Quantitative orbital tightening for pain assessment using machine learning with DeepLabCut.

Pain assessment in animal models is essential for understanding mechanisms underlying pathological pain and developing effective pain medicine. The grimace scale (GS), facial expression features in pain such as orbital tightening (OT), is a valuable measure for assessing pain in animal models. However, the classical grimace scale for pain assessment is labor-intensive, subject to subjectivity and inconsistency, and is not a quantitative measure. In the present study, we utilized machine learning with DeepLabCut to annotate the superior and inferior eyelid margins and the medial and lateral canthus of the eyes in animals' video images. Based on the annotation, we quantified the eyelid distance and palpebral fissure width of the animals' eyes so that the degree of OT in animals with pain could be measured and described quantitatively. We established criteria for the inclusion and exclusion of the annotated images for quantifying OT, and validated our quantitative grimace scale (qGS) in the mice with pain caused by capsaicin injections in the orofacial or hindpaw regions, the Nav1.8-ChR2 mice following orofacial noxious stimulation with laser light, and the oxaliplatin-treated mice following tactile stimulation with a von Frey filament. We showed that both the eyelid distance and the palpebral fissure width were shortened significantly in the animals in pain compared to the control animals without nociceptive stimulation. Collectively, the present study has established a quantitative orbital tightening for pain assessment in mice using DeepLabCut, providing a new tool for pain assessment in preclinical studies with mice.

Development of a noninvasive olfactory stimulation fMRI system in marmosets.

Olfactory dysfunction is associated with aging and the earliest stages of neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases; it is thought to be an early biomarker of cognitive decline. In marmosets, a small non-human primate model used in brain research, olfactory pathway activity during olfactory stimulation has not been well studied because of the difficulty in clearly switching olfactory stimuli inside a narrow MRI. Here, we developed an olfactory-stimulated fMRI system using a small-aperture MRI machine. The olfactory presentation system consisted of two tubes, one for supply and one for suction of olfactory stimulants and a balloon valve. A balloon valve installed in the air supply tube controlled the presentation of the olfactory stimulant, which enabled sharp olfactory stimulation within MRI, such as 30 s of stimulation repeated five times at five-minute intervals. The olfactory stimulation system was validated in vivo and in a simulated system. fMRI analysis showed a rapid increase in signal values within 30 s of olfactory stimulation in eight regions related to the sense of smell. As these regions include those associated with Alzheimer's and Parkinson's diseases, olfactory stimulation fMRI may be useful in clarifying the relationship between olfactory dysfunction and dementia in non-human primates.

Early resolution of bowel urgency by budesonide foam enema results in improved quality of life in patients with ulcerative colitis: a multicenter prospective observational study.

Background/Aims: Bowel urgency is an important symptom for quality of life determination in patients with ulcerative colitis (UC). Few clinical studies have focused on bowel urgency as an efficacy endpoint. Budesonide foam enema has shown efficacy for clinical and endoscopic improvement in mild-to-moderate UC. We evaluated the improvement of clinical symptoms (bowel urgency), safety, and treatment impact of twice-daily budesonide foam enema on the quality of life in patients with UC. Methods: This open-label, multicenter, prospective observational study comprised a 4-week observation period assessing the effectiveness and safety of twice-daily budesonide foam enema. Mild-to-moderate UC patients who had bowel urgency were included. Patients collected data daily in an electronic patient-reported outcome system or logbooks. The primary endpoint was the rate of resolution of bowel urgency at the end of the 4-week observation period. The rate of bowel incontinence was also assessed. Results: Sixty-one patients were enrolled. Of patients with a final evaluation, the rate of resolution of bowel urgency was 58.5% (31/53; 95% confidence interval, 44.1%-71.9%). Bowel urgency decreased over time, with a significant difference observed on day 7 versus day 0. Bowel incontinence showed a decreasing trend from day 5, with a significant difference confirmed on day 12 versus day 0. The clinical remission rate was 64.4% (38/59; 95% confidence interval, 50.9%-76.4%). One adverse event not related to budesonide rectal foam occurred. Conclusions: The findings suggest that bowel urgency can be improved early with twice-daily budesonide foam enema. No new safety signals were observed.

Distinct local and global functions of mouse Aß low-threshold mechanoreceptors in mechanical nociception.

The roles of Aß low-threshold mechanoreceptors (LTMRs) in transmitting mechanical hyperalgesia and in alleviating chronic pain have been of great interest but remain contentious. Here we utilized intersectional genetic tools, optogenetics, and high-speed imaging to specifically examine functions of SplitCre labeled mouse Aß-LTMRs in this regard. Genetic ablation of SplitCre-Aß-LTMRs increased mechanical nociception but not thermosensation in both acute and chronic inflammatory pain conditions, indicating a modality-specific role in gating mechanical nociception. Local optogenetic activation of SplitCre-Aß-LTMRs triggered nociception after tissue inflammation, whereas their broad activation at the dorsal column still alleviated mechanical hypersensitivity of chronic inflammation. Taking all data into consideration, we propose a model, in which Aß-LTMRs play distinctive local and global roles in transmitting or alleviating mechanical hyperalgesia of chronic pain, respectively. Our model suggests a strategy of global activation plus local inhibition of Aß-LTMRs for treating mechanical hyperalgesia.

Clinical Pharmacology of the Antibody-Drug Conjugate Enfortumab Vedotin in Advanced Urothelial Carcinoma and Other Malignant Solid Tumors.

Enfortumab vedotin is an antibody-drug conjugate comprised of a human monoclonal antibody directed to Nectin-4 and monomethyl auristatin E (MMAE), a microtubule-disrupting agent. The objectives of this review are to summarize the clinical pharmacology of enfortumab vedotin monotherapy and demonstrate that the appropriate dose has been selected for clinical use. Pharmacokinetics (PK) of enfortumab vedotin (antibody-drug conjugate and total antibody) and free MMAE were evaluated in five clinical trials of patients with locally advanced or metastatic urothelial carcinoma (n = 748). Intravenous enfortumab vedotin 0.5-1.25 mg/kg on days 1, 8, and 15 of a 28-day cycle showed linear, dose-proportional PK. No significant differences in exposure or safety of enfortumab vedotin and free MMAE were observed in mild, moderate, or severe renal impairment versus normal renal function. Patients with mildly impaired versus normal hepatic function had a 37% increase in area under the concentration-time curve (0-28 days), a 31% increase in maximum concentration of free MMAE, and a similar adverse event profile. No clinically significant PK differences were observed based on race/ethnicity with weight-based dosing, and no clinically meaningful QT prolongation was observed. Concomitant use with dual P-glycoprotein and strong cytochrome P450 3A4 inhibitors may increase MMAE exposure and the risk of adverse events. Approximately 3% of patients developed antitherapeutic antibodies against enfortumab vedotin 1.25 mg/kg. These findings support enfortumab vedotin 1.25 mg/kg monotherapy on days 1, 8, and 15 of a 28-day cycle. No dose adjustments are required for patients with renal impairment or mild hepatic impairment, or by race/ethnicity.

Structural Aspects of Ambient-Temperature Densification of Highly Crack-Resistant Borosilicate and Aluminoborosilicate Glasses: Two Case Studies Examined by Solid-State NMR.

The structural aspects of ambient-temperature densification via pressurization at 25 GPa were studied by solid-state NMR for two case studies: An alkaline earth boroaluminosilicate glass with the composition 6CaO-3SrO-1BaO-10Al2O3-10B2O3-70SiO2 (labeled SAB) and a sodium magnesium borosilicate glass with the composition 10Na2O-10MgO-20B2O3-60SiO2 (labeled MNBS). For SAB glass, cold pressurization results in significant increases in the average coordination numbers of both boron and aluminum, in line with previous results found in hot-compressed alkali aluminoborosilicate glasses. In addition, 27Al/11B dipolar recoupling experiments reveal a significant decrease in the 11B/27Al dipolar interaction strength upon pressurization, suggesting that the higher-coordinated boron and aluminum species experience weaker magnetic interactions. While this is an expected consequence of the longer internuclear distances involving higher coordination states, the magnitude of the effect also is consistent with a decrease of average B-O-Al internuclear connectivity. By conjecture, a decreased B-O-Al connectivity may present a mechanism of plastic flow inhibiting crack initiation in aluminoborosilicate glasses. In the case of the MNBS glass, no change in the average boron coordination number was observed within experimental error; however, densification increases the extent of B-O-Si connectivity at the expense of small ring structures with dominant B-O-B connectivity. With regard to boron coordination, the data obtained for both case studies differ from those previously found in a series of alkali borosilicate glasses, which had shown an unexpected decrease in N4 upon increased pressure. The results of the present study highlight the importance of changes of medium-range order regarding densification.

Effects of inflammation on the properties of Nav1.8-ChR2-positive and Nav1.8-ChR2-negative afferent mechanoreceptors in the hindpaw glabrous skin of mice.

We recently used Nav1.8-ChR2 mice in which Nav1.8-expressing afferents were optogenetically tagged to classify mechanosensitive afferents into Nav1.8-ChR2-positive and Nav1.8-ChR2-negative mechanoreceptors. We found that the former were mainly high threshold mechanoreceptors (HTMRs), while the latter were low threshold mechanoreceptors (LTMRs). In the present study, we further investigated whether the properties of these mechanoreceptors were altered following tissue inflammation. Nav1.8-ChR2 mice received a subcutaneous injection of saline or Complete Freund's Adjuvant (CFA) in the hindpaws. Using the hind paw glabrous skin-tibial nerve preparation and the pressure-clamped single-fiber recordings, we found that CFA-induced hind paw inflammation lowered the mechanical threshold of many Nav1.8-ChR2-positive Aß-fiber mechanoreceptors but heightened the mechanical threshold of many Nav1.8-ChR2-negative Aß-fiber mechanoreceptors. Spontaneous action potential impulses were not observed in Nav1.8-ChR2-positive Aß-fiber mechanoreceptors but occurred in Nav1.8-ChR2-negative Aß-fiber mechanoreceptors with a lower mechanical threshold in the saline goup, and a higher mechanical threshold in the CFA group. No significant change was observed in the mechanical sensitivity of Nav1.8-ChR2-positive and Nav1.8-ChR2-negative Aδ-fiber mechanoreceptors and Nav1.8-ChR2-positive C-fiber mechanoreceptors following hind paw inflammation. Collectively, inflammation significantly altered the functional properties of both Nav1.8-ChR2-positive and Nav1.8-ChR2-negative Aß-fiber mechanoreceptors, which may contribute to mechanical allodynia during inflammation.

ASICs mediate fast excitatory synaptic transmission for tactile discrimination.

Tactile discrimination, the ability to differentiate objects' physical properties such as texture, shape, and edges, is essential for environmental exploration, social interaction, and early childhood development. This ability heavily relies on Merkel cell-neurite complexes (MNCs), the tactile end-organs enriched in the fingertips of humans and the whisker hair follicles of non-primate mammals. Although recent studies have advanced our knowledge on mechanical transduction in MNCs, it remains unknown how tactile signals are encoded at MNCs. Here, using rodent whisker hair follicles, we show that tactile signals are encoded at MNCs as fast excitatory synaptic transmission. This synaptic transmission is mediated by acid-sensing ion channels (ASICs) located on the neurites of MNCs, with protons as the principal transmitters. Pharmacological inhibition or genetic deletion of ASICs diminishes the tactile encoding at MNCs and impairs tactile discrimination in animals. Together, ASICs are required for tactile encoding at MNCs to enable tactile discrimination in mammals.

Prediction and prevention system for Severe Acute Respiratory Syndrome CoronaVirus 2 infection by preempting the onset of a cough.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is fast becoming one of the most significant infections worldwide. Of all the causes of SARS-CoV-2 infection, airborne-droplet infection via coughing is the most common. Therefore, if predicting the onset of a cough and preventing infection were possible, it would have a globally positive impact. Here, we describe a new prediction and prevention system for SARS-CoV-2 infection. Usually, air is inhaled prior to coughing, and the cough, which contains droplets of the virus, then occurs during acute exhalation. Therefore, if we can predict the onset of a cough, we can prevent the spread of SARS-CoV-2. At Tohoku University, a diagnosis system for evaluating swallowing motions and peripheral circulation has already been developed, and our prediction system can be integrated into this system. Using three-dimensional human body imaging, we developed a prediction system for preempting the onset of a cough. If we can predict the onset a cough, we can prevent the spread of SARS-CoV-2 infection, by decreasing the shower of virally active airborne droplets. Here, we describe the newly developed prediction and prevention system for SARS-CoV-2 infection that preempts the onset of a cough.Clinical Relevance- If predicting the onset of a cough and preventing infection were possible, it would have a globally positive impact. Here, we describe the newly developed prediction and prevention system for SARS-CoV-2 infection.

Oncological outcomes of concurrent chemoradiotherapy with docetaxel, cisplatin, and 5-fluorouracil for locally advanced squamous cell carcinoma of the external auditory canal: A single-center study.

BACKGROUND: Squamous cell carcinoma of the external auditory canal (EACSCC) is a rare condition. However, a standard treatment has not yet been established. We retrospectively evaluated the efficacy, adverse events, and feasibility of TPF-CCRT (concomitant chemoradiotherapy with docetaxel, cisplatin, and 5-fluorouracil) in patients with advanced EACSCC. METHODS: Thirty-five consecutive patients with advanced EACSCC (T3, T4) initially treated with TPF-CCRT at Kobe University Hospital were included. T4 diseases with invasion of the brain, internal carotid artery, or internal jugular vein were classified as T4b, and those without these features were classified as T4a. RESULTS: Five-year overall survival rates for T3 and T4 were 100% and 64.2%, respectively. A significant difference was observed between T4a and T4b (82.4% vs. 30%, p = 0.007). Five-year progression-free survival rates of T3, T4a, and T4b were 100%, 68%, and 20% (p = 0.022), respectively. CONCLUSIONS: TPF-CCRT should be considered as a plausible treatment option for advanced EACSCC.

Distinct Local and Global Functions of Aß Low-Threshold Mechanoreceptors in Mechanical Pain Transmission.

The roles of Aß low-threshold mechanoreceptors (LTMRs) in transmitting mechanical hyperalgesia and in alleviating chronic pain have been of great interest but remain contentious. Here we utilized intersectional genetic tools, optogenetics, and high-speed imaging to specifically examine functions of SplitCre labeled Aß-LTMRs in this regard. Genetic ablation of SplitCre-Aß-LTMRs increased mechanical pain but not thermosensation in both acute and chronic inflammatory pain conditions, indicating their modality-specific role in gating mechanical pain transmission. Local optogenetic activation of SplitCre-Aß-LTMRs triggered nociception after tissue inflammation, whereas their broad activation at the dorsal column still alleviated mechanical hypersensitivity of chronic inflammation. Taking all data into consideration, we propose a new model, in which Aß-LTMRs play distinctive local and global roles in transmitting and alleviating mechanical hyperalgesia of chronic pain, respectively. Our model suggests a new strategy of global activation plus local inhibition of Aß-LTMRs for treating mechanical hyperalgesia.

Distinct Local and Global Functions of Aß Low-Threshold Mechanoreceptors in Mechanical Pain Transmission.

The roles of Aß low-threshold mechanoreceptors (LTMRs) in transmitting mechanical hyperalgesia and in alleviating chronic pain have been of great interest but remain contentious. Here we utilized intersectional genetic tools, optogenetics, and high-speed imaging to specifically examine functions of Split Cre labeled Aß-LTMRs in this regard. Genetic ablation of Split Cre -Aß-LTMRs increased mechanical pain but not thermosensation in both acute and chronic inflammatory pain conditions, indicating their modality-specific role in gating mechanical pain transmission. Local optogenetic activation of Split Cre -Aß-LTMRs triggered nociception after tissue inflammation, whereas their broad activation at the dorsal column still alleviated mechanical hypersensitivity of chronic inflammation. Taking all data into consideration, we propose a new model, in which Aß-LTMRs play distinctive local and global roles in transmitting and alleviating mechanical hyperalgesia of chronic pain, respectively. Our model suggests a new strategy of global activation plus local inhibition of Aß-LTMRs for treating mechanical hyperalgesia.

Fibcare® shows correlation with fibrinogen levels by the Clauss method during cardiopulmonary bypass.

Central laboratory measurements are time consuming, while rapid fibrinogen level measurements within the operating room improve transfusion strategies. We aimed to clarify the correlation between fibrinogen concentrations (measured using Fibcare® and the Clauss fibrinogen assay in a central laboratory) during cardiovascular surgery with cardiopulmonary bypass. Data of patients whose Fibcare, traditional laboratory-based testing, and thromboelastographic results were measured using the same blood sample during cardiopulmonary bypass from February 2021 to January 2022 were retrospectively examined. We analyzed correlation in categories of body temperature during cardiopulmonary bypass: total cases, mild hypothermia (28-34°C), and moderate or severe hypothermia (<28°C). The Clauss fibrinogen assay was performed in 123 cases, Fibcare in 107, and thromboelastography in 91. For mild hypothermia, moderate or severe hypothermia, and overall, the root mean squared error and R-square in Fibcare were 16.1 and 0.86, 13.1 and 0.87, and 14.9 and 0.87, respectively, and for thromboelastography, they were 3.26 and 0.74, 2.70 and 0.79, and 3.08 and 0.75, respectively. A significant relationship was noted between Fibcare and Claus fibrinogen analysis regardless of body temperature during cardiopulmonary bypass. The measurement of fibrinogen levels using Fibcare allows for faster transfusion preparation than that of the traditional Clauss fibrinogen assay.

Densification of Sodium Borosilicate Glasses at Ambient Temperature: Structural Investigations by Solid-State Nuclear Magnetic Resonance and Raman Scattering.

Alkali-borosilicate glasses with composition (80-x)SiO2-xB2O3-20Na2O (10 ≤ x ≤ 30) were subjected to a 25 GPa compression and decompression at room temperature, resulting in density increases between 1.4% and 1.9%. The structural changes associated with this process have been investigated and compared with uncompressed glasses having the same thermal history. Systematic trends are identified, using Raman scattering and multinuclear solid-state Nuclear Magnetic Resonance (ssNMR). Perhaps counterintuitively, pressurization tends to increase the concentration of three-coordinated boron species (B(III) units) at the expense of four-coordinated boron (B(IV) units). 23Na NMR spectra show a systematic shift toward higher frequencies in the pressurized glasses, consistent with shorter average Na-O distances. The results are consistently explained in terms of a breakage of Si-O-B4 linkages resulting in the formation of nonbridging oxygen species. Pressure effects on the spectra are reversed by annealing the glasses at their respective glass transition temperatures.

Properties of Nav1.8ChR2-positive and Nav1.8ChR2-negative afferent mechanoreceptors in the hindpaw glabrous skin of mice.

Nav1.8-positive afferent fibers are mostly nociceptors playing a role in mediating thermal and mechanical pain, but mechanoreceptors within these afferents have not been fully investigated. In this study, we generated mice expressing channel rhodopsin 2 (ChR2) in Nav1.8-positive afferents (Nav1.8ChR2), which showed avoidance responses to mechanical stimulation and nocifensive responses to blue light stimulation applied to hindpaws. Using ex vivo hindpaw skin-tibial nerve preparations made from these mice, we characterized properties of mechanoreceptors on Nav1.8ChR2-positive and Nav1.8ChR2-negative afferent fibers that innervate the hindpaw glabrous skin. Of all Aß-fiber mechanoreceptors, small portion was Nav1.8ChR2-positive. Of all Aδ-fiber mechanoreceptors, more than half was Nav1.8ChR2-positive. Of all C-fiber mechanoreceptors, almost all were Nav1.8ChR2-positive. Most Nav1.8ChR2-positive Aß-, Aδ-, and C-fiber mechanoreceptors displayed slowly adapting (SA) impulses in response to sustained mechanical stimulation, and their mechanical thresholds were high in the range of high threshold mechanoreceptors (HTMRs). In contrast, sustained mechanical stimulation applied to Nav1.8ChR2-negative Aß- and Aδ-fiber mechanoreceptors evoked both SA and rapidly adapting (RA) impulses, and their mechanical thresholds were in the range of low threshold mechanoreceptors (LTMRs). Our results provide direct evidence that in the mouse glabrous skin, most Nav1.8ChR2-negative Aß-, Aδ-fiber mechanoreceptors are LTMRs involving in the sense of touch, whereas Nav1.8ChR2-positive Aß-, Aδ-, and C-fiber mechanoreceptors are mainly HTMRs involving in mechanical pain.

Diagnosis and Clinical Features of Perianal Lesions in Newly Diagnosed Crohn's Disease: Subgroup Analysis from Inception Cohort Registry Study of Patients with Crohn's Disease (iCREST-CD).

BACKGROUND AND AIMS: Perianal lesion is a refractory phenotype of Crohn's disease [CD] with significantly diminished quality of life. We evaluated the clinical characteristics of perianal lesions in newly diagnosed CD patients and the impact of perianal lesions on the quality of life in Japanese patients with CD. METHODS: Patients newly diagnosed with CD after June 2016 were included between December 2018 and June 2020 from the Inception Cohort Registry Study of Patients with CD [iCREST-CD]. RESULTS: Perianal lesions were present in 324 [48.2%] of 672 patients with newly diagnosed CD; 71.9% [233/324] were male. The prevalence of perianal lesions was higher in patients aged <40 years vs ≥40 years, and it decreased with age. Perianal fistula [59.9%] and abscess [30.6%] were the most common perianal lesions. In multivariate analyses, male sex, age <40 years and ileocolonic disease location were significantly associated with a high prevalence of perianal lesions, whereas stricturing behaviour and alcohol intake were associated with low prevalence. Fatigue was more frequent [33.3% vs 21.6%] while work productivity and activity impairment-work time missed [36.3% vs 29.5%] and activity impairment [51.9% vs 41.1%] were numerically higher in patients with than those without perianal lesions. CONCLUSIONS: At the time of CD diagnosis, approximately half of the patients had perianal lesions; perianal abscesses and perianal fistulas were the most common. Young age, male sex, disease location and behaviour were significantly associated with the presence of perianal lesions. The presence of perianal lesion was associated with fatigue and impairment of daily activities. CLINICAL TRIALS REGISTRY: University Hospital Medical Information Network Clinical Trials Registry System [UMIN-CTR, UMIN000032237].

Angular Tuning Properties of Low Threshold Mechanoreceptors in Isolated Rat Whisker Hair Follicles.

Angular tuning is preferential sensory response to a directional stimulus and is observed in the whisker tactile system. In whisker hair follicles, there are at least three types of low threshold mechanoreceptors (LTMRs): rapidly adapting (RA), slowly adapting type 1 (SA1), and slowly adapting type 2 (SA2). These LTMRs display angular tuning but their properties remain incompletely studied. Here, we used isolated rat whisker hair follicles and pressure-clamped single-fiber recordings to study angular tuning of these LTMRs. Angular tuning was determined with impulses elicited by ramp-and-hold deflection of whisker hair in 24 directions each at 15° for a total of 360°. We show that RA display impulses during ramp-up, both ramp-up and ramp-down, or ramp-down dynamic phases. Both SA1 and SA2 respond to angular stimuli with slowly adapting impulses in most angles. However, SA1 and SA2 show rapidly adapting responses in other angles. All the three types of LTMRs display strong angular tuning, and there is no significant difference in angular tuning index among them. Population wise, the majority of SA1 are tuned in the caudal direction, a large part of SA2 is tuned in the rostral direction, and RAs are tuned in multiple directions. In the angles showing strong tuning, the three LTMRs respond to increased stimulation amplitudes with increased impulse numbers in a hyperbola relationship, and the responsiveness based on impulse numbers is SA2 > SA1 > RA. Our findings provide new information on angular tuning properties of LTMRs in whisker hair follicles and help to understand directional encoding.

In Vitro Modelling for Bulging Sinus Effects of an Expanded Polytetrafluoroethylene Valved Conduit Based on High-Speed 3D Leaflet Evaluation.

The study aimed to develop a pulmonary circulatory system capable of high-speed 3D reconstruction of valve leaflets to elucidate the local hemodynamic characteristics in the valved conduits with bulging sinuses. Then a simultaneous measurement system for leaflet structure and pressure and flow characteristics was designed to obtain valve leaflet dynamic behaviour with different conduit structures. An image preprocessing method was established to obtain the three leaflets behaviour simultaneously for one sequence with two leaflets images from each pair of three high-speed cameras. Firstly, the multi-digital image correlation analyses were performed, and then the valve leaflet structure was measured under the static condition with fixed opening angles in the water-filled visualization chamber and the pulsatile flow tests simulating paediatric pulmonary flow conditions in the different types of conduit structures; with or without bulging sinuses. The results showed the maximum 3D reconstruction error to be around 0.06 mm. In the steady flow test, the evaluation of opening angles under the different flow rates conditions was achieved. In the pulsatile flow test, each leaflet's opening and closing behaviours were successfully reconstructed simultaneously at the high-frequency recording rate of 960fps. Therefore, the system developed in this study confirms the design evaluation method of an ePTFE valved conduit behaviour with leaflet structures interacting with local fluid dynamics in the vicinity of valves. Clinical Relevance- The system reveals the bulging sinus effects on ePTFE valve leaflet motion by the 3D reconstruction using multi-camera high-speed sequential imaging in vitro.