Normal Contraction Research Articles

Explorations of Sodium‐Potassium Adenosine Triphosphatase, Na+K+ ATPase, and the Inhibitory Cardiac Glycoside, Ouabain

Sodium‐potassium adenosine triphosphatase (Na+K+ ATPase) resides in the plasma membranes of most animal cells, where it is involved in actively maintaining the cell's resting membrane potential. Though most commonly associated with its function in priming neurons for actions potentials, it is also essential for facilitating the transport of other molecules, regulating cell size, the transduction of particular intracellular signals, and for its role in cardiac muscle contraction. Ouabain, a plant derived cardiac glycoside has been found to interact with the cardiac specific isoform of Na+K+ ATPase with the indirect result of inducing stronger than normal muscle contractions. This property has led to ouabains use as a treatment for congestive heart failure. The Blue Valley North 2018–19 MSOE Center for BioMolecular Modeling SMART Team modeled the Na+K+ ATPase (PDB: 3KDP) using JMol with the goal of further exploring this protein's unique properties and its inhibition by ouabain and other toxins. Na+K+ ATPase is heterotrimeric complex formed from a, b, and g‐subunits. The a‐subunit contains the catalytic cytoplasmic and membrane domains. The catalytic domain is further subdivided into a nucleotide (N) binding domain responsible for binding ATP, the phosphorylation (P) domain that becomes phosphorylated upon ATP hydrolysis, and the actuator (A) domain responsible for translating conformational changes from the cytoplasmic portion of the protein to the membrane domain which controls the opening and occlusion of alternating half‐channels. Phosphorylation of the Na+K+ ATPase occurs at a highly conserved Asp369 residue found among P‐type pumps, while a conserved Glu found in the A domain is essential for dephosphorylation. Changes in the interactions between the three catalytic domains during phosphorylation and dephosphorylation are translocated into conformational changes in the membrane domain. The membrane domain is composed of 10 transmembrane helices (M1–M10), and contains the sites for ion binding and transport. The core membrane helices, M1–M6, contain two binding sites that are shared, asynchronously, by Na+ and K+. The third Na+ site is resides within the M7–M10 helices. Na+K+ ATPase cycles between two enzymatic states E1 and E2. The E1 state is characterized by a conformation with a half‐channel opened cytoplasmically, ATP and Na+ binding, and subsequent protein phosphorylation. In contrast, during the E2 state a half‐channel opens extracellularly, K+ binding occurs, and the protein is autodephosphoryled. Na+ export occurs during the E2 state while K+ import occurs during the E1 state, as these ions are exchanged with their counterpart. Overall, for each ATP hydrolyzed, 3 Na+ ions and 2 K+ ions are exchanged. Furthermore, the b‐subunit is facilitates appropriate membrane trafficking of the Na+K+ ATPase complex, while the g‐subunit is implicated in the regulation of transport.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Regulation of Lymphatic Muscle Cell Contractile Activity by Intracellular Calcium Signals

Lymphedema is characterized by chronic edema due to lymphatic insufficiency. Normally, the spontaneous contractions of the collecting lymphatic vessels (cLVs) function to pump lymph against a pressure gradient back to the blood stream, however cLVs from lymphedema patients often display weak or irregular pumping activity. We have recently shown that that mouse lymphatic muscle cells (LMCs) exhibit a diastolic depolarization that sets contraction frequency and is the basis of cLV pacemaking and autorhythmicity. In murine cLVs, this diastolic depolarization is pressure‐dependent and is mediated by the Ca2+ activated chloride channel Anoctamin1 (Ano1). Ano1 is presumed to be activated by the spontaneous Ca2+ transients originating from the sarcoplasmic reticulum (SR), however the identity of the SR Ca2+ channels underlying these diastolic Ca2+ transients has not been rigorously tested using genetic models. We tested the hypothesis that intracellular Ca2+ signals released from the SR are critical in regulating lymphatic pacemaking and contractile activity. We sorted murine LMCs and observed expression of both Itpr1 and RyR2. We tested the role of Itpr1 and RyR2 in the electrical and contractile regulation of the murine inguinal‐axillary cLV (Ing‐Ax) through the use of inducible deletion models (MYH11CreERT2) or mice harboring knock‐in gain of function (GOF) of loss of function (LOF) mutations. These mouse models include Ano1fl/fl, Itpr1fl/fl, Itpr1DK (GOF mutant), Itpr1DA (LOF mutant), RyR2fl/fl, and RyR2VF (GOF mutant). We performed ex vivo isobaric vessel myography in response to a physiological pressure range to determine Ing‐Ax contraction frequency (Freq), contraction amplitude (Amp), and vessel tone (Tone). In some experiments, we also recorded membrane potential using “sharp electrodes” and imaged Ca2+ signals using the genetically encoded cytosolic Ca2+ sensor GCaMP6f or GCaMP8 from pressurized Ing‐Ax cLVs. Ing‐Ax cLVs isolated from the heterozygous Itpr1DA/WT, the homozygous Itpr1DA/DA, and Itpr1smKO mice had significantly reduced contraction Freq and Tone, but also observed significantly higher Amp. Strikingly, Itpr1smKO Ing‐Ax cLVs also lacked the pressure‐dependent chronotropy similar to cLVs from Ano1smKO mice, however Itpr1smKO vessels had unstable periodicity in the contraction rhythm. Ing‐Ax cLVs from RyR2smKO mice had normal contraction Freq, Amp and Tone. In contrast, Ing‐Ax cLVs from the GOF RyR2VF/KO mice had significantly higher Tone and Freq, but significantly reduced Amp. Ing‐Ax cLVs from GOF Iptr1DK mice contracted into a paralytic rigor during the vessel warming and equilibration period likely due to Ca2+overload. These findings suggest that the pressure‐dependent Freq in murine cLVs is mediated by the activation of Ano1 by SR Ca2+ release through Itpr1 and an acceleration of the diastolic depolarization.Support or Funding InformationR01‐H122578 (MD), R01‐HL133256 & R01‐HL137745 (JJ)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Retinal vasodilatation in the affected eye but reduced pressure autoregulation of both eyes in unilateral Coats' disease.

Coats' disease is characterized by vascular hyperpermeability, oedema and accumulation of exudates related to impairment of retinal vascular function. The background for the development of the disease is unknown, but it is likely that the study of diameter changes of retinal vessels may contribute to understanding the pathophysiology of the disease. In seven patients with unilateral Coats' disease (mean age=34.7years, range: 11-69years), the baseline diameter and reactivity of retinal vessels during an increase in the arterial blood pressure by isometric exercise and in the metabolism by flicker stimulation were measured on video recordings of the retina obtained with the Dynamic Vessel Analyzer. The baseline diameter of retinal vessels was larger in the affected than in the unaffected eyes which was significant for the arterioles (p=0.02), but not for the venules (p=0.15). During an increase in the arterial blood pressure induced by isometric exercise, the normal contraction of arterioles was absent in both eyes (p>0.7), whereas there was a significant dilatation of the venules in the unaffected eyes (p=0.04). Stimulation with flickering light induced normal dilatation of retinal vessels in both affected and unaffected eyes. Unilateral Coats' disease is accompanied by vasodilatation in the affected eye but impaired pressure autoregulation in both eyes. A further investigation of the disease should include an elucidation of the background for dilatation of retinal vessels in affected eyes and whether impaired pressure autoregulation can be found in vessels elsewhere in the body.

C0 Ig-Domain of Cardiac Myosin Binding Protein-C Interacts with the Regulatory Light Chain of Myosin-S1 Bound to the Native Cardiac Thin Filament

Muscle contraction, relies on sliding of myosin-based thick filaments and thin filaments comprised of actin, tropomyosin and troponin. Cardiac myosin binding protein-C (cMyBP-C), an intrinsic protein of the thick filament, is a key regulator of actomyosin interactions, essential for both normal cardiac contraction and for increased cardiac contractility in response to inotropic stimuli. Mutations in MYBPC3, the gene encoding cMyBP-C, are the single most common genetic cause of hypertrophic cardiomyopathy (HCM). It has been proposed that the interaction of the N-terminal domains (NTDs) of cMyBP-C (e.g. C0, C1, M and C2) with myosin heads may stabilize the super-relaxed state (SRX-state) of the thick filament, while NMR experiments directly confirmed the binding of the C0 Ig-domain of cMyBP-C to the isolated regulatory light chain (RLC) of myosin. We recently demonstrated that C1 can activate the thin filament to the same extent as rigor myosin-S1, while C0 significantly enhances the activating effect of C1. Our data also show that C0 competes with myosin-S1 for binding to the thin filament. In order to understand how C0 interacts with the thick and thin filaments upon active cross-bridge formation we used cryo electron microscopy and image analysis of native cardiac thin filaments decorated with rigor myosin-S1 and the C0-domain of cMyBP-C. We show that C0 readily binds to the RLC of myosin-S1 bound to the thin filament in rigor state. Taking into account that the C0-domain is specific to the cardiac muscle and is absent in the skeletal MyBP-C we suggest a model of how the C0-domain of cMyBP-C participates in the regulation of cardiac contraction.

Creating a prototype of a robotic human hand for people with permanent disabilities

Robotics is the technology of designing, constructing and exploiting robots with major applications. One such application is to use robots in assisting people with physical disabilities. The goal of this work is to create a prototype of a robotic human hand. The basic components used in designing the hand prototype are Arduino UNO based on ATMEL microcontroller development board, breadboard for easy assembly of electronic circuits, jumper cables, 3D printed components from PLA and ABS, servo motors, reinforced cord, linking elements and other elements for stability of the prototype. For printing the 3D hand model, a 3D fused-deposition modelling printer with two plastic printing materials, PLA and ABS, was used. The software solution for controlling the fingers and the basic safety of overturning/under-turning the fingers is C-based. The program was compiled and uploaded using the built-in IDE for Arduino. The project was completed in a 2-months period time. The result is a working and anatomically correct prototype of the human hand. Presently, the prototype can simulate normal contraction and relaxation of the fingers digitus quartus manus and digitus minimus manus, also known as the ring and pinky finger. This initial development is used as a base for a large scale project for creating a modern and functional design of robotic human hand. Depending on the patient needs, an individual model will be designed, allowing customization, depending on the patient’s age, size and other factors, and will be produced. Acknowledgements This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 692097.

Hypoxic preconditioning ameliorates endometrial and myometrial damage and improves uterine function following prolonged hypoxia in nonpregnant rats.

Normal repeated uterine contractions are associated with uterine hypoxic stresses and uterine transplantation and severe bleeding during hysterectomy may lead to hypoxia and irreversible cellular damage. This study investigated the effects of short repeated hypoxic episodes on the structure and function of uterine tissues following sustained prolonged hypoxia. Small segments of uterine tissue were dissected from three groups of nonpregnant rats and mounted in a tissue bath system. Prolonged hypoxia markedly increased the infiltration of eosinophils into the myometrium and caused fibrotic stroma and degeneration of endometrial glands with marked infiltration of eosinophils into the endometrium compared to the control group. In addition, the mean myometrial contractile function significantly decreased to 69 ± 1% compared to 100% control with irregular and uncoordinated contractile activity (p < 0.01). Intriguingly, preconditioning with brief hypoxic episodes prevented the endometrial and myometrial degenerative changes. Although the mean myometrial contractile function decreased to 80 ± 3% during reoxygenation compared to the 100% control, the entire force was greater than the force in the non-preconditioned group (p < 0.01). These results provide compelling evidence that prolonged hypoxia exacerbates the degree of cellular damage and that preconditioning with repeated cycles of short hypoxia/reoxygenation can ameliorate cellular damage.

Arbitration agreement or service contract on dispute resolution

Arbitration agreement plays a vital role in arbitral proceedings, because the absence of arbitration agreement will lead to the invalidity of arbitral proceedings. Firstly, arbitration agreement figures out the name and type of the arbitration mechanism, then it clarifies parties’ requirements relating to the arbitration procedure including substantive law for the merit, procedural law for the arbitration proceedings, language of arbitration, number of arbitrators in the tribunal, locality of arbitration etc. In its essence, arbitration agreement not only describes the parties’ autonomy but also serves as a service contract (service contract on dispute resolution), accordingly arbitration organ will supply service on dispute resolution for parties. Unlike normal service contracts, autonomies of parties in service contract on dispute resolution, which indicates that arbitration organ is the service supplier, are established in two divergent stages. In the event of specific circumstances, although arbitration agreement has validity, the arbitration organ can refuse to become a service supplier.

The Predictive Factors of Abnormal Esophageal Motility in Systemic Sclerosis Patients by High Resolution Manometry

AbstractBackground: Esophageal motility changes occur in about 80% of systemic sclerosis (SSc) patients with decreasing the motility of the lower two-thirds of the esophagus and lower esophageal sphincter (LES) pressure.Aim of Study: Using high resolution manometry (HRM) to study the esophageal motility disorder in patients with systemic sclerosis to evaluate the predictive factors associated with esophageal affection.Patients and Methods: A prospective study was done on twenty female patients with SSc. Demographic data and esophageal symptoms including Gastroesophageal Reflux disease (GERD) and dysphagia were evaluated. All patients underwent HRM and High Resolution Computed Tomography (HRCT), Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), Hemoglobin level (Hgb), Forced Vital Capacity (FVC).Results: High resolution manometry showed normal upper esophageal contraction in 100% of patients. Middle and lower esophageal contraction were normal in 30%, hypoperestalsis in 5%, and aperestalsis in 65% of patients. Regarding the lower esophageal sphincter pressure, it showed normal pressure in 30%, hypotensive in 65% and hypertensive in 5% of patients. The esophageal involvement was GERD in 70% and dysphagia in 45% of patients. The pulmonary involvement was dyspnea in 70% and cough in 40% of patients. All patients had skin involvement. Abnormal motility was present in 14 out of 20 patients (70%) which was significantly related to Interstitial Lung Disease (SSc-ILD) (p= 0.001), SSc subtypes (p=0.03), dyspnea (p=0.01) and FVC (p=0.046). A significant relation between GERD and ILD (p<0.05) was found.Conclusion: This study confirmed an increased prevalence of esophageal motility disorders in SSc patients with and without esophageal symptoms.

Class II PI3Ks α and β Are Required for Rho-Dependent Uterine Smooth Muscle Contraction and Parturition in Mice.

Class II phosphoinositide 3-kinases (PI3Ks), PI3K-C2α and PI3K-C2β, are highly homologous and distinct from class I and class III PI3Ks in catalytic products and domain structures. In contrast to class I and class III PI3Ks, physiological roles of PI3K-C2α and PI3K-C2β are not fully understood. Because we previously demonstrated that PI3K-C2α is involved in vascular smooth muscle contraction, we studied the phenotypes of smooth muscle-specific knockout (KO) mice of PI3K-C2α and PI3K-C2β. The pup numbers born from single PI3K-C2α-KO and single PI3K-C2β-KO mothers were similar to those of control mothers, but those from double KO (DKO) mothers were smaller compared with control mice. However, the number of intrauterine fetuses in pregnant DKO mothers was similar to that in control mice. Both spontaneous and oxytocin-induced contraction of isolated uterine smooth muscle (USM) strips was diminished in DKO mice but not in either of the single KO mice, compared with control mice. Furthermore, contraction of USM of DKO mice was less sensitive to a Rho kinase inhibitor. Mechanistically, the extent of oxytocin-induced myosin light chain phosphorylation was greatly reduced in USM from DKO mice compared with control mice. The oxytocin-induced rise in the intracellular Ca2+ concentration in USM was similar in DKO and control mice. However, Rho activation in the intracellular compartment was substantially attenuated in DKO mice compared with control mice, as evaluated by fluorescence resonance energy transfer imaging technique. These data indicate that both PI3K-C2α and PI3K-C2β are required for normal USM contraction and parturition mainly through their involvement in Rho activation.

Clinical Presentation of Hypoparathyroidism.

Parathyroid hormone (PTH) is one of the major hormones that regulates serum calcium. Hypoparathyroidism occurs when PTH secretion is insufficient. The main symptoms of hypoparathyroidism are the result of low blood calcium levels, hypocalcemia, which interferes with normal muscle contraction and nerve conduction. As a result, people with hypoparathyroidism can experience paresthesia, an unpleasant tingling sensation around the mouth and in the hands and feet, as well as muscle cramps and severe spasms known as "tetany" that affect the hands and feet. Many also report a number of other subjective symptoms. Hypocalcemia can be the cause of medical emergencies, for example seizures, severe irregularities in the normal heart beat, as well as laryngospasm, stridor, bronchospasm, and wheezing.

Comparison of Dinoprostone Vaginal Tablet and Vaginal Insert in Primigravid Women for Induction of Labor

Background: Labor of induction by use of different drugs is generally preferred for avoiding complications with prolonged pregnancy. Dinoprostone vaginal gel or insert tablets are commonly used for inducing contractions similar to normal delivery contractions. The current study compares the effectiveness of dinoprostone vaginal tablet and dinoprostone vaginal insert in induction of labor for primigravid women. Methods: The participants of the prospective cohort observational study were primigravid women. BISHOP score was used as a tool for predicting patient who required labor induction. All the participants with BISHOP score less than 6 were given either dinoprostone vaginal tablet or dinoprostone vaginal insert for induction of labor. Chi-square statistical analysis was performed using SPSS, version 17.0. Results: A total of 135 patients were studied. Post-term pregnancy was found to be most common indication among studied patients. Labor induction was executed by using dinoprostone vaginal tablet and insert in 61% and 31% patients, respectively. Statistically, no significant differences were found in the rate of cesarean section among two treatment regimens on applying Chi-square analysis. On average, two dinoprostone tablets per patient as compared to one vaginal insert were used for labor induction. Conclusion: Dinoprostone vaginal tablet and vaginal insert both have similar efficacy in induction of labor to be used in primigravid women. J Clin Gynecol Obstet. 2018;7(2):52-56 doi: https://doi.org/10.14740/jcgo471w

Transient uterine contractions as a potential pathology mimic on premenopausal pelvic MRI and the role of routine repeat T2 sagittal images to improve observer confidence.

Pelvic MRI has an increasingly important role in the evaluation of non-malignant uterine pathology including uterine leiomyomas, adenomyosis and endometriosis. Normal physiological myometrial junctional zone transient contractions can also be identified on MRI and have the potential to act as pathology mimics. This study aims to evaluate both the incidence of visible physiological contractions in premenopausal female pelvic MRI and also to support the routine acquisition of a repeat T2 sagittal sequence to differentiate transient physiological contractions from true underlying pathology and therefore improve observer confidence. A total of 279 female patients of child-bearing age who had undergone a pelvic MRI over a 16month period met the inclusion criteria. All patients underwent a standard examination protocol on the same hardware. This included performing two separate T2-weighted sagittal sequences as part of the protocol firstly as the initial and then as the final series for the examination. The sagittal series were reviewed separately by four readers and conclusions made for each case with regards to the presence of identifiable contractions on one or both series and their potential to act as pathology mimics. Of the 279 cases, there were 34 cases (12.2%) that were found to have transient junctional zone contractions acting as pathology mimics, resembling either leiomyomata or adenomyosis. Standard MRI sequences need to be able to distinguish normal transient physiological uterine contractions from true pathology to avoid diagnostic error. The routine utility of a repeat T2-weighted sagittal sequence performed at the conclusion of a patient's examination was shown to improve reader confidence in distinguishing transient contractions from true uterine pathology while adding minimal time penalty to the overall examination. It is therefore advocated that all premenopausal female pelvic MRI cases have a T2 sagittal series as the initial and then the final series as part of a routine protocol.

High-pressure synchrotron X-ray diffraction study of tremolite and actinolite in various fluids

Pressure-dependent structural and morphological changes of two amphibole minerals, tremolite and actinolite, were investigated up to 7.0 GPa using synchrotron X-ray powder diffraction underthree different pressure transmission media (PTM): water (W), CO2 and silicone oil (SI). The elastic response of tremolite and actinolite are found to be dependent on the PTM used. When using water (W) as PTM, tremolite and actinolite show normal volume contractions with bulk moduli of 74(1) and 78(1) GPa, respectively. When using CO2 as PTM, we observe the formation of calcite from tremolite above 3.8(1) GPa, whereas actinolite did not show any carbonation reaction. Under silicone oil PTM, we observe modulated volume contraction behaviors in both samples, compared to water and CO2 PTM, with bulk moduli in the order of 90(1) and 94(4) GPa for tremolite and actinolite, respectively.

Normal Stresses, Contraction, and Stiffening in Sheared Elastic Networks.

When elastic solids are sheared, a nonlinear effect named after Poynting gives rise to normal stresses or changes in volume. We provide a novel relation between the Poynting effect and the microscopic Grüneisen parameter, which quantifies how stretching shifts vibrational modes. By applying this relation to random spring networks, a minimal model for, e.g., biopolymer gels and solid foams, we find that networks contract or develop tension because they vibrate faster when stretched. The amplitude of the Poynting effect is sensitive to the network's linear elastic moduli, which can be tuned via its preparation protocol and connectivity. Finally, we show that the Poynting effect can be used to predict the finite strain scale where the material stiffens under shear.

On design of a three- class ECG classifier

Objectives: Manual analysis of ECG, specifically the ambulatory type, is tedious and time consuming, hence automation is desired. In this paper the authors have focused on the design of a three class ECG classifier using feature extraction and artificial neural networks. Once feature extraction is done, ANNs can be trained to classify the patterns reasonably accurately. Arrhythmia is one such type of abnormality detect able by an ECG signal. The three classes of ECG signals are Normal, Fusion and Premature Ventricular Contraction (PVC). The task of an ANN based system is to correctly identify the three classes, most importantly the PVC type, this being a fatal cardiac condition. Methods: The ECG data is taken from MIT-BIH Arrhythmia database. Fifty-five different feature extraction schemes are examined, along with a compact set of statistical morphological features and a reasonably accurate and fast classifier is designed. These feature extraction techniques coupled with ten morphological features have not been collectively studied and compared in literature so far. Findings: Multilayer perceptron with momentum learning rule is found to be the best classifier topology, while the best performing feature extraction schemes are: bior2.2, coif1, db9, rbior1.1, sym2, DCT and PCA. Application/Improvements: The reported findings can be effectively used for automated ECG arrhythmia classification for rapid analysis by a cardiac specialist thus saving time and arriving at a quick and reliable diagnosis. A similar approach can be used for designing an ECG classifier for more number of arrhythmia conditions. Keywords: Arrhythmia, Artificial Neural Networks, ECG Classifier Design, Wavelets

Adipose-derived stem cells improve full-thickness skin grafts in a rat model

To investigate the effects of heterologous adipose-derived stem cells (ADSCs) on autologous full-thickness skin grafts, we designed a first-intention healing model using Wistar rats. We harvested and sutured two full-thickness skin grafts in the dorsal recipient beds of 15 rats, randomized into three groups. In the treatment group, 1 × 106 ADSCs resuspended in saline solution (200 μL) were administered subcutaneously to the skin graft. The control group received only saline solution subcutaneously, whereas the negative control group did not receive any treatment. Compressive dressings were maintained until postoperative day 5. The grafts were assessed by two observers, who checked for the presence of epidermolysis on day 14. Planimetry showed the relative areas of normal skin, redness, ulceration, and contraction. Graft samples were obtained on day 14 and stained with hematoxylin and eosin and Masson's trichrome. Epidermal analysis evaluated thickening, keratosis, acanthosis, hydropic degeneration, and inflammatory infiltrate. Dermal evaluation investigated the absence of hair follicles, granulation tissue formation, presence of inflammatory infiltrate, and collagen deposition. Immunohistochemistry was performed for dermal anti-VEGF and epidermal anti-Ki-67 staining. The ADSC group presented better macroscopic aspects, lower incidence of epidermolysis, and less loss of hair follicles. In addition, the ADSC group presented the lowest frequency of histopathological changes in the dermis and epidermis, as well as the largest subcutaneous and granulation tissue VEGF averages and the weakest Ki-67 staining of the epidermal basal layer. Subcutaneous administration of ADSCs may improve the integration of skin grafts, reducing the deleterious effects of ischemia and reperfusion injury.

MUSCLEMOTION: A Versatile Open Software Tool to Quantify Cardiomyocyte and Cardiac Muscle Contraction In Vitro and In Vivo.

There are several methods to measure cardiomyocyte and muscle contraction, but these require customized hardware, expensive apparatus, and advanced informatics or can only be used in single experimental models. Consequently, data and techniques have been difficult to reproduce across models and laboratories, analysis is time consuming, and only specialist researchers can quantify data. Here, we describe and validate an automated, open-source software tool (MUSCLEMOTION) adaptable for use with standard laboratory and clinical imaging equipment that enables quantitative analysis of normal cardiac contraction, disease phenotypes, and pharmacological responses. MUSCLEMOTION allowed rapid and easy measurement of movement from high-speed movies in (1) 1-dimensional in vitro models, such as isolated adult and human pluripotent stem cell-derived cardiomyocytes; (2) 2-dimensional in vitro models, such as beating cardiomyocyte monolayers or small clusters of human pluripotent stem cell-derived cardiomyocytes; (3) 3-dimensional multicellular in vitro or in vivo contractile tissues, such as cardiac "organoids," engineered heart tissues, and zebrafish and human hearts. MUSCLEMOTION was effective under different recording conditions (bright-field microscopy with simultaneous patch-clamp recording, phase contrast microscopy, and traction force microscopy). Outcomes were virtually identical to the current gold standards for contraction measurement, such as optical flow, post deflection, edge-detection systems, or manual analyses. Finally, we used the algorithm to quantify contraction in in vitro and in vivo arrhythmia models and to measure pharmacological responses. Using a single open-source method for processing video recordings, we obtained reliable pharmacological data and measures of cardiac disease phenotype in experimental cell, animal, and human models.

Site Specific Evaluation of Lymphatic Vessel Sclerosis in Lower Limb Lymphedema Patients.

Histological changes in the collecting lymphatics in patients with lymphedema are classified as Normal type, Ectasis type, Contraction type, and Sclerosis type (NECST) classification. In this study, we investigated the condition of the lymphatic vessels in different sites of the legs. We prospectively investigated the lymphatic vessels of patients with lymphedema who underwent lymphaticovenous anastomosis (LVA) from August 8, 2014 to August 4, 2015 based on the NECST classification. Lymphedema was diagnosed using lymphoscintigraphy, indocyanine green (ICG) lymphography, and the International Society of Lymphology (ISL) Classification. The affected limbs were divided into four sites: proximal thigh (Site 1), distal thigh (Site 2), proximal crus (Site 3), and distal crus (Site 4). A total of 109 patients (205 limbs and 1028 lymphatics) were included in this study. Of the 109 patients, there were 100 women and 9 men with an average age of 61 years. The ratio of Ectasis type vessels increased toward the distal end of the limb with the highest occurrence rate being 54% at Site 4. As ISL stage, ICG stage, and lymphoscintigraphy stage advanced, so too did the ratio of Sclerosis type. In secondary lymphedema patients with lymphedema, the ratio of Ectasis type was more predominant in the distal end of the limb, whereas this tendency was not observed in primary lymphedema patients. Sclerotic lymphatics are more predominantly found in the proximal limb whereas nonsclerotic vessels are more often found toward the distal end. These findings help lymphatic surgeon determine incision sites.

Effects of Superficial and Deep Dry Needling on Pain and Muscle Thickness in Subject with Upper Trapezius Muscle Myofascial Pain Syndrome

Background: Dry needling is one of the main therapeutic approaches in patients with Myofascial pain syndrome. Few studies have been compared the superficial and deep dry needling methods in these patients. Objective: To evaluate the effects of superficial and deep dry needling on pain and muscle thickness in subjects with upper trapezius myofascial pain syndrome. Design: A randomized quasi-experimental double-blinded trial. Methods: 50 subjects with upper trapezius myofascial pain syndrome (age=26/08 ± 4/62, weight=63/88 ± 8/71 kg, height=167/7 ± 4/82 cm, pain duration=9/75 ± 7/05 m) randomly assigned to the superficial (n=25) and deep (n=25) dry needling groups. The pain and maximum thickness of upper trapezius muscle in rest, fair and normal contractions were measured by visual analogue scale (VAS) and an ultrasound device respectively before and after the intervention as well as 7 and 15 days follow-up. Results: The mixed-model ANOVAs revealed a significant group-by-time interaction (F=44.03, p<0.001) for pain and muscle thickness in rest (F=67.00, p<0.001), fair (F=108.73, p<0.001) and normal contraction (F=17.73, p<0.001). The main effects of group and time were statistically significant for pain, rest, fair and normal muscle thickness (p<0.001). There were not any significant differences in rest, fair and normal muscle thickness after intervention as well as 7 and 15 days follow-up. Conclusion: Both superficial and deep dry needling techniques induced significant short-term changes in the VAS. Muscle thickness in rest, fair and normal contractions did not show any significant changes between the groups.

361: Risk of cesarean delivery during the active phase of labor and uterine contractile activity

The diagnosis of labor arrest relies on a determination of normal labor progress and contraction adequacy. Using a modern framework of labor progress, we sought to identify contraction patterns that were associated with cesarean (CD) in the active phase of labor (>6 cm). Within a prospective cohort of nulliparous women at term who had an intrauterine pressure catheter in place, we included women who reached >6 cm dilation. Excluded were women with multifetal gestations. Women who had an active phase CD were compared to women who reached complete dilation. Measures of uterine activity were compared between the groups, including frequency (contractions/10 min), Montevideo units (MVU), Alexandria units (AU) (frequency x amplitude x duration), contraction shape (F:R ratio) (time from contraction peak to baseline return/time from start of contraction to peak), F:R ratio x amplitude, relaxation time, and resting pressure-min (relaxation time x baseline uterine tone). Logistic regression was used to adjust for confounding factors. Receiver operating characteristic curves were used to compare the different measures of uterine activity during the active phase of labor. Forty-three women had an active phase CD while 325 women reached complete dilation. Women with an active phase CD were more likely to be obese, black race, have an infant weighing ≥4000 gms, and preeclampsia or gestational hypertension. Contraction frequency, MVUs and AUs were significantly lower among women who had an active phase CD compared to women who reached complete dilation. Among women who achieved complete dilation, only 39.9% had MVUs >200 during the active phase. Contraction frequency and MVUs were similar in their use as predictive measures of active phase CD. Once nulliparous women reach the active phase of labor, contraction frequency and MVUs are most closely associated with risk of CD. Yet, a minority of women who achieve complete dilation have MVUs >200, which raises the question of whether this measurement should be a principal factor in the diagnosis of labor arrest.View Large Image Figure ViewerDownload Hi-res image Download (PPT)