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Squatting, a traditional resistance exercise classified as strength training, relies on anaerobic pathways, but its aerobic aspects remain unclear. We examined heart rate and oxygen demand during squats, exploring variations across different strength statuses. It fills gaps in understanding the cardiorespiratory effects of squatting, especially during multiple sets. Twenty-two young healthy resistance trained men (age: 28 ± 4 years) participated. Maximal oxygen consumption (VÌO2max) and 1 repetition maximum (RM) of squat were measured. Participants performed 5 sets of squat exercises at 65% of 1RM for 10 repetitions with 3-min rest intervals. Heart rate and pulmonary gas exchange were measured during the squat exercise. Participants were divided into high strength (HS; upper 50%) and low strength (LS; lower 50%) groups based on a median split of their 1 RM squat values (normalized to their body weight). During 5 sets of squat exercise, oxygen consumption (VÌO2) increased up to 47.8 ± 8.9 ml/kg/min, corresponding to 100.6% of predetermined VÌO2max. The HS group achieved a greater highest point of VÌO2 in relation to VÌO2max than the LS group (108.0 vs. 93.7%). During the exercise intervals, VÌO2 exceeded VÌCO2, while during the rest intervals, VÌCO2 surpassed VÌO2. Our findings suggest that the oxygen demand during squatting is notably substantial, which may vary according to the training status.
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Frequência Cardíaca , Consumo de Oxigênio , Treinamento Resistido , Humanos , Masculino , Consumo de Oxigênio/fisiologia , Adulto , Frequência Cardíaca/fisiologia , Exercício Físico/fisiologia , Adulto Jovem , Troca Gasosa Pulmonar/fisiologiaRESUMO
PURPOSE: Results of initial endoscopic biopsy of gastric lesions often differ from those of the final pathological diagnosis. We evaluated whether an artificial intelligence-based gastric lesion detection and diagnostic system, ENdoscopy as AI-powered Device Computer Aided Diagnosis for Gastroscopy (ENAD CAD-G), could reduce this discrepancy. MATERIALS AND METHODS: We retrospectively collected 24,948 endoscopic images of early gastric cancers (EGCs), dysplasia, and benign lesions from 9,892 patients who underwent esophagogastroduodenoscopy between 2011 and 2021. The diagnostic performance of ENAD CAD-G was evaluated using the following real-world datasets: patients referred from community clinics with initial biopsy results of atypia (n=154), participants who underwent endoscopic resection for neoplasms (Internal video set, n=140), and participants who underwent endoscopy for screening or suspicion of gastric neoplasm referred from community clinics (External video set, n=296). RESULTS: ENAD CAD-G classified the referred gastric lesions of atypia into EGC (accuracy, 82.47%; 95% confidence interval [CI], 76.46%-88.47%), dysplasia (88.31%; 83.24%-93.39%), and benign lesions (83.12%; 77.20%-89.03%). In the Internal video set, ENAD CAD-G identified dysplasia and EGC with diagnostic accuracies of 88.57% (95% CI, 83.30%-93.84%) and 91.43% (86.79%-96.07%), respectively, compared with an accuracy of 60.71% (52.62%-68.80%) for the initial biopsy results (P<0.001). In the External video set, ENAD CAD-G classified EGC, dysplasia, and benign lesions with diagnostic accuracies of 87.50% (83.73%-91.27%), 90.54% (87.21%-93.87%), and 88.85% (85.27%-92.44%), respectively. CONCLUSIONS: ENAD CAD-G is superior to initial biopsy for the detection and diagnosis of gastric lesions that require endoscopic resection. ENAD CAD-G can assist community endoscopists in identifying gastric lesions that require endoscopic resection.
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Inteligência Artificial , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/patologia , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/cirurgia , Estudos Retrospectivos , Feminino , Masculino , Gastroscopia/métodos , Pessoa de Meia-Idade , Idoso , Diagnóstico por Computador/métodos , Biópsia/métodos , Lesões Pré-Cancerosas/patologia , Lesões Pré-Cancerosas/diagnóstico , Lesões Pré-Cancerosas/cirurgia , Endoscopia do Sistema Digestório/métodos , Detecção Precoce de Câncer/métodosRESUMO
The countermovement jump is a popular measurement modality to evaluate muscle power in sports and exercise. Muscle power is essential to achieve a high jump, yet the well-coordinated movement of the body segments, which optimizes the stretch-shortening cycle (SSC) effects, is also required. Among the proposed explanations of SSC effects, this study investigated whether the ankle joint kinematics, kinetics, and muscle-tendon interaction depend on the level of jump skill and the jump task. Sixteen healthy males were grouped as a function of their jump height (High jumpers; greater than 50 cm, Low jumpers; less than 50 cm). They were instructed to jump with two intensities; light effort (20 % of their height) and maximal effort. Joint kinematics and kinetics of the lower limbs were analyzed using a 3-dimensional motion analysis system. The muscle-tendon interaction was investigated using B-mode real-time ultrasonography. As the jump intensity increased, all participants jumped with increased joint velocity and power. However, the high jumper shows less fascicle shortening velocity (-0.2 ± 0.1 m/s) than the low jumper group (-0.3 ± 0.1 m/s) and greater tendon velocity, which indicated the capability of elastic energy recoil. In addition, the delayed onset time of ankle extension in the high jumper implies better use of the catapult mechanism. The findings of this study showed that the muscle-tendon interaction differs depending on the jump skill level, suggesting a more efficient neuromuscular control in skilled jumpers.
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Músculo Esquelético , Esportes , Masculino , Humanos , Músculo Esquelético/diagnóstico por imagem , Músculo Esquelético/fisiologia , Tendões/diagnóstico por imagem , Tendões/fisiologia , Articulação do Tornozelo/fisiologia , Exercício Físico/fisiologiaRESUMO
Fluorescence probe is one of the most powerful tools for cellular imaging. Here, three phospholipid-mimicking fluorescent probes (FP1-FP3) comprising fluorescein and two lipophilic groups of saturated and/or unsaturated C18 fatty acids were synthesized, and their optical properties were investigated. Like in biological phospholipids, the fluorescein group acts as a hydrophilic polar headgroup and the lipid groups act as hydrophobic non-polar tail groups. Laser confocal microscope images illustrated that FP3, which contains both saturated and unsaturated lipid tails, showed great uptake into the canine adipose-derived mesenchymal stem cells.
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BACKGROUND AND AIM: Although resting heart rate (RHR) is associated with prevalence and incidence of diabetes, whether it is associated with undiagnosed diabetes is still unclear. We aimed to investigate whether the RHR is associated with the prevalence of undiagnosed diabetes in a large Korean national dataset. METHODS AND RESULTS: The Korean National Health and Nutrition Examination Survey data from 2008 to 2018 were used. After screening, 51,637 participants were included in this study. The odds ratios and 95% confidence intervals (CIs) for undiagnosed diabetes were calculated using multivariable-adjusted logistic regression analyses. Analyses showed that participants with a RHR of ≥90 bpm showed a 4.00- (95% CI: 2.77-5.77) and 3.21-times (95% CI: 2.01-5.14) higher prevalence of undiagnosed diabetes for men and women, respectively, than those with a RHR of <60 bpm. The linear dose-response analyses showed that each 10-bpm increment in RHR was associated with a 1.39- (95% CI: 1.32-1.48) and 1.28-times (95% CI: 1.19-1.37) higher prevalence of undiagnosed diabetes for men and women, respectively. In the stratified analyses, the positive association between RHR and the prevalence of undiagnosed diabetes was tended to be stronger among those who were younger (age: <40 years) and lean (BMI: <23 kg/m2). CONCLUSIONS: Elevated RHR was significantly associated with a higher prevalence of undiagnosed diabetes in Korean men and women, independent of demographic, lifestyle, and medical factors. Accordingly, the value of RHR as a clinical indicator and health marker, especially in reducing the prevalence of undiagnosed diabetes, is suggestible.
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Diabetes Mellitus , Masculino , Humanos , Adulto , Feminino , Prognóstico , Inquéritos Nutricionais , Frequência Cardíaca , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/epidemiologia , República da Coreia/epidemiologia , Fatores de RiscoRESUMO
PURPOSE: To identify important features of lymph node metastasis (LNM) and develop a prediction model for early gastric cancer (EGC) using a gradient boosting machine (GBM) method. MATERIALS AND METHODS: The clinicopathologic data of 2556 patients with EGC who underwent gastrectomy were used as training set and the internal validation set (set 1) at a ratio of 8:2. Additionally, 548 patients with EGC who underwent endoscopic submucosal dissection (ESD) as the initial treatment were included in the external validation set (set 2). The GBM model was constructed, and its performance was compared with that of the Japanese guidelines. RESULTS: LNM was identified in 12.6% (321/2556) of the gastrectomy group (training set & set 1) and 4.3% (24/548) of the ESD group (set 2). In the GBM analysis, the top five features that most affected LNM were lymphovascular invasion, depth, differentiation, size, and location. The accuracy, sensitivity, specificity, and the area under the receiver operating characteristics of set 1 were 0.566, 0.922, 0.516, and 0.867, while those of set 2 were 0.810, 0.958, 0.803, and 0.944, respectively. When the sensitivity of GBM was adjusted to that of Japanese guidelines (beyond the expanded criteria in set 1 [0.922] and eCuraC-2 in set 2 [0.958]), the specificities of GBM in sets 1 and 2 were 0.516 (95% confidence interval, 0.502-0.523) and 0.803 (0.795-0.805), while those of the Japanese guidelines were 0.502 (0.488-0.509) and 0.788 (0.780-0.790), respectively. CONCLUSION: The GBM model showed good performance comparable with the eCura system in predicting LNM risk in EGCs.
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Neoplasias Gástricas , Humanos , Neoplasias Gástricas/cirurgia , Neoplasias Gástricas/patologia , Metástase Linfática , Modelos Logísticos , Fatores de Risco , Excisão de Linfonodo , Estudos RetrospectivosRESUMO
During a vertical drop jump (VDJ), the human neuromuscular system absorbs and reuses external loads applied to the lower extremity by coordinating the musculoskeletal system. This study aims to investigate the influence of the eccentric strength of the knee extensor muscles on the biomechanical factors of a VDJ. Participants were divided into two groups based on the eccentric strength of their knee extension muscles: low eccentric (LECC) and high eccentric (HECC) strength groups. The VDJ joint kinematics and kinetics of the lower extremity, the fascicle behavior of the vastus lateralis, and the muscle activation of the knee extensor muscles were simultaneously recorded during maximum-effort VDJ. Compared with the LECC group, the HECC group showed a higher jump, greater knee and ankle joint stiffness, and smaller fascicle length change. These findings suggest that the eccentric strength capacity of the knee extensor muscles accounts for the different biomechanical strategies (bouncing-type for HECC and absorbing-type for LECC) observed between the groups. Consequently, the eccentric strength of the knee extensor muscle may be an essential factor in determining the biomechanical strategy for VDJ and should be considered in the jumping performance enhancement training paradigm.
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Artropatias , Músculo Esquelético , Fenômenos Biomecânicos , Humanos , Joelho/fisiologia , Articulação do Joelho/fisiologia , Extremidade Inferior , Força Muscular , Músculo Esquelético/fisiologiaRESUMO
This study aimed to investigate the association between domain-specific physical activity (PA) and diabetes in Korean adults. We analyzed 26,653 men and women (aged > 18 years) from the Korea National Health and Nutrition Examination Survey (2014-2018). PA was measured using a validated Global PA Questionnaire. Multivariable logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) after adjustment for various confounders. Transport PA accounted for the majority of total PA (46%, men; 58%, women), followed by leisure-time PA (30%; 22%) and work PA (24%; 20%). In men, ORs (95% CI) of diabetes comparing ≥ 600 metabolic task of equivalent (MET)-min/week vs. no activity were 0.82 (0.71-0.95) for leisure-time PA, 0.85 (0.75-0.96) for transport PA, and 0.88 (0.78-0.99) for leisure-time + transport PA. In women, ORs (95% CI) of diabetes comparing the same groups were 0.73 (0.60-0.89) for leisure-time PA, 0.97 (0.85-1.10) for transport PA, and 0.88 (0.78-1.00) for leisure-time + transport PA. However, work PA showed no association with diabetes. In conclusion, leisure-time PA was inversely associated with diabetes in both men and women, while transport PA was inversely associated only in men. But work PA was not associated with diabetes in Korean adults.
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Povo Asiático , Diabetes Mellitus/epidemiologia , Exercício Físico/fisiologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , República da Coreia/epidemiologiaRESUMO
The purpose of this study was to investigate how the ball position along the mediolateral (M-L) direction of a golfer causes a chain effect in the ground reaction force, body segment and joint angles, and whole-body centre of mass during the golf swing. Twenty professional golfers were asked to complete five straight shots for each 5 different ball positions along M-L: 4.27 cm (ball diameter), 2.14 cm (ball radius), 0 cm (reference position at preferred ball position), - 2.14 cm, and - 4.27 cm, while their ground reaction force and body segment motions were captured. The dependant variables were calculated at 14 swing events from address to impact, and the differences between the ball positions were evaluated using Statistical Parametric Mapping. The left-sided ball positions at address showed a greater weight distribution on the left foot with a more open shoulder angle compared to the reference ball position, whereas the trend was reversed for the right-sided ball positions. These trends disappeared during the backswing and reappeared during the downswing. The whole-body centre of mass was also located towards the target for the left-sided ball positions throughout the golf swing compared to the reference ball position, whereas the trend was reversed for the right-sided ball positions. We have concluded that initial ball position at address can cause a series of chain effects throughout the golf swing.
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An extensive evaluation of low-cost dust sensors was performed using an exponentially decaying particle concentration. A total of 264 sensors including 27 sensors with light-emitting diodes (LEDs) and 237 sensors with laser lighting sources were tested. Those tested sensors were classified into 4 groups based on the deviation from the reference data obtained by a reference instrument. The response linearities of all the tested samples for PM1 , PM2.5 , and PM10 were in excellent agreement with the reference instrument, except a few samples. For the measurements of PM1 and PM2.5 , the lighting source, that is, LED or laser, did not show any significant difference in overall sensor performance. However, LED-based sensors did not perform well for PM10 measurements. The 32, 24, and 16% of all the tested sensors for PM1 , PM2.5 , and PM10 measurement, respectively, are in the category of Class 1 (reference instrument reading ± 20%) requirement. The performance of the low-cost dust sensors for PM10 measurement was relatively less satisfactory.
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Poluição do Ar em Ambientes Fechados/análise , Poeira/análise , Monitoramento Ambiental/métodos , Poluentes Atmosféricos/análise , Monitoramento Ambiental/economia , Monitoramento Ambiental/instrumentação , Material Particulado/análiseRESUMO
The purpose of this study was to investigate address position variables in response to changes in ball position in golfers. Eleven male professional golfers were instructed to perform their golf swing. A three-dimensional motion analysis system, with eight infrared cameras and two force platforms, was used to capture the address positions. A golf ball has a diameter of 4.27 cm, and a radius of 2.14 cm. Even small movements of ball position in the mediolateral (M-L) and anteroposterior (A-P) directions significantly changed the address position. When the ball was moved to the left, the shoulder rotation and club-face aim rotated toward the left of the target, and the left vertical ground reaction force increased. When the ball was moved to the right, the opposite findings were observed. When the ball was moved closer, the trunk, hip, knee, ankle, and absolute arm angle extended; the lie angle of the golf club increased; and the center of pressure moved toward the posterior direction. These changes were reversed when the ball was moved further away. The M-L ball position critically changed the address positions of the upper extremities in the horizontal plane, and the A-P ball position critically changed the angles of whole body parts in the sagittal plane. Furthermore, club-head kinematics at impact such as club-face aim, club path, and angle of attack were significantly changed in the M-L ball position; and club-head speed and angle of attack were significantly changed in the A-P ball position. This in-depth understanding of the address position in association with the ball position could provide valuable data for swing coaches when finding a golfer's optimal address position.
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Golfe/fisiologia , Postura , Equipamentos Esportivos , Adulto , Fenômenos Biomecânicos , Humanos , Perna (Membro) , Masculino , Movimento (Física) , Pressão , Rotação , Ombro , TroncoRESUMO
We aimed to investigate the characteristics of patients with osteoarthritis (OA), using the data of all Koreans registered in the National Health Insurance Sharing Service Database (NHISS DB), and to provide ideal alternative cutoff thresholds for alleviating OA symptoms. Patients with OA (codes M17 and M17.1-M17.9 in the Korean Standard Classification of Disease and Causes of Death) were analyzed using SAS software. Optimal cutoff thresholds were determined using receiver operating characteristic curve analysis. The 50-year age group was the most OA pathogenic group (among 40~70 years, n = 2088). All exercise types affected the change of body mass index (p < 0.05) and the sex difference in blood pressure (BP) (p < 0.01). All types of exercise positively affected the loss of waist circumference and the balance test (standing time on one leg in seconds) (p < 0.01). The cutoff threshold for the time in seconds from standing up from a chair to walking 3 m and returning to the same chair was 8.25 (80% sensitivity and 100% specificity). By using the exercise modalities, categorized multiple variables, and the cutoff threshold, an optimal alternative exercise program can be designed for alleviating OA symptoms in the 50-year age group.
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Big Data , Osteoartrite do Joelho , Curva ROC , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Programas Nacionais de Saúde/estatística & dados numéricos , República da Coreia , Estudos Retrospectivos , Circunferência da CinturaRESUMO
This study aimed to examine whether muscle force and tendon stiffness in a muscle-tendon complex alter synchronously following 8-week whole-body vibration (WBV) training in older people. Forty older women aged 65 years and older were randomly assigned into control (CON, n = 15) and whole-body vibration (WBV) training groups (exposure time, n = 13; vibration intensity, n = 12). For the training groups, a 4-week detraining period was completed following the training period. Throughout the training/detraining period, force of the medial gastrocnemius (MG) muscle and stiffness of the Achilles tendon were assessed four times (0, 4, 8, and 12 weeks) using a combined system of dynamometer and ultrasonography. While muscle force gradually increased throughout the training period (p < .05), a significant increase in tendon stiffness was observed after 8 weeks (p < .05). These findings indicated that, during the early phase of WBV training, muscle force and tendon stiffness changed asynchronously, which might be a factor in possible musculotendinous injuries.
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Tendão do Calcâneo/fisiologia , Força Muscular/fisiologia , Treinamento Resistido/métodos , Vibração/uso terapêutico , Tendão do Calcâneo/diagnóstico por imagem , Idoso , Eletromiografia , Feminino , Humanos , Inquéritos e Questionários , Resultado do TratamentoRESUMO
We propose a new method to estimate joint force using a biomechanical muscle model and peaks of surface electromyography (SEMG). The SEMG measurement was carried out from the first dorsal interosseous muscle during isometric index finger abduction. The SEMG peaks were used as the input of the biomechanical muscle model which is a transfer function to generate the force. The force estimation performance ( R(2) ) was evaluated using the proposed method with nine healthy subjects, and a former method using a mean absolute value (MAV), which is the full-wave rectified and averaged (or low-pass filtered) signal of SEMG in a time window, was compared with the proposed method; the performance of the proposed method (0.94 ± 0.03) was better than that of MAV (0.90 ± 0.02). The proposed method could be widely applied to quantitative analysis of muscle activities based on SEMG.
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Eletromiografia/métodos , Articulações dos Dedos/fisiologia , Dedos/fisiologia , Adulto , Fenômenos Biomecânicos/fisiologia , Feminino , Humanos , Masculino , Adulto JovemRESUMO
We propose a new method - twitch force - for estimation of the muscle behavior during voluntary contraction for assessing localized muscle fatigue. The proposed method uses the sEMG peaks as input and the measured force as output. The twitch force, which is a transfer function to generate force, was estimated during fatiguing contraction. We verified the estimated twitch force based on the measured results with electrical stimulation. The participants performed isometric little finger flexion until exhaustion. SEMG was recorded on the flexor digiti minimi brevis muscle for the proposed method and the electrical stimulation electrodes on the ulnar nerve induced involuntary contraction for reference. As the muscle fatigue level increased, the twitch peaks decreased in both methods. The proposed method can be widely used in the quantitative analysis of muscle fatigue during voluntary contraction.
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Eletromiografia/métodos , Fadiga Muscular/fisiologia , Fenômenos Biomecânicos , Estimulação Elétrica , Eletrodos , Dedos/fisiologia , Humanos , Contração Muscular/fisiologia , Músculo Esquelético/fisiologia , Adulto JovemRESUMO
The objectives of this study was to investigate the acute effects of various magnitudes of tendon strain on the mechanical properties of the human medial gastrocnemius (MG) in vivo during controlled heel-drop exercises. Seven male and seven female volunteers performed two different exercises executed one month apart: one was a heel-drop exercise on a block (HDB), and the other was a heel-drop exercise on level floor (HDL). In each regimen, the subjects completed a session of 150 heel-drop exercises (15 repetitions×10 sets; with a 30 s rest following each set). Before and immediately after the heel-drop exercise, the ankle plantar flexor torque and elongation of the MG were measured using a combined measurement system of dynamometry and ultrasonography and then the MG tendon strain and stiffness were evaluated in each subject. The tendon stiffness measured prior to the exercises was not significantly different between the two groups 23.7±10.6N/mm and 24.1±10.0N/mm for the HDB and HDL, respectively (p>.05). During the heel-drop exercise, it was found that the tendon strain during the heel-drop exercise on a block (8.4±3.7%) was significantly higher than the strain measured on the level floor (5.4±3.8%) (p<.05). In addition, the tendon stiffness following the heel-drop exercise on a block (32.3±12.2N/mm) was significantly greater than the tendon stiffness measured following the heel-drop exercise on the level floor (25.4±11.4N/mm) (p<.05). The results of this study suggest that tendon stiffness immediately following a heel-drop exercise depends on the magnitude of tendon strain.
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Tendão do Calcâneo/fisiologia , Exercício Físico/fisiologia , Calcanhar/fisiologia , Músculo Esquelético/fisiologia , Adaptação Fisiológica , Adulto , Análise de Variância , Tornozelo/fisiologia , Elasticidade , Feminino , Voluntários Saudáveis , Humanos , Masculino , Movimento (Física) , Estresse Mecânico , Torque , Suporte de Carga/fisiologia , Adulto JovemRESUMO
Surface electromygraphy (sEMG) provides information of the neural drive to the muscle, so muscle force estimation by sEMG is of high relevance in biomechanical studies and in bionic applications. Even though mean absolute value (MAV) has been widely used for sEMG amplitude estimation due to the probabilistic nature of sEMG, but it has been used without any comprehensive physiological justification. A physiologically and biomechanically approximate model for the force estimation would enable a clear understanding of the relationships between sEMG and the force, and it can be used as sEMG amplitude estimation method. We proposed a new sEMG amplitude estimation method comprising two procedures: MUAP (motor unit action potential) event detection and muscle force indication using a biomechanical muscle model. The estimation performances were evaluated with nine subjects and compared with MAV. The performance (R(2)) of the proposed method (0.94 ± 0.03) outperformed it of MAV (0.90 ± 0.02). The method we proposed should be widely applicable to quantitatively analysis muscle activities by sEMG.
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Eletromiografia/métodos , Modelos Teóricos , Fenômenos Biomecânicos , Humanos , Músculos/fisiologiaRESUMO
The palmar pinch force estimation is highly relevant not only in biomechanical studies, the analysis of sports activities, and ergonomic design analyses but also in clinical applications such as rehabilitation, in which information about muscle forces influences the physician's decisions on diagnosis and treatment. Force transducers have been used for such purposes, but they are restricted to grasping points and inevitably interfere with the human haptic sense because fingers cannot directly touch the environmental surface. We propose an estimation method of the palmar pinch force using surface electromyography (SEMG). Three myoelectric sites on the skin were selected on the basis of anatomical considerations and a Fisher discriminant analysis (FDA), and SEMG at these sites yields suitable information for pinch force estimation. An artificial neural network (ANN) was implemented to map the SEMG to the force, and its structure was optimized to avoid both under- and over-fitting problems. The resulting network was tested using SEMG signals recorded from the selected myoelectric sites of ten subjects in real time. The training time for each subject was short (approximately 96s), and the estimation results were promising, with a normalized root mean squared error (NRMSE) of 0.081+/-0.023 and a correlation (CORR) of 0.968+/-0.017.
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Algoritmos , Eletromiografia/métodos , Dedos/fisiologia , Modelos Biológicos , Contração Muscular/fisiologia , Músculo Esquelético/fisiologia , Redes Neurais de Computação , Feminino , Humanos , Masculino , Estresse Mecânico , Adulto JovemRESUMO
Atomic force microscopy (AFM) has been used to study the specific interactions between the signal-transducing proteins mammalian phospholipase D1 (PLD1), phospholipase C-gamma1 (PLC-gamma1), and Munc-18-1. To record the forces between them, the Phox homology (PX) domain of PLD1, the Src homology (SH3) domain of PLC-gamma1, and Munc-18-1 were fused with glutathione S-transferase (GST) and immobilized onto reduced glutathione (GSH)-tethered surfaces. In order to enhance the recognition efficiency and avoid undesirable complications, both AFM tips and substrates were first modified with dendrons of two different sizes. Under the employed conditions, the probability of observing an unbinding event increased, most force-distance curves showed the single rupture events, and the unbinding forces were 51 +/- 2 pN for PX-(Munc-18-1) and 42 +/- 2 pN for PX-SH3. To investigate dynamics of these biomolecular interactions, we measured the loading rate dependence of the unbinding forces. The unbinding forces increased linearly with the logarithm of the loading rate, indicating the presence of a single potential barrier in the dissociation energy landscape. The measured off-rate constants (k(off)) at 15 degrees C were 10(-3.4 +/- 0.3) s(-1) for PX-(Munc-18-1) and 10(-1.7 +/- 0.1) s(-1) for PX-SH3. Further, we elucidated the influence of free SH3 and Munc-18-1 on the specific PX-(Munc-18-1) and PX-SH3 interaction, respectively.