Search Results (110)

Background and Objectives: A considerable proportion of patients with asthma remain inadequately controlled despite treatment with inhaled corticosteroid/long-acting β₂-agonist (ICS/LABA) therapy. Single-inhaler triple therapy comprising an inhaled corticosteroid, a long-acting β₂-agonist, and a long-acting muscarinic antagonist (ICS/LABA/LAMA) has emerged as an effective step-up treatment; however, real-world evidence remains limited. This study aimed to evaluate the effects of transitioning from high-dose ICS/LABA therapy to single-inhaler triple therapy on asthma control, lung function, and exacerbation-related outcomes in routine clinical practice. Materials and Methods: This retrospective observational study included adult patients with uncontrolled asthma who were switched from high-dose ICS/LABA therapy to single-inhaler triple therapy at a tertiary referral center. Demographic characteristics, Asthma Control Test (ACT) scores, pulmonary function tests, blood eosinophil counts, exacerbation frequency, oral corticosteroid (OCS)-requiring exacerbations, short-acting β₂-agonist (SABA) use, emergency department visits, and hospitalizations were compared before and after treatment escalation using paired statistical analyses. Results: Nineteen patients (mean age 58.9 ± 10.4 years; 63.2% female) were included. Following transition to single-inhaler triple therapy, asthma control improved significantly, with mean ACT scores increasing from 14.8 ± 3.2 to 23.4 ± 3.6 (p < 0.001). Mean forced expiratory volume in one second (FEV₁₎ increased from 1692.6 ± 722.7 mL to 2020.5 ± 871.5 mL, while predicted FEV₁ improved from 64.2 ± 21.2% to 75.6 ± 21.6% (both p < 0.001). Total annual exacerbation frequency decreased from a median of 2 (interquartile range [IQR], 1–3) to 0 (IQR, 0–1; p = 0.0005), and 78.9% of patients experienced at least one fewer exacerbation. Significant reductions were also observed in OCS-requiring exacerbations, SABA use, emergency department visits, hospitalizations, and blood eosinophil counts. Conclusions: In this real-world cohort, transitioning from ICS/LABA therapy to single-inhaler triple therapy was associated with statistically significant improvements in asthma control and lung function, together with substantial reductions in exacerbation burden and healthcare utilization. These findings support the effectiveness of single-inhaler triple therapy as a step-up treatment strategy for patients with uncontrolled asthma in routine clinical practice. Full article

Cantharidin (CANT) and sodium fluoride inhibit serine/threonine phosphatase (PP) 1 and PP2A in the heart. We hypothesized that Cant and sodium fluoride could weaken the reduction in force by carbachol in the presence of serotonin in human right atrial preparations (HAP). We measured contractility in HAP, and for comparison, in left atria from wild type mice (CD1, WT), mice with human 5-HT₄-serotonin receptor cardiac overexpression (5-HT₄-TG), or in mice with human H₂-histamine receptor cardiac overexpression (H₂-TG). Isoprenaline (1 µM), histamine (1 µM), and serotonin (1 µM) raised contraction in HAP. Likewise, isoprenaline (1 µM), histamine (1 µM), and serotonin (1 µM) raised contractility in the left atria of WT, H₂-TG, and 5-HT₄-TG. Carbachol (1 µM), a muscarinic cholinoceptor agonist, diminished the force of contraction in a time-dependent manner after prior stimulation of force by serotonin, isoprenaline, or histamine in HAP but also in left atria from 5-HT₄-TG, WT, and H₂-TG. These anti-β-adrenergic, anti-serotoninergic and anti-histaminergic effects of carbachol on force of contraction were attenuated by 100 µM or 30 µM cantharidin or 3 mM sodium fluoride in HAP and in left atrial preparations from WT, H₂-TG, and 5-HT₄-TG, respectively. We conclude that muscarinic cholinoceptor activation may exert a negative inotropic effect in the HAP by stimulating PP1 and/or PP2A, which had previously been inhibited by isoprenaline or histamine or serotonin. Full article

Background: Bladder dysfunction is a complicated condition that substantially impairs quality of life for both men and women. Due to the adverse effects and limited efficacy of current therapies, new strategies must be rapidly developed. Female bladder dysfunction arises from multifaceted etiologies distinct from the predominantly male benign prostatic hyperplasia (BPH) that is the focus of existing drug development. In this study, we investigated the therapeutic potential of Quisqualis indica extract (QIE), a traditional medicinal herb that attenuates BPH-induced lower urinary symptoms (LUTS), to elucidate its underlying mechanisms in a female bladder dysfunction model. Methods and Results: A bladder dysfunction model was established by inducing partial bladder outlet obstruction (pBOO) in female Sprague Dawley rats, followed by the oral administration of QIE for 7 weeks. Voiding pattern analysis and cystometry were conducted to evaluate indicators such as voiding frequency, voiding volume, and intravesical pressure. Histological analysis of excised bladder tissue quantified smooth muscle hypertrophy and collagen deposition. Gene expression profiling of inflammatory cytokines and fibrosis-related markers within the bladder tissue was performed to assess tissue remodeling. Furthermore, pharmacological contraction studies examined the direct effects of QIE on detrusor muscle responsiveness to muscarinic and purinergic agonists. QIE administration significantly improved the elevated voiding pressure and abnormal inter-contraction intervals observed in the pBOO rats, restoring normal voiding patterns. Histological examination revealed a marked decrease in muscle hypertrophy and collagen deposition. Expression levels of pro-inflammatory cytokines (TNFα, IL-1β) and fibrosis-associated genes (TGF-β, α-SMA) were downregulated. Pharmacological contraction assays demonstrated that QIE attenuated the hypercontractile response of bladder smooth muscle to a muscarinic agonist, with concurrent reduced expression of muscarinic receptors (M₂, M₃) at the mRNA level. Conclusions:QIE ameliorates key aspects of bladder dysfunction, voiding abnormalities, inflammation, fibrosis, and hypercontractility by modulating muscarinic receptor signaling and fibrotic pathways. This study suggests that QIE warrants further investigation as a natural product-based therapeutic candidate for female bladder dysfunction. Full article

Little is known about how nicotinic signalling in human astrocytes may contribute to the functional neurotoxicity of compounds related to tobacco alkaloids and neonicotinoid pesticides. We generated a single-cell Ca²⁺-imaging assay in induced pluripotent stem cell (iPSC)-derived astrocytes, and profiled functional expressions of some neurotoxicologically relevant receptors. Responses to pharmacological tool compounds indicated the expression of nicotinic, muscarinic, purinergic, glutamatergic receptors and voltage-gated Na⁺/Ca²⁺ channels. Closer investigation of the nicotinic system, e.g., using the alpha7 nicotinic acetylcholine receptor (nAChR)-selective positive allosteric modulator PNU-120596 and alpha7-preferring agonist (AR-R17779) demonstrated that Ca²⁺ signals elicited by nicotine and neonicotinoids are dominated by alpha7 nAChRs and depend on the downstream activation of L-type Ca²⁺ channels and tetrodotoxin-sensitive Na⁺ channels. Crosstalk of nAChR activation/desensitization was not observed for the inflammatory response elicited by TNF or for activation of glutamatergic or purinergic signalling. However, pre-stimulation of nAChR by neonicotinoids significantly blunted the response to the neurotransmitter acetylcholine. Comparative experiments in the human neuronal cultures (LUHMES cells) revealed similar potency ranges and pharmacological fingerprints for several neonicotinoids and their human-relevant metabolites descyanothiacloprid and desnitroimidacloprid. The pesticide metabolites showed a high potency, compared with their respective parent compounds. After this basic system characterization, the hitherto data-poor pesticides cycloxaprid and flupyradifurone were comparatively profiled in astrocytic and neuronal test systems. They showed the typical features of alpha7 nAChR agonists. The disruption of cholinergic signalling in astrocytes suggests that neonicotinoids affect not only neurons in human brains. Therefore, future neurotoxicity screening approaches may need to consider astrocyte toxicity. Full article

Schizophrenia remains a debilitating global health challenge where pharmacologic treatment has been stagnant for over seventy years, relying almost exclusively on the blockade of dopamine receptors. While this mechanism controls positive psychosis, it frequently fails to address negative symptoms or cognitive impairment and carries a significant burden of metabolic and motor adverse effects. This review evaluates the scientific and clinical validation of xanomeline–trospium (Cobenfy^®; investigational name KarXT), the first approved antipsychotic with a completely non-dopaminergic mechanism of action. We synthesize data ranging from the unique structural biology of the bitopic M1/M4 muscarinic receptor agonist xanomeline to the pharmacokinetic innovation of using the peripheral antagonist trospium chloride to mitigate systemic toxicity. Comprehensive analyses of the EMERGENT clinical trial program demonstrate that this combination significantly reduces heterogeneous schizophrenia symptoms with a safety profile distinct from current standards of care, specifically avoiding weight gain and extrapyramidal movement disorders. Furthermore, we contrast this success with the recent failures of other novel mechanisms and explore the potential for precision medicine through the identification of muscarinic receptor deficit biotypes. We conclude that M1/M4 muscarinic receptor agonism represents an important advance toward circuit-based therapeutics that may help overcome some of the limitations inherent to dopamine-centered pharmacotherapy. Full article

Background: Overuse of short-acting reliever inhalers, particularly short-acting β₂-agonists (SABAs), is linked to poor asthma control, higher exacerbation risk, and increased mortality. Data on reliever overuse in severe asthma and the role of short-acting muscarinic antagonists (SAMAs) remain limited. Objective: This study aims to assess the prevalence of rescue inhaler overuse in adults with severe asthma and examine its associations with maintenance therapy adherence, biologic treatment, oral corticosteroid use, asthma control, lung function, allergic sensitization, and comorbidities. Methods: We conducted a retrospective observational study of 223 adults with severe asthma followed at a tertiary multidisciplinary clinic in 2024. Clinical, functional, pharmacological, and pharmacy-dispensing records were reviewed. Rescue inhaler overuse was defined as dispensing ≥ 3 SABA and/or SAMA canisters per 12 months. Associations with clinical and treatment variables were analyzed. Results: Among 223 patients, 144 (64.6%) had a prescribed rescue inhaler; 85 (38.1%) met the criteria for overuse. Of those prescribed rescue therapy, 59.0% overused, with 47.2% classified as low overuse and 11.1% as high overuse. Overuse was more frequent in patients with maintenance adherence > 50%. Biologic therapy was associated with reduced odds of overuse, while oral corticosteroid use showed no significant effect. Poor asthma control (ACT < 20) was strongly associated with overuse. Lung function did not differ significantly, though overusers tended to have lower FEV₁. Allergic sensitization was not associated with overuse; bronchiectasis was the comorbidity most frequently linked. Conclusions: Rescue inhaler overuse is common in severe asthma and closely linked to inadequate disease control. Lower overuse rates among biologic-treated patients suggest improved control reduces reliance on short-acting relievers. Systematic monitoring of reliever use may help identify high-risk patients and guide individualized management strategies. Full article

Background/Objectives: Inflammatory bowel diseases (IBD) are conditions of the gastrointestinal tract with treatments linked to side effects and relapses. Spondias mombin L. is a species with anti-inflammatory action, but there is little information in the literature about its application in the treatment of IBD. The aim of this study was to evaluate the intestinal anti-inflammatory activity of hydroalcoholic extract of Spondias mombin L. (HESm) in a rat model of ulcerative colitis (UC). Methods: Hydroalcoholic extract was obtained using the turboextraction technique, followed by identification of its major components using ultra-high performance liquid chromatography (UFLC). Intestinal anti-inflammatory activity was evaluated in an acute model of UC induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Groups received a vehicle, prednisolone (2 mg/kg), or HESm (125, 250 or 500 mg/kg) before and after UC induction. Ulcerated area, score, and intestinal weight/length ratio were analyzed. Histopathological analysis and the extract’s effect on the contractility of the intestinal segment were carried out. Results: UFLC identified the presence of quercetin, a flavonoid widely cited for the species. At doses of 125, 250, and 500 mg/kg, the extract reduced areas of injury by 86.82, 92.67, and 85.06%, respectively, compared to the control, in addition to reducing scores and weight/length ratio of the colons. Histopathological analysis confirmed the results. In contractility, the extract at the highest concentration tested reduced the response of the muscarinic agonist carbamylcholine to 53.7 ± 4.2% of control contraction. Conclusions: Results demonstrate the species’ ability to reduce injuries caused by colitis, suggesting its potential to contribute to the clinical management of IBD in the future. Full article

Background: The discovery of electroencephalogram (EEG) biomarkers of direct transmitter–receptor interactions in studies of neurotransmitter mechanisms underlying brain function remains relevant. Recently, EEG “signatures” of monoaminergic systems have been demonstrated using the “time-frequency clustering” approach. In the current study, the glutamic and cholinergic systems were under similar analysis with additional emphasis on their potential interaction. Methods: In non-anesthetized freely moving rats, we studied the EEG effects of agonists for glutamate receptors, injected into the cerebral lateral ventricles, and their modification after pretreatment with corresponding antagonists. The same protocol was used for acetylcholine receptors, activating and blocking substances that penetrate the blood–brain barrier (BBB) after subcutaneous injections. A clustering of significant time-dependent changes in tiny frequency subranges of the EEG spectra was performed. Results: After injections of agonists for glutamate receptors, two clusters with enhanced and suppressed activities around 2/4 and 10 Hz, respectively, were observed in the EEG spectra. These effects were reduced by pretreatment with corresponding receptor blockers. A cholinomimetic, physostigmine, decreased EEG activity around 2 and 10 Hz and increased near 5 and 22 Hz. Scopolamine, blocking muscarinic cholinoreceptors, weakened the effects of physostigmine. Intracerebral pretreatment with NMDA and AMPA receptor blockers differently modified the effects of physostigmine. The results demonstrate the EEG biomarkers of glutamatergic and cholinergic systems, as well as the specificity of interactions between them at the intracerebral level. Conclusions: The developed EEG time-frequency clustering is a potentially useful approach for the clinical evaluation of glutamatergic/cholinergic pathology and its correction by corresponding substances penetrating the BBB. Full article

The role of cholinergic system in hair biology is poorly understood. In M4 muscarinic receptor (mAChR) knockout mice, the hair follicles have a prolonged telogen phase and fail to produce hair shafts. Here, we reported that hair growth was regulated by cholinergic signalling via mAChRs. Dermal papilla cells expressed different cholinergic biomarkers. Inhibiting AChE or activating mAChR in dermal papilla cells, cultured vibrissae and mouse skin epidermis promoted the hair growth. In cultured papilla cells treated with bethanechol, an agonist of mAChR, an activation of Wnt/β-catenin signalling was illustrated by various indicative biomarkers, including phosphorylation of GSK-3β and mRNA expression of various molecules for Wnt/β-catenin signalling. Activation of Wnt/β-catenin signalling was mediated by PI3K/AKT and ERK signalling upon the stimulation of bethanechol. Moreover, the bethanechol-mediated signalling was blocked by specific antagonist of M4 mAChR. In addition, an increase in hair shaft elongation was observed in mouse vibrissae upon the treatment of bethanechol, suggesting the cholinergic role in hair growth. Full article

Older adults with multiple diseases are likely to be prescribed multiple medications including anticholinergic agents, which are frequently prescribed to manage conditions such as overactive bladder and chronic obstructive pulmonary disease and Parkinson’s disease. Overactive bladder (OAB) has been the subject of increased disease awareness and is a common and significant cause of reduced quality of life, particularly in the elderly. The selective β₃ adrenoceptor agonist, mirabegron was developed for the pharmacological treatment of OAB. Mirabegron has been shown to exert off-target effects on various functional proteins such as muscarinic receptors in rat tissues. This agent may relax the detrusor muscle by activating β₃ adrenoceptors and also antagonizing muscarinic receptors. Mirabegron and antimuscarinics exerted additive effects on muscarinic receptor binding and relaxant responses of cholinergic contractions of the detrusor muscle. Mirabegron excreted in human urine appears to directly attenuate muscarinic receptor-mediated functions in the bladder. Combination therapy of mirabegron and solifenacin in patients with OAB may enhance not only their therapeutic effects on OAB, but also increase the risk of anticholinergic adverse effects. Therefore, the safety of concomitant use of mirabegron and other drugs such as antimuscarinics for elderly patients needs to be carefully considered. Full article

To better address the challenge of corneal ulcer healing, with already available standard agents, and those recently introduced, such as stable gastric pentadecapeptide BPC 157, we introduced a novel conceptual framework—the “triad” of corneal ulcer healing↔corneal neovascularization↔intraocular pressure—and extended it to avascular tissues such as tendon. Within this framework, cytoprotection serves as the unifying principle, underscoring that therapeutic effects are not isolated but interconnected. Preclinical studies with BPC 157 therapy, as a cytoprotection agent, illustrate this integration. BPC 157 rapidly normalizes elevated intraocular pressure in glaucomatous rats, preserves retinal integrity, restores pupil function, maintains corneal transparency during ulcer or abrasion healing, and counteracts both corneal neovascularization and dry eye. In parallel, its consistent efficacy in tendon injury models highlights a cytoprotective specificity across avascular tissues. The cornea’s “angiogenic privilege,” preserved during healing and tendon recovery together, provides strong proof of concept. Furthermore, mapping standard therapeutic agents used for corneal ulcers, neovascularization, or glaucoma onto this triad, and linking them with tendon healing, reveals both shared pathways and inconsistencies across existing drug classes. Analyzed were the ascorbate, fibronectin, hyaluronic acid, metalloproteinase inhibitors, EGF, FGF, NGF, insulin, and IGF-1 (corneal ulcer healing), the antiangiogenic agents (endostatin, PAI-1, PEDF, angiostatin, TSP-1, TSP-2, IFN-α), corticosteroids, NSAIDs, cyclosporine A, anti-VEGF drops (treatment of corneal neovascularization), and alpha 2-agonists, beta-blockers, carboanhydrase inhibitors, muscarinic agonists, Rho-kinase inhibitors, and prostaglandin analogs (glaucoma). Taken together, these findings advance cytoprotection as a unifying therapeutic paradigm, with BPC 157 emerging as its first exemplar, and encourage further translational research toward clinical application. Full article

Inhibition of agonist-induced M₂ muscarinic receptor (M₂R) activation by functional anti-M₂R autoantibodies has been associated with cardiac parasympathetic dysfunction in patients with chronic Chagas disease (CD). This study explored the allosteric nature of that inhibitory effect by assessing the ability of serum IgG from patients with CD and dysautonomia (DCD IgG) to modulate the interaction between M₂R and β-arrestins in HEK 293T cells using bioluminescence resonance energy transfer. DCD IgG alone did not stimulate arrestin-2 or arrestin-3 recruitment. When cells were preincubated with DCD IgG and then treated with carbachol, arrestin-2 translocation decreased in a concentration-dependent manner, while arrestin-3 recruitment remained unaffected. Inhibition curve analysis showed a submaximal inhibitory effect (68.1 ± 2.4%) and a Hill slope less than −1 (−4.03 ± 0.39). Carbachol concentration–response assays after preincubation with DCD IgG revealed a noncompetitive inhibition of arrestin-2 recruitment, with no change in arrestin-3 translocation. Unlikely, simultaneous exposure to DCD IgG and carbachol potentiated agonist-induced Arr-2 recruitment. We conclude that anti-M₂R autoantibodies selectively inhibit agonist-induced arrestin-2 recruitment, acting as negative allosteric modulators of agonist efficacy. The direction of autoantibody-induced allosteric modulation depends on the timing of IgG application relative to the agonist and the duration of receptor exposure to autoantibodies. Full article

Teleost fish are the first evolutionary group to exhibit an innate and adaptive immune system. Within the mechanisms of adaptive immunity, fish possess, among others, T-helper cells (CD4-like) and their differentiation machinery, regulated by the master transcription factors T-bet, GATA3, Foxp3, and RORγ. Many studies support the existence of a non-neuronal cholinergic system involved in the immune response, named after the ability of leukocytes to synthesize de novo acetylcholine (ACh). Organophosphorus pesticides (OPs), such as diazoxon (DXN), are examples of compounds that act as cholinergic disruptors with immunotoxic effects. The present study aimed to evaluate the expression of transcription factors in leukocytes (spleen mononuclear cells, SMNCs) of Nile tilapia by modulating cholinergic pathways in immune cells using agonists, antagonists, and diazoxon (DXN), an anticholinesterase substance. The obtained data showed a significant increase in RORγ mRNA expression upon stimulation with the nicotinic agonist, whereas activation of the muscarinic receptor with its agonist increased T-bet mRNA expression. An alteration in RORγ expression levels induced by DXN exposure was also observed. The results suggest a probable directing of the immune response towards a pro-inflammatory profile orchestrated mainly by RORγ and T-bet transcription factors in response to cholinergic stimuli. Full article

In chronic obstructive pulmonary disease (COPD), dysregulated calcium homeostasis, oxidative stress, and mucus hypersecretion converge to suppress ciliary beat frequency (CBF), thereby compromising mucociliary clearance (MCC). These mechanisms are subject to pharmacological modulation. Long-acting muscarinic antagonists (LAMAs) exert direct cilia-stimulatory effects and may counteract pathogen-induced mucin overproduction without impairing clearance. Long-acting β₂-agonists (LABAs) enhance ciliary activity through the cAMP–PKA–dynein (cyclic adenosine monophosphate–protein kinase A–dynein) signalling pathway. Inhaled corticosteroids (ICSs), although largely neutral on CBF, provide indirect protection by suppressing IL-13–driven inflammation. Phosphodiesterase (PDE)-4 inhibitors sustain intracellular cAMP and promote ciliary motility, though their clinical use remains limited by adverse effects. Emerging evidence suggests that dual and triple therapies may provide additive or synergistic benefits for preserving mucociliary function. Clinically, ex vivo CBF interpretation may be influenced by ongoing pharmacotherapy and tissue sampling site. Nasal brush samples may predominantly reflect systemic rather than inhaled therapy. Moreover, differences in PDE isoform expression between nasal and bronchial epithelium further complicate direct extrapolation of results. Rigorous patient stratification by treatment regimen is therefore essential to reconcile inconsistencies reported across studies. Ultimately, preservation of MCC in COPD depends on a delicate balance between oxidative stress and pharmacological modulation of ciliary function. Full article

Background: Triple therapy (long-acting muscarinic antagonists (LAMAs), long-acting beta agonists (LABAs) and inhaled corticosteroids (ICSs)) is a recommended treatment for moderate-to-severe asthma at GINA Steps 4 and 5. However, little is known about the acceptance and use of triple therapy in everyday practice. The EU-LAMA Survey assessed specialists’ knowledge and views on triple therapy in daily practice. Methods: A 19-question survey was administered to 630 pulmonologists, allergologists, general practitioners, and internal medicine specialists in Poland (58%), Greece (27%), Sweden (6.3%), Slovenia (5.4%), and Austria (3.7%) using a dedicated online platform and computer-assisted web interviews. Results: The majority of the physicians were pulmonologists (59%), followed by allergologists (15.7%). For uncontrolled asthma at GINA Step 4, 81% of the respondents preferred increasing the ICS dose to the maximum level, whereas 76% opted to add LAMAs to medium-dose ICSs. At GINA Step 5, 79% of the respondents chose LAMAs first, followed by biological therapy (51%). Oral corticosteroids were favored over increasing the ICS dose and adding LAMAs. Triple therapy was mostly administered in one inhaler (70% and 82% at GINA Steps 4 and 5, respectively). Barriers to the use of LAMAs included a lack of reimbursement (31%), unclear guidelines (24%), lack of experience (18%), insufficient evidence (13%), fear of step-up regimens (10%), and the ease of increasing ICS doses (9%). Conclusion: Many physicians continue to rely on oral corticosteroids at GINA Steps 4 and 5 and infrequently refer patients to triple therapy or biological treatments at GINA Step 5. Full article