Discovery of benzimidazole derivatives as orally active renin inhibitors: Optimization of 3,5-disubstituted piperidine to improve pharmacokinetic profile.

We previously identified 2-tert-butyl-4-[(3-methoxypropyl)amino]-N-(2-methylpropyl)-N-[(3S,5R)-5-(morpholin-4-ylcarbonyl)piperidin-3-yl]pyrimidine-5-carboxamide 3 as a potent renin inhibitor. Since 3 showed unacceptably low bioavailability (BA) in rats, structural modification, using SBDD and focused on physicochemical properties was conducted to improve its PK profile while maintaining renin inhibitory activity. Conversion of the amino group attached at the 4-position of pyrimidine to methylene group improved PK profile and decreased renin inhibitory activity. New central cores with carbon side chains were explored to improve potency. We had designed a series of 5-membered azoles and fused heterocycles that interacted with the lipophilic S3 pocket. In the course of modification, renin inhibitory activity was enhanced by the formation of an additional hydrogen bonding with the hydroxyl group of Thr77. Consequently, a series of novel benzimidazole derivatives were discovered as potent and orally bioavailable renin inhibitors. Among those, compound 13 exhibited more than five-fold of plasma renin inhibition than aliskiren in cynomolgus monkeys at dose ratio.

Effects of fearful face presentation time and observer's eye movement on the gaze cue effect.

BACKGROUND: There are many conflicting findings on the gaze cueing effect (GCE) of emotional facial expressions. This study aimed to investigate whether an averted gaze, accompanied by a fearful expression of different durations, could enhance attentional orientation, as measured by a participant's eye movements. METHODS: Twelve participants (3 females) completed the gaze cue task, reacting to a target location after observing changes in the gaze and expression of a face illustrated on a computer screen. Meanwhile, participants' eye movements were monitored by electrooculography. The GCE was calculated by reaction time as an indicator of attention shift. RESULTS: The analysis of the overall data did not find a significant effect of fearful facial expressions on the GCE. However, analysis of trial data that excluded a participant's eye movement data showed that brief (0, 100 ms) presentation of the fearful facial expression enhanced the GCE compared to that during a neutral facial expression, although when the presentation time of the fearful expression was increased to 200 or 400 ms, the GCE of the fearful expression was at the same level as when model showed a neutral expression. CONCLUSIONS: The results suggest that the attention-enhancing effect of gaze cues induced by rapidly presented fearful expressions occurs only when the effect of eye movement trials is excluded. This effect may be mediated by reflexively neural circuits in the amygdala that process threatening stimuli. However, as the expression duration increased, the fearful expression's attention-enhancing effect decreased. We suggest that future studies on the emotion modulation of GCE should consider the negative effects of participants' saccades and blinks on the experimental results.

Prostaglandin E2 promotes wound-induced migration of intestinal subepithelial myofibroblasts via EP2, EP3, and EP4 prostanoid receptor activation.

Intestinal subepithelial myofibroblasts (ISMFs) are mesenchymal cells that reside in the subepithelial region throughout the intestine. When the intestine is damaged, the migratory and mitotic responses of ISMFs are crucial for wound closure. However, their mechanism of action remains unknown. We have investigated the role of cyclooxygenase (COX) and its metabolite prostaglandin E(2) (PGE(2)) in the wound repair process of bovine ISMFs. The action of a mechanical scratch in a layer of ISMFs in cell culture elevated the levels of both COX-2 mRNA expression and PGE(2) secretion 1 and 6 h after the event. After 24 h ISMFs had migrated to and reduced the wounded area around the site of the scratch. Treatment with the COX-1/2 inhibitor indomethacin, the COX-2 inhibitor 3-(4-methylsulphonylphenyl)-4-phenyl-5-trifluoromethylisoxazole (CAY10404), or E prostanoid receptor 2 to 4 (EP2-EP4) antagonists significantly inhibited wound repair. Conversely, inhibition of wound closure by indomethicin was reversed by treatment with PGE(2) or agonists of the receptors EP2, EP3, or EP4 but not of EP1. Although EP2 to EP4 stimulation did not influence ISMF proliferation, it did stimulate ISMF migration in the transwell cell migration assay. It is noteworthy that cell migration stimulated by EP2 and EP4 was inhibited by the tyrosine kinase receptor inhibitor genistein and also by (Z)-3-[2,4-dimethyl-5-(2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl]-propionic acid (SU6668). However, cell migration stimulated by EP3 was unaffected. Reverse transcription-polymerase chain reaction showed EP2 or EP4 stimulation elevated the level of mRNA expression for fibroblast growth factor-2, which stimulates ISMF migration. Collectively, COX-2-dependent PGE(2) secretion promotes wound healing by ISMFs. PGE(2)-EP3 signaling may directly stimulate ISMF migration. PGE(2)-EP2/4 signaling indirectly stimulates ISMF migration by elevating the level of growth factor secretion.

Isolation and characterization of bovine intestinal subepithelial myofibroblasts.

Intestinal subepithelial myofibroblasts (ISMFs) are mesenchymal cells that exist under the epithelium of intestines. Primarily isolated ISMFs from rodents have been applied to experiments. However, due to the size of their intestines, the available cell number is limited. Thus, we attempted to isolate ISMFs from bovine colon as an alternative material. After detachment of smooth muscle and epithelial layers, colonic mucosa was explanted. After 2-week incubation, alpha-SMA+ / vimentin+ / desmin(-) ISMFs were harvested and applied for experiments. First we examined the effect of cell passage on morphology and proliferation activity of bovine ISMFs. Although 3rd and 7th passage bovine ISMFs did not exhibit any changes, 11th passage ISMFs showed rounded enlarged shape and lost proliferation potential. On the contrary, rat ISMFs displayed the above senescent changes at earlier passage (passage 4). In intracellular Ca2+ concentration measurement, bioactive substances (0.3-1 microM ATP, 0.1-1 microM serotonin, 10-100 nM endothelin-1, and 1-10 nM bradykinin) dose-dependently induced an increase in intracellular Ca2+ concentration in bovine ISMFs (passage 3 and 7). However, at passage 11, impairment in intracellular Ca2+ responses was observed. Thus, bovine ISMFs might be a novel useful tool with long life span and good cellular responses to investigate physiological/pathophysiological roles of ISMFs.

What is the significance of the traditional pinching mode of holding chopsticks?

BACKGROUND: The purpose of this study was to clarify the influence of manipulation mode of chopsticks on the learning process, using assessment of task performance and electromyography, and to understand the significance of the traditional manipulation mode from the viewpoint of physiological anthropology. Previous studies have described two modes of manipulating chopsticks, the traditional pincers-pinching mode and the scissors-pinching mode. METHODS: We conducted experiments with two conditions of holding chopsticks: scissors mode and pincers mode. Eight subjects participated and were assigned to these modes, and they learned handling tasks in their assigned mode for 5 days with the non-dominant hand. We measured task execution times and conducted electromyography of the following muscles: first dorsalis interosseus, flexor pollicis brevis, flexor digiti minimi brevis, flexor digitorum superficialis, and extensor digitorum. RESULTS: The training effects were found in each mode. The pincers mode showed significantly shorter task performance times than did scissors mode. On electromyography, significant increases in activity of flexor digiti minimi brevis and tended an increase in flexor digitorum superficialis and a decrease in extensor digitorum occurred in pincers mode but not in scissors mode. CONCLUSIONS: The traditional mode of holding chopsticks was associated with not only high task performance but also an advantage in terms of learning motor control.

Quizartinib, a selective FLT3 inhibitor, maintains antileukemic activity in preclinical models of RAS-mediated midostaurin-resistant acute myeloid leukemia cells.

FLT3 internal tandem duplication (ITD) mutations are associated with poor prognosis in patients with acute myeloid leukemia (AML). In this preclinical study, we characterized the binding affinity and selectivity of quizartinib, a small-molecule inhibitor of FLT3, and AC886, the active metabolite of quizartinib, compared with those of other FLT3 inhibitors. Selectivity profiling against >400 kinases showed that quizartinib and AC886 were highly selective against FLT3. Quizartinib and AC886 inhibited FLT3 signaling pathways in FLT3-ITD-mutated AML cells, leading to potent growth inhibition with IC50 values of <1 nM. When quizartinib was administered to mice bearing FLT3-ITD mutated tumors, AC886 was rapidly detected and tumor regression was observed at doses of ≥1 mg/kg without severe body weight loss. In addition, quizartinib inhibited the viability of midostaurin-resistant MOLM-14 cells and exerted potent antitumor activity in mouse xenograft models without severe body weight loss, while midostaurin and gilteritinib did not show significant antitumor effects. This is the first detailed characterization of quizartinib and AC886 in comparison with other FLT3 inhibitors under the same experimental conditions. Preclinical antileukemic activity in midostaurin-resistant FLT3-ITD-mutated AML cells suggests the potential value of quizartinib following midostaurin failure in patients with FLT3-ITD mutated AML.

RUVBL1/RUVBL2 ATPase Activity Drives PAQosome Maturation, DNA Replication and Radioresistance in Lung Cancer.

RUVBL1 and RUVBL2 (collectively RUVBL1/2) are essential AAA+ ATPases that function as co-chaperones and have been implicated in cancer. Here we investigated the molecular and phenotypic role of RUVBL1/2 ATPase activity in non-small cell lung cancer (NSCLC). We find that RUVBL1/2 are overexpressed in NSCLC patient tumors, with high expression associated with poor survival. Utilizing a specific inhibitor of RUVBL1/2 ATPase activity, we show that RUVBL1/2 ATPase activity is necessary for the maturation or dissociation of the PAQosome, a large RUVBL1/2-dependent multiprotein complex. We also show that RUVBL1/2 have roles in DNA replication, as inhibition of its ATPase activity can cause S-phase arrest, which culminates in cancer cell death via replication catastrophe. While in vivo pharmacological inhibition of RUVBL1/2 results in modest antitumor activity, it synergizes with radiation in NSCLC, but not normal cells, an attractive property for future preclinical development.

Cathodal transcranial direct-current stimulation over right posterior parietal cortex enhances human temporal discrimination ability.

BACKGROUND: Time perception associated with durations from 1 s to several minutes involves activity in the right posterior parietal cortex (rPPC). It is unclear whether altering the activity of the rPPC affects an individual's timing performance. Here, we investigated the human timing performance under the application of transcranial direct-current stimulation (tDCS) that altered the neural activities of the rPPC. METHODS: We measured the participants' duration-discrimination threshold by administering a behavioral task during the tDCS application. The tDCS conditions consisted of anodal, cathodal, and sham conditions. The electrodes were placed over the P4 position (10-20 system) and on the left supraorbital forehead. On each task trial, the participant observed two visual stimuli and indicated which was longer. The amount of difference between the two stimulus durations was varied repeatedly throughout the trials according to the participant's responses. The correct answer rate of the trials was calculated for each amount of difference, and the minimum amount with the correct answer rate exceeding 75% was selected as the threshold. The data were analyzed by a linear mixed-effects models procedure. RESULTS: Nineteen volunteers participated in the experiment. We excluded three participants from the analysis: two who reported extreme sleepiness while performing the task and one who could recognize the sham condition correctly with confidence. Our analysis of the 16 participants' data showed that the average value of the thresholds observed under the cathodal condition was lower than that of the sham condition. This suggests that inhibition of the rPPC leads to an improvement in temporal discrimination performance, resulting in improved timing performance. CONCLUSIONS: In the present study, we found a new effect that cathodal tDCS over the rPPC enhances temporal discrimination performance. In terms of the existence of anodal/cathodal tDCS effects on human timing performance, the results were consistent with a previous study that investigated temporal reproduction performance during tDCS application. However, the results of the current study further indicated that cathodal tDCS over the rPPC increases accuracy of observed time duration rather than inducing an overestimation as a previous study reported.

Additive Effects of Sinusoidal Lower Body Negative Pressure on Cardiovascular Responses.

BACKGROUND: Sinusoidal lower body negative pressure (SLBNP) has been used to investigate the cardiovascular response to slow periodic changes in blood shifts, but measurements of slow fluctuations take a long time if measured for each period of SLBNP separately. Our study aimed to investigate whether the cardiovascular responses to superimposed SLBNP (S-SLBNP), which is expected to reduce the measurement time, are different from responses measured individually. METHODS: S-SLBNP was configured by superimposing two conventional SLBNPs (C-SLBNP) at 180-s and 30-s periods in the pressure range from 0 to -25 mmHg. As the S-SLBNP has double the static load of C-SLBNP, we also used offset SLBNP (O-SLBNP), which has the same static load level as S-SLBNP. Heart rate (HR), thoracic impedance (Z0), and mean arterial pressure (MAP) were measured from 11 male subjects. The transfer functions of gains from MAP to HR (Gain-HR/MAP) and from Z0 to HR (Gain-HR/Z0) were calculated as indexes of arterial baroreflex and cardiopulmonary baroreflex regulation of HR, respectively. RESULTS: The Gain-HR/MAP in the 180-s period (2.11 ± 0.17 bpm/mmHg; mean ± SEM) was larger than that of the 30-s period (1.04 ± 0.09 bpm/mmHg); however, there was no significant difference between the SLBNP conditions. The Gain-HR/Z0 in C-SLBNP (9.37 ± 1.47 bpm/ohm) was smaller than that of the other conditions [18.46 ± 2.45 bpm/ohm (O-SLBNP); 16.09 ± 2.29 bpm/ohm (S-SLBNP)]. DISCUSSION: Using S-SLBNP could reduce the measurement time needed to examine the arterial baroreflex. However, the cardiopulmonary baroreflex was modified by the static load of SLBNP.Ishibashi K, Oyama F, Yoshida H, Iwanaga K. Additive effects of sinusoidal lower body negative pressure on cardiovascular responses. Aerosp Med Hum Perform. 2017; 88(2):137-141.

Mast cell-derived prostaglandin D2 inhibits colitis and colitis-associated colon cancer in mice.

Compared with prostaglandin E2, which has an established role in cancer, the role of the COX metabolite prostaglandin D2 (PGD2) in chronic inflammation leading to tumorigenesis is uncertain. In this study, we investigated the role of PGD2 in colitis and colitis-associated colon cancer (CAC) using genetically modified mice and an established model of inflammatory colon carcinogenesis. Systemic genetic deficiency in hematopoietic PGD synthase (H-PGDS) aggravated colitis and accelerated tumor formation in a manner associated with increased TNFα expression. Treatment with a TNFα receptor antagonist attenuated colitis regardless of genotype. Histologic analysis revealed that infiltrated mast cells strongly expressed H-PGDS in inflamed colons. Mast cell-specific H-PGDS deficiency also aggravated colitis and accelerated CAC. In contrast, treatment with a PGD2 receptor agonist inhibited colitis and CAC. Together, our results identified mast cell-derived PGD2 as an inhibitor of colitis and CAC, with implications for its potential use in preventing or treating colon cancer.

Purinergic P2Y1 receptor signaling mediates wound stimuli-induced cyclooxygenase-2 expression in intestinal subepithelial myofibroblasts.

Intestinal subepithelial myofibroblasts (ISMFs) are crucial for barrier formation against inflammatory stimuli. Physical injury induces cyclooxygenase-2 (COX-2) expression, which accelerates wound healing by ISMFs. However, the mechanism of COX-2 induction remains unclear. Physically damaged cells release ATP. Here, we investigate the role of ATP-purinergic signaling in wound-induced COX-2 induction in ISMFs. By 24h post-injury, bovine ISMFs had migrated to and closed the wounded area. A COX inhibitor, indomethacin or a purinergic P2 receptor antagonist, suramin, inhibited wound healing. However, additional treatment with indomethacin did not influence wound healing in suramin-treated ISMFs. RT-PCR showed an increase in COX-2 mRNA expression 2h post-injury, which was inhibited by suramin. These results suggest that ATP mediates wound-induced COX-2 elevation. We next assessed the contribution of various purinergic receptors in COX-2 induction. An ATP analog, ATPγS and a purinergic P2Y1, 11-13 receptors agonist, ADP, were among the agents tested which increased COX-2 expression. ATPγS-induced COX-2 mRNA expression was suppressed by suramin or a purinergic P2Xs, P2Y1, 4, 6, and 13 receptors antagonist, PPADS. These data suggest the involvement of Gq-coupled purinergic P2Y1 receptor or Gi-coupled purinergic P2Y13 receptor in COX-2 induction. U73122, an inhibitor of phospholipase C, which is a downstream signal of Gq protein, showed suppression of COX-2 mRNA expression. However, pertussis toxin, a Gi inhibitor, did not show suppression. We also revealed that inhibitors of p38 MAPK and PKC inhibited ATPγS-induced COX-2 mRNA expression. Collectively, purinergic P2Y1 receptor signaling mediates wound-induced COX-2 expression through p38 MAPK and PKC pathways in ISMFs.

Differences in cardiovascular and central nervous system responses to periods of mental work with a break.

The purpose of the present study was to examine how an inserted break influences the cardiovascular and central nervous system responses during periods of mental work. Twelve males conducted two 20-min periods of mental work with a 3-min break between them. Cardiovascular and central nervous system responses were measured continuously. In comparison to the baseline, cardiovascular responses increased continuously even after the inserted break, while, on the contrary, central nervous system activity did not significantly increase during the work periods but relaxed during the break. The work performance increased during the second work period. These results suggest that the inserted break proposed by VDT guidelines in Japan was effective in relaxing the central nervous system but was insufficient to prevent the increase in cardiovascular load. The results also imply that taking rests frequently is important not only to maintaining performance but also to preventing cumulative physiological workloads.

Interaction between musical emotion and facial expression as measured by event-related potentials.

We examined the integrative process between emotional facial expressions and musical excerpts by using an affective priming paradigm. Happy or sad musical stimuli were presented after happy or sad facial images during electroencephalography (EEG) recordings. We asked participants to judge the affective congruency of the presented face-music pairs. The congruency of emotionally congruent pairs was judged more rapidly than that of incongruent pairs. In addition, the EEG data showed that incongruent musical targets elicited a larger N400 component than congruent pairs. Furthermore, these effects occurred in nonmusicians as well as musicians. In sum, emotional integrative processing of face-music pairs was facilitated in congruent music targets and inhibited in incongruent music targets; this process was not significantly modulated by individual musical experience. This is the first study on musical stimuli primed by facial expressions to demonstrate that the N400 component reflects the affective priming effect.

Comparison of cardiovascular response to sinusoidal and constant lower body negative pressure with reference to very mild whole-body heating.

BACKGROUND: The purpose of the present study was to compare sinusoidal versus constant lower body negative pressure (LBNP) with reference to very mild whole-body heating. Sinusoidal LBNP has a periodic load component (PLC) and a constant load component (CLC) of orthostatic stress, whereas constant LBNP has only a CLC. We tested two sinusoidal patterns (30-s and 180-s periods with 25 mmHg amplitude) of LBNP and a constant LBNP with -25 mmHg in 12 adult male subjects. RESULTS: Although the CLC of all three LBNP conditions were configured with -25 mmHg, the mean arterial pressure (MAP) results showed a significantly large decrease from baseline in the 30-s period condition (P <0.01). In contrast, the other cardiovascular indices (heart rate (HR), stroke volume (SV), cardiac output (CO), basal thoracic impedance (Z(0)), total peripheral resistance (TPR), the natural logarithmic of the HF component (lnHF), and LF/HF (ln(LF/HF))) of heart rate variability (HRV) showed relatively small variations from baseline in the 30-s period condition (P <0.01). The result of the gain and phase of transfer function at the sinusoidal period of LBNP showed that the very mild whole-body heating augmented the orthostatic responses. CONCLUSION: These results revealed that the effect of the CLC of LBNP on cardiovascular adjustability was attenuated by the addition of the PLC to LBNP. Based on the results of suppressed HRV response from baseline in the 30-s period condition, we suggest that the attenuation may be caused by the suppression of the vagal responsiveness to LBNP.

Circulatory and central nervous system responses to different types of mental stress.

The purpose of the present study was to compare the physiological responses to different types of mental stress encountered in the workplace. Circulatory and central nervous system responses were examined in 8 healthy males by exposing them to 20-min of white noise (80 dB(A)) and 20-min of computer-based mental arithmetic tasks as models of vascular and cardiac stress, respectively. The results indicated that both cardiac and vascular stresses increased blood pressure and showed a cumulative effect as exposure period was extended. Heart rate and prefrontal oxygenated hemoglobin levels (measured by NIRS) increased in the face of cardiac stress but were not clearly altered by vascular stress and indicated that cardiac stress higher cardiac response and requires more oxygen supply to the brain. As the central nervous system responded, an event-related potential P300 component was elicited by an auditory oddball task presented before and after each stress. The P300 amplitude increased for both stresses. However, P300 latency increased in response to cardiac stress but decreased with vascular stress in the left prefrontal. Thus, the circulatory and central nervous system responses to cardiac stress and to vascular stress may have different underlying mechanisms, and measuring physiological indices appears to be an effective method by which to evaluate the influence of mental stress.

ATP induces contraction mediated by the P2Y(2) receptor in rat intestinal subepithelial myofibroblasts.

Intestinal subepithelial myofibroblasts (IMFs) exist just under the epithelial membrane directly facing the mucosal microvascular capillary surface distributed in the lamina propria. In the gastrointestinal tract, ATP is released from epithelial and endothelial cells in response to mechanical stimuli. Although it has been reported that mechanical stimuli evoke synchronized Ca(2+) waves in cultured IMFs, the contractile responses by ATP stimulation have not been examined. The aim of this study was to clarify the mechanism of the contraction of IMFs in response to ATP. ATP (1-30µM) induced contraction in a concentration-dependent manner. These contractions were inhibited by LaCl(3) (100-300µM) and by Ca(2+)-free solution (0.5mM EGTA). Fura-2/Ca(2+) signals indicated that ATP (1-10µM) elicited transient increases in intracellular Ca(2+) concentration ([Ca(2+)](i)). In addition, αß-methylene-ATP (10, 30 and 300µM), a broad spectrum P2X agonist at a concentration higher than 100µM, induced neither contraction nor [Ca(2+)](i) rise. UTP (1-30µM), a selective P2Y(2) and P2Y(4) agonist in rodent, induced concentration-dependent contractions and [Ca(2+)](i) increases, whereas ADP and UDP (10µM) did not induce contractions. Pretreatment with suramin (30-100µM), a relatively selective P2Y(2) antagonist, strongly inhibited ATP- and UTP-induced contractions and [Ca(2+)](i) increases. However, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS: 10-30µM), a receptor antagonist for several P2X and P2Y but less effective to P2Y(2) receptor, failed to inhibit ATP- and UTP-induced contractions and [Ca(2+)](i) increases. By RT-PCR, mRNA expressions of the P2Y(1) and P2Y(2) receptors, but not P2Y(4) or P2Y(6), were detected in IMFs. These results suggest that ATP induces [Ca(2+)](i)-dependent contraction in IMFs, which is mediated through the P2Y(2) receptor.

The reproducibility of cardiovascular response to a mental task.

The present study investigated the reproducibility of cardiovascular responses by presenting a mental task to eight healthy subjects four times at intervals of one year and of several days. Subjects performed a mental subtraction task at the same time of day and under the same conditions. For each experiment day, systolic and diastolic blood pressure (SBP and DBP), mean arterial pressure (MAP), cardiac output (CO), heart rate (HR), stroke volume (SV), and total peripheral resistance (TPR) were measured for a 5-minute baseline period, a 5-minute task period, and a 10-minute recovery period. To examine the reproducibility of cardiovascular response, two-way factorial ANOVA (subject factor and experiment day factor) was conducted, and the intra-class correlation coefficient (ICC) was calculated. The results showed that there were no significant differences between experiment days separated by one year or between those separated by several days for all indexes. The ICCs were high for blood pressures (SBP, DBP and MAP, ICCs>0.8) and cardiac responses (HR, SV, and CO, ICCs>0.7) for both the interval of one year and that of several days. For the total peripheral resistance, ICC was only high at the interval of several days (>0.7). In conclusion, the reproducibility of the blood pressure response to a mental task was proven, and in case of the changed blood pressure, the cardiac responses were also reproduced for intervals of one year and of several days.

Relationship between muscular strength and deflection characteristics of the center of foot pressure during landing after crossover stepping in the elderly.

This study aimed to investigate the relationship between the muscular strength of the lower extremity in a load side and the characteristics of center of foot pressure (COP) during landing after crossover stepping in the elderly. The study population comprised 8 elderly subjects (average age, 75.8+/-8.0 years) and 9 young individuals (average age, 21.6+/-2.5 years). Using a separation-type force plate, we measured the deflection characteristics of the COP; these were defined by the root mean square of positional change (COP-RMS) and the deflection velocity of the COP (COP-Vel) during landing after crossover stepping. Furthermore, we measured the muscular strength of the lower extremity by using a hand-held dynamometer. By using multiple regression analysis, we detected the calculated muscular strength as the independent variable of the deflection characteristics of the COP. Compared to the young group the elderly group showed significantly higher anterior-posterior COP-RMS values (p<0.05) and lower lateral COP-Vel values (p<0.001). In the elderly, the muscular strengths of the tibialis anterior and adductor magnus were detected as a significant independent variable of the anterior-posterior COP-RMS (R(2)=0.85, R(2)=0.76, p<0.01) and lateral COP-Vel (R(2)=0.75, R(2)=0.65, p<0.05), respectively. With regard to the COP deflection characteristics during landing after crossover stepping in the elderly, we recognized the diagnostic character of the anterior-posterior COP-RMS and lateral COP-Vel. Furthermore, it was suggested that the muscular strengths of the tibialis anterior and adductor magnus played a role in regulating the COP deflection characteristics.

Effects of strap support in a hand-held device on the muscular activity in female workers assessed by electromyography and subjective rating.

The present study evaluates the potential mitigation of physical workload when using strap support for a portable device. The experiments were designed as consecutive sessions over a 2-h period. Electromyogram signals were recorded from four muscles of six subjects. The perceived level of fatigue on the whole body as well as in the shoulder, arm, lower back and legs was assessed using Borg's CR-10 scale. All subjects were tested under eight experimental conditions. Results indicated that the biceps brachii muscle displayed significantly lower activity with strap support than without a strap. In the experiments with and without a strap, different levels of force were imposed on the various muscles, which caused changes in the distribution of the physical load. Although the role of the strap might seem evident, using strap support did not always decrease the sensation of fatigue. However, for short-term tasks, using a strap may be recommended.

Carbachol induces Ca(2+)-dependent contraction via muscarinic M2 and M3 receptors in rat intestinal subepithelial myofibroblasts.

Intestinal myofibroblasts (IMFs) that exist adjacent to the basement membrane of intestines have contractility and contribute to physical barriers of the intestine. Nerve endings distribute adjacent to IMFs, suggesting neurotransmitters may influence IMFs motility; however, there is no direct evidence showing the interaction. Here, we isolated IMFs from rat colon and investigated the effect of acetylcholine on IMFs contractility. In the collagen gel contraction assay, carbachol (1 - 10 microM) and the muscarinic receptor agonist bethanechol (30 - 300 microM) dose-dependently induced IMFs contraction. Pretreatment with the muscarinic receptor antagonist atropine (1 - 10 nM) inhibited carbachol-induced contraction. In RT-PCR, mRNA expression of all muscarinic receptor subtypes (M(1) - M(5)) was detected in IMFs. Subsequently we found pretreatment with the muscarinic M(2) receptor antagonist 11-([2-[(diethylamino)methyl]-1-piperdinyl]acetyl)-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepine-6-one (AF-DX116) (10 and 30 nM) or the muscarinic M(3) receptor antagonist 4-diphenylacetoxy-N-methyl-piperidine (4-DAMP) (3 and 10 nM) dose-dependently inhibited carbachol-induced contraction. In Ca(2+) measurement, 1 - 10 microM carbachol and 30 - 300 microM bethanechol elevated the intracellular Ca(2+) concentration ([Ca(2+)](i)) in IMFs. Atropine (10 nM) eliminated carbachol-induced [Ca(2+)](i) elevation. The Ca(2+)-channel blocker LaCl(3) (3 microM) abolished carbachol-induced [Ca(2+)](i) elevation and contraction. Furthermore, AF-DX116 and 4-DAMP dose-dependently inhibited the carbachol-induced [Ca(2+)](i) elevation. These observations suggest that acetylcholine elicits Ca(2+)-dependent IMF contraction through muscarinic M(2) and M(3) receptors.