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Prostaglandin E2 imparts diverse physiological effects on multiple airway cells through its actions on four distinct E-type prostanoid (EP) receptor subtypes (EP1-EP4). Gs-coupled EP2 and EP4 receptors are expressed on airway smooth muscle (ASM), yet their capacity to regulate the ASM contractile state remains subject to debate. We used EP2 and EP4 subtype-specific agonists (ONO-259 and ONO-329, respectively) in cell- and tissue-based models of human ASM contraction--magnetic twisting cytometry (MTC), and precision-cut lung slices (PCLSs), respectively--to study the EP2 and EP4 regulation of ASM contraction and signaling under conditions of histamine or methacholine (MCh) stimulation. ONO-329 was superior (<0.05) to ONO-259 in relaxing MCh-contracted PCLSs (log half maximal effective concentration [logEC50]: 4.9 x 10-7 vs. 2.2 x 10-6; maximal bronchodilation ± SE, 35 ± 2% vs. 15 ± 2%). However, ONO-259 and ONO-329 were similarly efficacious in relaxing histamine-contracted PCLSs. Similar differential effects were observed in MTC studies. Signaling analyses revealed only modest differences in ONO-329- and ONO-259-induced phosphorylation of the protein kinase A substrates VASP and HSP20, with concomitant stimulation with MCh or histamine. Conversely, ONO-259 failed to inhibit MCh-induced phosphorylation of the regulatory myosin light chain (pMLC20) and the F-actin/G-actin ratio (F/G-actin ratio) while effectively inhibiting their induction by histamine. ONO-329 was effective in reversing induced pMLC20 and the F/G-actin ratio with both MCh and histamine. Thus, the contractile-agonist-dependent differential effects are not explained by changes in the global levels of phosphorylated protein kinase A substrates but are reflected in the regulation of pMLC20 (cross-bridge cycling) and F/G-actin ratio (actin cytoskeleton integrity, force transmission), implicating a role for compartmentalized signaling involving muscarinic, histamine, and EP receptor subtypes. [ABSTRACT FROM AUTHOR]

Rhinoviruses (RVs) evoke as many as 85% of acute asthma exacerbations in children and 50% in adults and can induce airway hyperresponsiveness and decrease efficacy of current therapeutics to provide symptom relief. Using human precision-cut lung slices (hPCLSs), primary human air–liquid interface–differentiated airway epithelial cells (HAECs), and human airway smooth muscle (HASM) as preclinical experimental models, we demonstrated that RV-C15 attenuates agonist-induced bronchodilation. Specifically, airway relaxation to formoterol and cholera toxin, but not forskolin (Fsk), was attenuated following hPCLS exposure to RV-C15. In isolated HASM cells, exposure to conditioned media from RV-exposed HAECs decreased cellular relaxation in response to isoproterenol and prostaglandin E2, but not Fsk. Additionally, cAMP generation elicited by formoterol and isoproterenol, but not Fsk, was attenuated following HASM exposure to RV-C15–conditioned HAEC media. HASM exposure to RV-C15–conditioned HAEC media modulated expression of components of relaxation pathways, specifically GNAI1 and GRK2. Strikingly, similar to exposure to intact RV-C15, hPCLS exposed to UV-inactivated RV-C15 showed markedly attenuated airway relaxation in response to formoterol, suggesting that the mechanism(s) of RV-C15–mediated loss of bronchodilation is independent of virus replication pathways. Further studies are warranted to identify soluble factor(s) regulating the epithelial-driven smooth muscle loss of β2-adrenergic receptor function. [ABSTRACT FROM AUTHOR]

Although metastasis is the leading cause of cancer deaths, it is quite rare at the cellular level. Only a rare subset of cancer cells (~ 1 in 1.5 billion) can complete the entire metastatic cascade: invasion, intravasation, survival in the circulation, extravasation, and colonization (i.e. are metastasis competent). We propose that cells engaging a Polyaneuploid Cancer Cell (PACC) phenotype are metastasis competent. Cells in the PACC state are enlarged, endocycling (i.e. non-dividing) cells with increased genomic content that form in response to stress. Single-cell tracking using time lapse microscopy reveals that PACC state cells have increased motility. Additionally, cells in the PACC state exhibit increased capacity for environment-sensing and directional migration in chemotactic environments, predicting successful invasion. Magnetic Twisting Cytometry and Atomic Force Microscopy reveal that cells in the PACC state display hyper-elastic properties like increased peripheral deformability and maintained peri-nuclear cortical integrity that predict successful intravasation and extravasation. Furthermore, four orthogonal methods reveal that cells in the PACC state have increased expression of vimentin, a hyper-elastic biomolecule known to modulate biomechanical properties and induce mesenchymal-like motility. Taken together, these data indicate that cells in the PACC state have increased metastatic potential and are worthy of further in vivo analysis. [ABSTRACT FROM AUTHOR]

Background and Purpose: β-Adrenoceptor agonists relieve airflow obstruction by activating β2 -adrenoceptors, which are G protein-coupled receptors (GPCRs) expressed on human airway smooth muscle (HASM) cells. The currently available β-adrenoceptor agonists are balanced agonists, however, and signal through both the stimulatory G protein (Gs )- and β-arrestin-mediated pathways. While Gs signalling is beneficial and promotes HASM relaxation, β-arrestin activation is associated with reduced Gs efficacy. In this context, biased ligands that selectively promote β2 -adrenoceptor coupling to Gs signalling represent a promising strategy to treat asthma. Here, we examined several β-adrenoceptor agonists to identify Gs -biased ligands devoid of β-arrestin-mediated effects.Experimental Approach: Gs -biased ligands for the β2 -adrenoceptor were identified by high-throughput screening and then evaluated for Gs interaction, Gi interaction, cAMP production, β-arrestin interaction, GPCR kinase (GRK) phosphorylation of the receptor, receptor trafficking, ERK activation, and functional desensitization of the β2 -adrenoceptor.Key Results: We identified ractopamine, dobutamine, and higenamine as Gs -biased agonists that activate the Gs /cAMP pathway upon β2 -adrenoceptor stimulation while showing minimal Gi or β-arrestin interaction. Furthermore, these compounds did not induce any receptor trafficking and had reduced GRK5-mediated phosphorylation of the β2 -adrenoceptor. Finally, we observed minimal physiological desensitization of the β2 -adrenoceptor in primary HASM cells upon treatment with biased agonists.Conclusion and Implications: Our work demonstrates that Gs -biased signalling through the β2 -adrenoceptor may prove to be an effective strategy to promote HASM relaxation in the treatment of asthma. Such biased compounds may also be useful in identifying the molecular mechanisms that determine biased signalling and in design of safer drugs. [ABSTRACT FROM AUTHOR]

G protein-coupled receptors display multifunctional signaling, offering the potential for agonist structures to promote conformational selectivity for biased outputs. For β2-adrenergic receptors (β2AR), unbiased agonists stabilize conformation(s) that evoke coupling to Gαs (cyclic adenosine monophosphate [cAMP] production/human airway smooth muscle [HASM] cell relaxation) and β-arrestin engagement, the latter acting to quench Gαs signaling, contributing to receptor desensitization/tachyphylaxis. We screened a 40-million-compound scaffold ranking library, revealing unanticipated agonists with dihydroimidazolyl-butylcyclic urea scaffolds. The S-stereoisomer of compound C1 shows no detectable β-arrestin engagement/signaling by four methods. However, C1-S retained Gαs signaling--a divergence of the outputs favorable for treating asthma. Functional studies with two models confirmed the biasing: β2AR-mediated cAMP signaling underwent desensitization to the unbiased agonist albuterol but not to C1-S, and desensitization of HASM cell relaxation was observed with albuterol but not with C1-S. These HASM results indicate biologically pertinent biasing of C1-S, in the context of the relevant physiologic response, in the human cell type of interest. Thus, C1-S was apparently strongly biased away from β-arrestin, in contrast to albuterol and C5-S. C1-S structural modeling and simulations revealed binding differences compared with unbiased epinephrine at transmembrane (TM) segments 3,5,6,7 and ECL2. C1-S (R2 = cyclohexane) was repositioned in the pocket such that it lost a TM6 interaction and gained a TM7 interaction compared with the analogous unbiased C5-S (R2 = benzene group), which appears to contribute to C1-S biasing away from β-arrestin. Thus, an agnostic large chemical-space library identified agonists with receptor interactions that resulted in relevant signal splitting of β2AR actions favorable for treating obstructive lung disease. [ABSTRACT FROM AUTHOR]

Management and regulatory agencies face a wide range of environmental issues globally. The challenge is to identify and select the issues to assist the allocation of research and policy resources to achieve maximum environmental gain. A framework was developed to prioritize environmental contamination issues in a sustainable management policy context using a nine-factor ranking model to rank the significance of di use sources of stressors. It focuses on contamination issues that involve large geographic scales (e.g., all pastoral soils), significant population exposures (e.g., urban air quality), and multiple outputs from same source on receiving environmental compartments comprising air, surface water, groundwater, and sediment. Factor scores are allocated using a scoring scale and weighted following defined rules. Results are ranked enabling the rational comparison of dissimilar and complex issues. Advantages of this model include flexibility, transparency, ability to prioritize new issues as they arise, and ability to identify which issues are comparatively trivial and which present a more serious challenge to sustainability policy goals. This model integrates well as a planning tool and has been used to inform regional policy development. [ABSTRACT FROM AUTHOR]

The recent discovery of sensory (tastant and odorant) G proteincoupled receptors on the smooth muscle of human bronchi suggests unappreciated therapeutic targets in the management of obstructive lung diseases. Here we have characterized the effects of a wide range of volatile odorants on the contractile state of airway smooth muscle (ASM) and uncovered a complex mechanism of odorant-evoked signaling properties that regulate excitation-contraction (E-C) coupling in human ASM cells. Initial studies established multiple odorous molecules capable of increasing intracellular calcium ([Ca2+]i) in ASM cells, some of which were (paradoxically) associated with ASM relaxation. Subsequent studies showed a terpenoid molecule (nerol)-stimulated OR2W3 caused increases in [Ca2+]i and relaxation of ASM cells. Of note, OR2W3-evoked [Ca2+]i mobilization and ASM relaxation required Ca2+ flux through the store-operated calcium entry (SOCE) pathway and accompanied plasma membrane depolarization. This chemosensory odorant receptor response was not mediated by adenylyl cyclase (AC)/cyclic nucleotide-gated (CNG) channels or by protein kinase A (PKA) activity. Instead, ASM olfactory responses to the monoterpene nerol were predominated by the activity of Ca2+-activated chloride channels (TMEM16A), including the cystic fibrosis transmembrane conductance regulator (CFTR) expressed on endo(sarco)plasmic reticulum. These findings demonstrate compartmentalization of Ca2+ signals dictates the odorant receptor OR2W3-induced ASM relaxation and identify a previously unrecognized E-C coupling mechanism that could be exploited in the development of therapeutics to treat obstructive lung diseases. [ABSTRACT FROM AUTHOR]

Species conservation efforts are increasingly using genetic diversity and phylogeny to inform policy decisions. Evolutionary distinctiveness (ED), which estimates genetic diversity based on phylogenetic branch lengths and species richness, is commonly used to prioritize species conservation. Large-scale studies of ED in amphibians have reported correlations among threatened status, area of distribution and habitat lost. However, conservation priorities based on ED could be more impactful for clades with high species richness, as well as considering the risk factors associated with species vulnerability in a local setting. We implemented comparative phylogenetic methods and multivariate regression to test the factors influencing species threatened status in a large clade of frogs within the subfamily Hylinae. We present the most comprehensive molecular phylogeny for the group, including 139 species, 265 individuals, and 12 additional species that were not previously studied. Additional data for each species includes (1) conservation status on the IUCN red list; (2) evolutionary distinctiveness; (3) species distribution in square kilometers; and (4) elevation profiles. Using a Phylogenetic General Linear Model to test the relationships among these variables, we found a significant correlation between threatened status and elevation. The threatened status of species was not correlated with ED or distribution area, which is inconsistent with large-scale studies spanning multiple families. By taking evolutionary history into account and testing for relationships between IUCN threatened status and factors at a regional scale, we provide new information for redirecting tree frog conservation efforts in the Neotropics. [ABSTRACT FROM AUTHOR]

Coronary artery disease is the leading cause of morbidity and mortality. Tools have been developed to accurately diagnose and evaluate coronary artery disease. Coronary CT angiography (CCTA) provides detailed imaging to deliver precise analysis and prognostic information. We sought to compare the radiation dose from a 256-detector-row CT scanner with that from a 64-detector-row CT scanner across a similar profile of patients. Methods: Consecutive patients were screened for the Converge Registry study and, after consenting to be included, were enrolled in accordance with an Institutional Review Board-approved protocol. A control group who underwent 64-row CCTA were matched by age, sex, and body mass index (BMI) with a group who underwent 256-row CCTA. Results: We compared 110 patients in each group. We found that mean dose-length product (DLP) was significantly lower in the 256-row group than in the 64-row group (P < 0.05). The radiation dose was reduced by 32% with use of the 256-row scanner for BMIs of 18.5-24.9 (DLP, 111.2 vs. 76.1 mGy-cm [1.56 vs. 1.07 mSv]; P < 0.05). For each BMI subgroup, there was a significant decrease in dose. Regression analysis found that with increasing BMIs, DLP significantly increased for both scanners. Conclusion: The 256-row scanner provided CCTA scans at significantly lower radiation doses than the 64-row scanner in different BMI groups, with all other variables accounted for. Lower radiation exposure along with lower contrast requirements can provide images with high diagnostic accuracy and less risk to the patient. [ABSTRACT FROM AUTHOR]

Background: Cubital tunnel syndrome is a common compressive neuropathy of the upper extremity. The anconeus epitrochlearis muscle is an unusual but occasional contributor. We review our experience with this anomalous muscle in elbows with cubital tunnel syndrome. Methods: We retrospectively reviewed charts of 13 patients noted to have an anconeus epitrochlearis muscle associated with cubital tunnel syndrome. Results: Ten patients had unilateral ulnar neuropathy supported by nerve conduction studies. Three had bilateral cubital tunnel syndrome symptoms with 1 of those having normal nerve conduction studies for both elbows. Eight elbows were treated with myotomy of the anconeus epitrochlearis muscle and submuscular transposition of the ulnar nerve. The other 8 elbows were treated with myotomy of the anconeus epitrochlearis muscle and in situ decompression of the ulnar nerve only. All but 1 patient had either clinical resolution or improvement of symptoms at follow-up ranging from 2 weeks to 1 year after surgery. The 1 patient who had persistent symptoms had received myotomy and in situ decompression of the ulnar nerve only. Conclusions: An anomalous anconeus epitrochlearis occasionally results in compression of the ulnar nerve but is usually an incidental finding. Its contribution to compression neuropathy can be tested intraoperatively by passively ranging the elbow while observing the change in vector and tension of its muscle fibers over the ulnar nerve. Regardless of findings, we recommend myotomy of the muscle and in situ decompression of the ulnar nerve. Submuscular transposition of the ulnar nerve may be necessary if there is subluxation. [ABSTRACT FROM AUTHOR]

Gap junction (GJ) channels are oligomers of connexins forming channels linking neighboring cells. GJs formed by different connexins show distinct unitary channel conductance (γj), transjunctional voltage-dependent gating (Vj-gating) properties, and modulation by intracellular magnesium ([Mg2+]i). The underlying molecular determinants are not fully clear. Previous experimental evidence indicates that residues in the amino terminal (NT) and initial segment of the first extracellular (E1) domain influence the γj, Vj-gating, and/or [Mg2+]i modulation in several GJs. Increasing negatively charged residues in Cx50 (connexin50) E1 (G46D or G46E) increased γj, while increasing positively charged residue (G46K) reduced the γj. Sequence alignment of Cx50 and Cx37 in the NT and E1 domains revealed that in Cx50 G8 and V53, positions are negatively charged residues in Cx37 (E8 and E53, respectively). To evaluate these residues together, we generated a triple variant in Cx50, G8E, G46E, and V53E simultaneously to study its γj, Vj-gating properties, and modulation by [Mg2+]i. Our data indicate that the triple variant and individual variants G8E, G46E, and V53E significantly increased Cx50 GJ γj without a significant change in the Vj gating. In addition, elevated [Mg2+]i reduced γj in Cx50 and all the variant GJs. These results and our homology structural models suggest that these NT/E1 residues are likely to be pore-lining and the variants increased the negative electrostatic potentials along the GJ pore to facilitate the γj of this cation-preferring GJ channel. Our results indicate that electrostatic properties of the Cx50 GJ pore are important for the γj and the [Mg2+]i modulation. [ABSTRACT FROM AUTHOR]

The synthesis and crystal structure of the title compound, C14H8Br2, is described. The molecule is positioned on a twofold rotation axis and the asymmetric unit consists of half a molecule with the other half being generated by symmetry. The presence of two large bromine atoms in the bay region significantly distorts the molecule from planarity and the mean planes of the two terminal rings of the phenanthrene system are twisted away from each other by 28.51 (14)°. The torsion angle between the two C--Br bonds is 74.70 (14)° and the distance between the two Br atoms is 3.2777 (13) Å. The molecules pack in layers in the crystal, with the centroids of the central rings of the phenanthrene units in adjacent layers separated by a distance of 4.0287 (10) Å. These centroids are shifted by 2.266 (6) Å relative to each other, indicating slippage in the stacking arrangement. Furthermore, the distance between the centroids of the terminal and central rings of the phenanthrene units in adjacent layers is slightly shorter at 3.7533 (19) Å. While all of the molecules within each layer are oriented in the same direction, those in adjacent layers are oriented in the opposite direction, leading to anti-parallel stacks. [ABSTRACT FROM AUTHOR]

Application of phosphate (P) fertilizers to farmland is projected to be causing worldwide accumulation of fluorine (F) in agricultural soils and associated ecosystems, but wide-scale field data have been lacking. We report results of sampling across two large regions of New Zealand involving soils of 298 native and farmed properties. This has enabled the first wide-scale retrospective estimates of anthropogenic F enrichment in pastoral and horticultural soils. Results validate earlier projections: F accumulation in farmed soils has been comparatively rapid (mean increase 2.1 % per year) and widespread. Over 50 years, average total F concentrations in surface soils have doubled from ~220 to 440 mg/kg. Thresholds protective against chronic fluorosis in grazing animals are being substantively passed, indicating that land may be rendered unsuitable for pastoral production in the mid-term future: when sampled 44 % of dairy farm soil samples exceeded 500 mg/kg total F, and 10 % exceeded 650 mg/kg. Potential for F toxicity to plants and wildlife appear to be significant and marginal, respectively, at current soil F concentrations. Relationships between F and other elements suggest P fertilizers are also a source of several elements geochemically linked to F, and that anthropogenic F modifies soil aluminium (Al) chemistry, potentially opening a broad-scale Al exposure pathway to pollinating insects. Findings signal a need for systematic examination of the range of possible affects that may be linked to increasing soil F, which include various types of ecotoxicity, altered Al chemistry, reduced P availability, and potential for increased dietary intakes of F, Al and aluminofluoride (AlF) species over time. [ABSTRACT FROM AUTHOR]

It has been previously shown that relatively simple computational fluid dynamics (CFD) models can be used to calculate the transfer impedances, including the associated end corrections, of microperforated panels. The impedance is estimated by first calculating the pressure difference across a single hole when a transient input velocity is imposed, and then Fourier transforming the result to obtain the impedance as a function of frequency. Since the size of the hole and the dimensions of the inlet and outlet channels are very small compared to a wavelength, the flow through the hole can be modeled as incompressible. By using those procedures, Bolton and Kim extended Maa's classical theory to include a resistive end correction for sharp-edged cylindrical holes which differs from those previously proposed by the inclusion of a static component. Here it is shown that CFD models can also be used to compute end corrections for tapered holes. Since practical experimental characterization of perforated materials often involves measurement of the static flow resistance, a closed form empirical equation for that quantity has been developed. Finally, it is shown that configurations having equivalent static flow resistances can yield different acoustic absorptions. [ABSTRACT FROM AUTHOR]

The "environmental dependency syndrome" refers to a loss of personal autonomy such that a person's environment almost entirely controls their actions. The goal of this study is to learn if patients with frontotemporal degeneration (FTD) exhibit prolonged TV watching, a behavior which may be a symptom of environmental dependency. We recruited 40 patients with FTD and 48 patients with Alzheimer's disease (AD), and asked these participants' caregivers about TV watching behaviors including total viewing time and channel/show preference, along with other behaviors indicative of environmental dependency. Compared to AD patients, FTD patients watched TV for a longer time. In addition, the patients who watched more TV showed more signs of environmental dependency. Increased TV watching may be a sign of environmental dependency, however further research is needed to explore other hypotheses. [ABSTRACT FROM AUTHOR]

The article discusses several rulings in 2012 by the U.S. Court of Appeals for the Federal Circuit in cases which deal with patent law, including information on legal proceedings which address topics such as patentability, patent infringement, and statutes of limitation. Other topics include remedies and damages in patent-related lawsuits, as well as the case Fox Group, Inc. v. Cree Inc. which deals with disclosure under Title 35, Section 102(g) of the U.S. Patent Act.

We compared the outcomes of intramedullary nailing with plate-screw fixation in the treatment for ipsilateral fracture of the hip and femoral shaft. A retrospective study. Level 1 Trauma. Forty-one patients (32 males and 9 females; mean age, 34 years; age range, 21-53) with ipsilateral hip and femoral shaft fractures were treated between 1995 and 2005. Eighteen patients were injured in motor vehicle accidents, and 23 fell from a height. All patients were treated by one of the two methods of internal fixation: a screw-plate fixation ( n = 24, Group I) or intramedullary nailing ( n = 17, Group II). The fracture union time, nonunion, delayed union, implant failure, need of further surgeries, and functional outcomes were investigated and compared. Fisher's exact test showed that Group I had a significantly higher frequency of nonunion than that of Group II ( P = 0.029). Although Group I had more nonunions, delayed unions, and revision operations than Group II, the total union time was similar for both groups. Intramedullary nailing was found to be superior to screw-plate fixation due to improved functional bearing, increased rate of union, stability, and mechanical solidity. The reconstruction nail method is an acceptable alternative treatment for ipsilateral hip and femoral shaft fractures. [ABSTRACT FROM AUTHOR]

Atherosclerosis is a major cardiovascular disease involving accumulations of lipids, white blood cells, and other materials on the inside of artery walls. Since the calcification found in the advanced stage of atherosclerosis dramatically enhances the mechanical properties of the plaque, restoring the original lumen of the artery remains a challenge. High-speed rotational atherectomy, when performed with an ablating grinder to remove the plaque, produces much better results in the treatment of calcified plaque compared to other methods. However, the high-speed rotation of the Rotablator commercial rotational atherectomy device produces microcavitation, which should be avoided because of the serious complications it can cause. This research involves the development of a high-speed rotational ablation tool that does not generate microcavitation. It relies on surface modification to achieve the required surface roughness. The surface roughness of the tool for differential cutting was designed based on lubrication theory, and the surface of the tool was modified using Nd:YAG laser beam engraving. Electron microscope images and profiles indicated that the engraved surface of the tool had approximately 1 μm of root mean square surface roughness. The ablation experiment was performed on hydroxyapatite/polylactide composite with an elastic modulus similar to that of calcified plaque. In addition, differential cutting was verified on silicone rubber with an elastic modulus similar to that of a normal artery. The tool performance and reliability were evaluated by measuring the ablation force exerted, the size of the debris generated during ablation, and through visual inspection of the silicone rubber surface. [ABSTRACT FROM AUTHOR]

The classical Maa theory for microperforated materials was initially formulated for constant diameter, cylindrical holes. Since then, a number of ad hoc corrections have been suggested to account for different hole shapes: in particular, rounding of the aperture. Here it is shown that the resistance and reactance of small apertures may be calculated using relatively simple CFD models in which a single hole is modelled. The fluid is assumed to be viscous but incompressible, and the geometry is assumed to be axisymmetric. It will be shown that this approach essentially reproduces the classical theory of Maa for circular, sharp-edged apertures. However, it will also be shown that the resistive end correction, in particular, exhibits a clear dependence on frequency and geometrical parameters that is neglected in conventional microperforated material models. [ABSTRACT FROM AUTHOR]

Abstract Laser-induced fluorescence (LIF) is an effective in-situ probe for NO concentrations below 300 ppm in a non-thermal plasma reactor. A new method has been developed to measure in-situ NO concentration in the reactor discharge region using a long-time—on the order of seconds—averaged fluorescence detection. This method, for quantifying NO concentration in a nonthermal plasma reactor, is simpler than a short-time—on the order of nanoseconds—fluorescence detection. For accurate measurement based on the new method, the LIF intensity must be close to the corona-induced fluorescence (CIF) intensity; the CIF intensity serves as a guide in selecting the LIF intensity. We find that a kinetic model proposed earlier works for two-tube reactors and represents the NO concentration in the middle of the reactor, which verifies the assumption of gas plug flow. [ABSTRACT FROM AUTHOR]

Background: Activation of free fatty acid receptors (FFAR1 and FFAR4) which are G protein-coupled receptors (GPCRs) with established (patho)physiological roles in a variety of obesity-related disorders, induce human airway smooth muscle (HASM) cell proliferation and shortening. We reported amplified agonist-induced cell shortening in HASM cells obtained from obese lung donors. We hypothesized that FFAR1 modulate excitation-contraction (EC) coupling in HASM cells and play a role in obesity-associated airway hyperresponsiveness.Methods: In HASM cells pre-treated (30 min) with FFAR1 agonists TAK875 and GW9508, we measured histamine-induced Ca2+ mobilization, myosin light chain (MLC) phosphorylation, and cortical tension development with magnetic twisting cytometry (MTC). Phosphorylation of MLC phosphatase and Akt also were determined in the presence of the FFAR1 agonists or vehicle. In addition, the effects of TAK875 on MLC phosphorylation were measured in HASM cells desensitized to β2AR agonists by overnight salmeterol treatment. The inhibitory effect of TAK875 on MLC phosphorylation was compared between HASM cells from age and sex-matched non-obese and obese human lung donors. The mean measurements were compared using One-Way ANOVA with Dunnett's test for multiple group comparisons or Student's t-test two-group comparison. For cortical tension measurements by magnetic twisted cytometry, mixed effect model using SAS V.9.2 was applied. Means were considered significant when p ≤ 0.05.Results: Unexpectedly, we found that TAK875, a synthetic FFAR1 agonist, attenuated histamine-induced MLC phosphorylation and cortical tension development in HASM cells. These physiological outcomes were unassociated with changes in histamine-evoked Ca2+ flux, protein kinase B (AKT) activation, or MLC phosphatase inhibition. Of note, TAK875-mediated inhibition of MLC phosphorylation was maintained in β2AR-desensitized HASM cells and across obese and non-obese donor-derived HASM cells.Conclusions: Taken together, our findings identified the FFAR1 agonist TAK875 as a novel bronchoprotective agent that warrants further investigation to treat difficult-to-control asthma and/or airway hyperreactivity in obesity. [ABSTRACT FROM AUTHOR]