The Journal of Headache and Pain

Background: Post-dural puncture headache (PDPH) affects up to 11.2% of patients after neuraxial anesthesia. The sphenopalatine ganglion block (SPGB) is a promising minimally invasive intervention, but high-quality randomized trial data are limited. We conducted a pilot randomized controlled trial to assess feasibility and inform a future definitive trial. Methods: Twenty-six patients with PDPH following accidental dural puncture with 17G Tuohy needles were randomized to conservative management (bed rest, hydration) or SPGB (bilateral intranasal 2% lidocaine). Primary outcomes were feasibility (recruitment, retention, protocol adherence). Secondary outcomes included pain intensity (Numeric Rating Scale, NRS 0-10) at 30 minutes, 12 hours, and 24 hours; rescue analgesia requirements; mobilization time; and adverse events. Results: Feasibility was confirmed: 100% recruitment of target sample, 100% retention, 100% protocol adherence. At 30 minutes, all SPGB patients reported complete pain resolution (NRS=0) versus median NRS 3 (IQR 2) in controls (p<0.001), though this finding is limited by lack of blinding and baseline assessment. No SPGB patients required rescue analgesia or experienced adverse events. Conservative group patients had prolonged hospitalization (46%). Sample size calculation for a definitive trial (90% power, =0.05) yields 120 participants (60/group). Conclusions: A definitive RCT comparing SPGB to conservative management for PDPH is feasible. Preliminary efficacy data suggest rapid analgesia with SPGB, but rigorous confirmation in a sham-controlled trial is required. Trial registration: ClinicalTrials.gov -NCT07494383 (retrospectively registered). Keywords: Post-dural puncture headache, sphenopalatine ganglion block, pilot study, feasibility, regional anesthesia, randomized controlled trial

Background. Calcitonin gene-related peptide (CGRP)-targeted therapies, including injectable monoclonal antibodies (mAbs: erenumab, fremanezumab, galcanezumab, eptinezumab) and oral gepants (atogepant, rimegepant), represent a paradigm shift in episodic migraine prevention. No direct head-to-head trials across the full drug class exist. We conducted a PRISMA-NMA-compliant Bayesian network meta-analysis (NMA) to compare the relative efficacy and tolerability of all approved CGRP-targeted preventive therapies. Methods. PubMed, Embase, and Cochrane CENTRAL (inception to January 2026) were searched for doubleblind RCTs in episodic migraine. A Bayesian random-effects NMA used Markov Chain Monte Carlo simulation. Primary outcome: change in monthly migraine days (MMD). Secondary outcomes: 50% or greater responder rate, TEAEs, and DAEs. SUCRA probabilities quantified treatment rankings. Transitivity was formally assessed. Publication bias was evaluated using comparison-adjusted funnel plots and Egger test. GRADE certainty was rated for all key comparisons. Results. Thirty-two RCTs (24,418 participants; mean age 39.2 years; 84% female; mean baseline 8.2 MMD) were included (Table 1). All active treatments significantly reduced MMD versus placebo. Eptinezumab 300 mg ranked highest for MMD reduction (MD 2.40 MMD, 95% CrI 3.10 to 1.70; SUCRA 91.2%), followed by galcanezumab 240 mg (SUCRA 85.4%) and erenumab 140 mg (SUCRA 79.8%). For the 50% responder rate, galcanezumab 240 mg ranked highest (OR 3.12, 95% CrI 2.22 to 4.38; SUCRA 92.1%). Oral gepants demonstrated significant but more modest efficacy: atogepant 60 mg (SUCRA 38.4%) and rimegepant (SUCRA 28.9%). The absolute mAb-versus-gepant efficacy difference of approximately 1.1 MMD exceeded the accepted minimal clinically important difference. Gepants demonstrated placebo-comparable tolerability (TEAE RR 1.02, 95% CrI 0.93 to 1.12; SUCRA 93 to 96%). Heterogeneity was low to moderate (I-squared 14 to 31%); no significant network inconsistency (node-split p greater than 0.29); and no significant publication bias (Egger test p = 0.24). GRADE certainty was high for class-versus-placebo comparisons and moderate for indirect mAb-versus-gepant comparisons. Conclusion. CGRP mAbs provide superior efficacy over oral gepants for episodic migraine prevention. Oral gepants offer placebo-comparable tolerability. An individualized, patient-centered approach guided by symptom burden, comorbidities, administration preference, and the efficacy-tolerability tradeoff of each drug class is recommended.

Inflammatory bowel disease (IBD) is characterised by chronic pain, a debilitating symptom for which effective treatments are few and far between. IBD pathogenesis includes the prevalence of a variety of pro-inflammatory cytokines, including the Interleukin-6 (IL-6) family members Il-6 and Oncostatin M (OSM). Previous research has shown disruption of OSM signaling can modulate nociceptor sensitization and activation, however the downstream signalling pathway is unknown. When an in silico analysis of murine colonic sensory neuronal populations was undertaken for receptor expression for OSM and other factors necessary for intracellular signaling, we can find diverse expression indicative of functional signaling. We were able to observe that hyper Il-6 (Il-6 bound to the soluble Il-6 receptor) and OSM can elicit activation of a subset of murine sensory neurons by finding an increase in calcium mobilization following superfusion. This could then be attenuated by the pharmacologic inhibition of all janus kinases or interestingly, TYK2 alone. Furthermore, inhibition of transient receptor potential vanilloid 1 or transient receptor potential ankyrin 1 ion channels, which are known to be sensitized by OSM in other sensory neurons also reduced the proportion of OSM-responsive neurons. This further understanding of OSM signaling in sensory neurons creates avenues for more extensive research into the molecular mechanisms occurring as well as the potential to exploit these therapeutically to induce analgesia in a subset of neurons.

Background Migraine prevalence is higher among individuals with attention deficit hyperactivity disorder (ADHD). However, most research has focused on single-disease studies. This study used Global Burden of Disease (GBD) data to analyze co-occurrence patterns and related risk factors. Methods This study extracted the incidence and age-standardized incidence rate (ASIR) of migraine and ADHD among individuals across 204 countries and territories in GBD 2021, as well as exposure values for risk factors. To explore the co-occurrence patterns of migraine and ADHD and their spatial heterogeneity in global distribution, the incidence of both diseases was classified into quartiles, and countries and territories were categorized into three regional types: consistent regions, migraine-dominant regions, and ADHD-dominant regions. Global groupings by economy and risk factors were analyzed separately for co-occurrence patterns, and disease burden projections were made for 2050. Results In 2021, countries and regions were categorized into three distinct groups based on disease prevalence patterns: the majority exhibited an ADHD-dominant profile, predominantly found in high-SDI regions; a consistent pattern, where both diseases occurred at comparable levels, was primarily observed across South Africa and the Middle East, while a migraine-dominant pattern was identified in North Africa. Co-occurrence patterns were generally less prevalent in areas with lower socioeconomic development. Across all three patterns, high temperature exposure, iron deficiency, and metabolic risks emerged as the primary contributing factors. Looking ahead to 2050, the global burden of migraine was projected to stabilize, whereas the prevalence of ADHD was expected to experience a slight yet consistent increase. Conclusion This study systematically identifies the co-occurrence patterns of ADHD and migraine, along with their socioeconomic and environmental drivers, offering evidence-based insights for early prevention and targeted intervention in disease populations globally. Keywords GBD, migraine, ADHD, incidence, disease burden

Previous brain decoding studies indicate that an individuals pain experience can be robustly predicted from distributed patterns of brain activity. Two brain decoders have notably been associated respectively with the nociceptive and cognitive aspects of pain experience, the Neurologic Pain Signature (NPS) and the Stimulus-Intensity Independent Pain Signature (SIIPS). Yet, we still do not know if these brain patterns are also causally related to pain experience. To evaluate this possibility, we used high-field (7-Tesla) fMRI to test whether humans can alter their pain experience by bidirectionally modulating their pain-related brain activity in decoded neurofeedback paradigm. In a double-blind design, participants were trained to up- and down-regulate the NPS or the SIIPS. Our results indicate that participants can achieve bidirectional control of both signatures. NPS expression reliably increased during pain stimulation and covaried with both stimulus intensity and subjective ratings. In contrast, SIIPS expression did not show consistent stimulus-locked effects in the primary analyses. Importantly, reduction in pain rating was specific for SIIPS-training, whereas NPS has failed to show any consistent behavioral effect. Based on these preliminary findings, we hereby preregister a follow-up study, with specified rationale, hypotheses, experimental design, and analysis protocols.

The severity stratification of carpal tunnel syndrome (CTS) relies on ultrasound morphological markers and electromyography. However, it remains unclear how structural imaging can reliably infer functional impairment. Clarifying the structure-function relationship is critical for efficient diagnostic pathways. A retrospective cohort of 55 patients with symptoms related to CTS was analyzed at the Shanghai Sixth Peoples Hospital. All patients were subjected to ultrasound and EMG. 72.7% cases were diagnosed with CTS with a female predominance and equal left-right involvement. Random-forest classifiers were trained using surrogate splits, and performance was evaluated using predictions outside the bag. A full-feature model (34 candidate variables) was compared against a simplified model (8 core variables) capturing the core morphological and electrophysiological features. A residual-based framework was then used to characterize the structure-function mismatch within severity grades (1a-3c). The simplified model improved discriminative performance compared to the full-feature model (AUC 0.789 to 0.824). The simplified model achieved an overall accuracy of 77.3%. Analysis of predicted probability distributions and 10-bin calibration curves indicated stable and clinically interpretable risk estimation in most probability ranges. Permutation-based importance analysis confirmed that both ultrasound and electrophysiological features contributed substantively to prediction. Residual-based grading further revealed structure- function heterogeneity within each main severity grade. CTS severity can be stratified using a limited set of complementary morphological and electrophysiological features. Structure-function mismatch supports an imaging-led initial screening, with electrophysiology reserved for selected patients.

Nerve injury causes an imbalance of glutamatergic excitation over GABAergic inhibition, contributing thereby to lasting neuropathic pain. Transgenic GAD67-GFP knock-in reporter mice were developed to visualize GABAergic interneurons. The knock-in into glutamate decarboxylase (GAD67) causes haploinsufficiency that manifest in low GABA levels. In this model, we studied if diminished GABA exacerbates neuropathic pain after nerve injury. Adolescent male and female GAD67-GFP knock-in mice were subjected to Spared Sciatic Nerve Injury (SNI). At baseline, nociception and thermal preferences were equal but after SNI, GAD67-GFP mice developed thermal allodynia which was not detected in wildtype littermates. At the electrophysiology level, SNI caused a partial decrease in the excitability in layer 2/3 pyramidal neurons in the projection-hemisphere in wildtype mice. This effect was exacerbated in GAD67-GFP, affecting both sides, and was accompanied with imbalance of field-potential (FP) amplitudes between projection and non-projection hemisphere, which did not occur in wildtype mice. The results suggest that GABA deficiency can be compensated to protect from hyperexcitability at baseline, but it cannot be further upscaled, ultimately leading to network hyperactivity after injury. Metabolomic studies confirmed the moderate loss of GABA in ipsi- and contralateral cortex and lumbar spinal cord of GAD67-GFP mice and failure to raise GABA in the ipsilateral dorsal horn after injury. Carnosine, cystathionine, and glutathione, three important anti-oxidative metabolites, were co-reduced with GABA suggesting that GABAergic activity and anti-oxidative capacity are interconnected and failure of this axis contributes to neuropathic "pain".

Deficient descending pain inhibition assessed by conditioned pain modulation (CPM) is considered a common feature of various chronic pain disorders. Typically, CPM studies focus on one particular disorder making direct comparisons between disorders difficult. This cross-sectional study aimed to compare CPM effects between three clearly distinct chronic pain disorders and pain-free controls. Furthermore, patients were pooled with controls to explore whether subgroups showing different CPM effects could be separated independent of cohort membership. One hundred and forty participants (patients: 53 non-specific chronic low back pain [nsCLBP], 15 complex regional pain syndrome [CRPS], 14 neuropathic pain after spinal cord injury [painSCI]; 58 controls) were included. CPM effects were assessed in a remote, pain-free area using pressure pain thresholds as test stimulus and a cold water bath as conditioning stimulus. Cohort differences in CPM effects were analyzed using linear mixed models. The presence of subgroups showing different CPM effects was tested using latent class linear mixed models. CPM effects differed between cohorts (p = 0.011), driven mainly by reduced inhibitory CPM effects in patients with nsCLBP compared to patients with painSCI. Latent class analysis detected 3 subgroups with varying degrees of significant inhibitory CPM effects (ps [&le;] 0.002). All subgroups comprised patients and controls. These results oppose deficient descending pain inhibition as a common feature of chronic pain disorders. Additionally, the failure to identify subgroups without inhibitory CPM effects within a heterogenous patient/control sample challenges the utility of deficient CPM as predictor of chronic pain or treatment efficacy. PerspectiveInhibitory conditioned pain modulation, a measure of descending pain inhibition, is not consistently impaired across distinct chronic pain disorders. Furthermore, identifying individuals with impaired conditioned pain modulation within a heterogenous sample is difficult. Thus, for conditioned pain modulation to be clinically useful, its variability needs to be better understood.

Neuropilin-1 (NRP1) is a single pass transmembrane glycoprotein that can form a receptor complex with several tyrosine kinase receptors, including the vascular endothelial growth factor (VEGF) receptor. Previous studies have reported that binding of VEGFA to this receptor complex elicits mechanical allodynia and thermal hyperalgesia through potentiation of voltage-gated sodium and calcium channel activity. We find that Nrp1 mRNA and protein is widely distributed in naIve mouse and rat DRG neurons, including peptidergic afferents. A CGRPcreER: NRP1fl/fl transgenic mice was generated to investigate the role of peptidergic NRP1 in basal nociception. Following in vivo loss of NRP1, mice are hyposensitive to both noxious heat and mechanical stimuli. Under normal conditions, VEGFA elicits mechanical hypersensitivity, an effect that was absent in our NRP1 knockout mouse. Furthermore, VEGFA induced neuronal hyperexcitability was lost in CGRP expressing neurons isolated from this NRP1 knockout mouse. This study validates the NRP1 knockout mouse and confirms previous findings that VEGFA, often released during pathological pain conditions, requires peptidergic NRP1. Interestingly, we find that in the absence of ongoing injury or inflammation, peptidergic NRP1 regulates basal nociception and pain-like behaviors. PerspectiveNRP1 is expressed in sensory neurons including the peptidergic subpopulation. Genetic deletion of NRP1 in healthy adults alters nociception without altering innervation; NRP1 knockout mice are hyposensitive to noxious heat and mechanical stimuli, but lose sensitivity to VEGFA, confirming it is a therapeutic target for growth factor mediated pain conditions.

Background: Spinal pain, including neck pain and low back pain (LBP), is a common musculoskeletal condition and major contributor to disability worldwide. Evidence comparing disability, fatigue and mental health across acute and chronic stages remains limited, particularly in conflict-affected and low-resource settings. This study assessed these outcomes among patients with acute and chronic neck pain and LBP in the Gaza Strip. Methods: A comparative cross-sectional study was conducted among 410 adults attending outpatient physical therapy at Nasser Medical Complex, Khan Younis, Gaza Strip. Participants included 204 with neck pain and 206 with LBP, classified as acute neck pain (n=101), chronic neck pain (n=103), acute LBP (n=102) and chronic LBP (n=104). Disability, fatigue, psychological distress and sleep disturbance were assessed using the Neck Disability Index (NDI)/Oswestry Disability Index (ODI), Fatigue Severity Scale (FSS), Patient Health Questionnaire-4 (PHQ-4) and PROMIS Sleep Disturbance Short Form 8a. Independent t-tests, adjusted linear regression, correlation analyses, clinical-threshold analyses and binary logistic regression were performed. Results: Chronic neck pain and chronic LBP were associated with significantly higher disability, fatigue and psychological distress than acute pain. Chronic neck pain patients had higher NDI, FSS and PHQ-4 scores than acute neck pain patients; chronic LBP patients had higher ODI, FSS and PHQ-4 scores than acute LBP patients (all p<0.001). Sleep disturbance did not differ significantly between groups. Female participants reported higher psychological distress in both pain groups, with higher fatigue in neck pain and higher disability in LBP. Adjusted analyses confirmed that chronic pain status remained associated with higher disability, fatigue and psychological distress. Fatigue was the most consistent factor independently associated with chronic pain status. Conclusions: Chronic spinal pain was associated with greater disability, fatigue and psychological distress than acute spinal pain, while sleep disturbance was common across groups. These findings support early multidimensional assessment, including screening for fatigue and psychological distress. Longitudinal studies are needed to clarify whether these factors contribute to transition from acute to chronic spinal pain.

BackgroundNeuropathic pain is a major source of disability and distress with few pharmacological options for treatment. Opioid drugs can be effective, but high doses are needed, leading to unwanted effects. BMS-986122 is a positive allosteric modulator of the mu opioid receptor that potentiates acute opioid antinociception without increasing opioid-induced constipation, reward, or respiratory depression. Therefore, we asked if BMS-986122 could increase the effects of low-dose opioid analgesics in chronic neuropathic pain. MethodsWe employed the spared nerve injury and tibial neuroma models in rats and assessed the tactile hypersensitivity of the hind paw and site of neuroma, respectively. ResultsAdministration of low doses of (R)-methadone, morphine, or buprenorphine slightly reduced the tactile hypersensitivity of the hind paw the in spared nerve injury model. Pretreatment with BMS-986122 significantly enhanced the reversal of hypersensitivity, reaching the effect of high-dose gabapentin, a standard of care in neuropathic pain. Pretreatment with BMS-986122 similarly increased the anti-allodynic effects of low dose (R)-methadone on neuroma pain. A similar effect of (R)-methadone in the absence of BMS-986122 was only observed at a dose where respiratory distress was seen. ConclusionsThese findings show that allosteric modulators of the mu opioid receptor such as BMS-986122 can enhance opioid activity that could translate to a safe and effective treatment for chronic neuropathic pain.

Traumatic brain injury (TBI) is a condition of high incidence worldwide, but remains mostly undertreated. Previous observations in preclinical studies pointed to a beneficial effect of insulin-like growth factor 1 (IGF-1) in TBI. As brain injury is associated to loss of IGF-1 sensitivity, we tested the therapeutic potential of AIK3a305 (AIK3), a novel IGF-1 sensitizer. Twenty-four hours after mild TBI induced by controlled impact, mice received daily intraperitoneal injections of AIK3 during 4 weeks. We found that TBI-associated sensorimotor disturbances measured with the adhesive-removal test were reverted by AIK3 treatment. In addition, neurological and cognitive disturbances measured by the neurological severity score and Y maze respectively, were also ameliorated by treatment with the IGF-1 sensitizer, whereas increased anxiety after mild TBI was also normalized by AIK3. Circulating levels of IGF-1 were increased after AIK3 treatment in TBI mice, while serum IL-6 levels, a biomarker of inflammation associated to TBI were similar to control mice treated with AIK3. Transcriptomic analysis determined that treatment with AIK3 widely affected gene expression in TBI brains, showing a general reduction in both up- and down-regulated genes. Collectively, these data support the use of IGF-1 sensitizers such as AIK3 for treatment of TBI.

IntroductionVagus nerve stimulation modulates laryngeal, cardiac, pulmonary, and gastrointestinal functions. Knowledge of where along the vagal trunk organ-specific branches emerge may support alternative surgical placements of stimulation devices to engage targeted functions while avoiding off-target effects. However, no quantified map of how vagal branches emerge and how they relate to surgically relevant anatomical landmarks exists in humans. MethodsFifty-eight vagus nerves (29 left, 29 right) from 29 embalmed donor bodies (15 females) were dissected from the jugular foramen through the thoracic cavity. Branches were traced to end organs and allocated to seven groups -- sympathetic, muscular, vascular, cardiac, pulmonary, esophageal, and multiple targets -- and several sub-groups. Distances between branch emergence and the jugular foramen (JF) were normalized to three anatomical landmarks: carotid bifurcation, laryngeal prominence, and superior border of clavicle. ResultsBranch emergence follows a proximal-to-distal order: sympathetic (5.28 cm from JF), muscular (9.59 cm), vascular (10.70 cm), cardiac (19.65 cm), pulmonary (25.36 cm), and esophageal (26.57 cm). Vagal branches emerge into two embryological domains separated near the clavicle: pharyngeal arch-targeting branches cluster proximally (9.34 cm) and primitive mediastinum-targeting branches cluster distally (23.74 cm), with sympathetic, muscular, and vascular sub-groups occupying distinct zones within the proximal domain. The largest branch-free intervals occur above the left clavicle (2.33 {+/-} 2.80 cm) and below the left carotid bifurcation (2.58 {+/-} 3.17 cm). Alternate placement regions separating targeted organs from off-targets: sympathetic vs. cervical visceral at 6/8 cm (L/R), cardiac vs. carotid sinus/bifurcation at 14/10 cm, and recurrent laryngeal vs. other cervical visceral at 18/13 cm from JF. Overall, no differences were found between male and female donors. ConclusionsThis study provides a quantified, landmark-registered map of cervical and thoracic vagal branch emergence, offering a standardized anatomical template that may inform strategies for more function-selective vagal neuromodulation.

PurposeBacterial vaginosis (BV) is associated with disruption of the vaginal microbiome and extracellular matrix (ECM) remodeling, yet the contribution of host proteases to this process remains unclear. This study investigated whether expression and activity of cathepsins K, L, S, and V differ by BV diagnosis and community state type (CST). We hypothesized that BV and BV associated CSTs would exhibit increased expression and activity of collagen and elastin-degrading cathepsins. MethodsVaginal fluid samples were collected and classified by BV diagnosis and CST. Cathepsin expression was evaluated by Western blotting to distinguish inactive and active enzyme forms. Proteolytic activity was assessed using multiplex cathepsin zymography. Statistical analyses compared cathepsin expression and activity across diagnoses and CSTs. Principal component analysis and linear regression were performed to assess associations between cathepsin activity, microbial diversity, and CST. ResultsProcathepsin K expression was significantly increased in BV-positive and CST IV samples, while total cathepsin L expression was significantly elevated in samples with Nugent-intermediate scores. Cathepsins S and V showed variation in inactive and active forms in Nugent-intermediate and CST III samples. In contrast, total cathepsin activity, including cathepsins K and V, did not significantly differ across BV diagnoses or CSTs. Overall, cathepsin activity varied between individuals rather than by clinical classification. ConclusionsCathepsin expression and maturation state differ by microbiome composition, suggesting that the vaginal microbiome may regulate post-translational processing of cathepsins. As a result, cathepsin activity appears to be regulated at the individual level rather than strictly by BV diagnosis or CST. These findings link vaginal microbiome composition to ECM remodeling and potential adverse reproductive outcomes.

More than 100,000 people in the UK have an ischaemic stroke every year. This means a blood clot blocks an artery supplying blood to their brain. Patients with ischaemic stroke often have sudden weakness affecting half of their body or face, and problems with speech, although other stroke symptoms are possible. Treatment with a clot-busting drug (thrombolysis) can reduce the amount of disability and death caused by ischaemic stroke by restoring the blood supply to the brain. Thrombolysis usually needs to be given within four and a half hours of stroke occurring. Unfortunately, nearly three quarters of patients with ischaemic stroke arrive in hospital later than this, or it is not clear when their stroke started. For these patients, it may still be possible to treat them with thrombolysis if the hospital can provide an additional advanced type of brain scan. However, many hospitals in the UK and worldwide cannot offer this advanced scan to patients with stroke, particularly hospitals that are not in major cities or developed nations. Patients arriving at these hospitals therefore do not currently have the same access to effective treatment for stroke. We have developed a simple method for identifying which patients can be given thrombolysis even if they arrive at hospital later than four and a half hours, or where there is uncertainty about when their stroke started. Our method does not require any additional or advanced imaging, only the standard computed tomography (CT or CAT) scan that all patients with stroke get when they arrive at hospital. Our method is called the CT Clock. We ask doctors to look for stroke changes indicating ischaemic stroke in the brain on CT. If they find these stroke changes, they measure them compared to normal brain. If the stroke changes on CT are minor (less than 20% darker than normal brain), or the scan appears normal despite quite severe stroke symptoms, we would consider these patients suitable for treatment with thrombolysis. This study proposes to test the safety of our CT Clock method in an analysis of existing NHS data from patients who have previously been treated with thrombolysis. We aim to show that when doctors who provide stroke care use our CT Clock method to identify suitable patients for treatment with thrombolysis, it is safe for these patients. If successful, this study will allow us to plan for and deliver future clinical testing where we would use the CT Clock to identify patients for treatment with thrombolysis in hospitals where advanced imaging for stroke is unavailable. Successful clinical testing is needed before our method can be used routinely in hospitals around the world. Data Study Protocol O_TBL View this table: org.highwire.dtl.DTLVardef@18a7b6corg.highwire.dtl.DTLVardef@ad2388org.highwire.dtl.DTLVardef@f7ea61org.highwire.dtl.DTLVardef@3cc038org.highwire.dtl.DTLVardef@6faf1e_HPS_FORMAT_FIGEXP M_TBL C_TBL

BackgroundCurrent microphysiological models do not support long-term investigations into the chronic effects of vascular risk factors and the development of vascular diseases. Prolonged culture frequently leads to cellular senescence and loss of functional integrity, resulting in variability and inconsistency in modeling chronic vascular responses. Here we aimed to develop and sustain a long-term multicellular human vascular avatar, addressing the critical need for long-term disease modeling and drug testing. MethodsTo identify the optimal media for longevity, cell identity and function were assessed by morphology, qPCR, beta-gal staining, ELISA, bulk RNA-seq and single cell RNA-seq analysis. After optimizing the culture media, iPSCs-derived ECs and VSMCs from unaffected and Hutchinson-Gilford Progeria Syndrome (HGPS) donors were grown in Gravitational Lumen Patterning (GLP) Vessel- Chips for 1-6 months to generate a long-lived vascular avatar for the study of vascular aging. ResultsGuided by quantitative morphological analyses and bulk RNAseq profiling, we generated a novel optimized culture media VSL (VEGF, SB431542 as a TGF-{beta} inhibitor, low fetal bovine serum) that enhances the long-term health of vascular endothelial cells (ECs). Furthermore, we modified the VSL formulation (mVSL) by modulating 8Br-cAMP, FGF, PDGF, and a cell viability enhancer HMH1015 levels to enhance EC-VSMC (vascular smooth muscle cell) crosstalk and support long-term cellular viability. Subsequently, we maintained and characterized a human vascular avatar with a three-dimensional extracellular matrix environment and 3D vascular architecture for over 180 days. Finally, we demonstrated that this long-lived human vascular avatar enabled modeling vascular aging using iPSC-derived vascular cells from patients with Hutchinson-Gilford Progeria Syndrome (HGPS). ConclusionsWe have successfully engineered and maintained a human vascular avatar for over 180 days. The vascular avatar provides a robust platform for modeling disease-associated vascular aging and for evaluating therapeutic strategies targeting chronic vascular disorders.

Adult hippocampal neurogenesis plays a central role in learning, memory formation, and adaptive neural plasticity, making it an attractive target for noninvasive neuromodulation strategies. Low-intensity focused ultrasound (LIFU) has emerged as a promising modality for modulating brain function, yet its effects on adult neurogenesis and the role of stimulation frequency remain incompletely understood. In this study, we evaluated whether transcranial LIFU applied to the dentate gyrus influences neurogenic and cognitive outcomes in a frequency-dependent manner. Adult rats received twice-weekly ultrasound stimulation for four weeks at 0.5, 1, or 5 MHz. Neurogenesis was assessed through BrdU incorporation and neuronal differentiation by BrdU/NeuN co-labeling, while expression of neurogenesis-associated markers (BDNF, FGF-2, and Sox-2) was quantified using qRT-PCR. Behavioral effects were examined using the novel object recognition task. Among the tested conditions, 0.5 MHz stimulation produced the most pronounced neurogenic response, with increased cellular proliferation in the dentate gyrus, elevated expression of neurogenic markers, and improved recognition memory relative to sham-treated animals. Higher stimulation frequencies yielded comparatively weaker effects. These findings identify stimulation frequency as a critical determinant of LIFU-driven neuroplastic responses and support the potential of focused ultrasound as a noninvasive approach for promoting hippocampal regeneration and functional recovery.

Multipotent mesenchymal stem cells (MSCs) including bone marrow stromal cells (BMSCs) have shown analgesic efficacy in recent years. Studies suggested that the therapeutic effect of MSCs was mediated by their secreted small extracellular vesicles (sEVs) mainly exosomes. The present study evaluated the antihyperalgesic effect of BMSC-related sEVs in a mouse model of neuropathic pain involving chronic constriction injury of the infraorbital nerve (CCI-ION). Our separation protocol generated EV particles mostly sized in the range of exosomes (30-170 nm) and express exosome marker proteins CD9, CD81, and Tsg101, suggesting their endosome origin. We show that intravenous injection of BMSC-related sEVs attenuated pain hypersensitivity induced by CCI-ION as indicated by decreased mechanical hypersensitivity (von Frey test) and reduced aversion to noxious stimulation (conditioned place avoidance test). The antihyperalgesic effect of sEVs was observed in both female and male animals, and the effect was dose-dependent. sEVs from NAIVE serum-treated BMSC cultures produced short-lasting antihyperalgesia in male but not female mice, suggesting a subtle sex difference. The antihyperalgesia of sEVs from BMSC culture was blocked by the pretreatment of the culture with GM4869, the antagonist of exosome secretion, suggesting that the effect was not related to other co-isolated soluble mediators but mediated by MSC-derived exosomes. Interestingly, the prior injury condition in which sEVs were isolated favors the pain-relieving effect of sEVs. sEVs isolated from the serum of BMSC-treated animals receiving tendon ligation (TL) injury attenuated hyperalgesia for 24 h, while sEVs from the serum of BMSC-treated NAIVE animals only attenuated hyperalgesia at 3 h after injection. sEVs from the BMSC culture treated with the serum of TL rats were antihyperalgesic, but sEVs from the BMSC culture treated with the serum of naive animals were ineffective. Our results indicate that BMSC-related sEVs produced antihyperalgesia similar to that produced by BMSCs. The results suggest that the interactions between BMSCs and injury conditions are crucially important for producing efficacious sEVs/exosomes and support that the effect of sEVs could be optimized by priming BMSCs with injury-related conditions.

Age related cerebral atrophy is one of the most prevalent radiological findings in ageing populations, yet its clinical significance particularly its correlation with specific neurological presenting symptoms remains insufficiently characterised in South Asian contexts. This retrospective cross sectional study was conducted at THQ Hospital Wazirabad and Chattha Hospital, Gujranwala, Pakistan over a six month period, enrolling 66 adult patients ([&ge;]40 years) who underwent non contrast computed tomography (CT) of the brain. CT scans were evaluated for Evans index, ventricular enlargement (graded 1 to 3), cerebral atrophy severity (graded 1 to 3), early ischaemic changes, and the hyperdense vessel sign. Presenting neurological symptoms headache, seizures, slurred speech, ataxia, and numbness were extracted from medical records and correlated with imaging findings using chi square tests, Spearmans rank correlation, and binary logistic regression in SPSS v31.0. The mean patient age was 52.1 to 14.3 years (range 35 83) with a male predominance (72.7%). Moderate to severe atrophy was present in 50.0% of patients. Seizures (74.2%), slurred speech (63.6%), and ataxia (62.1%) were the most prevalent symptoms. Significant positive correlations were found between atrophy grade and age (r = 0.72, p < 0.001), slurred speech (r = 0.48, p < 0.001), ataxia (r = 0.44, p < 0.001), and numbness (r = 0.39, p = 0.001). Headache showed no significant correlation with atrophy severity (p = 0.42). Logistic regression revealed that each one grade increase in atrophy severity raised the odds of motor/speech symptoms by 2.8 fold (95% CI: 1.6 to 4.9, p <0.001), independent of age. These findings support the integration of standardised CT based atrophy reporting into routine radiology practice for older adults, especially in resource limited settings where MRI is not readily accessible.

Alzheimers disease (AD) is a neurodegenerative disorder affecting more than 55 million people worldwide, with a diagnosis that remains predominantly clinical and frequently delayed. The electroretinogram (ERG) offers a non-invasive electrophysiological method for detecting retinal dysfunction associated with neurodegeneration; however, it remains unclear whether robust and reliable candidate biomarkers can be extracted from ERG signals beyond conventional amplitude- and latency-based parameters. Here we present a pilot study of a multi-domain signal processing framework applied to ERGs recorded from 46 participants (20 AD patients, 26 controls) with a handheld device (RETeval, LKC Technologies) using sinusoidal (1-50 Hz) and photopic ISCEV protocols. Five complementary techniques were implemented: (i) multiscale fuzzy entropy (MSFuzzyEn); (ii) FFT harmonic analysis; (iii) stimulus-response wavelet time-frequency coherence (WTC); (iv) a novel inter-cycle lag variant of sample entropy (SampEnT), introduced to isolate cycle-to-cycle retinal response consistency independently of stimulus periodicity; and (v) discrete wavelet transform (DWT) for energetic extraction of oscillatory potentials (OPs). Univariate comparisons (Mann-Whitney, Cliffs{delta} , Benjamini-Hochberg FDR) identified seven significant candidate biomarkers (q < 0.05), five with large effect size: AUCfast (|{delta}| = 0.546, q = 0.009), Slopevery-slow (|{delta}| = 0.554, q = 0.007), R14f (|{delta}| = 0.515, q = 0.031), SampEnT (|{delta}| = 0.504, q = 0.019) and WTCR,mean (|{delta}| = 0.531, q = 0.023); and two with medium effect size (OP_amp_sum, band_snr). A logistic regression classifier combining three candidate biomarkers, validated by leave-one-out cross-validation, achieved ROC-AUC = 0.858, sensitivity = 70.0% and specificity = 88.5% (n = 46). These proof-of-concept results demonstrate that multi-domain ERG analysis captures retinal temporal dysfunction signatures in AD that are inaccessible to standard clinical analysis, supporting further investigation of portable ERG devices as a source of non-invasive candidate biomarkers for early AD detection.