Pharmacology Research & Perspectives

Abstract Background: Acute postoperative pain affects more than 80% of surgical patients, with orthopedic lower limb procedures consistently associated with severe pain intensity and high opioid requirements. Preemptive analgesia with oral agents has been proposed to attenuate central and peripheral sensitization prior to surgical incision. Tapentadol, a dual-mechanism -opioid receptor agonist and norepinephrine reuptake inhibitor, and pregabalin, a voltage-gated calcium channel modulator, represent pharmacologically distinct premedication options; however, direct comparative data in this surgical context are lacking. This pilot randomized controlled trial aimed to compare the analgesic efficacy and safety of 72-hour oral premedication with tapentadol versus pregabalin in patients undergoing elective lower limb surgery under neuraxial anesthesia. Methods: In this double-blind, parallel-group pilot trial, 46 patients scheduled for elective lower limb surgery under neuraxial anesthesia were randomized equally to receive tapentadol 50mg orally every 12 hours (Group A, n = 23) or pregabalin 75mg orally every 24 hours (Group B, n = 23), initiated 72 hours before surgical incision. The primary outcome was postoperative pain intensity assessed using the Numeric Rating Scale (NRS, 0-10) at post-anesthesia care unit (PACU) arrival (T0) and at 30 (T1), 60 (T2), 90 (T3), and 120 (T4) minutes thereafter. Secondary outcomes included Verbal Rating Scale (VRS) scores, rescue morphine consumption, and safety. The primary longitudinal analysis used a linear mixed model (LMM) with Group, Time, and Group x Time interaction as fixed effects and a random intercept per patient; between-group contrasts at each timepoint were derived from estimated marginal means with Holm correction. Effect sizes are reported as Cohen's d. Results: All 46 patients completed the study with no missing data. Both groups were pain-free at T0 (NRS=0). Pain scores diverged progressively from T1 onward, with the pregabalin group reporting consistently higher NRS values at every time point. The LMM revealed a significant main effect of Time (F4,181.6 = 23.61, p < 0.001) and a borderline-significant Group x Time interaction in the continuous-time sensitivity model (F1,187.6 = 3.79, p = 0.053). Post-hoc contrasts identified a statistically significant, large effect between-group difference at T3 (mean NRS difference -0.91, p = 0.006, Cohen's d = -0.96) and a medium-effect trend at T2 (d = -0.59, p = 0.089). Rescue analgesia was required by 4.3% of tapentadol patients versus 21.7% of pregabalin patients. Nausea and vomiting were equally present in both groups (17.4%). No hypersensitivity reactions were observed in either arm. Conclusions: Seventy-two-hour oral premedication with tapentadol 100mg/day provided superior postoperative analgesia compared with pregabalin 75 mg/day at the 90-minute PACU time point, with a large effect size and a fivefold reduction in rescue analgesia requirements. Both agents were well tolerated. These pilot data support the conduct of a fully powered, multicenter randomized controlled trial to confirm the analgesic superiority of tapentadol premedication in orthopedic lower limb surgery.

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: Orthopedic surgeries are associated with significant intraoperative and postoperative pain, necessitating effective anesthesia strategies. Spinal anesthesia is commonly used for lower limb procedures due to its rapid onset and reliability; however, its limited duration may compromise prolonged surgical procedures and early postoperative pain control. Adjuvants such as dexamethasone have been explored to enhance and prolong the effects of local anesthetics. While evidence supports its efficacy, data from low-resource settings remain limited. Objective: To assess the effect of intrathecal dexamethasone as an adjuvant to bupivacaine on sensory block duration, time to first postoperative analgesia, and postoperative pain in patients undergoing lower limb orthopedic surgery at KCMC. Methodology: A randomized, double-blind controlled trial was conducted among 96 adult patients undergoing elective lower limb orthopedic surgery under spinal anesthesia. Participants were allocated using a computer-generated randomization sequence to receive either bupivacaine 15 mg with dexamethasone 4 mg (intervention group) or bupivacaine 15 mg with 1 ml normal saline (control group). Outcomes included sensory and motor block duration, time to first postoperative analgesia, and postoperative pain scores. Results: The dexamethasone group demonstrated a significantly prolonged sensory block duration (231 +/- 6 vs. 156 +/- 9 minutes; mean difference 75.11 minutes, 95% CI: 71.92-78.29; p < 0.001) and delayed time to first postoperative analgesia (252 +/- 7 vs. 181 +/- 7 minutes; mean difference 71.89 minutes, 95% CI: 68.91-74.86; p < 0.001). Motor block duration was also significantly longer (184 +/- 7 vs. 130 +/- 5 minutes; mean difference 53.42 minutes, 95% CI: 50.99-55.85; p < 0.001). Postoperative pain scores were significantly lower at 1 hour (mean difference -1.29 points, 95% CI: -1.52 to -1.05; p < 0.001) and at 2 hours (mean difference -1.97 points, 95% CI: -2.21 to -1.73; p < 0.001). Intraoperative opioid and benzodiazepine use were significantly reduced in the intervention group. Conclusion: The addition of intrathecal dexamethasone to bupivacaine significantly enhances sensory block duration, delays postoperative analgesia need, and improves early pain control. These findings support its use as a potentially practical adjuvant in resource-limited settings.

BackgroundNon-steroidal anti-inflammatory drugs (NSAIDs) and weak opioids such as tramadol are cornerstones of multimodal analgesia, particu-larly in settings with limited access to potent opioids. However, cross-class equianalgesic data comparing these agents remain scarce. This pilot ran-domised controlled trial aimed to explore the analgesic equivalence of ke-torolac, diclofenac, and tramadol administered as premedication in patients undergoing minimally invasive surgery. MethodsIn this double-blind, parallel-group pilot trial, 30 patients scheduled for elective minimally invasive surgery (28 laparoscopic cholecys-tectomies, 2 laparoscopic abdominal wall repairs) under balanced general anaesthesia were randomised to receive intravenous tramadol 150 mg, ke-torolac 60 mg, or diclofenac 150 mg 45 minutes before skin incision. The primary outcome was pain intensity measured using the Numerical Rating Scale (NRS, 0-10) at recovery room arrival (T0) and at 30 (T1), 60 (T2), and 90 (T3) minutes thereafter. Secondary outcomes included Verbal Rating Scale (VRS) scores, rescue morphine consumption, and safety. Between-group comparisons were performed using Kruskal-Wallis tests with Dunn post-hoc corrections; within-group trajectories were analysed using Fried-man tests. Effect sizes were estimated with epsilon-squared and Kendalls W. ResultsAll 30 patients completed the study. At T0 and T1, NRS scores were higher in the ketorolac group (median 1.5 and 3, respectively) compared with tramadol and diclofenac (both median 0 at T0; T1: tramadol 1, diclofenac 2; p < 0.05 for both). However, by T2 and T3, all three groups converged to a median NRS of 2 (p > 0.05 for between-group differences). Rescue analgesia requirements at T1 were 0/10 (tramadol), 3/10 (ketorolac), and 2/10 (diclofenac), with no statistically significant differences (p = 0.19). No hypersensitivity reactions occurred. Within-group analyses showed con-sistent pain trajectories, with Kendalls W ranging from 0.31 (ketorolac) to 0.64 (tramadol). ConclusionsIn this pilot study, equianalgesic doses of tramadol, ke-torolac, and diclofenac provided comparable postoperative pain control over 90 minutes following minimally invasive surgery. All agents were well toler-ated. These findings support the feasibility of a larger definitive trial and offer clinically useful guidance for analgesic selection in resource-limited settings. Trial registrationClinicalTrials.gov - NCT07500454 (retrospectively registered). HighlightsO_LIDouble-blind pilot RCT compared equianalgesic doses of tramadol, ke-torolac, and diclofenac. C_LIO_LIAll three groups converged to median NRS 2 by 60 minutes postoper-atively. C_LIO_LIEarly higher pain in the ketorolac group was partly attributed to age imbalance ({rho}=0.49, p=0.006). C_LIO_LINo hypersensitivity reactions occurred in any group. C_LIO_LIDefinitive trial requires 27 patients per group (90 total with 10% attri-tion). C_LI

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.

ABSTRACT Background: Self medication with analgesics and non steroidal anti inflammatory drugs (NSAIDs) is common in low- and middle income countries and may expose users to preventable adverse outcomes. Evidence from Guinea remains scarce. This study aimed to estimate the prevalence of self medication with analgesics and NSAIDs among pharmacy clients in urban Conakry, identify associated factors, and describe clinical risk situations. Methods: We conducted a pharmacy based analytical cross sectional study in 30 private pharmacies across Conakry, Guinea. A total of 1,032 participants seeking analgesics or NSAIDs were enrolled between November 3, 2012, and April 5, 2013. Self-medication was defined as acquisition or use without a valid medical prescription. Factors associated with self-medication were analysed using multivariable logistic regression. Results: Among 1,032 participants, 603 reported self medication (prevalence 58.4%). Previous unsupervised use was reported by 78.7%. The most frequently used medicines were paracetamol (56.9%, n=587), diclofenac (21.3%, n=220), ibuprofen (17.9%, n=185), and aspirin (3.9%, n=40). Overall, 68.0% (n=702) reported no knowledge of potential adverse effects. Clinical risk situations were frequent: gastrointestinal disorders (41.3%, n=426), hypertension (9.2%, n=95), and pregnancy exposure among reproductive age women (26.0%). In multivariable analysis, self medication was independently associated with previous analgesic/NSAID use (aOR = 2.8, 95% CI: 2.1 to 3.6), lack of knowledge of adverse effects (aOR = 1.9, 95% CI: 1.4 to 2.5), informal occupation (aOR = 1.6, 95% CI: 1.2 to 2.2), and age 18 to 59 years (aOR = 1.5, 95% CI: 1.1 to 2.1). Conclusions: In this pharmacy based study conducted in urban Conakry, self medication with analgesics and NSAIDs was common and frequently associated with limited awareness of potential adverse effects. These findings support the need for strengthened pharmaceutical regulation, pharmacist-led counselling, health literacy interventions, and improved access to primary care. Keywords: self medication; analgesics; NSAIDs; paracetamol; diclofenac; ibuprofen; pharmacy; Guinea; Conakry; drug safety; public health.

Background and Aims: Synthetic opioids cause tens of thousands of deaths each year in North America, and there are indications that synthetic opioids are also becoming increasingly prevalent in the European drug market. This study aimed to examine high-risk substance use in the German drug-using community with a particular focus on the synthetic opioids fentanyl and nitazenes and related awareness, concerns, overdose experiences, and harm-reduction behavior. Design: Cross-sectional, observational online survey. Setting: Open drug-use scenes, addiction clinics, and substitution practices at numerous geographic locations throughout Germany, August to September 2025. Participants: 235 individuals aged 14+ from the drug using community (mean age 43.4 years; 57.9% male), 79.6% recruited by peers in open drug-use scenes. Measurements: The primary outcome was substances used within the past 12 months. In addition, sources, forms, routes of administration, and perceived changes in availability and price of (synthetic) opioids were assessed as well as risk perceptions, fears, harm-reduction behavior, and overdose-related experiences. Findings: 227 respondents reported substance use with an average of 6.2 substances, and 73.1% (95% confidence interval [CI] = 67.0-78.5%) had used at least one opioid in the past year. Synthetic opioids were consumed in many parts of Germany and across all age and gender groups. Among participants who experienced a shortage of their primary opioid in the past year, 25% (95% CI = 15.8-37.2%) reported having used fentanyl instead. 56.5% (95% CI = 36.8-74.3%) of individuals using synthetic opioids reported having experienced an overdose in the past twelve months. Most of the respondents perceived synthetic opioids as posing a high risk, and a substantial proportion expressed fear that they could be mixed into their own substances. However, only 9.9% (95% CI = 6.6-14.7%) use drug checking, although the vast majority stated they would use it if it were available to them. Conclusions: Synthetic opioids, including fentanyl and nitazenes, have entered the German drug scene, with users reporting high rates of overdose and limited access to harm reduction measures. Germany may be in an early phase of a synthetic opioid transition, warranting urgent expansion of surveillance, naloxone distribution, and drug checking services.

BackgroundThe discontinuation of Fasiglifam (TAK-875), a GPR40/FFAR1 full agonist, during Phase 3 clinical trials due to hepatotoxicity led to widespread abandonment of GPR40 as a viable therapeutic target for type 2 diabetes mellitus (T2DM). However, mechanistic evidence suggests that Fasiglifams hepatotoxicity arises from mitochondrial liability driven by high lipophilicity (aLogP = 5.31), rather than from on-target GPR40 signaling. We hypothesized that target-level failure was incorrectly inferred from compound-level safety concerns, and that superior candidates exist within publicly available databases. MethodsWe queried ChEMBL Release 36 (28 GB SQLite, 74 tables) for all compounds with documented GPR40/FFAR1 activity (UniProt: O14842). Compounds were filtered by EC50 [&le;] 10 nM in nM units with standard relation "=". Drug-likeness was assessed using Lipinskis Rule of Five (Ro5), aLogP, molecular weight (MW), hydrogen bond donors/acceptors (HBD/HBA), and polar surface area (PSA). A parallel analysis of Therapeutic Target Database (TTD v10.1.01, 4,298 targets) provided clinical context. A real-world evidence (RWE) patient stratification framework was constructed using EMR data from tens of millions of patients with >10 years of longitudinal follow-up. ResultsOf 2,637 GPR40-active compounds in ChEMBL 36, 526 (19.9%) demonstrated EC50 < 100 nM and 102 (3.9%) demonstrated EC50 < 10 nM. Eight compounds met stringent drug-likeness criteria (Ro5 violations = 0, aLogP < 5.0, EC50 [&le;] 1 nM). The lead compound (CHEMBL4859651) exhibited EC50 = 0.04 nM (8.75-fold more potent than Fasiglifam), MW = 297 Da (43% lower), and aLogP = 4.30 (19% lower), with zero Ro5 violations. Mean MW of the eight candidates was 317 {+/-} 28 Da versus 524 Da for Fasiglifam. A parallel GCK analysis identified a protein-protein interaction target (CHEMBL3885579, GCK-GKRP interface) harboring 40 exclusive compounds as an orthogonal strategy for partial GCK activation. ConclusionsSystematic cheminformatic analysis reveals that compounds with substantially superior activity and drug-likeness profiles relative to Fasiglifam exist within ChEMBL 36. Fasiglifams hepatotoxicity is attributable to compound-specific physicochemical properties, not GPR40-mediated toxicity. RWE patient stratification may further mitigate hepatotoxicity risk for next-generation GPR40 agonists. These findings argue for systematic reappraisal of GPR40 as a viable therapeutic target for T2DM.

Background: The impact of the Inflation Reduction Act (IRA) upon late-stage developments has been assumed to be limited. The Congressional Budget Office's IRA analysis excluded post-approval innovation, potentially overlooking substantial economic risks to drug developers and declines in the availability of treatments in areas of high unmet medical need such as oncology. Methods: A total of 1148 secondary trials from 364 FDA-approved medicines, published from 2018 to 2025, were obtained from Biomedtracker and clinicaltrials.gov. Using fractional multinomial logit, we model the share distribution of secondary indication studies across 19 disease groups and assess the change in this distribution post-IRA. We also assessed the number of secondary treatment studies pre- vs. post-IRA using multiple linear regression. Results: After the IRA's introduction, small molecule follow-on studies in oncology exhibited a statistically significant 35% decline (R2 = .48, p < 0.014) and lead indication small molecule oncology approvals exhibited a statistically significant 27% decline (R2 = .70, p < 0.002). We also find a statistically significant 14% decline in the share of orphan oncology studies pre- vs. post-IRA (p<0.001). Research Conclusions: This study's results refute claims that the IRA would have minimal negative effects on patient access or late-stage biopharmaceutical R&D. We hope this study reinvigorates debate about the law's unintended consequences and encourages thoughtful policy solutions, as the IRA manifestly creates disincentives that negatively impact patients seeking needed new medicines, particularly those requiring cures addressing metastatic late-stage cancers.

PurposeOpioid overdose deaths disproportionately affect racial and ethnic minority populations in the United States, yet claims-based evidence characterizing the multi-dimensional structure of these disparities across incidence, treatment access, costs, and insurance coverage remains limited. MethodsWe conducted a retrospective cross-sectional and longitudinal cohort analysis using the HealthVerity Launch Sample, a large administrative claims database. The study population comprised 3,675,823 patients across 5 racial groups enrolled between 2020 and 2024. Eight primary analyses were conducted, including age-sex standardized overdose rates, temporal disparity trends, medication-assisted treatment (MAT) receipt, naloxone access, pharmacy costs, insurance payer type, care setting, and multivariable logistic regression for overdose risk. ResultsBlack patients had the highest age-sex standardized overdose rate (363.4 per 100,000; rate ratio [RR] = 1.27 vs. White), and those with opioid use disorder (OUD) received MAT at a rate 35% lower than White patients (19.8% vs. 30.7%; RR = 0.645), driven primarily by a buprenorphine access deficit. AIAN patients demonstrated consistent multi-dimensional disadvantage across naloxone access, MAT engagement, and pharmacy costs. After adjustment for payer type, age, and sex, all non-White groups showed lower adjusted odds of overdose than White patients (Black OR = 0.87; AIAN OR = 0.25), with Medicaid enrollment carrying 7.06 times the overdose odds of commercial insurance. ConclusionInsurance type is the dominant predictor of overdose risk, and the disproportionate Medicaid enrollment of Black patients is both a consequence of structural disadvantage and access disparities. Targeted interventions such as buprenorphine expansion in Medicaid and enhanced naloxone distribution are recommended.

ObjectivesTo evaluate supply-chain vulnerabilities affecting medications essential for treating sexually transmitted infection in the United States and identify disruption mechanisms that may predispose these therapies to shortages. MethodsWe conducted a qualitative, structured supply-chain vulnerability assessment of first-line medications for five priority sexually transmitted pathogens recommended by the Centers for Disease Control and Prevention and the World Health Organization: azithromycin, doxycycline, ceftriaxone, benzathine penicillin G, metronidazole, tinidazole, acyclovir, and cefixime. Using a predefined framework derived from pharmaceutical supply-chain disruption literature, we evaluated 13 disruption categories spanning raw material sourcing, active pharmaceutical ingredient production, manufacturing, distribution, market dynamics, information systems, and post-distribution loss mechanisms. Each category was assessed using four binary indicators and classified as relevant when at least two criteria were satisfied. ResultsMultiple disruption domains applied across the drug set. Recurrent vulnerabilities included geographically concentrated active pharmaceutical ingredient production, limited manufacturing redundancy in low-margin generic markets, manufacturing constraints affecting sterile injectable products, reliance on consolidated distribution networks, and susceptibility to demand surges and information-system disruptions. All eight drugs exhibited at least one regulatory or market signal consistent with potential supply vulnerability, including documented shortages, product discontinuations, or limited manufacturer participation. ConclusionsSupply-chain vulnerabilities were identified across multiple first-line sexually transmitted infection therapies, indicating that disruption risk is not confined to a single drug. There is a need for policy interventions to strengthen supply-chain resilience, including diversification of active pharmaceutical ingredient sourcing and distribution networks, as well as incentives for sustainable generic production.

The rise of new, rapidly mutating viruses presents increasing challenges for drug developers. Traditional methods, such as high-throughput screening and drug repurposing against mutagenic viral targets, have recently shown their limitations. Our current rational molecular engineering approach offers a sustainable solution by targeting viral ion channels, which generally have low mutation rates. First, extending the amantadine molecule led to the development of new compounds that better match the alternating hydrophobic and hydrophilic patterns of the inner walls of ion channels--a common feature across many viruses. Then, simplifying the structure yielded a cyclohexylamine-based minimalist scaffold that effectively blocks the ion channel and demonstrates improved antiviral activity compared to well-known agents such as amantadine and arterolane. SARS-CoV-2 variants served as test systems in laboratory experiments. The new molecular scaffolds presented here provide a strong foundation for designing potent, broad-spectrum viral ion channel blockers.

Ligand optimization is central to drug discovery as hundreds of analogs might be designed and synthesized between an initial hit and a therapeutic candidate. The efficiency of this process is unclear, at least partly because there is no random background for optimization against which to compare. Such a random background might emerge from synthetically accessible but otherwise systematic random small substitutions across starting ligands, measuring likelihood of achieving a substantial improvement in affinity/potency or other property by any single perturbation. Recent literature and ligand-affinity/potency databases suggest that perhaps 10% of analogs with minor modifications improve upon a parents potency substantially (by [&ge;]10-fold), but this number is clouded by reporting bias, intentional improvement, and inter-group reproducibility. To begin to establish a background expectation for ligand optimization, we comprehensively and systematically modified 18 lead molecules across six targets with single atom changes; 257 compounds were synthesized. Unexpectedly, 11.2% of these random small perturbation analogs improved potency by [&ge;]10-fold over their parents. Conversely, these more potent analogs typically had worse in vitro pharmacokinetics (e.g. reduced metabolic stability, lower plasma free fraction). While it was possible to find analogs where the potency increase compensated for inferior exposure and half-life, resulting in more potent compounds in vivo, overall a frustrated landscape for ligand optimization is revealed. This study begins to establish a background expectation for ligand potency optimization and offers a simple strategy to do so. It also begins to quantify the challenges confronting the field in moving beyond in vitro potency.

RationaleFibrotic lung diseases, such as idiopathic pulmonary fibrosis (IPF) and fibroproliferative remodeling in acute respiratory distress syndrome (ARDS), are characterized by increased extracellular matrix (ECM) deposition. However, measuring collagen accumulation alone does not capture differences in ECM organization or biochemical maturation that may distinguish persistent fibrosis from potentially reversible remodeling. ObjectivesTo examine collagen organization characteristics and mature (pyridinoline) collagen crosslinking amount in established end stage fibrotic lung disease (IPF) and fibroproliferation following an acutely damaged lung (non-resolving (NR) ARDS) and to investigate any relationships in these parameters and temporal tissue remodeling. MethodsHuman lung tissue samples from control subjects, patients with IPF, and NR-ARDS were analyzed. Collagen amount and fiber organization were digitally quantified using picrosirius red staining. Mature collagen crosslinking was assessed by quantification of pyridinoline crosslinks. Measurements and Main ResultsLung tissue from both IPF and NR-ARDS lungs had higher collagen content compared with controls. Collagen fiber organization differed between groups. IPF lungs exhibited collagen architectures consistent with established fibrosis, whereas NR-ARDS lungs showed altered but less stabilized collagen organization despite similarly elevated collagen levels. Mature collagen crosslinks were significantly higher in IPF lungs but not in NR-ARDS lungs compared to controls. Integrated analyses identified distinct disease-associated ECM phenotypes, indicating that higher collagen abundance in NR-ARDS, unlike IPF, is not accompanied by more mature and persistent collagen crosslinking. ConclusionsDespite shared increases in collagen content, IPF and NR-ARDS lungs differ fundamentally in collagen organization and crosslinking maturity, suggesting differences in the reversibility of these conditions.

BackgroundCalcitonin gene related peptide (CGRP) hyperpolarizes pulmonary arterial smooth muscle cells (SMCs) and endothelial cells (ECs) through PKA-dependent activation of KATP channels. CGRP can diminish the severity of pulmonary fibrosis (PF), however, the effects on vascular signaling were poorly defined. We hypothesized that hyperpolarization to CGRP would be augmented in a mouse model of PF. MethodsPF was induced in male and female C57BL/6 mice by intratracheal delivery of bleomycin (3 wk), with saline used as control (sham). Pulmonary arteries (PAs; 100-150 {micro}m diameter) were cannulated and pressurized to 16 cmH2O, and endothelial tubes were studied in complementary experiments to eliminate the influence of SMCs. Membrane potential (Vm) was recorded continuously using intracellular microelectrodes. Responses were also evaluated in isolated lungs preconstricted with U46619 ([~]10 mmHg). ResultsPF led to greater indices of PH in males vs. females. Isolated lungs and PAs from male PF mice had enhanced vasodilation and hyperpolarization of Vm to CGRP, although no effect was observed in females. The greater vasodilation and hyperpolarization of SMCs to CGRP in males persisted in endothelium-disrupted PAs and during treatment with L-NAME indicating that ECs are not required for greater responsiveness to CGRP. With no effect on resting Vm, inhibition of KATP channels or PKA significantly attenuated hyperpolarization of SMCs and ECs, attenuated vasodilation to CGRP in PAs, and eliminated differences between groups in males. Direct activation of PKA, but not KATP, evoked greater Vm hyperpolarization and vasodilation in PF vs. sham PAs and lungs. Although no difference in sensory nerves was observed in fibrotic mice, perivascular nerve stimulation evoked greater vasodilation in PAs. ConclusionsIn a mouse model of PF, CGRP-dependent hyperpolarization of pulmonary arterial SMCs and ECs is augmented through increased PKA-dependent activation of KATP channels leading to increased vasodilator sensitivity.

ObjectiveTo 1) determine the expression and distribution of all PDE4 isozymes (A-D) along the length of the anterior urethra, 2) culture fibroblasts and epithelial cells from healthy and strictured urethras, 3) investigate an in vitro model of anterior urethral stricture disease (aUSD), and 4) assess the therapeutic potential of phosphodiesterase-4 (PDE4) inhibitors and testosterone compared to paclitaxel. MethodsThe presence and relative abundance of PDE4 isozymes (A-D) was confirmed using immunohistochemistry on 5 male cadaveric urethras. Human urethral fibroblasts (FBs) were cultured from healthy control urethras of patients undergoing vaginoplasty (n=3) and from idiopathic bulbar urethral strictures (L2S1E2) of patients undergoing urethroplasty (n=3). Epithelial cells (ECs) were cultured from a healthy control urethra and two urethral strictures. To investigate a model of aUSD, Control FBs were stimulated with TGF{beta}1 and compared to Stricture FBs on assays of cell proliferation and expression of genes relevant to aUSD pathophysiology. To test therapeutics, Stricture FBs were treated with the PDE4 inhibitor, roflumilast, testosterone (T), or paclitaxel and compared to Control FBs on the previously mentioned assays and cell viability. ResultsPDE4- A, B, and D were detected along the length of the urethra. Expression levels did not differ between urethral regions. TGF{beta}1 altered proliferation and gene expression in a dose-dependent manner. Roflumilast and T preserved cell viability and proliferation and decreased expression of genes positively associated with auSD. ConclusionUrethral FBs and ECs can be cultured from healthy and strictured surgical specimens, enabling in vitro research. PDE4 inhibitors and T may be non-cytotoxic alternatives or additions to paclitaxel for aUSD. HighlightsO_LIPDE4 isozymes A, B, and D are expressed in adult anterior urethras C_LIO_LIPDE4 is expressed equally from proximal bulbar to meatal urethra C_LIO_LIEpithelial cells and fibroblasts can be cultured from healthy and stricture urethra C_LIO_LITGF{beta}1 may not be an optimal method to model aUSD in vitro C_LIO_LIUnlike paclitaxel, roflumilast and testosterone are not toxic to urethral cells C_LI

The development of lung-directed therapeutics is limited by poor translational fidelity between preclinical models and early-phase clinical trials. We report a first-in-human Phase 0 intratarget microdosing study demonstrating the feasibility of intrapulmonary delivery and pharmacological interrogation of a novel inflammasome inhibitor. A 100 g microdose of ADS032, a dual NLRP1/NLRP3 inhibitor, was administered to distal airways via bronchoscopy in patients with interstitial lung disease, informed by optimisation in ex vivo human lung perfusion and ventilation systems. Clinical-grade manufacture, formulation, stability, and toxicology enabled intrapulmonary administration. Using liquid chromatography-mass spectrometry, ADS032 was detected in plasma, bronchoalveolar lavage fluid, distal airway micro-aspirates, and recovered cells, with spatially resolved sampling achieved without cross-contamination. Fluorescent labelling enabled direct visualisation of alveolar drug uptake ex vivo. These findings establish intrapulmonary intratarget microdosing as a human-relevant platform for early pharmacological evaluation of lung therapeutics prior to Phase 1 trials.

Introduction: Incomplete recovery from relapses contributes to long-term disability accumulation in relapsing remitting multiple sclerosis (RRMS), yet the relationship between immune regulation and relapse recovery remains poorly defined. Objective: To longitudinally characterize regulatory/effector immune cell dynamics in untreated RRMS patients and assess their association with immune balance and relapse recovery. Methods: Monocytic myeloid-derived suppressor cells (M MDSCs), regulatory T cells (Treg), and effector CD4 T cell subsets were measured in blood from 69 untreated RRMS patients sampled during relapse or remission and reevaluated after 12 months. Associations with clinical recovery after relapse were examined. Results: During relapse, patients exhibited higher M MDSC and Treg frequencies than in remission, while effector T cell subsets remained unchanged. Over one year, M-MDSCs increased consistently regardless of baseline clinical status, whereas Treg frequencies remained stable. Effector to M MDSC ratios were markedly elevated during relapse and declined over time, while effector-to-Treg ratios showed minimal variation. M MDSC levels during relapse were associated with sustained regulatory features at 12 month follow up. Importantly, higher baseline M MDSC levels, but not Treg frequencies, were associated with complete relapse recovery at one year. Conclusion: These findings suggest that circulating M-MDSCs, but not Treg, reflect interindividual differences in immune regulation and clinical recovery after relapse in early RRMS.

ObjectivesTo examine the extent to which heat-related causes of death are recorded in fatal drug overdoses, how these patterns vary across states and over time, and how overdose characteristics differ between deaths with, versus without, heat involvement recorded. MethodsDeath certificate data for all drug overdose deaths in US residents from 2001 to 2024 (from the National Center for Health Statistics) were analyzed to identify whether a heat-related cause of death was also listed on the death certificate. Joinpoint regression, descriptive statistics, and nonparametric tests were used to examine temporal trends and compare overdose deaths with versus without recorded heat involvement. ResultsIn 2001, fewer than 10 drug overdose deaths with recorded heat involvement were identified, but this number increased to 558 in 2024. From 2013 to 2024, mortality rates increased significantly, with an estimated annual percent change of 30.1 (95% Confidence Interval, 26.5-47.1). The highest mortality rates and numbers of deaths were observed in residents of Arizona and Nevada. American Indian/Alaska Native, Mexican-heritage, and foreign-born populations accounted for larger shares of overdose deaths with, compared to without, heat involvement recorded. A street or highway was more frequently identified as the place of injury in overdose deaths with (18.9%), versus without (2.2%) heat involvement reported. Psychostimulants such as methamphetamine were involved in 85.9% of overdose deaths with, compared to 28.9% without, recorded heat involvement. ConclusionsAlthough representing only a fraction of all overdose deaths, fatal overdoses involving heat exposure have increased markedly over time and disproportionately impact certain states and demographic groups.

The poor prognosis of brain tumors, including IDH-wild-type glioblastoma (GB), as well as brain and leptomeningeal metastases, is partly related to the blood-brain barrier (BBB), which limits the delivery of hydrophilic anticancer drugs to the tumor site and surrounding brain parenchyma. Early studies using vital dyes demonstrated that intracranial injection could bypass the BBB in cats. We confirmed that, in guinea pigs, the vital dye Bleu Patente V diffused efficiently into the brain after a bolus intracranial injection, whereas the brain remained unstained after intravenous administration. Similarly, brain concentrations of the hydrophilic anticancer drug gemcitabine were significantly higher following intracranial injection than after intravenous administration. Consistent with these findings, Bleu Patente penetrated deeply into the cerebral cortex of sheep after a 24-hour intraventricular infusion. At the end of a 24-hour intraventricular infusion of 20 mg gemcitabine in sheep, mean gemcitabine concentrations reached 1,415 {micro}g/L in cerebrospinal fluid and 850 {micro}g/kg in brain tissue. These concentrations exceeded the IC90 values of gemcitabine for A172, U87-MG, and U118-MG human glioblastoma cell lines, as determined in vitro after 24 hours of incubation. We hypothesize that Bleu Patente dye and gemcitabine circumvent the blood-brain barrier (BBB) by utilizing the glymphatic system. Tolerance of a single 24-hour intraventricular infusion of gemcitabine at doses of 5, 10, and 20 mg was good. Taken together, these encouraging preclinical results support the resumption of Phase I clinical trials evaluating intraventricular infusion of gemcitabine in patients with refractory primary or secondary brain tumors.