalizability and potential unmeasured confounding. Same-day coadministration of COVID-19 and influenza vaccines was not associated with an increased risk for adverse events in 3 updated-formulation periods. These findings support the short-term safety of coadministration. U.S. Department of Veterans Affairs.
Bone is a major mechanosensitive organ that continuously responds to physical cues in the body. Although mechanical stimulation plays important roles in skeletal development, homeostasis, and injury repair, the specific responses of skeletal stem cells (SSCs) to mechanical stress remain incompletely understood. To address this question, we established an in vitro model to isolate mouse periosteal SSCs and apply mechanical stimulation. Mouse periosteal cells were first obtained by enzymatic digestion, and SSCs were subsequently isolated by fluorescence-activated cell sorting (FACS) based on established surface markers. After attachment to culture, SSCs were subjected to tensile mechanical stimulation using a cyclic cell stress-tension system. This system enables the reproducible application of tensile force to cultured cells and provides a platform for analyzing SSC responses to mechanical input. Using this approach, we found that the expression of the cellular senescence-associated genes p16 and p21 was markedly reduced in SSCs following mechanical stimulation. These findings suggest that tensile stimulation may influence senescence-related changes in SSCs under in vitro conditions. Overall, this protocol provides a useful and suitable platform for studying SSC behavior under mechanical stimulation and for investigating how mechanical cues affect SSC function.
Early-life exposure to ionizing radiation is a recognized risk factor for pediatric brain tumors, including medulloblastoma (MB), but the dose-dependent mechanisms linking radiation to tumor initiation remain poorly understood. Using the Ptch1⁺/⁻ mouse model of Sonic Hedgehog (SHH)-driven MB, mice were exposed at postnatal day 2 to 0.1 or 2 Gy γ-rays and monitored longitudinally, combined with multi-omics analyses to characterize early cerebellar responses and tumor-associated transcriptional programs. Radiation increased MB incidence in a dose-dependent manner, with 2 Gy significantly increasing tumor burden and reducing latency, whereas 0.1 Gy elevated tumor risk without affecting onset. Genome-wide methylation profiling revealed progressive dose-dependent epigenetic remodeling, with broader developmental and transcriptional regulatory changes after high-dose irradiation and limited signaling-associated alterations after low-dose exposure. Proteomic analyses identified common proliferative programs together with distinct dose-specific biological contexts, with low-dose irradiation preferentially associated with regulatory, transport, and selected immune-related pathways and high-dose exposure with chromatin organization, DNA replication, nucleotide metabolism, and RNA-associated processes. Radiation-induced MBs exhibited transcriptional architectures distinct from spontaneous tumors and differed according to radiation dose. Although involving largely distinct genes and pathways, independent methylomic, proteomic, and transcriptomic analyses converged on the identification of qualitatively distinct dose-associated biological programs. Notably, cross-omics functional concordance increased with radiation dose, revealing greater integration of molecular responses following high-dose irradiation. Together, these findings define dose-specific molecular states associated with radiation-induced MB and provide a biological framework linking early cerebellar remodeling to tumor heterogeneity, with implications for pediatric radiation risk assessment and therapeutic stratification.