Relationship Between Resultant Force Vector Acting on Human Organs From Food Bolus and the Bolus Configuration During Swallowing Using Numerical Swallowing Simulation With Moving Particle Simulation Method.

This study investigates the forces exerted on organs during swallowing, specifically focusing on identifying forces other than those resulting from direct organ contact. Using a swallowing simulator based on the moving particle method, we simulated the swallowing process of healthy individuals upon the ingestion of thickened foods, which were simulated as shear-thinning flow without yield stress. We extracted the resultant force vectors acting on the organs and shape of the bolus at each time interval. The simulation results confirmed that the bolus originates from tongue movement and is transferred between the oral cavity and pharynx, with each organ's coordinated movements with the tongue occurring at their respective positions, as indicated by the balance of the resultant force vectors. Utilizing the information about the resultant force vectors obtained through simulations, we calculated the physical parameters of impulse, energy, and power. The variations in these physical parameters were aligned with the behaviors of both the biological system and the food bolus during swallowing. The force values calculated from the simulations closely approximate the theoretical values. Furthermore, the forces calculated from the simulations were relatively smaller than the force values derived from pressure information, such as that from high-resolution manometry and tongue pressure sensors. This difference can be attributed to the simulations extracting only the forces exerted on the organ by the food bolus. Force information on organs has the potential to provide a new interpretation of conventional mechanical indicators such as manometry and tongue pressure sensors.

Standardization of Stimulus Location for Functional Electrical Stimulation of Swallowing.

Many nerves and muscles are involved in the swallowing process; hence neuromuscular disorders cause dysphagia resulting in aspiration pneumonia. A critical movement in the pharyngeal phase of swallowing is hyolaryngeal elevation to help protect the airway and open a relaxed upper esophageal sphincter. Functional electrical stimulation (FES) is expected to improve the function of muscles acting on the hyolaryngeal motion, which may contribute to airway protection in dysphagic patients. However, it is difficult to select the stimulus locations that effectively assist laryngeal elevation without the expertise in the anatomy of swallowing-related muscles. Therefore, this study investigated the method to standardize the selection of the stimulus locations based on the dimensions of the larynx. In addition, the effect of stimulus intensity on the amount of laryngeal elevation was evaluated.

Recovery of motor function of chronic spinal cord injury by extracellular pyruvate kinase isoform M2 and the underlying mechanism.

In our previous study, we found that pyruvate kinase isoform M2 (PKM2) was secreted from the skeletal muscle and extended axons in the cultured neuron. Indirect evidence suggested that secreted PKM2 might relate to the recovery of motor function in spinal cord injured (SCI) mice. However, in vivo direct evidence has not been obtained, showing that extracellular PKM2 improved axonal density and motor function in SCI mice. In addition, the signal pathway of extracellular PKM2 underlying the increase in axons remained unknown. Therefore, this study aimed to identify a target molecule of extracellular PKM2 in neurons and investigate the critical involvement of extracellular PKM2 in functional recovery in the chronic phase of SCI. Recombinant PKM2 infusion to the lateral ventricle recovered motor function in the chronic phase of SCI mice. The improvement of motor function was associated with axonal increase, at least of raphespinal tracts connecting to the motor neurons directly or indirectly. Target molecules of extracellular PKM2 in neurons were identified as valosin-containing protein (VCP) by the drug affinity responsive target stability method. ATPase activation of VCP mediated the PKM2-induced axonal increase and recovery of motor function in chronic SCI related to the increase in axonal density. It is a novel finding that axonal increase and motor recovery are mediated by extracellular PKM2-VCP-driven ATPase activity.

Generation of mouse iPS cells using an inducible expression of transgenes via the cumate gene-switch.

We applied an inducible gene expression system that utilizes the p-cmt operon, the cumate gene-switch, to generate mouse induced pluripotent stem (iPS) cells. Mouse embryonic fibroblast (MEF) E6E7-MEF cells were transfected with a single cumate gene-switch vector enabling concomitant expression of Oct4, Sox2, c-Myc, Klf4, and Gfp. Then, the cells were cultured with cumate, a monoterpene. An increase in colonies positive for alkaline phosphatase activity was observed dose-dependently with cumate. In the absence of cumate, the expression of GFP, a marker for transgene expression, was undetectable in tightly aggregated iPS cell-like colonies with endogenous expression of NANOG and OCT4. From primary MEFs using the cumate gene-switch, we also isolated iPS cells expressing endogenous NANOG, OCT4, SOX2, KLF4, and SSEA1 with hypo-methylated genomic promoter regions of endogenous Nanog and Oct4. In embryoid bodies with the progression of differentiation, expression of markers for all three germ layers was detected, and contracting cardiomyocytes were observed. Overall, we suggest that the cumate gene-switch is applicable for the generation of mouse iPS cells. The cumate gene-switch in combination with other inducible systems, such as the tet system, may provide useful approaches for analyzing the roles of transgenes underlying the establishment of iPS cells.

Estimation of Muscle Activity Change under Different Bolus Conditions using Musculoskeletal Model of Swallowing.

Swallowing, deglutition, is realized by highly coordinated activities of many nerves and muscles, but it is hard to observe directly due to intracorporal movement, and there is a limitation to the number of muscles that can be percutaneously measured. In addition, since there are few studies on the mechanical analysis of the swallowing movement, the detailed muscle activity pattern during swallowing has not yet been clarified. To tackle this problem from the viewpoint of biomechanics, we have been developing the musculoskeletal model of swallowing which can estimate the activities of swallowing-related muscles based on the movements of hyoid bone and thyroid cartilage. In this paper, we analyzed the activities of swallowing-related muscles under two different bolus conditions: bolus of water and nectar thickened liquid to investigate the effect of physical property of bolus in the activities of swallowing-related muscles.

Acteoside Improves Muscle Atrophy and Motor Function by Inducing New Myokine Secretion in Chronic Spinal Cord Injury.

Chronic spinal cord injury (SCI) is difficult to cure, even by several approaches effective at the acute or subacute phase. We focused on skeletal muscle atrophy as a detrimental factor in chronic SCI and explored drugs that protect against muscle atrophy and activate secretion of axonal growth factors from skeletal muscle. We found that acteoside induced the secretion of axonal growth factors from skeletal muscle cells and proliferation of these cells. Intramuscular injection of acteoside in mice with chronic SCI recovered skeletal muscle weight reduction and motor function impairment. We also identified pyruvate kinase isoform M2 (PKM2) as a secreted factor from skeletal muscle cells, stimulated by acteoside. Extracellular PKM2 enhanced proliferation of skeletal muscle cells and axonal growth in cultured neurons. Further, we showed that PKM2 might cross the blood-brain barrier. These results indicate that effects of acteoside on chronic SCI might be mediated by PKM2 secretion from skeletal muscles. This study proposes that the candidate drug acteoside and a new myokine, PKM2, could be used for the treatment of chronic SCI.

Unified Total Synthesis, Stereostructural Elucidation, and Biological Evaluation of Sarcophytonolides.

Sarcophytonolides are cembranolide diterpenes isolated from the soft corals of genus Sarcophyton. Unified total synthesis of sarcophytonolides C, E, F, G, H, and J and isosarcophytonolide D was achieved. The synthetic routes feature NaHMDS- or SmI2-mediated fragment coupling, alkoxycarbonylallylation, macrolactonization, and transannular ring-closing metathesis. These total syntheses led to the absolute configurational confirmation of sarcophytonolide H, elucidation of sarcophytonolides C, E, F, and G, and revision of sarcophytonolide J and isosarcophytonolide D. We also evaluated the antifouling activity and toxicity of the synthetic sarcophytonolides H and J and their analogues as well as the cytotoxicity of the synthetic sarcophytonolides and the key synthetic intermediates.

Late-stage divergent synthesis and antifouling activity of geraniol-butenolide hybrid molecules.

Hybrid molecules consisting of geraniol and butenolide were designed and synthesized by the late-stage divergent strategy. In the synthetic route, ring-closing metathesis was utilized for the construction of a butenolide moiety. A biological evaluation of the eight synthetic hybrid compounds revealed that these molecules exhibit antifouling activity against the cypris larvae of the barnacle Balanus (Amphibalanus) amphitrite with EC50 values of 0.30-1.31 µg mL-1. These results show that hybridization of the geraniol and butenolide structural motifs resulted in the enhancement of the antifouling activity.

Numerical simulation of interaction between organs and food bolus during swallowing and aspiration.

The mechanism of swallowing is still not fully understood, because the process of swallowing is a rapid and complex interaction among several involved organs and the food bolus. In this work, with the aim of studying swallowing and aspiration processes noninvasively and systematically, a computer simulation method for analyzing the involved organs and water (considered as the food bolus) is proposed. The shape and motion of the organs involved in swallowing are modeled in the same way as in our previous study, by using the Hamiltonian moving particle simulation (MPS) method and forced displacements on the basis of motion in a healthy volunteer. The bolus flow is simulated using the explicit MPS method for fluid analysis. The interaction between the organs and the bolus is analyzed using a fluid-structure coupling scheme. To validate the proposed method, the behavior of the simulated bolus flow is compared qualitatively and quantitatively with corresponding medical images. In addition to the healthy motion model, disorder motion models are constructed for reproducing the aspiration phenomenon by computer simulation. The behaviors of the organs and the bolus considered as the food bolus in the healthy and disorder motion models are compared for evaluating the mechanism of aspiration.

Knockdown of gene expression by antisense morpholino oligos in preimplantation mouse embryos cultured in vitro.

Knockdown of gene expression by antisense morpholino oligos (MOs) is a simple and effective method for analyzing the roles of genes in mammalian cells. Here, we demonstrate the efficient delivery of MOs by Endo-Porter (EP), a special transfection reagent for MOs, into preimplantation mouse embryos cultured in vitro. A fluorescein-labeled control MO was applied for monitoring the incorporation of MOs into developing 2-cell embryos in the presence of varying amounts of EP and bovine serum albumin. In optimized conditions, fluorescence was detected in 2-cell embryos within a 3-h incubation period. In order to analyze the validity of the optimized conditions, an antisense Oct4 MO was applied for knockdown of the synthesis of OCT4 protein in developing embryos from the 2-cell stage. In blastocysts, the antisense Oct4 MO induced a decrease in the amount in OCT4 protein to less than half. An almost complete absence of OCT4-positive cells and nearly complete disappearance of the inner cell mass in the outgrowths of blastocysts were also noted. These phenotypes corresponded with those of Oct4-deficient mouse embryos. Overall, we suggest that the delivery of MOs using EP is useful for the knockdown of gene expression in preimplantation mouse embryos cultured in vitro.