Trendelenburg gait after total hip arthroplasty due to reduced muscle contraction of the hip abductors and extensors.

Background: Despite experiencing pain relief and improved activities of daily living after total hip arthroplasty (THA) for osteoarthritis of the hip, a Trendelenburg gait may be observed in some patients. The concentric and eccentric contraction patterns of hip muscles in a Trendelenburg gait are not well understood. Methods: This study included 89 patients (28 males and 61 females, mean age 66.5 ± 8.4 years, mean postoperative period 1.3 ± 0.4 years) after unilateral THA without functional impairment on the contralateral side. Gait analysis utilized a three-dimensional motion capture system to assess pelvis and hip angles, hip moment, and hip power. A Trendelenburg gait was defined as positive when nonoperative pelvic descent occurred at 30 % of the gait cycle, equivalent to mid-stance. Patients were classified into Trendelenburg gait-positive and -negative groups for statistical analysis. Unpaired t-test and chi-square test were used to compare the two groups. Multiple regression analysis was conducted to identify factors associated with the presence of a Trendelenburg gait. Results: A Trendelenburg gait was observed in 24 patients (27 %). Multiple regression analysis indicated that abduction (p < 0.01) and extension (p = 0.03) of hip joint power were significant determining of a Trendelenburg gait. Patients with a Trendelenburg gait exhibited reduced eccentric contraction of the hip abductor muscles and decreased concentric contraction of hip extensor muscles during early to mid-stance of their gait. Conclusion: Centrifugal contraction of hip abductor muscles and diminished eccentric contractility of hip extensor muscles appear crucial for hip stabilization mechanisms during gait after THA.

Mechanisms of myosin II force generation: insights from novel experimental techniques and approaches.

Myosin II is a molecular motor that converts chemical energy derived from ATP hydrolysis into mechanical work. Myosin II isoforms are responsible for muscle contraction and a range of cell functions relying on the development of force and motion. When the motor attaches to actin, ATP is hydrolyzed and inorganic phosphate (Pi) and ADP are released from its active site. These reactions are coordinated with changes in the structure of myosin, promoting the so-called "power stroke" that causes the sliding of actin filaments. The general features of the myosin-actin interactions are well accepted, but there are critical issues that remain poorly understood, mostly due to technological limitations. In recent years, there has been a significant advance in structural, biochemical, and mechanical methods that have advanced the field considerably. New modeling approaches have also allowed researchers to understand actomyosin interactions at different levels of analysis. This paper reviews recent studies looking into the interaction between myosin II and actin filaments, which leads to power stroke and force generation. It reviews studies conducted with single myosin molecules, myosins working in filaments, muscle sarcomeres, myofibrils, and fibers. It also reviews the mathematical models that have been used to understand the mechanics of myosin II in approaches focusing on single molecules to ensembles. Finally, it includes brief sections on translational aspects, how changes in the myosin motor by mutations and/or posttranslational modifications may cause detrimental effects in diseases and aging, among other conditions, and how myosin II has become an emerging drug target.

A multimodal deep learning-based algorithm for specific fetal heart rate events detection.

OBJECTIVES: This study aims to develop a multimodal deep learning-based algorithm for detecting specific fetal heart rate (FHR) events, to enhance automatic monitoring and intelligent assessment of fetal well-being. METHODS: We analyzed FHR and uterine contraction signals by combining various feature extraction techniques, including morphological features, heart rate variability features, and nonlinear domain features, with deep learning algorithms. This approach enabled us to classify four specific FHR events (bradycardia, tachycardia, acceleration, and deceleration) as well as four distinct deceleration patterns (early, late, variable, and prolonged deceleration). We proposed a multi-model deep neural network and a pre-fusion deep learning model to accurately classify the multimodal parameters derived from Cardiotocography signals. RESULTS: These accuracy metrics were calculated based on expert-labeled data. The algorithm achieved a classification accuracy of 96.2 % for acceleration, 94.4 % for deceleration, 90.9 % for tachycardia, and 85.8 % for bradycardia. Additionally, it achieved 67.0 % accuracy in classifying the four distinct deceleration patterns, with 80.9 % accuracy for late deceleration and 98.9 % for prolonged deceleration. CONCLUSIONS: The proposed multimodal deep learning algorithm serves as a reliable decision support tool for clinicians, significantly improving the detection and assessment of specific FHR events, which are crucial for fetal health monitoring.

Maximum Isokinetic Eccentric Elbow Flexor Muscle Force Can Be Estimated Using Maximum Isometric Contraction Force.

Purpose It is difficult to measure maximal isokinetic eccentric (ECC) muscle strength in the sports field. This study aimed to investigate whether elbow isometric (ISO) flexion muscle strength or muscle thickness (MT) can be used to estimate elbow ECC flexion muscle strength. Material and methods Maximal muscle strength and muscle thickness (MT) were measured in the elbow flexor muscle group of 147 healthy adults (age: 21.3±0.8 years, height: 167.3±8.6 cm, body mass: 61.4±10.6 kg: 99 males and 48 females). Both isometric contraction (ISO) and eccentric contraction (ECC) of elbow flexion muscle strength were measured using an isokinetic dynamometer. The ultrasound measured MT at 50% of the distance from the upper arm to the lateral epicondyle. We performed the multiple regression analysis with elbow ECC flexion muscle strength as the dependent variable and gender, age, height, body mass, elbow ISO flexion muscle strength, and MT as the independent variables. Results Multiple regression analysis revealed a coefficient of determination R2 value of 0.89 and an adjusted R2 value of 0.89 (p<0.01). In addition, the independent variables elbow ISO flexor strength (p<0.01, standardized coefficient ß=0.94; p<0.01, standardized coefficient ß=0.89) and muscle thickness (p<0.05, standardized coefficient ß=0.07) were identified as significantly associated factors. Conclusions The results suggest that it is possible to estimate elbow ECC flexion muscle strength using only elbow ISO flexion muscle strength and that ECC flexion muscle strength can be estimated more accurately by adding muscle thickness of the elbow flexor muscle group.

Long noncoding RNA VENTHEART is required for ventricular cardiomyocyte specification and function.

RATIONALE: Cardiac-expressed long noncoding RNAs (lncRNAs) are important for cardiomyocyte (CM) differentiation and function. Several lncRNAs have been identified and characterized for early CM lineage commitment, however those in later CM lineage specification and maturation remain less well studied. Moreover, unique atrial / ventricular lncRNA expression has never been studied in detail. OBJECTIVES: Here, we characterized a novel ventricular myocyte-restricted lncRNA, not expressed in atrial myocytes, and conserved only in primates. METHODS AND RESULTS: First, we performed single cell RNA-seq on human pluripotent stem cell derived cardiomyocytes (hPSC-CM) at the late stages of 2, 6 and 12 weeks of differentiation. Weighted correlation network analysis identified core gene modules, including a set of lncRNAs highly abundant and predominantly expressed in the human heart. A lncRNA (we call VENTHEART, VHRT) co-expressed with cardiac maturation and ventricular-specific genes MYL2 and MYH7, and was expressed in fetal and adult human ventricles, but not atria. CRISPR-mediated deletion of the VHRT gene led to impaired CM sarcomere formation and significant disruption of the ventricular CM gene program. Indeed, a similar disruption was not observed in VHRT KO hPSC-derived atrial CM, suggesting that VHRT exhibits only ventricular myocyte subtype-specific effects. Optical recordings validated that loss of VHRT significantly prolonged action potential duration at 90 % repolarization (APD90) for ventricular-like, but not atrial-like, CMs. CONCLUSION: This reports the first lncRNA that is exclusively required for proper ventricular, and not atrial, CM specification and function.

Mouse liver blood flow is regulated by hepatic stellate cells in response to the sympathetic neurotransmitter norepinephrine.

BACKGROUND: There is no clear information on the regulation of liver blood flow by the autonomic nervous system. We conducted this study to investigate whether quiescent hepatic stellate cells (qHSCs) regulate liver blood flow in response to the sympathetic neurotransmitter norepinephrine (NE). METHODS: qHSCs isolated from mice were cultured in Dulbecco's modified Eagle medium without fetal bovine serum for 1 day on collagen gel. NE-induced qHSC contraction was evaluated using the quantitative single-cell contraction measurement method that we had developed previously. For the measurement of liver perfusion pressure in situ, a buffer solution was perfused from the portal vein in mice. RESULTS: NE-induced a reversible contraction of qHSCs. This contraction was suppressed by the nonmuscle myosin II inhibitor blebbistatin, the myosin light chain kinase inhibitor ML-9, the Rho kinase inhibitor H-1152, the calmodulin inhibitor W-7, the store-operated calcium channel inhibitor YM-58483, and the IP3 receptor inhibitor xestospongin C. In contrast, the transient receptor potential C channel inhibitor SKF96365 did not affect the NE-induced contraction. CONCLUSION: These results suggest that qHSCs contract in response to NE. NEW & NOTEWORTHY: The present study provides direct evidence for the first time that norepinephrine (NE) induces a reversible contraction of isolated single quiescent hepatic stellate cells (qHSCs) and further suggests that the NE-mediated qHSC contraction participates in the regulation of liver blood flow in vivo.

Acute high-intensity muscle contraction moderates AChR gene expression independent of rapamycin-sensitive mTORC1 pathway in rat skeletal muscle.

The relationship between mechanistic target of rapamycin complex 1 (mTORC1) activation after resistance exercise and acetylcholine receptor (AChR) subunit gene expression remains largely unknown. Therefore, we aimed to investigate the effect of electrical stimulation-induced intense muscle contraction, which mimics acute resistance exercise, on the mRNA expression of AChR genes and the signalling pathways involved in neuromuscular junction (NMJ) maintenance, such as mTORC1 and muscle-specific kinase (MuSK). The gastrocnemius muscle of male adult Sprague-Dawley rats was isometrically exercised. Upon completion of muscle contraction, the rats were euthanized in the early (after 0, 1, 3, 6 or 24 h) and late (after 48 or 72 h) recovery phases and the gastrocnemius muscles were removed. Non-exercised control animals were euthanized in the basal state (control group). In the early recovery phase, Agrn gene expression increased whereas LRP4 decreased without any change in the protein and gene expression of AChR gene subunits. In the late recovery phase, Agrn, Musk, Chrnb1, Chrnd and Chrne gene expression were altered and agrin and MuSK protein expression increased. Moreover, mTORC1 and protein kinase B/Akt-histone deacetylase 4 (HDAC) were activated in the early phase but not in the late recovery phase. Furthermore, rapamycin, an inhibitor of mTORC1, did not disturb changes in AChR subunit gene expression after muscle contraction. However, rapamycin addition slightly increased AChR gene expression, while insulin did not impact it in rat L6 myotube. These results suggest that changes in the AChR subunits after muscle contraction are independent of the rapamycin-sensitive mTORC1 pathway.

Covariate-Assisted Bayesian Graph Learning for Heterogeneous Data.

In a traditional Gaussian graphical model, data homogeneity is routinely assumed with no extra variables affecting the conditional independence. In modern genomic datasets, there is an abundance of auxiliary information, which often gets under-utilized in determining the joint dependency structure. In this article, we consider a Bayesian approach to model undirected graphs underlying heterogeneous multivariate observations with additional assistance from covariates. Building on product partition models, we propose a novel covariate-dependent Gaussian graphical model that allows graphs to vary with covariates so that observations whose covariates are similar share a similar undirected graph. To efficiently embed Gaussian graphical models into our proposed framework, we explore both Gaussian likelihood and pseudo-likelihood functions. For Gaussian likelihood, a G-Wishart distribution is used as a natural conjugate prior, and for the pseudo-likelihood, a product of Gaussianconditionals is used. Moreover, the proposed model has large prior support and is flexible to approximate any v-Hölder conditional variance-covariance matrices with v ∈ ( 0,1 ] . We further show that based on the theory of fractional likelihood, the rate of posterior contraction is minimax optimal assuming the true density to be a Gaussian mixture with a known number of components. The efficacy of the approach is demonstrated via simulation studies and an analysis of a protein network for a breast cancer dataset assisted by mRNA gene expression as covariates.

Rotigaptide inhibits spontaneous contractions of gastric smooth muscle in diabetic rats via the PKCα-Cx43 pathway.

The study aimed to investigate the effect of rotigaptide (ZP123) on spontaneous contractions of gastric smooth muscle in diabetic rats and explore the underlying mechanisms. Twelve rats were randomly divided into model and normal control groups. Changes in gastric smooth muscle spontaneous contractions in each group were observed. Western blot analysis was performed to detect Cx43 and PKCα expression. Rat gastric smooth muscle cells were cultured in vitro and divided into normal glucose, high glucose and high glucose+rotigaptide group. The intracellular Ca2+ content was observed by immunofluorescence. The amplitude and frequency of gastric smooth muscle spontaneous contractions were reduced in the model group than the normal control group (all p < .01), which were reduced after rotigatide treatment than before treatment in the model group (all p < .01). The model+rotigaptide group showed decreased membrane expression of Cx43, increased cytoplasmic expression of Cx43, increased membrane expression of p-PKCα Thr497 and lower membrane/cytoplasm ratio of Cx43 expression compared with the model group (all p < .01). The intracellular Ca2+ content was increased in the high glucose group than the normal glucose group (p < .01), while no significant difference was observed between the high glucose+rotigaptide and high glucose groups. Our findings suggest that rotigatide can stabilize the intracellular Ca2+ concentration in gastric smooth muscle cells under high glucose condition by upregulating PKCα activity and downregulating the number of GJs and the opening rate of GJ hemichannels through the PKCα-Cx43 pathway, thus inhibiting spontaneous contractions of gastric smooth muscle in diabetic rats.

Direct Effects of Inflammatory Cytokines on Mouse Uterine Contraction.

PROBLEM: Uterine contractions signal labor onset, with elevated pro-inflammatory cytokines playing a pivotal role. Prior studies have explored their effects on prostaglandins, oxytocin, and signaling pathways, but have overlooked their direct effects on uterine contractions. Here, we aim to investigate the direct effects of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) on contractions to ascertain if they have immediate observable effects like those reported for lipopolysaccharide (LPS) and other effects. METHOD OF STUDY: Tension recordings were used to assess the direct effects of cytokines and/or LPS on mouse uterine contractions. Calcium imaging was employed to observe calcium oscillations in cytokine-pretreated myometrial smooth muscle cells (MSMCs) in response to oxytocin. The release of inflammatory cytokines and chemokines from uterine explants after LPS and/or cytokines application was investigated using Luminex. RESULTS: IL-1ß, IL-6, and TNF-α rapidly enhanced contractions of term pregnant mouse uterus. LPS combined with TNF-α intensified contractions compared to LPS alone, although this effect was not statistically significant in our results (p > 0.050). Pretreatment of MSMCs with IL-1ß, IL-6, or TNF-α increased calcium oscillations in response to oxytocin. LPS and/or cytokine significantly stimulated the release of IL-1ß, IL-6, TNF-α, Chemokine (C-X-C motif) ligand 1 (CXCL1), and monocyte chemoattractant protein-1 (MCP1) from uterine explants in vitro. CONCLUSIONS: Inflammatory cytokines have short-term and long-term effects on mouse uterine contractions, which together contribute to progressively stronger contractions during labor.

Ultrasound Assessment and Self-Perception of Pelvic Floor Muscle Function in Women with Stress Urinary Incontinence in Different Positions.

Objectives: This study analyzed the effect of different positions on pelvic floor muscle (PFM) function in women with and without stress urinary incontinence (SUI). Methods: This study included women with (n = 17, research group) and without (n = 25, control group) SUI. Using abdominal ultrasound, PFM function (maximum contraction and endurance) was measured in four different positions: lying, sitting, standing and squatting. The level of difficulty perceived by the participants was recorded. Results: In both groups, the best contraction was observed in the standing position and the weakest in the lying position. Women with SUI showed a lower ability to perform PFM contraction. A significant difference was found between the groups in the sitting and standing positions, and it was smaller in the research group. In the research group, the contraction displacement during sitting was 2.68 (1.67) mm versus 4.51 (2.62) mm in the control. The displacement during standing was 6.92 (3.50) mm versus 9.18 (5.05) mm, respectively (p < 0.05). In the research group, 52.9% reported the sitting position as the most difficult compared with only 12% in the control group. Conclusions: Women with SUI have lower PFM function while standing or sitting, but not while lying, than those without SUI. Variations in PFM function across different positions exist. A new protocol for PFM examination should be written with the standing position included.

Golden Jackal (Canis aureus) death by road accident in rural Bengal-insight the fact: a perception based eco-spatial clarification.

Golden jackal is one of the higher trophic level wild animals in rural Bengal. Their ecological significance is undeniable in regional ecosystems and biodiversity. But gradually they become threatened due to habitat contraction, road killing, poisoning and human persecution. The documentation of jackal death by road accident within the 12-km district highway which is a single lane bituminous road in Patshpur-I Community Development Block under Purba Medinipur district, India, reveals an unpredictable ecological circumstance (n-154). Jackals are killed by road accidents from 2022 to 2023. On behalf of this blameless situation, the present study conducted intensive observations, case studies and spatial-ecological analysis. The study also included stakeholder's perceptions to reach and relates the background of this fact. After observation and analysis, it has been found that the jackal deaths have a spatio-temporal signature and are connected with food and forage ground loss from their habitat-influenced area. Another outcome is that roadside habitat becomes an alternative food source and forage ground for them due to trench construction both alongside of the district highway and illegal dumping of meat wastes and domestic wastes. The seasonal biological activities of the jackals, local weather phenomena and car driver negligence are responsible for the accidental death of jackals along the district highway. Considering all the observed factors, the present study shared some reliable measures to control, protect and conserve this animal for balancing regional ecosystem as well as biodiversity in rural Bengal.

Engineering Shape to Overcome Contraction: The Role of Polymer-Collagen Hybrids in Advanced Dermal Substitutes.

Collagen gels are the standard dermal equivalents par excellence, however the problem of rapid cell-mediated contraction remains unresolved. Therefore, the development of hybrid constructs (HCs) based on collagen and polymeric scaffolds is proposed to address the mechanical instability that usually limits the formation of new, functional tissue. Equally important, these synthetic structures should be temporary (degradable) while ensuring that cells are well-adapted to the new extracellular environment. In this study, we screened a library of scaffolds made of various polymers, including homopolymers of polycaprolactone (PCL) and poly D,L-lactide (PLA50), their blends (PCL/PLA50), and copolymers (poly(D,L-lactide-co-caprolactone), PCLLA50) to prepare HCs in a layer-by-layer fashion. The properties of polymers and copolymers along with their processability by electrospinning and 3D-printing were evaluated. Then, we assessed the HCs resistance toward cell-mediated contraction as well as the degradation of the polymeric scaffolds. Our results indicate that scaffolds with higher PLA50 content (e.g., PLA50 100%, PCL/PLA50 or PCLLA50, both at 50/50 caprolactone-to-D,L-lactide molar ratio) presented more drawbacks in terms of handleability and processing, while those with greater PCL presence showed structural steadiness and ease to use. All the scaffolds integrated well with the collagen gel to form the corresponding HCs. With few exceptions, the HCs demonstrated good resistance to cell-derived contraction over 3 weeks. Notably, HCs based on PCLLA50 90/10 (both versions, electrospun or 3D-printed) performed best, showing only a 5%-17% area reduction compared to the 93% observed in collagen-only gels. This copolymer displayed hydrolytic degradation depending on its shape, with up to 45% and 65% loss of molecular weight for the electrospun and 3D-printed forms, respectively, correlating with their progressive change in mechanical features. HCs containing PCLLA50 90/10 also exhibited a better fibroblast distribution, enhanced myofibroblastic differentiation, and a three-fold increase in cell proliferation (when the electrospun type was used) compared to collagen controls. These findings were instrumental in selecting a potential HC that might be used for future experiments in vivo.

Shannon entropy variation as a global indicator of electron density contraction at interatomic regions in chemical reactions.

CONTEXT: The negative of the Shannon entropy derivative is proposed to account for electron density contraction as the chemical bonds are breaking and forming during a chemical reaction. We called this property the electron density contraction index, EDC, which allows identifying stages in a reaction that are dominated by electron contraction or expansion. Four different reactions were analyzed to show how the EDC index changes along the reaction coordinate. The results indicate that the rate of change of Shannon entropy is directly related to the rate of change of the electron density at the bond critical points between all the atomic pairs in the molecular systems. It is expected that EDC will complement the detailed analysis of reaction mechanisms that can be performed with the theoretical tools available to date. METHODS: Density functional theory calculations at the B3LYP/6-31G(d,p) level of theory were carried out using Gaussian 16 to analyze the reaction mechanisms of the four reactions studied. The reaction paths were obtained via the intrinsic reaction coordinate method, which served as the reaction coordinate to obtain the reaction force and the EDC profiles in each case. Shannon entropy and electron density at the bond critical points were calculated using the Multiwfn 3.7 package.

Native mechano-regulative matrix properties stabilize alternans dynamics and reduce spiral wave stabilization in cardiac tissue.

The stability of wave conduction in the heart is strongly related to the proper interplay between the electrophysiological activation and mechanical contraction of myocytes and extracellular matrix (ECM) properties. In this study, we statistically compare bioengineered cardiac tissues cultured on soft hydrogels ( E ≃ 12 kPa) and rigid glass substrates by focusing on the critical threshold of alternans, network-physiological tissue properties, and the formation of stable spiral waves that manifest after wave breakups. For the classification of wave dynamics, we use an improved signal oversampling technique and introduce simple probability maps to identify and visualize spatially concordant and discordant alternans as V- and X-shaped probability distributions. We found that cardiac tissues cultured on ECM-mimicking soft hydrogels show a lower variability of the calcium transient durations among cells in the tissue. This lowers the likelihood of forming stable spiral waves because of the larger dynamical range that tissues can be stably entrained with to form alternans and larger spatial spiral tip movement that increases the chance of self-termination on the tissue boundary. Conclusively, we show that a dysfunction in the excitation-contraction coupling dynamics facilitates life-threatening arrhythmic states such as spiral waves and, thus, highlights the importance of the network-physiological interplay between contractile myocytes and the ECM.

Investigating premotor corticospinal excitability in fast and slow voluntary contractions of the elbow flexors.

Corticospinal excitability (CSE) increases prior to a voluntary contraction; however, the relative contributions of premotor cortical and spinal mechanisms are poorly understood. It is unknown whether the intended voluntary contractile rate affects CSE. Eighteen young, healthy participants (nine females) completed isometric elbow flexion contractions targeting 50% maximal voluntary contraction (MVC) torque, at either fast (fast as possible) or slow (25% MVC/s) contractile rates. Participants were cued to contract with warning (red) and "GO" (green) visual signals. Magnetic and electric stimulations were applied to elicit motor evoked potentials (MEPs), cervicomedullary evoked potentials (CMEPs), and M-waves, in the surface electromyogram (EMG) recorded over the biceps brachii. MEPs and CMEPs were collected at 0, 25, 50 and 75% premotor reaction time (RT - defined as the time between the "GO" cue and onset of biceps brachii EMG) and compared to a resting baseline. MEP amplitude was greater than baseline at 75% RT (p=0.009), and CMEP amplitude was significantly increased at all RT points relative to baseline (p≤0.001). However, there were no differences in MEP and CMEP amplitudes when compared between fast and slow conditions (p≥0.097). Normalized to the CMEP, there was no difference in MEP amplitude from baseline in either contractile condition (p≥0.264). These results indicate that increased premotor CSE is a spinally-mediated response. Furthermore, premotor CSE is not influenced by the intended voluntary contractile rate. CMEP amplitudes were larger for females than males within the premotor RT period (p=0.038), demonstrating that premotor spinal excitability responses may be influenced by sex.

Excitation-contraction coupling inhibitors potentiate the actions of botulinum neurotoxin type A at the neuromuscular junction.

BACKGROUND AND PURPOSE: Botulinum neurotoxin type A1 (BoNT/A) is one of the most potent neurotoxins known. At the same time, it is also one of the safest therapeutic agents used for the treatment of several human disorders and in aesthetic medicine. Notwithstanding great effectiveness, strategies to accelerate the onset and prolong BoNT/A action would significantly ameliorate its pharmacological effects with beneficial outcomes for clinical use. EXPERIMENTAL APPROACH: Here, we combined BoNT/A with two fast-acting inhibitors of excitation-contraction coupling inhibitors (ECCI), either the µ-conotoxin CnIIIC or dantrolene, and tested the effect of their co-injection on a model of hind-limb paralysis in rodents using behavioural, biochemical, imaging and electrophysiological assays. KEY RESULTS: The BoNT/A-ECCI combinations accelerated the onset of muscle relaxation. Surprisingly, they also potentiated the peak effect and extended the duration of the three BoNT/A commercial preparations OnabotulinumtoxinA, AbobotulinumtoxinA and IncobotulinumtoxinA. ECCI co-injection increased the number of BoNT/A molecules entering motoneuron terminals, which induced a faster and greater cleavage of SNAP-25 during the onset and peak phases, and prolonged the attenuation of nerve-muscle neurotransmission during the recovery phase. We estimate that ECCI co-injection yields a threefold potentiation in BoNT/A pharmacological activity. CONCLUSIONS AND IMPLICATIONS: Overall, our results show that the pharmacological activity of BoNT/A can be combined and synergized with other bioactive molecules and uncover a novel strategy to enhance the neuromuscular effects of BoNT/A without altering the neurotoxin moiety or intrinsic activity, thus maintaining its exceptional safety profile.

Reduced K+ build-up in t-tubules contributes to resistance of the diaphragm to myotonia.

Patients with myotonia congenita suffer from slowed muscle relaxation caused by hyperexcitability. The diaphragm is only mildly affected in myotonia congenita; discovery of the mechanism underlying its resistance to myotonia could identify novel therapeutic targets. Intracellular recordings from two mouse models of myotonia congenita revealed the diaphragm had less myotonia than either the extensor digitorum longus (EDL) or the soleus muscles. A mechanism contributing to resistance of the diaphragm to myotonia was reduced depolarization of the interspike membrane potential during repetitive firing of action potentials, a process driven by build-up of K+ in small invaginations of muscle membrane known as t-tubules. We explored differences between diaphragm and EDL that might underlie reduction of K+ build-up in diaphragm t-tubules. Smaller size of diaphragm fibres, which promotes diffusion of K+ out of t-tubules, was identified as a contributor. Intracellular recording revealed slower repolarization of action potentials in diaphragm suggesting reduced Kv conductance. Higher resting membrane conductance was identified suggesting increased Kir conductance. Computer simulation found that a reduction of Kv conductance had little effect on K+ build-up whereas increased Kir conductance lessened build-up, although the effect was modest. Our data and computer simulation suggest opening of K+ channels during action potentials has little effect on K+ build-up whereas opening of K+ channels during the interspike interval slightly lessens K+ build-up. We conclude that activation of K+ channels may lessen myotonia by opposing depolarization to action potential threshold without worsening K+ build-up in t-tubules. KEY POINTS: In mouse models of the muscle disease myotonia congenita, the diaphragm has much less myotonia (muscle stiffness) than the extensor digitorum longus or soleus muscles. Identifying why the diaphragm is resistant to myotonia may help in developing novel therapy. We found the reason the diaphragm has less myotonia is that it is less prone to depolarization caused by K+ build-up in t-tubules during repetitive firing of action potentials. Smaller fibre size contributes to resistance to K+ build-up with differences in K+ currents playing little role. Our data suggest drugs that open K+ channels may be effective in treating myotonia as they may lessen excitability without worsening K+ build-up in t-tubules.

PRP improves the outcomes of autologous skin graft transplantation on the esophagus by promoting angiogenesis and inhibiting fibrosis and inflammation.

Background and Objectives: Autologous skin graft (ASG) transplantation is a challenging approach but a promising option for patients to prevent postoperative esophageal stricture. Nonetheless, the current strategies require improvement. We aimed to investigate the effectiveness of the injection of platelet-rich plasma (PRP) before skin graft transplantation for extensive esophageal defects after endoscopic resection. Methods: Standardized complete circular endoscopic resection (5 cm in length) was performed in 27 pigs allocated into 3 groups. The artificial ulcers were treated with a fully covered esophageal stent (control group), ASG (ASG group), and submucosal injection of PRP with ASG (PRP-ASG group). Macroscopic evaluation and histological analysis of the remolded esophagus were performed 7, 14, and 28 days after surgery. Results: The macroscopic evaluation indicated that submucosal injection of PRP before transplantation effectively promoted the survival rate of skin grafts and decreased the rate of mucosal contraction compared with those treated with ASG or stent alone. Histological analysis of submucosal tissue showed that this modified strategy significantly promoted wound healing of reconstructed tissues by enhancing angiogenesis, facilitating collagen deposition, and decreasing inflammation and fibrogenesis. Conclusions: These findings suggested that PRP might be used as a biological supplement to increase the esophageal skin graft survival rate and improve submucosal tissue remolding in a clinically relevant porcine model. With extremely low mucosal contraction, this novel combination strategy showed the potential to effectively prevent stenosis in extensive esophageal ulcers.