Smooth muscle contraction regulates the size of the blood vessel lumen which directly affects the mechanical response of the vessel. Folding in arteries has been observed in arteries during excessive contraction, known as a coronary artery spasm. The interplay of muscle contraction, geometry, and material responses and their effects on stability can be understood through mathematical models. Here, we consider a three-layer cross-sectional model of a coronary artery with anisotropic properties and intimal thickening, and perform a linear stability analysis to investigate the circumferential folding patterns that emerge due to muscle contraction. Our model shows that a critical level of contractile activity yields a uniform strain distribution across the arterial wall. When the muscle is contracted above this critical level, the tissue behaves isotropically and it is more prone to circumferential instability. This theoretical framework could serve as a valuable tool to understand the relationship between arterial lumen morphology and wall contraction in health and disease.

Thesis (doctoral)--Lund University, 1998. / Added t.p. with thesis statement inserted. Includes bibliographical references.

Thesis (doctoral)--Lund University, 1998. / Added t.p. with thesis statement inserted. Includes bibliographical references.

Regulation of smooth muscle contraction depends on the phosphorylated state of myosin light chain (MLC). Although there are many kinases responsible for phosphorylating MLC, the myosin phosphatase complex is solely accountable for its dephosphorylation. Myosin phosphatase, in turn, is tightly regulated by many proteins. One of them being the CPI-17 gene family, which inhibits myosin phosphatase. This family of proteins is composed of CPI-17 itself, PHI-1, KEPI, and GBPI. Zebrafish have two genes each of CPI-17 and PHI-1, which are expressed during early embryonic development. This study sets out to investigate whether the two isoforms of CPI-17 and PHI-1 have diverged in function or expression using zebrafish as a model organism. Through a series of biochemical tests and assays, we have determined that the two isoforms have diverged in their expression pattern from each other, however they have similar function.

Indiana University-Purdue University Indianapolis (IUPUI) / The contractile activation of airway smooth muscle tissues stimulates actin polymerization and the inhibition of actin polymerization inhibits tension development. Actin depolymerizing factor (ADF) and cofilin are members of a family of actin–binding proteins that mediate the severing of F–actin when activated by dephosphorylation at serine 3. The role of ADF/cofilin activation in the regulation of actin dynamics and tension development during the contractile activation of airway smooth was evaluated in intact canine tracheal smooth muscle tissues. Two–dimensional gel electrophoresis revealed that ADF and cofilin exist in similar proportions in the muscle tissues and that approximately 40% of the total ADF/cofilin in unstimulated tissues is phosphorylated (inactivated). Phospho–ADF/cofilin decreased concurrently with tension development in response to stimulation with acetylcholine (ACh) or potassium depolarization indicating the activation of ADF/cofilin. Expression of an inactive phospho–cofilin mimetic (cofilin S3E), but not WT cofilin in the smooth muscle tissues inhibited endogenous ADF/cofilin dephosphorylation and ACh–induced actin polymerization. Expression of cofilin S3E in the tissues depressed tension development in response to ACh, but it did not affect myosin light chain phosphorylation. The ACh–induced dephosphorylation of ADF/cofilin required the Ca2+–dependent activation of calcineurin (PP2B). Expression of Slingshot (SSH) inactive phosphatase (C393S) decreased force development and cofilin dephosphorylation. Activation of ADF/cofilin was also required for the relaxation of tracheal muscle tissues induced by forskolin and isoproterenol. Cofilin activation in response to forskolin was not Ca2+–dependent and was not inhibited by calcineurin inhibitors, suggesting it was regulated by a different mechanism. Cofilin activation is required for actin dynamics and tension development in response to the contractile stimulation of tracheal smooth muscle and is regulated by both contractile and relaxing stimuli. These concepts are critical to understanding the mechanisms of smooth muscle contraction and relaxation, which may provide novel targets for therapeutic intervention in the treatment of abnormal airway responsiveness.

Indiana University-Purdue University Indianapolis (IUPUI) / Asthma, a chronic inflammatory disease of the airways, affects nearly 25 million Americans. The vast majority of these patients suffer from exercise-induced bronchoconstriction (EIB), a complication of asthma. Although traditionally treated pharmacologically, nutritional strategies provide a promising alternative for managing EIB as the prevalence of asthma may be due in part to changes in diet. Our objective was to determine the effects of novel nutritional strategies on hyperpnea-induced bronchoconstriction (HIB) in asthmatic individuals. HIB uses rapid breathing to identify EIB in a research or clinical setting. Fish oil, a combination of the omega-3 fatty acids eicosapentaenoic acid (EPA) and docsahexaenoic acid (DHA), has been shown to be effective in suppressing EIB. However, its use in combination with other nutritional supplements, the optimal fish oil formula, and its effect on smooth muscle contractility have not been fully explored. An in vivo study (study 1) was conducted in individuals with both asthma and HIB to determine whether a combination of fish oil and vitamin C was more effective than either one alone in alleviating HIB. Pulmonary function was significantly improved with both fish oil and the combination treatment but not with vitamin C alone. In study 2, individuals with both asthma and HIB were supplemented with DHA alone since the optimal formula for fish oil has yet to be ascertained; previous in vitro studies have suggested DHA may be the more potent omega-3 fatty acid in fish oil. However, no significant changes in pulmonary function or airway inflammation were seen with DHA supplementation. For study 3, canine airway smooth muscle tissue was treated with fish oil to determine the in vitro effect of fish oil on smooth muscle contractility. Acute treatment with fish oil relaxed smooth muscle strips that had been contracted with 5-hydroxytryptamine. These minor relaxations in smooth muscle tension with fish oil may represent significant changes at the level of the smaller airways. These studies have confirmed that fish oil represents a viable treatment modality for asthmatic individuals with EIB and suggest that fish oil may influence airway smooth muscle contractility.

The purpose of this manuscript is to establish a unified theory of porohyperelasticity with transport and growth and to demonstrate the capability of this theory using a finite element model developed in MATLAB. We combine the theories of volumetric growth and mixed porohyperelasticity with transport and swelling (MPHETS) to derive a new method that models growth of biological soft tissues. The conservation equations and constitutive equations are developed for both solid-only growth and solid/fluid growth. An axisymmetric finite element framework is introduced for the new theory of growing MPHETS (GMPHETS). To illustrate the capabilities of this model, several example finite element test problems are considered using model geometry and material parameters based on experimental data from a porcine coronary artery. Multiple growth laws are considered, including time-driven, concentrationdriven, and stress-driven growth. Time-driven growth is compared against an exact analytical solution to validate the model. For concentration-dependent growth, changing the diffusivity (representing a change in drug) fundamentally changes growth behavior. We further demonstrate that for stress-dependent, solid-only growth of an artery, growth of an MPHETS model results in a more uniform hoop stress than growth in a hyperelastic model for the same amount of growth time using the same growth law. This may have implications in the context of developing residual stresses in soft tissues under intraluminal pressure. To our knowledge, this manuscript provides the first full description of an MPHETS model with growth. The developed computational framework can be used in concert with novel in-vitro and in-vivo experimental approaches to identify the governing growth laws for various soft tissues.

The chicken ductus arteriosi (DA) are two embryonic blood vessels that shunt blood away from the non-ventilated lungs and towards the body and chorioallantoic membrane. I show that prostaglandins have a diminished role in maintaining chicken DA patency and nitric oxide inhibits oxygen induced contraction of the day 19 proximal DA in a time dependent manner. The pathways governing oxygen induced contraction in the chicken DA are similar to those found in mammals and include contributions from ROS, Kv channels, L-type Ca2+ channels, and the Rho kinase pathway. Longer exposure to high oxygen generates increased oxygen induced constriction of the day 19 DA that may be mediated through the Rho kinase pathway.

Increased airway smooth muscle (ASM) mass contributes to airway hyperresponsiveness (AHR) in asthma and is orchestrated by growth factors, cytokines and chemokines. Airway contractile responses are influenced by neuromediators, such as acetylcholine, and glutamate released by parasympathetic and sympathetic airway nerves. Hyperactivity of these neural elements, termed neurogenic inflammation, is linked with hypercontractility and AHR. Glutamate is a non-essential amino acid derivative, and its physiological role is traditionally considered with respect to its being the primary excitatory neurotransmitter in brain, and regulation of neuronal development and memory. In allergic inflammation, immune cells including dendritic cells, neutrophils and eosinophils, constitutively synthesize and release glutamate, which signals through activation of glutamate receptors, most important among which are ionotrophic N-methyl D-aspartate receptors (NMDA-R). We hypothesized that glutamatergic signaling mediated through NMDA-Rs plays an important role in inducing functional Ca2+ responses in human (H) ASM cells that can underpin airway hypercontractility. We investigated the expression and function of NMDA-Rs in HASM cells, and assessed the effects of pro-inflammatory cytokines on NMDA-R expression and functional responses. Moreover, we measured airway responses to NMDA in mice, murine thin cut lung slice preparations, and floating collagen gels seeded with HASMs. Our data reveal that airway myocytes express multi-subunit NMDA-R complexes that function as receptor-operated calcium channels (ROCCs), mobilizing intracellular Ca2+ in ASM in vitro and airway contraction ex vivo. Individual airway myocytes treated with NMDA-R agonist exhibit disparate temporal patterns of intercellular Ca2+ flux that can be partitioned into four discrete function sub-groups. Further we show that tumor necrosis factor (TNF) exposure modulates NMDA-R subunit expression, and these changes are associated with a shift in the distribution of myocytes in individual Ca2+-mobilization sub-groups in vitro. Further, post-TNF exposure, NMDA-R agonists’ treatment induced Ca2+-dependent airway dilation in murine lung slice preparations, an effect that was prevented by co-treatment with inhibitors of nitric oxide synthase (NOS) or cyclooxygenase (COX). Taken together, we conclude that NMDA-R regulate HASM-mediated airway contraction and their role can be affected upon exposure to asthma-associated inflammatory mediators. Thus, NMDA-Rs are of relevance to mechanisms that determine airway narrowing and AHR associated with chronic respiratory diseases. / October 2015

Šio eksperimentinio darbo tikslas buvo įvertinti 1,4-dihidropiridino ir benzimidazolo junginių poveikį jūros kiaulyčių širdies papiliarinių raumenų izometrinei funkcijai bei žmogaus kraujagyslių segmentų susitraukimui ir atsipalaidavimui. Buvo atlikti eksperimentiniai tyrimai su izoliuotais jūros kiaulyčių širdies papiliarinių raumenų preparatais ir izoliuotais žmogaus v.saphena magna ir a.thoracica interna kraujagyslių segmentais. Kraujagyslių preparatai gauti iš pacientų, kuriems buvo atliekamos širdies vainikinių arterijų jungčių suformavimo operacijos LSMU ligoninės Kauno klinikos Kardiochirurgijos klinikoje. Žmogaus izoliuotų kraujagyslių preparatų tyrimams išduotas Kauno regioninio bioetikos komiteto leidimas Nr. BE–2–64, data 2010–11–05. Buvo tiriamas 1,4-dihidropiridinų bei benzimidazolo junginių poveikis izometrinei jūros kiaulyčių širdies papiliarinių raumenų funkcijai, registruojant preparatų elektromechaninį susitraukimo jėgos ir transmembraninio veikimo potencialus. Eksperimentiniai tyrimai, atlikti in vitro sąlygomis, įvertinant 1,4-dihidropiridinų bei benzimidazolo junginių poveikį žmogaus izoliuotų kraujagyslių (v.saphena magna ir a.thoracica interna) segmentų susitraukimui ir atsipalaidavimui, skirtingų ekstraląstelinio Ca2+ koncentracijų įtaką tiriamųjų junginių poveikiui kraujagyslių segmentų susitraukimui ir atsipalaidavimui. Tiriamieji 1,4–dihidropiridinų junginiai sintezuoti Latvijos Organinės Sintezės institute. / This study aim was to evaluate the effects of 1,4-dihydropiridines and benzimidazole deriva¬tives on the isometric function of guinea pig papillary cardiac muscles and the contraction and relaxation of vascular segments in humans. The experiments were carried out on isolated samples of human great saphenous vein (v. saphena magna) and internal thoracic artery (a. tho¬racica interna). The vein and arteries samples were taken from patients who underwent coronary artery bypass. The study was approved by the Regional Ethics Committee of the Biomedical Research on 05/11/2010, license No. BE-2-64, Kaunas, Lithuania. The inotropic activity and transmembrane AP duration of the dihydropyridine derivatives were evaluated on the guinea-pig papillary muscles and aorta vascular samples. Synthesis of 1,4-dihydropyridine derivatives were performed in Latvian Organic Synthesis institute. During this study were performed the evaluation of 1,4-dihydropyridines and benzimidazole derivatives on contraction force and action potential in guinea pig papillary muscles. Effects of 1,4-dihydropyridines and benzimidazole derivatives on contraction and relaxation of the segments of the great sa¬phenous vein (v. saphena magna) and internal thoracic artery (a. tho¬racica interna) in humans, the extra-cellular concentration of Ca2+ and the effect of the studied compounds and were evaluated. Also assessment of preventive effect of the calcium channel blockers and benzimidazole derivative on contraction... [to full text]