Impact of moderate-intensity aerobic exercise in combined hypoxic and hot conditions on endothelial function.

There is no study that has investigated the impact of exercise in a combined hypoxic and hot environment on endothelial function. Therefore, we tested whether aerobic exercise in a combined hypoxic and hot conditions induces further enhancement of endothelial function. Twelve healthy males cycled at a constant workload (50% of their maximal oxygen uptake under normoxic/thermoneutral conditions) for 30 min in four different environments: exercise under normoxic condition (NOR: fraction of inspiratory oxygen or FiO2 = 20.9%, 20°C), exercise under hypoxic condition (HYP: FiO2 = 14.5%, 20°C), exercise under hot condition (HOT: FiO2 = 20.9%, 30°C), and exercise under combined hypoxia and hot conditions (HH: FiO2 = 14.5%, 30°C). Before, during, and after exercise, cardiovascular variables (e.g., heart rate, blood flow, and shear rate), blood variables, and endothelial function evaluated by flow-mediated dilation (FMD) were assessed. Heart rates were significantly higher throughout the HH trial's experimental period than the other trials (p < 0.05). However, in the HH trial, brachial artery blood flow and shear rate did not differ from those in other trials after exercise. Plasma catecholamines (epinephrine, norepinephrine, and dopamine) elevations in response to exercise were significantly higher in the HH trial than in the other three trials (p < 0.05). No considerable differences were observed in FMD responses among trials before and after the exercise. In conclusion, aerobic exercise in a combined hot and hypoxic environment further activated sympathetic nervous activity but did not considerably enhance blood flow, shear rate, or endothelial function.

[New Development in Amyloidosis Research Based on Supersaturation: Biological Factors to Induce/Inhibit the Amyloid Fibril Formation].

Amyloid fibril formation is a general property of proteins and peptides. It is a physicochemical phenomenon similar to crystallization, in which amyloid precursor proteins exceeding solubility precipitate through the breakdown of supersaturation. Using the ultrasonication-forced amyloid fibril inducer HANABI, we have discovered that serum albumin acts as an inhibitor in dialysis-related amyloidosis. Exploring the factors that induce or inhibit amyloid fibril formation using HANABI can lead to the development of early diagnosis and prevention methods for amyloidosis.

Surface Modification of Ultrasonic Cavitation by Surfactants Improves Detection Sensitivity of α-Synuclein Amyloid Seeds.

The rapid amplification and sensitive detection of α-synuclein (αSyn) seeds is an efficient approach for the early diagnosis of Parkinson's disease. Ultrasonication stands out as a promising method for the rapid amplification of αSyn seeds because of its robust fibril fragmentation capability. However, ultrasonication also induces the primary nucleation of αSyn monomers, deteriorating the seed detection sensitivity by generating seed-independent fibrils. In this study, we show that an addition of surfactants to the αSyn monomer solution during αSyn seed detection under ultrasonication remarkably improves the detection sensitivity of the αSyn seeds by a factor of 100-1000. Chemical kinetic analysis reveals that these surfactants reduce the rate of primary nucleation while promoting the fragmentation of the αSyn fibrils under ultrasonication. These effects are attributed to the modification of the ultrasonic cavitation surface by the surfactants. Our study enhances the utility of ultrasonication in clinical assays targeting αSyn seeds as the Parkinson's disease biomarker.

Supersaturation, a Critical Factor Underlying Proteostasis of Amyloid Fibril Formation.

From a physicochemical viewpoint, amyloid fibril formation is a phase transition from soluble to crystal-like sates limited by supersaturation. It occurs only above solubility (i.e., the solubility limit) coupled with a breakdown of supersaturation. Although many studies have examined the role of molecular chaperones in the context of proteostasis, the role of supersaturation has not been addressed. Moreover, although molecular chaperone-dependent disaggregations have been reported for preformed amyloid fibrils, amyloid fibrils will not dissolve above the solubility of monomers, even if agitations fragment long fibrils to shorter amyloid particles. On the other hand, on considering a reversible and coupled equilibrium of interactions, folding/unfolding and amyloid formation/disaggregation, molecules stabilizing native states can work as a disaggregase reversing the amyloid fibrils to monomers. It is likely that the proteostasis network has various intra- and extracellular components which disaggregate preformed amyloid fibrils as well as prevent amyloid formation. Further studies with a view of solubility and supersaturation will be essential for comprehensive understanding of proteostasis.

Mass spectrometry-based proteomic analysis of proteins adsorbed by hexadecyl-immobilized cellulose bead column for the treatment of dialysis-related amyloidosis.

BACKGROUND: Dialysis-related amyloidosis (DRA) is a severe complication in end-stage kidney disease (ESKD) patients undergoing long-term dialysis treatment, characterized by the deposition of ß2-microglobulin-related amyloids (Aß2M amyloid). To inhibit DRA progression, hexadecyl-immobilized cellulose bead (HICB) columns are employed to adsorb circulating ß2-microglobulin (ß2M). However, it is possible that the HICB also adsorbs other molecules involved in amyloidogenesis. METHODS: We enrolled 14 ESKD patients using HICB columns for DRA treatment; proteins were extracted from HICBs following treatment and identified using liquid chromatography-linked mass spectrometry. We measured the removal rate of these proteins and examined the effect of those molecules on Aß2M amyloid fibril formation in vitro. RESULTS: We identified 200 proteins adsorbed by HICBs. Of these, 21 were also detected in the amyloid deposits in the carpal tunnels of patients with DRA. After passing through the HICB column and hemodialyzer, the serum levels of proteins such as ß2M, lysozyme, angiogenin, complement factor D and matrix Gla protein were reduced. These proteins acted in the Aß2M amyloid fibril formation. CONCLUSIONS: HICBs adsorbed diverse proteins in ESKD patients with DRA, including those detected in amyloid lesions. Direct hemoperfusion utilizing HICBs may play a role in acting Aß2M amyloidogenesis by reducing the amyloid-related proteins.

Sodium L-Aspartate Supplementation Improves Repeated-Sprint Performance.

Aspartate supplementation has been reported to improve endurance performance by facilitating the tricarboxylic acid cycle flux. The present study was performed to investigate the effects of aspartate supplementation on repeated-sprint performance and blood pH. Following an overnight fast, fourteen healthy males completed three sets of 10 × 6 s maximal sprints after consuming sodium L-aspartate (ASP) or placebo (PLA), in a double-blind manner. Both supplements were taken twice on each test day (2 × 4.5 g). Exercise performance (e.g., cadence and power output) and blood variables (e.g., pH and plasma amino acid levels) were measured. The ASP trial evidenced significantly higher plasma aspartate concentration during the first (ASP, 45.3 ± 9.2 µM; PLA, 6.1 ± 0.8 µM) and the second exercise sets (ASP, 24.2 ± 4.5 µM; PLA, 6.6 ± 0.9 µM) and peak cadence during the second set (ASP, 153 ± 3 rpm; PLA, 152 ± 3 rpm) compared with the PLA trial (all p < 0.05). The peak power output during the second exercise set (ASP, 743 ± 32 W; PLA, 734 ± 31 W; p = 0.060) and the blood pH immediately before (ASP, 7.280 ± 0.020; PLA, 7.248 ± 0.016; p = 0.087) and after the third exercise set (ASP, 7.274 ± 0.019; PLA, 7.242 ± 0.018; p = 0.093) tended to be higher in the ASP than in the PLA trial. In conclusion, ASP supplementation partially improved repeated-sprint performance (peak cadence during the second exercise set). However, it did not affect the mean power output.

Mechanisms of polyphosphate-induced amyloid fibril formation triggered by breakdown of supersaturation.

Much effort has been devoted to elucidate mechanisms of amyloid fibril formation using various kinds of additives, such as salts, metals, detergents, and biopolymers. Here, we review the effects of additives with a focus on polyphosphate (polyP) on amyloid fibril formation of ß2-microglobulin (ß2m) and α-synuclein (αSyn). PolyP, consisting of up to 1,000 phosphoanhydride bond-linked phosphate monomers, is one of the most ancient, enigmatic, and negatively charged molecules in biology. Amyloid fibril formation of both ß2m and αSyn could be accelerated by counter anion-binding and preferential hydration at relatively lower and higher concentrations of polyP, respectively, depending on the chain length of polyP. These bimodal concentration-dependent effects were also observed in salt- and heparin-induced amyloid fibril formation, indicating the generality of bimodal effects. We also address the effects of detergents, alcohols, and isoelectric point precipitation on amyloid fibril formation, in comparison with the effects of salts. Because polyP is present all around us, from cellular components to food additives, clarifying its effects and consequent biological roles will be important to further advance our understanding of amyloid fibrils. This review article is an extended version of the Japanese article, Linking Protein Folding to Amyloid Formation, published in SEIBUTSU BUTSURI Vol. 61, p. 358-365 (2021).

Ultrastiff Amyloid-Fibril Network of α-Synuclein Formed by Surface Seeding Reaction Confirmed by Multichannel Electrodeless Quartz-Crystal-Microbalance Biosensor.

We developed a multichannel wireless quartz-crystal-microbalance (QCM) biosensor for mechanically studying the on-surface aggregation reaction of α-synuclein (α-syn). We find a quite unusual change in the resonant frequency that eventually exceeds the baseline, which has never been observed during seeding aggregation reaction. By incorporating a growth-to-percolation theory for fibril elongation reaction, we have favorably reproduced this unusual response and found that it can be explained only with formation of an ultrastiff fibril network. We also find that the stiffness of the fibril network grown from artificially prepared twist-type seeds is significantly higher than that from rod-type seeds. Furthermore, the stiffnesses of fibril networks grown from seeds derived from brain tissues of Parkinson's disease (PD) and multiple system atrophy (MSA) patients show a very similar trend to those of rod and twist seeds, respectively, indicating that fibrils from MSA patients are stiffer than those from PD.

Phosphatidylinositol-3,4,5-trisphosphate interacts with alpha-synuclein and initiates its aggregation and formation of Parkinson's disease-related fibril polymorphism.

Lipid interaction with α-synuclein (αSyn) has been long implicated in the pathogenesis of Parkinson's disease (PD). However, it has not been fully determined which lipids are involved in the initiation of αSyn aggregation in PD. Here exploiting genetic understanding associating the loss-of-function mutation in Synaptojanin 1 (SYNJ1), a phosphoinositide phosphatase, with familial PD and analysis of postmortem PD brains, we identified a novel lipid molecule involved in the toxic conversion of αSyn and its relation to PD. We first established a SYNJ1 knockout cell model and found SYNJ1 depletion increases the accumulation of pathological αSyn. Lipidomic analysis revealed SYNJ1 depletion elevates the level of its substrate phosphatidylinositol-3,4,5-trisphosphate (PIP3). We then employed Caenorhabditis elegans model to examine the effect of SYNJ1 defect on the neurotoxicity of αSyn. Mutations in SYNJ1 accelerated the accumulation of αSyn aggregation and induced locomotory defects in the nematodes. These results indicate that functional loss of SYNJ1 promotes the pathological aggregation of αSyn via the dysregulation of its substrate PIP3, leading to the aggravation of αSyn-mediated neurodegeneration. Treatment of cultured cell line and primary neurons with PIP3 itself or with PIP3 phosphatase inhibitor resulted in intracellular formation of αSyn inclusions. Indeed, in vitro protein-lipid overlay assay validated that phosphoinositides, especially PIP3, strongly interact with αSyn. Furthermore, the aggregation assay revealed that PIP3 not only accelerates the fibrillation of αSyn, but also induces the formation of fibrils sharing conformational and biochemical characteristics similar to the fibrils amplified from the brains of PD patients. Notably, the immunohistochemical and lipidomic analyses on postmortem brain of patients with sporadic PD showed increased PIP3 level and its colocalization with αSyn. Taken together, PIP3 dysregulation promotes the pathological aggregation of αSyn and increases the risk of developing PD, and PIP3 represents a potent target for intervention in PD.

Adding heat stress to repeated-sprint training in hypoxia does not enhance performance improvements in canoe/kayak athletes.

PURPOSE: The present study investigated the effects of adding heat stress to repeated-sprint training in hypoxia on performance and physiological adaptations in well-trained athletes. METHODS: Sixteen canoe/kayak sprinters conducted 2 weeks of repeated-sprint training consisting of three sets of 5 × 10 s sprints with 20 s active recovery periods under conditions of either normobaric hypoxia (RSH, FiO2: 14.5%, ambient temperature: 18 ℃, n = 8) or combined heat and normobaric hypoxia (RSHH, FiO2: 14.5%, ambient temperature: 38 ℃, n = 8). Before and after training, the 10 × 10 s repeated-sprint ability (RSA) test and 500 m time trial were performed on a canoe/kayak ergometer. RESULTS: Peak and average power outputs during the RSA test were significantly improved after training in both RSH (peak power: + 21.5 ± 4.6%, P < 0.001; average power: + 12.5 ± 1.9%, P < 0.001) and RSHH groups (peak power: + 18.8 ± 6.6%, P = 0.005; average power: + 10.9 ± 6.8%, P = 0.030). Indirect variables of skeletal muscle oxygen extraction (deoxygenated hemoglobin) and blood perfusion (total hemoglobin) during the RSA test were significantly increased after training in the RSH group (P = 0.041 and P = 0.034, respectively) but not in the RSHH group. In addition, finish time during the 500 m time trial was significantly shortened after the training only in the RSH group (RSH: - 3.9 ± 0.8%, P = 0.005; RSHH: - 3.1 ± 1.4%, P = 0.078). CONCLUSION: Adding heat stress to RSH does not enhance performance improvement and may partially mask muscle tissue adaptation.

The residual structure of acid-denatured ß2 -microglobulin is relevant to an ordered fibril morphology.

ß2 -Microglobulin (ß2m) forms amyloid fibrils in vitro under acidic conditions. Under these conditions, the residual structure of acid-denatured ß2m is relevant to seeding and fibril extension processes. Disulfide (SS) bond-oxidized ß2m has been shown to form rigid, ordered fibrils, whereas SS bond-reduced ß2m forms curvy, less-ordered fibrils. These findings suggest that the presence of an SS bond affects the residual structure of the monomer, which subsequently influences the fibril morphology. To clarify this process, we herein performed NMR experiments. The results obtained revealed that oxidized ß2m contained a residual structure throughout the molecule, including the N- and C-termini, whereas the residual structure of the reduced form was localized and other regions had a random coil structure. The range of the residual structure in the oxidized form was wider than that of the fibril core. These results indicate that acid-denatured ß2m has variable conformations. Most conformations in the ensemble cannot participate in fibril formation because their core residues are hidden by residual structures. However, when hydrophobic residues are exposed, polypeptides competently form an ordered fibril. This conformational selection phase may be needed for the ordered assembly of amyloid fibrils.

Macromolecular crowding and supersaturation protect hemodialysis patients from the onset of dialysis-related amyloidosis.

Dialysis-related amyloidosis (DRA), a serious complication among long-term hemodialysis patients, is caused by amyloid fibrils of ß2-microglobulin (ß2m). Although high serum ß2m levels and a long dialysis vintage are the primary and secondary risk factors for the onset of DRA, respectively, patients with these do not always develop DRA, indicating that there are additional risk factors. To clarify these unknown factors, we investigate the effects of human sera on ß2m amyloid fibril formation, revealing that sera markedly inhibit amyloid fibril formation. Results from over 100 sera indicate that, although the inhibitory effects of sera deteriorate in long-term dialysis patients, they are ameliorated by maintenance dialysis treatments in the short term. Serum albumin prevents amyloid fibril formation based on macromolecular crowding effects, and decreased serum albumin concentration in dialysis patients is a tertiary risk factor for the onset of DRA. We construct a theoretical model assuming cumulative effects of the three risk factors, suggesting the importance of monitoring temporary and accumulated risks to prevent the development of amyloidosis, which occurs based on supersaturation-limited amyloid fibril formation in a crowded milieu.

Comparison of systemic and peripheral responses during high-intensity interval exercise under voluntary hypoventilation vs. hypoxic conditions.

PURPOSE: This study aimed to determine the systemic and peripheral responses to high-intensity interval exercise (HIIE) with voluntary hypoventilation at low lung volume (VHL) or HIIE under hypoxic conditions. METHODS: Ten male participants completed a single session of HIIE (three sets of 6 × 8-s high-intensity pedaling at 170% of maximal oxygen uptake [VO2max]) under three different conditions: normoxia with normal breathing (NOR: 23 °C, 20.9% of fraction of inspired oxygen [FiO2]), hypoxia with normal breathing (HYP: 23 °C, 14.5% FiO2), and normoxia with VHL (VHL: 23 °C, 20.9% FiO2). A randomized crossover design was used. Power output, arterial oxygen saturation (SpO2), heart rate, and muscle oxygenation were monitored during the exercise and the 16-s recovery. Muscle blood flow (mBF) of the vastus lateralis was also evaluated. RESULTS: SpO2 during the exercise and the 16-s recovery in the VHL group was significantly lower than in that of the NOR group. However, this parameter in the VHL group was significantly higher than that of the HYP group (NOR: 94.9 ± 0.4%, HYP: 82.8 ± 1.2%, VHL: 90.4 ± 0.5%; p < 0.001). Muscle oxygen saturation was significantly lower in the HYP group than those in the VHL and NOR groups (NOR: 79.6 ± 17.4%, HYP: 65.5 ± 7.7%, VHL: 74.4 ± 7.8%; p = 0.024). No significant difference in this parameter was observed between the VHL and NOR groups (p > 0.05). Additionally, the exercise-induced increase in mBF did not differ significantly among three groups (p > 0.05). CONCLUSION: HIIE-induced SpO2 decrease was smaller under hypoxic conditions than during VHL. Moreover, mBF was not enhanced by the addition of VHL during HIIE.

Supersaturation-Dependent Formation of Amyloid Fibrils.

The supersaturation of a solution refers to a non-equilibrium phase in which the solution is trapped in a soluble state, even though the solute's concentration is greater than its thermodynamic solubility. Upon breaking supersaturation, crystals form and the concentration of the solute decreases to its thermodynamic solubility. Soon after the discovery of the prion phenomena, it was recognized that prion disease transmission and propagation share some similarities with the process of crystallization. Subsequent studies exploring the structural and functional association between amyloid fibrils and amyloidoses solidified this paradigm. However, recent studies have not necessarily focused on supersaturation, possibly because of marked advancements in structural studies clarifying the atomic structures of amyloid fibrils. On the other hand, there is increasing evidence that supersaturation plays a critical role in the formation of amyloid fibrils and the onset of amyloidosis. Here, we review the recent evidence that supersaturation plays a role in linking unfolding/folding and amyloid fibril formation. We also introduce the HANABI (HANdai Amyloid Burst Inducer) system, which enables high-throughput analysis of amyloid fibril formation by the ultrasonication-triggered breakdown of supersaturation. In addition to structural studies, studies based on solubility and supersaturation are essential both to developing a comprehensive understanding of amyloid fibrils and their roles in amyloidosis, and to developing therapeutic strategies.

Comparison of energy expenditure and substrate oxidation between walking and running in men and women.

PURPOSE: The present study compared energy metabolism between walking and running at equivalent speeds during two incremental exercise tests. METHODS: Thirty four university students (18 males, 16 females) were recruited. Each participant completed two trials, consisting of walking (Walk) and running (Run) trials on different days, with 2-3 days apart. Exercise on a treadmill was started from initial stage of 3 min (3.0 k/m in Walk trial, 5.0 km/h in Run trial), and the speed for walking and running was progressively every minute by 0.5 km/h. The changes in metabolic variables, heart rate (HR), and rating of perceived exertion (RPE) during exercise were compared between the trials. RESULTS: Energy expenditure (EE) increased with speed in each trial. However, the Walk trial had a significantly higher EE than the Run trial at speeds exceeding 92 ± 2 % of the maximal walking speed (MWS, p < 0.01). Similarly, carbohydrate (CHO) oxidation was significantly higher in the Walk trial than in the Run trial at above 92 ± 2 %MWS in males (p < 0.001) and above 93 ± 1 %MWS in females (p < 0.05). CONCLUSION: These findings suggest that EE and CHO oxidation during walking increase non-linearly with speed, and walking at a fast speed causes greater metabolic responses than running at the equivalent speed in young participants.

Augmented muscle deoxygenation during repeated sprint exercise with post-exercise blood flow restriction.

Blood flow restriction (BFR) during low-intensity exercise has been known to be a potent procedure to alter metabolic and oxygen environments in working muscles. Moreover, the use of BFR during inter-set rest periods of repeated sprint exercise has been recently suggested to be a potent procedure for improving training adaptations. The present study was designed to determine the effect of repeated sprint exercise with post-exercise BFR (BFR during rest periods between sprints) on muscle oxygenation in working muscles. Eleven healthy males performed two different conditions on different days: either repeated sprint exercise with BFR during rest periods between sets (BFR condition) or without BFR (CON condition). A repeated sprint exercise consisted of three sets of 3 × 6-s maximal sprints (pedaling) with 24s rest periods between sprints and 5 min rest periods between sets. In BFR condition, two min of BFR (100-120 mmHg) for both legs was conducted between sets. During the exercise, power output and arterial oxygen saturation (SpO2 ) were evaluated. Muscle oxygenation for the vastus lateralis muscle, exercise-induced changes in muscle blood flow, and muscle oxygen consumption were measured. During BFR between sets, BFR condition presented significantly higher deoxygenated hemoglobin + myoglobin (p < 0.01) and lower tissue saturation index (p < 0.01) than those in CON condition. However, exercise-induced blood lactate elevation and reduction of blood pH did not differ significantly between the conditions. Furthermore, power output throughout nine sprints did not differ significantly between the two conditions. In conclusion, repeated sprint exercise with post-exercise BFR augmented muscle deoxygenation and local hypoxia, without interfering power output.

Two-step screening method to identify α-synuclein aggregation inhibitors for Parkinson's disease.

Parkinson's disease is a neurodegenerative disease characterized by the formation of neuronal inclusions of α-synuclein in patient brains. As the disease progresses, toxic α-synuclein aggregates transmit throughout the nervous system. No effective disease-modifying therapy has been established, and preventing α-synuclein aggregation is thought to be one of the most promising approaches to ameliorate the disease. In this study, we performed a two-step screening using the thioflavin T assay and a cell-based assay to identify α-synuclein aggregation inhibitors. The first screening, thioflavin T assay, allowed the identification of 30 molecules, among a total of 1262 FDA-approved small compounds, which showed inhibitory effects on α-synuclein fibrilization. In the second screening, a cell-based aggregation assay, seven out of these 30 candidates were found to prevent α-synuclein aggregation without causing substantial toxicity. Of the seven final candidates, tannic acid was the most promising compound. The robustness of our screening method was validated by a primary neuronal cell model and a Caenorhabditis elegans model, which demonstrated the effect of tannic acid against α-synuclein aggregation. In conclusion, our two-step screening system is a powerful method for the identification of α-synuclein aggregation inhibitors, and tannic acid is a promising candidate as a disease-modifying drug for Parkinson's disease.

Muscle Oxygenation during Repeated Cycling Sprints in a Combined Hot and Hypoxic Condition.

The aim of the present study was to examine the effects of a combined hot and hypoxic environment on muscle oxygenation and performance during repeated cycling sprints. In a single-blind, counterbalanced, cross-over research design, 10 male athletes performed three sets of 3 × 10-s maximal pedaling interspersed with 40-s recovery between sprints under four different environments. Each condition consisted of a control (CON; 20°C, 20.9% FiO2), normobaric hypoxia (HYP; 20°C, 14.5% FiO2), hot (HOT; 35°C, 20.9% FiO2), and combined hot and normobaric hypoxia (HH; 35°C, 14.5% FiO2). Power output and vastus lateralis muscle oxygenation were measured. Peak power output was significantly higher in HOT (892±27 W) and HH (887±24 W) than in CON (866±25 W) and HYP (859±25 W) during the first set (p<0.05). The increase in total hemoglobin during recovery periods was larger in HH than in HYP (p<0.05), while change in tissue saturation index was smaller in HYP than in CON and HOT (p<0.05). The findings suggest that the combination of hot and hypoxia during repeated cycling sprints presented different characteristics for muscle metabolism and power output compared to temperature or altitude stressor alone.

Ground reaction force and electromyograms of lower limb muscles during fast walking.

Background: Physically active status is an important contributor to individual health. Walking is regarded as commonly accepted exercise for exercise promotion. Particularly, interval fast walking (FW), consisting of alternating between fast and slow walking speeds, has gained popularity from practical viewpoints. Although previous studies have determined the short- and long-term effects of FW programs on endurance capacity and cardiovascular variables, factors affecting these outcomes have not been clarified. In addition to physiological variables, understanding of mechanical variables and muscle activity during FW would be a help to understand characteristics of FW. In the present study, we compared the ground reaction force (GRF) and lower limb muscle activity between fast walking (FW) and running at equivalent speeds. Method: Eight healthy men performed slow walking (45% of the maximum walking speed; SW, 3.9 ± 0.2 km/h), FW (85% of the maximum walking speed, 7.4 ± 0.4 km/h), and running at equivalent speeds (Run) for 4 min each. GRF and average muscle activity (aEMG) were evaluated during the contact, braking, and propulsive phases. Muscle activities were determined for seven lower limb muscles: gluteus maximus (GM), biceps femoris (BF), rectus femoris (RF), vastus lateralis (VL), gastrocnemius medialis (MG), soleus (SOL), and tibialis anterior (TA). Results: The anteroposterior GRF was greater in FW than in Run during the propulsive phase (p < 0.001), whereas the impact load (peak and average vertical GRF) was lower in FW than in Run (p < 0.001). In the braking phase, lower leg muscle aEMGs were higher during Run than during SW and FW (p < 0.001). However, in the propulsive phase, soleus muscle activity was greater during FW than during Run (p < 0.001). aEMG of tibialis anterior was higher during FW than during SW and Run in the contact phase (p < 0.001). No significant difference between FW and Run was observed for HR and RPE. Conclusion: These results suggest that the average muscle activities of lower limbs (e.g., gluteus maximus, rectus femoris, and soleus) during the contact phase were comparable between FW and running, however, the activity patterns of lower limb muscles differed between FW and running, even at equivalent speeds. During running, muscles were mainly activated in the braking phase related to impact. In contrast, during FW, soleus muscle activity during the propulsive phase was increased. Although cardiopulmonary response was not different between FW and running, exercise using FW might be useful for health promotion among individuals who cannot exercise at high-intensity.

Development of HANABI, an ultrasonication-forced amyloid fibril inducer.

Amyloid fibrils involved in amyloidoses are crystal-like aggregates, which are formed by breaking supersaturation of denatured proteins. Ultrasonication is an efficient method of agitation for breaking supersaturation and thus inducing amyloid fibrils. By combining an ultrasonicator and a microplate reader, we developed the HANABI (HANdai Amyloid Burst Inducer) system that enables high-throughput analysis of amyloid fibril formation. Among high-throughput approaches of amyloid fibril assays, the HANABI system has advantages in accelerating and detecting spontaneous amyloid fibril formation. HANABI is also powerful for amplifying a tiny amount of preformed amyloid fibrils by seeding. Thus, HANABI will contribute to creating therapeutic strategies against amyloidoses by identifying their biomarkers.