Dynamic 3D scanning as a markerless method to calculate multi-segment foot kinematics during stance phase: methodology and first application.

PubMed

Van den Herrewegen, Inge; Cuppens, Kris; Broeckx, Mario; Barisch-Fritz, Bettina; Vander Sloten, Jos; Leardini, Alberto; Peeraer, Louis

2014-08-22

Multi-segmental foot kinematics have been analyzed by means of optical marker-sets or by means of inertial sensors, but never by markerless dynamic 3D scanning (D3DScanning). The use of D3DScans implies a radically different approach for the construction of the multi-segment foot model: the foot anatomy is identified via the surface shape instead of distinct landmark points. We propose a 4-segment foot model consisting of the shank (Sha), calcaneus (Cal), metatarsus (Met) and hallux (Hal). These segments are manually selected on a static scan. To track the segments in the dynamic scan, the segments of the static scan are matched on each frame of the dynamic scan using the iterative closest point (ICP) fitting algorithm. Joint rotations are calculated between Sha-Cal, Cal-Met, and Met-Hal. Due to the lower quality scans at heel strike and toe off, the first and last 10% of the stance phase is excluded. The application of the method to 5 healthy subjects, 6 trials each, shows a good repeatability (intra-subject standard deviations between 1° and 2.5°) for Sha-Cal and Cal-Met joints, and inferior results for the Met-Hal joint (>3°). The repeatability seems to be subject-dependent. For the validation, a qualitative comparison with joint kinematics from a corresponding established marker-based multi-segment foot model is made. This shows very consistent patterns of rotation. The ease of subject preparation and also the effective and easy to interpret visual output, make the present technique very attractive for functional analysis of the foot, enhancing usability in clinical practice. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Comparison between low-cost marker-less and high-end marker-based motion capture systems for the computer-aided assessment of working ergonomics.

PubMed

Patrizi, Alfredo; Pennestrì, Ettore; Valentini, Pier Paolo

2016-01-01

The paper deals with the comparison between a high-end marker-based acquisition system and a low-cost marker-less methodology for the assessment of the human posture during working tasks. The low-cost methodology is based on the use of a single Microsoft Kinect V1 device. The high-end acquisition system is the BTS SMART that requires the use of reflective markers to be placed on the subject's body. Three practical working activities involving object lifting and displacement have been investigated. The operational risk has been evaluated according to the lifting equation proposed by the American National Institute for Occupational Safety and Health. The results of the study show that the risk multipliers computed from the two acquisition methodologies are very close for all the analysed activities. In agreement to this outcome, the marker-less methodology based on the Microsoft Kinect V1 device seems very promising to promote the dissemination of computer-aided assessment of ergonomics while maintaining good accuracy and affordable costs. PRACTITIONER’S SUMMARY: The study is motivated by the increasing interest for on-site working ergonomics assessment. We compared a low-cost marker-less methodology with a high-end marker-based system. We tested them on three different working tasks, assessing the working risk of lifting loads. The two methodologies showed comparable precision in all the investigations.

Hybrid markerless tracking of complex articulated motion in golf swings.

PubMed

Fung, Sim Kwoh; Sundaraj, Kenneth; Ahamed, Nizam Uddin; Kiang, Lam Chee; Nadarajah, Sivadev; Sahayadhas, Arun; Ali, Md Asraf; Islam, Md Anamul; Palaniappan, Rajkumar

2014-04-01

Sports video tracking is a research topic that has attained increasing attention due to its high commercial potential. A number of sports, including tennis, soccer, gymnastics, running, golf, badminton and cricket have been utilised to display the novel ideas in sports motion tracking. The main challenge associated with this research concerns the extraction of a highly complex articulated motion from a video scene. Our research focuses on the development of a markerless human motion tracking system that tracks the major body parts of an athlete straight from a sports broadcast video. We proposed a hybrid tracking method, which consists of a combination of three algorithms (pyramidal Lucas-Kanade optical flow (LK), normalised correlation-based template matching and background subtraction), to track the golfer's head, body, hands, shoulders, knees and feet during a full swing. We then match, track and map the results onto a 2D articulated human stick model to represent the pose of the golfer over time. Our work was tested using two video broadcasts of a golfer, and we obtained satisfactory results. The current outcomes of this research can play an important role in enhancing the performance of a golfer, provide vital information to sports medicine practitioners by providing technically sound guidance on movements and should assist to diminish the risk of golfing injuries. Copyright © 2013 Elsevier Ltd. All rights reserved.

Cloning-independent markerless gene editing in Streptococcus sanguinis: novel insights in type IV pilus biology.

PubMed

Gurung, Ishwori; Berry, Jamie-Lee; Hall, Alexander M J; Pelicic, Vladimir

2017-04-07

Streptococcus sanguinis, a naturally competent opportunistic human pathogen, is a Gram-positive workhorse for genomics. It has recently emerged as a model for the study of type IV pili (Tfp)-exceptionally widespread and important prokaryotic filaments. To enhance genetic manipulation of Streptococcus sanguinis, we have developed a cloning-independent methodology, which uses a counterselectable marker and allows sophisticated markerless gene editing in situ. We illustrate the utility of this methodology by answering several questions regarding Tfp biology by (i) deleting single or mutiple genes, (ii) altering specific bases in genes of interest, and (iii) engineering genes to encode proteins with appended affinity tags. We show that (i) the last six genes in the pil locus harbouring all the genes dedicated to Tfp biology play no role in piliation or Tfp-mediated motility, (ii) two highly conserved Asp residues are crucial for enzymatic activity of the prepilin peptidase PilD and (iii) that pilin subunits with a C-terminally appended hexa-histidine (6His) tag are still assembled into functional Tfp. The methodology for genetic manipulation we describe here should be broadly applicable. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

Cloning-independent markerless gene editing in Streptococcus sanguinis: novel insights in type IV pilus biology

PubMed Central

Gurung, Ishwori; Berry, Jamie-Lee; Hall, Alexander M. J.

2017-01-01

Abstract Streptococcus sanguinis, a naturally competent opportunistic human pathogen, is a Gram-positive workhorse for genomics. It has recently emerged as a model for the study of type IV pili (Tfp)—exceptionally widespread and important prokaryotic filaments. To enhance genetic manipulation of Streptococcus sanguinis, we have developed a cloning-independent methodology, which uses a counterselectable marker and allows sophisticated markerless gene editing in situ. We illustrate the utility of this methodology by answering several questions regarding Tfp biology by (i) deleting single or mutiple genes, (ii) altering specific bases in genes of interest, and (iii) engineering genes to encode proteins with appended affinity tags. We show that (i) the last six genes in the pil locus harbouring all the genes dedicated to Tfp biology play no role in piliation or Tfp-mediated motility, (ii) two highly conserved Asp residues are crucial for enzymatic activity of the prepilin peptidase PilD and (iii) that pilin subunits with a C-terminally appended hexa-histidine (6His) tag are still assembled into functional Tfp. The methodology for genetic manipulation we describe here should be broadly applicable. PMID:27903891

Markerless client-server augmented reality system with natural features

NASA Astrophysics Data System (ADS)

Ning, Shuangning; Sang, Xinzhu; Chen, Duo

2017-10-01

A markerless client-server augmented reality system is presented. In this research, the more extensive and mature virtual reality head-mounted display is adopted to assist the implementation of augmented reality. The viewer is provided an image in front of their eyes with the head-mounted display. The front-facing camera is used to capture video signals into the workstation. The generated virtual scene is merged with the outside world information received from the camera. The integrated video is sent to the helmet display system. The distinguishing feature and novelty is to realize the augmented reality with natural features instead of marker, which address the limitations of the marker, such as only black and white, the inapplicability of different environment conditions, and particularly cannot work when the marker is partially blocked. Further, 3D stereoscopic perception of virtual animation model is achieved. The high-speed and stable socket native communication method is adopted for transmission of the key video stream data, which can reduce the calculation burden of the system.

Nearly automatic motion capture system for tracking octopus arm movements in 3D space.

PubMed

Zelman, Ido; Galun, Meirav; Akselrod-Ballin, Ayelet; Yekutieli, Yoram; Hochner, Binyamin; Flash, Tamar

2009-08-30

Tracking animal movements in 3D space is an essential part of many biomechanical studies. The most popular technique for human motion capture uses markers placed on the skin which are tracked by a dedicated system. However, this technique may be inadequate for tracking animal movements, especially when it is impossible to attach markers to the animal's body either because of its size or shape or because of the environment in which the animal performs its movements. Attaching markers to an animal's body may also alter its behavior. Here we present a nearly automatic markerless motion capture system that overcomes these problems and successfully tracks octopus arm movements in 3D space. The system is based on three successive tracking and processing stages. The first stage uses a recently presented segmentation algorithm to detect the movement in a pair of video sequences recorded by two calibrated cameras. In the second stage, the results of the first stage are processed to produce 2D skeletal representations of the moving arm. Finally, the 2D skeletons are used to reconstruct the octopus arm movement as a sequence of 3D curves varying in time. Motion tracking, segmentation and reconstruction are especially difficult problems in the case of octopus arm movements because of the deformable, non-rigid structure of the octopus arm and the underwater environment in which it moves. Our successful results suggest that the motion-tracking system presented here may be used for tracking other elongated objects.

Markerless human motion tracking using hierarchical multi-swarm cooperative particle swarm optimization.

PubMed

Saini, Sanjay; Zakaria, Nordin; Rambli, Dayang Rohaya Awang; Sulaiman, Suziah

2015-01-01

The high-dimensional search space involved in markerless full-body articulated human motion tracking from multiple-views video sequences has led to a number of solutions based on metaheuristics, the most recent form of which is Particle Swarm Optimization (PSO). However, the classical PSO suffers from premature convergence and it is trapped easily into local optima, significantly affecting the tracking accuracy. To overcome these drawbacks, we have developed a method for the problem based on Hierarchical Multi-Swarm Cooperative Particle Swarm Optimization (H-MCPSO). The tracking problem is formulated as a non-linear 34-dimensional function optimization problem where the fitness function quantifies the difference between the observed image and a projection of the model configuration. Both the silhouette and edge likelihoods are used in the fitness function. Experiments using Brown and HumanEva-II dataset demonstrated that H-MCPSO performance is better than two leading alternative approaches-Annealed Particle Filter (APF) and Hierarchical Particle Swarm Optimization (HPSO). Further, the proposed tracking method is capable of automatic initialization and self-recovery from temporary tracking failures. Comprehensive experimental results are presented to support the claims.

Monitoring tumor motion by real time 2D/3D registration during radiotherapy.

PubMed

Gendrin, Christelle; Furtado, Hugo; Weber, Christoph; Bloch, Christoph; Figl, Michael; Pawiro, Supriyanto Ardjo; Bergmann, Helmar; Stock, Markus; Fichtinger, Gabor; Georg, Dietmar; Birkfellner, Wolfgang

2012-02-01

In this paper, we investigate the possibility to use X-ray based real time 2D/3D registration for non-invasive tumor motion monitoring during radiotherapy. The 2D/3D registration scheme is implemented using general purpose computation on graphics hardware (GPGPU) programming techniques and several algorithmic refinements in the registration process. Validation is conducted off-line using a phantom and five clinical patient data sets. The registration is performed on a region of interest (ROI) centered around the planned target volume (PTV). The phantom motion is measured with an rms error of 2.56 mm. For the patient data sets, a sinusoidal movement that clearly correlates to the breathing cycle is shown. Videos show a good match between X-ray and digitally reconstructed radiographs (DRR) displacement. Mean registration time is 0.5 s. We have demonstrated that real-time organ motion monitoring using image based markerless registration is feasible. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Structure Sensor for mobile markerless augmented reality

NASA Astrophysics Data System (ADS)

Kilgus, T.; Bux, R.; Franz, A. M.; Johnen, W.; Heim, E.; Fangerau, M.; Müller, M.; Yen, K.; Maier-Hein, L.

2016-03-01

3D Visualization of anatomical data is an integral part of diagnostics and treatment in many medical disciplines, such as radiology, surgery and forensic medicine. To enable intuitive interaction with the data, we recently proposed a new concept for on-patient visualization of medical data which involves rendering of subsurface structures on a mobile display that can be moved along the human body. The data fusion is achieved with a range imaging device attached to the display. The range data is used to register static 3D medical imaging data with the patient body based on a surface matching algorithm. However, our previous prototype was based on the Microsoft Kinect camera and thus required a cable connection to acquire color and depth data. The contribution of this paper is two-fold. Firstly, we replace the Kinect with the Structure Sensor - a novel cable-free range imaging device - to improve handling and user experience and show that the resulting accuracy (target registration error: 4.8+/-1.5 mm) is comparable to that achieved with the Kinect. Secondly, a new approach to visualizing complex 3D anatomy based on this device, as well as 3D printed models of anatomical surfaces, is presented. We demonstrate that our concept can be applied to in vivo data and to a 3D printed skull of a forensic case. Our new device is the next step towards clinical integration and shows that the concept cannot only be applied during autopsy but also for presentation of forensic data to laypeople in court or medical education.

SU-G-BRA-05: Application of a Feature-Based Tracking Algorithm to KV X-Ray Fluoroscopic Images Toward Marker-Less Real-Time Tumor Tracking

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nakamura, M; Matsuo, Y; Mukumoto, N

Purpose: To detect target position on kV X-ray fluoroscopic images using a feature-based tracking algorithm, Accelerated-KAZE (AKAZE), for markerless real-time tumor tracking (RTTT). Methods: Twelve lung cancer patients treated with RTTT on the Vero4DRT (Mitsubishi Heavy Industries, Japan, and Brainlab AG, Feldkirchen, Germany) were enrolled in this study. Respiratory tumor movement was greater than 10 mm. Three to five fiducial markers were implanted around the lung tumor transbronchially for each patient. Before beam delivery, external infrared (IR) markers and the fiducial markers were monitored for 20 to 40 s with the IR camera every 16.7 ms and with an orthogonalmore » kV x-ray imaging subsystem every 80 or 160 ms, respectively. Target positions derived from the fiducial markers were determined on the orthogonal kV x-ray images, which were used as the ground truth in this study. Meanwhile, tracking positions were identified by AKAZE. Among a lot of feature points, AKAZE found high-quality feature points through sequential cross-check and distance-check between two consecutive images. Then, these 2D positional data were converted to the 3D positional data by a transformation matrix with a predefined calibration parameter. Root mean square error (RMSE) was calculated to evaluate the difference between 3D tracking and target positions. A total of 393 frames was analyzed. The experiment was conducted on a personal computer with 16 GB RAM, Intel Core i7-2600, 3.4 GHz processor. Results: Reproducibility of the target position during the same respiratory phase was 0.6 +/− 0.6 mm (range, 0.1–3.3 mm). Mean +/− SD of the RMSEs was 0.3 +/− 0.2 mm (range, 0.0–1.0 mm). Median computation time per frame was 179 msec (range, 154–247 msec). Conclusion: AKAZE successfully and quickly detected the target position on kV X-ray fluoroscopic images. Initial results indicate that the differences between 3D tracking and target position would be clinically acceptable.« less

On-patient see-through augmented reality based on visual SLAM.

PubMed

Mahmoud, Nader; Grasa, Óscar G; Nicolau, Stéphane A; Doignon, Christophe; Soler, Luc; Marescaux, Jacques; Montiel, J M M

2017-01-01

An augmented reality system to visualize a 3D preoperative anatomical model on intra-operative patient is proposed. The hardware requirement is commercial tablet-PC equipped with a camera. Thus, no external tracking device nor artificial landmarks on the patient are required. We resort to visual SLAM to provide markerless real-time tablet-PC camera location with respect to the patient. The preoperative model is registered with respect to the patient through 4-6 anchor points. The anchors correspond to anatomical references selected on the tablet-PC screen at the beginning of the procedure. Accurate and real-time preoperative model alignment (approximately 5-mm mean FRE and TRE) was achieved, even when anchors were not visible in the current field of view. The system has been experimentally validated on human volunteers, in vivo pigs and a phantom. The proposed system can be smoothly integrated into the surgical workflow because it: (1) operates in real time, (2) requires minimal additional hardware only a tablet-PC with camera, (3) is robust to occlusion, (4) requires minimal interaction from the medical staff.

A learning-based markerless approach for full-body kinematics estimation in-natura from a single image.

PubMed

Drory, Ami; Li, Hongdong; Hartley, Richard

2017-04-11

We present a supervised machine learning approach for markerless estimation of human full-body kinematics for a cyclist from an unconstrained colour image. This approach is motivated by the limitations of existing marker-based approaches restricted by infrastructure, environmental conditions, and obtrusive markers. By using a discriminatively learned mixture-of-parts model, we construct a probabilistic tree representation to model the configuration and appearance of human body joints. During the learning stage, a Structured Support Vector Machine (SSVM) learns body parts appearance and spatial relations. In the testing stage, the learned models are employed to recover body pose via searching in a test image over a pyramid structure. We focus on the movement modality of cycling to demonstrate the efficacy of our approach. In natura estimation of cycling kinematics using images is challenging because of human interaction with a bicycle causing frequent occlusions. We make no assumptions in relation to the kinematic constraints of the model, nor the appearance of the scene. Our technique finds multiple quality hypotheses for the pose. We evaluate the precision of our method on two new datasets using loss functions. Our method achieves a score of 91.1 and 69.3 on mean Probability of Correct Keypoint (PCK) measure and 88.7 and 66.1 on the Average Precision of Keypoints (APK) measure for the frontal and sagittal datasets respectively. We conclude that our method opens new vistas to robust user-interaction free estimation of full body kinematics, a prerequisite to motion analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Real time markerless motion tracking using linked kinematic chains

DOEpatents

Luck, Jason P [Arvada, CO; Small, Daniel E [Albuquerque, NM

2007-08-14

A markerless method is described for tracking the motion of subjects in a three dimensional environment using a model based on linked kinematic chains. The invention is suitable for tracking robotic, animal or human subjects in real-time using a single computer with inexpensive video equipment, and does not require the use of markers or specialized clothing. A simple model of rigid linked segments is constructed of the subject and tracked using three dimensional volumetric data collected by a multiple camera video imaging system. A physics based method is then used to compute forces to align the model with subsequent volumetric data sets in real-time. The method is able to handle occlusion of segments and accommodates joint limits, velocity constraints, and collision constraints and provides for error recovery. The method further provides for elimination of singularities in Jacobian based calculations, which has been problematic in alternative methods.

Visual tracking of da Vinci instruments for laparoscopic surgery

NASA Astrophysics Data System (ADS)

Speidel, S.; Kuhn, E.; Bodenstedt, S.; Röhl, S.; Kenngott, H.; Müller-Stich, B.; Dillmann, R.

2014-03-01

Intraoperative tracking of laparoscopic instruments is a prerequisite to realize further assistance functions. Since endoscopic images are always available, this sensor input can be used to localize the instruments without special devices or robot kinematics. In this paper, we present an image-based markerless 3D tracking of different da Vinci instruments in near real-time without an explicit model. The method is based on different visual cues to segment the instrument tip, calculates a tip point and uses a multiple object particle filter for tracking. The accuracy and robustness is evaluated with in vivo data.

Markerless positional verification using template matching and triangulation of kV images acquired during irradiation for lung tumors treated in breath-hold

NASA Astrophysics Data System (ADS)

Hazelaar, Colien; Dahele, Max; Mostafavi, Hassan; van der Weide, Lineke; Slotman, Ben; Verbakel, Wilko

2018-06-01

Lung tumors treated in breath-hold are subject to inter- and intra-breath-hold variations, which makes tumor position monitoring during each breath-hold important. A markerless technique is desirable, but limited tumor visibility on kV images makes this challenging. We evaluated if template matching  +  triangulation of kV projection images acquired during breath-hold stereotactic treatments could determine 3D tumor position. Band-pass filtering and/or digital tomosynthesis (DTS) were used as image pre-filtering/enhancement techniques. On-board kV images continuously acquired during volumetric modulated arc irradiation of (i) a 3D-printed anthropomorphic thorax phantom with three lung tumors (n  =  6 stationary datasets, n  =  2 gradually moving), and (ii) four patients (13 datasets) were analyzed. 2D reference templates (filtered DRRs) were created from planning CT data. Normalized cross-correlation was used for 2D matching between templates and pre-filtered/enhanced kV images. For 3D verification, each registration was triangulated with multiple previous registrations. Generally applicable image processing/algorithm settings for lung tumors in breath-hold were identified. For the stationary phantom, the interquartile range of the 3D position vector was on average 0.25 mm for 12° DTS  +  band-pass filtering (average detected positions in 2D  =  99.7%, 3D  =  96.1%, and 3D excluding first 12° due to triangulation angle  =  99.9%) compared to 0.81 mm for band-pass filtering only (55.8/52.9/55.0%). For the moving phantom, RMS errors for the lateral/longitudinal/vertical direction after 12° DTS  +  band-pass filtering were 1.5/0.4/1.1 mm and 2.2/0.3/3.2 mm. For the clinical data, 2D position was determined for at least 93% of each dataset and 3D position excluding first 12° for at least 82% of each dataset using 12° DTS  +  band-pass filtering. Template matching  +  triangulation using DTS  +  band-pass filtered images could accurately determine the position of stationary lung tumors. However, triangulation was less accurate/reliable for targets with continuous, gradual displacement in the lateral and vertical directions. This technique is therefore currently most suited to detect/monitor offsets occurring between initial setup and the start of treatment, inter-breath-hold variations, and tumors with predominantly longitudinal motion.

3D interactive augmented reality-enhanced digital learning systems for mobile devices

NASA Astrophysics Data System (ADS)

Feng, Kai-Ten; Tseng, Po-Hsuan; Chiu, Pei-Shuan; Yang, Jia-Lin; Chiu, Chun-Jie

2013-03-01

With enhanced processing capability of mobile platforms, augmented reality (AR) has been considered a promising technology for achieving enhanced user experiences (UX). Augmented reality is to impose virtual information, e.g., videos and images, onto a live-view digital display. UX on real-world environment via the display can be e ectively enhanced with the adoption of interactive AR technology. Enhancement on UX can be bene cial for digital learning systems. There are existing research works based on AR targeting for the design of e-learning systems. However, none of these work focuses on providing three-dimensional (3-D) object modeling for en- hanced UX based on interactive AR techniques. In this paper, the 3-D interactive augmented reality-enhanced learning (IARL) systems will be proposed to provide enhanced UX for digital learning. The proposed IARL systems consist of two major components, including the markerless pattern recognition (MPR) for 3-D models and velocity-based object tracking (VOT) algorithms. Realistic implementation of proposed IARL system is conducted on Android-based mobile platforms. UX on digital learning can be greatly improved with the adoption of proposed IARL systems.

Validation of 2 noninvasive, markerless reconstruction techniques in biplane high-speed fluoroscopy for 3-dimensional research of bovine distal limb kinematics.

PubMed

Weiss, M; Reich, E; Grund, S; Mülling, C K W; Geiger, S M

2017-10-01

Lameness severely impairs cattle's locomotion, and it is among the most important threats to animal welfare, performance, and productivity in the modern dairy industry. However, insight into the pathological alterations of claw biomechanics leading to lameness and an understanding of the biomechanics behind development of claw lesions causing lameness are limited. Biplane high-speed fluoroscopic kinematography is a new approach for the analysis of skeletal motion. Biplane high-speed videos in combination with bone scans can be used for 3-dimensional (3D) animations of bones moving in 3D space. The gold standard, marker-based animation, requires implantation of radio-opaque markers into bones, which impairs the practicability for lameness research in live animals. Therefore, the purpose of this study was to evaluate the comparative accuracy of 2 noninvasive, markerless animation techniques (semi-automatic and manual) in 3D animation of the bovine distal limb. Tantalum markers were implanted into each of the distal, middle, and proximal phalanges of 5 isolated bovine distal forelimbs, and biplane high-speed x-ray videos of each limb were recorded to capture the simulation of one step. The limbs were scanned by computed tomography to create bone models of the 6 digital bones, and 3D animation of the bones' movements were subsequently reconstructed using the marker-based, the semi-automatic, and the manual animation techniques. Manual animation translational bias and precision varied from 0.63 ± 0.26 mm to 0.80 ± 0.49 mm, and rotational bias and precision ranged from 2.41 ± 1.43° to 6.75 ± 4.67°. Semi-automatic translational values for bias and precision ranged from 1.26 ± 1.28 mm to 2.75 ± 2.17 mm, and rotational values varied from 3.81 ± 2.78° to 11.7 ± 8.11°. In our study, we demonstrated the successful application of biplane high-speed fluoroscopic kinematography to gait analysis of bovine distal limb. Using the manual animation technique, kinematics can be measured with sub-millimeter accuracy without the need for invasive marker implantation. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Evaluation of Simulated Clinical Breast Exam Motion Patterns Using Marker-Less Video Tracking

PubMed Central

Azari, David P.; Pugh, Carla M.; Laufer, Shlomi; Kwan, Calvin; Chen, Chia-Hsiung; Yen, Thomas Y.; Hu, Yu Hen; Radwin, Robert G.

2016-01-01

Objective This study investigates using marker-less video tracking to evaluate hands-on clinical skills during simulated clinical breast examinations (CBEs). Background There are currently no standardized and widely accepted CBE screening techniques. Methods Experienced physicians attending a national conference conducted simulated CBEs presenting different pathologies with distinct tumorous lesions. Single hand exam motion was recorded and analyzed using marker-less video tracking. Four kinematic measures were developed to describe temporal (time pressing and time searching) and spatial (area covered and distance explored) patterns. Results Mean differences between time pressing, area covered, and distance explored varied across the simulated lesions. Exams were objectively categorized as either sporadic, localized, thorough, or efficient for both temporal and spatial categories based on spatiotemporal characteristics. The majority of trials were temporally or spatially thorough (78% and 91%), exhibiting proportionally greater time pressing and time searching (temporally thorough) and greater area probed with greater distance explored (spatially thorough). More efficient exams exhibited proportionally more time pressing with less time searching (temporally efficient) and greater area probed with less distance explored (spatially efficient). Just two (5.9 %) of the trials exhibited both high temporal and spatial efficiency. Conclusions Marker-less video tracking was used to discriminate different examination techniques and measure when an exam changes from general searching to specific probing. The majority of participants exhibited more thorough than efficient patterns. Application Marker-less video kinematic tracking may be useful for quantifying clinical skills for training and assessment. PMID:26546381

Evaluation of Hands-On Clinical Exam Performance Using Marker-less Video Tracking.

PubMed

Azari, David; Pugh, Carla; Laufer, Shlomi; Cohen, Elaine; Kwan, Calvin; Chen, Chia-Hsiung Eric; Yen, Thomas Y; Hu, Yu Hen; Radwin, Robert

2014-09-01

This study investigates the potential of using marker-less video tracking of the hands for evaluating hands-on clinical skills. Experienced family practitioners attending a national conference were recruited and asked to conduct a breast examination on a simulator that simulates different clinical presentations. Videos were made of the clinician's hands during the exam and video processing software for tracking hand motion to quantify hand motion kinematics was used. Practitioner motion patterns indicated consistent behavior of participants across multiple pathologies. Different pathologies exhibited characteristic motion patterns in the aggregate at specific parts of an exam, indicating consistent inter-participant behavior. Marker-less video kinematic tracking therefore shows promise in discriminating between different examination procedures, clinicians, and pathologies.

Establishment of a Cre recombinase based mutagenesis protocol for markerless gene deletion in Streptococcus suis.

PubMed

Koczula, A; Willenborg, J; Bertram, R; Takamatsu, D; Valentin-Weigand, P; Goethe, R

2014-12-01

The lack of knowledge about pathogenicity mechanisms of Streptococcus (S.) suis is, at least partially, attributed to limited methods for its genetic manipulation. Here, we established a Cre-lox based recombination system for markerless gene deletions in S. suis serotype 2 with high selective pressure and without undesired side effects. Copyright © 2014 Elsevier B.V. All rights reserved.

Recombineering in Streptococcus mutans Using Direct Repeat-Mediated Cloning-Independent Markerless Mutagenesis (DR-CIMM).

PubMed

Zhang, Shan; Zou, Zhengzhong; Kreth, Jens; Merritt, Justin

2017-01-01

Studies of the dental caries pathogen Streptococcus mutans have benefitted tremendously from its sophisticated genetic system. As part of our own efforts to further improve upon the S. mutans genetic toolbox, we previously reported the development of the first cloning-independent markerless mutagenesis (CIMM) system for S. mutans and illustrated how this approach could be adapted for use in many other organisms. The CIMM approach only requires overlap extension PCR (OE-PCR) protocols to assemble counterselectable allelic replacement mutagenesis constructs, and thus greatly increased the speed and efficiency with which markerless mutations could be introduced into S. mutans . Despite its utility, the system is still subject to a couple limitations. Firstly, CIMM requires negative selection with the conditionally toxic phenylalanine analog p -chlorophenylalanine (4-CP), which is efficient, but never perfect. Typically, 4-CP negative selection results in a small percentage of naturally resistant background colonies. Secondly, CIMM requires two transformation steps to create markerless mutants. This can be inherently problematic if the transformability of the strain is negatively impacted after the first transformation step, which is used to insert the counterselection cassette at the mutation site on the chromosome. In the current study, we develop a next-generation counterselection cassette that eliminates 4-CP background resistance and combine this with a new direct repeat-mediated cloning-independent markerless mutagenesis (DR-CIMM) system to specifically address the limitations of the prior approach. DR-CIMM is even faster and more efficient than CIMM for the creation of all types of deletions, insertions, and point mutations and is similarly adaptable for use in a wide range of genetically tractable bacteria.

Virtual Exercise Training Software System

NASA Technical Reports Server (NTRS)

Vu, L.; Kim, H.; Benson, E.; Amonette, W. E.; Barrera, J.; Perera, J.; Rajulu, S.; Hanson, A.

2018-01-01

The purpose of this study was to develop and evaluate a virtual exercise training software system (VETSS) capable of providing real-time instruction and exercise feedback during exploration missions. A resistive exercise instructional system was developed using a Microsoft Kinect depth-camera device, which provides markerless 3-D whole-body motion capture at a small form factor and minimal setup effort. It was hypothesized that subjects using the newly developed instructional software tool would perform the deadlift exercise with more optimal kinematics and consistent technique than those without the instructional software. Following a comprehensive evaluation in the laboratory, the system was deployed for testing and refinement in the NASA Extreme Environment Mission Operations (NEEMO) analog.

Development of a Markerless Genetic Exchange System in Desulfovibrio vulgaris Hildenborough and Its Use in Generating a Strain with Increased Transformation Efficiency

DOE Office of Scientific and Technical Information (OSTI.GOV)

Keller, Kimberly L.; Bender, Kelly S.; Wall, Judy D.

2009-07-21

In recent years, the genetic manipulation of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough has seen enormous progress. In spite of this progress, the current marker exchange deletion method does not allow for easy selection of multiple sequential gene deletions in a single strain because of the limited number of selectable markers available in D. vulgaris. To broaden the repertoire of genetic tools for manipulation, an in-frame, markerless deletion system has been developed. The counterselectable marker that makes this deletion system possible is the pyrimidine salvage enzyme, uracil phosphoribosyltransferase, encoded by upp. In wild-type D. vulgaris, growth was shown to bemore » inhibited by the toxic pyrimidine analog 5-fluorouracil (5-FU); whereas, a mutant bearing a deletion of the upp gene was resistant to 5-FU. When a plasmid containing the wild-type upp gene expressed constitutively from the aph(3')-II promoter (promoter for the kanamycin resistance gene in Tn5) was introduced into the upp deletion strain, sensitivity to 5-FU was restored. This observation allowed us to develop a two-step integration and excision strategy for the deletion of genes of interest. Since this inframe deletion strategy does not retain an antibiotic cassette, multiple deletions can be generated in a single strain without the accumulation of genes conferring antibiotic resistances. We used this strategy to generate a deletion strain lacking the endonuclease (hsdR, DVU1703) of a type I restriction-modification system, that we designated JW7035. The transformation efficiency of the JW7035 strain was found to be 100 to 1000 times greater than that of the wild-type strain when stable plasmids were introduced via electroporation.« less

Markerless rat head motion tracking using structured light for brain PET imaging of unrestrained awake small animals

NASA Astrophysics Data System (ADS)

Miranda, Alan; Staelens, Steven; Stroobants, Sigrid; Verhaeghe, Jeroen

2017-03-01

Preclinical positron emission tomography (PET) imaging in small animals is generally performed under anesthesia to immobilize the animal during scanning. More recently, for rat brain PET studies, methods to perform scans of unrestrained awake rats are being developed in order to avoid the unwanted effects of anesthesia on the brain response. Here, we investigate the use of a projected structure stereo camera to track the motion of the rat head during the PET scan. The motion information is then used to correct the PET data. The stereo camera calculates a 3D point cloud representation of the scene and the tracking is performed by point cloud matching using the iterative closest point algorithm. The main advantage of the proposed motion tracking is that no intervention, e.g. for marker attachment, is needed. A manually moved microDerenzo phantom experiment and 3 awake rat [18F]FDG experiments were performed to evaluate the proposed tracking method. The tracking accuracy was 0.33 mm rms. After motion correction image reconstruction, the microDerenzo phantom was recovered albeit with some loss of resolution. The reconstructed FWHM of the 2.5 and 3 mm rods increased with 0.94 and 0.51 mm respectively in comparison with the motion-free case. In the rat experiments, the average tracking success rate was 64.7%. The correlation of relative brain regional [18F]FDG uptake between the anesthesia and awake scan reconstructions was increased from on average 0.291 (not significant) before correction to 0.909 (p  <  0.0001) after motion correction. Markerless motion tracking using structured light can be successfully used for tracking of the rat head for motion correction in awake rat PET scans.

A markerless system based on smartphones and webcam for the measure of step length, width and duration on treadmill.

PubMed

Barone, V; Verdini, F; Burattini, L; Di Nardo, F; Fioretti, S

2016-03-01

A markerless low cost prototype has been developed for the determination of some spatio-temporal parameters of human gait: step-length, step-width and cadence have been considered. Only a smartphone and a high-definition webcam have been used. The signals obtained by the accelerometer embedded in the smartphone are used to recognize the heel strike events, while the feet positions are calculated through image processing of the webcam stream. Step length and width are computed during gait trials on a treadmill at various speeds (3, 4 and 5 km/h). Six subjects have been tested for a total of 504 steps. Results were compared with those obtained by a stereo-photogrammetric system (Elite, BTS Engineering). The maximum average errors were 3.7 cm (5.36%) for the right step length and 1.63 cm (15.16%) for the right step width at 5 km/h. The maximum average error for step duration was 0.02 s (1.69%) at 5 km/h for the right steps. The system is characterized by a very high level of automation that allows its use by non-expert users in non-structured environments. A low cost system able to automatically provide a reliable and repeatable evaluation of some gait events and parameters during treadmill walking, is relevant also from a clinical point of view because it allows the analysis of hundreds of steps and consequently an analysis of their variability. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

WE-AB-303-08: Direct Lung Tumor Tracking Using Short Imaging Arcs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shieh, C; Huang, C; Keall, P

2015-06-15

Purpose: Most current tumor tracking technologies rely on implanted markers, which suffer from potential toxicity of marker placement and mis-targeting due to marker migration. Several markerless tracking methods have been proposed: these are either indirect methods or have difficulties tracking lung tumors in most clinical cases due to overlapping anatomies in 2D projection images. We propose a direct lung tumor tracking algorithm robust to overlapping anatomies using short imaging arcs. Methods: The proposed algorithm tracks the tumor based on kV projections acquired within the latest six-degree imaging arc. To account for respiratory motion, an external motion surrogate is used tomore » select projections of the same phase within the latest arc. For each arc, the pre-treatment 4D cone-beam CT (CBCT) with tumor contours are used to estimate and remove the contribution to the integral attenuation from surrounding anatomies. The position of the tumor model extracted from 4D CBCT of the same phase is then optimized to match the processed projections using the conjugate gradient method. The algorithm was retrospectively validated on two kV scans of a lung cancer patient with implanted fiducial markers. This patient was selected as the tumor is attached to the mediastinum, representing a challenging case for markerless tracking methods. The tracking results were converted to expected marker positions and compared with marker trajectories obtained via direct marker segmentation (ground truth). Results: The root-mean-squared-errors of tracking were 0.8 mm and 0.9 mm in the superior-inferior direction for the two scans. Tracking error was found to be below 2 and 3 mm for 90% and 98% of the time, respectively. Conclusions: A direct lung tumor tracking algorithm robust to overlapping anatomies was proposed and validated on two scans of a lung cancer patient. Sub-millimeter tracking accuracy was observed, indicating the potential of this algorithm for real-time guidance applications.« less

Image overlay navigation by markerless surface registration in gastrointestinal, hepatobiliary and pancreatic surgery.

PubMed

Sugimoto, Maki; Yasuda, Hideki; Koda, Keiji; Suzuki, Masato; Yamazaki, Masato; Tezuka, Tohru; Kosugi, Chihiro; Higuchi, Ryota; Watayo, Yoshihisa; Yagawa, Yohsuke; Uemura, Shuichiro; Tsuchiya, Hironori; Azuma, Takeshi

2010-09-01

We applied a new concept of "image overlay surgery" consisting of the integration of virtual reality (VR) and augmented reality (AR) technology, in which dynamic 3D images were superimposed on the patient's actual body surface and evaluated as a reference for surgical navigation in gastrointestinal, hepatobiliary and pancreatic surgery. We carried out seven surgeries, including three cholecystectomies, two gastrectomies and two colectomies. A Macintosh and a DICOM workstation OsiriX were used in the operating room for image analysis. Raw data of the preoperative patient information obtained via MDCT were reconstructed to volume rendering and projected onto the patient's body surface during the surgeries. For accurate registration, OsiriX was first set to reproduce the patient body surface, and the positional coordinates of the umbilicus, left and right nipples, and the inguinal region were fixed as physiological markers on the body surface to reduce the positional error. The registration process was non-invasive and markerlesss, and was completed within 5 min. Image overlay navigation was helpful for 3D anatomical understanding of the surgical target in the gastrointestinal, hepatobiliary and pancreatic anatomies. The surgeon was able to minimize movement of the gaze and could utilize the image assistance without interfering with the forceps operation, reducing the gap from the VR. Unexpected organ injury could be avoided in all procedures. In biliary surgery, the projected virtual cholangiogram on the abdominal wall could advance safely with identification of the bile duct. For early gastric and colorectal cancer, the small tumors and blood vessels, which usually could not be found on the gastric serosa by laparoscopic view, were simultaneously detected on the body surface by carbon dioxide-enhanced MDCT. This provided accurate reconstructions of the tumor and involved lymph node, directly linked with optimization of the surgical procedures. Our non-invasive markerless registration using physiological markers on the body surface reduced logistical efforts. The image overlay technique is a useful tool when highlighting hidden structures, giving more information.

MIT-Skywalker: On the use of a markerless system.

PubMed

Goncalves, Rogerio S; Hamilton, Taya; Krebs, Hermano I

2017-07-01

This paper describes our efforts to employ the Microsoft Kinect as a low cost vision control system for the MIT-Skywalker, a robotic gait rehabilitation device. The Kinect enables an alternative markerless solution to control the MIT-Skywalker and allows a more user-friendly set-up. A study involving eight healthy subjects and two stroke survivors using the MIT-Skywalker device demonstrates the advantages and challenges of this new proposed approach.

Marker-less respiratory motion modeling using the Microsoft Kinect for Windows

NASA Astrophysics Data System (ADS)

Tahavori, F.; Alnowami, M.; Wells, K.

2014-03-01

Patient respiratory motion is a major problem during external beam radiotherapy of the thoracic and abdominal regions due to the associated organ and target motion. In addition, such motion introduces uncertainty in both radiotherapy planning and delivery and may potentially vary between the planning and delivery sessions. The aim of this work is to examine subject-specific external respiratory motion and its associated drift from an assumed average cycle which is the basis for many respiratory motion compensated applications including radiotherapy treatment planning and delivery. External respiratory motion data were acquired from a group of 20 volunteers using a marker-less 3D depth camera, Kinect for Windows. The anterior surface encompassing thoracic and abdominal regions were subject to principal component analysis (PCA) to investigate dominant variations. The first principal component typically describes more than 70% of the motion data variance in the thoracic and abdominal surfaces. Across all of the subjects used in this study, 58% of subjects demonstrate largely abdominal breathing and 33% exhibited largely thoracic dominated breathing. In most cases there is observable drift in respiratory motion during the 300s capture period, which is visually demonstrated using Kernel Density Estimation. This study demonstrates that for this cohort of apparently healthy volunteers, there is significant respiratory motion drift in most cases, in terms of amplitude and relative displacement between the thoracic and abdominal respiratory components. This has implications for the development of effective motion compensation methodology.

Feasibility Study for Markerless Tracking of Lung Tumors in Stereotactic Body Radiotherapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Richter, Anne, E-mail: richter_a3@klinik.uni-wuerzburg.d; Wilbert, Juergen; Baier, Kurt

2010-10-01

Purpose: To evaluate the feasibility and accuracy of a method for markerless tracking of lung tumors in electronic portal imaging device (EPID) movies and to analyze intra- and interfractional variations in tumor motion. Methods and Materials: EPID movies were acquired during stereotactic body radiotherapy (SBRT) given to 40 patients with 49 pulmonary targets and retrospectively analyzed. Tumor visibility and tracking accuracy were determined by three observers. Tumor motion of 30 targets was analyzed in detail via four-dimensional computed tomography (4DCT) and EPID in the superior-inferior direction for intra- and interfractional variations. Results: Tumor visibility was sufficient for markerless tracking inmore » 47% of the EPID movies. Tumor size and visibility in the DRR were correlated with visibility in the EPID images. The difference between automatic and manual tracking was a maximum of 2 mm for 98.3% in the x direction and 89.4% in the y direction. Motion amplitudes in 4DCT images (range, 0.7-17.9 mm; median, 4.9 mm) were closely correlated with amplitudes in the EPID movies. Intrafractional and interfractional variability of tumor motion amplitude were of similar magnitude: 1 mm on average to a maximum of 4 mm. A change in moving average of more than {+-}1 mm, {+-}2 mm, and {+-}4 mm were observed in 47.1%, 17.1%, and 4.5% of treatment time for all trajectories, respectively. Mean tumor velocity was 3.4 mm/sec, to a maximum 61 mm/sec. Conclusions: Tracking of pulmonary tumors in EPID images without implanted markers was feasible in 47% of all treatment beams. 4DCT is representative of the evaluation of mean breathing motion on average, but larger deviations occurred in target motion between treatment planning and delivery effort a monitoring during delivery.« less

D Building Reconstruction by Multiview Images and the Integrated Application with Augmented Reality

NASA Astrophysics Data System (ADS)

Hwang, Jin-Tsong; Chu, Ting-Chen

2016-10-01

This study presents an approach wherein photographs with a high degree of overlap are clicked using a digital camera and used to generate three-dimensional (3D) point clouds via feature point extraction and matching. To reconstruct a building model, an unmanned aerial vehicle (UAV) is used to click photographs from vertical shooting angles above the building. Multiview images are taken from the ground to eliminate the shielding effect on UAV images caused by trees. Point clouds from the UAV and multiview images are generated via Pix4Dmapper. By merging two sets of point clouds via tie points, the complete building model is reconstructed. The 3D models are reconstructed using AutoCAD 2016 to generate vectors from the point clouds; SketchUp Make 2016 is used to rebuild a complete building model with textures. To apply 3D building models in urban planning and design, a modern approach is to rebuild the digital models; however, replacing the landscape design and building distribution in real time is difficult as the frequency of building replacement increases. One potential solution to these problems is augmented reality (AR). Using Unity3D and Vuforia to design and implement the smartphone application service, a markerless AR of the building model can be built. This study is aimed at providing technical and design skills related to urban planning, urban designing, and building information retrieval using AR.

The Accuracy of Conventional 2D Video for Quantifying Upper Limb Kinematics in Repetitive Motion Occupational Tasks

PubMed Central

Chen, Chia-Hsiung; Azari, David; Hu, Yu Hen; Lindstrom, Mary J.; Thelen, Darryl; Yen, Thomas Y.; Radwin, Robert G.

2015-01-01

Objective Marker-less 2D video tracking was studied as a practical means to measure upper limb kinematics for ergonomics evaluations. Background Hand activity level (HAL) can be estimated from speed and duty cycle. Accuracy was measured using a cross correlation template-matching algorithm for tracking a region of interest on the upper extremities. Methods Ten participants performed a paced load transfer task while varying HAL (2, 4, and 5) and load (2.2 N, 8.9 N and 17.8 N). Speed and acceleration measured from 2D video were compared against ground truth measurements using 3D infrared motion capture. Results The median absolute difference between 2D video and 3D motion capture was 86.5 mm/s for speed, and 591 mm/s2 for acceleration, and less than 93 mm/s for speed and 656 mm/s2 for acceleration when camera pan and tilt were within ±30 degrees. Conclusion Single-camera 2D video had sufficient accuracy (< 100 mm/s) for evaluating HAL. Practitioner Summary This study demonstrated that 2D video tracking had sufficient accuracy to measure HAL for ascertaining the American Conference of Government Industrial Hygienists Threshold Limit Value® for repetitive motion when the camera is located within ±30 degrees off the plane of motion when compared against 3D motion capture for a simulated repetitive motion task. PMID:25978764

[Regulation of sporulation by two-component system YvcPQ in Bacillus thuringiensis].

PubMed

Fan, Qingyun; Zhang, Shumeng; Gong, Yujing; He, Jin

2017-01-04

To study the regulation of sporulation controlled by two-component system (TCS) YvcPQ. β-galactosidase experiment was used to verify the regulation of YvcP on kapD expression; bacterial one-hybrid assay, EMSA and RT-qPCR were applied to study the regulation of AbrB on yvcPQ expression; markerless gene deletion coupled with spore count was used to reveal the influence of yvcPQ and kapD expressions on sporulation. transcriptional regulator AbrB up-regulated the expression of yvcPQ; YvcP promoted the expression of kapD to inhibit sporulation. AbrB up-regulated the transcription of yvcPQ operon, then the increased YvcP strengthened the transcriptional acitivation of sporulation inhibitor gene kapD, and subsequently inhibited sporulation.

Real-time soft tissue motion estimation for lung tumors during radiotherapy delivery.

PubMed

Rottmann, Joerg; Keall, Paul; Berbeco, Ross

2013-09-01

To provide real-time lung tumor motion estimation during radiotherapy treatment delivery without the need for implanted fiducial markers or additional imaging dose to the patient. 2D radiographs from the therapy beam's-eye-view (BEV) perspective are captured at a frame rate of 12.8 Hz with a frame grabber allowing direct RAM access to the image buffer. An in-house developed real-time soft tissue localization algorithm is utilized to calculate soft tissue displacement from these images in real-time. The system is tested with a Varian TX linear accelerator and an AS-1000 amorphous silicon electronic portal imaging device operating at a resolution of 512 × 384 pixels. The accuracy of the motion estimation is verified with a dynamic motion phantom. Clinical accuracy was tested on lung SBRT images acquired at 2 fps. Real-time lung tumor motion estimation from BEV images without fiducial markers is successfully demonstrated. For the phantom study, a mean tracking error <1.0 mm [root mean square (rms) error of 0.3 mm] was observed. The tracking rms accuracy on BEV images from a lung SBRT patient (≈20 mm tumor motion range) is 1.0 mm. The authors demonstrate for the first time real-time markerless lung tumor motion estimation from BEV images alone. The described system can operate at a frame rate of 12.8 Hz and does not require prior knowledge to establish traceable landmarks for tracking on the fly. The authors show that the geometric accuracy is similar to (or better than) previously published markerless algorithms not operating in real-time.

SU-E-J-188: Theoretical Estimation of Margin Necessary for Markerless Motion Tracking

DOE Office of Scientific and Technical Information (OSTI.GOV)

Patel, R; Block, A; Harkenrider, M

2015-06-15

Purpose: To estimate the margin necessary to adequately cover the target using markerless motion tracking (MMT) of lung lesions given the uncertainty in tracking and the size of the target. Methods: Simulations were developed in Matlab to determine the effect of tumor size and tracking uncertainty on the margin necessary to achieve adequate coverage of the target. For simplicity, the lung tumor was approximated by a circle on a 2D radiograph. The tumor was varied in size from a diameter of 0.1 − 30 mm in increments of 0.1 mm. From our previous studies using dual energy markerless motion tracking,more » we estimated tracking uncertainties in x and y to have a standard deviation of 2 mm. A Gaussian was used to simulate the deviation between the tracked location and true target location. For each size tumor, 100,000 deviations were randomly generated, the margin necessary to achieve at least 95% coverage 95% of the time was recorded. Additional simulations were run for varying uncertainties to demonstrate the effect of the tracking accuracy on the margin size. Results: The simulations showed an inverse relationship between tumor size and margin necessary to achieve 95% coverage 95% of the time using the MMT technique. The margin decreased exponentially with target size. An increase in tracking accuracy expectedly showed a decrease in margin size as well. Conclusion: In our clinic a 5 mm expansion of the internal target volume (ITV) is used to define the planning target volume (PTV). These simulations show that for tracking accuracies in x and y better than 2 mm, the margin required is less than 5 mm. This simple simulation can provide physicians with a guideline estimation for the margin necessary for use of MMT clinically based on the accuracy of their tracking and the size of the tumor.« less

Automatic human body modeling for vision-based motion capture system using B-spline parameterization of the silhouette

NASA Astrophysics Data System (ADS)

Jaume-i-Capó, Antoni; Varona, Javier; González-Hidalgo, Manuel; Mas, Ramon; Perales, Francisco J.

2012-02-01

Human motion capture has a wide variety of applications, and in vision-based motion capture systems a major issue is the human body model and its initialization. We present a computer vision algorithm for building a human body model skeleton in an automatic way. The algorithm is based on the analysis of the human shape. We decompose the body into its main parts by computing the curvature of a B-spline parameterization of the human contour. This algorithm has been applied in a context where the user is standing in front of a camera stereo pair. The process is completed after the user assumes a predefined initial posture so as to identify the main joints and construct the human model. Using this model, the initialization problem of a vision-based markerless motion capture system of the human body is solved.

Optical fringe-reflection deflectometry with bundle adjustment

NASA Astrophysics Data System (ADS)

Xiao, Yong-Liang; Li, Sikun; Zhang, Qican; Zhong, Jianxin; Su, Xianyu; You, Zhisheng

2018-06-01

Liquid crystal display (LCD) screens are located outside of a camera's field of view in fringe-reflection deflectometry. Therefore, fringes that are displayed on LCD screens are obtained through specular reflection by a fixed camera. Thus, the pose calibration between the camera and LCD screen is one of the main challenges in fringe-reflection deflectometry. A markerless planar mirror is used to reflect the LCD screen more than three times, and the fringes are mapped into the fixed camera. The geometrical calibration can be accomplished by estimating the pose between the camera and the virtual image of fringes. Considering the relation between their pose, the incidence and reflection rays can be unified in the camera frame, and a forward triangulation intersection can be operated in the camera frame to measure three-dimensional (3D) coordinates of the specular surface. In the final optimization, constraint-bundle adjustment is operated to refine simultaneously the camera intrinsic parameters, including distortion coefficients, estimated geometrical pose between the LCD screen and camera, and 3D coordinates of the specular surface, with the help of the absolute phase collinear constraint. Simulation and experiment results demonstrate that the pose calibration with planar mirror reflection is simple and feasible, and the constraint-bundle adjustment can enhance the 3D coordinate measurement accuracy in fringe-reflection deflectometry.

Markerless 3D motion capture for animal locomotion studies

PubMed Central

Sellers, William Irvin; Hirasaki, Eishi

2014-01-01

ABSTRACT Obtaining quantitative data describing the movements of animals is an essential step in understanding their locomotor biology. Outside the laboratory, measuring animal locomotion often relies on video-based approaches and analysis is hampered because of difficulties in calibration and often the limited availability of possible camera positions. It is also usually restricted to two dimensions, which is often an undesirable over-simplification given the essentially three-dimensional nature of many locomotor performances. In this paper we demonstrate a fully three-dimensional approach based on 3D photogrammetric reconstruction using multiple, synchronised video cameras. This approach allows full calibration based on the separation of the individual cameras and will work fully automatically with completely unmarked and undisturbed animals. As such it has the potential to revolutionise work carried out on free-ranging animals in sanctuaries and zoological gardens where ad hoc approaches are essential and access within enclosures often severely restricted. The paper demonstrates the effectiveness of video-based 3D photogrammetry with examples from primates and birds, as well as discussing the current limitations of this technique and illustrating the accuracies that can be obtained. All the software required is open source so this can be a very cost effective approach and provides a methodology of obtaining data in situations where other approaches would be completely ineffective. PMID:24972869

Simple Method for Markerless Gene Deletion in Multidrug-Resistant Acinetobacter baumannii

PubMed Central

Oh, Man Hwan; Lee, Je Chul; Kim, Jungmin

2015-01-01

The traditional markerless gene deletion technique based on overlap extension PCR has been used for generating gene deletions in multidrug-resistant Acinetobacter baumannii. However, the method is time-consuming because it requires restriction digestion of the PCR products in DNA cloning and the construction of new vectors containing a suitable antibiotic resistance cassette for the selection of A. baumannii merodiploids. Moreover, the availability of restriction sites and the selection of recombinant bacteria harboring the desired chimeric plasmid are limited, making the construction of a chimeric plasmid more difficult. We describe a rapid and easy cloning method for markerless gene deletion in A. baumannii, which has no limitation in the availability of restriction sites and allows for easy selection of the clones carrying the desired chimeric plasmid. Notably, it is not necessary to construct new vectors in our method. This method utilizes direct cloning of blunt-end DNA fragments, in which upstream and downstream regions of the target gene are fused with an antibiotic resistance cassette via overlap extension PCR and are inserted into a blunt-end suicide vector developed for blunt-end cloning. Importantly, the antibiotic resistance cassette is placed outside the downstream region in order to enable easy selection of the recombinants carrying the desired plasmid, to eliminate the antibiotic resistance cassette via homologous recombination, and to avoid the necessity of constructing new vectors. This strategy was successfully applied to functional analysis of the genes associated with iron acquisition by A. baumannii ATCC 19606 and to ompA gene deletion in other A. baumannii strains. Consequently, the proposed method is invaluable for markerless gene deletion in multidrug-resistant A. baumannii. PMID:25746991

Real-time soft tissue motion estimation for lung tumors during radiotherapy delivery

PubMed Central

Rottmann, Joerg; Keall, Paul; Berbeco, Ross

2013-01-01

Purpose: To provide real-time lung tumor motion estimation during radiotherapy treatment delivery without the need for implanted fiducial markers or additional imaging dose to the patient. Methods: 2D radiographs from the therapy beam's-eye-view (BEV) perspective are captured at a frame rate of 12.8 Hz with a frame grabber allowing direct RAM access to the image buffer. An in-house developed real-time soft tissue localization algorithm is utilized to calculate soft tissue displacement from these images in real-time. The system is tested with a Varian TX linear accelerator and an AS-1000 amorphous silicon electronic portal imaging device operating at a resolution of 512 × 384 pixels. The accuracy of the motion estimation is verified with a dynamic motion phantom. Clinical accuracy was tested on lung SBRT images acquired at 2 fps. Results: Real-time lung tumor motion estimation from BEV images without fiducial markers is successfully demonstrated. For the phantom study, a mean tracking error <1.0 mm [root mean square (rms) error of 0.3 mm] was observed. The tracking rms accuracy on BEV images from a lung SBRT patient (≈20 mm tumor motion range) is 1.0 mm. Conclusions: The authors demonstrate for the first time real-time markerless lung tumor motion estimation from BEV images alone. The described system can operate at a frame rate of 12.8 Hz and does not require prior knowledge to establish traceable landmarks for tracking on the fly. The authors show that the geometric accuracy is similar to (or better than) previously published markerless algorithms not operating in real-time. PMID:24007146

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chiu, T; Kearney, V; Liu, H

Purpose: Dynamic tumor tracking or motion compensation techniques have proposed to modify beam delivery following lung tumor motion on the flight. Conventional treatment plan QA could be performed in advance since every delivery may be different. Markerless lung tumor tracking using beams eye view EPID images provides a best treatment evaluation mechanism. The purpose of this study is to improve the accuracy of the online markerless lung tumor motion tracking method. Methods: The lung tumor could be located on every frame of MV images during radiation therapy treatment by comparing with corresponding digitally reconstructed radiograph (DRR). A kV-MV CT correspondingmore » curve is applied on planning kV CT to generate MV CT images for patients in order to enhance the similarity between DRRs and MV treatment images. This kV-MV CT corresponding curve was obtained by scanning a same CT electron density phantom by a kV CT scanner and MV scanner (Tomotherapy) or MV CBCT. Two sets of MV DRRs were then generated for tumor and anatomy without tumor as the references to tracking the tumor on beams eye view EPID images. Results: Phantom studies were performed on a Varian TrueBeam linac. MV treatment images were acquired continuously during each treatment beam delivery at 12 gantry angles by iTools. Markerless tumor tracking was applied with DRRs generated from simulated MVCT. Tumors were tracked on every frame of images and compared with expected positions based on programed phantom motion. It was found that the average tracking error were 2.3 mm. Conclusion: This algorithm is capable of detecting lung tumors at complicated environment without implanting markers. It should be noted that the CT data has a slice thickness of 3 mm. This shows the statistical accuracy is better than the spatial accuracy. This project has been supported by a Varian Research Grant.« less

Fast Markerless Tracking for Augmented Reality in Planar Environment

NASA Astrophysics Data System (ADS)

Basori, Ahmad Hoirul; Afif, Fadhil Noer; Almazyad, Abdulaziz S.; AbuJabal, Hamza Ali S.; Rehman, Amjad; Alkawaz, Mohammed Hazim

2015-12-01

Markerless tracking for augmented reality should not only be accurate but also fast enough to provide a seamless synchronization between real and virtual beings. Current reported methods showed that a vision-based tracking is accurate but requires high computational power. This paper proposes a real-time hybrid-based method for tracking unknown environments in markerless augmented reality. The proposed method provides collaboration of vision-based approach with accelerometers and gyroscopes sensors as camera pose predictor. To align the augmentation relative to camera motion, the tracking method is done by substituting feature-based camera estimation with combination of inertial sensors with complementary filter to provide more dynamic response. The proposed method managed to track unknown environment with faster processing time compared to available feature-based approaches. Moreover, the proposed method can sustain its estimation in a situation where feature-based tracking loses its track. The collaboration of sensor tracking managed to perform the task for about 22.97 FPS, up to five times faster than feature-based tracking method used as comparison. Therefore, the proposed method can be used to track unknown environments without depending on amount of features on scene, while requiring lower computational cost.

TH-AB-202-01: Daily Lung Tumor Motion Characterization On EPIDs Using a Markerless Tiling Model

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rozario, T; University of Texas at Dallas, Richardson, TX; Chiu, T

Purpose: Tracking lung tumor motion in real time allows for target dose escalation while simultaneously reducing dose to sensitive structures, thus increasing local control without increasing toxicity. We present a novel intra-fractional markerless lung tumor tracking algorithm using MV treatment beam images acquired during treatment delivery. Strong signals superimposed on the tumor significantly reduced the soft tissue resolution; while different imaging modalities involved introduce global imaging discrepancies. This reduced the comparison accuracies. A simple yet elegant Tiling algorithm is reported to overcome the aforementioned issues. Methods: MV treatment beam images were acquired continuously in beam’s eye view (BEV) by anmore » electronic portal imaging device (EPID) during treatment and analyzed to obtain tumor positions on every frame. Every frame of the MV image was simulated by a composite of two components with separate digitally reconstructed radiographs (DRRs): all non-moving structures and the tumor. This Titling algorithm divides the global composite DRR and the corresponding MV projection into sub-images called tiles. Rigid registration is performed independently on tile-pairs in order to improve local soft tissue resolution. This enables the composite DRR to be transformed accurately to match the MV projection and attain a high correlation value through a pixel-based linear transformation. The highest cumulative correlation for all tile-pairs achieved over a user-defined search range indicates the 2-D coordinates of the tumor location on the MV projection. Results: This algorithm was successfully applied to cine-mode BEV images acquired during two SBRT plans delivered five times with different motion patterns to each of two phantoms. Approximately 15000 beam’s eye view images were analyzed and tumor locations were successfully identified on every projection with a maximum/average error of 1.8 mm / 1.0 mm. Conclusion: Despite the presence of strong anatomical signal overlapping with tumor images, this markerless detection algorithm accurately tracks intrafractional lung tumor motions. This project is partially supported by an Elekta research grant.« less

Three-dimensional skeletal kinematics of the shoulder girdle and forelimb in walking Alligator

PubMed Central

Baier, David B; Gatesy, Stephen M

2013-01-01

Crocodylians occupy a key phylogenetic position for investigations of archosaur locomotor evolution. Compared to the well-studied hindlimb, relatively little is known about the skeletal movements and mechanics of the forelimb. In this study, we employed manual markerless XROMM (X-ray Reconstruction Of Moving Morphology) to measure detailed 3-D kinematics of the shoulder girdle and forelimb bones of American alligators (Alligator mississippiensis) walking on a treadmill. Digital models of the interclavicle, scapulocoracoid, humerus, radius and ulna were created using a 3-D laser scanner. Models were articulated and aligned to simultaneously recorded frames of fluoroscopic and standard light video to reconstruct and measure joint motion. Joint coordinate systems were established for the coracosternal, glenohumeral and elbow joints. Our analysis revealed that the limb joints only account for about half of fore/aft limb excursion; the remaining excursion results from shoulder girdle movements and lateral bending of the vertebral column. Considerable motion of each scapulocoracoid relative to the vertebral column is consistent with coracosternal mobility. The hemisellar design of the glenohumeral joint permits some additional translation, or sliding in the fore-aft plane, but this movement does not have much of an effect on the distal excursion of the bone. PMID:24102540

CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973.

PubMed

Wendt, Kristen E; Ungerer, Justin; Cobb, Ryan E; Zhao, Huimin; Pakrasi, Himadri B

2016-06-23

As autotrophic prokaryotes, cyanobacteria are ideal chassis organisms for sustainable production of various useful compounds. The newly characterized cyanobacterium Synechococcus elongatus UTEX 2973 is a promising candidate for serving as a microbial cell factory because of its unusually rapid growth rate. Here, we seek to develop a genetic toolkit that enables extensive genomic engineering of Synechococcus 2973 by implementing a CRISPR/Cas9 editing system. We targeted the nblA gene because of its important role in biological response to nitrogen deprivation conditions. First, we determined that the Streptococcus pyogenes Cas9 enzyme is toxic in cyanobacteria, and conjugational transfer of stable, replicating constructs containing the cas9 gene resulted in lethality. However, after switching to a vector that permitted transient expression of the cas9 gene, we achieved markerless editing in 100 % of cyanobacterial exconjugants after the first patch. Moreover, we could readily cure the organisms of antibiotic resistance, resulting in a markerless deletion strain. High expression levels of the Cas9 protein in Synechococcus 2973 appear to be toxic and result in cell death. However, introduction of a CRISPR/Cas9 genome editing system on a plasmid backbone that leads to transient cas9 expression allowed for efficient markerless genome editing in a wild type genetic background.

CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973

DOE PAGES

Wendt, Kristen E.; Ungerer, Justin; Cobb, Ryan E.; ...

2016-06-23

As autotrophic prokaryotes, cyanobacteria are ideal chassis organisms for sustainable production of various useful compounds. The newly characterized cyanobacterium Synechococcus elongatus UTEX 2973 is a promising candidate for serving as a microbial cell factory because of its unusually rapid growth rate. Here, we seek to develop a genetic toolkit that enables extensive genomic engineering of Synechococcus 2973 by implementing a CRISPR/Cas9 editing system. We targeted the nblA gene because of its important role in biological response to nitrogen deprivation conditions. First, we determined that the Streptococcus pyogenes Cas9 enzyme is toxic in cyanobacteria, and conjugational transfer of stable, replicating constructsmore » containing the cas9 gene resulted in lethality. However, after switching to a vector that permitted transient expression of the cas9 gene, we achieved markerless editing in 100 % of cyanobacterial exconjugants after the first patch. Moreover, we could readily cure the organisms of antibiotic resistance, resulting in a markerless deletion strain. In conclusion, high expression levels of the Cas9 protein in Synechococcus 2973 appear to be toxic and result in cell death. However, introduction of a CRISPR/Cas9 genome editing system on a plasmid backbone that leads to transient cas9 expression allowed for efficient markerless genome editing in a wild type genetic background.« less

CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wendt, Kristen E.; Ungerer, Justin; Cobb, Ryan E.

As autotrophic prokaryotes, cyanobacteria are ideal chassis organisms for sustainable production of various useful compounds. The newly characterized cyanobacterium Synechococcus elongatus UTEX 2973 is a promising candidate for serving as a microbial cell factory because of its unusually rapid growth rate. Here, we seek to develop a genetic toolkit that enables extensive genomic engineering of Synechococcus 2973 by implementing a CRISPR/Cas9 editing system. We targeted the nblA gene because of its important role in biological response to nitrogen deprivation conditions. First, we determined that the Streptococcus pyogenes Cas9 enzyme is toxic in cyanobacteria, and conjugational transfer of stable, replicating constructsmore » containing the cas9 gene resulted in lethality. However, after switching to a vector that permitted transient expression of the cas9 gene, we achieved markerless editing in 100 % of cyanobacterial exconjugants after the first patch. Moreover, we could readily cure the organisms of antibiotic resistance, resulting in a markerless deletion strain. In conclusion, high expression levels of the Cas9 protein in Synechococcus 2973 appear to be toxic and result in cell death. However, introduction of a CRISPR/Cas9 genome editing system on a plasmid backbone that leads to transient cas9 expression allowed for efficient markerless genome editing in a wild type genetic background.« less

Cre/lox-based multiple markerless gene disruption in the genome of the extreme thermophile Thermus thermophilus.

PubMed

Togawa, Yoichiro; Nunoshiba, Tatsuo; Hiratsu, Keiichiro

2018-02-01

Markerless gene-disruption technology is particularly useful for effective genetic analyses of Thermus thermophilus (T. thermophilus), which have a limited number of selectable markers. In an attempt to develop a novel system for the markerless disruption of genes in T. thermophilus, we applied a Cre/lox system to construct a triple gene disruptant. To achieve this, we constructed two genetic tools, a loxP-htk-loxP cassette and cre-expressing plasmid, pSH-Cre, for gene disruption and removal of the selectable marker by Cre-mediated recombination. We found that the Cre/lox system was compatible with the proliferation of the T. thermophilus HB27 strain at the lowest growth temperature (50 °C), and thus succeeded in establishing a triple gene disruptant, the (∆TTC1454::loxP, ∆TTC1535KpnI::loxP, ∆TTC1576::loxP) strain, without leaving behind a selectable marker. During the process of the sequential disruption of multiple genes, we observed the undesired deletion and inversion of the chromosomal region between multiple loxP sites that were induced by Cre-mediated recombination. Therefore, we examined the effects of a lox66-htk-lox71 cassette by exploiting the mutant lox sites, lox66 and lox71, instead of native loxP sites. We successfully constructed a (∆TTC1535::lox72, ∆TTC1537::lox72) double gene disruptant without inducing the undesired deletion of the 0.7-kbp region between the two directly oriented lox72 sites created by the Cre-mediated recombination of the lox66-htk-lox71 cassette. This is the first demonstration of a Cre/lox system being applicable to extreme thermophiles in a genetic manipulation. Our results indicate that this system is a powerful tool for multiple markerless gene disruption in T. thermophilus.

Robust object tracking techniques for vision-based 3D motion analysis applications

NASA Astrophysics Data System (ADS)

Knyaz, Vladimir A.; Zheltov, Sergey Y.; Vishnyakov, Boris V.

2016-04-01

Automated and accurate spatial motion capturing of an object is necessary for a wide variety of applications including industry and science, virtual reality and movie, medicine and sports. For the most part of applications a reliability and an accuracy of the data obtained as well as convenience for a user are the main characteristics defining the quality of the motion capture system. Among the existing systems for 3D data acquisition, based on different physical principles (accelerometry, magnetometry, time-of-flight, vision-based), optical motion capture systems have a set of advantages such as high speed of acquisition, potential for high accuracy and automation based on advanced image processing algorithms. For vision-based motion capture accurate and robust object features detecting and tracking through the video sequence are the key elements along with a level of automation of capturing process. So for providing high accuracy of obtained spatial data the developed vision-based motion capture system "Mosca" is based on photogrammetric principles of 3D measurements and supports high speed image acquisition in synchronized mode. It includes from 2 to 4 technical vision cameras for capturing video sequences of object motion. The original camera calibration and external orientation procedures provide the basis for high accuracy of 3D measurements. A set of algorithms as for detecting, identifying and tracking of similar targets, so for marker-less object motion capture is developed and tested. The results of algorithms' evaluation show high robustness and high reliability for various motion analysis tasks in technical and biomechanics applications.

Calibration of RGBD camera and cone-beam CT for 3D intra-operative mixed reality visualization.

PubMed

Lee, Sing Chun; Fuerst, Bernhard; Fotouhi, Javad; Fischer, Marius; Osgood, Greg; Navab, Nassir

2016-06-01

This work proposes a novel algorithm to register cone-beam computed tomography (CBCT) volumes and 3D optical (RGBD) camera views. The co-registered real-time RGBD camera and CBCT imaging enable a novel augmented reality solution for orthopedic surgeries, which allows arbitrary views using digitally reconstructed radiographs overlaid on the reconstructed patient's surface without the need to move the C-arm. An RGBD camera is rigidly mounted on the C-arm near the detector. We introduce a calibration method based on the simultaneous reconstruction of the surface and the CBCT scan of an object. The transformation between the two coordinate spaces is recovered using Fast Point Feature Histogram descriptors and the Iterative Closest Point algorithm. Several experiments are performed to assess the repeatability and the accuracy of this method. Target registration error is measured on multiple visual and radio-opaque landmarks to evaluate the accuracy of the registration. Mixed reality visualizations from arbitrary angles are also presented for simulated orthopedic surgeries. To the best of our knowledge, this is the first calibration method which uses only tomographic and RGBD reconstructions. This means that the method does not impose a particular shape of the phantom. We demonstrate a marker-less calibration of CBCT volumes and 3D depth cameras, achieving reasonable registration accuracy. This design requires a one-time factory calibration, is self-contained, and could be integrated into existing mobile C-arms to provide real-time augmented reality views from arbitrary angles.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Poels, Kenneth, E-mail: kenneth.poels@uzbrussel.be; Verellen, Dirk; Van de Vondel, Iwein

Purpose: Because frame rates on current clinical available electronic portal imaging devices (EPID’s) are limited to 7.5 Hz, a new commercially available PerkinElmer EPID (XRD 1642 AP19) with a maximum frame rate of 30 Hz and a new scintillator (Kyokko PI200) with improved sensitivity (light output) for megavolt (MV) irradiation was evaluated. In this work, the influence of MV pulse artifacts and pulsing artifact suppression techniques on fiducial marker and marker-less detection of a lung lesion was investigated, because target localization is an important component of uncertainty in geometrical verification of real-time tumor tracking. Methods: Visicoil™ markers with a diametermore » of 0.05 and 0.075 cm were used for MV marker tracking with a frame rate of, respectively, 7.5, 15, and 30 Hz. A 30 Hz readout of the detector was obtained by a 2 × 2 pixel binning, reducing spatial resolution. Static marker detection was conducted in function of increasing phantom thickness. Additionally, marker-less tracking was conducted and compared with the ground-truth fiducial marker motion. Performance of MV target detection was investigated by comparing the least-square sine wave fit of the detected marker positions with the predefined sine wave motion. For fiducial marker detection, a Laplacian-of-Gaussian enhancement was applied after which normalized cross correlation was used to find the most probable marker position. Marker-less detection was performed by using the scale and orientation adaptive mean shift tracking algorithm. For each MV fluoroscopy, a free running (FR-nF) (ignoring MV pulsing during readout) acquisition mode was compared with two acquisition modes intending to reduce MV pulsing artifacts, i.e., combined wavelet-FFT filtering (FR-wF) and electronic readout synchronized with respect to MV pulses. Results: A 0.05 cm Visicoil marker resulted in an unacceptable root-mean square error (RMSE) > 0.2 cm with a maximum frame rate of 30 Hz during FR-nF readout. With a 30 Hz synchronized readout (S-nF) and during 15 Hz readout (independent of readout mode), RMSE was submillimeter for a static 0.05 cm Visicoil. A dynamic 0.05 cm Visicoil was not detectable on the XRD 1642 AP19, despite a fast synchronized readout. For a 0.075 cm Visicoil, deviations of sine wave motion were submillimeter (RMSE < 0.08 cm), independent of the acquisition mode (FR, S). For marker-less tumor detection, FR-nF images resulted in RMSE > 0.3 cm, while for MV fluoroscopy in S-mode RMSE < 0.1 cm for 15 Hz and RMSE < 0.16 cm for 30 Hz. Largest consistency in target localization was experienced during 15 Hz S-nF readout. Conclusions: In general, marker contrast decreased in function of higher frame rates, which was detrimental for marker detection success. In this work, Visicoils with a thickness of 0.075 cm were showing best results for a 15 Hz frame rate, while non-MV compatible 0.05 cm Visicoil markers were not visible on the new EPID with improved sensitivity compared to EPID models based on a Kodak Lanex Fast scintillator. No noticeable influence of pulsing artifacts on the detection of a 0.075 cm Visicoil was observed, while a synchronized readout provided most reliable detection of a marker-less soft-tissue structure.« less

Dual CRISPR-Cas9 Cleavage Mediated Gene Excision and Targeted Integration in Yarrowia lipolytica.

PubMed

Gao, Difeng; Smith, Spencer; Spagnuolo, Michael; Rodriguez, Gabriel; Blenner, Mark

2018-05-29

CRISPR-Cas9 technology has been successfully applied in Yarrowia lipolytica for targeted genomic editing including gene disruption and integration; however, disruptions by existing methods typically result from small frameshift mutations caused by indels within the coding region, which usually resulted in unnatural protein. In this study, a dual cleavage strategy directed by paired sgRNAs is developed for gene knockout. This method allows fast and robust gene excision, demonstrated on six genes of interest. The targeted regions for excision vary in length from 0.3 kb up to 3.5 kb and contain both non-coding and coding regions. The majority of the gene excisions are repaired by perfect nonhomologous end-joining without indel. Based on this dual cleavage system, two targeted markerless integration methods are developed by providing repair templates. While both strategies are effective, homology mediated end joining (HMEJ) based method are twice as efficient as homology recombination (HR) based method. In both cases, dual cleavage leads to similar or improved gene integration efficiencies compared to gene excision without integration. This dual cleavage strategy will be useful for not only generating more predictable and robust gene knockout, but also for efficient targeted markerless integration, and simultaneous knockout and integration in Y. lipolytica. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Real-Time Augmented Reality System to See-Through Cars.

PubMed

Rameau, Francois; Ha, Hyowon; Joo, Kyungdon; Choi, Jinsoo; Park, Kibaek; Kweon, In So

2016-11-01

One of the most hazardous driving scenario is the overtaking of a slower vehicle, indeed, in this case the front vehicle (being overtaken) can occlude an important part of the field of view of the rear vehicle's driver. This lack of visibility is the most probable cause of accidents in this context. Recent research works tend to prove that augmented reality applied to assisted driving can significantly reduce the risk of accidents. In this paper, we present a real-time marker-less system to see through cars. For this purpose, two cars are equipped with cameras and an appropriate wireless communication system. The stereo vision system mounted on the front car allows to create a sparse 3D map of the environment where the rear car can be localized. Using this inter-car pose estimation, a synthetic image is generated to overcome the occlusion and to create a seamless see-through effect which preserves the structure of the scene.

Video see-through augmented reality for oral and maxillofacial surgery.

PubMed

Wang, Junchen; Suenaga, Hideyuki; Yang, Liangjing; Kobayashi, Etsuko; Sakuma, Ichiro

2017-06-01

Oral and maxillofacial surgery has not been benefitting from image guidance techniques owing to the limitations in image registration. A real-time markerless image registration method is proposed by integrating a shape matching method into a 2D tracking framework. The image registration is performed by matching the patient's teeth model with intraoperative video to obtain its pose. The resulting pose is used to overlay relevant models from the same CT space on the camera video for augmented reality. The proposed system was evaluated on mandible/maxilla phantoms, a volunteer and clinical data. Experimental results show that the target overlay error is about 1 mm, and the frame rate of registration update yields 3-5 frames per second with a 4 K camera. The significance of this work lies in its simplicity in clinical setting and the seamless integration into the current medical procedure with satisfactory response time and overlay accuracy. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Letter regarding 'Comparison between low-cost marker-less and high-end marker-based motion capture systems for the computer-aided assessment of working ergonomics' by Patrizi et al. and research reproducibility.

PubMed

2017-04-01

The reporting of research in a manner that allows reproduction in subsequent investigations is important for scientific progress. Several details of the recent study by Patrizi et al., 'Comparison between low-cost marker-less and high-end marker-based motion capture systems for the computer-aided assessment of working ergonomics', are absent from the published manuscript and make reproduction of findings impossible. As new and complex technologies with great promise for ergonomics develop, new but surmountable challenges for reporting investigations using these technologies in a reproducible manner arise. Practitioner Summary: As with traditional methods, scientific reporting of new and complex ergonomics technologies should be performed in a manner that allows reproduction in subsequent investigations and supports scientific advancement.

An automatic markerless registration method for neurosurgical robotics based on an optical camera.

PubMed

Meng, Fanle; Zhai, Fangwen; Zeng, Bowei; Ding, Hui; Wang, Guangzhi

2018-02-01

Current markerless registration methods for neurosurgical robotics use the facial surface to match the robot space with the image space, and acquisition of the facial surface usually requires manual interaction and constrains the patient to a supine position. To overcome these drawbacks, we propose a registration method that is automatic and does not constrain patient position. An optical camera attached to the robot end effector captures images around the patient's head from multiple views. Then, high coverage of the head surface is reconstructed from the images through multi-view stereo vision. Since the acquired head surface point cloud contains color information, a specific mark that is manually drawn on the patient's head prior to the capture procedure can be extracted to automatically accomplish coarse registration rather than using facial anatomic landmarks. Then, fine registration is achieved by registering the high coverage of the head surface without relying solely on the facial region, thus eliminating patient position constraints. The head surface was acquired by the camera with a good repeatability accuracy. The average target registration error of 8 different patient positions measured with targets inside a head phantom was [Formula: see text], while the mean surface registration error was [Formula: see text]. The method proposed in this paper achieves automatic markerless registration in multiple patient positions and guarantees registration accuracy inside the head. This method provides a new approach for establishing the spatial relationship between the image space and the robot space.

Improved accuracy of markerless motion tracking on bone suppression images: preliminary study for image-guided radiation therapy (IGRT)

NASA Astrophysics Data System (ADS)

Tanaka, Rie; Sanada, Shigeru; Sakuta, Keita; Kawashima, Hiroki

2015-05-01

The bone suppression technique based on advanced image processing can suppress the conspicuity of bones on chest radiographs, creating soft tissue images obtained by the dual-energy subtraction technique. This study was performed to evaluate the usefulness of bone suppression image processing in image-guided radiation therapy. We demonstrated the improved accuracy of markerless motion tracking on bone suppression images. Chest fluoroscopic images of nine patients with lung nodules during respiration were obtained using a flat-panel detector system (120 kV, 0.1 mAs/pulse, 5 fps). Commercial bone suppression image processing software was applied to the fluoroscopic images to create corresponding bone suppression images. Regions of interest were manually located on lung nodules and automatic target tracking was conducted based on the template matching technique. To evaluate the accuracy of target tracking, the maximum tracking error in the resulting images was compared with that of conventional fluoroscopic images. The tracking errors were decreased by half in eight of nine cases. The average maximum tracking errors in bone suppression and conventional fluoroscopic images were 1.3   ±   1.0 and 3.3   ±   3.3 mm, respectively. The bone suppression technique was especially effective in the lower lung area where pulmonary vessels, bronchi, and ribs showed complex movements. The bone suppression technique improved tracking accuracy without special equipment and implantation of fiducial markers, and with only additional small dose to the patient. Bone suppression fluoroscopy is a potential measure for respiratory displacement of the target. This paper was presented at RSNA 2013 and was carried out at Kanazawa University, JAPAN.

FlyCap: Markerless Motion Capture Using Multiple Autonomous Flying Cameras.

PubMed

Xu, Lan; Liu, Yebin; Cheng, Wei; Guo, Kaiwen; Zhou, Guyue; Dai, Qionghai; Fang, Lu

2017-07-18

Aiming at automatic, convenient and non-instrusive motion capture, this paper presents a new generation markerless motion capture technique, the FlyCap system, to capture surface motions of moving characters using multiple autonomous flying cameras (autonomous unmanned aerial vehicles(UAVs) each integrated with an RGBD video camera). During data capture, three cooperative flying cameras automatically track and follow the moving target who performs large-scale motions in a wide space. We propose a novel non-rigid surface registration method to track and fuse the depth of the three flying cameras for surface motion tracking of the moving target, and simultaneously calculate the pose of each flying camera. We leverage the using of visual-odometry information provided by the UAV platform, and formulate the surface tracking problem in a non-linear objective function that can be linearized and effectively minimized through a Gaussian-Newton method. Quantitative and qualitative experimental results demonstrate the plausible surface and motion reconstruction results.

Markerless video analysis for movement quantification in pediatric epilepsy monitoring.

PubMed

Lu, Haiping; Eng, How-Lung; Mandal, Bappaditya; Chan, Derrick W S; Ng, Yen-Ling

2011-01-01

This paper proposes a markerless video analytic system for quantifying body part movements in pediatric epilepsy monitoring. The system utilizes colored pajamas worn by a patient in bed to extract body part movement trajectories, from which various features can be obtained for seizure detection and analysis. Hence, it is non-intrusive and it requires no sensor/marker to be attached to the patient's body. It takes raw video sequences as input and a simple user-initialization indicates the body parts to be examined. In background/foreground modeling, Gaussian mixture models are employed in conjunction with HSV-based modeling. Body part detection follows a coarse-to-fine paradigm with graph-cut-based segmentation. Finally, body part parameters are estimated with domain knowledge guidance. Experimental studies are reported on sequences captured in an Epilepsy Monitoring Unit at a local hospital. The results demonstrate the feasibility of the proposed system in pediatric epilepsy monitoring and seizure detection.

Multithreaded hybrid feature tracking for markerless augmented reality.

PubMed

Lee, Taehee; Höllerer, Tobias

2009-01-01

We describe a novel markerless camera tracking approach and user interaction methodology for augmented reality (AR) on unprepared tabletop environments. We propose a real-time system architecture that combines two types of feature tracking. Distinctive image features of the scene are detected and tracked frame-to-frame by computing optical flow. In order to achieve real-time performance, multiple operations are processed in a synchronized multi-threaded manner: capturing a video frame, tracking features using optical flow, detecting distinctive invariant features, and rendering an output frame. We also introduce user interaction methodology for establishing a global coordinate system and for placing virtual objects in the AR environment by tracking a user's outstretched hand and estimating a camera pose relative to it. We evaluate the speed and accuracy of our hybrid feature tracking approach, and demonstrate a proof-of-concept application for enabling AR in unprepared tabletop environments, using bare hands for interaction.

Development of a morphology-based modeling technique for tracking solid-body displacements: examining the reliability of a potential MRI-only approach for joint kinematics assessment.

PubMed

Mahato, Niladri K; Montuelle, Stephane; Cotton, John; Williams, Susan; Thomas, James; Clark, Brian

2016-05-18

Single or biplanar video radiography and Roentgen stereophotogrammetry (RSA) techniques used for the assessment of in-vivo joint kinematics involves application of ionizing radiation, which is a limitation for clinical research involving human subjects. To overcome this limitation, our long-term goal is to develop a magnetic resonance imaging (MRI)-only, three dimensional (3-D) modeling technique that permits dynamic imaging of joint motion in humans. Here, we present our initial findings, as well as reliability data, for an MRI-only protocol and modeling technique. We developed a morphology-based motion-analysis technique that uses MRI of custom-built solid-body objects to animate and quantify experimental displacements between them. The technique involved four major steps. First, the imaging volume was calibrated using a custom-built grid. Second, 3-D models were segmented from axial scans of two custom-built solid-body cubes. Third, these cubes were positioned at pre-determined relative displacements (translation and rotation) in the magnetic resonance coil and scanned with a T1 and a fast contrast-enhanced pulse sequences. The digital imaging and communications in medicine (DICOM) images were then processed for animation. The fourth step involved importing these processed images into an animation software, where they were displayed as background scenes. In the same step, 3-D models of the cubes were imported into the animation software, where the user manipulated the models to match their outlines in the scene (rotoscoping) and registered the models into an anatomical joint system. Measurements of displacements obtained from two different rotoscoping sessions were tested for reliability using coefficient of variations (CV), intraclass correlation coefficients (ICC), Bland-Altman plots, and Limits of Agreement analyses. Between-session reliability was high for both the T1 and the contrast-enhanced sequences. Specifically, the average CVs for translation were 4.31 % and 5.26 % for the two pulse sequences, respectively, while the ICCs were 0.99 for both. For rotation measures, the CVs were 3.19 % and 2.44 % for the two pulse sequences with the ICCs being 0.98 and 0.97, respectively. A novel biplanar imaging approach also yielded high reliability with mean CVs of 2.66 % and 3.39 % for translation in the x- and z-planes, respectively, and ICCs of 0.97 in both planes. This work provides basic proof-of-concept for a reliable marker-less non-ionizing-radiation-based quasi-dynamic motion quantification technique that can potentially be developed into a tool for real-time joint kinematics analysis.

Genome-wide Screening Identifies Phosphotransferase System Permease BepA to Be Involved in Enterococcus faecium Endocarditis and Biofilm Formation.

PubMed

Paganelli, Fernanda L; Huebner, Johannes; Singh, Kavindra V; Zhang, Xinglin; van Schaik, Willem; Wobser, Dominique; Braat, Johanna C; Murray, Barbara E; Bonten, Marc J M; Willems, Rob J L; Leavis, Helen L

2016-07-15

Enterococcus faecium is a common cause of nosocomial infections, of which infective endocarditis is associated with substantial mortality. In this study, we used a microarray-based transposon mapping (M-TraM) approach to evaluate a rat endocarditis model and identified a gene, originally annotated as "fruA" and renamed "bepA," putatively encoding a carbohydrate phosphotransferase system (PTS) permease (biofilm and endocarditis-associated permease A [BepA]), as important in infective endocarditis. This gene is highly enriched in E. faecium clinical isolates and absent in commensal isolates that are not associated with infection. Confirmation of the phenotype was established in a competition experiment of wild-type and a markerless bepA mutant in a rat endocarditis model. In addition, deletion of bepA impaired biofilm formation in vitro in the presence of 100% human serum and metabolism of β-methyl-D-glucoside. β-glucoside metabolism has been linked to the metabolism of glycosaminoglycans that are exposed on injured heart valves, where bacteria attach and form vegetations. Therefore, we propose that the PTS permease BepA is directly implicated in E. faecium pathogenesis. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers.

PubMed

Weninger, Astrid; Fischer, Jasmin E; Raschmanová, Hana; Kniely, Claudia; Vogl, Thomas; Glieder, Anton

2018-04-01

Komagataella phaffii (syn. Pichia pastoris) is one of the most commonly used host systems for recombinant protein expression. Achieving targeted genetic modifications had been hindered by low frequencies of homologous recombination (HR). Recently, a CRISPR/Cas9 genome editing system has been implemented for P. pastoris enabling gene knockouts based on indels (insertion, deletions) via non-homologous end joining (NHEJ) at near 100% efficiency. However, specifically integrating homologous donor cassettes via HR for replacement studies had proven difficult resulting at most in ∼20% correct integration using CRISPR/Cas9. Here, we demonstrate the CRISPR/Cas9 mediated integration of markerless donor cassettes at efficiencies approaching 100% using a ku70 deletion strain. The Ku70p is involved in NHEJ repair and lack of the protein appears to favor repair via HR near exclusively. While the absolute number of transformants in the Δku70 strain is reduced, virtually all surviving transformants showed correct integration. In the wildtype strain, markerless donor cassette integration was also improved up to 25-fold by placing an autonomously replicating sequence (ARS) on the donor cassette. Alternative strategies for improving donor cassette integration using a Cas9 nickase variant or reducing off targeting associated toxicity using a high fidelity Cas9 variant were so far not successful in our hands in P. pastoris. Furthermore we provide Cas9/gRNA expression plasmids with a Geneticin resistance marker which proved to be versatile tools for marker recycling. The reported CRSIPR-Cas9 tools can be applied for modifying existing production strains and also pave the way for markerless whole genome modification studies in P. pastoris. © 2017 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc.

Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers

PubMed Central

Weninger, Astrid; Fischer, Jasmin E.; Raschmanová, Hana; Kniely, Claudia; Glieder, Anton

2017-01-01

Abstract Komagataella phaffii (syn. Pichia pastoris) is one of the most commonly used host systems for recombinant protein expression. Achieving targeted genetic modifications had been hindered by low frequencies of homologous recombination (HR). Recently, a CRISPR/Cas9 genome editing system has been implemented for P. pastoris enabling gene knockouts based on indels (insertion, deletions) via non‐homologous end joining (NHEJ) at near 100% efficiency. However, specifically integrating homologous donor cassettes via HR for replacement studies had proven difficult resulting at most in ∼20% correct integration using CRISPR/Cas9. Here, we demonstrate the CRISPR/Cas9 mediated integration of markerless donor cassettes at efficiencies approaching 100% using a ku70 deletion strain. The Ku70p is involved in NHEJ repair and lack of the protein appears to favor repair via HR near exclusively. While the absolute number of transformants in the Δku70 strain is reduced, virtually all surviving transformants showed correct integration. In the wildtype strain, markerless donor cassette integration was also improved up to 25‐fold by placing an autonomously replicating sequence (ARS) on the donor cassette. Alternative strategies for improving donor cassette integration using a Cas9 nickase variant or reducing off targeting associated toxicity using a high fidelity Cas9 variant were so far not successful in our hands in P. pastoris. Furthermore we provide Cas9/gRNA expression plasmids with a Geneticin resistance marker which proved to be versatile tools for marker recycling. The reported CRSIPR‐Cas9 tools can be applied for modifying existing production strains and also pave the way for markerless whole genome modification studies in P. pastoris. PMID:29091307

Automatic markerless registration of point clouds with semantic-keypoint-based 4-points congruent sets

NASA Astrophysics Data System (ADS)

Ge, Xuming

2017-08-01

The coarse registration of point clouds from urban building scenes has become a key topic in applications of terrestrial laser scanning technology. Sampling-based algorithms in the random sample consensus (RANSAC) model have emerged as mainstream solutions to address coarse registration problems. In this paper, we propose a novel combined solution to automatically align two markerless point clouds from building scenes. Firstly, the method segments non-ground points from ground points. Secondly, the proposed method detects feature points from each cross section and then obtains semantic keypoints by connecting feature points with specific rules. Finally, the detected semantic keypoints from two point clouds act as inputs to a modified 4PCS algorithm. Examples are presented and the results compared with those of K-4PCS to demonstrate the main contributions of the proposed method, which are the extension of the original 4PCS to handle heavy datasets and the use of semantic keypoints to improve K-4PCS in relation to registration accuracy and computational efficiency.

A natural user interface to integrate citizen science and physical exercise.

PubMed

Palermo, Eduardo; Laut, Jeffrey; Nov, Oded; Cappa, Paolo; Porfiri, Maurizio

2017-01-01

Citizen science enables volunteers to contribute to scientific projects, where massive data collection and analysis are often required. Volunteers participate in citizen science activities online from their homes or in the field and are motivated by both intrinsic and extrinsic factors. Here, we investigated the possibility of integrating citizen science tasks within physical exercises envisaged as part of a potential rehabilitation therapy session. The citizen science activity entailed environmental mapping of a polluted body of water using a miniature instrumented boat, which was remotely controlled by the participants through their physical gesture tracked by a low-cost markerless motion capture system. Our findings demonstrate that the natural user interface offers an engaging and effective means for performing environmental monitoring tasks. At the same time, the citizen science activity increases the commitment of the participants, leading to a better motion performance, quantified through an array of objective indices. The study constitutes a first and necessary step toward rehabilitative treatments of the upper limb through citizen science and low-cost markerless optical systems.

Fluoroscopic tumor tracking for image-guided lung cancer radiotherapy

NASA Astrophysics Data System (ADS)

Lin, Tong; Cerviño, Laura I.; Tang, Xiaoli; Vasconcelos, Nuno; Jiang, Steve B.

2009-02-01

Accurate lung tumor tracking in real time is a keystone to image-guided radiotherapy of lung cancers. Existing lung tumor tracking approaches can be roughly grouped into three categories: (1) deriving tumor position from external surrogates; (2) tracking implanted fiducial markers fluoroscopically or electromagnetically; (3) fluoroscopically tracking lung tumor without implanted fiducial markers. The first approach suffers from insufficient accuracy, while the second may not be widely accepted due to the risk of pneumothorax. Previous studies in fluoroscopic markerless tracking are mainly based on template matching methods, which may fail when the tumor boundary is unclear in fluoroscopic images. In this paper we propose a novel markerless tumor tracking algorithm, which employs the correlation between the tumor position and surrogate anatomic features in the image. The positions of the surrogate features are not directly tracked; instead, we use principal component analysis of regions of interest containing them to obtain parametric representations of their motion patterns. Then, the tumor position can be predicted from the parametric representations of surrogates through regression. Four regression methods were tested in this study: linear and two-degree polynomial regression, artificial neural network (ANN) and support vector machine (SVM). The experimental results based on fluoroscopic sequences of ten lung cancer patients demonstrate a mean tracking error of 2.1 pixels and a maximum error at a 95% confidence level of 4.6 pixels (pixel size is about 0.5 mm) for the proposed tracking algorithm.

Construction of new cloning, lacZ reporter and scarless-markerless suicide vectors for genetic studies in Aggregatibacter actinomycetemcomitans

PubMed Central

Juárez-Rodríguez, María Dolores; Torres-Escobar, Ascención; Demuth, Donald R.

2013-01-01

To elucidate the putative function of a gene, effective tools are required for genetic characterization that facilitate its inactivation, deletion or modification on the bacterial chromosome. In the present study, the nucleotide sequence of the Escherichia coli/Aggregatibacter actinomycetemcomitans shuttle vector pYGK was determined, allowing us to redesign and construct a new shuttle cloning vector, pJT4, and promoterless lacZ transcriptional/translational fusion plasmids, pJT3 and pJT5. Plasmids pJT4 and pJT5 contain the origin of replication necessary to maintain shuttle vector replication. In addition, a new suicide vector, pJT1, was constructed for the generation of scarless and markerless deletion mutations of genes in the oral pathogen A. actinomycetemcomitans. Plasmid pJT1 is a pUC-based suicide vector that is counter-selectable for sucrose sensitivity. This vector does not leave antibiotic markers or scars on the chromosome after gene deletion and thus provides the option to combine several mutations in the same genetic background. The effectiveness of pJT1 was demonstrated by the construction of A. actinomycetemcomitans isogenic qseB single deletion (ΔqseB) mutant and lsrRK double deletion mutants (ΔlsrRK). These new vectors may offer alternatives for genetic studies in A. actinomycetemcomitans and other members of the HACEK (Haemophilus spp., A. actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae) group of Gram-negative bacteria. PMID:23353051

DOE Office of Scientific and Technical Information (OSTI.GOV)

Panfil, J; Patel, R; Surucu, M

Purpose: To compare markerless template-based tracking of lung tumors using dual energy (DE) cone-beam computed tomography (CBCT) projections versus single energy (SE) CBCT projections. Methods: A RANDO chest phantom with a simulated tumor in the upper right lung was used to investigate the effectiveness of tumor tracking using DE and SE CBCT projections. Planar kV projections from CBCT acquisitions were captured at 60 kVp (4 mAs) and 120 kVp (1 mAs) using the Varian TrueBeam and non-commercial iTools Capture software. Projections were taken at approximately every 0.53° while the gantry rotated. Due to limitations of the phantom, angles for whichmore » the shoulders blocked the tumor were excluded from tracking analysis. DE images were constructed using a weighted logarithmic subtraction that removed bony anatomy while preserving soft tissue structures. The tumors were tracked separately on DE and SE (120 kVp) images using a template-based tracking algorithm. The tracking results were compared to ground truth coordinates designated by a physician. Matches with a distance of greater than 3 mm from ground truth were designated as failing to track. Results: 363 frames were analyzed. The algorithm successfully tracked the tumor on 89.8% (326/363) of DE frames compared to 54.3% (197/363) of SE frames (p<0.0001). Average distance between tracking and ground truth coordinates was 1.27 +/− 0.67 mm for DE versus 1.83+/−0.74 mm for SE (p<0.0001). Conclusion: This study demonstrates the effectiveness of markerless template-based tracking using DE CBCT. DE imaging resulted in better detectability with more accurate localization on average versus SE. Supported by a grant from Varian Medical Systems.« less

Biomechanical analysis of three tennis serve types using a markerless system.

PubMed

Abrams, Geoffrey D; Harris, Alex H S; Andriacchi, Thomas P; Safran, Marc R

2014-02-01

The tennis serve is commonly associated with musculoskeletal injury. Advanced players are able to hit multiple serve types with different types of spin. No investigation has characterised the kinematics of all three serve types for the upper extremity and back. Seven NCAA Division I male tennis players performed three successful flat, kick and slice serves. Serves were recorded using an eight camera markerless motion capture system. Laser scanning was utilised to accurately collect body dimensions and data were computed using inverse kinematic methods. There was no significant difference in maximum back extension angle for the flat, kick or slice serves. The kick serve had a higher force magnitude at the back than the flat and slice as well as larger posteriorly directed shoulder forces. The flat serve had significantly greater maximum shoulder internal rotation velocity versus the slice serve. Force and torque magnitudes at the elbow and wrist were not significantly different between the serves. The kick serve places higher physical demands on the back and shoulder while the slice serve demonstrated lower overall kinetic forces. This information may have injury prevention and rehabilitation implications.

A natural user interface to integrate citizen science and physical exercise

PubMed Central

Palermo, Eduardo; Laut, Jeffrey; Nov, Oded; Porfiri, Maurizio

2017-01-01

Citizen science enables volunteers to contribute to scientific projects, where massive data collection and analysis are often required. Volunteers participate in citizen science activities online from their homes or in the field and are motivated by both intrinsic and extrinsic factors. Here, we investigated the possibility of integrating citizen science tasks within physical exercises envisaged as part of a potential rehabilitation therapy session. The citizen science activity entailed environmental mapping of a polluted body of water using a miniature instrumented boat, which was remotely controlled by the participants through their physical gesture tracked by a low-cost markerless motion capture system. Our findings demonstrate that the natural user interface offers an engaging and effective means for performing environmental monitoring tasks. At the same time, the citizen science activity increases the commitment of the participants, leading to a better motion performance, quantified through an array of objective indices. The study constitutes a first and necessary step toward rehabilitative treatments of the upper limb through citizen science and low-cost markerless optical systems. PMID:28231261

Markerless motion estimation for motion-compensated clinical brain imaging

NASA Astrophysics Data System (ADS)

Kyme, Andre Z.; Se, Stephen; Meikle, Steven R.; Fulton, Roger R.

2018-05-01

Motion-compensated brain imaging can dramatically reduce the artifacts and quantitative degradation associated with voluntary and involuntary subject head motion during positron emission tomography (PET), single photon emission computed tomography (SPECT) and computed tomography (CT). However, motion-compensated imaging protocols are not in widespread clinical use for these modalities. A key reason for this seems to be the lack of a practical motion tracking technology that allows for smooth and reliable integration of motion-compensated imaging protocols in the clinical setting. We seek to address this problem by investigating the feasibility of a highly versatile optical motion tracking method for PET, SPECT and CT geometries. The method requires no attached markers, relying exclusively on the detection and matching of distinctive facial features. We studied the accuracy of this method in 16 volunteers in a mock imaging scenario by comparing the estimated motion with an accurate marker-based method used in applications such as image guided surgery. A range of techniques to optimize performance of the method were also studied. Our results show that the markerless motion tracking method is highly accurate (<2 mm discrepancy against a benchmarking system) on an ethnically diverse range of subjects and, moreover, exhibits lower jitter and estimation of motion over a greater range than some marker-based methods. Our optimization tests indicate that the basic pose estimation algorithm is very robust but generally benefits from rudimentary background masking. Further marginal gains in accuracy can be achieved by accounting for non-rigid motion of features. Efficiency gains can be achieved by capping the number of features used for pose estimation provided that these features adequately sample the range of head motion encountered in the study. These proof-of-principle data suggest that markerless motion tracking is amenable to motion-compensated brain imaging and holds good promise for a practical implementation in clinical PET, SPECT and CT systems.

NOTE: A feasibility study of markerless fluoroscopic gating for lung cancer radiotherapy using 4DCT templates

NASA Astrophysics Data System (ADS)

Li, Ruijiang; Lewis, John H.; Cerviño, Laura I.; Jiang, Steve B.

2009-10-01

A major difficulty in conformal lung cancer radiotherapy is respiratory organ motion, which may cause clinically significant targeting errors. Respiratory-gated radiotherapy allows for more precise delivery of prescribed radiation dose to the tumor, while minimizing normal tissue complications. Gating based on external surrogates is limited by its lack of accuracy, while gating based on implanted fiducial markers is limited primarily by the risk of pneumothorax due to marker implantation. Techniques for fluoroscopic gating without implanted fiducial markers (markerless gating) have been developed. These techniques usually require a training fluoroscopic image dataset with marked tumor positions in the images, which limits their clinical implementation. To remove this requirement, this study presents a markerless fluoroscopic gating algorithm based on 4DCT templates. To generate gating signals, we explored the application of three similarity measures or scores between fluoroscopic images and the reference 4DCT template: un-normalized cross-correlation (CC), normalized cross-correlation (NCC) and normalized mutual information (NMI), as well as average intensity (AI) of the region of interest (ROI) in the fluoroscopic images. Performance was evaluated using fluoroscopic and 4DCT data from three lung cancer patients. On average, gating based on CC achieves the highest treatment accuracy given the same efficiency, with a high target coverage (average between 91.9% and 98.6%) for a wide range of nominal duty cycles (20-50%). AI works well for two patients out of three, but failed for the third patient due to interference from the heart. Gating based on NCC and NMI usually failed below 50% nominal duty cycle. Based on this preliminary study with three patients, we found that the proposed CC-based gating algorithm can generate accurate and robust gating signals when using 4DCT reference template. However, this observation is based on results obtained from a very limited dataset, and further investigation on a larger patient population has to be done before its clinical implementation.

SU-D-207-01: Markerless Respiratory Motion Tracking with Contrast Enhanced Thoracic Cone Beam CT Projections

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chao, M; Yuan, Y; Rosenzweig, K

2015-06-15

Purpose: To develop a novel technique to enhance the image contrast of clinical cone beam CT projections and extract respiratory signals based on anatomical motion using the modified Amsterdam Shroud (AS) method to benefit image guided radiation therapy. Methods: Thoracic cone beam CT projections acquired prior to treatment were preprocessed to increase their contrast for better respiratory signal extraction. Air intensity on raw images was firstly estimated and then applied to correct the projections to generate new attenuation images that were subsequently improved with deeper anatomy feature enhancement through taking logarithm operation, derivative along superior-inferior direction, respectively. All pixels onmore » individual post-processed two dimensional images were horizontally summed to one column and all projections were combined side by side to create an AS image from which patient’s respiratory signal was extracted. The impact of gantry rotation on the breathing signal rendering was also investigated. Ten projection image sets from five lung cancer patients acquired with the Varian Onboard Imager on 21iX Clinac (Varian Medical Systems, Palo Alto, CA) were employed to assess the proposed technique. Results: Application of the air correction on raw projections showed that more than an order of magnitude of contrast enhancement was achievable. The typical contrast on the raw projections is around 0.02 while that on attenuation images could greater than 0.5. Clear and stable breathing signal can be reliably extracted from the new images while the uncorrected projection sets failed to yield clear signals most of the time. Conclusion: Anatomy feature plays a key role in yielding breathing signal from the projection images using the AS technique. The air correction process facilitated the contrast enhancement significantly and attenuation images thus obtained provides a practical solution to obtaining markerless breathing motion tracking.« less

[Construction of Corynebacterium crenatum AS 1.542 δ argR and analysis of transcriptional levels of the related genes of arginine biosynthetic pathway].

PubMed

Chen, Xuelan; Tang, Li; Jiao, Haitao; Xu, Feng; Xiong, Yonghua

2013-01-04

ArgR, coded by the argR gene from Corynebacterium crenatum AS 1.542, acts as a negative regulator in arginine biosynthetic pathway. However, the effect of argR on transcriptional levels of the related biosynthetic genes has not been reported. Here, we constructed a deletion mutant of argR gene: C. crenatum AS 1.542 Delta argR using marker-less knockout technology, and compared the changes of transcriptional levels of the arginine biosynthetic genes between the mutant strain and the wild-type strain. We used marker-less knockout technology to construct C. crenatum AS 1.542 Delta argR and analyzed the changes of the relate genes at the transcriptional level using real-time fluorescence quantitative PCR. C. crenatum AS 1.542 Delta argR was successfully obtained and the transcriptional level of arginine biosynthetic genes in this mutant increased significantly with an average of about 162.1 folds. The arginine biosynthetic genes in C. crenatum are clearly controlled by the negative regulator ArgR. However, the deletion of this regulator does not result in a clear change in arginine production in the bacteria.

A Marker-less Monitoring System for Movement Analysis of Infants Using Video Images

NASA Astrophysics Data System (ADS)

Shima, Keisuke; Osawa, Yuko; Bu, Nan; Tsuji, Tokuo; Tsuji, Toshio; Ishii, Idaku; Matsuda, Hiroshi; Orito, Kensuke; Ikeda, Tomoaki; Noda, Shunichi

This paper proposes a marker-less motion measurement and analysis system for infants. This system calculates eight types of evaluation indices related to the movement of an infant such as “amount of body motion” and “activity of body” from binary images that are extracted from video images using the background difference and frame difference. Thus, medical doctors can intuitively understand the movements of infants without long-term observations, and this may be helpful in supporting their diagnoses and detecting disabilities and diseases in the early stages. The distinctive feature of this system is that the movements of infants can be measured without using any markers for motion capture and thus it is expected that the natural and inherent tendencies of infants can be analyzed and evaluated. In this paper, the evaluation indices and features of movements between full-term infants (FTIs) and low birth weight infants (LBWIs) are compared using the developed prototype. We found that the amount of body motion and symmetry of upper and lower body movements of LBWIs became lower than those of FTIs. The difference between the movements of FTIs and LBWIs can be evaluated using the proposed system.

SU-G-JeP1-11: Feasibility Study of Markerless Tracking Using Dual Energy Fluoroscopic Images for Real-Time Tumor-Tracking Radiotherapy System

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shiinoki, T; Shibuya, K; Sawada, A

Purpose: The new real-time tumor-tracking radiotherapy (RTRT) system was installed in our institution. This system consists of two x-ray tubes and color image intensifiers (I.I.s). The fiducial marker which was implanted near the tumor was tracked using color fluoroscopic images. However, the implantation of the fiducial marker is very invasive. Color fluoroscopic images enable to increase the recognition of the tumor. However, these images were not suitable to track the tumor without fiducial marker. The purpose of this study was to investigate the feasibility of markerless tracking using dual energy colored fluoroscopic images for real-time tumor-tracking radiotherapy system. Methods: Themore » colored fluoroscopic images of static and moving phantom that had the simulated tumor (30 mm diameter sphere) were experimentally acquired using the RTRT system. The programmable respiratory motion phantom was driven using the sinusoidal pattern in cranio-caudal direction (Amplitude: 20 mm, Time: 4 s). The x-ray condition was set to 55 kV, 50 mA and 105 kV, 50 mA for low energy and high energy, respectively. Dual energy images were calculated based on the weighted logarithmic subtraction of high and low energy images of RGB images. The usefulness of dual energy imaging for real-time tracking with an automated template image matching algorithm was investigated. Results: Our proposed dual energy subtraction improve the contrast between tumor and background to suppress the bone structure. For static phantom, our results showed that high tracking accuracy using dual energy subtraction images. For moving phantom, our results showed that good tracking accuracy using dual energy subtraction images. However, tracking accuracy was dependent on tumor position, tumor size and x-ray conditions. Conclusion: We indicated that feasibility of markerless tracking using dual energy fluoroscopic images for real-time tumor-tracking radiotherapy system. Furthermore, it is needed to investigate the tracking accuracy using proposed dual energy subtraction images for clinical cases.« less

Carbon-Ion Pencil Beam Scanning Treatment With Gated Markerless Tumor Tracking: An Analysis of Positional Accuracy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mori, Shinichiro, E-mail: shinshin@nirs.go.jp; Karube, Masataka; Shirai, Toshiyuki

Purpose: Having implemented amplitude-based respiratory gating for scanned carbon-ion beam therapy, we sought to evaluate its effect on positional accuracy and throughput. Methods and Materials: A total of 10 patients with tumors of the lung and liver participated in the first clinical trials at our center. Treatment planning was conducted with 4-dimensional computed tomography (4DCT) under free-breathing conditions. The planning target volume (PTV) was calculated by adding a 2- to 3-mm setup margin outside the clinical target volume (CTV) within the gating window. The treatment beam was on when the CTV was within the PTV. Tumor position was detected inmore » real time with a markerless tumor tracking system using paired x-ray fluoroscopic imaging units. Results: The patient setup error (mean ± SD) was 1.1 ± 1.2 mm/0.6 ± 0.4°. The mean internal gating accuracy (95% confidence interval [CI]) was 0.5 mm. If external gating had been applied to this treatment, the mean gating accuracy (95% CI) would have been 4.1 mm. The fluoroscopic radiation doses (mean ± SD) were 23.7 ± 21.8 mGy per beam and less than 487.5 mGy total throughout the treatment course. The setup, preparation, and irradiation times (mean ± SD) were 8.9 ± 8.2 min, 9.5 ± 4.6 min, and 4.0 ± 2.4 min, respectively. The treatment room occupation time was 36.7 ± 67.5 min. Conclusions: Internal gating had a much higher accuracy than external gating. By the addition of a setup margin of 2 to 3 mm, internal gating positional error was less than 2.2 mm at 95% CI.« less

Markerless EPID image guided dynamic multi-leaf collimator tracking for lung tumors

NASA Astrophysics Data System (ADS)

Rottmann, J.; Keall, P.; Berbeco, R.

2013-06-01

Compensation of target motion during the delivery of radiotherapy has the potential to improve treatment accuracy, dose conformity and sparing of healthy tissue. We implement an online image guided therapy system based on soft tissue localization (STiL) of the target from electronic portal images and treatment aperture adaptation with a dynamic multi-leaf collimator (DMLC). The treatment aperture is moved synchronously and in real time with the tumor during the entire breathing cycle. The system is implemented and tested on a Varian TX clinical linear accelerator featuring an AS-1000 electronic portal imaging device (EPID) acquiring images at a frame rate of 12.86 Hz throughout the treatment. A position update cycle for the treatment aperture consists of four steps: in the first step at time t = t0 a frame is grabbed, in the second step the frame is processed with the STiL algorithm to get the tumor position at t = t0, in a third step the tumor position at t = ti + δt is predicted to overcome system latencies and in the fourth step, the DMLC control software calculates the required leaf motions and applies them at time t = ti + δt. The prediction model is trained before the start of the treatment with data representing the tumor motion. We analyze the system latency with a dynamic chest phantom (4D motion phantom, Washington University). We estimate the average planar position deviation between target and treatment aperture in a clinical setting by driving the phantom with several lung tumor trajectories (recorded from fiducial tracking during radiotherapy delivery to the lung). DMLC tracking for lung stereotactic body radiation therapy without fiducial markers was successfully demonstrated. The inherent system latency is found to be δt = (230 ± 11) ms for a MV portal image acquisition frame rate of 12.86 Hz. The root mean square deviation between tumor and aperture position is smaller than 1 mm. We demonstrate the feasibility of real-time markerless DMLC tracking with a standard LINAC-mounted (EPID).

See It With Your Own Eyes: Markerless Mobile Augmented Reality for Radiation Awareness in the Hybrid Room.

PubMed

Loy Rodas, Nicolas; Barrera, Fernando; Padoy, Nicolas

2017-02-01

We present an approach to provide awareness to the harmful ionizing radiation generated during X-ray-guided minimally invasive procedures. A hand-held screen is used to display directly in the user's view information related to radiation safety in a mobile augmented reality (AR) manner. Instead of using markers, we propose a method to track the observer's viewpoint, which relies on the use of multiple RGB-D sensors and combines equipment detection for tracking initialization with a KinectFusion-like approach for frame-to-frame tracking. Two of the sensors are ceiling-mounted and a third one is attached to the hand-held screen. The ceiling cameras keep an updated model of the room's layout, which is used to exploit context information and improve the relocalization procedure. The system is evaluated on a multicamera dataset generated inside an operating room (OR) and containing ground-truth poses of the AR display. This dataset includes a wide variety of sequences with different scene configurations, occlusions, motion in the scene, and abrupt viewpoint changes. Qualitative results illustrating the different AR visualization modes for radiation awareness provided by the system are also presented. Our approach allows the user to benefit from a large AR visualization area and permits to recover from tracking failure caused by vast motion or changes in the scene just by looking at a piece of equipment. The system enables the user to see the 3-D propagation of radiation, the medical staff's exposure, and/or the doses deposited on the patient's surface as seen through his own eyes.

Marker-less multi-frame motion tracking and compensation in PET-brain imaging

NASA Astrophysics Data System (ADS)

Lindsay, C.; Mukherjee, J. M.; Johnson, K.; Olivier, P.; Song, X.; Shao, L.; King, M. A.

2015-03-01

In PET brain imaging, patient motion can contribute significantly to the degradation of image quality potentially leading to diagnostic and therapeutic problems. To mitigate the image artifacts resulting from patient motion, motion must be detected and tracked then provided to a motion correction algorithm. Existing techniques to track patient motion fall into one of two categories: 1) image-derived approaches and 2) external motion tracking (EMT). Typical EMT requires patients to have markers in a known pattern on a rigid too attached to their head, which are then tracked by expensive and bulky motion tracking camera systems or stereo cameras. This has made marker-based EMT unattractive for routine clinical application. Our main contributions are the development of a marker-less motion tracking system that uses lowcost, small depth-sensing cameras which can be installed in the bore of the imaging system. Our motion tracking system does not require anything to be attached to the patient and can track the rigid transformation (6-degrees of freedom) of the patient's head at a rate 60 Hz. We show that our method can not only be used in with Multi-frame Acquisition (MAF) PET motion correction, but precise timing can be employed to determine only the necessary frames needed for correction. This can speeds up reconstruction by eliminating the unnecessary subdivision of frames.

The validity and intra-tester reliability of markerless motion capture to analyse kinematics of the BMX Supercross gate start.

PubMed

Grigg, Josephine; Haakonssen, Eric; Rathbone, Evelyne; Orr, Robin; Keogh, Justin W L

2017-11-13

The aim of this study was to quantify the validity and intra-tester reliability of a novel method of kinematic measurement. The measurement target was the joint angles of an athlete performing a BMX Supercross (SX) gate start action through the first 1.2 s of movement in situ on a BMX SX ramp using a standard gate start procedure. The method employed GoPro® Hero 4 Silver (GoPro Inc., USA) cameras capturing data at 120 fps 720 p on a 'normal' lens setting. Kinovea 0.8.15 (Kinovea.org, France) was used for analysis. Tracking data was exported and angles computed in Matlab (Mathworks®, USA). The gold standard 3D method for joint angle measurement could not safely be employed in this environment, so a rigid angle was used. Validity was measured to be within 2°. Intra-tester reliability was measured by the same tester performing the analysis twice with an average of 55 days between analyses. Intra-tester reliability was high, with an absolute error <6° and <9 frames (0.075 s) across all angles and time points for key positions, respectively. The methodology is valid within 2° and reliable within 6° for the calculation of joint angles in the first ~1.25 s.

A study of sea lion hydrodynamics using a robotic foreflipper platform

NASA Astrophysics Data System (ADS)

Kulkarni, Aditya A.; Patel, Rahi K.; Leftwich, Megan C.

2016-11-01

Unlike most fish and mammals-that utilize BCF swimming-sea lions rely on their foreflippers to generate thrust without a characteristic flapping frequency. This unique swimming style allows the sea lion to be highly maneuverable, while also producing high amounts of thrust. To explore this motion, and the physics that underlies it, we use novel markerless tracking techniques on untrained sea lions at the Smithsonian National Zoo in Washington, D.C to get the complete motion during different maneuvers. High speed video and three-dimensional surface reconstruction techniques are used to extract the foreflippers kinematics during the thrust phase. Using this data, pitch angle is calculated with respect to the base of the flipper to build a scaled robotic flipper. Dye visualization is carried out in a water channel by injecting dye upstream of the leading edge of the flipper with flow speed set to explore different parameters, like Reynolds number or angular velocity. Results show low pressure on the upper surface of the flipper causes the fluid to be pulled around the flipper forming a vortex that moves fully out of the plane.

Comparison of two different Radiostereometric analysis (RSA) systems with markerless elementary geometrical shape modeling for the measurement of stem migration.

PubMed

Li, Ye; Röhrl, Stephan M; Bøe, B; Nordsletten, Lars

2014-09-01

Radiostereometric analysis (RSA) is the gold standard of measurement for in vivo 3D implants migration. The aim of this study was to evaluate the in vivo precision of 2 RSA marker-based systems compared with that of marker-free, elementary geometrical shape modeling RSA. Stem migration was measured in 50 patients recruited from an on-going Randomized Controlled Trial. We performed marker-based analysis with the Um RSA and RSAcore systems and compared these results with those of the elementary geometrical shape RSA. The precision for subsidence was 0.118 mm for Um RSA, 0.141 mm for RSAcore, and 0.136 mm for elementary geometrical shape RSA. The precision for retroversion was 1.3° for elementary geometrical shape RSA, approximately 2-fold greater than that for the other methods. The intraclass correlation coefficient between the marker-based systems and elementary geometrical shape RSA was approximately 0.5 for retroversion. All 3 methods yielded ICCs for subsidence and varus-valgus rotation above 0.9. We found an excellent correlation between marker-based RSA and elementary geometrical shape RSA for subsidence and varus-valgus rotation, independent of the system used. The precisions for out-of-plane migration were inferior for elementary geometrical shape RSA. Therefore, as a mechanism of failure, retroversion may be more difficult to detect early. This is to our knowledge the first study to compare different RSA systems with or without markers on the implant. Marker-based RSA has high precision in all planes, independent of the system used. Elementary geometrical shape RSA is inferior in out-of-plane migration. Copyright © 2014 Elsevier Ltd. All rights reserved.

Mobile markerless augmented reality and its application in forensic medicine.

PubMed

Kilgus, Thomas; Heim, Eric; Haase, Sven; Prüfer, Sabine; Müller, Michael; Seitel, Alexander; Fangerau, Markus; Wiebe, Tamara; Iszatt, Justin; Schlemmer, Heinz-Peter; Hornegger, Joachim; Yen, Kathrin; Maier-Hein, Lena

2015-05-01

During autopsy, forensic pathologists today mostly rely on visible indication, tactile perception and experience to determine the cause of death. Although computed tomography (CT) data is often available for the bodies under examination, these data are rarely used due to the lack of radiological workstations in the pathological suite. The data may prevent the forensic pathologist from damaging evidence by allowing him to associate, for example, external wounds to internal injuries. To facilitate this, we propose a new multimodal approach for intuitive visualization of forensic data and evaluate its feasibility. A range camera is mounted on a tablet computer and positioned in a way such that the camera simultaneously captures depth and color information of the body. A server estimates the camera pose based on surface registration of CT and depth data to allow for augmented reality visualization of the internal anatomy directly on the tablet. Additionally, projection of color information onto the CT surface is implemented. We validated the system in a postmortem pilot study using fiducials attached to the skin for quantification of a mean target registration error of [Formula: see text] mm. The system is mobile, markerless, intuitive and real-time capable with sufficient accuracy. It can support the forensic pathologist during autopsy with augmented reality and textured surfaces. Furthermore, the system enables multimodal documentation for presentation in court. Despite its preliminary prototype status, it has high potential due to its low price and simplicity.

Development of a Markerless Knockout Method for Actinobacillus succinogenes

PubMed Central

Joshi, Rajasi V.; Schindler, Bryan D.; McPherson, Nikolas R.; Tiwari, Kanupriya

2014-01-01

Actinobacillus succinogenes is one of the best natural succinate-producing organisms, but it still needs engineering to further increase succinate yield and productivity. In this study, we developed a markerless knockout method for A. succinogenes using natural transformation or electroporation. The Escherichia coli isocitrate dehydrogenase gene with flanking flippase recognition target sites was used as the positive selection marker, making use of A. succinogenes's auxotrophy for glutamate to select for growth on isocitrate. The Saccharomyces cerevisiae flippase recombinase (Flp) was used to remove the selection marker, allowing its reuse. Finally, the plasmid expressing flp was cured using acridine orange. We demonstrate that at least two consecutive deletions can be introduced into the same strain using this approach, that no more than a total of 1 kb of DNA is needed on each side of the selection cassette to protect from exonuclease activity during transformation, and that no more than 200 bp of homologous DNA is needed on each side for efficient recombination. We also demonstrate that electroporation can be used as an alternative transformation method to obtain knockout mutants and that an enriched defined medium can be used for direct selection of knockout mutants on agar plates with high efficiency. Single-knockout mutants of the fumarate reductase and of the pyruvate formate lyase-encoding genes were obtained using this knockout strategy. Double-knockout mutants were also obtained by deleting the citrate lyase-, β-galactosidase-, and aconitase-encoding genes in the pyruvate formate lyase knockout mutant strain. PMID:24610845

Development of a markerless knockout method for Actinobacillus succinogenes.

PubMed

Joshi, Rajasi V; Schindler, Bryan D; McPherson, Nikolas R; Tiwari, Kanupriya; Vieille, Claire

2014-05-01

Actinobacillus succinogenes is one of the best natural succinate-producing organisms, but it still needs engineering to further increase succinate yield and productivity. In this study, we developed a markerless knockout method for A. succinogenes using natural transformation or electroporation. The Escherichia coli isocitrate dehydrogenase gene with flanking flippase recognition target sites was used as the positive selection marker, making use of A. succinogenes's auxotrophy for glutamate to select for growth on isocitrate. The Saccharomyces cerevisiae flippase recombinase (Flp) was used to remove the selection marker, allowing its reuse. Finally, the plasmid expressing flp was cured using acridine orange. We demonstrate that at least two consecutive deletions can be introduced into the same strain using this approach, that no more than a total of 1 kb of DNA is needed on each side of the selection cassette to protect from exonuclease activity during transformation, and that no more than 200 bp of homologous DNA is needed on each side for efficient recombination. We also demonstrate that electroporation can be used as an alternative transformation method to obtain knockout mutants and that an enriched defined medium can be used for direct selection of knockout mutants on agar plates with high efficiency. Single-knockout mutants of the fumarate reductase and of the pyruvate formate lyase-encoding genes were obtained using this knockout strategy. Double-knockout mutants were also obtained by deleting the citrate lyase-, β-galactosidase-, and aconitase-encoding genes in the pyruvate formate lyase knockout mutant strain.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hazelaar, Colien, E-mail: c.hazelaar@vumc.nl; Dahele, Max; Mostafavi, Hassan

Purpose: Spine stereotactic body radiation therapy (SBRT) requires highly accurate positioning. We report our experience with markerless template matching and triangulation of kilovoltage images routinely acquired during spine SBRT, to determine spine position. Methods and Materials: Kilovoltage images, continuously acquired at 7, 11 or 15 frames/s during volumetric modulated spine SBRT of 18 patients, consisting of 93 fluoroscopy datasets (1 dataset/arc), were analyzed off-line. Four patients were immobilized in a head/neck mask, 14 had no immobilization. Two-dimensional (2D) templates were created for each gantry angle from planning computed tomography data and registered to prefiltered kilovoltage images to determine 2D shiftsmore » between actual and planned spine position. Registrations were considered valid if the normalized cross correlation score was ≥0.15. Multiple registrations were triangulated to determine 3D position. For each spine position dataset, average positional offset and standard deviation were calculated. To verify the accuracy and precision of the technique, mean positional offset and standard deviation for twenty stationary phantom datasets with different baseline shifts were measured. Results: For the phantom, average standard deviations were 0.18 mm for left-right (LR), 0.17 mm for superior-inferior (SI), and 0.23 mm for the anterior-posterior (AP) direction. Maximum difference in average detected and applied shift was 0.09 mm. For the 93 clinical datasets, the percentage of valid matched frames was, on average, 90.7% (range: 49.9-96.1%) per dataset. Average standard deviations for all datasets were 0.28, 0.19, and 0.28 mm for LR, SI, and AP, respectively. Spine position offsets were, on average, −0.05 (range: −1.58 to 2.18), −0.04 (range: −3.56 to 0.82), and −0.03 mm (range: −1.16 to 1.51), respectively. Average positional deviation was <1 mm in all directions in 92% of the arcs. Conclusions: Template matching and triangulation using kilovoltage images acquired during irradiation allows spine position detection with submillimeter accuracy at subsecond intervals. Although the majority of patients were not immobilized, most vertebrae were stable at the sub-mm level during spine SBRT delivery.« less

Clinically Normal Stereopsis Does Not Ensure Performance Benefit from Stereoscopic 3D Depth Cues

DTIC Science & Technology

2014-10-28

Stereopsis, Binocular Vision, Optometry , Depth Perception, 3D vision, 3D human factors, Stereoscopic displays, S3D, Virtual environment 16...Binocular Vision, Optometry , Depth Perception, 3D vision, 3D human factors, Stereoscopic displays, S3D, Virtual environment 1 Distribution A: Approved

MEDIASSIST: medical assistance for intraoperative skill transfer in minimally invasive surgery using augmented reality

NASA Astrophysics Data System (ADS)

Sudra, Gunther; Speidel, Stefanie; Fritz, Dominik; Müller-Stich, Beat Peter; Gutt, Carsten; Dillmann, Rüdiger

2007-03-01

Minimally invasive surgery is a highly complex medical discipline with various risks for surgeon and patient, but has also numerous advantages on patient-side. The surgeon has to adapt special operation-techniques and deal with difficulties like the complex hand-eye coordination, limited field of view and restricted mobility. To alleviate with these new problems, we propose to support the surgeon's spatial cognition by using augmented reality (AR) techniques to directly visualize virtual objects in the surgical site. In order to generate an intelligent support, it is necessary to have an intraoperative assistance system that recognizes the surgical skills during the intervention and provides context-aware assistance surgeon using AR techniques. With MEDIASSIST we bundle our research activities in the field of intraoperative intelligent support and visualization. Our experimental setup consists of a stereo endoscope, an optical tracking system and a head-mounted-display for 3D visualization. The framework will be used as platform for the development and evaluation of our research in the field of skill recognition and context-aware assistance generation. This includes methods for surgical skill analysis, skill classification, context interpretation as well as assistive visualization and interaction techniques. In this paper we present the objectives of MEDIASSIST and first results in the fields of skill analysis, visualization and multi-modal interaction. In detail we present a markerless instrument tracking for surgical skill analysis as well as visualization techniques and recognition of interaction gestures in an AR environment.

3D Data Acquisition Platform for Human Activity Understanding

DTIC Science & Technology

2016-03-02

address fundamental research problems of representation and invariant description of3D data, human motion modeling and applications of human activity analysis, and computational optimization of large-scale 3D data.

3D Data Acquisition Platform for Human Activity Understanding

DTIC Science & Technology

2016-03-02

address fundamental research problems of representation and invariant description of 3D data, human motion modeling and applications of human activity analysis, and computational optimization of large-scale 3D data.

Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium

PubMed Central

Zhang, Xinglin; Paganelli, Fernanda L.; Bierschenk, Damien; Kuipers, Annemarie; Bonten, Marc J. M.; Willems, Rob J. L.; van Schaik, Willem

2012-01-01

Enterococcus faecium has become a nosocomial pathogen of major importance, causing infections that are difficult to treat owing to its multi-drug resistance. In particular, resistance to the β-lactam antibiotic ampicillin has become ubiquitous among clinical isolates. Mutations in the low-affinity penicillin binding protein PBP5 have previously been shown to be important for ampicillin resistance in E. faecium, but the existence of additional resistance determinants has been suggested. Here, we constructed a high-density transposon mutant library in E. faecium and developed a transposon mutant tracking approach termed Microarray-based Transposon Mapping (M-TraM), leading to the identification of a compendium of E. faecium genes that contribute to ampicillin resistance. These genes are part of the core genome of E. faecium, indicating a high potential for E. faecium to evolve towards β-lactam resistance. To validate the M-TraM results, we adapted a Cre-lox recombination system to construct targeted, markerless mutants in E. faecium. We confirmed the role of four genes in ampicillin resistance by the generation of targeted mutants and further characterized these mutants regarding their resistance to lysozyme. The results revealed that ddcP, a gene predicted to encode a low-molecular-weight penicillin binding protein with D-alanyl-D-alanine carboxypeptidase activity, was essential for high-level ampicillin resistance. Furthermore, deletion of ddcP sensitized E. faecium to lysozyme and abolished membrane-associated D,D-carboxypeptidase activity. This study has led to the development of a broadly applicable platform for functional genomic-based studies in E. faecium, and it provides a new perspective on the genetic basis of ampicillin resistance in this organism. PMID:22761597

Metabolism of 20-Hydroxyvitamin D3 and 20,23-Dihydroxyvitamin D3 by Rat and Human CYP24A1

PubMed Central

Tieu, Elaine W.; Li, Wei; Chen, Jianjun; Kim, Tae-Kang; Ma, Dejian; Slominski, Andrzej T.; Tuckey, Robert C.

2015-01-01

CYP11A1 hydroxylates vitamin D3 producing 20S-hydroxyvitamin D3 [20(OH)D3] and 20S,23-dihydroxyvitamin D3 [20,23(OH)2D3] as the major and most characterized metabolites. Both display immuno-regulatory and anti-cancer properties while being non-calcemic. A previous study indicated 20(OH)D3 can be metabolized by rat CYP24A1 to products including 20S,24-dihydroxyvitamin D3 [20,24(OH)2D3] and 20S,25-dihydroxyvitamin D3, with both producing greater inhibition of melanoma colony formation than 20(OH)D3. The aim of this study was to characterize the ability of rat and human CYP24A1 to metabolize 20(OH)D3 and 20,23(OH)2D3. Both isoforms metabolized 20(OH)D3 to the same dihydroxyvitamin D species with no secondary metabolites being observed. Hydroxylation at C24 produced both enantiomers of 20,24(OH)2D3. For rat CYP24A1 the preferred initial site of hydroxylation was at C24 whereas the human enzyme preferred C25. 20,23(OH)2D3 was initially metabolized to 20S,23,24-trihydroxyvitamin D3 and 20S,23,25-trihydroxyvitamin D3 by rat and human CYP24A1 as determined by NMR, with both isoforms showing a preference for initial hydroxylation at C25. CYP24A1 was able to further oxidize these metabolites in a series of reactions which included the cleavage of C23-C24 bond, as indicated by high resolution mass spectrometry of the products, analogous to the catabolism of 1,25(OH)2D3 via the C24-oxidation pathway. Similar catalytic efficiencies were observed for the metabolism of 20(OH)D3 and 20,23(OH)2D3 by human CYP24A1 and were lower than for the metabolism of 1,25(OH)2D3. We conclude that rat and human CYP24A1 metabolizes 20(OH)D3 producing only dihydroxyvitamin D3 species as products which retain biological activity, whereas 20,23(OH)2D3 undergoes multiple oxidations which include cleavage of the side chain. PMID:25727742

Using dual-energy x-ray imaging to enhance automated lung tumor tracking during real-time adaptive radiotherapy.

PubMed

Menten, Martin J; Fast, Martin F; Nill, Simeon; Oelfke, Uwe

2015-12-01

Real-time, markerless localization of lung tumors with kV imaging is often inhibited by ribs obscuring the tumor and poor soft-tissue contrast. This study investigates the use of dual-energy imaging, which can generate radiographs with reduced bone visibility, to enhance automated lung tumor tracking for real-time adaptive radiotherapy. kV images of an anthropomorphic breathing chest phantom were experimentally acquired and radiographs of actual lung cancer patients were Monte-Carlo-simulated at three imaging settings: low-energy (70 kVp, 1.5 mAs), high-energy (140 kVp, 2.5 mAs, 1 mm additional tin filtration), and clinical (120 kVp, 0.25 mAs). Regular dual-energy images were calculated by weighted logarithmic subtraction of high- and low-energy images and filter-free dual-energy images were generated from clinical and low-energy radiographs. The weighting factor to calculate the dual-energy images was determined by means of a novel objective score. The usefulness of dual-energy imaging for real-time tracking with an automated template matching algorithm was investigated. Regular dual-energy imaging was able to increase tracking accuracy in left-right images of the anthropomorphic phantom as well as in 7 out of 24 investigated patient cases. Tracking accuracy remained comparable in three cases and decreased in five cases. Filter-free dual-energy imaging was only able to increase accuracy in 2 out of 24 cases. In four cases no change in accuracy was observed and tracking accuracy worsened in nine cases. In 9 out of 24 cases, it was not possible to define a tracking template due to poor soft-tissue contrast regardless of input images. The mean localization errors using clinical, regular dual-energy, and filter-free dual-energy radiographs were 3.85, 3.32, and 5.24 mm, respectively. Tracking success was dependent on tumor position, tumor size, imaging beam angle, and patient size. This study has highlighted the influence of patient anatomy on the success rate of real-time markerless tumor tracking using dual-energy imaging. Additionally, the importance of the spectral separation of the imaging beams used to generate the dual-energy images has been shown.

Using dual-energy x-ray imaging to enhance automated lung tumor tracking during real-time adaptive radiotherapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Menten, Martin J., E-mail: martin.menten@icr.ac.uk; Fast, Martin F.; Nill, Simeon

2015-12-15

Purpose: Real-time, markerless localization of lung tumors with kV imaging is often inhibited by ribs obscuring the tumor and poor soft-tissue contrast. This study investigates the use of dual-energy imaging, which can generate radiographs with reduced bone visibility, to enhance automated lung tumor tracking for real-time adaptive radiotherapy. Methods: kV images of an anthropomorphic breathing chest phantom were experimentally acquired and radiographs of actual lung cancer patients were Monte-Carlo-simulated at three imaging settings: low-energy (70 kVp, 1.5 mAs), high-energy (140 kVp, 2.5 mAs, 1 mm additional tin filtration), and clinical (120 kVp, 0.25 mAs). Regular dual-energy images were calculated bymore » weighted logarithmic subtraction of high- and low-energy images and filter-free dual-energy images were generated from clinical and low-energy radiographs. The weighting factor to calculate the dual-energy images was determined by means of a novel objective score. The usefulness of dual-energy imaging for real-time tracking with an automated template matching algorithm was investigated. Results: Regular dual-energy imaging was able to increase tracking accuracy in left–right images of the anthropomorphic phantom as well as in 7 out of 24 investigated patient cases. Tracking accuracy remained comparable in three cases and decreased in five cases. Filter-free dual-energy imaging was only able to increase accuracy in 2 out of 24 cases. In four cases no change in accuracy was observed and tracking accuracy worsened in nine cases. In 9 out of 24 cases, it was not possible to define a tracking template due to poor soft-tissue contrast regardless of input images. The mean localization errors using clinical, regular dual-energy, and filter-free dual-energy radiographs were 3.85, 3.32, and 5.24 mm, respectively. Tracking success was dependent on tumor position, tumor size, imaging beam angle, and patient size. Conclusions: This study has highlighted the influence of patient anatomy on the success rate of real-time markerless tumor tracking using dual-energy imaging. Additionally, the importance of the spectral separation of the imaging beams used to generate the dual-energy images has been shown.« less

1,25-dihydroxyvitamin D3 induces CCR10 expression in terminally differentiating human B cells.

PubMed

Shirakawa, Aiko-Konno; Nagakubo, Daisuke; Hieshima, Kunio; Nakayama, Takashi; Jin, Zhe; Yoshie, Osamu

2008-03-01

In the B cell lineage, CCR10 is known to be selectively expressed by plasma cells, especially those secreting IgA. In this study, we examined the regulation of CCR10 expression in terminally differentiating human B cells. As reported previously, IL-21 efficiently induced the differentiation of activated human CD19+ B cells into IgD-CD38+ plasma cells in vitro. A minor proportion of the resulting CD19+IgD-CD38+ cells expressed CCR10 at low levels. 1,25-Dihydroxyvitamin D3 (1,25-(OH)2D3), the active metabolite of vitamine D3, dramatically increased the proportion of CD19+IgD-CD38+ cells expressing high levels of CCR10. The 1,25-(OH)2D3 also increased the number of CCR10+ cells expressing surface IgA, although the majority of CCR10+ cells remained negative for surface IgA. Thus, 1,25-(OH)2D3 alone may not be sufficient for the induction of IgA expression in terminally differentiating human B cells. To further determine whether 1,25-(OH)2D3 directly induces CCR10 expression in terminally differentiating B cells, we next performed the analysis on the human CCR10 promoter. We identified a proximal Ets-1 site and an upstream potential vitamin D response element to be critical for the inducible expression of CCR10 by 1,25-(OH)2D3. We confirmed the specific binding of Ets-1 and 1,25-(OH)2D3-activated vitamin D receptor to the respective sites. In conclusion, 1,25-(OH)2D3 efficiently induces CCR10 expression in terminally differentiating human B cells in vitro. Furthermore, the human CCR10 promoter is cooperatively activated by Ets-1 and vitamin D receptor in the presence of 1,25-(OH)2D3.

Comparison of the functional potencies of ropinirole and other dopamine receptor agonists at human D2(long), D3 and D4.4 receptors expressed in Chinese hamster ovary cells

PubMed Central

Coldwell, Martyn C; Boyfield, Izzy; Brown, Tony; Hagan, Jim J; Middlemiss, Derek N

1999-01-01

The aim of the present study was to characterize functional responses to ropinirole, its major metabolites in man (SKF-104557 (4-[2-(propylamino)ethyl]-2-(3H) indolone), SKF-97930 (4-carboxy-2-(3H) indolone)) and other dopamine receptor agonists at human dopamine D2(long) (hD2), D3 (hD3) and D4.4 (hD4) receptors separately expressed in Chinese hamster ovary cells using microphysiometry.All the receptor agonists tested (ropinirole, SKF-104557, SKF-97930, bromocriptine, lisuride, pergolide, pramipexole, talipexole, dopamine) increased extracellular acidification rate in Chinese hamster ovary clones expressing the human D2, D3 or D4 receptor. The pEC50s of ropinirole at hD2, hD3 and hD4 receptors were 7.4, 8.4 and 6.8, respectively. Ropinirole is therefore at least 10 fold selective for the human dopamine D3 receptor over the other D2 receptor family members.At the hD2 and hD3 dopamine receptors all the compounds tested were full agonists as compared to quinpirole. Talipexole and the ropinirole metabolite, SKF-104557, were partial agonists at the hD4 receptor.Bromocriptine and lisuride had a slow onset of agonist action which precluded determination of EC50s.The rank order of agonist potencies was dissimilar to the rank order of radioligand binding affinities at each of the dopamine receptor subtypes. Functional selectivities of the dopamine receptor agonists, as measured in the microphysiometer, were less than radioligand binding selectivities.The results show that ropinirole is a full agonist at human D2, D3 and D4 dopamine receptors. SKF-104557 the major human metabolite of ropinirole, had similar radioligand binding affinities to, but lower functional potencies than, the parent compound. PMID:10455328

Towards Kilo-Hertz 6-DoF Visual Tracking Using an Egocentric Cluster of Rolling Shutter Cameras.

PubMed

Bapat, Akash; Dunn, Enrique; Frahm, Jan-Michael

2016-11-01

To maintain a reliable registration of the virtual world with the real world, augmented reality (AR) applications require highly accurate, low-latency tracking of the device. In this paper, we propose a novel method for performing this fast 6-DOF head pose tracking using a cluster of rolling shutter cameras. The key idea is that a rolling shutter camera works by capturing the rows of an image in rapid succession, essentially acting as a high-frequency 1D image sensor. By integrating multiple rolling shutter cameras on the AR device, our tracker is able to perform 6-DOF markerless tracking in a static indoor environment with minimal latency. Compared to state-of-the-art tracking systems, this tracking approach performs at significantly higher frequency, and it works in generalized environments. To demonstrate the feasibility of our system, we present thorough evaluations on synthetically generated data with tracking frequencies reaching 56.7 kHz. We further validate the method's accuracy on real-world images collected from a prototype of our tracking system against ground truth data using standard commodity GoPro cameras capturing at 120 Hz frame rate.

SU-E-J-59: Feasibility of Markerless Tumor Tracking by Sequential Dual-Energy Fluoroscopy On a Clinical Tumor Tracking System

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dhont, J; Poels, K; Verellen, D

2015-06-15

Purpose: To evaluate the feasibility of markerless tumor tracking through the implementation of a novel dual-energy imaging approach into the clinical dynamic tracking (DT) workflow of the Vero SBRT system. Methods: Two sequential 20 s (11 Hz) fluoroscopy sequences were acquired at the start of one fraction for 7 patients treated for primary and metastatic lung cancer with DT on the Vero system. Sequences were acquired using 2 on-board kV imaging systems located at ±45° from the MV beam axis, at respectively 60 kVp (3.2 mAs) and 120 kVp (2.0 mAs). Offline, a normalized cross-correlation algorithm was applied to matchmore » the high (HE) and low energy (LE) images. Per breathing phase (inhale, exhale, maximum inhale and maximum exhale), the 5 best-matching HE and LE couples were extracted for DE subtraction. A contrast analysis according to gross tumor volume was conducted based on contrast-to-noise ratio (CNR). Improved tumor visibility was quantified using an improvement ratio. Results: Using the implanted fiducial as a benchmark, HE-LE sequence matching was effective for 13 out of 14 imaging angles. Overlying bony anatomy was removed on all DE images. With the exception of two imaging angles, the DE images showed no significantly improved tumor visibility compared to HE images, with an improvement ratio averaged over all patients of 1.46 ± 1.64. Qualitatively, it was observed that for those imaging angles that showed no significantly improved CNR, the tumor tissue could not be reliably visualized on neither HE nor DE images due to a total or partial overlap with other soft tissue. Conclusion: Dual-energy subtraction imaging by sequential orthogonal fluoroscopy was shown feasible by implementing an additional LE fluoroscopy sequence. However, for most imaging angles, DE images did not provide improved tumor visibility over single-energy images. Optimizing imaging angles is likely to improve tumor visibility and the efficacy of dual-energy imaging. This work was in part sponsored by corporate funding from BrainLAB AG.(BrainLAB AG, Feldkirchen, Germany)« less

Markerless attenuation correction for carotid MRI surface receiver coils in combined PET/MR imaging

NASA Astrophysics Data System (ADS)

Eldib, Mootaz; Bini, Jason; Robson, Philip M.; Calcagno, Claudia; Faul, David D.; Tsoumpas, Charalampos; Fayad, Zahi A.

2015-06-01

The purpose of the study was to evaluate the effect of attenuation of MR coils on quantitative carotid PET/MR exams. Additionally, an automated attenuation correction method for flexible carotid MR coils was developed and evaluated. The attenuation of the carotid coil was measured by imaging a uniform water phantom injected with 37 MBq of 18F-FDG in a combined PET/MR scanner for 24 min with and without the coil. In the same session, an ultra-short echo time (UTE) image of the coil on top of the phantom was acquired. Using a combination of rigid and non-rigid registration, a CT-based attenuation map was registered to the UTE image of the coil for attenuation and scatter correction. After phantom validation, the effect of the carotid coil attenuation and the attenuation correction method were evaluated in five subjects. Phantom studies indicated that the overall loss of PET counts due to the coil was 6.3% with local region-of-interest (ROI) errors reaching up to 18.8%. Our registration method to correct for attenuation from the coil decreased the global error and local error (ROI) to 0.8% and 3.8%, respectively. The proposed registration method accurately captured the location and shape of the coil with a maximum spatial error of 2.6 mm. Quantitative analysis in human studies correlated with the phantom findings, but was dependent on the size of the ROI used in the analysis. MR coils result in significant error in PET quantification and thus attenuation correction is needed. The proposed strategy provides an operator-free method for attenuation and scatter correction for a flexible MRI carotid surface coil for routine clinical use.

Cell reprogramming by 3D bioprinting of human fibroblasts in polyurethane hydrogel for fabrication of neural-like constructs.

PubMed

Ho, Lin; Hsu, Shan-Hui

2018-04-01

3D bioprinting is a technique which enables the direct printing of biodegradable materials with cells into 3D tissue. So far there is no cell reprogramming in situ performed with the 3D bioprinting process. Forkhead box D3 (FoxD3) is a transcription factor and neural crest marker, which was reported to reprogram human fibroblasts into neural crest stem-like cells. In this study, we synthesized a new biodegradable thermo-responsive waterborne polyurethane (PU) gel as a bioink. FoxD3 plasmids and human fibroblasts were co-extruded with the PU hydrogel through the syringe needle tip for cell reprogramming. The rheological properties of the PU hydrogel including the modulus, gelation time, and shear thinning were optimized for the transfection effect of FoxD3 in situ. The corresponding shear rate and shear stress were examined. Results showed that human fibroblasts could be reprogrammed into neural crest stem-like cells with high cell viability during the extrusion process under an average shear stress ∼190 Pa. We further translated the method to the extrusion-based 3D bioprinting, and demonstrated that human fibroblasts co-printed with FoxD3 in the thermo-responsive PU hydrogel could be reprogrammed and differentiated into a neural-tissue like construct at 14 days after induction. The neural-like tissue construct produced by 3D bioprinting from human fibroblasts may be applied to personalized drug screening or neuroregeneration. There is no study so far on cell reprogramming in situ with 3D bioprinting. In this manuscript, a new thermoresponsive polyurethane bioink was developed and employed to deliver FoxD3 plasmid into human fibroblasts by the extrusion-based bioprinting. When the polyurethane gel was extruded through the syringe tip, the shear stress generated may have caused the transient membrane permeability for transfection. The shear stress was optimized for transfection in situ by 3D bioprinting. We demonstrated that human fibroblasts could be reprogrammed into neural crest-like stem cells by 3D bioprinting with the gel, and the reprogrammed cells underwent neural differentiation in the printed structure after induction. The neural-like tissue engineering constructs fabricated by 3D bioprinting from human fibroblasts may be applied for neuroregeneration or further developed as mini-brain for basic research and drug screening. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Human perception considerations for 3D content creation

NASA Astrophysics Data System (ADS)

Green, G. Almont

2011-03-01

Observation and interviews with people viewing autostereoscopic 3D imagery provides evidence that there are many human perception considerations required for 3D content creation. A study was undertaken whereby it was witnessed that certain test autostereoscopic imagery elicited a highly emotional response and engagement, while other test autostereoscopic imagery was given only a passing glance. That an image can be viewed with a certain level of stereopsis does not make it compelling. By taking into consideration the manner in which humans perceive depth and the space between objects, 3D content can achieve a level of familiarity and realness that is not possible with single perspective imagery. When human perception issues are ignored, 3D imagery can be undesirable to viewers and a negative bias against 3D imagery can occur. The preparation of 3D content is more important than the display technology. Where human perception, as it is used to interpret reality, is not mimicked in the creation of 3D content, the general public typically express a negative bias against that imagery (where choices are provided). For some, the viewing of 3D content that could not exist naturally, induces physical discomfort.

Sexual Dimorphism Analysis and Gender Classification in 3D Human Face

NASA Astrophysics Data System (ADS)

Hu, Yuan; Lu, Li; Yan, Jingqi; Liu, Zhi; Shi, Pengfei

In this paper, we present the sexual dimorphism analysis in 3D human face and perform gender classification based on the result of sexual dimorphism analysis. Four types of features are extracted from a 3D human-face image. By using statistical methods, the existence of sexual dimorphism is demonstrated in 3D human face based on these features. The contributions of each feature to sexual dimorphism are quantified according to a novel criterion. The best gender classification rate is 94% by using SVMs and Matcher Weighting fusion method.This research adds to the knowledge of 3D faces in sexual dimorphism and affords a foundation that could be used to distinguish between male and female in 3D faces.

Quantitative targeted absolute proteomic analysis of transporters, receptors and junction proteins for validation of human cerebral microvascular endothelial cell line hCMEC/D3 as a human blood-brain barrier model.

PubMed

Ohtsuki, Sumio; Ikeda, Chiemi; Uchida, Yasuo; Sakamoto, Yumi; Miller, Florence; Glacial, Fabienne; Decleves, Xavier; Scherrmann, Jean-Michel; Couraud, Pierre-Olivier; Kubo, Yoshiyuki; Tachikawa, Masanori; Terasaki, Tetsuya

2013-01-07

Human cerebral microvascular endothelial cell line hCMEC/D3 is an established model of the human blood-brain barrier (BBB). The purpose of the present study was to determine, by means of quantitative targeted absolute proteomics, the protein expression levels in hCMEC/D3 cells of multiple transporters, receptors and junction proteins for comparison with our previously reported findings in isolated human brain microvessels. Among 91 target molecules, 12 transporters, 2 receptors, 1 junction protein and 1 membrane marker were present at quantifiable levels in plasma membrane fraction of hCMEC/D3 cells. ABCA2, MDR1, MRP4, BCRP, GLUT1, 4F2hc, MCT1, ENT1, transferrin and insulin receptors and claudin-5 were detected in both hCMEC/D3 cells and human brain microvessels. After normalization based on Na(+)/K(+) ATPase expression, the differences in protein expression levels between hCMEC/D3 cells and human brain microvessels were within 4-fold for these proteins, with the exceptions of ENT1, transferrin receptor and claudin-5. ABCA8, LAT1, LRP1 and γ-GTP were below the limit of quantification in the cells, but were found in human brain microvessels. ABCA3, ABCA6, MRP1 and ATA1 were found only in hCMEC/D3 cells. Furthermore, compared with human umbilical vein endothelial cells (HUVECs) as reference nonbrain endothelial cells, MDR1 was found only in hCMEC/D3 cells, and GLUT1 expression was 15-fold higher in hCMEC/D3 cells than in HUVECs. In conclusion, this is the first study to examine the suitability and limitations of the hCMEC/D3 cell line as a BBB functional model in terms of quantitative expression levels of transporters, receptors and tight junction proteins.

Learning dictionaries of sparse codes of 3D movements of body joints for real-time human activity understanding.

PubMed

Qi, Jin; Yang, Zhiyong

2014-01-01

Real-time human activity recognition is essential for human-robot interactions for assisted healthy independent living. Most previous work in this area is performed on traditional two-dimensional (2D) videos and both global and local methods have been used. Since 2D videos are sensitive to changes of lighting condition, view angle, and scale, researchers begun to explore applications of 3D information in human activity understanding in recently years. Unfortunately, features that work well on 2D videos usually don't perform well on 3D videos and there is no consensus on what 3D features should be used. Here we propose a model of human activity recognition based on 3D movements of body joints. Our method has three steps, learning dictionaries of sparse codes of 3D movements of joints, sparse coding, and classification. In the first step, space-time volumes of 3D movements of body joints are obtained via dense sampling and independent component analysis is then performed to construct a dictionary of sparse codes for each activity. In the second step, the space-time volumes are projected to the dictionaries and a set of sparse histograms of the projection coefficients are constructed as feature representations of the activities. Finally, the sparse histograms are used as inputs to a support vector machine to recognize human activities. We tested this model on three databases of human activities and found that it outperforms the state-of-the-art algorithms. Thus, this model can be used for real-time human activity recognition in many applications.

Effects of 1,25-dihydroxyvitamin D3 and vitamin D3 on the expression of the vitamin D receptor in human skeletal muscle cells

USDA-ARS?s Scientific Manuscript database

Vitamin D receptor (VDR) expression and action in non-human skeletal muscle have recently been reported in several studies, yet data on the activity and expression of VDR in human muscle cells are scarce. We conducted a series of studies to examine the (1) effect of 1,25-dihydroxyvitamin D3 (1,25(OH...

Effect of viewing distance on 3D fatigue caused by viewing mobile 3D content

NASA Astrophysics Data System (ADS)

Mun, Sungchul; Lee, Dong-Su; Park, Min-Chul; Yano, Sumio

2013-05-01

With an advent of autostereoscopic display technique and increased needs for smart phones, there has been a significant growth in mobile TV markets. The rapid growth in technical, economical, and social aspects has encouraged 3D TV manufacturers to apply 3D rendering technology to mobile devices so that people have more opportunities to come into contact with many 3D content anytime and anywhere. Even if the mobile 3D technology leads to the current market growth, there is an important thing to consider for consistent development and growth in the display market. To put it briefly, human factors linked to mobile 3D viewing should be taken into consideration before developing mobile 3D technology. Many studies have investigated whether mobile 3D viewing causes undesirable biomedical effects such as motion sickness and visual fatigue, but few have examined main factors adversely affecting human health. Viewing distance is considered one of the main factors to establish optimized viewing environments from a viewer's point of view. Thus, in an effort to determine human-friendly viewing environments, this study aims to investigate the effect of viewing distance on human visual system when exposing to mobile 3D environments. Recording and analyzing brainwaves before and after watching mobile 3D content, we explore how viewing distance affects viewing experience from physiological and psychological perspectives. Results obtained in this study are expected to provide viewing guidelines for viewers, help ensure viewers against undesirable 3D effects, and lead to make gradual progress towards a human-friendly mobile 3D viewing.

Uniform Local Binary Pattern Based Texture-Edge Feature for 3D Human Behavior Recognition.

PubMed

Ming, Yue; Wang, Guangchao; Fan, Chunxiao

2015-01-01

With the rapid development of 3D somatosensory technology, human behavior recognition has become an important research field. Human behavior feature analysis has evolved from traditional 2D features to 3D features. In order to improve the performance of human activity recognition, a human behavior recognition method is proposed, which is based on a hybrid texture-edge local pattern coding feature extraction and integration of RGB and depth videos information. The paper mainly focuses on background subtraction on RGB and depth video sequences of behaviors, extracting and integrating historical images of the behavior outlines, feature extraction and classification. The new method of 3D human behavior recognition has achieved the rapid and efficient recognition of behavior videos. A large number of experiments show that the proposed method has faster speed and higher recognition rate. The recognition method has good robustness for different environmental colors, lightings and other factors. Meanwhile, the feature of mixed texture-edge uniform local binary pattern can be used in most 3D behavior recognition.

Graphic and movie illustrations of human prenatal development and their application to embryological education based on the human embryo specimens in the Kyoto collection.

PubMed

Yamada, Shigehito; Uwabe, Chigako; Nakatsu-Komatsu, Tomoko; Minekura, Yutaka; Iwakura, Masaji; Motoki, Tamaki; Nishimiya, Kazuhiko; Iiyama, Masaaki; Kakusho, Koh; Minoh, Michihiko; Mizuta, Shinobu; Matsuda, Tetsuya; Matsuda, Yoshimasa; Haishi, Tomoyuki; Kose, Katsumi; Fujii, Shingo; Shiota, Kohei

2006-02-01

Morphogenesis in the developing embryo takes place in three dimensions, and in addition, the dimension of time is another important factor in development. Therefore, the presentation of sequential morphological changes occurring in the embryo (4D visualization) is essential for understanding the complex morphogenetic events and the underlying mechanisms. Until recently, 3D visualization of embryonic structures was possible only by reconstruction from serial histological sections, which was tedious and time-consuming. During the past two decades, 3D imaging techniques have made significant advances thanks to the progress in imaging and computer technologies, computer graphics, and other related techniques. Such novel tools have enabled precise visualization of the 3D topology of embryonic structures and to demonstrate spatiotemporal 4D sequences of organogenesis. Here, we describe a project in which staged human embryos are imaged by the magnetic resonance (MR) microscope, and 3D images of embryos and their organs at each developmental stage were reconstructed based on the MR data, with the aid of computer graphics techniques. On the basis of the 3D models of staged human embryos, we constructed a data set of 3D images of human embryos and made movies to illustrate the sequential process of human morphogenesis. Furthermore, a computer-based self-learning program of human embryology is being developed for educational purposes, using the photographs, histological sections, MR images, and 3D models of staged human embryos. Copyright 2005 Wiley-Liss, Inc.

Markerless gating for lung cancer radiotherapy based on machine learning techniques

NASA Astrophysics Data System (ADS)

Lin, Tong; Li, Ruijiang; Tang, Xiaoli; Dy, Jennifer G.; Jiang, Steve B.

2009-03-01

In lung cancer radiotherapy, radiation to a mobile target can be delivered by respiratory gating, for which we need to know whether the target is inside or outside a predefined gating window at any time point during the treatment. This can be achieved by tracking one or more fiducial markers implanted inside or near the target, either fluoroscopically or electromagnetically. However, the clinical implementation of marker tracking is limited for lung cancer radiotherapy mainly due to the risk of pneumothorax. Therefore, gating without implanted fiducial markers is a promising clinical direction. We have developed several template-matching methods for fluoroscopic marker-less gating. Recently, we have modeled the gating problem as a binary pattern classification problem, in which principal component analysis (PCA) and support vector machine (SVM) are combined to perform the classification task. Following the same framework, we investigated different combinations of dimensionality reduction techniques (PCA and four nonlinear manifold learning methods) and two machine learning classification methods (artificial neural networks—ANN and SVM). Performance was evaluated on ten fluoroscopic image sequences of nine lung cancer patients. We found that among all combinations of dimensionality reduction techniques and classification methods, PCA combined with either ANN or SVM achieved a better performance than the other nonlinear manifold learning methods. ANN when combined with PCA achieves a better performance than SVM in terms of classification accuracy and recall rate, although the target coverage is similar for the two classification methods. Furthermore, the running time for both ANN and SVM with PCA is within tolerance for real-time applications. Overall, ANN combined with PCA is a better candidate than other combinations we investigated in this work for real-time gated radiotherapy.

Evaluation of a portable markerless finger position capture device: accuracy of the Leap Motion controller in healthy adults.

PubMed

Tung, James Y; Lulic, Tea; Gonzalez, Dave A; Tran, Johnathan; Dickerson, Clark R; Roy, Eric A

2015-05-01

Although motion analysis is frequently employed in upper limb motor assessment (e.g. visually-guided reaching), they are resource-intensive and limited to laboratory settings. This study evaluated the reliability and accuracy of a new markerless motion capture device, the Leap Motion controller, to measure finger position. Testing conditions that influence reliability and agreement between the Leap and a research-grade motion capture system were examined. Nine healthy young adults pointed to 15 targets on a computer screen under two conditions: (1) touching the target (touch) and (2) 4 cm away from the target (no-touch). Leap data was compared to an Optotrak marker attached to the index finger. Across all trials, root mean square (RMS) error of the Leap system was 17.30  ±  9.56 mm (mean ± SD), sampled at 65.47  ±  21.53 Hz. The % viable trials and mean sampling rate were significantly lower in the touch condition (44% versus 64%, p < 0.001; 52.02  ±  2.93 versus 73.98  ±  4.48 Hz, p = 0.003). While linear correlations were high (horizontal: r(2) = 0.995, vertical r(2) = 0.945), the limits of agreement were large (horizontal: -22.02 to +26.80 mm, vertical: -29.41 to +30.14 mm). While not as precise as more sophisticated optical motion capture systems, the Leap Motion controller is sufficiently reliable for measuring motor performance in pointing tasks that do not require high positional accuracy (e.g. reaction time, Fitt's, trails, bimanual coordination).

Modeling human diseases with induced pluripotent stem cells: from 2D to 3D and beyond.

PubMed

Liu, Chun; Oikonomopoulos, Angelos; Sayed, Nazish; Wu, Joseph C

2018-03-08

The advent of human induced pluripotent stem cells (iPSCs) presents unprecedented opportunities to model human diseases. Differentiated cells derived from iPSCs in two-dimensional (2D) monolayers have proven to be a relatively simple tool for exploring disease pathogenesis and underlying mechanisms. In this Spotlight article, we discuss the progress and limitations of the current 2D iPSC disease-modeling platform, as well as recent advancements in the development of human iPSC models that mimic in vivo tissues and organs at the three-dimensional (3D) level. Recent bioengineering approaches have begun to combine different 3D organoid types into a single '4D multi-organ system'. We summarize the advantages of this approach and speculate on the future role of 4D multi-organ systems in human disease modeling. © 2018. Published by The Company of Biologists Ltd.

3D culture models of Alzheimer's disease: a road map to a "cure-in-a-dish".

PubMed

Choi, Se Hoon; Kim, Young Hye; Quinti, Luisa; Tanzi, Rudolph E; Kim, Doo Yeon

2016-12-09

Alzheimer's disease (AD) transgenic mice have been used as a standard AD model for basic mechanistic studies and drug discovery. These mouse models showed symbolic AD pathologies including β-amyloid (Aβ) plaques, gliosis and memory deficits but failed to fully recapitulate AD pathogenic cascades including robust phospho tau (p-tau) accumulation, clear neurofibrillary tangles (NFTs) and neurodegeneration, solely driven by familial AD (FAD) mutation(s). Recent advances in human stem cell and three-dimensional (3D) culture technologies made it possible to generate novel 3D neural cell culture models that recapitulate AD pathologies including robust Aβ deposition and Aβ-driven NFT-like tau pathology. These new 3D human cell culture models of AD hold a promise for a novel platform that can be used for mechanism studies in human brain-like environment and high-throughput drug screening (HTS). In this review, we will summarize the current progress in recapitulating AD pathogenic cascades in human neural cell culture models using AD patient-derived induced pluripotent stem cells (iPSCs) or genetically modified human stem cell lines. We will also explain how new 3D culture technologies were applied to accelerate Aβ and p-tau pathologies in human neural cell cultures, as compared the standard two-dimensional (2D) culture conditions. Finally, we will discuss a potential impact of the human 3D human neural cell culture models on the AD drug-development process. These revolutionary 3D culture models of AD will contribute to accelerate the discovery of novel AD drugs.

Regulation of 1-alpha, 25-dihydroxyvitamin D3 on interleukin-6 and interleukin-8 induced by sulfur mustard (HD) on human skin cells.

PubMed

Arroyo, Carmen M; Kan, Robert K; Burman, Damon L; Kahler, David W; Nelson, Marian R; Corun, Charlene M; Guzman, Juanita J; Broomfield, Clarence A

2003-05-01

The regulatory effects of the active form of vitamin D, 1-alpha, 25-dihydroxyvitamin D3 (1-alpha, 25 (OH)2D3) were assessed on the cytokine and chemokine secretion induced by sulfur mustard on human skin fibroblasts and human epidermal keratinocytes. Stimulation of human skin fibroblasts with sulfur mustard (10(-4) M for 24 hr at 37 degrees ) resulted in approximately a 5 times increase in the secretion of interleukin-6 and over a 10 times increase for interleukin-8, which was inhibited by 1-alpha, 25 (OH)2D3, at

Proof-of-concept: 3D bioprinting of pigmented human skin constructs.

PubMed

Ng, Wei Long; Qi, Jovina Tan Zhi; Yeong, Wai Yee; Naing, May Win

2018-01-23

Three-dimensional (3D) pigmented human skin constructs have been fabricated using a 3D bioprinting approach. The 3D pigmented human skin constructs are obtained from using three different types of skin cells (keratinocytes, melanocytes and fibroblasts from three different skin donors) and they exhibit similar constitutive pigmentation (pale pigmentation) as the skin donors. A two-step drop-on-demand bioprinting strategy facilitates the deposition of cell droplets to emulate the epidermal melanin units (pre-defined patterning of keratinocytes and melanocytes at the desired positions) and manipulation of the microenvironment to fabricate 3D biomimetic hierarchical porous structures found in native skin tissue. The 3D bioprinted pigmented skin constructs are compared to the pigmented skin constructs fabricated by conventional a manual-casting approach; in-depth characterization of both the 3D pigmented skin constructs has indicated that the 3D bioprinted skin constructs have a higher degree of resemblance to native skin tissue in term of the presence of well-developed stratified epidermal layers and the presence of a continuous layer of basement membrane proteins as compared to the manually-cast samples. The 3D bioprinting approach facilitates the development of 3D in vitro pigmented human skin constructs for potential toxicology testing and fundamental cell biology research.

Studying the Brain in a Dish: 3D Cell Culture Models of Human Brain Development and Disease.

PubMed

Brown, Juliana; Quadrato, Giorgia; Arlotta, Paola

2018-01-01

The study of the cellular and molecular processes of the developing human brain has been hindered by access to suitable models of living human brain tissue. Recently developed 3D cell culture models offer the promise of studying fundamental brain processes in the context of human genetic background and species-specific developmental mechanisms. Here, we review the current state of 3D human brain organoid models and consider their potential to enable investigation of complex aspects of human brain development and the underpinning of human neurological disease. © 2018 Elsevier Inc. All rights reserved.

Estimating Physical Activity Energy Expenditure with the Kinect Sensor in an Exergaming Environment

PubMed Central

Nathan, David; Huynh, Du Q.; Rubenson, Jonas; Rosenberg, Michael

2015-01-01

Active video games that require physical exertion during game play have been shown to confer health benefits. Typically, energy expended during game play is measured using devices attached to players, such as accelerometers, or portable gas analyzers. Since 2010, active video gaming technology incorporates marker-less motion capture devices to simulate human movement into game play. Using the Kinect Sensor and Microsoft SDK this research aimed to estimate the mechanical work performed by the human body and estimate subsequent metabolic energy using predictive algorithmic models. Nineteen University students participated in a repeated measures experiment performing four fundamental movements (arm swings, standing jumps, body-weight squats, and jumping jacks). Metabolic energy was captured using a Cortex Metamax 3B automated gas analysis system with mechanical movement captured by the combined motion data from two Kinect cameras. Estimations of the body segment properties, such as segment mass, length, centre of mass position, and radius of gyration, were calculated from the Zatsiorsky-Seluyanov's equations of de Leva, with adjustment made for posture cost. GPML toolbox implementation of the Gaussian Process Regression, a locally weighted k-Nearest Neighbour Regression, and a linear regression technique were evaluated for their performance on predicting the metabolic cost from new feature vectors. The experimental results show that Gaussian Process Regression outperformed the other two techniques by a small margin. This study demonstrated that physical activity energy expenditure during exercise, using the Kinect camera as a motion capture system, can be estimated from segmental mechanical work. Estimates for high-energy activities, such as standing jumps and jumping jacks, can be made accurately, but for low-energy activities, such as squatting, the posture of static poses should be considered as a contributing factor. When translated into the active video gaming environment, the results could be incorporated into game play to more accurately control the energy expenditure requirements. PMID:26000460

Tranylcypromine Substituted cis-Hydroxycyclobutylnaphthamides as Potent and Selective Dopamine D3 Receptor Antagonists

PubMed Central

2015-01-01

We report a class of potent and selective dopamine D3 receptor antagonists based upon tranylcypromine. Although tranylcypromine has a low affinity for the rat D3 receptor (Ki = 12.8 μM), our efforts have yielded (1R,2S)-11 (CJ-1882), which has Ki values of 2.7 and 2.8 nM at the rat and human dopamine D3 receptors, respectively, and displays respective selectivities of >10000-fold and 223-fold over the rat and human D2 receptors. Evaluation in a β-arrestin functional assay showed that (1R,2S)-11 is a potent and competitive antagonist at the human D3 receptor. PMID:24848155

Vitamin D3 analog maxacalcitol (OCT) induces hCAP-18/LL-37 production in human oral epithelial cells.

PubMed

Tada, Hiroyuki; Shimizu, Takamitsu; Nagaoka, Isao; Takada, Haruhiko

2016-01-01

Maxacalcitol (22-oxacalcitriol: OCT) is a synthetic vitamin D3 analog with a limited calcemic effect. In this study, we investigated whether OCT increases the production of LL-37/CAP-18, a human cathelicidin antimicrobial peptide, in human gingival/oral epithelial cells. A human gingival epithelial cell line (Ca9-22) and human oral epithelial cell lines (HSC-2, HSC-3, and HSC-4) exhibited the enhanced expression of LL-37 mRNA upon stimulation with OCT as well as active metabolites of vitamins D3 and D2. Among the human epithelial cell lines, Ca9-22 exhibited the strongest response to these vitamin D-related compounds. OCT induced the higher production of CAP-18 (ng/mL order) until 6 days time-dependently in Ca9-22 cells in culture. The periodontal pathogen Porphyromonas gingivalis was killed by treatment with the LL-37 peptide. These findings suggest that OCT induces the production of hCAP-18/LL-37 in a manner similar to that induced by the active metabolite of vitamin D3.

3D Human Motion Editing and Synthesis: A Survey

PubMed Central

Wang, Xin; Chen, Qiudi; Wang, Wanliang

2014-01-01

The ways to compute the kinematics and dynamic quantities of human bodies in motion have been studied in many biomedical papers. This paper presents a comprehensive survey of 3D human motion editing and synthesis techniques. Firstly, four types of methods for 3D human motion synthesis are introduced and compared. Secondly, motion capture data representation, motion editing, and motion synthesis are reviewed successively. Finally, future research directions are suggested. PMID:25045395

Real-time 3D human pose recognition from reconstructed volume via voxel classifiers

NASA Astrophysics Data System (ADS)

Yoo, ByungIn; Choi, Changkyu; Han, Jae-Joon; Lee, Changkyo; Kim, Wonjun; Suh, Sungjoo; Park, Dusik; Kim, Junmo

2014-03-01

This paper presents a human pose recognition method which simultaneously reconstructs a human volume based on ensemble of voxel classifiers from a single depth image in real-time. The human pose recognition is a difficult task since a single depth camera can capture only visible surfaces of a human body. In order to recognize invisible (self-occluded) surfaces of a human body, the proposed algorithm employs voxel classifiers trained with multi-layered synthetic voxels. Specifically, ray-casting onto a volumetric human model generates a synthetic voxel, where voxel consists of a 3D position and ID corresponding to the body part. The synthesized volumetric data which contain both visible and invisible body voxels are utilized to train the voxel classifiers. As a result, the voxel classifiers not only identify the visible voxels but also reconstruct the 3D positions and the IDs of the invisible voxels. The experimental results show improved performance on estimating the human poses due to the capability of inferring the invisible human body voxels. It is expected that the proposed algorithm can be applied to many fields such as telepresence, gaming, virtual fitting, wellness business, and real 3D contents control on real 3D displays.

Understanding Human Perception of Building Categories in Virtual 3d Cities - a User Study

NASA Astrophysics Data System (ADS)

Tutzauer, P.; Becker, S.; Niese, T.; Deussen, O.; Fritsch, D.

2016-06-01

Virtual 3D cities are becoming increasingly important as a means of visually communicating diverse urban-related information. To get a deeper understanding of a human's cognitive experience of virtual 3D cities, this paper presents a user study on the human ability to perceive building categories (e.g. residential home, office building, building with shops etc.) from geometric 3D building representations. The study reveals various dependencies between geometric properties of the 3D representations and the perceptibility of the building categories. Knowledge about which geometries are relevant, helpful or obstructive for perceiving a specific building category is derived. The importance and usability of such knowledge is demonstrated based on a perception-guided 3D building abstraction process.

Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats.

PubMed

Richert, Lysiane; Baze, Audrey; Parmentier, Céline; Gerets, Helga H J; Sison-Young, Rowena; Dorau, Martina; Lovatt, Cerys; Czich, Andreas; Goldring, Christopher; Park, B Kevin; Juhila, Satu; Foster, Alison J; Williams, Dominic P

2016-09-06

Sixteen training compounds selected in the IMI MIP-DILI consortium, 12 drug-induced liver injury (DILI) positive compounds and 4 non-DILI compounds, were assessed in cryopreserved primary human hepatocytes. When a ten-fold safety margin threshold was applied, the non-DILI-compounds were correctly identified 2h following a single exposure to pooled human hepatocytes (n=13 donors) in suspension and 14-days following repeat dose exposure (3 treatments) to an established 3D-microtissue co-culture (3D-MT co-culture, n=1 donor) consisting of human hepatocytes co-cultured with non-parenchymal cells (NPC). In contrast, only 5/12 DILI-compounds were correctly identified 2h following a single exposure to pooled human hepatocytes in suspension. Exposure of the 2D-sandwich culture human hepatocyte monocultures (2D-sw) for 3days resulted in the correct identification of 11/12 DILI-positive compounds, whereas exposure of the human 3D-MT co-cultures for 14days resulted in identification of 9/12 DILI-compounds; in addition to ximelagatran (also not identified by 2D-sw monocultures, Sison-Young et al., 2016), the 3D-MT co-cultures failed to detect amiodarone and bosentan. The sensitivity of the 2D human hepatocytes co-cultured with NPC to ximelagatran was increased in the presence of lipopolysaccharide (LPS), but only at high concentrations, therefore preventing its classification as a DILI positive compound. In conclusion (1) despite suspension human hepatocytes having the greatest metabolic capacity in the short term, they are the least predictive of clinical DILI across the MIP-DILI test compounds, (2) longer exposure periods than 72h of human hepatocytes do not allow to increase DILI-prediction rate, (3) co-cultures of human hepatocytes with NPC, in the presence of LPS during the 72h exposure period allow the assessment of innate immune system involvement of a given drug. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

An Inducible Cytochrome P450 3A4-Dependent Vitamin D Catabolic Pathway

PubMed Central

Wang, Zhican; Lin, Yvonne S.; Zheng, Xi Emily; Senn, Tauri; Hashizume, Takanori; Scian, Michele; Dickmann, Leslie J.; Nelson, Sidney D.; Baillie, Thomas A.; Hebert, Mary F.; Blough, David; Davis, Connie L.

2012-01-01

Vitamin D3 is critical for the regulation of calcium and phosphate homeostasis. In some individuals, mineral homeostasis can be disrupted by long-term therapy with certain antiepileptic drugs and the antimicrobial agent rifampin, resulting in drug-induced osteomalacia, which is attributed to vitamin D deficiency. We now report a novel CYP3A4-dependent pathway, the 4-hydroxylation of 25-hydroxyvitamin D3 (25OHD3), the induction of which may contribute to drug-induced vitamin D deficiency. The metabolism of 25OHD3 was fully characterized in vitro. CYP3A4 was the predominant source of 25OHD3 hydroxylation by human liver microsomes, with the formation of 4β,25-dihydroxyvitamin D3 [4β,25(OH)2D3] dominating (Vmax/Km = 0.85 ml · min−1 · nmol enzyme−1). 4β,25(OH)2D3 was found in human plasma at concentrations comparable to that of 1α,25-dihydroxyvitamin D3, and its formation rate in a panel of human liver microsomes was strongly correlated with CYP3A4 content and midazolam hydroxylation activity. Formation of 4β,25(OH)2D3 in primary human hepatocytes was induced by rifampin and inhibited by CYP3A4-specific inhibitors. Short-term treatment of healthy volunteers (n = 6) with rifampin selectively induced CYP3A4-dependent 4β,25(OH)2D3, but not CYP24A1-dependent 24R,25-dihydroxyvitamin D3 formation, and altered systemic mineral homeostasis. Our results suggest that CYP3A4-dependent 25OHD3 metabolism may play an important role in the regulation of vitamin D3 in vivo and in the etiology of drug-induced osteomalacia. PMID:22205755

A Measure of the Effectiveness of Incorporating 3D Human Anatomy into an Online Undergraduate Laboratory

ERIC Educational Resources Information Center

Hilbelink, Amy J.

2009-01-01

Results of a study designed to determine the effectiveness of implementing three-dimensional (3D) stereo images of a human skull in an undergraduate human anatomy online laboratory were gathered and analysed. Mental model theory and its applications to 3D relationships are discussed along with the research results. Quantitative results on 62 pairs…

Image-based tracking of the suturing needle during laparoscopic interventions

NASA Astrophysics Data System (ADS)

Speidel, S.; Kroehnert, A.; Bodenstedt, S.; Kenngott, H.; Müller-Stich, B.; Dillmann, R.

2015-03-01

One of the most complex and difficult tasks for surgeons during minimally invasive interventions is suturing. A prerequisite to assist the suturing process is the tracking of the needle. The endoscopic images provide a rich source of information which can be used for needle tracking. In this paper, we present an image-based method for markerless needle tracking. The method uses a color-based and geometry-based segmentation to detect the needle. Once an initial needle detection is obtained, a region of interest enclosing the extracted needle contour is passed on to a reduced segmentation. It is evaluated with in vivo images from da Vinci interventions.

3D Reconstruction of Static Human Body with a Digital Camera

NASA Astrophysics Data System (ADS)

Remondino, Fabio

2003-01-01

Nowadays the interest in 3D reconstruction and modeling of real humans is one of the most challenging problems and a topic of great interest. The human models are used for movies, video games or ergonomics applications and they are usually created with 3D scanner devices. In this paper a new method to reconstruct the shape of a static human is presented. Our approach is based on photogrammetric techniques and uses a sequence of images acquired around a standing person with a digital still video camera or with a camcorder. First the images are calibrated and orientated using a bundle adjustment. After the establishment of a stable adjusted image block, an image matching process is performed between consecutive triplets of images. Finally the 3D coordinates of the matched points are computed with a mean accuracy of ca 2 mm by forward ray intersection. The obtained point cloud can then be triangulated to generate a surface model of the body or a virtual human model can be fitted to the recovered 3D data. Results of the 3D human point cloud with pixel color information are presented.

2D and 3D Traveling Salesman Problem

ERIC Educational Resources Information Center

Haxhimusa, Yll; Carpenter, Edward; Catrambone, Joseph; Foldes, David; Stefanov, Emil; Arns, Laura; Pizlo, Zygmunt

2011-01-01

When a two-dimensional (2D) traveling salesman problem (TSP) is presented on a computer screen, human subjects can produce near-optimal tours in linear time. In this study we tested human performance on a real and virtual floor, as well as in a three-dimensional (3D) virtual space. Human performance on the real floor is as good as that on a…

Three-dimensional contractile muscle tissue consisting of human skeletal myocyte cell line.

PubMed

Shima, Ai; Morimoto, Yuya; Sweeney, H Lee; Takeuchi, Shoji

2018-06-18

This paper describes a method to construct three-dimensional (3D) contractile human skeletal muscle tissues from a cell line. The 3D tissue was fabricated as a fiber-based structure and cultured for two weeks under tension by anchoring its both ends. While myotubes from the immortalized human skeletal myocytes used in this study never contracted in the conventional two-dimensional (2D) monolayer culture, myotubes in the 3D tissue showed spontaneous contraction at a high frequency and also reacted to the electrical stimulation. Immunofluorescence revealed that the myotubes in the 3D tissues had sarcomeres and expressed ryanodine receptor (RyR) and sarco/endoplasmic reticulum Ca 2+ -ATPase (SERCA). In addition, intracellular calcium oscillations in the myotubes in the 3D tissue were observed. These results indicated that the 3D culture enabled the myocyte cell line to reach a more highly matured state compared to 2D culture. Since contraction is the most significant feature of skeletal muscle, we believe that our 3D human muscle tissue with the contractile ability would be a useful tool for both basic biology research and drug discovery as one of the muscle-on-chips. Copyright © 2018. Published by Elsevier Inc.

Volumetric 3D display using a DLP projection engine

NASA Astrophysics Data System (ADS)

Geng, Jason

2012-03-01

In this article, we describe a volumetric 3D display system based on the high speed DLPTM (Digital Light Processing) projection engine. Existing two-dimensional (2D) flat screen displays often lead to ambiguity and confusion in high-dimensional data/graphics presentation due to lack of true depth cues. Even with the help of powerful 3D rendering software, three-dimensional (3D) objects displayed on a 2D flat screen may still fail to provide spatial relationship or depth information correctly and effectively. Essentially, 2D displays have to rely upon capability of human brain to piece together a 3D representation from 2D images. Despite the impressive mental capability of human visual system, its visual perception is not reliable if certain depth cues are missing. In contrast, volumetric 3D display technologies to be discussed in this article are capable of displaying 3D volumetric images in true 3D space. Each "voxel" on a 3D image (analogous to a pixel in 2D image) locates physically at the spatial position where it is supposed to be, and emits light from that position toward omni-directions to form a real 3D image in 3D space. Such a volumetric 3D display provides both physiological depth cues and psychological depth cues to human visual system to truthfully perceive 3D objects. It yields a realistic spatial representation of 3D objects and simplifies our understanding to the complexity of 3D objects and spatial relationship among them.

3D imaging of cleared human skin biopsies using light-sheet microscopy: A new way to visualize in-depth skin structure.

PubMed

Abadie, S; Jardet, C; Colombelli, J; Chaput, B; David, A; Grolleau, J-L; Bedos, P; Lobjois, V; Descargues, P; Rouquette, J

2018-05-01

Human skin is composed of the superimposition of tissue layers of various thicknesses and components. Histological staining of skin sections is the benchmark approach to analyse the organization and integrity of human skin biopsies; however, this approach does not allow 3D tissue visualization. Alternatively, confocal or two-photon microscopy is an effective approach to perform fluorescent-based 3D imaging. However, owing to light scattering, these methods display limited light penetration in depth. The objectives of this study were therefore to combine optical clearing and light-sheet fluorescence microscopy (LSFM) to perform in-depth optical sectioning of 5 mm-thick human skin biopsies and generate 3D images of entire human skin biopsies. A benzyl alcohol and benzyl benzoate solution was used to successfully optically clear entire formalin fixed human skin biopsies, making them transparent. In-depth optical sectioning was performed with LSFM on the basis of tissue-autofluorescence observations. 3D image analysis of optical sections generated with LSFM was performed by using the Amira ® software. This new approach allowed us to observe in situ the different layers and compartments of human skin, such as the stratum corneum, the dermis and epidermal appendages. With this approach, we easily performed 3D reconstruction to visualise an entire human skin biopsy. Finally, we demonstrated that this method is useful to visualise and quantify histological anomalies, such as epidermal hyperplasia. The combination of optical clearing and LSFM has new applications in dermatology and dermatological research by allowing 3D visualization and analysis of whole human skin biopsies. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Conformational Changes in the Carpus During Finger Traps Distraction

PubMed Central

Leventhal, Evan L.; Moore, Douglas C.; Akelman, Edward; Wolfe, Scott W.; Crisco, Joseph J.

2010-01-01

Introduction Wrist distraction is a common treatment maneuver used clinically for the reduction of distal radial fractures and mid-carpal dislocations. Wrist distraction is also required during wrist arthroscopy to access the radiocarpal joint and has been used as a test for scapholunate ligament injury. However, the effect of a distraction load on the normal wrist has not been well studied. The purpose of this study was to measure the 3-D conformational changes of the carpal bones in the normal wrist as a result of a static distractive load. Methods The dominant wrists of 14 healthy volunteers were scanned using computed tomography at rest and during application of 98N of distraction. Load was applied using finger traps and volunteers were encouraged to relax their forearm muscles and to allow distraction of the wrist. The motions of the bones in the wrist were tracked between the unloaded and loaded trial using markerless bone registration. The average displacement vector of each bone was calculated relative to the radius as well as the interbone distances for 20 bone-bone interactions. Joint separation was estimated at the radiocarpal, midcarpal and carpal-metacarpal joints in the direction of loading using the radius, lunate, capitate and 3rd metacarpal. Results With loading, the distance between the radius and 3rd metacarpal increased an average of 3.3±3.1mm in the direction of loading. This separation was primarily located in the axial direction at the radiocarpal (1.0±1.0mm) and midcarpal (2.0±1.7mm) joints. There were minimal changes in the transverse direction within the distal row, although the proximal row narrowed by 0.98±0.7mm. Distraction between the radius and scaphoid (2.5±2.2mm) was 2.4 times greater than between the radius and lunate (1.0±1.0mm). Conclusions Carpal distraction has a significant effect on the conformation of the carpus, especially at the radiocarpal and midcarpal joints. In the normal wrist, external traction causes twice as much distraction at the lunocapitate joint than at the radiolunate joint. PMID:20141894

Evaluation of 3D-human skin equivalents for assessment of human dermal absorption of some brominated flame retardants.

PubMed

Abdallah, Mohamed Abou-Elwafa; Pawar, Gopal; Harrad, Stuart

2015-11-01

Ethical and technical difficulties inherent to studies in human tissues are impeding assessment of the dermal bioavailability of brominated flame retardants (BFRs). This is further complicated by increasing restrictions on the use of animals in toxicity testing, and the uncertainties associated with extrapolating data from animal studies to humans due to inter-species variations. To overcome these difficulties, we evaluate 3D-human skin equivalents (3D-HSE) as a novel in vitro alternative to human and animal testing for assessment of dermal absorption of BFRs. The percutaneous penetration of hexabromocyclododecanes (HBCD) and tetrabromobisphenol-A (TBBP-A) through two commercially available 3D-HSE models was studied and compared to data obtained for human ex vivo skin according to a standard protocol. No statistically significant differences were observed between the results obtained using 3D-HSE and human ex vivo skin at two exposure levels. The absorbed dose was low (less than 7%) and was significantly correlated with log Kow of the tested BFR. Permeability coefficient values showed increasing dermal resistance to the penetration of γ-HBCD>β-HBCD>α-HBCD>TBBPA. The estimated long lag times (>30 min) suggests that frequent hand washing may reduce human exposure to HBCDs and TBBPA via dermal contact. Copyright © 2015 Elsevier Ltd. All rights reserved.

3D Data Acquisition Platform for Human Activity Understanding

DTIC Science & Technology

2016-03-02

3D data. The support for the acquisition of such research instrumentation have significantly facilitated our current and future research and educate ...SECURITY CLASSIFICATION OF: In this project, we incorporated motion capture devices, 3D vision sensors, and EMG sensors to cross validate...multimodality data acquisition, and address fundamental research problems of representation and invariant description of 3D data, human motion modeling and

3D Miniaturization of Human Organs for Drug Discovery.

PubMed

Park, Joseph; Wetzel, Isaac; Dréau, Didier; Cho, Hansang

2018-01-01

"Engineered human organs" hold promises for predicting the effectiveness and accuracy of drug responses while reducing cost, time, and failure rates in clinical trials. Multiorgan human models utilize many aspects of currently available technologies including self-organized spherical 3D human organoids, microfabricated 3D human organ chips, and 3D bioprinted human organ constructs to mimic key structural and functional properties of human organs. They enable precise control of multicellular activities, extracellular matrix (ECM) compositions, spatial distributions of cells, architectural organizations of ECM, and environmental cues. Thus, engineered human organs can provide the microstructures and biological functions of target organs and advantageously substitute multiscaled drug-testing platforms including the current in vitro molecular assays, cell platforms, and in vivo models. This review provides an overview of advanced innovative designs based on the three main technologies used for organ construction leading to single and multiorgan systems useable for drug development. Current technological challenges and future perspectives are also discussed. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Gait disorder rehabilitation using vision and non-vision based sensors: A systematic review

PubMed Central

Ali, Asraf; Sundaraj, Kenneth; Ahmad, Badlishah; Ahamed, Nizam; Islam, Anamul

2012-01-01

Even though the amount of rehabilitation guidelines has never been greater, uncertainty continues to arise regarding the efficiency and effectiveness of the rehabilitation of gait disorders. This question has been hindered by the lack of information on accurate measurements of gait disorders. Thus, this article reviews the rehabilitation systems for gait disorder using vision and non-vision sensor technologies, as well as the combination of these. All papers published in the English language between 1990 and June, 2012 that had the phrases “gait disorder” “rehabilitation”, “vision sensor”, or “non vision sensor” in the title, abstract, or keywords were identified from the SpringerLink, ELSEVIER, PubMed, and IEEE databases. Some synonyms of these phrases and the logical words “and” “or” and “not” were also used in the article searching procedure. Out of the 91 published articles found, this review identified 84 articles that described the rehabilitation of gait disorders using different types of sensor technologies. This literature set presented strong evidence for the development of rehabilitation systems using a markerless vision-based sensor technology. We therefore believe that the information contained in this review paper will assist the progress of the development of rehabilitation systems for human gait disorders. PMID:22938548

Brain Human Monoclonal Autoantibody from Sydenham Chorea Targets Dopaminergic Neurons in Transgenic Mice and Signals Dopamine D2 Receptor: Implications in Human Disease1

PubMed Central

Cox, Carol J.; Sharma, Meenakshi; Leckman, James F.; Zuccolo, Jonathan; Zuccolo, Amir; Kovoor, Abraham; Swedo, Susan E.; Cunningham, Madeleine W.

2013-01-01

How autoantibodies target the brain and lead to disease in disorders such as Sydenham chorea (SC) is not known. SC is characterized by autoantibodies against the brain and is the main neurologic manifestation of streptococcal-induced rheumatic fever. Previously, our novel SC-derived mAb 24.3.1 was found to recognize streptococcal and brain antigens. To investigate in vivo targets of human mAb 24.3.1, VH/VL genes were expressed in B cells of transgenic (Tg) mice as functional chimeric human VH 24.3.1 - mouse constant region IgG1a autoantibody. Chimeric human-mouse IgG1a autoantibody co-localized with tyrosine hydroxylase in the basal ganglia within dopaminergic neurons in vivo in VH 24.3.1 Tg mice. Both human mAb 24.3.1 and IgG1a in Tg sera were found to react with human dopamine D2 receptor (D2R). Reactivity of chorea-derived mAb 24.3.1 or SC IgG with D2R was confirmed by 1) dose dependent inhibitory signaling of D2R as a potential consequence of targeting dopaminergic neurons, 2) reaction with surface-exposed FLAG epitope-tagged D2R, and 3) blocking of Ab reactivity by an extracellular D2R peptide. IgG from SC and a related subset of streptococcal associated behavioral disorders called pediatric autoimmune neuropsychiatric disorder associated with streptococci (PANDAS) with small choreiform movements reacted in ELISA with D2R. Reaction with FLAG-tagged D2R distinguished SC from PANDAS while sera from both SC and PANDAS induced inhibitory signaling of D2R on transfected cells comparable to dopamine. Here we define a mechanism by which the brain may be altered by antibody in movement and behavioral disorders. PMID:24184556

MiR-520d-3p antitumor activity in human breast cancer via post-transcriptional regulation of spindle and kinetochore associated 2 expression.

PubMed

Ren, Zhouhui; Yang, Tong; Ding, Jie; Liu, Weihong; Meng, Xiangyu; Zhang, Pingping; Liu, Kaitai; Wang, Ping

2018-01-01

MicroRNAs (miRNAs) play an important role in human tumorigenesis as oncogenes or tumor suppressors by directly binding to the 3'-untranslated region of their target mRNAs. MiR-520d-3p has been reported as a tumor suppressor gene in ovarian cancer and gastric cancer, while the function of miR-520d-3p in human breast cancers is still uninvolved. In this study, we initially identified that the expression of miR-520d-3p was significantly reduced in breast cancer specimens and cell lines. The restoration of miR-520d-3p expression not only reduced breast cancer cell viability by causing the accumulation of G2 phase and cell apoptosis, but also inhibited tumorigenicity in vivo . In addition, as a critical target of miR-520d-3p, the activity of spindle and kinetochore associated 2 (SKA2) was greatly inhibited by miR-520d-3p, and overexpression of miR-520d-3p decreased the expression of SKA2. SKA2 downregulation suppressed cell viability, whereas restoration of SKA2 expression significantly reversed the inhibitory effects of miR-520d-3p antitumor activity. Furthermore, SKA2 was frequently overexpressed in clinical specimens and cell lines, and the expression levels were statistically inversely correlated with miR-520d-3p expression. In conclusion, our data demonstrated that miR-520d-3p antitumor activity is achieved by targeting the SKA2 in human breast cancer cells, suggesting that miR-520d-3p may be a potential target molecule for the therapy.

Foveated model observers to predict human performance in 3D images

NASA Astrophysics Data System (ADS)

Lago, Miguel A.; Abbey, Craig K.; Eckstein, Miguel P.

2017-03-01

We evaluate 3D search requires model observers that take into account the peripheral human visual processing (foveated models) to predict human observer performance. We show that two different 3D tasks, free search and location-known detection, influence the relative human visual detectability of two signals of different sizes in synthetic backgrounds mimicking the noise found in 3D digital breast tomosynthesis. One of the signals resembled a microcalcification (a small and bright sphere), while the other one was designed to look like a mass (a larger Gaussian blob). We evaluated current standard models observers (Hotelling; Channelized Hotelling; non-prewhitening matched filter with eye filter, NPWE; and non-prewhitening matched filter model, NPW) and showed that they incorrectly predict the relative detectability of the two signals in 3D search. We propose a new model observer (3D Foveated Channelized Hotelling Observer) that incorporates the properties of the visual system over a large visual field (fovea and periphery). We show that the foveated model observer can accurately predict the rank order of detectability of the signals in 3D images for each task. Together, these results motivate the use of a new generation of foveated model observers for predicting image quality for search tasks in 3D imaging modalities such as digital breast tomosynthesis or computed tomography.

1,25D3 enhances antitumor activity of gemcitabine and cisplatin in human bladder cancer models

PubMed Central

Ma, Yingyu; Yu, Wei-Dong; Trump, Donald L.; Johnson, Candace S.

2010-01-01

Background 1,25 dihydroxyvitamin D3 (1,25D3) potentiates the cytotoxic effects of several common chemotherapeutic agents. The combination of gemcitabine and cisplatin (GC) is a current standard chemotherapy regimen for bladder cancer. We investigated whether 1,25D3 could enhance the antitumor activity of GC in bladder cancer model systems. Methods Human bladder cancer T24 and UMUC3 cells were pretreated with 1,25D3 followed by GC. Apoptosis were assessed by annexin V staining. Caspase activation was examined by immunoblot analysis and substrate-based caspase activity assay. The cytotoxic effects were examined using MTT and in vitro clonogenic assay. p73 protein levels were assessed by immunoblot analysis. Knockdown of p73 was achieved by siRNA. The in vivo antitumor activity was assessed by in vivo excision clonogenic assay and tumor regrowth delay in the T24 xenograft model. Results 1,25D3 pretreatment enhanced GC-induced apoptosis and the activities of caspases- 8, 9 and 3 in T24 and UMUC3 cells. 1,25D3 synergistically reduced GC-suppressed surviving fraction in T24 cells. 1,25D3, gemcitabine, or cisplatin induced p73 accumulation, which was enhanced by GC or 1,25D3 and GC. p73 expression was lower in human primary bladder tumor tissue compared with adjacent normal tissue. Knockdown of p73 increased clonogenic capacity of T24 cells treated with 1,25D3, GC or 1,25D3 and GC. 1,25D3 and GC combination enhanced tumor regression compared with 1,25D3 or GC alone. Conclusions 1,25D3 potentiates GC-mediated growth inhibition in human bladder cancer models in vitro and in vivo, which involves p73 induction and apoptosis. PMID:20564622

Fabrication and evaluation of electrohydrodynamic jet 3D printed polycaprolactone/chitosan cell carriers using human embryonic stem cell-derived fibroblasts.

PubMed

Wu, Yang; Sriram, Gopu; Fawzy, Amr S; Fuh, Jerry Yh; Rosa, Vinicius; Cao, Tong; Wong, Yoke San

2016-08-01

Biological function of adherent cells depends on the cell-cell and cell-matrix interactions in three-dimensional space. To understand the behavior of cells in 3D environment and their interactions with neighboring cells and matrix requires 3D culture systems. Here, we present a novel 3D cell carrier scaffold that provides an environment for routine 3D cell growth in vitro We have developed thin, mechanically stable electrohydrodynamic jet (E-jet) 3D printed polycaprolactone and polycaprolactone/Chitosan macroporous scaffolds with precise fiber orientation for basic 3D cell culture application. We have evaluated the application of this technology by growing human embryonic stem cell-derived fibroblasts within these 3D scaffolds. Assessment of cell viability and proliferation of cells seeded on polycaprolactone and polycaprolactone/Chitosan 3D-scaffolds show that the human embryonic stem cell-derived fibroblasts could adhere and proliferate on the scaffolds over time. Further, using confocal microscopy we demonstrate the ability to use fluorescence-labelled cells that could be microscopically monitored in real-time. Hence, these 3D printed polycaprolactone and polycaprolactone/Chitosan scaffolds could be used as a cell carrier for in vitro 3D cell culture-, bioreactor- and tissue engineering-related applications in the future. © The Author(s) 2016.

Investigation Of Integrating Three-Dimensional (3-D) Geometry Into The Visual Anatomical Injury Descriptor (Visual AID) Using WebGL

DTIC Science & Technology

2011-08-01

generated using the Zygote Human Anatomy 3-D model (3). Use of a reference anatomy independent of personal identification, such as Zygote, allows Visual...Zygote Human Anatomy 3D Model, 2010. http://www.zygote.com/ (accessed July 26, 2011). 4. Khronos Group Web site. Khronos to Create New Open Standard for...understanding of the information at hand. In order to fulfill the medical illustration track, I completed a concentration in science, focusing on human

Stereo chromatic contrast sensitivity model to blue-yellow gratings.

PubMed

Yang, Jiachen; Lin, Yancong; Liu, Yun

2016-03-07

As a fundamental metric of human visual system (HVS), contrast sensitivity function (CSF) is typically measured by sinusoidal gratings at the detection of thresholds for psychophysically defined cardinal channels: luminance, red-green, and blue-yellow. Chromatic CSF, which is a quick and valid index to measure human visual performance and various retinal diseases in two-dimensional (2D) space, can not be directly applied into the measurement of human stereo visual performance. And no existing perception model considers the influence of chromatic CSF of inclined planes on depth perception in three-dimensional (3D) space. The main aim of this research is to extend traditional chromatic contrast sensitivity characteristics to 3D space and build a model applicable in 3D space, for example, strengthening stereo quality of 3D images. This research also attempts to build a vision model or method to check human visual characteristics of stereo blindness. In this paper, CRT screen was clockwise and anti-clockwise rotated respectively to form the inclined planes. Four inclined planes were selected to investigate human chromatic vision in 3D space and contrast threshold of each inclined plane was measured with 18 observers. Stimuli were isoluminant blue-yellow sinusoidal gratings. Horizontal spatial frequencies ranged from 0.05 to 5 c/d. Contrast sensitivity was calculated as the inverse function of the pooled cone contrast threshold. According to the relationship between spatial frequency of inclined plane and horizontal spatial frequency, the chromatic contrast sensitivity characteristics in 3D space have been modeled based on the experimental data. The results show that the proposed model can well predicted human chromatic contrast sensitivity characteristics in 3D space.

Generation of a chickenized catalytic anti-nucleic acid antibody by complementarity-determining region grafting.

PubMed

Roh, Jooho; Byun, Sung June; Seo, Youngsil; KIm, Minjae; Lee, Jae-Ho; Kim, Songmi; Lee, Yuno; Lee, Keun Woo; Kim, Jin-Kyoo; Kwon, Myung-Hee

2015-02-01

In contrast to a number of studies on the humanization of non-human antibodies, the reshaping of a non-human antibody into a chicken antibody has never been attempted. Therefore, nothing is known about the animal species-dependent compatibility of the framework regions (FRs) that sustain the appropriate conformation of the complementarity-determining regions (CDRs). In this study, we attempted the reshaping of the variable domains of the mouse catalytic anti-nucleic acid antibody 3D8 (m3D8) into the FRs of a chicken antibody (“chickenization”) by CDR grafting, which is a common method for the humanization of antibodies. CDRs of the acceptor chicken antibody that showed a high homology to the FRs of m3D8 were replaced with those of m3D8, resulting in the chickenized antibody (ck3D8). ck3D8 retained the biochemical properties (DNA binding, DNA hydrolysis, and cellular internalizing activities) and three-dimensional structure of m3D8 and showed reduced immunogenicity in chickens. Our study demonstrates that CDR grafting can be applied to the chickenization of a mouse antibody, probably due to the interspecies compatibility of the FRs.

Understanding 3D human torso shape via manifold clustering

NASA Astrophysics Data System (ADS)

Li, Sheng; Li, Peng; Fu, Yun

2013-05-01

Discovering the variations in human torso shape plays a key role in many design-oriented applications, such as suit designing. With recent advances in 3D surface imaging technologies, people can obtain 3D human torso data that provide more information than traditional measurements. However, how to find different human shapes from 3D torso data is still an open problem. In this paper, we propose to use spectral clustering approach on torso manifold to address this problem. We first represent high-dimensional torso data in a low-dimensional space using manifold learning algorithm. Then the spectral clustering method is performed to get several disjoint clusters. Experimental results show that the clusters discovered by our approach can describe the discrepancies in both genders and human shapes, and our approach achieves better performance than the compared clustering method.

Comparison of Image Generation And Processing Techniques For 3D Reconstruction of The Human Skull

DTIC Science & Technology

2001-10-25

inexpensive Microscribe (3D digitizer) with a standard widely used and expensive CT-Scan and/or MRI for 3D reconstruction of a human skull, which will be... Microscribe 3D digitizing unit and another one using the CT-Scans (2D cross-sections) obtained from a GE scanner. Both models were then subjected to stress...these methods are still elaborate, expensive and not readily accessible. Using the hand-held digitizer, the Microscribe , X, Y and Z coordinates

Midbrain-like Organoids from Human Pluripotent Stem Cells Contain Functional Dopaminergic and Neuromelanin-Producing Neurons.

PubMed

Jo, Junghyun; Xiao, Yixin; Sun, Alfred Xuyang; Cukuroglu, Engin; Tran, Hoang-Dai; Göke, Jonathan; Tan, Zi Ying; Saw, Tzuen Yih; Tan, Cheng-Peow; Lokman, Hidayat; Lee, Younghwan; Kim, Donghoon; Ko, Han Seok; Kim, Seong-Oh; Park, Jae Hyeon; Cho, Nam-Joon; Hyde, Thomas M; Kleinman, Joel E; Shin, Joo Heon; Weinberger, Daniel R; Tan, Eng King; Je, Hyunsoo Shawn; Ng, Huck-Hui

2016-08-04

Recent advances in 3D culture systems have led to the generation of brain organoids that resemble different human brain regions; however, a 3D organoid model of the midbrain containing functional midbrain dopaminergic (mDA) neurons has not been reported. We developed a method to differentiate human pluripotent stem cells into a large multicellular organoid-like structure that contains distinct layers of neuronal cells expressing characteristic markers of human midbrain. Importantly, we detected electrically active and functionally mature mDA neurons and dopamine production in our 3D midbrain-like organoids (MLOs). In contrast to human mDA neurons generated using 2D methods or MLOs generated from mouse embryonic stem cells, our human MLOs produced neuromelanin-like granules that were structurally similar to those isolated from human substantia nigra tissues. Thus our MLOs bearing features of the human midbrain may provide a tractable in vitro system to study the human midbrain and its related diseases. Copyright © 2016 Elsevier Inc. All rights reserved.

Using the Microsoft Kinect™ to assess 3-D shoulder kinematics during computer use.

PubMed

Xu, Xu; Robertson, Michelle; Chen, Karen B; Lin, Jia-Hua; McGorry, Raymond W

2017-11-01

Shoulder joint kinematics has been used as a representative indicator to investigate musculoskeletal symptoms among computer users for office ergonomics studies. The traditional measurement of shoulder kinematics normally requires a laboratory-based motion tracking system which limits the field studies. In the current study, a portable, low cost, and marker-less Microsoft Kinect™ sensor was examined for its feasibility on shoulder kinematics measurement during computer tasks. Eleven healthy participants performed a standardized computer task, and their shoulder kinematics data were measured by a Kinect sensor and a motion tracking system concurrently. The results indicated that placing the Kinect sensor in front of the participants would yielded a more accurate shoulder kinematics measurements then placing the Kinect sensor 15° or 30° to one side. The results also showed that the Kinect sensor had a better estimate on shoulder flexion/extension, compared with shoulder adduction/abduction and shoulder axial rotation. The RMSE of front-placed Kinect sensor on shoulder flexion/extension was less than 10° for both the right and the left shoulder. The measurement error of the front-placed Kinect sensor on the shoulder adduction/abduction was approximately 10° to 15°, and the magnitude of error is proportional to the magnitude of that joint angle. After the calibration, the RMSE on shoulder adduction/abduction were less than 10° based on an independent dataset of 5 additional participants. For shoulder axial rotation, the RMSE of front-placed Kinect sensor ranged between approximately 15° to 30°. The results of the study suggest that the Kinect sensor can provide some insight on shoulder kinematics for improving office ergonomics. Copyright © 2017 Elsevier Ltd. All rights reserved.

Human factors guidelines for applications of 3D perspectives: a literature review

NASA Astrophysics Data System (ADS)

Dixon, Sharon; Fitzhugh, Elisabeth; Aleva, Denise

2009-05-01

Once considered too processing-intense for general utility, application of the third dimension to convey complex information is facilitated by the recent proliferation of technological advancements in computer processing, 3D displays, and 3D perspective (2.5D) renderings within a 2D medium. The profusion of complex and rapidly-changing dynamic information being conveyed in operational environments has elevated interest in possible military applications of 3D technologies. 3D can be a powerful mechanism for clearer information portrayal, facilitating rapid and accurate identification of key elements essential to mission performance and operator safety. However, implementation of 3D within legacy systems can be costly, making integration prohibitive. Therefore, identifying which tasks may benefit from 3D or 2.5D versus simple 2D visualizations is critical. Unfortunately, there is no "bible" of human factors guidelines for usability optimization of 2D, 2.5D, or 3D visualizations nor for determining which display best serves a particular application. Establishing such guidelines would provide an invaluable tool for designers and operators. Defining issues common to each will enhance design effectiveness. This paper presents the results of an extensive review of open source literature addressing 3D information displays, with particular emphasis on comparison of true 3D with 2D and 2.5D representations and their utility for military tasks. Seventy-five papers are summarized, highlighting militarily relevant applications of 3D visualizations and 2.5D perspective renderings. Based on these findings, human factors guidelines for when and how to use these visualizations, along with recommendations for further research are discussed.

Femur Model Reconstruction Based on Reverse Engineering and Rapid Prototyping

NASA Astrophysics Data System (ADS)

Tang, Tongming; Zhang, Zheng; Ni, Hongjun; Deng, Jiawen; Huang, Mingyu

Precise reconstruction of 3D models is fundamental and crucial to the researches of human femur. In this paper we present our approach towards tackling this problem. The surface of a human femur was scanned using a hand-held 3D laser scanner. The data obtained, in the form of point cloud, was then processed using the reverse engineering software Geomagic and the CAD/CAM software CimatronE to reconstruct a digital 3D model. The digital model was then used by the rapid prototyping machine to build a physical model of human femur using 3D printing. The geometric characteristics of the obtained physical model matched that of the original femur. The process of "physical object - 3D data - digital 3D model - physical model" presented in this paper provides a foundation of precise modeling for the digital manufacturing, virtual assembly, stress analysis, and simulated surgery of artificial bionic femurs.

Growth of human breast tissues from patient cells in 3D hydrogel scaffolds.

PubMed

Sokol, Ethan S; Miller, Daniel H; Breggia, Anne; Spencer, Kevin C; Arendt, Lisa M; Gupta, Piyush B

2016-03-01

Three-dimensional (3D) cultures have proven invaluable for expanding human tissues for basic research and clinical applications. In both contexts, 3D cultures are most useful when they (1) support the outgrowth of tissues from primary human cells that have not been immortalized through extensive culture or viral infection and (2) include defined, physiologically relevant components. Here we describe a 3D culture system with both of these properties that stimulates the outgrowth of morphologically complex and hormone-responsive mammary tissues from primary human breast epithelial cells. Primary human breast epithelial cells isolated from patient reduction mammoplasty tissues were seeded into 3D hydrogels. The hydrogel scaffolds were composed of extracellular proteins and carbohydrates present in human breast tissue and were cultured in serum-free medium containing only defined components. The physical properties of these hydrogels were determined using atomic force microscopy. Tissue growth was monitored over time using bright-field and fluorescence microscopy, and maturation was assessed using morphological metrics and by immunostaining for markers of stem cells and differentiated cell types. The hydrogel tissues were also studied by fabricating physical models from confocal images using a 3D printer. When seeded into these 3D hydrogels, primary human breast epithelial cells rapidly self-organized in the absence of stromal cells and within 2 weeks expanded to form mature mammary tissues. The mature tissues contained luminal, basal, and stem cells in the correct topological orientation and also exhibited the complex ductal and lobular morphologies observed in the human breast. The expanded tissues became hollow when treated with estrogen and progesterone, and with the further addition of prolactin produced lipid droplets, indicating that they were responding to hormones. Ductal branching was initiated by clusters of cells expressing putative mammary stem cell markers, which subsequently localized to the leading edges of the tissue outgrowths. Ductal elongation was preceded by leader cells that protruded from the tips of ducts and engaged with the extracellular matrix. These 3D hydrogel scaffolds support the growth of complex mammary tissues from primary patient-derived cells. We anticipate that this culture system will empower future studies of human mammary gland development and biology.

High throughput LC-MS/MS method for the simultaneous analysis of multiple vitamin D analytes in serum.

PubMed

Jenkinson, Carl; Taylor, Angela E; Hassan-Smith, Zaki K; Adams, John S; Stewart, Paul M; Hewison, Martin; Keevil, Brian G

2016-03-01

Recent studies suggest that vitamin D-deficiency is linked to increased risk of common human health problems. To define vitamin D 'status' most routine analytical methods quantify one particular vitamin D metabolite, 25-hydroxyvitamin D3 (25OHD3). However, vitamin D is characterized by complex metabolic pathways, and simultaneous measurement of multiple vitamin D metabolites may provide a more accurate interpretation of vitamin D status. To address this we developed a high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to analyse multiple vitamin D analytes, with particular emphasis on the separation of epimer metabolites. A supportive liquid-liquid extraction (SLE) and LC-MS/MS method was developed to quantify 10 vitamin D metabolites as well as separation of an interfering 7α-hydroxy-4-cholesten-3-one (7αC4) isobar (precursor of bile acid), and validated by analysis of human serum samples. In a cohort of 116 healthy subjects, circulating concentrations of 25-hydroxyvitamin D3 (25OHD3), 3-epi-25-hydroxyvitamin D3 (3-epi-25OHD3), 24,25-dihydroxyvitamin D3 (24R,25(OH)2D3), 1,25-dihydroxyvitamin D3 (1α,25(OH)2D3), and 25-hydroxyvitamin D2 (25OHD2) were quantifiable using 220μL of serum, with 25OHD3 and 24R,25(OH)2D3 showing significant seasonal variations. This high-throughput LC-MS/MS method provides a novel strategy for assessing the impact of vitamin D on human health and disease. Copyright © 2016 Elsevier B.V. All rights reserved.

Study of optical design of three-dimensional digital ophthalmoscopes.

PubMed

Fang, Yi-Chin; Yen, Chih-Ta; Chu, Chin-Hsien

2015-10-01

This study primarily involves using optical zoom structures to design a three-dimensional (3D) human-eye optical sensory system with infrared and visible light. According to experimental data on two-dimensional (2D) and 3D images, human-eye recognition of 3D images is substantially higher (approximately 13.182%) than that of 2D images. Thus, 3D images are more effective than 2D images when they are used at work or in high-recognition devices. In the optical system design, infrared and visible light wavebands were incorporated as light sources to perform simulations. The results can be used to facilitate the design of optical systems suitable for 3D digital ophthalmoscopes.

3D tissue-like assemblies: A novel approach to investigate virus-cell interactions.

PubMed

Goodwin, Thomas J; McCarthy, Maureen; Cohrs, Randall J; Kaufer, Benedikt B

2015-11-15

Virus-host cell interactions are most commonly analyzed in cells maintained in vitro as two-dimensional tissue cultures. However, these in vitro conditions vary quite drastically from the tissues that are commonly infected in vivo. Over the years, a number of systems have been developed that allow the establishment of three-dimensional (3D) tissue structures that have properties similar to their in vivo 3D counterparts. These 3D systems have numerous applications including drug testing, maintenance of large tissue explants, monitoring migration of human lymphocytes in tissues, analysis of human organ tissue development and investigation of virus-host interactions including viral latency. Here, we describe the establishment of tissue-like assemblies for human lung and neuronal tissue that we infected with a variety of viruses including the respiratory pathogens human parainfluenza virus type 3 (PIV3), respiratory syncytial virus (RSV) and SARS corona virus (SARS-CoV) as well as the human neurotropic herpesvirus, varicella-zoster virus (VZV). Copyright © 2015 Elsevier Inc. All rights reserved.

3D Tissue-Like Assemblies: A Novel Approach to Investigate Virus-Cell Interactions

PubMed Central

Goodwin, Thomas J.; McCarthy, Maureen; Cohrs, Randall J.; Kaufer, Benedikt B.

2017-01-01

Virus-host cell interactions are most commonly analyzed in cells maintained in vitro as two-dimensional tissue cultures. However, these in vitro conditions vary quite drastically from the tissues that are commonly infected in vivo. Over the years, a number of systems have been developed that allow the establishment of three-dimensional (3D) tissue structures that have properties similar to their in vivo 3D counterparts. These 3D systems have numerous applications including drug testing, maintenance of large tissue explants, monitoring migration of human lymphocytes in tissues, analysis of human organ tissue development and investigation of virus-host interactions including viral latency. Here, we describe the establishment of tissue-like assemblies for human lung and neuronal tissue that we infected with a variety of viruses including the respiratory pathogens human parainfluenza virus type 3 (PIV3), respiratory syncytial virus (RSV) and SARS corona virus (SARS-CoV) as well as the human neurotropic herpesvirus, varicella-zoster virus (VZV) PMID:25986169

Human body motion tracking based on quantum-inspired immune cloning algorithm

NASA Astrophysics Data System (ADS)

Han, Hong; Yue, Lichuan; Jiao, Licheng; Wu, Xing

2009-10-01

In a static monocular camera system, to gain a perfect 3D human body posture is a great challenge for Computer Vision technology now. This paper presented human postures recognition from video sequences using the Quantum-Inspired Immune Cloning Algorithm (QICA). The algorithm included three parts. Firstly, prior knowledge of human beings was used, the key joint points of human could be detected automatically from the human contours and skeletons which could be thinning from the contours; And due to the complexity of human movement, a forecasting mechanism of occlusion joint points was addressed to get optimum 2D key joint points of human body; And then pose estimation recovered by optimizing between the 2D projection of 3D human key joint points and 2D detection key joint points using QICA, which recovered the movement of human body perfectly, because this algorithm could acquire not only the global optimal solution, but the local optimal solution.

D-type cyclins in adult human testis and testicular cancer: relation to cell type, proliferation, differentiation, and malignancy.

PubMed

Bartkova, J; Rajpert-de Meyts, E; Skakkebaek, N E; Bartek, J

1999-04-01

D-type cyclins are proto-oncogenic components of the 'RB pathway', a G1/S regulatory mechanism centred around the retinoblastoma tumour suppressor (pRB) implicated in key cellular decisions that control cell proliferation, cell-cycle arrest, quiescence, and differentiation. This study focused on immunohistochemical and immunochemical analysis of human adult testis and 32 testicular tumours to examine the differential expression and abundance of cyclins D1, D2, and D3 in relation to cell type, proliferation, differentiation, and malignancy. In normal testis, the cell type-restricted expression patterns were dominated by high levels of cyclin D3 in quiescent Leydig cells and the lack of any D-type cyclin in the germ cells, the latter possibly representing the only example of normal mammalian cells proliferating in the absence of these cyclins. Most carcinoma-in-situ lesions appeared to gain expression of cyclin D2 but not D1 or D3, while the invasive testicular tumours showed variable positivity for cyclins D2 and D3, but rarely D1. An unexpected correlation with differentiation rather than proliferation was found particularly for cyclin D3 in teratomas, a conceptually significant observation confirmed by massive up-regulation of cyclin D3 in the human teratocarcinoma cell line NTera2/D1 induced to differentiate along the neuronal lineage. These results suggest a possible involvement of cyclin D2 in the early stages of testicular oncogenesis and the striking examples of proliferation-independent expression point to potential dual or multiple roles of the D-type cyclins, particularly of cyclin D3. These findings extend current concepts of the biology of the cyclin D subfamily, as well as of the biology and oncopathology of the human adult testis. Apart from practical implications for the assessment of proliferation and oncogenic aberrations in human tissues and tumours, this study may inspire further research into the emerging role of the cyclin D proteins in the establishment and/or maintenance of the differentiated phenotypes. Copyright 1999 John Wiley & Sons, Ltd.

Blonanserin extensively occupies rat dopamine D3 receptors at antipsychotic dose range.

PubMed

Baba, Satoko; Enomoto, Takeshi; Horisawa, Tomoko; Hashimoto, Takashi; Ono, Michiko

2015-03-01

Antagonism of the dopamine D3 receptor has been hypothesized to be beneficial for schizophrenia cognitive deficits, negative symptoms and extrapyramidal symptoms. However, recent animal and human studies have shown that most antipsychotics do not occupy D3 receptors in vivo, despite their considerable binding affinity for this receptor in vitro. In the present study, we investigated the D3 receptor binding of blonanserin, a dopamine D2/D3 and serotonin 5-HT2A receptors antagonist, in vitro and in vivo. Blonanserin showed the most potent binding affinity for human D3 receptors among the tested atypical antipsychotics (risperidone, olanzapine and aripiprazole). Our GTPγS-binding assay demonstrated that blonanserin acts as a potent full antagonist for human D3 receptors. All test-drugs exhibited antipsychotic-like efficacy in methamphetamine-induced hyperactivity in rats. Treatment with blonanserin at its effective dose blocked the binding of [(3)H]-(+)-PHNO, a D2/D3 receptor radiotracer, both in the D2 receptor-rich region (striatum) and the D3 receptor-rich region (cerebellum lobes 9 and 10). On the other hand, the occupancies of other test-drugs for D3 receptors were relatively low. In conclusion, we have shown that blonanserin, but not other tested antipsychotics, extensively occupies D3 receptors in vivo in rats. Copyright © 2015 Japanese Pharmacological Society. Production and hosting by Elsevier B.V. All rights reserved.

Three-dimensional Culture of Human Airway Epithelium in Matrigel for Evaluation of Human Rhinovirus C and Bocavirus Infections.

PubMed

Chen, A Xiong; Xie, Guang Cheng; Pan, Dong; DU, Ya Rong; Pang, Li Li; Song, Jing Dong; Duan, Zhao Jun; Hu, Bu Rong

2018-02-01

Newly identified human rhinovirus C (HRV-C) and human bocavirus (HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses. A platform for culturing human airway epithelia in a three-dimensional (3D) pattern using Matrigel as scaffold was developed. The features of 3D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3D cells at designated time points were quantitated by real-time polymerase chain reaction (PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA. Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3D-cultured human airway epithelial (HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3D culture system. Our data provide a preliminary indication that the 3D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV. Copyright © 2018 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

3D stromal tissue equivalent affects intestinal epithelium morphogenesis in vitro.

PubMed

De Gregorio, Vincenza; Imparato, Giorgia; Urciuolo, Francesco; Netti, Paolo A

2018-04-01

Current in vitro models of human intestine commonly fail to mimic the complex intestinal functions and features required for drug development and disease research. Here, we deeply investigate the interaction existing between epithelium and the underneath stroma, and its role in the epithelium morphogenesis. We cultured human intestinal subepithelial myofibroblasts (ISEMFs) in two different 3D configurations: 3D-collagen gel equivalent (3D-CGE) and 3D cell-synthetized stromal equivalent (3D-CSSE). The 3D-CGEs were obtained by means of the traditional collagen-based cell technique and the 3D-CSSE were obtained by bottom-up tissue engineering strategy. The biophysical properties of both 3D models with regard to cell growth and composition (via histological analysis, immunofluorescence, and multiphoton imaging) were assessed. Then, human colorectal adenocarcinoma cell line (CaCo-2) was cultured on both the 3D constructs in order to produce the intestinal model. We identified higher levels of matrix-associated proteins from ISEMFs cultured in 3D-CSSE compared to 3D-CGE. Furthermore, multiphoton investigation revealed differences in the collagen network architecture in both models. At last, the more physiologically relevant stromal environment of the 3D-CSSE drove the CaCo-2 cell differentiation toward the four different type of intestinal epithelial cells (absorptive, mucus-secretory, enteroendocrine, and Paneth) phenotype and promotes, in contrast to the 3D-CGE, the production of the basement membrane. Taken together, these results highlight a fundamental role of the 3D stromal environment in addressing a correct epithelium morphogenesis as well as epithelial-stromal interface establishment. © 2017 Wiley Periodicals, Inc.

Three-Dimensional Engineered High Fidelity Normal Human Lung Tissue-Like Assemblies (TLA) as Targets for Human Respiratory Virus Infections

NASA Technical Reports Server (NTRS)

Goodwin, T. J.; Deatly, A. M.; Suderman, M. T.; Lin, Y.-H.; Chen, W.; Gupta, C. K.; Randolph, V. B.; Udem, S. A.

2003-01-01

Unlike traditional two-dimensional (2D) cell cultures, three-dimensional (3D) tissue-like assemblies (TLA) (Goodwin et aI, 1992, 1993, 2000 and Nickerson et aI. , 2001,2002) offer high organ fidelity with the potential to emulate the infective dynamics of viruses and bacteria in vivo. Thus, utilizing NASA micro gravity Rotating Wall Vessel (RWV) technology, in vitro human broncho-epithelial (HBE) TLAs were engineered to mimic in vivo tissue for study of human respiratory viruses. These 3D HBE TLAs were propagated from a human broncho-tracheal cell line with a mesenchymal component (HBTC) as the foundation matrix and either an adult human broncho-epithelial cell (BEAS-2B) or human neonatal epithelial cell (16HBE140-) as the overlying element. Resulting TLAs share several characteristic features with in vivo human respiratory epithelium including tight junctions, desmosomes and cilia (SEM, TEM). The presence of epithelium and specific lung epithelium markers furthers the contention that these HBE cells differentiate into TLAs paralleling in vivo tissues. A time course of infection of these 3D HBE TLAs with human respiratory syncytial virus (hRSV) wild type A2 strain, indicates that virus replication and virus budding are supported and manifested by increasing virus titer and detection of membrane-bound F and G glycoproteins. Infected 3D HBE TLAs remain intact for up to 12 days compared to infected 2D cultures that are destroyed in 2-3 days. Infected cells show an increased vacuolation and cellular destruction (by transmission electron microscopy) by day 9; whereas, uninfected cells remain robust and morphologically intact. Therefore, the 3D HBE TLAs mimic aspects of human respiratory epithelium providing a unique opportunity to analyze, for the first time, simulated in vivo viral infection independent of host immune response.

Three-dimensional tissue assemblies: novel models for the study of Salmonella enterica serovar Typhimurium pathogenesis

NASA Technical Reports Server (NTRS)

Nickerson, C. A.; Goodwin, T. J.; Terlonge, J.; Ott, C. M.; Buchanan, K. L.; Uicker, W. C.; Emami, K.; LeBlanc, C. L.; Ramamurthy, R.; Clarke, M. S.;

Three-Dimensional Tissue Assemblies: Novel Models for the Study of Salmonella enterica Serovar Typhimurium Pathogenesis

PubMed Central

Nickerson, Cheryl A.; Goodwin, Thomas J.; Terlonge, Jacqueline; Ott, C. Mark; Buchanan, Kent L.; Uicker, William C.; Emami, Kamal; LeBlanc, Carly L.; Ramamurthy, Rajee; Clarke, Mark S.; Vanderburg, Charles R.; Hammond, Timothy; Pierson, Duane L.

2001-01-01

The lack of readily available experimental systems has limited knowledge pertaining to the development of Salmonella-induced gastroenteritis and diarrheal disease in humans. We used a novel low-shear stress cell culture system developed at the National Aeronautics and Space Administration in conjunction with cultivation of three-dimensional (3-D) aggregates of human intestinal tissue to study the infectivity of Salmonella enterica serovar Typhimurium for human intestinal epithelium. Immunohistochemical characterization and microscopic analysis of 3-D aggregates of the human intestinal epithelial cell line Int-407 revealed that the 3-D cells more accurately modeled human in vivo differentiated tissues than did conventional monolayer cultures of the same cells. Results from infectivity studies showed that Salmonella established infection of the 3-D cells in a much different manner than that observed for monolayers. Following the same time course of infection with Salmonella, 3-D Int-407 cells displayed minimal loss of structural integrity compared to that of Int-407 monolayers. Furthermore, Salmonella exhibited significantly lower abilities to adhere to, invade, and induce apoptosis of 3-D Int-407 cells than it did for infected Int-407 monolayers. Analysis of cytokine expression profiles of 3-D Int-407 cells and monolayers following infection with Salmonella revealed significant differences in expression of interleukin 1α (IL-1α), IL-1β, IL-6, IL-1Ra, and tumor necrosis factor alpha mRNAs between the two cultures. In addition, uninfected 3-D Int-407 cells constitutively expressed higher levels of transforming growth factor β1 mRNA and prostaglandin E2 than did uninfected Int-407 monolayers. By more accurately modeling many aspects of human in vivo tissues, the 3-D intestinal cell model generated in this study offers a novel approach for studying microbial infectivity from the perspective of the host-pathogen interaction. PMID:11598087

Three-Dimensional Visualization with Large Data Sets: A Simulation of Spreading Cortical Depression in Human Brain

PubMed Central

Ertürk, Korhan Levent; Şengül, Gökhan

2012-01-01

We developed 3D simulation software of human organs/tissues; we developed a database to store the related data, a data management system to manage the created data, and a metadata system for the management of data. This approach provides two benefits: first of all the developed system does not require to keep the patient's/subject's medical images on the system, providing less memory usage. Besides the system also provides 3D simulation and modification options, which will help clinicians to use necessary tools for visualization and modification operations. The developed system is tested in a case study, in which a 3D human brain model is created and simulated from 2D MRI images of a human brain, and we extended the 3D model to include the spreading cortical depression (SCD) wave front, which is an electrical phoneme that is believed to cause the migraine. PMID:23258956

Bioinspired Three-Dimensional Human Neuromuscular Junction Development in Suspended Hydrogel Arrays.

PubMed

Dixon, Thomas Anthony; Cohen, Eliad; Cairns, Dana M; Rodriguez, Maria; Mathews, Juanita; Jose, Rod R; Kaplan, David L

2018-06-01

The physical connection between motoneurons and skeletal muscle targets is responsible for the creation of neuromuscular junctions (NMJs), which allow electrical signals to be translated to mechanical work. NMJ pathology contributes to the spectrum of neuromuscular, motoneuron, and dystrophic disease. Improving in vitro tools that allow for recapitulation of the physiology of the neuromuscular connection will enable researchers to better understand the development and maturation of NMJs, and will help to decipher mechanisms leading to NMJ degeneration. In this work, we first describe robust differentiation of bungarotoxin-positive human myotubes, as well as a reproducible method for encapsulating and aligning human myoblasts in three-dimensional (3D) suspended culture using bioprinted silk fibroin cantilevers as cell culture supports. Further analysis with coculture of motoneuron-like cells demonstrates feasibility of fully human coculture using two-dimensional and 2.5-dimensional culture methods, with appropriate differentiation of both cell types. Using these coculture differentiation conditions with motoneuron-like cells added to monocultures of 3D suspended human myotubes, we then demonstrate synaptic colocalization in coculture as well as acetylcholine and glutamic acid stimulation of human myocytes. This method represents a unique platform to coculture suspended human myoblast-seeded 3D hydrogels with integrated motoneuron-like cells derived from human induced neural stem cells. The platform described is fully customizable using 3D freeform printing into standard laboratory tissue culture materials, and allows for human myoblast alignment in 3D with precise motoneuron integration into preformed myotubes. The coculture method will ideally be useful in observation and analysis of neurite outgrowth and myogenic differentiation in 3D with quantification of several parameters of muscle innervation and function.

3D Models of Immunotherapy

Cancer.gov

This collaborative grant is developing 3D models of both mouse and human biology to investigate aspects of therapeutic vaccination in order to answer key questions relevant to human cancer immunotherapy.

Cyclic hydrostatic pressure and particles increase synthesis of 1,25-dihydroxyvitamin D3 by human macrophages in vitro.

PubMed

Evans, C E; Mylchreest, S; Mee, A P; Berry, J L; Andrew, J G

2006-01-01

1,25-Dihydroxyvitamin D(3) has a pivotal role in bone resorption and osteoclast activity. As activated macrophages are known to synthesise 1,25-dihydroxyvitamin D(3), this study examined whether pressure modulated its synthesis. Pressure and particles have been shown to increase synthesis of pro-resorptive cytokines and other factors by cultured macrophages. Human peripheral blood macrophages were isolated, cultured and exposed to pressure (similar to that found in the human joint) and/or particles. Synthesis of 1,25-dihydroxyvitamin D(3) by macrophages was assayed using high pressure liquid chromatography and in situ hybridization. Synthesis of 1,25-dihydroxyvitamin D(3) but not 24,25-dihydroxyvitamin D(3) was increased in macrophages under pressure. In situ hybridization demonstrated an increase in 1alpha-hydroxylase expression in response to pressure or particles and simultaneous exposure to both stimuli generated higher expression of 1alpha-hydroxylase. In conclusion, this is the first study to demonstrate that mechanical loading, in the form of pressure, stimulates 1,25-dihydroxyvitamin D(3) synthesis in human macrophages. These findings have implications for the in vivo situation, as they suggest that 1,25-dihydroxyvitamin D(3) could be one factor stimulating osteoclastic bone resorption in pathologies, such as arthritis or implant loosening, where intra-articular or intra-osseous pressure is raised or where wear particles interact with macrophages.

Three-dimensional display technologies

PubMed Central

Geng, Jason

2014-01-01

The physical world around us is three-dimensional (3D), yet traditional display devices can show only two-dimensional (2D) flat images that lack depth (i.e., the third dimension) information. This fundamental restriction greatly limits our ability to perceive and to understand the complexity of real-world objects. Nearly 50% of the capability of the human brain is devoted to processing visual information [Human Anatomy & Physiology (Pearson, 2012)]. Flat images and 2D displays do not harness the brain’s power effectively. With rapid advances in the electronics, optics, laser, and photonics fields, true 3D display technologies are making their way into the marketplace. 3D movies, 3D TV, 3D mobile devices, and 3D games have increasingly demanded true 3D display with no eyeglasses (autostereoscopic). Therefore, it would be very beneficial to readers of this journal to have a systematic review of state-of-the-art 3D display technologies. PMID:25530827

Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo

PubMed Central

Amoroso, Matteo; Lindahl, Anders; Brantsing, Camilla; Rotter, Nicole; Gatenholm, Paul; Kölby, Lars

2017-01-01

Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The development of a process capable of creating a patient-specific cartilage framework would be a major breakthrough. Here, we described methods for creating human cartilage in vivo and quantitatively assessing the proliferative capacity and cartilage-formation ability in mono- and co-cultures of human chondrocytes and human mesenchymal stem cells in a three-dimensional (3D)-bioprinted hydrogel scaffold. The 3D-bioprinted constructs (5 × 5 × 1.2 mm) were produced using nanofibrillated cellulose and alginate in combination with human chondrocytes and human mesenchymal stem cells using a 3D-extrusion bioprinter. Immediately following bioprinting, the constructs were implanted subcutaneously on the back of 48 nude mice and explanted after 30 and 60 days, respectively, for morphological and immunohistochemical examination. During explantation, the constructs were easy to handle, and the majority had retained their macroscopic grid appearance. Constructs consisting of human nasal chondrocytes showed good proliferation ability, with 17.2% of the surface areas covered with proliferating chondrocytes after 60 days. In constructs comprising a mixture of chondrocytes and stem cells, an additional proliferative effect was observed involving chondrocyte production of glycosaminoglycans and type 2 collagen. This clinically highly relevant study revealed 3D bioprinting as a promising technology for the creation of human cartilage. PMID:29236765

Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo.

PubMed

Apelgren, Peter; Amoroso, Matteo; Lindahl, Anders; Brantsing, Camilla; Rotter, Nicole; Gatenholm, Paul; Kölby, Lars

2017-01-01

Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The development of a process capable of creating a patient-specific cartilage framework would be a major breakthrough. Here, we described methods for creating human cartilage in vivo and quantitatively assessing the proliferative capacity and cartilage-formation ability in mono- and co-cultures of human chondrocytes and human mesenchymal stem cells in a three-dimensional (3D)-bioprinted hydrogel scaffold. The 3D-bioprinted constructs (5 × 5 × 1.2 mm) were produced using nanofibrillated cellulose and alginate in combination with human chondrocytes and human mesenchymal stem cells using a 3D-extrusion bioprinter. Immediately following bioprinting, the constructs were implanted subcutaneously on the back of 48 nude mice and explanted after 30 and 60 days, respectively, for morphological and immunohistochemical examination. During explantation, the constructs were easy to handle, and the majority had retained their macroscopic grid appearance. Constructs consisting of human nasal chondrocytes showed good proliferation ability, with 17.2% of the surface areas covered with proliferating chondrocytes after 60 days. In constructs comprising a mixture of chondrocytes and stem cells, an additional proliferative effect was observed involving chondrocyte production of glycosaminoglycans and type 2 collagen. This clinically highly relevant study revealed 3D bioprinting as a promising technology for the creation of human cartilage.

Effect of Curcuma longa on CYP2D6- and CYP3A4-mediated metabolism of dextromethorphan in human liver microsomes and healthy human subjects.

PubMed

Al-Jenoobi, Fahad Ibrahim; Al-Thukair, Areej A; Alam, Mohd Aftab; Abbas, Fawkeya A; Al-Mohizea, Abdullah M; Alkharfy, Khalid M; Al-Suwayeh, Saleh A

2015-03-01

Effect of Curcuma longa rhizome powder and its ethanolic extract on CYP2D6 and CYP3A4 metabolic activity was investigated in vitro using human liver microsomes and clinically in healthy human subjects. Dextromethorphan (DEX) was used as common probe for CYP2D6 and CYP3A4 enzymes. Metabolic activity of CYP2D6 and CYP3A4 was evaluated through in vitro study; where microsomes were incubated with NADPH in presence and absence of Curcuma extract. In clinical study phase-I, six healthy human subjects received a single dose (30 mg) of DEX syrup, and in phase-II DEX syrup was administered with Curcuma powder. The enzyme CYP2D6 and CYP3A4 mediated O- and N-demethylation of dextromethorphan into dextrorphan (DOR) and 3-methoxymorphinan (3-MM), respectively. Curcuma extract significantly inhibited the formation of DOR and 3-MM, in a dose-dependent and linear fashion. The 100 μg/ml dose of curcuma extract produced highest inhibition, which was about 70 % for DOR and 80 % for 3-MM. Curcuma significantly increases the urine metabolic ratio of DEX/DOR but the change in DEX/3-MM ratio was statistically insignificant. Present findings suggested that curcuma significantly inhibits the activity of CYP2D6 in in vitro as well as in vivo; which indicates that curcuma has potential to interact with CYP2D6 substrates.

3D Bioprinting Human Chondrocytes with Nanocellulose-Alginate Bioink for Cartilage Tissue Engineering Applications.

PubMed

Markstedt, Kajsa; Mantas, Athanasios; Tournier, Ivan; Martínez Ávila, Héctor; Hägg, Daniel; Gatenholm, Paul

2015-05-11

The introduction of 3D bioprinting is expected to revolutionize the field of tissue engineering and regenerative medicine. The 3D bioprinter is able to dispense materials while moving in X, Y, and Z directions, which enables the engineering of complex structures from the bottom up. In this study, a bioink that combines the outstanding shear thinning properties of nanofibrillated cellulose (NFC) with the fast cross-linking ability of alginate was formulated for the 3D bioprinting of living soft tissue with cells. Printability was evaluated with concern to printer parameters and shape fidelity. The shear thinning behavior of the tested bioinks enabled printing of both 2D gridlike structures as well as 3D constructs. Furthermore, anatomically shaped cartilage structures, such as a human ear and sheep meniscus, were 3D printed using MRI and CT images as blueprints. Human chondrocytes bioprinted in the noncytotoxic, nanocellulose-based bioink exhibited a cell viability of 73% and 86% after 1 and 7 days of 3D culture, respectively. On the basis of these results, we can conclude that the nanocellulose-based bioink is a suitable hydrogel for 3D bioprinting with living cells. This study demonstrates the potential use of nanocellulose for 3D bioprinting of living tissues and organs.

Characterization of Phenotypic and Transcriptional Differences in Human Pluripotent Stem Cells under 2D and 3D Culture Conditions.

PubMed

Kamei, Ken-Ichiro; Koyama, Yoshie; Tokunaga, Yumie; Mashimo, Yasumasa; Yoshioka, Momoko; Fockenberg, Christopher; Mosbergen, Rowland; Korn, Othmar; Wells, Christine; Chen, Yong

2016-11-01

Human pluripotent stem cells hold great promise for applications in drug discovery and regenerative medicine. Microfluidic technology is a promising approach for creating artificial microenvironments; however, although a proper 3D microenvironment is required to achieve robust control of cellular phenotypes, most current microfluidic devices provide only 2D cell culture and do not allow tuning of physical and chemical environmental cues simultaneously. Here, the authors report a 3D cellular microenvironment plate (3D-CEP), which consists of a microfluidic device filled with thermoresponsive poly(N-isopropylacrylamide)-β-poly(ethylene glycol) hydrogel (HG), which enables systematic tuning of both chemical and physical environmental cues as well as in situ cell monitoring. The authors show that H9 human embryonic stem cells (hESCs) and 253G1 human induced pluripotent stem cells in the HG/3D-CEP system maintain their pluripotent marker expression under HG/3D-CEP self-renewing conditions. Additionally, global gene expression analyses are used to elucidate small variations among different test environments. Interestingly, the authors find that treatment of H9 hESCs under HG/3D-CEP self-renewing conditions results in initiation of entry into the neural differentiation process by induction of PAX3 and OTX1 expression. The authors believe that this HG/3D-CEP system will serve as a versatile platform for developing targeted functional cell lines and facilitate advances in drug screening and regenerative medicine. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Real-time 3D human capture system for mixed-reality art and entertainment.

PubMed

Nguyen, Ta Huynh Duy; Qui, Tran Cong Thien; Xu, Ke; Cheok, Adrian David; Teo, Sze Lee; Zhou, ZhiYing; Mallawaarachchi, Asitha; Lee, Shang Ping; Liu, Wei; Teo, Hui Siang; Thang, Le Nam; Li, Yu; Kato, Hirokazu

2005-01-01

A real-time system for capturing humans in 3D and placing them into a mixed reality environment is presented in this paper. The subject is captured by nine cameras surrounding her. Looking through a head-mounted-display with a camera in front pointing at a marker, the user can see the 3D image of this subject overlaid onto a mixed reality scene. The 3D images of the subject viewed from this viewpoint are constructed using a robust and fast shape-from-silhouette algorithm. The paper also presents several techniques to produce good quality and speed up the whole system. The frame rate of our system is around 25 fps using only standard Intel processor-based personal computers. Besides a remote live 3D conferencing and collaborating system, we also describe an application of the system in art and entertainment, named Magic Land, which is a mixed reality environment where captured avatars of human and 3D computer generated virtual animations can form an interactive story and play with each other. This system demonstrates many technologies in human computer interaction: mixed reality, tangible interaction, and 3D communication. The result of the user study not only emphasizes the benefits, but also addresses some issues of these technologies.

A 3D human neural cell culture system for modeling Alzheimer’s disease

PubMed Central

Kim, Young Hye; Choi, Se Hoon; D’Avanzo, Carla; Hebisch, Matthias; Sliwinski, Christopher; Bylykbashi, Enjana; Washicosky, Kevin J.; Klee, Justin B.; Brüstle, Oliver; Tanzi, Rudolph E.; Kim, Doo Yeon

2015-01-01

Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer’s disease (AD) because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel three-dimensional (3D) culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of β-amyloid and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix, and the analysis of AD pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of β-amyloid is observed after 6-weeks of differentiation followed by robust tau pathology after 10–14 weeks. PMID:26068894

3-D PARTICLE TRANSPORT WITHIN THE HUMAN UPPER RESPIRATORY TRACT

EPA Science Inventory

In this study trajectories of inhaled particulate matter (PM) were simulated within a three-dimensional (3-D) computer model of the human upper respiratory tract (URT). The airways were described by computer-reconstructed images of a silicone rubber cast of the human head, throat...

The three-dimensional microanatomy of the rabbit and human cornea. A chemical and mechanical microdissection-SEM approach

PubMed Central

OJEDA, JOSÉ L.; VENTOSA, JUAN A.; PIEDRA, SONSOLES

2001-01-01

The three-dimensional (3D) microanatomy of the cornea is the major determinant of its optical and mechanical properties. Scanning electron microscopy (SEM) is the most commonly used method to obtain information on the overall 3D microanatomy of organs. However, SEM has not been successful in revealing the 3D microanatomy of the cornea, because the interior of the cornea is too compact to be explored by the electron beam. In this study, the 3D organisation of the cells and extracellular materials of human and rabbit corneas was examined after exposure by HCl and NaOH digestion, and by microdissection by the adhesive tape method. In the cornea of both species, all epithelial cells exhibited microplicae regardless of their location. This raises doubts about the tear film-holding role assigned to the microplicae of the superficial cells. Human and rabbit corneas differed in the collagen fibre patterns of the epithelial basement membranes. The 3D organisation of the stromal lamellae was similar in both species. In humans and rabbits, the keratocytes showed similar 3D features. However, the surface of human keratocytes located near Descemet's membrane exhibited small fenestrations that were not present in the rabbit keratocytes. The pattern of keratocyte innervation by the stromal neural plexus and 3D keratocyte microanatomy confirms that keratocytes form a large intercommunicating network within the corneal stroma. Two morphologically discrete subpopulations of keratocytes located at different stromal levels were identified in both human and rabbit corneas, suggesting that keratocytes are not functionally homogeneous. In addition, the density of the stromal neural plexus appeared to be greater in rabbits than in humans. Clear differences between human and rabbit corneas were observed in the collagen arrangement in Descemet's membrane, which may reflect their different biomechanical requirements. PMID:11760887

Using subject-specific three-dimensional (3D) anthropometry data in digital human modelling: case study in hand motion simulation.

PubMed

Tsao, Liuxing; Ma, Liang

2016-11-01

Digital human modelling enables ergonomists and designers to consider ergonomic concerns and design alternatives in a timely and cost-efficient manner in the early stages of design. However, the reliability of the simulation could be limited due to the percentile-based approach used in constructing the digital human model. To enhance the accuracy of the size and shape of the models, we proposed a framework to generate digital human models using three-dimensional (3D) anthropometric data. The 3D scan data from specific subjects' hands were segmented based on the estimated centres of rotation. The segments were then driven in forward kinematics to perform several functional postures. The constructed hand models were then verified, thereby validating the feasibility of the framework. The proposed framework helps generate accurate subject-specific digital human models, which can be utilised to guide product design and workspace arrangement. Practitioner Summary: Subject-specific digital human models can be constructed under the proposed framework based on three-dimensional (3D) anthropometry. This approach enables more reliable digital human simulation to guide product design and workspace arrangement.

Human Pol ζ purified with accessory subunits is active in translesion DNA synthesis and complements Pol η in cisplatin bypass

PubMed Central

Lee, Young-Sam; Gregory, Mark T.; Yang, Wei

2014-01-01

DNA polymerase ζ (Pol ζ) is a eukaryotic B-family DNA polymerase that specializes in translesion synthesis and is essential for normal embryogenesis. At a minimum, Pol ζ consists of a catalytic subunit Rev3 and an accessory subunit Rev7. Mammalian Rev3 contains >3,000 residues and is twice as large as the yeast homolog. To date, no vertebrate Pol ζ has been purified for biochemical characterization. Here we report purification of a series of human Rev3 deletion constructs expressed in HEK293 cells and identification of a minimally catalytically active human Pol ζ variant. With a tagged form of an active Pol ζ variant, we isolated two additional accessory subunits of human Pol ζ, PolD2 and PolD3. The purified four-subunit Pol ζ4 (Rev3–Rev7–PolD2–PolD3) is much more efficient and more processive at bypassing a 1,2-intrastrand d(GpG)-cisplatin cross-link than the two-subunit Pol ζ2 (Rev3–Rev7). We show that complete bypass of cisplatin lesions requires Pol η to insert dCTP opposite the 3′ guanine and Pol ζ4 to extend the primers. PMID:24449906

Human Pol ζ purified with accessory subunits is active in translesion DNA synthesis and complements Pol η in cisplatin bypass.

PubMed

Lee, Young-Sam; Gregory, Mark T; Yang, Wei

2014-02-25

DNA polymerase ζ (Pol ζ) is a eukaryotic B-family DNA polymerase that specializes in translesion synthesis and is essential for normal embryogenesis. At a minimum, Pol ζ consists of a catalytic subunit Rev3 and an accessory subunit Rev7. Mammalian Rev3 contains >3,000 residues and is twice as large as the yeast homolog. To date, no vertebrate Pol ζ has been purified for biochemical characterization. Here we report purification of a series of human Rev3 deletion constructs expressed in HEK293 cells and identification of a minimally catalytically active human Pol ζ variant. With a tagged form of an active Pol ζ variant, we isolated two additional accessory subunits of human Pol ζ, PolD2 and PolD3. The purified four-subunit Pol ζ4 (Rev3-Rev7-PolD2-PolD3) is much more efficient and more processive at bypassing a 1,2-intrastrand d(GpG)-cisplatin cross-link than the two-subunit Pol ζ2 (Rev3-Rev7). We show that complete bypass of cisplatin lesions requires Pol η to insert dCTP opposite the 3' guanine and Pol ζ4 to extend the primers.

gp140, the EBV/C3d receptor (CR2) of human B lymphocytes, is involved in cell-free phosphorylation of p120, a nuclear ribonucleoprotein.

PubMed

Delcayre, A X; Fiandino, A; Barel, M; Frade, R

1987-12-01

gp140, the EB/C3d receptor (EBV/C3dR; CR2), is a membrane site involved in human B cell regulation. Cross-linking of this receptor on the cell surface by its specific ligands led to the enhancement of B cell proliferation in synergy with T cell factors. In vitro activation of human peripheral B lymphocytes by cross-linking membrane immunoglobulins with anti-mu antibody induced EBV/C3dR phosphorylation. These studies were pursued by analyzing cell-free phosphorylation of EBV/C3dR isolated from Raji cell fractions, and immobilized on OKB7, a monoclonal anti-EBV/C3dR antibody. Three EBV/C3dR-related antigens which could be cell-free phosphorylated were detected: gp140, the EBV/C3dR, p130 and p120. gp140, the mature form of EBV/C3dR, was isolated from plasma membrane and from purified nuclei. p130 was identified as an intracellular intermediate of EBV/C3dR glycosylation, localized in low-density microsomes. Phosphoamino acid analysis of EBV/C3dR allowed the detection of phosphotyrosine and phosphoserine residues. These data suggest that EBV/C3dR could carry an autophosphorylation activity and could be associated to serine kinases. Using polyclonal anti-p120 antibody and anti-120 kDa nuclear ribonucleoprotein monoclonal antibody (mAb), p120 was identified as a nuclear ribonucleoprotein antigenically not related to EBV/C3dR. Detection of p120 on EBV/C3dR, immobilized on OKB7, was due to interactions between both antigens, instead of anti-EBV/C3dR mAb cross-reactivity with p120. Cell-free phosphorylation of p120 was under the control of EBV/C3dR. However, it is not yet established whether other nuclear or membrane components were involved in the control of p120 cell-free phosphorylation by EBV/C3dR. From the data presented herein, we propose that phosphorylation of a 120-kDa nuclear ribonucleoprotein by EBV/C3dR-associated kinases could represent a crucial step in in vivo regulation of human B cell activation.

Methodologies for Development of Patient Specific Bone Models from Human Body CT Scans

NASA Astrophysics Data System (ADS)

Chougule, Vikas Narayan; Mulay, Arati Vinayak; Ahuja, Bharatkumar Bhagatraj

2016-06-01

This work deals with development of algorithm for physical replication of patient specific human bone and construction of corresponding implants/inserts RP models by using Reverse Engineering approach from non-invasive medical images for surgical purpose. In medical field, the volumetric data i.e. voxel and triangular facet based models are primarily used for bio-modelling and visualization, which requires huge memory space. On the other side, recent advances in Computer Aided Design (CAD) technology provides additional facilities/functions for design, prototyping and manufacturing of any object having freeform surfaces based on boundary representation techniques. This work presents a process to physical replication of 3D rapid prototyping (RP) physical models of human bone from various CAD modeling techniques developed by using 3D point cloud data which is obtained from non-invasive CT/MRI scans in DICOM 3.0 format. This point cloud data is used for construction of 3D CAD model by fitting B-spline curves through these points and then fitting surface between these curve networks by using swept blend techniques. This process also can be achieved by generating the triangular mesh directly from 3D point cloud data without developing any surface model using any commercial CAD software. The generated STL file from 3D point cloud data is used as a basic input for RP process. The Delaunay tetrahedralization approach is used to process the 3D point cloud data to obtain STL file. CT scan data of Metacarpus (human bone) is used as the case study for the generation of the 3D RP model. A 3D physical model of the human bone is generated on rapid prototyping machine and its virtual reality model is presented for visualization. The generated CAD model by different techniques is compared for the accuracy and reliability. The results of this research work are assessed for clinical reliability in replication of human bone in medical field.

Three-dimensional surface anthropometry: Applications to the human body

NASA Astrophysics Data System (ADS)

Jones, Peter R. M.; Rioux, Marc

1997-09-01

Anthropometry is the study of the measurement of the human body. By tradition this has been carried out taking the measurements from body surface landmarks, such as circumferences and breadths, using simple instruments like tape measures and calipers. Three-dimensional (3D) surface anthropometry enables us to extend the study to 3D geometry and morphology of mainly external human body tissues. It includes the acquisition, indexing, transmission, archiving, retrieval, interrogation and analysis of body size, shape, and surface together with their variability throughout growth and development to adulthood. While 3D surface anthropometry surveying is relatively new, anthropometric surveying using traditional tools, such as calipers and tape measures, is not. Recorded studies of the human form date back to ancient times. Since at least the 17th century 1 investigators have made attempts to measure the human body for physical properties such as weight, size, and centre of mass. Martin documented 'standard' body measurement methods in a handbook in 1928. 2 This paper reviews the past and current literature devoted to the applications of 3D anthropometry because true 3D scanning of the complete human body is fast becoming a reality. We attempt to take readers through different forms of technology which deal with simple forms of projected light to the more complex advanced forms of laser and video technology giving low and/or high resolution 3D data. Information is also given about image capture of size and shape of the whole as well as most component parts of the human body. In particular, the review describes with explanations a multitude of applications, for example, medical, product design, human engineering, anthropometry and ergonomics etc.

Human Milk Proresolving Mediators Stimulate Resolution of Acute Inflammation

PubMed Central

Dalli, Jesmond; Serhan, Charles N

2015-01-01

Human milk contains nutrients and bioactive products relevant to infant development and immunological protection. Here, we investigated the pro-resolving properties of milk using human milk lipid mediator isolates (HLMI) and determined their impact on resolution programs in vivo and with human macrophages. HLMI reduced maximum neutrophil numbers (14.6±1.2×106 to 11.0±1.0×106 cells/exudate) and shortened the resolution interval (Ri; 50% neutrophil reduction) 54% compared to peritonitis. Using rigorous liquid-chromatography tandem-mass spectrometry (LC-MS-MS)-based lipid mediator (LM) metabololipidomics, we demonstrated that human milk possesses a proresolving LM-SPM signature profile, containing specialized proresolving mediators (SPM; e.g. resolvins, protectins, maresins and lipoxins) at bioactive levels (pico-nanomolar concentrations) that enhanced human macrophage efferocytosis and bacterial containment. SPM identified in human milk included D-series resolvins, (e.g. Resolvin (Rv) D1, RvD2, RvD3, AT-RvD3 and RvD4), Protectin (PD)1, Maresin (MaR)1, E-series resolvins (e.g. RvE1, RvE2 and RvE3) and lipoxins (LXA4 and LXB4). Of the SPM identified in human milk, RvD2 and MaR1 (50 ng/mouse) individually shortened Ri ~75%. Milk from mastitis gave higher LTB4 and prostanoids and lower SPM levels. Taken together, these findings provide evidence that human milk has pro-resolving actions via comprehensive LM-SPM profiling, describing a potentially novel mechanism in maternal-infant biochemical imprinting. PMID:26462421

CCQM-K132: low-polarity analytes in a biological matrix: vitamin D metabolites in human serum

NASA Astrophysics Data System (ADS)

Wise, Stephen A.; Tai, Susan S.-C.; Duewer, David L.; Bedner, Mary; Camara, Johanna E.; Lippa, Katrice A.; Qinde, Liu; Kang, Dukjin; Kim, Byungjoo; Quan, Can; Shi, Lianhua; Nammoonnoy, Jintana; Vamathevan, Veronica; Ceyhan Gören, Ahmet; Bilsel, Gökhan; Yilmaz, Hasibe

2017-01-01

Vitamin D is a fat-soluble vitamin that occurs primarily in two forms, vitamin D2 and vitamin D3. Vitamin D3 is produced naturally when skin is exposed to UV radiation, is naturally-occurring in foods (generally of animal origin), and is fortified in some foods and dietary supplements. Vitamin D2 occurs in food (generally plant sources) and until recently was the form most often used in dietary supplements. Vitamin D is metabolized in the body to produce several closely related, hydroxylated species (metabolites), with 25-hydroxyvitamin D3 [25(OH)D3] and 25-hydroxyvitamin D2 [25(OH)D2] as the most common metabolites measured in human serum. Concentrations of total vitamin D in human serum, calculated as the sum of 25(OH)D2 and 25(OH)D3, are typically in the 16 ng/g to 30 ng/g (40 nmol/L to 75 nmol/L) range, with 25(OH)D3 usually accounting for more than 90 % of the total. An epimer of 25(OH)D3, 3-epi-25(OH)D3, can be present at levels up to 10 % of 25(OH)D3 concentration. Seven National Metrology Institutions participated in the Track C Key Comparison CCQM-K132 low-polarity analytes in a biological matrix: vitamin D metabolites in human serum. Participants were requested to evaluate the mass fractions, expressed in ng/g, of 25(OH)D3, 25(OH)D2, and 3-epi-25(OH)D3 in two human serum materials, termed Serum Pool I and Serum Pool II. Due to the known low levels of 3-epi-25(OH)D3 in both materials and the very low level of 25(OH)D2 in Serum Pool I, the study protocol stated that key comparison reference values (KCRVs) would be assigned only to 25(OH)D3 in both materials and 25(OH)D2 in Serum Pool II. Results for 3-epi-25(OH)D3 were requested to evaluate the separation technologies employed; 3-epi-25(OH)D3 needs to be chromatographically separated from 25(OH)D3 for proper quantification of 25(OH)D3. Results for 25(OH)D2 in Serum Pool I were requested to explore measurement performance at its low level. All participants used isotope dilution liquid chromatography with tandem mass spectrometry detection (ID LC-MS/MS) for the measurement of the vitamin D metabolites. Successful participation in CCQM K132 demonstrates capabilities in analysis of low molecular mass (100 g/mol to 500 g/mol) and low-polarity (nonpolar, pKow < -2) analytes at the 1 ng/g to 500 ng/g mass fraction range in complex biological matrixes with core competencies for sample preparation and analysis using ID LC-MS/MS. This study extends the mass fraction capability range to 105 to 106 times lower than that demonstrated in previous CCQM Key Comparisons for cholesterol in serum, another nonpolar clinical analyte. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Ex vivo 2D and 3D HSV-2 infection model using human normal vaginal epithelial cells.

PubMed

Zhu, Yaqi; Yang, Yan; Guo, Juanjuan; Dai, Ying; Ye, Lina; Qiu, Jianbin; Zeng, Zhihong; Wu, Xiaoting; Xing, Yanmei; Long, Xiang; Wu, Xufeng; Ye, Lin; Wang, Shubin; Li, Hui

2017-02-28

Herpes simplex virus type 2 (HSV-2) infects human genital mucosa and establishes life-long latent infection. It is unmet need to establish a human cell-based microphysiological system for virus biology and anti-viral drug discovery. One of barriers is lacking of culture system of normal epithelial cells in vitro over decades. In this study, we established human normal vaginal epithelial cell (HNVEC) culture using co-culture system. HNVEC cells were then propagated rapidly and stably in a defined culture condition. HNVEC cells exhibited a normal diploid karyotype and formed the well-defined and polarized spheres in matrigel three-dimension (3D) culture, while malignant cells (HeLa) formed disorganized and nonpolar solid spheres. HNVEC cells had a normal cellular response to DNA damage and had no transforming property using soft agar assays. HNVEC expressed epithelial marker cytokeratin 14 (CK14) and p63, but not cytokeratin 18 (CK18). Next, we reconstructed HNVEC-derived 3D vaginal epithelium using air-liquid interface (ALI) culture. This 3D vaginal epithelium has the basal and apical layers with expression of epithelial markers as its originated human vaginal tissue. Finally, we established an HSV-2 infection model based on the reconstructed 3D vaginal epithelium. After inoculation of HSV-2 (G strain) at apical layer of the reconstructed 3D vaginal epithelium, we observed obvious pathological effects gradually spreading from the apical layer to basal layer with expression of a viral protein. Thus, we established an ex vivo 2D and 3D HSV-2 infection model that can be used for HSV-2 virology and anti-viral drug discovery.

CJ-1639: A Potent and Highly Selective Dopamine D3 Receptor Full Agonist.

PubMed

Chen, Jianyong; Collins, Gregory T; Levant, Beth; Woods, James; Deschamps, Jeffrey R; Wang, Shaomeng

2011-08-11

We have identified several ligands with high binding affinities to the dopamine D3 receptor and excellent selectivity over the D2 and D1 receptors. CJ-1639 (17) binds to the D3 receptor with a K(i) value of 0.50 nM and displays a selectivity of >5,000 times over D2 and D1 receptors in binding assays using dopamine receptors expressed in the native rat brain tissues. CJ-1639 binds to human D3 receptor with a K(i) value of 3.61 nM and displays over >1000-fold selectivity over human D1 and D2 receptors. CJ-1639 is active at 0.01 mg/kg at the dopamine D3 receptor in the rat and only starts to show a modest D2 activity at doses as high as 10 mg/kg. CJ-1639 is the most potent and selective D3 full agonist reported to date.

Development of a stereoscopic three-dimensional drawing application

NASA Astrophysics Data System (ADS)

Carver, Donald E.; McAllister, David F.

1991-08-01

With recent advances in 3-D technology, computer users have the opportunity to work within a natural 3-D environment; a flat panel LCD computer display of this type, the DTI-100M made by Dimension Technologies, Inc., recently went on the market. In a joint venture between DTI and NCSU, an object-oriented 3-D drawing application, 3-D Draw, was developed to address some issues of human interface design for interactive stereo drawing applications. The focus of this paper is to determine some of the procedures a user would naturally expect to follow while working within a true 3-D environment. The paper discusses (1) the interface between the Macintosh II and DTI-100M during implementation of 3-D Draw, including stereo cursor development and presentation of current 2-D systems, with an additional `depth'' parameter, in the 3-D world, (2) problems in general for human interface into the 3-D environment, and (3) necessary functions and/or problems in developing future stereoscopic 3-D operating systems/tools.

3D FaceCam: a fast and accurate 3D facial imaging device for biometrics applications

NASA Astrophysics Data System (ADS)

Geng, Jason; Zhuang, Ping; May, Patrick; Yi, Steven; Tunnell, David

2004-08-01

Human faces are fundamentally three-dimensional (3D) objects, and each face has its unique 3D geometric profile. The 3D geometric features of a human face can be used, together with its 2D texture, for rapid and accurate face recognition purposes. Due to the lack of low-cost and robust 3D sensors and effective 3D facial recognition (FR) algorithms, almost all existing FR systems use 2D face images. Genex has developed 3D solutions that overcome the inherent problems in 2D while also addressing limitations in other 3D alternatives. One important aspect of our solution is a unique 3D camera (the 3D FaceCam) that combines multiple imaging sensors within a single compact device to provide instantaneous, ear-to-ear coverage of a human face. This 3D camera uses three high-resolution CCD sensors and a color encoded pattern projection system. The RGB color information from each pixel is used to compute the range data and generate an accurate 3D surface map. The imaging system uses no moving parts and combines multiple 3D views to provide detailed and complete 3D coverage of the entire face. Images are captured within a fraction of a second and full-frame 3D data is produced within a few seconds. This described method provides much better data coverage and accuracy in feature areas with sharp features or details (such as the nose and eyes). Using this 3D data, we have been able to demonstrate that a 3D approach can significantly improve the performance of facial recognition. We have conducted tests in which we have varied the lighting conditions and angle of image acquisition in the "field." These tests have shown that the matching results are significantly improved when enrolling a 3D image rather than a single 2D image. With its 3D solutions, Genex is working toward unlocking the promise of powerful 3D FR and transferring FR from a lab technology into a real-world biometric solution.

Human stem cell based corneal tissue mimicking structures using laser-assisted 3D bioprinting and functional bioinks.

PubMed

Sorkio, Anni; Koch, Lothar; Koivusalo, Laura; Deiwick, Andrea; Miettinen, Susanna; Chichkov, Boris; Skottman, Heli

2018-07-01

There is a high demand for developing methods to produce more native-like 3D corneal structures. In the present study, we produced 3D cornea-mimicking tissues using human stem cells and laser-assisted bioprinting (LaBP). Human embryonic stem cell derived limbal epithelial stem cells (hESC-LESC) were used as a cell source for printing epithelium-mimicking structures, whereas human adipose tissue derived stem cells (hASCs) were used for constructing layered stroma-mimicking structures. The development and optimization of functional bioinks was a crucial step towards successful bioprinting of 3D corneal structures. Recombinant human laminin and human sourced collagen I served as the bases for the functional bioinks. We used two previously established LaBP setups based on laser induced forward transfer, with different laser wavelengths and appropriate absorption layers. We bioprinted three types of corneal structures: stratified corneal epithelium using hESC-LESCs, lamellar corneal stroma using alternating acellular layers of bioink and layers with hASCs, and finally structures with both a stromal and epithelial part. The printed constructs were evaluated for their microstructure, cell viability and proliferation, and key protein expression (Ki67, p63α, p40, CK3, CK15, collagen type I, VWF). The 3D printed stromal constructs were also implanted into porcine corneal organ cultures. Both cell types maintained good viability after printing. Laser-printed hESC-LESCs showed epithelial cell morphology, expression of Ki67 proliferation marker and co-expression of corneal progenitor markers p63α and p40. Importantly, the printed hESC-LESCs formed a stratified epithelium with apical expression of CK3 and basal expression of the progenitor markers. The structure of the 3D bioprinted stroma demonstrated that the hASCs had organized horizontally as in the native corneal stroma and showed positive labeling for collagen I. After 7 days in porcine organ cultures, the 3D bioprinted stromal structures attached to the host tissue with signs of hASCs migration from the printed structure. This is the first study to demonstrate the feasibility of 3D LaBP for corneal applications using human stem cells and successful fabrication of layered 3D bioprinted tissues mimicking the structure of the native corneal tissue. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

Comparative analysis of respiratory motion tracking using Microsoft Kinect v2 sensor.

PubMed

Silverstein, Evan; Snyder, Michael

2018-05-01

To present and evaluate a straightforward implementation of a marker-less, respiratory motion-tracking process utilizing Kinect v2 camera as a gating tool during 4DCT or during radiotherapy treatments. Utilizing the depth sensor on the Kinect as well as author written C# code, respiratory motion of a subject was tracked by recording depth values obtained at user selected points on the subject, with each point representing one pixel on the depth image. As a patient breathes, specific anatomical points on the chest/abdomen will move slightly within the depth image across pixels. By tracking how depth values change for a specific pixel, instead of how the anatomical point moves throughout the image, a respiratory trace can be obtained based on changing depth values of the selected pixel. Tracking these values was implemented via marker-less setup. Varian's RPM system and the Anzai belt system were used in tandem with the Kinect to compare respiratory traces obtained by each using two different subjects. Analysis of the depth information from the Kinect for purposes of phase- and amplitude-based binning correlated well with the RPM and Anzai systems. Interquartile Range (IQR) values were obtained comparing times correlated with specific amplitude and phase percentages against each product. The IQR time spans indicated the Kinect would measure specific percentage values within 0.077 s for Subject 1 and 0.164 s for Subject 2 when compared to values obtained with RPM or Anzai. For 4DCT scans, these times correlate to less than 1 mm of couch movement and would create an offset of 1/2 an acquired slice. By tracking depth values of user selected pixels within the depth image, rather than tracking specific anatomical locations, respiratory motion can be tracked and visualized utilizing the Kinect with results comparable to that of the Varian RPM and Anzai belt. © 2018 The Authors. Journal of Applied Clinical Medical Physics published by Wiley Periodicals, Inc. on behalf of American Association of Physicists in Medicine.

Endoplasmic reticulum-associated degradation of the mouse PC1/3-N222D hypomorph and human PCSK1 mutations contributes to obesity.

PubMed

Stijnen, P; Brouwers, B; Dirkx, E; Ramos-Molina, B; Van Lommel, L; Schuit, F; Thorrez, L; Declercq, J; Creemers, J W M

2016-06-01

The proprotein convertase 1/3 (PC1/3), encoded by proprotein convertase subtilisin/kexin type 1 (PCSK1), cleaves and hence activates several orexigenic and anorexigenic proproteins. Congenital inactivation of PCSK1 leads to obesity in human but not in mice. However, a mouse model harboring the hypomorphic mutation N222D is obese. It is not clear why the mouse models differ in phenotype. Gene expression analysis was performed with pancreatic islets from Pcsk1(N222D/N222D) mice. Subsequently, biosynthesis, maturation, degradation and activity were studied in islets, pituitary, hypothalamus and cell lines. Coimmunoprecipitation of PC1/3-N222D and human PC1/3 variants associated with obesity with the endoplasmic reticulum (ER) chaperone BiP was studied in cell lines. Gene expression analysis of islets of Pcsk1(N222D/N222D) mice showed enrichment of gene sets related to the proteasome and the unfolded protein response. Steady-state levels of PC1/3-N222D and in particular the carboxy-terminally processed form were strongly reduced in islets, pituitary and hypothalamus. However, impairment of substrate cleavage was tissue dependent. Proinsulin processing was drastically reduced, while processing of proopiomelanocortin (POMC) to adrenocorticotropic hormone (ACTH) in pituitary was only mildly impaired. Growth hormone expression and IGF-1 levels were normal, indicating near-normal processing of hypothalamic proGHRH. PC1/3-N222D binds to BiP and is rapidly degraded by the proteasome. Analysis of human PC1/3 obesity-associated mutations showed increased binding to BiP and prolonged intracellular retention for all investigated mutations, in particular for PC1/3-T175M, PC1/3-G226R and PC1/3-G593R. This study demonstrates that the hypomorphic mutation in Pcsk1(N222D) mice has an effect on catalytic activity in pancreatic islets, pituitary and hypothalamus. Reduced substrate processing activity in Pcsk1(N222D/N222D) mice is due to enhanced degradation in addition to reduced catalytic activity of the mutant. PC1/3-N222D binds to BiP, suggesting impaired folding and reduced stability. Enhanced BiP binding is also observed in several human obesity-associated PC1/3 variants, suggesting a common mechanism.

Human3.6M: Large Scale Datasets and Predictive Methods for 3D Human Sensing in Natural Environments.

PubMed

Ionescu, Catalin; Papava, Dragos; Olaru, Vlad; Sminchisescu, Cristian

2014-07-01

We introduce a new dataset, Human3.6M, of 3.6 Million accurate 3D Human poses, acquired by recording the performance of 5 female and 6 male subjects, under 4 different viewpoints, for training realistic human sensing systems and for evaluating the next generation of human pose estimation models and algorithms. Besides increasing the size of the datasets in the current state-of-the-art by several orders of magnitude, we also aim to complement such datasets with a diverse set of motions and poses encountered as part of typical human activities (taking photos, talking on the phone, posing, greeting, eating, etc.), with additional synchronized image, human motion capture, and time of flight (depth) data, and with accurate 3D body scans of all the subject actors involved. We also provide controlled mixed reality evaluation scenarios where 3D human models are animated using motion capture and inserted using correct 3D geometry, in complex real environments, viewed with moving cameras, and under occlusion. Finally, we provide a set of large-scale statistical models and detailed evaluation baselines for the dataset illustrating its diversity and the scope for improvement by future work in the research community. Our experiments show that our best large-scale model can leverage our full training set to obtain a 20% improvement in performance compared to a training set of the scale of the largest existing public dataset for this problem. Yet the potential for improvement by leveraging higher capacity, more complex models with our large dataset, is substantially vaster and should stimulate future research. The dataset together with code for the associated large-scale learning models, features, visualization tools, as well as the evaluation server, is available online at http://vision.imar.ro/human3.6m.

Effect of 1,25-dihydroxyvitamin D3 on human keratinocytes grown under different culture conditions.

PubMed

McLane, J A; Katz, M; Abdelkader, N

1990-04-01

1,25-Dihydroxyvitamin D3 (1,25-(OH)2-D3) is known to decrease the proliferation and increase the differentiation of different cell types including human keratinocytes. The growth and differentiation of keratinocytes in the presence of 1,25-(OH)2-D3 using serum-free media formulations has been described previously. This investigation extends these studies to describe various culture conditions with human foreskin keratinocytes to determine the optimal antiproliferative activity of 1,25-(OH)2-D3. Keratinocytes were plated onto tissue culture dishes using one of three basic serum-free media protocols; a) with no feeder layer in keratinocyte growth medium (KGM); b) onto mitomycin C-treated 3T3 mouse embryo fibroblasts; or c) onto mitomycin C-treated dermal human fibroblasts. The last two protocols utilized Dulbecco's modified Eagle's Medium (DMEM) supplemented with growth factors. Keratinocyte cell growth was greatest in the KGM medium. Although the growth of keratinocytes on either feeder layer was similar, there were differences in the ability of the cells to form envelopes in the presence of 1,25-(OH)2-D3. The addition of hydrocortisone and cholera toxin to the medium also affected the response of the keratinocytes to 1,25-(OH)2-D3. The antiproliferative effect of 1,25-(OH)2-D3 was not altered by varying the extracellular calcium levels from 0.25 to 3 mM. The antiproliferative activity of 1,25-(OH)2-D3 is attenuated in cells at low density. Our results suggest that an optimal condition to investigate the ability of 1,25-(OH)2-D3 to inhibit keratinocyte proliferation is at preconfluent cell density in the presence of KGM supplemented with 1.5 mM calcium without a feeder layer. These conditions are not appropriate for investigating the enhancement of differentiation by 1,25-(OH)2-D3, but can be used to assay other agents that modulate keratinocyte proliferation.

Versatile Genetic Tool Box for the Crenarchaeote Sulfolobus acidocaldarius

PubMed Central

Wagner, Michaela; van Wolferen, Marleen; Wagner, Alexander; Lassak, Kerstin; Meyer, Benjamin H.; Reimann, Julia; Albers, Sonja-Verena

2012-01-01

For reverse genetic approaches inactivation or selective modification of genes are required to elucidate their putative function. Sulfolobus acidocaldarius is a thermoacidophilic Crenarchaeon which grows optimally at 76°C and pH 3. As many antibiotics do not withstand these conditions the development of a genetic system in this organism is dependent on auxotrophies. Therefore we constructed a pyrE deletion mutant of S. acidocaldarius wild type strain DSM639 missing 322 bp called MW001. Using this strain as the starting point, we describe here different methods using single as well as double crossover events to obtain markerless deletion mutants, tag genes genomically and ectopically integrate foreign DNA into MW001. These methods enable us to construct single, double, and triple deletions strains that can still be complemented with the pRN1 based expression vector. Taken together we have developed a versatile and robust genetic tool box for the crenarchaeote S. acidocaldarius that will promote the study of unknown gene functions in this organism and makes it a suitable host for synthetic biology approaches. PMID:22707949

SU-G-JeP4-03: Anomaly Detection of Respiratory Motion by Use of Singular Spectrum Analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kotoku, J; Kumagai, S; Nakabayashi, S

Purpose: The implementation and realization of automatic anomaly detection of respiratory motion is a very important technique to prevent accidental damage during radiation therapy. Here, we propose an automatic anomaly detection method using singular value decomposition analysis. Methods: The anomaly detection procedure consists of four parts:1) measurement of normal respiratory motion data of a patient2) calculation of a trajectory matrix representing normal time-series feature3) real-time monitoring and calculation of a trajectory matrix of real-time data.4) calculation of an anomaly score from the similarity of the two feature matrices. Patient motion was observed by a marker-less tracking system using a depthmore » camera. Results: Two types of motion e.g. cough and sudden stop of breathing were successfully detected in our real-time application. Conclusion: Automatic anomaly detection of respiratory motion using singular spectrum analysis was successful in the cough and sudden stop of breathing. The clinical use of this algorithm will be very hopeful. This work was supported by JSPS KAKENHI Grant Number 15K08703.« less

Unsupervised markerless 3-DOF motion tracking in real time using a single low-budget camera.

PubMed

Quesada, Luis; León, Alejandro J

2012-10-01

Motion tracking is a critical task in many computer vision applications. Existing motion tracking techniques require either a great amount of knowledge on the target object or specific hardware. These requirements discourage the wide spread of commercial applications based on motion tracking. In this paper, we present a novel three degrees of freedom motion tracking system that needs no knowledge on the target object and that only requires a single low-budget camera that can be found installed in most computers and smartphones. Our system estimates, in real time, the three-dimensional position of a nonmodeled unmarked object that may be nonrigid, nonconvex, partially occluded, self-occluded, or motion blurred, given that it is opaque, evenly colored, enough contrasting with the background in each frame, and that it does not rotate. Our system is also able to determine the most relevant object to track in the screen. Our proposal does not impose additional constraints, therefore it allows a market-wide implementation of applications that require the estimation of the three position degrees of freedom of an object.

Construction and application of 3D model sequence to illustrate the development of the human embryo

NASA Astrophysics Data System (ADS)

Mizuta, Shinobu; Kakusho, Koh; Minekura, Yutaka; Minoh, Michihiko; Nakatsu, Tomoko; Shiota, Kohei

2002-05-01

Embryology is one of the basic subjects in medical education, to learn the process of human development especially from fertilization to birth. The shape deformation in the development of human embryo is one of the most important points to be comprehended, but it is difficult to illustrate the deformation by texts, 2D drawings, photographs and so on, because it is extremely complicated. The purpose of our research is to construct a 3D model sequence to illustrate the deformation of human embryo, and to make the model sequence into the teaching materials for medical education. Firstly, 3D images of the specimens of human embryo were acquired using MR microscopy. Next, an initial 3D model sequence was manually modified by comparing with the features of the acquired images under the supervision of medical doctors, because the images were influenced not only by the noise or limitation of resolution in MR image acquisition, but also by the variation of shape depending on the difference of subject. Using the constructed 3D model sequence, CG animations and an interactive VRML system were composed as the teaching materials for embryology. These materials were quite helpful to understand the shape deformation compared with the conventional materials.

Exploring personality traits related to dopamine D2/3 receptor availability in striatal subregions of humans.

PubMed

Caravaggio, Fernando; Fervaha, Gagan; Chung, Jun Ku; Gerretsen, Philip; Nakajima, Shinichiro; Plitman, Eric; Iwata, Yusuke; Wilson, Alan; Graff-Guerrero, Ariel

2016-04-01

While several studies have examined how particular personality traits are related to dopamine D2/3 receptor (D2/3R) availability in the striatum of humans, few studies have reported how multiple traits measured in the same persons are differentially related to D2/3R availability in different striatal sub-regions. We examined how personality traits measured with the Karolinska Scales of Personality are related to striatal D2/3R availability measured with [(11)C]-raclopride in 30 healthy humans. Based on previous the literature, five personality traits were hypothesized to be most likely related to D2/3R availability: impulsiveness, monotony avoidance, detachment, social desirability, and socialization. We found self-reported impulsiveness was negatively correlated with D2/3R availability in the ventral striatum and globus pallidus. After controlling for age and gender, monotony avoidance was also negatively correlated with D2/3R availability in the ventral striatum and globus pallidus. Socialization was positively correlated with D2/3R availability in the ventral striatum and putamen. After controlling for age and gender, the relationship between socialization and D2/3R availability in these regions survived correction for multiple comparisons (p-threshold=.003). Thus, within the same persons, different personality traits are differentially related to in vivo D2/3R availability in different striatal sub-regions. Copyright © 2016 Elsevier B.V. and ECNP. All rights reserved.

Dynamic three-dimensional model of the coronary circulation

NASA Astrophysics Data System (ADS)

Lehmann, Glen; Gobbi, David G.; Dick, Alexander J.; Starreveld, Yves P.; Quantz, M.; Holdsworth, David W.; Drangova, Maria

2001-05-01

A realistic numerical three-dimensional (3D) model of the dynamics of human coronary arteries has been developed. High- resolution 3D images of the coronary arteries of an excised human heart were obtained using a C-arm based computed tomography (CT) system. Cine bi-plane coronary angiograms were then acquired from a patient with similar coronary anatomy. These angiograms were used to determine the vessel motion, which was applied to the static 3D coronary tree. Corresponding arterial bifurcations were identified in the 3D CT image and in the 2D angiograms. The 3D positions of the angiographic landmarks, which were known throughout the cardiac cycle, were used to warp the 3D image via a non-linear thin-plate spline algorithm. The result was a set or 30 dynamic volumetric images sampling a complete cardiac cycle. To the best of our knowledge, the model presented here is the first dynamic 3D model that provides a true representation of both the geometry and motion of a human coronary artery tree. In the future, similar models can be generated to represent different coronary anatomy and motion. Such models are expected to become an invaluable tool during the development of dynamic imaging techniques such as MRI, multi-slice CT and 3D angiography.

Technical note: 3D from standard digital photography of human crania-a preliminary assessment.

PubMed

Katz, David; Friess, Martin

2014-05-01

This study assessed three-dimensional (3D) photogrammetry as a tool for capturing and quantifying human skull morphology. While virtual reconstruction with 3D surface scanning technology has become an accepted part of the paleoanthropologist's tool kit, recent advances in 3D photogrammetry make it a potential alternative to dedicated surface scanners. The principal advantages of photogrammetry are more rapid raw data collection, simplicity and portability of setup, and reduced equipment costs. We tested the precision and repeatability of 3D photogrammetry by comparing digital models of human crania reconstructed from conventional, 2D digital photographs to those generated using a 3D surface scanner. Overall, the photogrammetry and scanner meshes showed low degrees of deviation from one another. Surface area estimates derived from photogrammetry models tended to be slightly larger. Landmark configurations generally did not cluster together based upon whether the reconstruction was created with photogrammetry or surface scanning technology. Average deviations of landmark coordinates recorded on photogrammetry models were within the generally allowable range of error in osteometry. Thus, while dependent upon the needs of the particular research project, 3D photogrammetry appears to be a suitable, lower-cost alternative to 3D imaging and scanning options. Copyright © 2014 Wiley Periodicals, Inc.

Biofidelic Human Activity Modeling and Simulation with Large Variability

DTIC Science & Technology

2014-11-25

A systematic approach was developed for biofidelic human activity modeling and simulation by using body scan data and motion capture data to...replicate a human activity in 3D space. Since technologies for simultaneously capturing human motion and dynamic shapes are not yet ready for practical use, a...that can replicate a human activity in 3D space with the true shape and true motion of a human. Using this approach, a model library was built to

In Vivo Chondrogenesis in 3D Bioprinted Human Cell-laden Hydrogel Constructs

PubMed Central

Möller, Thomas; Hägg, Daniel; Brantsing, Camilla; Rotter, Nicole; Apelgren, Peter; Lindahl, Anders; Kölby, Lars; Gatenholm, Paul

2017-01-01

Background: The three-dimensional (3D) bioprinting technology allows creation of 3D constructs in a layer-by-layer fashion utilizing biologically relevant materials such as biopolymers and cells. The aim of this study is to investigate the use of 3D bioprinting in a clinically relevant setting to evaluate the potential of this technique for in vivo chondrogenesis. Methods: Thirty-six nude mice (Balb-C, female) received a 5- × 5- × 1-mm piece of bioprinted cell-laden nanofibrillated cellulose/alginate construct in a subcutaneous pocket. Four groups of printed constructs were used: (1) human (male) nasal chondrocytes (hNCs), (2) human (female) bone marrow–derived mesenchymal stem cells (hBMSCs), (3) coculture of hNCs and hBMSCs in a 20/80 ratio, and (4) Cell-free scaffolds (blank). After 14, 30, and 60 days, the scaffolds were harvested for histological, immunohistochemical, and mechanical analysis. Results: The constructs had good mechanical properties and keep their structural integrity after 60 days of implantation. For both the hNC constructs and the cocultured constructs, a gradual increase of glycosaminoglycan production and hNC proliferation was observed. However, the cocultured group showed a more pronounced cell proliferation and enhanced deposition of human collagen II demonstrated by immunohistochemical analysis. Conclusions: In vivo chondrogenesis in a 3D bioprinted human cell-laden hydrogel construct has been demonstrated. The trophic role of the hBMSCs in stimulating hNC proliferation and matrix deposition in the coculture group suggests the potential of 3D bioprinting of human cartilage for future application in reconstructive surgery. PMID:28280669

In Vivo Chondrogenesis in 3D Bioprinted Human Cell-laden Hydrogel Constructs.

PubMed

Möller, Thomas; Amoroso, Matteo; Hägg, Daniel; Brantsing, Camilla; Rotter, Nicole; Apelgren, Peter; Lindahl, Anders; Kölby, Lars; Gatenholm, Paul

2017-02-01

The three-dimensional (3D) bioprinting technology allows creation of 3D constructs in a layer-by-layer fashion utilizing biologically relevant materials such as biopolymers and cells. The aim of this study is to investigate the use of 3D bioprinting in a clinically relevant setting to evaluate the potential of this technique for in vivo chondrogenesis. Thirty-six nude mice (Balb-C, female) received a 5- × 5- × 1-mm piece of bioprinted cell-laden nanofibrillated cellulose/alginate construct in a subcutaneous pocket. Four groups of printed constructs were used: (1) human (male) nasal chondrocytes (hNCs), (2) human (female) bone marrow-derived mesenchymal stem cells (hBMSCs), (3) coculture of hNCs and hBMSCs in a 20/80 ratio, and (4) Cell-free scaffolds (blank). After 14, 30, and 60 days, the scaffolds were harvested for histological, immunohistochemical, and mechanical analysis. The constructs had good mechanical properties and keep their structural integrity after 60 days of implantation. For both the hNC constructs and the cocultured constructs, a gradual increase of glycosaminoglycan production and hNC proliferation was observed. However, the cocultured group showed a more pronounced cell proliferation and enhanced deposition of human collagen II demonstrated by immunohistochemical analysis. In vivo chondrogenesis in a 3D bioprinted human cell-laden hydrogel construct has been demonstrated. The trophic role of the hBMSCs in stimulating hNC proliferation and matrix deposition in the coculture group suggests the potential of 3D bioprinting of human cartilage for future application in reconstructive surgery.

Rotigotine is a potent agonist at dopamine D1 receptors as well as at dopamine D2 and D3 receptors.

PubMed

Wood, Martyn; Dubois, Vanessa; Scheller, Dieter; Gillard, Michel

2015-02-01

Rotigotine acts as a dopamine receptor agonist with high affinity for the dopamine D2, D3, D4 and D5 receptors but with a low affinity for the dopamine D1 receptor. We have investigated this further in radioligand binding and functional studies and compared the profile of rotigotine with that of other drugs used in the treatment of Parkinson's disease (PD). The binding of rotigotine to human dopamine D1, D2, D3, D4 and D5 receptors was determined in radioligand binding studies using [(3)H]rotigotine and compared with that of standard antagonist radioligands. Functional interactions of rotigotine with human dopamine receptors was also determined. [(3)H]rotigotine can be used as an agonist radioligand to label all dopamine receptor subtypes and this can be important to derive agonist affinity estimates. Rotigotine maintains this high affinity in functional studies at all dopamine receptors especially D1, D2 and D3 receptors and, to a lesser extent, D4 and D5 receptors. Rotigotine, like apomorphine but unlike ropinirole and pramipexole, was a potent agonist at all dopamine receptors. Rotigotine is a high-potency agonist at human dopamine D1, D2 and D3 receptors with a lower potency at D4 and D5 receptors. These studies differentiate rotigotine from conventional dopamine D2 agonists, used in the treatment of PD, such as ropinirole and pramipexole which lack activity at the D1 and D5 receptors, but resembles that of apomorphine which has greater efficacy in PD than other dopamine agonists but has suboptimal pharmacokinetic properties. © 2014 The British Pharmacological Society.

Rotigotine is a potent agonist at dopamine D1 receptors as well as at dopamine D2 and D3 receptors

PubMed Central

Wood, Martyn; Dubois, Vanessa; Scheller, Dieter; Gillard, Michel

2015-01-01

Background and Purpose Rotigotine acts as a dopamine receptor agonist with high affinity for the dopamine D2, D3, D4 and D5 receptors but with a low affinity for the dopamine D1 receptor. We have investigated this further in radioligand binding and functional studies and compared the profile of rotigotine with that of other drugs used in the treatment of Parkinson's disease (PD). Experimental Approach The binding of rotigotine to human dopamine D1, D2, D3, D4 and D5 receptors was determined in radioligand binding studies using [3H]rotigotine and compared with that of standard antagonist radioligands. Functional interactions of rotigotine with human dopamine receptors was also determined. Key Results [3H]rotigotine can be used as an agonist radioligand to label all dopamine receptor subtypes and this can be important to derive agonist affinity estimates. Rotigotine maintains this high affinity in functional studies at all dopamine receptors especially D1, D2 and D3 receptors and, to a lesser extent, D4 and D5 receptors. Rotigotine, like apomorphine but unlike ropinirole and pramipexole, was a potent agonist at all dopamine receptors. Conclusions and Implications Rotigotine is a high-potency agonist at human dopamine D1, D2 and D3 receptors with a lower potency at D4 and D5 receptors. These studies differentiate rotigotine from conventional dopamine D2 agonists, used in the treatment of PD, such as ropinirole and pramipexole which lack activity at the D1 and D5 receptors, but resembles that of apomorphine which has greater efficacy in PD than other dopamine agonists but has suboptimal pharmacokinetic properties. PMID:25339241

Megalin-Mediated Endocytosis of Vitamin D Binding Protein Correlates with 25-Hydroxycholecalciferol Actions in Human Mammary Cells1

PubMed Central

Rowling, Matthew J.; Kemmis, Carly M.; Taffany, David A.; Welsh, JoEllen

2007-01-01

The major circulating form of vitamin D is 25-hydroxycholecalciferol [25(OH)D3], which is delivered to target tissues in complex with the serum vitamin D binding protein (DBP). We recently observed that mammary cells can metabolize 25(OH)D3 to 1,25-dihydroxycholecalciferol [1,25(OH)2D3], the vitamin D receptor (VDR) ligand, and the objective of our study was to elucidate the mechanisms by which the 25(OH)D3-DBP complex is internalized by mammary cells prior to metabolism. Using fluorescent microscopy and temperature-shift techniques, we found that T-47D breast cancer cells rapidly internalize DBP via endocytosis, which is blunted by receptor-associated protein, a specific inhibitor of megalin-mediated endocytosis. Endocytosis of DBP was associated with activation of VDR by 25(OH)D3 but not 1,25(OH)2D3 (as measured by induction of the VDR target gene, CYP24). We also found that megalin and its endocytic partner, cubilin, are coexpressed in normal murine mammary tissue, in nontransformed human mammary epithelial cell lines, and in some established human breast cancer cell lines. To our knowledge, our studies are the first to demonstrate that mammary-derived cells express megalin and cubilin, which contribute to the endocytic uptake of 25(OH)D3-DBP and activation of the VDR pathway. PMID:17056796

Multifunctional Bioreactor System for Human Intestine Tissues

PubMed Central

2017-01-01

The three-dimensional (3D) cultivation of intestinal cells and tissues in dynamic bioreactor systems to represent in vivo intestinal microenvironments is essential for developing regenerative medicine treatments for intestinal diseases. We have previously developed in vitro human intestinal tissue systems using a 3D porous silk scaffold system with intestinal architectures and topographical features for the adhesion, growth, and differentiation of intestinal cells under static culture conditions. In this study, we designed and fabricated a multifunctional bioreactor system that incorporates pre-epithelialized 3D silk scaffolds in a dynamic culture environment for in vitro engineering of human intestine tissues. The bioreactor system allows for control of oxygen levels in perfusion fluids (aerobic simulated intestinal fluid (SIF), microaerobic SIF, and anaerobic SIF), while ensuring control over the mechanical and chemical microenvironments present in native human intestines. The bioreactor system also enables 3D cell culture with spatial separation and cultivation of cocultured epithelial and stromal cells. Preliminary functional analysis of tissues housed in the bioreactor demonstrated that the 3D tissue constructs survived and maintained typical phenotypes of intestinal epithelium, including epithelial tight junction formation, intestinal biomarker expression, microvilli formation, and mucus secretion. The unique combination of a dynamic bioreactor and 3D intestinal constructs offers utility for engineering human intestinal tissues for the study of intestinal diseases and discovery options for new treatments. PMID:29333491

[Establishment of a 3D finite element model of human skull using MSCT images and mimics software].

PubMed

Huang, Ping; Li, Zheng-dong; Shao, Yu; Zou, Dong-hua; Liu, Ning-guo; Li, Li; Chen, Yuan-yuan; Wan, Lei; Chen, Yi-jiu

2011-02-01

To establish a human 3D finite element skull model, and to explore its value in biomechanics analysis. The cadaveric head was scanned and then 3D skull model was created using Mimics software based on 2D CT axial images. The 3D skull model was optimized by preprocessor along with creation of the surface and volume meshes. The stress changes, after the head was struck by an object or the head hit the ground directly, were analyzed using ANSYS software. The original 3D skull model showed a large number of triangles with a poor quality and high similarity with the real head, while the optimized model showed high quality surface and volume meshes with a small number of triangles comparatively. The model could show the local and global stress changes effectively. The human 3D skull model can be established using MSCT and Mimics software and provides a good finite element model for biomechanics analysis. This model may also provide a base for the study of head stress changes following different forces.

3D Digitization and Prototyping of the Skull for Practical Use in the Teaching of Human Anatomy.

PubMed

Lozano, Maria Teresa Ugidos; Haro, Fernando Blaya; Diaz, Carlos Molino; Manzoor, Sadia; Ugidos, Gonzalo Ferrer; Mendez, Juan Antonio Juanes

2017-05-01

The creation of new rapid prototyping techniques, low cost 3D printers as well as the creation of new software for these techniques have allowed the creation of 3D models of bones making their application possible in the field of teaching anatomy in the faculties of Health Sciences. The 3D model of cranium created in the present work, at full scale, present accurate reliefs and anatomical details that are easily identifiable by undergraduate students in their use for the study of human anatomy. In this article, the process of scanning the skull and the subsequent treatment of these images with specific software until the generation of 3D model using 3D printer has been reported.

Serotype-specific differences in inhibition of reovirus infectivity by human-milk glycans are determined by viral attachment protein σ1.

PubMed

Iskarpatyoti, Jason A; Morse, E Ashley; McClung, R Paul; Ikizler, Miné; Wetzel, J Denise; Contractor, Nikhat; Dermody, Terence S

2012-11-25

Human milk contains many bioactive components, including secretory IgA, oligosaccharides, and milk-associated proteins. We assessed the antiviral effects of several components of milk against mammalian reoviruses. We found that glucocerebroside (GCB) inhibited the infectivity of reovirus strain type 1 Lang (T1L), whereas gangliosides GD3 and GM3 and 3'-sialyllactose (3SL) inhibited the infectivity of reovirus strain type 3 Dearing (T3D). Agglutination of erythrocytes mediated by T1L and T3D was inhibited by GD3, GM3, and bovine lactoferrin. Additionally, α-sialic acid, 3SL, 6'-sialyllactose, sialic acid, human lactoferrin, osteopontin, and α-lactalbumin inhibited hemagglutination mediated by T3D. Using single-gene reassortant viruses, we found that serotype-specific differences segregate with the gene encoding the viral attachment protein. Furthermore, GD3, GM3, and 3SL inhibit T3D infectivity by blocking binding to host cells, whereas GCB inhibits T1L infectivity post-attachment. These results enhance an understanding of reovirus cell attachment and define a mechanism for the antimicrobial activity of human milk. Copyright © 2012 Elsevier Inc. All rights reserved.

Caffeine increases striatal dopamine D2/D3 receptor availability in the human brain.

PubMed

Volkow, N D; Wang, G-J; Logan, J; Alexoff, D; Fowler, J S; Thanos, P K; Wong, C; Casado, V; Ferre, S; Tomasi, D

2015-04-14

Caffeine, the most widely consumed psychoactive substance in the world, is used to promote wakefulness and enhance alertness. Like other wake-promoting drugs (stimulants and modafinil), caffeine enhances dopamine (DA) signaling in the brain, which it does predominantly by antagonizing adenosine A2A receptors (A2AR). However, it is unclear if caffeine, at the doses consumed by humans, increases DA release or whether it modulates the functions of postsynaptic DA receptors through its interaction with adenosine receptors, which modulate them. We used positron emission tomography and [(11)C]raclopride (DA D2/D3 receptor radioligand sensitive to endogenous DA) to assess if caffeine increased DA release in striatum in 20 healthy controls. Caffeine (300 mg p.o.) significantly increased the availability of D2/D3 receptors in putamen and ventral striatum, but not in caudate, when compared with placebo. In addition, caffeine-induced increases in D2/D3 receptor availability in the ventral striatum were associated with caffeine-induced increases in alertness. Our findings indicate that in the human brain, caffeine, at doses typically consumed, increases the availability of DA D2/D3 receptors, which indicates that caffeine does not increase DA in the striatum for this would have decreased D2/D3 receptor availability. Instead, we interpret our findings to reflect an increase in D2/D3 receptor levels in striatum with caffeine (or changes in affinity). The association between increases in D2/D3 receptor availability in ventral striatum and alertness suggests that caffeine might enhance arousal, in part, by upregulating D2/D3 receptors.

Caffeine increases striatal dopamine D2/D3 receptor availability in the human brain

PubMed Central

Volkow, N D; Wang, G-J; Logan, J; Alexoff, D; Fowler, J S; Thanos, P K; Wong, C; Casado, V; Ferre, S; Tomasi, D

2015-01-01

Caffeine, the most widely consumed psychoactive substance in the world, is used to promote wakefulness and enhance alertness. Like other wake-promoting drugs (stimulants and modafinil), caffeine enhances dopamine (DA) signaling in the brain, which it does predominantly by antagonizing adenosine A2A receptors (A2AR). However, it is unclear if caffeine, at the doses consumed by humans, increases DA release or whether it modulates the functions of postsynaptic DA receptors through its interaction with adenosine receptors, which modulate them. We used positron emission tomography and [11C]raclopride (DA D2/D3 receptor radioligand sensitive to endogenous DA) to assess if caffeine increased DA release in striatum in 20 healthy controls. Caffeine (300 mg p.o.) significantly increased the availability of D2/D3 receptors in putamen and ventral striatum, but not in caudate, when compared with placebo. In addition, caffeine-induced increases in D2/D3 receptor availability in the ventral striatum were associated with caffeine-induced increases in alertness. Our findings indicate that in the human brain, caffeine, at doses typically consumed, increases the availability of DA D2/D3 receptors, which indicates that caffeine does not increase DA in the striatum for this would have decreased D2/D3 receptor availability. Instead, we interpret our findings to reflect an increase in D2/D3 receptor levels in striatum with caffeine (or changes in affinity). The association between increases in D2/D3 receptor availability in ventral striatum and alertness suggests that caffeine might enhance arousal, in part, by upregulating D2/D3 receptors. PMID:25871974

Caffeine increases striatal dopamine D 2/D 3 receptor availability in the human brain

DOE PAGES

Volkow, N. D.; Wang, G. -J.; Logan, J.; ...

2015-04-14

Caffeine, the most widely consumed psychoactive substance in the world, is used to promote wakefulness and enhance alertness. Like other wake-promoting drugs (stimulants and modafinil), caffeine enhances dopamine (DA) signaling in the brain, which it does predominantly by antagonizing adenosine A 2A receptors (A 2AR). However, it is unclear if caffeine, at the doses consumed by humans, increases DA release or whether it modulates the functions of postsynaptic DA receptors through its interaction with adenosine receptors, which modulate them. We used positron emission tomography and [ 11C]raclopride (DA D 2/D 3 receptor radioligand sensitive to endogenous DA) to assess ifmore » caffeine increased DA release in striatum in 20 healthy controls. Caffeine (300mg p.o.) significantly increased the availability of D 2/D 3 receptors in putamen and ventral striatum, but not in caudate, when compared with placebo. In addition, caffeine-induced increases in D 2/D 3 receptor availability in the ventral striatum were associated with caffeine-induced increases in alertness. Our findings indicate that in the human brain, caffeine, at doses typically consumed, increases the availability of DA D 2/D 3 receptors, which indicates that caffeine does not increase DA in the striatum for this would have decreased D 2/D 3 receptor availability. Instead, we interpret our findings to reflect an increase in D 2/D 3 receptor levels in striatum with caffeine (or changes in affinity). Furthermore, the association between increases in D 2/D 3 receptor availability in ventral striatum and alertness suggests that caffeine might enhance arousal, in part, by upregulating D 2/D 3 receptors.« less

Caffeine increases striatal dopamine D 2/D 3 receptor availability in the human brain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Volkow, N. D.; Wang, G. -J.; Logan, J.

Caffeine, the most widely consumed psychoactive substance in the world, is used to promote wakefulness and enhance alertness. Like other wake-promoting drugs (stimulants and modafinil), caffeine enhances dopamine (DA) signaling in the brain, which it does predominantly by antagonizing adenosine A 2A receptors (A 2AR). However, it is unclear if caffeine, at the doses consumed by humans, increases DA release or whether it modulates the functions of postsynaptic DA receptors through its interaction with adenosine receptors, which modulate them. We used positron emission tomography and [ 11C]raclopride (DA D 2/D 3 receptor radioligand sensitive to endogenous DA) to assess ifmore » caffeine increased DA release in striatum in 20 healthy controls. Caffeine (300mg p.o.) significantly increased the availability of D 2/D 3 receptors in putamen and ventral striatum, but not in caudate, when compared with placebo. In addition, caffeine-induced increases in D 2/D 3 receptor availability in the ventral striatum were associated with caffeine-induced increases in alertness. Our findings indicate that in the human brain, caffeine, at doses typically consumed, increases the availability of DA D 2/D 3 receptors, which indicates that caffeine does not increase DA in the striatum for this would have decreased D 2/D 3 receptor availability. Instead, we interpret our findings to reflect an increase in D 2/D 3 receptor levels in striatum with caffeine (or changes in affinity). Furthermore, the association between increases in D 2/D 3 receptor availability in ventral striatum and alertness suggests that caffeine might enhance arousal, in part, by upregulating D 2/D 3 receptors.« less

A New Human 3D-Liver Model Unravels the Role of Galectins in Liver Infection by the Parasite Entamoeba histolytica

PubMed Central

Petropolis, Debora B.; Faust, Daniela M.; Deep Jhingan, Gagan; Guillen, Nancy

2014-01-01

Investigations of human parasitic diseases depend on the availability of appropriate in vivo animal models and ex vivo experimental systems, and are particularly difficult for pathogens whose exclusive natural hosts are humans, such as Entamoeba histolytica, the protozoan parasite responsible for amoebiasis. This common infectious human disease affects the intestine and liver. In the liver sinusoids E. histolytica crosses the endothelium and penetrates into the parenchyma, with the concomitant initiation of inflammatory foci and subsequent abscess formation. Studying factors responsible for human liver infection is hampered by the complexity of the hepatic environment and by the restrictions inherent to the use of human samples. Therefore, we built a human 3D-liver in vitro model composed of cultured liver sinusoidal endothelial cells and hepatocytes in a 3D collagen-I matrix sandwich. We determined the presence of important hepatic markers and demonstrated that the cell layers function as a biological barrier. E. histolytica invasion was assessed using wild-type strains and amoebae with altered virulence or different adhesive properties. We showed for the first time the dependence of endothelium crossing upon amoebic Gal/GalNAc lectin. The 3D-liver model enabled the molecular analysis of human cell responses, suggesting for the first time a crucial role of human galectins in parasite adhesion to the endothelial cells, which was confirmed by siRNA knockdown of galectin-1. Levels of several pro-inflammatory cytokines, including galectin-1 and -3, were highly increased upon contact of E. histolytica with the 3D-liver model. The presence of galectin-1 and -3 in the extracellular medium stimulated pro-inflammatory cytokine release, suggesting a further role for human galectins in the onset of the hepatic inflammatory response. These new findings are relevant for a better understanding of human liver infection by E. histolytica. PMID:25211477

DEVELOPMENT OF 3-D COMPUTER MODELS OF HUMAN LUNG MORPHOLOGY FOR IMPROOVED RISK ASSESSMENT OF INHALED PARTICULATE MATTER

EPA Science Inventory

DEVELOPMENT OF 3-D COMPUTER MODELS OF HUMAN LUNG MORPHOLOGY FOR IMPROVED RISK ASSESSMENT OF INHALED PARTICULATE MATTER

Jeffry D. Schroeter, Curriculum in Toxicology, University of North Carolina, Chapel Hill, NC 27599; Ted B. Martonen, ETD, NHEERL, USEPA, RTP, NC 27711; Do...

Development of 3D imaging technique of reconstructed human epidermis with immortalized human epidermal cell line.

PubMed

Inoue, Yu; Hasegawa, Seiji; Miyachi, Katsuma; Yamada, Takaaki; Nakata, Satoru; Ipponjima, Sari; Hibi, Terumasa; Nemoto, Tomomi; Tanaka, Masahiko; Suzuki, Ryo; Hirashima, Naohide

2018-05-01

The epidermis, the outermost layer of the skin, retains moisture and functions as a physical barrier against the external environment. Epidermal cells are continuously replaced by turnover, and thus to understand in detail the dynamic cellular events in the epidermis, techniques to observe live tissues in 3D are required. Here, we established a live 3D imaging technique for epidermis models. We first obtained immortalized human epidermal cell lines which have a normal differentiation capacity and fluorescence-labelled cytoplasm or nuclei. The reconstituted 3D epidermis was prepared with these lines. Using this culture system, we were able to observe the structure of the reconstituted epidermis live in 3D, which was similar to an in vivo epidermis, and evaluate the effect of a skin irritant. This technique may be useful for dermatological science and drug development. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

[3D evaluation model for drug hepatotoxicity testing on HepG2 cells and its application in drug safety evaluation].

PubMed

Li, Dan-Dan; Tang, Xiang-Lin; Tan, Hong-Ling; Liang, Qian-de; Wang, Yu-Guang; Ma, Zeng-Chun; Xiao, Cheng-Rong; Gao, Yue

2016-04-01

3D in vitro toxicity testing model was developed by magnetic levitation method for culture of the human hepatoma cell line HepG2 and applied to evaluate the drug hepatotoxicity. After formation of stable 3D structure for HepG2 cells, their glycogen storage capacity under 2D and 3D culture conditions were detected by immunohistochemistry technology, and the mRNA expression levels of phase Ⅰ and Ⅱ drug metabolism enzymes, drug transporters, nuclear receptors and liver-specific marker albumin(ALB) were compared between 2D and 3D culture conditions by using RT-PCR method. Immunohistochemistry results showed that HepG2 cells had abundant glycogen storage capacity under 3D culture conditions, which was similar to human liver tissues. The mRNA expression levels of major drug metabolism enzymes, drug transporters, nuclear receptors and ALB in HepG2 cells under 3D culture conditions were up-regulated as compared with 2D culture conditions. For drug hepatotoxicity evaluation, the typical hepatotoxic drug acetaminophen(APAP), and most reported drugs Polygonum multiflorum Thunb.(Chinese name He-shou-wu) and Psoraleae corylifolia L.(Chinese name Bu-gu-zhi) were selected for single dose and repeated dose(7 d) exposure. In the repeated dose exposure test, 3D HepG2 cells showed higher sensitivity. This established 3D HepG2 cells model with magnetic levitation 3D culture techniques was more close to the human liver tissues both in morphology and functions, so it was a better 3D hepatotoxicity evaluation model. Copyright© by the Chinese Pharmaceutical Association.

Bioengineering Anabolic Vitamin D-25-Hydroxylase Activity into the Human Vitamin D Catabolic Enzyme, Cytochrome P450 CYP24A1, by a V391L Mutation*

PubMed Central

Kaufmann, Martin; Prosser, David E.; Jones, Glenville

2011-01-01

CYP24A1 is a mitochondrial cytochrome P450 (CYP) that catabolizes 1α,25-dihydroxyvitamin D3 (1α,25-(OH)2D3) to different products: calcitroic acid or 1α,25-(OH)2D3-26,23-lactone via multistep pathways commencing with C24 and C23 hydroxylation, respectively. Despite the ability of CYP24A1 to catabolize a wide range of 25-hydroxylated analogs including 25-hydroxyvitamin D3, the enzyme is unable to metabolize the synthetic prodrug, 1α-hydroxyvitamin D3 (1α-OH-D3), presumably because it lacks a C25-hydroxyl. In the current study we show that a single V391L amino acid substitution in the β3a-strand of human CYP24A1 converts this enzyme from a catabolic 1α,25-(OH)2D3-24-hydroxylase into an anabolic 1α-OH-D3-25-hydroxylase, thereby forming the hormone, 1α,25-(OH)2D3. Furthermore, because the mutant enzyme retains its basal ability to catabolize 1α,25-(OH)2D3 via C24 hydroxylation, it can also make calcitroic acid. Previous work has shown that an A326G mutation is responsible for the regioselectivity differences observed between human (primarily C24-hydroxylating) and opossum (C23-hydroxylating) CYP24A1. When the V391L and A326G mutations were combined (V391L/A326G), the mutant enzyme continued to form 1α,25-(OH)2D3 from 1α-OH-D3, but this initial product was diverted via the C23 hydroxylation pathway into the 26,23-lactone. The relative position of Val-391 in the β3a-strand of a homology model and the crystal structure of rat CYP24A1 is consistent with hydrophobic contact of Val-391 and the substrate side chain near C21. We interpret that the substrate specificity of V391L-modified human CYP24A1 toward 1α-OH-D3 is enabled by an altered contact with the substrate side chain that optimally positions C25 of the 1α-OH-D3 above the heme for hydroxylation. PMID:21697097

The 3D Human Motion Control Through Refined Video Gesture Annotation

NASA Astrophysics Data System (ADS)

Jin, Yohan; Suk, Myunghoon; Prabhakaran, B.

In the beginning of computer and video game industry, simple game controllers consisting of buttons and joysticks were employed, but recently game consoles are replacing joystick buttons with novel interfaces such as the remote controllers with motion sensing technology on the Nintendo Wii [1] Especially video-based human computer interaction (HCI) technique has been applied to games, and the representative game is 'Eyetoy' on the Sony PlayStation 2. Video-based HCI technique has great benefit to release players from the intractable game controller. Moreover, in order to communicate between humans and computers, video-based HCI is very crucial since it is intuitive, easy to get, and inexpensive. On the one hand, extracting semantic low-level features from video human motion data is still a major challenge. The level of accuracy is really dependent on each subject's characteristic and environmental noises. Of late, people have been using 3D motion-capture data for visualizing real human motions in 3D space (e.g, 'Tiger Woods' in EA Sports, 'Angelina Jolie' in Bear-Wolf movie) and analyzing motions for specific performance (e.g, 'golf swing' and 'walking'). 3D motion-capture system ('VICON') generates a matrix for each motion clip. Here, a column is corresponding to a human's sub-body part and row represents time frames of data capture. Thus, we can extract sub-body part's motion only by selecting specific columns. Different from low-level feature values of video human motion, 3D human motion-capture data matrix are not pixel values, but is closer to human level of semantics.

Mouse and human BAC transgenes recapitulate tissue-specific expression of the vitamin D receptor in mice and rescue the VDR-null phenotype.

PubMed

Lee, Seong Min; Bishop, Kathleen A; Goellner, Joseph J; O'Brien, Charles A; Pike, J Wesley

2014-06-01

The biological actions of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) are mediated by the vitamin D receptor (VDR), which is expressed in numerous target tissues in a cell type-selective manner. Recent studies using genomic analyses and recombineered bacterial artificial chromosomes (BACs) have defined the specific features of mouse and human VDR gene loci in vitro. In the current study, we introduced recombineered mouse and human VDR BACs as transgenes into mice and explored their expression capabilities in vivo. Individual transgenic mouse strains selectively expressed BAC-derived mouse or human VDR proteins in appropriate vitamin D target tissues, thereby recapitulating the tissue-specific expression of endogenous mouse VDR. The mouse VDR transgene was also regulated by 1,25(OH)2D3 and dibutyryl-cAMP. When crossed into a VDR-null mouse background, both transgenes restored wild-type basal as well as 1,25(OH)2D3-inducible gene expression patterns in the appropriate tissues. This maneuver resulted in the complete rescue of the aberrant phenotype noted in the VDR-null mouse, including systemic features associated with altered calcium and phosphorus homeostasis and disrupted production of parathyroid hormone and fibroblast growth factor 23, and abnormalities associated with the skeleton, kidney, parathyroid gland, and the skin. This study suggests that both mouse and human VDR transgenes are capable of recapitulating basal and regulated expression of the VDR in the appropriate mouse tissues and restore 1,25(OH)2D3 function. These results provide a baseline for further dissection of mechanisms integral to mouse and human VDR gene expression and offer the potential to explore the consequence of selective mutations in VDR proteins in vivo.

Recognizing an individual face: 3D shape contributes earlier than 2D surface reflectance information.

PubMed

Caharel, Stéphanie; Jiang, Fang; Blanz, Volker; Rossion, Bruno

2009-10-01

The human brain recognizes faces by means of two main diagnostic sources of information: three-dimensional (3D) shape and two-dimensional (2D) surface reflectance. Here we used event-related potentials (ERPs) in a face adaptation paradigm to examine the time-course of processing for these two types of information. With a 3D morphable model, we generated pairs of faces that were either identical, varied in 3D shape only, in 2D surface reflectance only, or in both. Sixteen human observers discriminated individual faces in these 4 types of pairs, in which a first (adapting) face was followed shortly by a second (test) face. Behaviorally, observers were as accurate and as fast for discriminating individual faces based on either 3D shape or 2D surface reflectance alone, but were faster when both sources of information were present. As early as the face-sensitive N170 component (approximately 160 ms following the test face), there was larger amplitude for changes in 3D shape relative to the repetition of the same face, especially over the right occipito-temporal electrodes. However, changes in 2D reflectance between the adapter and target face did not increase the N170 amplitude. At about 250 ms, both 3D shape and 2D reflectance contributed equally, and the largest difference in amplitude compared to the repetition of the same face was found when both 3D shape and 2D reflectance were combined, in line with observers' behavior. These observations indicate that evidence to recognize individual faces accumulate faster in the right hemisphere human visual cortex from diagnostic 3D shape information than from 2D surface reflectance information.

Type 1 diabetes vaccine candidates promote human Foxp3+Treg induction in humanized mice

PubMed Central

Serr, Isabelle; Fürst, Rainer W.; Achenbach, Peter; Scherm, Martin G.; Gökmen, Füsun; Haupt, Florian; Sedlmeier, Eva-Maria; Knopff, Annette; Shultz, Leonard; Willis, Richard A.; Ziegler, Anette-Gabriele; Daniel, Carolin

2016-01-01

Immune tolerance is executed partly by Foxp3+regulatory T (Treg) cells, which suppress autoreactive T cells. In autoimmune type 1 diabetes (T1D) impaired tolerance promotes destruction of insulin-producing β-cells. The development of autoantigen-specific vaccination strategies for Foxp3+Treg-induction and prevention of islet autoimmunity in patients is still in its infancy. Here, using human haematopoietic stem cell-engrafted NSG-HLA-DQ8 transgenic mice, we provide direct evidence for human autoantigen-specific Foxp3+Treg-induction in vivo. We identify HLA-DQ8-restricted insulin-specific CD4+T cells and demonstrate efficient human insulin-specific Foxp3+Treg-induction upon subimmunogenic vaccination with strong agonistic insulin mimetopes in vivo. Induced human Tregs are stable, show increased expression of Treg signature genes such as Foxp3, CTLA4, IL-2Rα and TIGIT and can efficiently suppress effector T cells. Such Foxp3+Treg-induction does not trigger any effector T cells. These T1D vaccine candidates could therefore represent an expedient improvement in the challenge to induce human Foxp3+Tregs and to develop novel precision medicines for prevention of islet autoimmunity in children at risk of T1D. PMID:26975663

Targeting the vitamin D endocrine system (VDES) for the management of inflammatory and malignant skin diseases: An historical view and outlook.

PubMed

Reichrath, Jörg; Zouboulis, Christos C; Vogt, Thomas; Holick, Michael F

2016-09-01

Vitamin D represents one of the major driving factors for the development of life on earth and for human evolution. While up to 10-20 % of the human organism's requirements in vitamin D can be obtained by the diet (under most living conditions in the USA and Europe), approximately 90 % of all needed vitamin D has to be photosynthesized in the skin through the action of the sun (ultraviolet-B (UV-B)). The skin represents a key organ of the human body's vitamin D endocrine system (VDES), being both the site of vitamin D synthesis and a target tissue for biologically active vitamin D metabolites. It was shown that human keratinocytes possess the enzymatic machinery (CYP27B1) for the synthesis of the biologically most active natural vitamin D metabolite 1,25-dihydroxyvitamin D 3 (1,25(OH) 2 D 3 ), representing an autonomous vitamin D 3 pathway. Cutaneous production of 1,25(OH) 2 D 3 may exert intracrine, autocrine, and paracrine effects on keratinocytes and on neighboring cells. Many skin cells (including keratinocytes, sebocytes, fibroblasts, melanocytes, and skin immune cells) express the vitamin D receptor (VDR), an absolute pre-requisite for the mediation of genomic effects of 1,25(OH) 2 D 3 and analogs. VDR belongs to the superfamily of trans-acting transcriptional regulatory factors, which includes the steroid and thyroid hormone receptors as well as the retinoid X receptors (RXR) and retinoic acid receptors (RAR). Numerous studies, including cDNA microarray analyses of messenger RNAs (mRNAs), indicate that as many as 500-1000 genes may be regulated by VDR ligands that control various cellular functions including growth, differentiation, and apoptosis. The observation that 1,25(OH) 2 D 3 is extremely effective in inducing the terminal differentiation and in inhibiting the proliferation of cultured human keratinocytes has resulted in the use of vitamin D analogs for the treatment of psoriasis. This review gives an historical view and summarizes our present knowledge about the relevance of the VDES for the management of inflammatory and malignant skin diseases.

Effectiveness of three-dimensional digital animation in teaching human anatomy in an authentic classroom context.

PubMed

Hoyek, Nady; Collet, Christian; Di Rienzo, Franck; De Almeida, Mickael; Guillot, Aymeric

2014-01-01

Three-dimensional (3D) digital animations were used to teach the human musculoskeletal system to first year kinesiology students. The purpose of this study was to assess the effectiveness of this method by comparing two groups from two different academic years during two of their official required anatomy examinations (trunk and upper limb assessments). During the upper limb section, the teacher used two-dimensional (2D) drawings embedded into PowerPoint(®) slides and 3D digital animations for the first group (2D group) and the second (3D group), respectively. The same 3D digital animations were used for both groups during the trunk section. The only difference between the two was the multimedia used to present the information during the upper limb section. The 2D group surprisingly outperformed the 3D group on the trunk assessment. On the upper limb assessment no difference in the scores on the overall anatomy examination was found. However, the 3D group outperformed the 2D group in questions requiring spatial ability. Data supported that 3D digital animations were effective instructional multimedia material tools in teaching human anatomy especially in recalling anatomical knowledge requiring spatial ability. The importance of evaluating the effectiveness of a new instructional material outside laboratory environment (e.g., after a complete semester and on official examinations) was discussed. © 2014 American Association of Anatomists.

A 3D Human Renal Cell Carcinoma-on-a-Chip for the Study of Tumor Angiogenesis.

PubMed

Miller, Chris P; Tsuchida, Connor; Zheng, Ying; Himmelfarb, Jonathan; Akilesh, Shreeram

2018-06-01

Tractable human tissue-engineered 3D models of cancer that enable fine control of tumor growth, metabolism, and reciprocal interactions between different cell types in the tumor microenvironment promise to accelerate cancer research and pharmacologic testing. Progress to date mostly reflects the use of immortalized cancer cell lines, and progression to primary patient-derived tumor cells is needed to realize the full potential of these platforms. For the first time, we report endothelial sprouting induced by primary patient tumor cells in a 3D microfluidic system. Specifically, we have combined primary human clear cell renal cell carcinoma (ccRCC) cells from six independent donors with human endothelial cells in a vascularized, flow-directed, 3D culture system ("ccRCC-on-a-chip"). The upregulation of key angiogenic factors in primary human ccRCC cells, which exhibited unique patterns of donor variation, was further enhanced when they were cultured in 3D clusters. When embedded in the matrix surrounding engineered human vessels, these ccRCC tumor clusters drove potent endothelial cell sprouting under continuous flow, thus recapitulating the critical angiogenic signaling axis between human ccRCC cells and endothelial cells. Importantly, this phenotype was driven by a primary tumor cell-derived biochemical gradient of angiogenic growth factor accumulation that was subject to pharmacological blockade. Our novel 3D system represents a vascularized tumor model that is easy to image and quantify and is fully tunable in terms of input cells, perfusate, and matrices. We envision that this ccRCC-on-a-chip will be valuable for mechanistic studies, for studying tumor-vascular cell interactions, and for developing novel and personalized antitumor therapies. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Print Me an Organ? Ethical and Regulatory Issues Emerging from 3D Bioprinting in Medicine.

PubMed

Gilbert, Frederic; O'Connell, Cathal D; Mladenovska, Tajanka; Dodds, Susan

2018-02-01

Recent developments of three-dimensional printing of biomaterials (3D bioprinting) in medicine have been portrayed as demonstrating the potential to transform some medical treatments, including providing new responses to organ damage or organ failure. However, beyond the hype and before 3D bioprinted organs are ready to be transplanted into humans, several important ethical concerns and regulatory questions need to be addressed. This article starts by raising general ethical concerns associated with the use of bioprinting in medicine, then it focuses on more particular ethical issues related to experimental testing on humans, and the lack of current international regulatory directives to guide these experiments. Accordingly, this article (1) considers whether there is a limit as to what should be bioprinted in medicine; (2) examines key risks of significant harm associated with testing 3D bioprinting for humans; (3) investigates the clinical trial paradigm used to test 3D bioprinting; (4) analyses ethical questions of irreversibility, loss of treatment opportunity and replicability; (5) explores the current lack of a specific framework for the regulation and testing of 3D bioprinting treatments.

Human milk proresolving mediators stimulate resolution of acute inflammation.

PubMed

Arnardottir, Hildur; Orr, Sarah K; Dalli, Jesmond; Serhan, Charles N

2016-05-01

Human milk contains nutrients and bioactive products relevant to infant development and immunological protection. Here, we investigated the proresolving properties of milk using human milk lipid mediator isolates (HLMIs) and determined their impact on resolution programs in vivo and with human macrophages. HLMIs reduced the maximum neutrophil numbers (14.6±1.2 × 10(6)-11.0±1.0 × 10(6) cells per exudate) and shortened the resolution interval (Ri; 50% neutrophil reduction) by 54% compared with peritonitis. Using rigorous liquid-chromatography tandem-mass spectrometry (LC-MS-MS)-based lipid mediator (LM) metabololipidomics, we demonstrated that human milk possesses a proresolving LM-specialized proresolving mediator (LM-SPM) signature profile, containing SPMs (e.g. resolvins (Rv), protectins (PDs), maresins (MaRs), and lipoxins (LXs)) at bioactive levels (pico-nanomolar concentrations) that enhanced human macrophage efferocytosis and bacterial containment. SPMs identified in human milk included D-series Rvs (e.g., RvD1, RvD2, RvD3, AT-RvD3, and RvD4), PD1, MaR1, E-series Rvs (e.g. RvE1, RvE2, and RvE3), and LXs (LXA4 and LXB4). Of the SPMs identified in human milk, RvD2 and MaR1 (50 ng per mouse) individually shortened Ri by ∼75%. Milk from mastitis gave higher leukotriene B4 and prostanoids and lower SPM levels. Taken together, these findings provide evidence that human milk has proresolving actions via comprehensive LM-SPM profiling, describing a potentially novel mechanism in maternal-infant biochemical imprinting.

Tissuelike 3D Assemblies of Human Broncho-Epithelial Cells

NASA Technical Reports Server (NTRS)

Goodwin, Thomas J.

2010-01-01

Three-dimensional (3D) tissuelike assemblies (TLAs) of human broncho-epithelial (HBE) cells have been developed for use in in vitro research on infection of humans by respiratory viruses. The 2D monolayer HBE cell cultures heretofore used in such research lack the complex cell structures and interactions characteristic of in vivo tissues and, consequently, do not adequately emulate the infection dynamics of in-vivo microbial adhesion and invasion. In contrast, the 3D HBE TLAs are characterized by more-realistic reproductions of the geometrical and functional complexity, differentiation of cells, cell-to-cell interactions, and cell-to-matrix interactions characteristic of human respiratory epithelia. Hence, the 3D HBE TLAs are expected to make it possible to perform at least some of the research in vitro under more-realistic conditions, without need to infect human subjects. The TLAs are grown on collagen-coated cyclodextran microbeads under controlled conditions in a nutrient liquid in the simulated microgravitational environment of a bioreactor of the rotating- wall-vessel type. Primary human mesenchymal bronchial-tracheal cells are used as a foundation matrix, while adult human bronchial epithelial immortalized cells are used as the overlying component. The beads become coated with cells, and cells on adjacent beads coalesce into 3D masses. The resulting TLAs have been found to share significant characteristics with in vivo human respiratory epithelia including polarization, tight junctions, desmosomes, and microvilli. The differentiation of the cells in these TLAs into tissues functionally similar to in vivo tissues is confirmed by the presence of compounds, including villin, keratins, and specific lung epithelium marker compounds, and by the production of tissue mucin. In a series of initial infection tests, TLA cultures were inoculated with human respiratory syncytial viruses and parainfluenza type 3 viruses. Infection was confirmed by photomicrographs that showed signs of damage by viruses and virus titers (see figure) that indicated large increases in the populations of viruses during the days following inoculation.

Generation and use of human 3D-CAD models

NASA Astrophysics Data System (ADS)

Grotepass, Juergen; Speyer, Hartmut; Kaiser, Ralf

2002-05-01

Individualized Products are one of the ten mega trends of the 21st Century with human modeling as the key issue for tomorrow's design and product development. The use of human modeling software for computer based ergonomic simulations within the production process increases quality while reducing costs by 30- 50 percent and shortening production time. This presentation focuses on the use of human 3D-CAD models for both, the ergonomic design of working environments and made to measure garment production. Today, the entire production chain can be designed, individualized models generated and analyzed in 3D computer environments. Anthropometric design for ergonomics is matched to human needs, thus preserving health. Ergonomic simulation includes topics as human vision, reachability, kinematics, force and comfort analysis and international design capabilities. In German more than 17 billions of Mark are moved to other industries, because clothes do not fit. Individual clothing tailored to the customer's preference means surplus value, pleasure and perfect fit. The body scanning technology is the key to generation and use of human 3D-CAD models for both, the ergonomic design of working environments and made to measure garment production.

3-D ultrafast Doppler imaging applied to the noninvasive mapping of blood vessels in vivo.

PubMed

Provost, Jean; Papadacci, Clement; Demene, Charlie; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

2015-08-01

Ultrafast Doppler imaging was introduced as a technique to quantify blood flow in an entire 2-D field of view, expanding the field of application of ultrasound imaging to the highly sensitive anatomical and functional mapping of blood vessels. We have recently developed 3-D ultrafast ultrasound imaging, a technique that can produce thousands of ultrasound volumes per second, based on a 3-D plane and diverging wave emissions, and demonstrated its clinical feasibility in human subjects in vivo. In this study, we show that noninvasive 3-D ultrafast power Doppler, pulsed Doppler, and color Doppler imaging can be used to perform imaging of blood vessels in humans when using coherent compounding of 3-D tilted plane waves. A customized, programmable, 1024-channel ultrasound system was designed to perform 3-D ultrafast imaging. Using a 32 × 32, 3-MHz matrix phased array (Vermon, Tours, France), volumes were beamformed by coherently compounding successive tilted plane wave emissions. Doppler processing was then applied in a voxel-wise fashion. The proof of principle of 3-D ultrafast power Doppler imaging was first performed by imaging Tygon tubes of various diameters, and in vivo feasibility was demonstrated by imaging small vessels in the human thyroid. Simultaneous 3-D color and pulsed Doppler imaging using compounded emissions were also applied in the carotid artery and the jugular vein in one healthy volunteer.

Personalized development of human organs using 3D printing technology.

PubMed

Radenkovic, Dina; Solouk, Atefeh; Seifalian, Alexander

2016-02-01

3D printing is a technique of fabricating physical models from a 3D volumetric digital image. The image is sliced and printed using a specific material into thin layers, and successive layering of the material produces a 3D model. It has already been used for printing surgical models for preoperative planning and in constructing personalized prostheses for patients. The ultimate goal is to achieve the development of functional human organs and tissues, to overcome limitations of organ transplantation created by the lack of organ donors and life-long immunosuppression. We hypothesized a precision medicine approach to human organ fabrication using 3D printed technology, in which the digital volumetric data would be collected by imaging of a patient, i.e. CT or MRI images followed by mathematical modeling to create a digital 3D image. Then a suitable biocompatible material, with an optimal resolution for cells seeding and maintenance of cell viability during the printing process, would be printed with a compatible printer type and finally implanted into the patient. Life-saving operations with 3D printed implants were already performed in patients. However, several issues need to be addressed before translational application of 3D printing into clinical medicine. These are vascularization, innervation, and financial cost of 3D printing and safety of biomaterials used for the construct. Copyright © 2015 Elsevier Ltd. All rights reserved.

Introducing 3-Dimensional Printing of a Human Anatomic Pathology Specimen: Potential Benefits for Undergraduate and Postgraduate Education and Anatomic Pathology Practice.

PubMed

Mahmoud, Amr; Bennett, Michael

2015-08-01

Three-dimensional (3D) printing, a rapidly advancing technology, is widely applied in fields such as mechanical engineering and architecture. Three-dimensional printing has been introduced recently into medical practice in areas such as reconstructive surgery, as well as in clinical research. Three-dimensionally printed models of anatomic and autopsy pathology specimens can be used for demonstrating pathology entities to undergraduate medical, dental, and biomedical students, as well as for postgraduate training in examination of gross specimens for anatomic pathology residents and pathology assistants, aiding clinicopathological correlation at multidisciplinary team meetings, and guiding reconstructive surgical procedures. To apply 3D printing in anatomic pathology for teaching, training, and clinical correlation purposes. Multicolored 3D printing of human anatomic pathology specimens was achieved using a ZCorp 510 3D printer (3D Systems, Rock Hill, South Carolina) following creation of a 3D model using Autodesk 123D Catch software (Autodesk, Inc, San Francisco, California). Three-dimensionally printed models of anatomic pathology specimens created included pancreatoduodenectomy (Whipple operation) and radical nephrectomy specimens. The models accurately depicted the topographic anatomy of selected specimens and illustrated the anatomic relation of excised lesions to adjacent normal tissues. Three-dimensional printing of human anatomic pathology specimens is achievable. Advances in 3D printing technology may further improve the quality of 3D printable anatomic pathology specimens.

3D-fibroblast tissues constructed by a cell-coat technology enhance tight-junction formation of human colon epithelial cells.

PubMed

Matsusaki, Michiya; Hikimoto, Daichi; Nishiguchi, Akihiro; Kadowaki, Koji; Ohura, Kayoko; Imai, Teruko; Akashi, Mitsuru

2015-02-13

Caco-2, human colon carcinoma cell line, has been widely used as a model system for intestinal epithelial permeability because Caco-2 cells express tight-junctions, microvilli, and a number of enzymes and transporters characteristic of enterocytes. However, the functional differentiation and polarization of Caco-2 cells to express sufficient tight-junctions (a barrier) usually takes over 21 days in culture. This may be due to the cell culture environment, for example inflammation induced by plastic petri dishes. Three-dimensional (3D) sufficient cell microenvironments similar to in vivo natural conditions (proteins and cells), will promote rapid differentiation and higher functional expression of tight junctions. Herein we report for the first time an enhancement in tight-junction formation by 3D-cultures of Caco-2 cells on monolayered (1L) and eight layered (8L) normal human dermal fibroblasts (NHDF). Trans epithelial electric resistance (TEER) of Caco-2 cells was enhanced in the 3D-cultures, especially 8L-NHDF tissues, depending on culture times and only 10 days was enough to reach the same TEER value of Caco-2 monolayers after a 21 day incubation. Relative mRNA expression of tight-junction proteins of Caco-2 cells on 3D-cultures showed higher values than those in monolayer structures. Transporter gene expression patterns of Caco-2 cells on 3D-constructs were almost the same as those of Caco-2 monolayers, suggesting that there was no effect of 3D-cultures on transporter protein expression. The expression correlation between carboxylesterase 1 and 2 in 3D-cultures represented similar trends with human small intestines. The results of this study clearly represent a valuable application of 3D-Caco-2 tissues for pharmaceutical applications. Copyright © 2015 Elsevier Inc. All rights reserved.

Evaluation of stereoscopic display with visual function and interview

NASA Astrophysics Data System (ADS)

Okuyama, Fumio

1999-05-01

The influence of binocular stereoscopic (3D) television display on the human eye were compared with one of a 2D display, using human visual function testing and interviews. A 40- inch double lenticular display was used for 2D/3D comparison experiments. Subjects observed the display for 30 minutes at a distance 1.0 m, with a combination of 2D material and one of 3D material. The participants were twelve young adults. Main optometric test with visual function measured were visual acuity, refraction, phoria, near vision point, accommodation etc. The interview consisted of 17 questions. Testing procedures were performed just before watching, just after watching, and forty-five minutes after watching. Changes in visual function are characterized as prolongation of near vision point, decrease of accommodation and increase in phoria. 3D viewing interview results show much more visual fatigue in comparison with 2D results. The conclusions are: 1) change in visual function is larger and visual fatigue is more intense when viewing 3D images. 2) The evaluation method with visual function and interview proved to be very satisfactory for analyzing the influence of stereoscopic display on human eye.

Enhancement of 5-keto-d-gluconate production by a recombinant Gluconobacter oxydans using a dissolved oxygen control strategy.

PubMed

Yuan, Jianfeng; Wu, Mianbin; Lin, Jianping; Yang, Lirong

2016-07-01

The rapid and incomplete oxidation of sugars, alcohols, and polyols by the gram-negative bacterium Gluconobacter oxydans facilitates a wide variety of biological applications. For the conversion of glucose to 5-keto-d-gluconate (5-KGA), a promising precursor of the industrial substance L-(+)-tartaric acid, G. oxydans DSM2343 was genetically engineered to strain ZJU2, in which the GOX1231 and GOX1081 genes were knocked out in a markerless fashion. Then, a secondary alcohol dehydrogenase (GCD) from Xanthomonas campestris DSM3586 was heterologously expressed in G. oxydans ZJU2. The 5-KGA production and cell yield were increased by 10% and 24.5%, respectively. The specific activity of GCD towards gluconate was 1.75±0.02 U/mg protein, which was 7-fold higher than that of the sldAB in G. oxydans. Based on the analysis of kinetic parameters including specific cell growth rate (μ), specific glucose consumption rate (qs) and specific 5-KGA production rate (qp), a dissolved oxygen (DO) control strategy was proposed. Finally, batch fermentation was carried out in a 15-L bioreactor using an initial agitation speed of 600 rpm to obtain a high μ for cell growth. Subsequently, DO was continuously maintained above 20% to achieve a high qp to ensure a high accumulation of 5-KGA. Under these conditions, the maximum concentration of 5-KGA reached 117.75 g/L with a productivity of 2.10 g/(L·h). Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Fly versus man: evolutionary impairment of nucleolar targeting affects the degradome of Drosophila's Taspase1.

PubMed

Wünsch, Désirée; Hahlbrock, Angelina; Heiselmayer, Christina; Bäcker, Sandra; Heun, Patrick; Goesswein, Dorothee; Stöcker, Walter; Schirmeister, Tanja; Schneider, Günter; Krämer, Oliver H; Knauer, Shirley K; Stauber, Roland H

2015-05-01

Human Taspase1 is essential for development and cancer by processing critical regulators, such as the mixed-lineage leukemia protein. Likewise, its ortholog, trithorax, is cleaved by Drosophila Taspase1 (dTaspase1), implementing a functional coevolution. To uncover novel mechanism regulating protease function, we performed a functional analysis of dTaspase1 and its comparison to the human ortholog. dTaspase1 contains an essential nucleophile threonine(195), catalyzing cis cleavage into its α- and β-subunits. A cell-based assay combined with alanine scanning mutagenesis demonstrated that the target cleavage motif for dTaspase1 (Q(3)[F/I/L/M](2)D(1)↓G(1')X(2')X(3')) differs significantly from the human ortholog (Q(3)[F,I,L,V](2)D(1)↓G(1')x(2')D(3')D(4')), predicting an enlarged degradome containing 70 substrates for Drosophila. In contrast to human Taspase1, dTaspase1 shows no discrete localization to the nucleus/nucleolus due to the lack of the importin-α/nucleophosmin1 interaction domain (NoLS) conserved in all vertebrates. Consequently, dTaspase1 interacts with neither the Drosophila nucleoplasmin-like protein nor human nucleophosmin1. The impact of localization on the protease's degradome was confirmed by demonstrating that dTaspase1 did not efficiently process nuclear substrates, such as upstream stimulatory factor 2. However, genetic introduction of the NoLS into dTaspase1 restored its nucleolar localization, nucleophosmin1 interaction, and efficient cleavage of nuclear substrates. We report that evolutionary functional divergence separating vertebrates from invertebrates can be achieved for proteases by a transport/localization-regulated mechanism. © FASEB.

Imaging dopamine D3 receptors in the human brain with positron emission tomography, [11C]PHNO, and a selective D3 receptor antagonist.

PubMed

Searle, Graham; Beaver, John D; Comley, Robert A; Bani, Massimo; Tziortzi, Andri; Slifstein, Mark; Mugnaini, Manolo; Griffante, Cristiana; Wilson, Alan A; Merlo-Pich, Emilio; Houle, Sylvain; Gunn, Roger; Rabiner, Eugenii A; Laruelle, Marc

2010-08-15

Dopamine D(3) receptors are involved in the pathophysiology of several neuropsychiatric conditions. [(11)C]-(+)-PHNO is a radiolabeled D(2) and D(3) agonist, suitable for imaging the agonist binding sites (denoted D(2HIGH) and D(3)) of these receptors with positron emission tomography (PET). PET studies in nonhuman primates documented that, in vivo, [(11)C]-(+)-PHNO displays a relative selectivity for D(3) compared with D(2HIGH) receptor sites and that the [(11)C]-(+)-PHNO signal is enriched in D(3) contribution compared with conventional ligands such as [(11)C] raclopride. To define the D(3) contribution (f(PHNO)(D3)) to [(11)C]-(+)-PHNO binding potential (BP(ND)) in healthy humans, 52 PET scans were obtained in 19 healthy volunteers at baseline and following oral administration of various doses of the selective D(3) receptor antagonist, GSK598809. The impact of GSK598809 on [(11)C]-(+)-PHNO was regionally selective. In dorsal regions of the striatum, GSK598809 did not significantly affect [(11)C]-(+)-PHNO BP(ND) (f(PHNO)(D3) approximately 0%). Conversely, in the substantia nigra, GSK598809 dose-dependently reduced [(11)C]-(+)-PHNO binding to nonspecific level (f(PHNO)(D3) approximately 100%). In ventral striatum (VST), globus pallidus and thalamus (THA), [(11)C]-(+)-PHNO BP(ND) was attributable to a combination of D(2HIGH) and D(3) receptor sites, with f(PHNO)(D3) of 26%, 67% and 46%, respectively. D(3) receptor binding potential (BP(ND)(D3)) was highest in globus pallidus (1.90) and substantial nigra (1.39), with lower levels in VST (.77) and THA (.18) and negligible levels in dorsal striatum. This study elucidated the pharmacologic nature of the [(11)C]-(+)-PHNO signal in healthy subjects and provided the first quantification of D(3) receptor availability with PET in the living human brain. Copyright 2010 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.

Hepatic differentiation of human iPSCs in different 3D models: A comparative study

PubMed Central

Brzeszczynska, Joanna; Knöspel, Fanny; Armstrong, Lyle; Lako, Majlinda; Greuel, Selina; Damm, Georg; Ludwig-Schwellinger, Eva; Deschl, Ulrich; Ross, James A.

2017-01-01

Human induced pluripotent stem cells (hiPSCs) are a promising source from which to derive distinct somatic cell types for in vitro or clinical use. Existent protocols for hepatic differentiation of hiPSCs are primarily based on 2D cultivation of the cells. In the present study, the authors investigated the generation of hiPSC-derived hepatocyte-like cells using two different 3D culture systems: A 3D scaffold-free microspheroid culture system and a 3D hollow-fiber perfusion bioreactor. The differentiation outcome in these 3D systems was compared with that in conventional 2D cultures, using primary human hepatocytes as a control. The evaluation was made based on specific mRNA expression, protein secretion, antigen expression and metabolic activity. The expression of α-fetoprotein was lower, while cytochrome P450 1A2 or 3A4 activities were higher in the 3D culture systems as compared with the 2D differentiation system. Cells differentiated in the 3D bioreactor showed an increased expression of albumin and hepatocyte nuclear factor 4α, as well as secretion of α-1-antitrypsin as compared with the 2D differentiation system, suggesting a higher degree of maturation. In contrast, the 3D scaffold-free microspheroid culture provides an easy and robust method to generate spheroids of a defined size for screening applications, while the bioreactor culture model provides an instrument for complex investigations under physiological-like conditions. In conclusion, the present study introduces two 3D culture systems for stem cell derived hepatic differentiation each demonstrating advantages for individual applications as well as benefits in comparison with 2D cultures. PMID:29039463

In vitro enteroid-derived three-dimensional tissue model of human small intestinal epithelium with innate immune responses.

PubMed

Chen, Ying; Zhou, Wenda; Roh, Terrence; Estes, Mary K; Kaplan, David L

2017-01-01

There is a need for functional in vitro 3D human intestine systems that can bridge the gap between conventional cell culture studies and human trials. The successful engineering in vitro of human intestinal tissues relies on the use of the appropriate cell sources, biomimetic scaffolds, and 3D culture conditions to support vital organ functions. We previously established a compartmentalized scaffold consisting of a hollow space within a porous bulk matrix, in which a functional and physiologically relevant intestinal epithelium system was generated using intestinal cell lines. In this study, we adopt the 3D scaffold system for the cultivation of stem cell-derived human small intestinal enteriods (HIEs) to engineer an in vitro 3D model of a nonstransformed human small intestinal epithelium. Characterization of tissue properties revealed a mature HIE-derived epithelium displaying four major terminally differentiated epithelial cell types (enterocytes, Goblet cells, Paneth cells, enteroendocrine cells), with tight junction formation, microvilli polarization, digestive enzyme secretion, and low oxygen tension in the lumen. Moreover, the tissue model demonstrates significant antibacterial responses to E. coli infection, as evidenced by the significant upregulation of genes involved in the innate immune response. Importantly, many of these genes are activated in human patients with inflammatory bowel disease (IBD), implicating the potential application of the 3D stem-cell derived epithelium for the in vitro study of host-microbe-pathogen interplay and IBD pathogenesis.

Downregulation of tight junction-associated MARVEL protein marvelD3 during epithelial-mesenchymal transition in human pancreatic cancer cells.

PubMed

Kojima, Takashi; Takasawa, Akira; Kyuno, Daisuke; Ito, Tatsuya; Yamaguchi, Hiroshi; Hirata, Koichi; Tsujiwaki, Mitsuhiro; Murata, Masaki; Tanaka, Satoshi; Sawada, Norimasa

2011-10-01

The novel tight junction protein marvelD3 contains a conserved MARVEL (MAL and related proteins for vesicle trafficking and membrane link) domain like occludin and tricellulin. However, little is yet known about the detailed role and regulation of marvelD3 in normal epithelial cells and cancer cells, including pancreatic cancer. In the present study, we investigated marvelD3 expression in well and poorly differentiated human pancreatic cancer cell lines and normal pancreatic duct epithelial cells in which the hTERT gene was introduced into human pancreatic duct epithelial cells in primary culture, and the changes of marvelD3 during Snail-induced epithelial-mesenchymal transition (EMT) under hypoxia, TGF-β treatment and knockdown of FOXA2 in well differentiated pancreatic cancer HPAC cells. MarvelD3 was transcriptionally downregulated in poorly differentiated pancreatic cancer cells and during Snail-induced EMT of pancreatic cancer cells in which Snail was highly expressed and the fence function downregulated, whereas it was maintained in well differentiated human pancreatic cancer cells and normal pancreatic duct epithelial cells. Depletion of marvelD3 by siRNAs in HPAC cells resulted in downregulation of barrier functions indicated as a decrease in transepithelial electric resistance and an increase of permeability to fluorescent dextran tracers, whereas it did not affect fence function of tight junctions. In conclusion, marvelD3 is transcriptionally downregulated in Snail-induced EMT during the progression for the pancreatic cancer. Copyright © 2011 Elsevier Inc. All rights reserved.

Functional metabolic interactions of human neuron-astrocyte 3D in vitro networks

PubMed Central

Simão, Daniel; Terrasso, Ana P.; Teixeira, Ana P.; Brito, Catarina; Sonnewald, Ursula; Alves, Paula M.

2016-01-01

The generation of human neural tissue-like 3D structures holds great promise for disease modeling, drug discovery and regenerative medicine strategies. Promoting the establishment of complex cell-cell interactions, 3D culture systems enable the development of human cell-based models with increased physiological relevance, over monolayer cultures. Here, we demonstrate the establishment of neuronal and astrocytic metabolic signatures and shuttles in a human 3D neural cell model, namely the glutamine-glutamate-GABA shuttle. This was indicated by labeling of neuronal GABA following incubation with the glia-specific substrate [2-13C]acetate, which decreased by methionine sulfoximine-induced inhibition of the glial enzyme glutamine synthetase. Cell metabolic specialization was further demonstrated by higher pyruvate carboxylase-derived labeling in glutamine than in glutamate, indicating its activity in astrocytes and not in neurons. Exposure to the neurotoxin acrylamide resulted in intracellular accumulation of glutamate and decreased GABA synthesis. These results suggest an acrylamide-induced impairment of neuronal synaptic vesicle trafficking and imbalanced glutamine-glutamate-GABA cycle, due to loss of cell-cell contacts at synaptic sites. This work demonstrates, for the first time to our knowledge, that neural differentiation of human cells in a 3D setting recapitulates neuronal-astrocytic metabolic interactions, highlighting the relevance of these models for toxicology and better understanding the crosstalk between human neural cells. PMID:27619889

Functional metabolic interactions of human neuron-astrocyte 3D in vitro networks.

PubMed

Simão, Daniel; Terrasso, Ana P; Teixeira, Ana P; Brito, Catarina; Sonnewald, Ursula; Alves, Paula M

2016-09-13

The generation of human neural tissue-like 3D structures holds great promise for disease modeling, drug discovery and regenerative medicine strategies. Promoting the establishment of complex cell-cell interactions, 3D culture systems enable the development of human cell-based models with increased physiological relevance, over monolayer cultures. Here, we demonstrate the establishment of neuronal and astrocytic metabolic signatures and shuttles in a human 3D neural cell model, namely the glutamine-glutamate-GABA shuttle. This was indicated by labeling of neuronal GABA following incubation with the glia-specific substrate [2-(13)C]acetate, which decreased by methionine sulfoximine-induced inhibition of the glial enzyme glutamine synthetase. Cell metabolic specialization was further demonstrated by higher pyruvate carboxylase-derived labeling in glutamine than in glutamate, indicating its activity in astrocytes and not in neurons. Exposure to the neurotoxin acrylamide resulted in intracellular accumulation of glutamate and decreased GABA synthesis. These results suggest an acrylamide-induced impairment of neuronal synaptic vesicle trafficking and imbalanced glutamine-glutamate-GABA cycle, due to loss of cell-cell contacts at synaptic sites. This work demonstrates, for the first time to our knowledge, that neural differentiation of human cells in a 3D setting recapitulates neuronal-astrocytic metabolic interactions, highlighting the relevance of these models for toxicology and better understanding the crosstalk between human neural cells.

Synthesis of calbindin-D28K during mineralization in human bone marrow stromal cells.

PubMed Central

Faucheux, C; Bareille, R; Amedee, J

1998-01-01

1alpha,25-Dihydroxyvitamin D3 [1,25(OH)2D3] is known to modulate Ca2+ metabolism in several cell types. Vitamin-D-dependent calcium binding proteins such as calbindin-D28K (28 kDa calcium binding proteins) have been shown to be regulated by 1,25(OH)2D3 but the mechanisms controlling calbindin synthesis are still poorly understood in human osteoblast cell culture models. The human bone marrow stromal cells (HBMSC) described in this paper developed a calcified matrix, expressed osteocalcin (OC), osteopontin (OP) and responded to 1,25(OH)2D3. The expression of vitamin D receptor mRNA was demonstrated by reverse transcription-PCR. Calbindin-D28K protein was identified only in cells arising from the sixth subculture, which exhibited a calcified matrix and all of the osteoblastic markers, e.g. OC and OP. It was demonstrated by dot-immunodetection using immunological probes, and by in situ hybridization using labelled cDNA probes. Moreover, vitamin D3 enhanced calbindin-D28K synthesis as well as OC synthesis and alkaline phosphatase activity. Uptake of 45Ca induced into the matrix by 1,25(OH)2D3 supports the hypothesis that the calcium-enriched matrix could trap calbindin-D proteins. In conclusion, the studies in vitro described in the present paper indicate, for the first time, a possible role of calbindin-D28K in mineralized matrix formation in HBMSC. PMID:9677345

Human Body 3D Posture Estimation Using Significant Points and Two Cameras

PubMed Central

Juang, Chia-Feng; Chen, Teng-Chang; Du, Wei-Chin

2014-01-01

This paper proposes a three-dimensional (3D) human posture estimation system that locates 3D significant body points based on 2D body contours extracted from two cameras without using any depth sensors. The 3D significant body points that are located by this system include the head, the center of the body, the tips of the feet, the tips of the hands, the elbows, and the knees. First, a linear support vector machine- (SVM-) based segmentation method is proposed to distinguish the human body from the background in red, green, and blue (RGB) color space. The SVM-based segmentation method uses not only normalized color differences but also included angle between pixels in the current frame and the background in order to reduce shadow influence. After segmentation, 2D significant points in each of the two extracted images are located. A significant point volume matching (SPVM) method is then proposed to reconstruct the 3D significant body point locations by using 2D posture estimation results. Experimental results show that the proposed SVM-based segmentation method shows better performance than other gray level- and RGB-based segmentation approaches. This paper also shows the effectiveness of the 3D posture estimation results in different postures. PMID:24883422

Cytochrome P450 dependent metabolism of the new designer drug 1-(3-trifluoromethylphenyl)piperazine (TFMPP). In vivo studies in Wistar and Dark Agouti rats as well as in vitro studies in human liver microsomes.

PubMed

Staack, Roland F; Paul, Liane D; Springer, Dietmar; Kraemer, Thomas; Maurer, Hans H

2004-01-15

1-(3-Trifluoromethylphenyl)piperazine (TFMPP) is a designer drug with serotonergic properties. Previous studies with male Wistar rats (WI) had shown, that TFMPP was metabolized mainly by aromatic hydroxylation. In the current study, it was examined whether this reaction may be catalyzed by cytochrome P450 (CYP)2D6 by comparing TFMPP vs. hydroxy TFMPP ratios in urine from female Dark Agouti rats, a model of the human CYP2D6 poor metabolizer phenotype (PM), male Dark Agouti rats, an intermediate model, and WI, a model of the human CYP2D6 extensive metabolizer phenotype. Furthermore, the human hepatic CYPs involved in TFMPP hydroxylation were identified using cDNA-expressed CYPs and human liver microsomes. Finally, TFMPP plasma levels in the above mentioned rats were compared. The urine studies suggested that TFMPP hydroxylation might be catalyzed by CYP2D6 in humans. Studies using human CYPs showed that CYP1A2, CYP2D6 and CYP3A4 catalyzed TFMPP hydroxylation, with CYP2D6 being the most important enzyme accounting for about 81% of the net intrinsic clearance, calculated using the relative activity factor approach. The hydroxylation was significantly inhibited by quinidine (77%) and metabolite formation in poor metabolizer genotype human liver microsomes was significantly lower (63%) compared to pooled human liver microsomes. Analysis of the plasma samples showed that female Dark Agouti rats exhibited significantly higher TFMPP plasma levels compared to those of male Dark Agouti rats and WI. Furthermore, pretreatment of WI with the CYP2D inhibitor quinine resulted in significantly higher TFMPP plasma levels. In conclusion, the presented data give hints for possible differences in pharmacokinetics in human PM and human CYP2D6 extensive metabolizer phenotype subjects relevant for risk assessment.

Basis of Miscoding of the DNA Adduct N2,3-Ethenoguanine by Human Y-family DNA Polymerases*

PubMed Central

Zhao, Linlin; Pence, Matthew G.; Christov, Plamen P.; Wawrzak, Zdzislaw; Choi, Jeong-Yun; Rizzo, Carmelo J.; Egli, Martin; Guengerich, F. Peter

2012-01-01

N2,3-Ethenoguanine (N2,3-ϵG) is one of the exocyclic DNA adducts produced by endogenous processes (e.g. lipid peroxidation) and exposure to bioactivated vinyl monomers such as vinyl chloride, which is a known human carcinogen. Existing studies exploring the miscoding potential of this lesion are quite indirect because of the lability of the glycosidic bond. We utilized a 2′-fluoro isostere approach to stabilize this lesion and synthesized oligonucleotides containing 2′-fluoro-N2,3-ϵ-2′-deoxyarabinoguanosine to investigate the miscoding potential of N2,3-ϵG by Y-family human DNA polymerases (pols). In primer extension assays, pol η and pol κ replicated through N2,3-ϵG, whereas pol ι and REV1 yielded only 1-base incorporation. Steady-state kinetics revealed that dCTP incorporation is preferred opposite N2,3-ϵG with relative efficiencies in the order of pol κ > REV1 > pol η ≈ pol ι, and dTTP misincorporation is the major miscoding event by all four Y-family human DNA pols. Pol ι had the highest dTTP misincorporation frequency (0.71) followed by pol η (0.63). REV1 misincorporated dTTP and dGTP with much lower frequencies. Crystal structures of pol ι with N2,3-ϵG paired to dCTP and dTTP revealed Hoogsteen-like base pairing mechanisms. Two hydrogen bonds were observed in the N2,3-ϵG:dCTP base pair, whereas only one appears to be present in the case of the N2,3-ϵG:dTTP pair. Base pairing mechanisms derived from the crystal structures explain the slightly favored dCTP insertion for pol ι in steady-state kinetic analysis. Taken together, these results provide a basis for the mutagenic potential of N2,3-ϵG. PMID:22910910

Conditioned medium from the three-dimensional culture of human umbilical cord perivascular cells accelerate the migration and proliferation of human keratinocyte and fibroblast.

PubMed

Kim, Min Ho; Wu, Wen Hao; Choi, Jee Hyun; Kim, Ji Hyun; Hong, Seok-Ho; Jun, Jin Hyun; Ko, Yong; Lee, Jong Hun

Previous studies have reported that the conditioned medium (CM) of bone marrow-mesenchymal stem cells (BM-MSCs) stimulate the migration and proliferation of cell types involved in the wound healing process. However, these studies only show MSC-CM effects that were obtained using a two-dimensional (2D) culture. Recently, a three-dimensional (3D) culture has been considered to be a more physiologically appropriate system than the 2D culture. In addition, it has been shown that the procurement of BM-MSC is invasive, and other sources of MSC are thus being explored. Recently, perivascular cells (PVCs) have been considered as an alternative source of cells for dermal wound healing. Therefore, in this study, a PVC-conditioned medium (CM) was collected from a 3D culture (PVC-CM-3D) using highly porous polystyrene-based membranes and compared with PVC-CM from a 2D culture (PVC-CM-2D) to investigate the effects on the migration and proliferation of human keratinocytes and fibroblasts. Moreover, the PVC-CM components from the 2D and 3D cultures were identified using 2D gel electrophoresis. The migrations of the keratinocytes cells and fibroblasts were significantly higher with PVC-CM-3D than with the 2D culture; similarly, the proliferation of keratinocytes was also highly stimulated by PVC-CM-3D. Proteomic analyses of the PVC-CM revealed that type I collagen was highly expressed in the 3D-culture system. Microtubule-actin cross-linked factor 1 (KIAA0465), nebulin-related anchoring protein, and thioredoxin were specifically expressed only in PVC-CM-3D. In addition, more EVs could be isolated from the PVC-CM-3D, and EVs were found to stimulate keratinocyte migration. Taken together, 3D-culture using a polystyrene scaffold is demonstrated to be a better system for providing better physiological conditions; therefore, PVC-CM-3D could be a promising option for skin-wound healing.

Downregulation of Ubiquitin-conjugating Enzyme UBE2D3 Promotes Telomere Maintenance and Radioresistance of Eca-109 Human Esophageal Carcinoma Cells

PubMed Central

Yang, Hui; Wu, Lin; Ke, Shaobo; Wang, Wenbo; Yang, Lei; Gao, Xiaojia; Fang, Hongyan; Yu, Haijun; Zhong, Yahua; Xie, Conghua; Zhou, Fuxiang; Zhou, Yunfeng

2016-01-01

Ubiquitin-conjugating enzyme UBE2D3 is an important member of the ubiquitin-proteasome pathways. Our previous study showed that the expression of UBE2D3 was negatively related to human telomerase reverse transcriptase (hTERT) and radioresistance in human breast cancer cells. However, in esophageal carcinoma, the exact effects and mechanisms of UBE2D3 in radioresistance remain unclear. This study shows that UBE2D3 knockdown was associated with significant increases in radioresistance to X-rays, telomerase activity, telomere length, and telomere shelterins. UBE2D3 knockdown-mediated radioresistance was related to a decrease in the spontaneous and ionizing radiation-induced apoptosis, resulting from a decrease in the Bax/Bcl-2 ratio. Furthermore, UBE2D3 downregulation was associated with increased G1-S phase transition and prolonged IR-induced G2/M arrest through over expression of cyclin D1, decrease of CDC25A expression and promotion of the ATM/ATR-Chk1-CDC25C pathway. Moreover, UBE2D3 downregulation reduced spontaneous DNA double-strand breaks and accelerated the repair of DNA damage induced by IR. The current data thus demonstrate that UBE2D3 downregulation enhances radioresistance by increased telomere homeostasis and prolonged IR-induced G2/M arrest, but decreases the IR-induced apoptosis and the number of DNA damage foci. These results suggest that UBE2D3 might be a potential molecular target to improve radiotherapy effects in esophageal carcinoma. PMID:27326259

Engineering Human Neural Tissue by 3D Bioprinting.

PubMed

Gu, Qi; Tomaskovic-Crook, Eva; Wallace, Gordon G; Crook, Jeremy M

2018-01-01

Bioprinting provides an opportunity to produce three-dimensional (3D) tissues for biomedical research and translational drug discovery, toxicology, and tissue replacement. Here we describe a method for fabricating human neural tissue by 3D printing human neural stem cells with a bioink, and subsequent gelation of the bioink for cell encapsulation, support, and differentiation to functional neurons and supporting neuroglia. The bioink uniquely comprises the polysaccharides alginate, water-soluble carboxymethyl-chitosan, and agarose. Importantly, the method could be adapted to fabricate neural and nonneural tissues from other cell types, with the potential to be applied for both research and clinical product development.

Calcium and 1,25-dihydroxyvitamin D3 modulate genes of immune and inflammatory pathways in the human colon: a human crossover trial.

PubMed

Protiva, Petr; Pendyala, Swaroop; Nelson, Celeste; Augenlicht, Leonard H; Lipkin, Martin; Holt, Peter R

2016-05-01

A high dietary calcium intake with adequate vitamin D status has been linked to lower colorectal cancer risk, but the mechanisms of these effects are poorly understood. The objective of this study was to elucidate the effects of a Western-style diet (WD) and supplemental calcium and/or 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on the colorectal mucosa. We conducted 2 crossover trials to define molecular pathways in the human colorectum altered by 1) a 4-wk WD supplemented with and without 2 g calcium carbonate/d and 2) a 4-wk WD supplemented with 1,25(OH)2D3 (0.5 μg/d) with or without 2 g calcium carbonate/d. The primary study endpoint was genome-wide gene expression in biopsy specimens of the rectosigmoid colonic mucosa. Serum and urinary calcium concentrations were also measured. Changes in urinary calcium accurately reflected calcium consumption. The WD induced modest upregulation of genes involved in inflammatory pathways, including interferon signaling, and calcium supplementation reversed these toward baseline. In contrast, supplementation of the WD with 1,25(OH)2D3 induced striking upregulation of genes involved in inflammation, immune response, extracellular matrix, and cell adhesion. Calcium supplementation largely abrogated these changes. Supplementing 1,25(OH)2D3 to a WD markedly upregulated genes in immune response and inflammation pathways, which were largely reversed by calcium supplementation. This study provides clinical trial evidence of global gene expression changes occurring in the human colorectum in response to calcium and 1,25(OH)2D3 intervention. One action of 1,25(OH)2D3 is to upregulate adaptive immunity. Calcium appears to modulate this effect, pointing to its biological interaction in the mucosa. This trial was registered at clinicaltrials.gov as NCT00298545 Trial protocol is available at http://clinicalstudies.rucares.org (protocol numbers PHO475 and PHO554). © 2016 American Society for Nutrition.

An Outdoor Navigation Platform with a 3D Scanner and Gyro-assisted Odometry

NASA Astrophysics Data System (ADS)

Yoshida, Tomoaki; Irie, Kiyoshi; Koyanagi, Eiji; Tomono, Masahiro

This paper proposes a light-weight navigation platform that consists of gyro-assisted odometry, a 3D laser scanner and map-based localization for human-scale robots. The gyro-assisted odometry provides highly accurate positioning only by dead-reckoning. The 3D laser scanner has a wide field of view and uniform measuring-point distribution. The map-based localization is robust and computationally inexpensive by utilizing a particle filter on a 2D grid map generated by projecting 3D points on to the ground. The system uses small and low-cost sensors, and can be applied to a variety of mobile robots in human-scale environments. Outdoor navigation experiments were conducted at the Tsukuba Challenge held in 2009 and 2010, which is an open proving ground for human-scale robots. Our robot successfully navigated the assigned 1-km courses in a fully autonomous mode multiple times.

Registration algorithm research for three dimensional medical image

NASA Astrophysics Data System (ADS)

Zhao, Jianping; Yang, Huamin; Ding, Ying

2008-03-01

The development of CT and MRI etc. technique offers the means by which we can research directly human internal structure. In clinic, usually various imaging results of a patient are combined for analysis. At present, in the most case, doctors make a diagnosis by observing some slice images of human body. As complexity and configuration diversity of the structure of human body organ, and as well unpredictiveness of focus location and configuration, it is difficult to imagine the cubic configuration of organs and their relationship from these 2D slices without corresponding specialty knowledge and practical experience. So it isn't satisfied with preferable requests of medical diagnosis that only aligning two 2D images to get one 2D slice image. As a result we need extend registration t problem to 3D image. As the quantity of 3D volume data are much more, it undoubtedly increases calculation quantity for aligning two 3D images accurately. It forces us to find some good methods that can achieve better effect on precision and satisfy the demand for time. So in this paper digitally reconstructed radiograph (DRR) image method is proposed to solve correlative problems. Ray tracking two 3D images and digitally reconstruct to create two 2D images, by aligning 2D data to realize to align 3D data.

Use of patient specific 3D printed neurovascular phantoms to evaluate the clinical utility of a high resolution x-ray imager

NASA Astrophysics Data System (ADS)

Setlur Nagesh, S. V.; Russ, M.; Ionita, C. N.; Bednarek, D.; Rudin, S.

2017-03-01

Modern 3D printing technology can fabricate vascular phantoms based on an actual human patient with a high degree of precision facilitating a realistic simulation environment for an intervention. We present two experimental setups using 3D printed patient-specific neurovasculature to simulate different disease anatomies. To simulate the human neurovasculature in the Circle of Willis, patient-based phantoms with aneurysms were 3D printed using a Objet Eden 260V printer. Anthropomorphic head phantoms and a human skull combined with acrylic plates simulated human head bone anatomy and x-ray attenuation. For dynamic studies the 3D printed phantom was connected to a pulsatile flow loop with the anthropomorphic phantom underneath. By combining different 3D printed phantoms and the anthropomorphic phantoms, different patient pathologies can be simulated. For static studies a 3D printed neurovascular phantom was embedded inside a human skull and used as a positional reference for treatment devices such as stents. To simulate tissue attenuation acrylic layers were added. Different combinations can simulate different patient treatment procedures. The Complementary-Metal-Oxide-Semiconductor (CMOS) based High Resolution Fluoroscope (HRF) with 75μm pixels offers an advantage over the state-of-the-art 200 μm pixel Flat Panel Detector (FPD) due to higher Nyquist frequency and better DQE performance. Whether this advantage is clinically useful during an actual clinical neurovascular intervention can be addressed by qualitatively evaluating images from a cohort of various cases performed using both detectors. The above-mentioned method can offer a realistic substitute for an actual clinical procedure. Also a large cohort of cases can be generated and used for a HRF clinical utility determination study.

Evaluation of the Leap Motion Controller during the performance of visually-guided upper limb movements.

PubMed

Niechwiej-Szwedo, Ewa; Gonzalez, David; Nouredanesh, Mina; Tung, James

2018-01-01

Kinematic analysis of upper limb reaching provides insight into the central nervous system control of movements. Until recently, kinematic examination of motor control has been limited to studies conducted in traditional research laboratories because motion capture equipment used for data collection is not easily portable and expensive. A recently developed markerless system, the Leap Motion Controller (LMC), is a portable and inexpensive tracking device that allows recording of 3D hand and finger position. The main goal of this study was to assess the concurrent reliability and validity of the LMC as compared to the Optotrak, a criterion-standard motion capture system, for measures of temporal accuracy and peak velocity during the performance of upper limb, visually-guided movements. In experiment 1, 14 participants executed aiming movements to visual targets presented on a computer monitor. Bland-Altman analysis was conducted to assess the validity and limits of agreement for measures of temporal accuracy (movement time, duration of deceleration interval), peak velocity, and spatial accuracy (endpoint accuracy). In addition, a one-sample t-test was used to test the hypothesis that the error difference between measures obtained from Optotrak and LMC is zero. In experiment 2, 15 participants performed a Fitts' type aiming task in order to assess whether the LMC is capable of assessing a well-known speed-accuracy trade-off relationship. Experiment 3 assessed the temporal coordination pattern during the performance of a sequence consisting of a reaching, grasping, and placement task in 15 participants. Results from the t-test showed that the error difference in temporal measures was significantly different from zero. Based on the results from the 3 experiments, the average temporal error in movement time was 40±44 ms, and the error in peak velocity was 0.024±0.103 m/s. The limits of agreement between the LMC and Optotrak for spatial accuracy measures ranged between 2-5 cm. Although the LMC system is a low-cost, highly portable system, which could facilitate collection of kinematic data outside of the traditional laboratory settings, the temporal and spatial errors may limit the use of the device in some settings.

Evaluation of the Leap Motion Controller during the performance of visually-guided upper limb movements

PubMed Central

Gonzalez, David; Nouredanesh, Mina; Tung, James

2018-01-01

Kinematic analysis of upper limb reaching provides insight into the central nervous system control of movements. Until recently, kinematic examination of motor control has been limited to studies conducted in traditional research laboratories because motion capture equipment used for data collection is not easily portable and expensive. A recently developed markerless system, the Leap Motion Controller (LMC), is a portable and inexpensive tracking device that allows recording of 3D hand and finger position. The main goal of this study was to assess the concurrent reliability and validity of the LMC as compared to the Optotrak, a criterion-standard motion capture system, for measures of temporal accuracy and peak velocity during the performance of upper limb, visually-guided movements. In experiment 1, 14 participants executed aiming movements to visual targets presented on a computer monitor. Bland-Altman analysis was conducted to assess the validity and limits of agreement for measures of temporal accuracy (movement time, duration of deceleration interval), peak velocity, and spatial accuracy (endpoint accuracy). In addition, a one-sample t-test was used to test the hypothesis that the error difference between measures obtained from Optotrak and LMC is zero. In experiment 2, 15 participants performed a Fitts’ type aiming task in order to assess whether the LMC is capable of assessing a well-known speed-accuracy trade-off relationship. Experiment 3 assessed the temporal coordination pattern during the performance of a sequence consisting of a reaching, grasping, and placement task in 15 participants. Results from the t-test showed that the error difference in temporal measures was significantly different from zero. Based on the results from the 3 experiments, the average temporal error in movement time was 40±44 ms, and the error in peak velocity was 0.024±0.103 m/s. The limits of agreement between the LMC and Optotrak for spatial accuracy measures ranged between 2–5 cm. Although the LMC system is a low-cost, highly portable system, which could facilitate collection of kinematic data outside of the traditional laboratory settings, the temporal and spatial errors may limit the use of the device in some settings. PMID:29529064

A perceptual map for gait symmetry quantification and pathology detection.

PubMed

Moevus, Antoine; Mignotte, Max; de Guise, Jacques A; Meunier, Jean

2015-10-29

The gait movement is an essential process of the human activity and the result of collaborative interactions between the neurological, articular and musculoskeletal systems, working efficiently together. This explains why gait analysis is important and increasingly used nowadays for the diagnosis of many different types (neurological, muscular, orthopedic, etc.) of diseases. This paper introduces a novel method to quickly visualize the different parts of the body related to an asymmetric movement in the human gait of a patient for daily clinical usage. The proposed gait analysis algorithm relies on the fact that the healthy walk has (temporally shift-invariant) symmetry properties in the coronal plane. The goal is to provide an inexpensive and easy-to-use method, exploiting an affordable consumer depth sensor, the Kinect, to measure the gait asymmetry and display results in a perceptual way. We propose a multi-dimensional scaling mapping using a temporally shift invariant distance, allowing us to efficiently visualize (in terms of perceptual color difference) the asymmetric body parts of the gait cycle of a subject. We also propose an index computed from this map and which quantifies locally and globally the degree of asymmetry. The proposed index is proved to be statistically significant and this new, inexpensive, marker-less, non-invasive, easy to set up, gait analysis system offers a readable and flexible tool for clinicians to analyze gait characteristics and to provide a fast diagnostic. This system, which estimates a perceptual color map providing a quick overview of asymmetry existing in the gait cycle of a subject, can be easily exploited for disease progression, recovery cues from post-operative surgery (e.g., to check the healing process or the effect of a treatment or a prosthesis) or might be used for other pathologies where gait asymmetry might be a symptom.

3D surface perception from motion involves a temporal–parietal network

PubMed Central

Beer, Anton L.; Watanabe, Takeo; Ni, Rui; Sasaki, Yuka; Andersen, George J.

2010-01-01

Previous research has suggested that three-dimensional (3D) structure-from-motion (SFM) perception in humans involves several motion-sensitive occipital and parietal brain areas. By contrast, SFM perception in nonhuman primates seems to involve the temporal lobe including areas MT, MST and FST. The present functional magnetic resonance imaging study compared several motion-sensitive regions of interest including the superior temporal sulcus (STS) while human observers viewed horizontally moving dots that defined either a 3D corrugated surface or a 3D random volume. Low-level stimulus features such as dot density and velocity vectors as well as attention were tightly controlled. Consistent with previous research we found that 3D corrugated surfaces elicited stronger responses than random motion in occipital and parietal brain areas including area V3A, the ventral and dorsal intraparietal sulcus, the lateral occipital sulcus and the fusiform gyrus. Additionally, 3D corrugated surfaces elicited stronger activity in area MT and the STS but not in area MST. Brain activity in the STS but not in area MT correlated with interindividual differences in 3D surface perception. Our findings suggest that area MT is involved in the analysis of optic flow patterns such as speed gradients and that the STS in humans plays a greater role in the analysis of 3D SFM than previously thought. PMID:19674088

Odontogenic Differentiation of Human Dental Pulp Stem Cells Stimulated by the Calcium Phosphate Porous Granules

PubMed Central

Nam, Sunyoung; Won, Jong-Eun; Kim, Cheol-Hwan; Kim, Hae-Won

2011-01-01

Effects of three-dimensional (3D) calcium phosphate (CaP) porous granules on the growth and odontogenic differentiation of human dental pulp stem cells (hDPSCs) were examined for dental tissue engineering. hDPSCs isolated from adult human dental pulps were cultured for 3-4 passages, and populated on porous granules. Cell growth on the culture dish showed an ongoing increase for up to 21 days, whereas the growth on the 3D granules decreased after 14 days. This reduction in proliferative potential on the 3D granules was more conspicuous under the osteogenic medium conditions, indicating that the 3D granules may induce the odontogenic differentiation of hDPSCs. Differentiation behavior on the 3D granules was confirmed by the increased alkaline phosphatase activity, up-regulation of odontoblast-specific genes, including dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) by quantitative polymerase chain reaction, and greater level of dentin sialoprotein synthesis by western blot. Moreover, the cellular mineralization, as assessed by Alizarin red S and calcium quantification, was significantly higher in the 3D CaP granules than in the culture dish. Taken all, the 3D CaP porous granules should be useful for dental tissue engineering in combination with hDPSCs by providing favorable 3D substrate conditions for cell growth and odontogenic development. PMID:21772958

Vitamin D fails to prevent serum starvation- or staurosporine-induced apoptosis in human and rat osteosarcoma-derived cell lines

DOE Office of Scientific and Technical Information (OSTI.GOV)

Witasp, Erika; Division of Biochemical Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm; Gustafsson, Ann-Catrin

2005-05-13

Previous studies have suggested that 1,25(OH){sub 2}D{sub 3}, the active form of vitamin D{sub 3}, may increase the survival of bone-forming osteoblasts through an inhibition of apoptosis. On the other hand, vitamin D{sub 3} has also been shown to trigger apoptosis in human cancer cells, including osteosarcoma-derived cell lines. In the present study, we show that 1,25(OH){sub 2}D{sub 3} induces a time- and dose-dependent loss of cell viability in the rat osteosarcoma cell line, UMR-106, and the human osteosarcoma cell line, TE-85. We were unable, however, to detect nuclear condensation, phosphatidylserine externalization, or other typical signs of apoptosis in thismore » model. Moreover, 1,25(OH){sub 2}D{sub 3} failed to protect against apoptosis induced by serum starvation or incubation with the protein kinase inhibitor, staurosporine. These in vitro findings are thus at variance with several previous reports in the literature and suggest that induction of or protection against apoptosis of bone-derived cells may not be a primary function of vitamin D{sub 3}.« less

Impact of Orientation on the Vitamin D Weighted Exposure of a Human in an Urban Environment

PubMed Central

Schrempf, Michael; Thuns, Nadine; Lange, Kezia

2017-01-01

The vitamin D3-weighted UV exposure of a human with vertical posture was calculated for urban locations to investigate the impact of orientation and obstructions on the exposure. Human exposure was calculated by using the 3D geometry of a human and integrating the radiance, i.e., the radiant energy from the direct solar beam and the diffuse sky radiation from different incident and azimuth angles. Obstructions of the sky are derived from hemispherical images, which are recorded by a digital camera with a fisheye lens. Due to the low reflectivity of most surfaces in the UV range, the radiance from obstructed sky regions was neglected. For spring equinox (21 March), the exposure of a human model with winter clothing in an environment where obstructions cover 40% of the sky varies by up to 25%, depending on the orientation of the human model to the sun. The calculation of the accumulated vitamin D3-weighted exposure of a human with winter clothing walking during lunch break shows that human exposure is reduced by the obstruction of buildings and vegetation by 40%. PMID:28813022

Development of a percentile based three-dimensional model of the buttocks in computer system

NASA Astrophysics Data System (ADS)

Wang, Lijing; He, Xueli; Li, Hongpeng

2016-05-01

There are diverse products related to human buttocks, which need to be designed, manufactured and evaluated with 3D buttock model. The 3D buttock model used in present research field is just simple approximate model similar to human buttocks. The 3D buttock percentile model is highly desired in the ergonomics design and evaluation for these products. So far, there is no research on the percentile sizing system of human 3D buttock model. So the purpose of this paper is to develop a new method for building three-dimensional buttock percentile model in computer system. After scanning the 3D shape of buttocks, the cloud data of 3D points is imported into the reverse engineering software (Geomagic) for the reconstructing of the buttock surface model. Five characteristic dimensions of the buttock are measured through mark-points after models being imported into engineering software CATIA. A series of space points are obtained by the intersecting of the cutting slices and 3D buttock surface model, and then are ordered based on the sequence number of the horizontal and vertical slices. The 1st, 5th, 50th, 95th, 99th percentile values of the five dimensions and the spatial coordinate values of the space points are obtained, and used to reconstruct percentile buttock models. This research proposes a establishing method of percentile sizing system of buttock 3D model based on the percentile values of the ischial tuberosities diameter, the distances from margin to ischial tuberosity and the space coordinates value of coordinate points, for establishing the Nth percentile 3D buttock model and every special buttock types model. The proposed method also serves as a useful guidance for the other 3D percentile models establishment for other part in human body with characteristic points.

A 3D human tissue-engineered lung model to study influenza A infection.

PubMed

Bhowmick, Rudra; Derakhshan, Mina; Liang, Yurong; Ritchey, Jerry; Liu, Lin; Gappa-Fahlenkamp, Heather

2018-05-05

Influenza A virus (IAV) claims approximately 250,000-500,000 lives annually worldwide. Currently, there are a few in vitro models available to study IAV immunopathology. Monolayer cultures of cell lines and primary lung cells (2D cell culture) is the most commonly used tool, however, this system does not have the in vivo-like structure of the lung and immune responses to IAV as it lacks the three-dimensional (3D) tissue structure. To recapitulate the lung physiology in vitro, a system that contains multiple cell types within a 3D environment that allows cell movement and interaction, would provide a critical tool. In this study, as a first step in designing a 3D-Human Tissue-Engineering Lung Model (3D-HTLM), we described the 3D culture of primary human small airway epithelial cells (HSAEpCs), and determined the immunophenotype of this system in response to IAV infections. We constructed a 3D chitosan-collagen scaffold and cultured HSAEpCs on these scaffolds at air-liquid interface (ALI). These 3D cultures were compared with 2D-cultured HSAEpCs for viability, morphology, marker protein expression, and cell differentiation. Results showed that the 3D-cultured HSAEpCs at ALI yielded maximum viable cells and morphologically resembled the in vivo lower airway epithelium. There were also significant increases in aquaporin-5 and cytokeratin-14 expression for HSAEpCs cultured in 3D compared to 2D. The 3D culture system was used to study the infection of HSAEpCs with two major IAV strains, H1N1 and H3N2.The HSAEpCs showed distinct changes in marker protein expression, both at mRNA and protein levels, and the release of proinflammatory cytokines. This study is the first step in the development of the 3D-HTLM, which will have wide applicability in studying pulmonary pathophysiology and therapeutics development.

Real-time multiple human perception with color-depth cameras on a mobile robot.

PubMed

Zhang, Hao; Reardon, Christopher; Parker, Lynne E

2013-10-01

The ability to perceive humans is an essential requirement for safe and efficient human-robot interaction. In real-world applications, the need for a robot to interact in real time with multiple humans in a dynamic, 3-D environment presents a significant challenge. The recent availability of commercial color-depth cameras allow for the creation of a system that makes use of the depth dimension, thus enabling a robot to observe its environment and perceive in the 3-D space. Here we present a system for 3-D multiple human perception in real time from a moving robot equipped with a color-depth camera and a consumer-grade computer. Our approach reduces computation time to achieve real-time performance through a unique combination of new ideas and established techniques. We remove the ground and ceiling planes from the 3-D point cloud input to separate candidate point clusters. We introduce the novel information concept, depth of interest, which we use to identify candidates for detection, and that avoids the computationally expensive scanning-window methods of other approaches. We utilize a cascade of detectors to distinguish humans from objects, in which we make intelligent reuse of intermediary features in successive detectors to improve computation. Because of the high computational cost of some methods, we represent our candidate tracking algorithm with a decision directed acyclic graph, which allows us to use the most computationally intense techniques only where necessary. We detail the successful implementation of our novel approach on a mobile robot and examine its performance in scenarios with real-world challenges, including occlusion, robot motion, nonupright humans, humans leaving and reentering the field of view (i.e., the reidentification challenge), human-object and human-human interaction. We conclude with the observation that the incorporation of the depth information, together with the use of modern techniques in new ways, we are able to create an accurate system for real-time 3-D perception of humans by a mobile robot.

Mapping of unknown industrial plant using ROS-based navigation mobile robot

NASA Astrophysics Data System (ADS)

Priyandoko, G.; Ming, T. Y.; Achmad, M. S. H.

2017-10-01

This research examines how humans work with teleoperated unmanned mobile robot inspection in industrial plant area resulting 2D/3D map for further critical evaluation. This experiment focuses on two parts, the way human-robot doing remote interactions using robust method and the way robot perceives the environment surround as a 2D/3D perspective map. ROS (robot operating system) as a tool was utilized in the development and implementation during the research which comes up with robust data communication method in the form of messages and topics. RGBD SLAM performs the visual mapping function to construct 2D/3D map using Kinect sensor. The results showed that the mobile robot-based teleoperated system are successful to extend human perspective in term of remote surveillance in large area of industrial plant. It was concluded that the proposed work is robust solution for large mapping within an unknown construction building.

Deterministically patterned biomimetic human iPSC-derived hepatic model via rapid 3D bioprinting

PubMed Central

Ma, Xuanyi; Qu, Xin; Zhu, Wei; Li, Yi-Shuan; Yuan, Suli; Zhang, Hong; Liu, Justin; Wang, Pengrui; Lai, Cheuk Sun Edwin; Zanella, Fabian; Feng, Gen-Sheng; Sheikh, Farah; Chien, Shu; Chen, Shaochen

2016-01-01

The functional maturation and preservation of hepatic cells derived from human induced pluripotent stem cells (hiPSCs) are essential to personalized in vitro drug screening and disease study. Major liver functions are tightly linked to the 3D assembly of hepatocytes, with the supporting cell types from both endodermal and mesodermal origins in a hexagonal lobule unit. Although there are many reports on functional 2D cell differentiation, few studies have demonstrated the in vitro maturation of hiPSC-derived hepatic progenitor cells (hiPSC-HPCs) in a 3D environment that depicts the physiologically relevant cell combination and microarchitecture. The application of rapid, digital 3D bioprinting to tissue engineering has allowed 3D patterning of multiple cell types in a predefined biomimetic manner. Here we present a 3D hydrogel-based triculture model that embeds hiPSC-HPCs with human umbilical vein endothelial cells and adipose-derived stem cells in a microscale hexagonal architecture. In comparison with 2D monolayer culture and a 3D HPC-only model, our 3D triculture model shows both phenotypic and functional enhancements in the hiPSC-HPCs over weeks of in vitro culture. Specifically, we find improved morphological organization, higher liver-specific gene expression levels, increased metabolic product secretion, and enhanced cytochrome P450 induction. The application of bioprinting technology in tissue engineering enables the development of a 3D biomimetic liver model that recapitulates the native liver module architecture and could be used for various applications such as early drug screening and disease modeling. PMID:26858399

Deterministically patterned biomimetic human iPSC-derived hepatic model via rapid 3D bioprinting.

PubMed

Ma, Xuanyi; Qu, Xin; Zhu, Wei; Li, Yi-Shuan; Yuan, Suli; Zhang, Hong; Liu, Justin; Wang, Pengrui; Lai, Cheuk Sun Edwin; Zanella, Fabian; Feng, Gen-Sheng; Sheikh, Farah; Chien, Shu; Chen, Shaochen

2016-02-23

The functional maturation and preservation of hepatic cells derived from human induced pluripotent stem cells (hiPSCs) are essential to personalized in vitro drug screening and disease study. Major liver functions are tightly linked to the 3D assembly of hepatocytes, with the supporting cell types from both endodermal and mesodermal origins in a hexagonal lobule unit. Although there are many reports on functional 2D cell differentiation, few studies have demonstrated the in vitro maturation of hiPSC-derived hepatic progenitor cells (hiPSC-HPCs) in a 3D environment that depicts the physiologically relevant cell combination and microarchitecture. The application of rapid, digital 3D bioprinting to tissue engineering has allowed 3D patterning of multiple cell types in a predefined biomimetic manner. Here we present a 3D hydrogel-based triculture model that embeds hiPSC-HPCs with human umbilical vein endothelial cells and adipose-derived stem cells in a microscale hexagonal architecture. In comparison with 2D monolayer culture and a 3D HPC-only model, our 3D triculture model shows both phenotypic and functional enhancements in the hiPSC-HPCs over weeks of in vitro culture. Specifically, we find improved morphological organization, higher liver-specific gene expression levels, increased metabolic product secretion, and enhanced cytochrome P450 induction. The application of bioprinting technology in tissue engineering enables the development of a 3D biomimetic liver model that recapitulates the native liver module architecture and could be used for various applications such as early drug screening and disease modeling.

Three-dimensional cell culture models for investigating human viruses.

PubMed

He, Bing; Chen, Guomin; Zeng, Yi

2016-10-01

Three-dimensional (3D) culture models are physiologically relevant, as they provide reproducible results, experimental flexibility and can be adapted for high-throughput experiments. Moreover, these models bridge the gap between traditional two-dimensional (2D) monolayer cultures and animal models. 3D culture systems have significantly advanced basic cell science and tissue engineering, especially in the fields of cell biology and physiology, stem cell research, regenerative medicine, cancer research, drug discovery, and gene and protein expression studies. In addition, 3D models can provide unique insight into bacteriology, virology, parasitology and host-pathogen interactions. This review summarizes and analyzes recent progress in human virological research with 3D cell culture models. We discuss viral growth, replication, proliferation, infection, virus-host interactions and antiviral drugs in 3D culture models.

Neurotrophin 3 upregulates proliferation and collagen production in human aortic valve interstitial cells: a potential role in aortic valve sclerosis.

PubMed

Yao, Qingzhou; Song, Rui; Ao, Lihua; Cleveland, Joseph C; Fullerton, David A; Meng, Xianzhong

2017-06-01

Calcific aortic valve disease (CAVD) is a leading cardiovascular disorder in the elderly. Diseased aortic valves are characterized by sclerosis (fibrosis) and nodular calcification. Sclerosis, an early pathological change, is caused by aortic valve interstitial cell (AVIC) proliferation and overproduction of extracellular matrix (ECM) proteins. However, the mechanism of aortic valve sclerosis remains unclear. Recently, we observed that diseased human aortic valves overexpress growth factor neurotrophin 3 (NT3). In the present study, we tested the hypothesis that NT3 is a profibrogenic factor to human AVICs. AVICs isolated from normal human aortic valves were cultured in M199 growth medium and treated with recombinant human NT3 (0.10 µg/ml). An exposure to NT3 induced AVIC proliferation, upregulated the production of collagen and matrix metalloproteinase (MMP), and augmented collagen deposition. These changes were abolished by inhibition of the Trk receptors. NT3 induced Akt phosphorylation and increased cyclin D1 protein levels in a Trk receptor-dependent fashion. Inhibition of Akt abrogated the effect of NT3 on cyclin D1 production. Furthermore, inhibition of either Akt or cyclin D1 suppressed NT3-induced cellular proliferation and MMP-9 and collagen production, as well as collagen deposition. Thus, NT3 upregulates cellular proliferation, ECM protein production, and collagen deposition in human AVICs. It exerts these effects through the Trk-Akt-cyclin D1 cascade. NT3 is a profibrogenic mediator in human aortic valve, and overproduction of NT3 by aortic valve tissue may contribute to the mechanism of valvular sclerosis. Copyright © 2017 the American Physiological Society.

3D abnormal behavior recognition in power generation

NASA Astrophysics Data System (ADS)

Wei, Zhenhua; Li, Xuesen; Su, Jie; Lin, Jie

2011-06-01

So far most research of human behavior recognition focus on simple individual behavior, such as wave, crouch, jump and bend. This paper will focus on abnormal behavior with objects carrying in power generation. Such as using mobile communication device in main control room, taking helmet off during working and lying down in high place. Taking account of the color and shape are fixed, we adopted edge detecting by color tracking to recognize object in worker. This paper introduces a method, which using geometric character of skeleton and its angle to express sequence of three-dimensional human behavior data. Then adopting Semi-join critical step Hidden Markov Model, weighing probability of critical steps' output to reduce the computational complexity. Training model for every behavior, mean while select some skeleton frames from 3D behavior sample to form a critical step set. This set is a bridge linking 2D observation behavior with 3D human joints feature. The 3D reconstruction is not required during the 2D behavior recognition phase. In the beginning of recognition progress, finding the best match for every frame of 2D observed sample in 3D skeleton set. After that, 2D observed skeleton frames sample will be identified as a specifically 3D behavior by behavior-classifier. The effectiveness of the proposed algorithm is demonstrated with experiments in similar power generation environment.

Implications of adipose-derived stromal cells in a 3D culture system for osteogenic differentiation: an in vitro and in vivo investigation.

PubMed

Shen, Francis H; Werner, Brian C; Liang, Haixiang; Shang, Hulan; Yang, Ning; Li, Xudong; Shimer, Adam L; Balian, Gary; Katz, Adam J

2013-01-01

Healthy mammalian cells in normal tissues are organized in complex three-dimensional (3D) networks that display nutrient and signaling gradients. Conventional techniques that grow cells in a 2D monolayer fail to reproduce the environment that is observed in vivo. In recent years, 3D culture systems have been used to mimic tumor microenvironments in cancer research and to emulate embryogenesis in stem cell cultures. However, there have been no studies exploring the ability for adipose-derived stromal (ADS) cells in a 3D culture system to undergo osteogenic differentiation. To characterize and investigate the in vitro and in vivo potential for human ADS cells in a novel 3D culture system to undergo osteogenic differentiation. Basic science and laboratory study. Human ADS cells were isolated and prepared as either a 2D monolayer or 3D multicellular aggregates (MAs). Multicellular aggregates were formed using the hanging droplet technique. Cells were treated in osteogenic medium in vitro, and cellular differentiation was investigated using gene expression, histology, and microCT at 1-, 2-, and 4-week time points. In vivo investigation involved creating a muscle pouch by developing the avascular muscular interval in the vastus lateralis of male athymic rats. Specimens were then pretreated with osteogenic medium and surgically implanted as (1) carrier (Matrigel) alone (control), (2) carrier with human ADS cells in monolayer, or (3) human ADS cells as MAs. In vivo evidence of osteogenic differentiation was evaluated with micro computed tomography and histologic sectioning at a 2-week time point. Human ADS cells cultured by the hanging droplet technique successfully formed MAs at the air-fluid interface. Adipose-derived stromal cells cultured in monolayer or as 3D MAs retain their ability to self-replicate and undergo multilineage differentiation as confirmed by increased runx2/Cbfa2, ALP, and OCN and increased matrix mineralization on histologic sectioning. Multicellular aggregate cells expressed increased differentiation potential and extracellular matrix production over the same human ADS cells cultured in monolayer. Furthermore, MAs reseeded onto monolayer retained their stem cell capabilities. When implanted in vivo, significantly greater bone volume and extracellular matrix were present in the implanted specimens of MAs confirmed on both microCT and histological sectioning. This is the first study to investigate the capability of human ADS cells in a 3D culture system to undergo osteogenic differentiation. The results confirm that MAs maintain their stem cell characteristics. Compared with analogous cells in monolayer culture, the human ADS cells as MAs exhibit elevated levels of osteogenic differentiation and increased matrix mineralization. Furthermore, the creation of uniform spheroids allows for improved handling and manipulation during transplantation. These findings strongly support the concept that 3D culture systems remain not only a viable option for stem cell culture but also possibly a more attractive alternative to traditional culture techniques to improve the osteogenic potential of human adipose stem cells. Copyright © 2013 Elsevier Inc. All rights reserved.

Elucidation of Chromatin Remodeling Machinery Involved in Regulation of Estrogen Receptor Alpha Expression in Human Breast Cancer Cells

DTIC Science & Technology

2006-08-01

depsipeptide with 5-aza-dC has been shown to synergistically reactivate silenced tumor suppressor genes in human cancer cells, including MLH1 , TIMP3...depsipeptide with 5- aza-dC has been shown to synergistically reactivate silenced tumor suppressor genes in human cancer cells, including MLH1 , TIMP3

Objective and subjective quality assessment of geometry compression of reconstructed 3D humans in a 3D virtual room

NASA Astrophysics Data System (ADS)

Mekuria, Rufael; Cesar, Pablo; Doumanis, Ioannis; Frisiello, Antonella

2015-09-01

Compression of 3D object based video is relevant for 3D Immersive applications. Nevertheless, the perceptual aspects of the degradation introduced by codecs for meshes and point clouds are not well understood. In this paper we evaluate the subjective and objective degradations introduced by such codecs in a state of art 3D immersive virtual room. In the 3D immersive virtual room, users are captured with multiple cameras, and their surfaces are reconstructed as photorealistic colored/textured 3D meshes or point clouds. To test the perceptual effect of compression and transmission, we render degraded versions with different frame rates in different contexts (near/far) in the scene. A quantitative subjective study with 16 users shows that negligible distortion of decoded surfaces compared to the original reconstructions can be achieved in the 3D virtual room. In addition, a qualitative task based analysis in a full prototype field trial shows increased presence, emotion, user and state recognition of the reconstructed 3D Human representation compared to animated computer avatars.

Vitamin D(3) synthesis in the entire skin surface of dairy cows despite hair coverage.

PubMed

Hymøller, L; Jensen, S K

2010-05-01

How hair-coated animals such as dairy cows synthesize endogenous vitamin D(3) during exposure to summer sunlight has been unclear since vitamin D(3) and its relation to sunlight was discovered. The fur of fur-bearing animals is thought to be comparable to clothing in humans, which prevents vitamin D(3) synthesis in the skin during exposure to sunlight. Different scenarios have been suggested but never tested in cows; for example, that vitamin D(3) is synthesized from sebum on the hair and ingested by cows during grooming or that body areas such as the udder and muzzle that have scant hair exclusively produce the vitamin. To test different scenarios, 16 Danish Holstein dairy cows were subjected to 4 degrees of coverage of their bodies with fabric that prevented vitamin D(3) synthesis in the covered skin areas. The treatments were horse blanket (cows fitted with horse blankets), udder cover (cows fitted with udder covers, horse blanket+udder cover (cows fitted with both horse blankets and udder covers), and natural (cows without any coverage fitted). The cows were let out to pasture daily between 1000 and 1500h for 4 wk in July and August 2009. Blood samples were collected 15 times during the study and analyzed for content of 25-hydroxyvitamin D(3) [25(OH)D(3)] indicative of the animals' vitamin D(3) status. Results showed that uncovered cows had a higher 25(OH)D(3) concentration in plasma after 28 d of access to sunlight compared with covered cows and that the plasma concentration of 25(OH)D(3) was strongly inversely correlated to the body surface area covered. These results are consistent with findings in humans, wherein the vitamin D(3) status of different individuals was inversely proportional to the amount of clothing worn during exposure to artificial sunlight. Hence, it appears that human clothing and cow hair are not comparable with respect to prevention of vitamin D(3) synthesis and that cows, like humans, synthesize vitamin D(3) evenly over their body surface. That vitamin D(3) should be synthesized from sebum on the hair and obtained by cows as a result of grooming is not supported by the findings in the present study either, because large differences were found between the treatment groups. If grooming were the source of vitamin D(3), then a relatively even 25(OH)D(3) concentration between treatments would be expected, because covered cows would obtain vitamin D(3) by grooming uncovered herdmates. Copyright 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

PET imaging predicts future body weight and cocaine preference

DOE Office of Scientific and Technical Information (OSTI.GOV)

Michaelides M.; Wang G.; Michaelides M.

Deficits in dopamine D2/D3 receptor (D2R/D3R) binding availability using PET imaging have been reported in obese humans and rodents. Similar deficits have been reported in cocaine-addicts and cocaine-exposed primates. We found that D2R/D3R binding availability negatively correlated with measures of body weight at the time of scan (ventral striatum), at 1 (ventral striatum) and 2 months (dorsal and ventral striatum) post scan in rats. Cocaine preference was negatively correlated with D2R/D3R binding availability 2 months (ventral striatum) post scan. Our findings suggest that inherent deficits in striatal D2R/D3R signaling are related to obesity and drug addiction susceptibility and that ventralmore » and dorsal striatum serve dissociable roles in maintaining weight gain and cocaine preference. Measuring D2R/D3R binding availability provides a way for assessing susceptibility to weight gain and cocaine abuse in rodents and given the translational nature of PET imaging, potentially primates and humans.« less

BINDING OF SOLUBLE IMMUNE COMPLEXES TO HUMAN LYMPHOBLASTOID CELLS

PubMed Central

Theofilopoulos, Argyrios N.; Dixon, Frank J.; Bokisch, Viktor A.

1974-01-01

In the present work we studied the expression of membrane-bound Ig (MBIg) as well as receptors for IgG Fc and complement on nine human lymphoblastoid cell lines. When MBIg and receptors for IgG Fc were compared, four categories of cell lines could be distinguished: (a) cell lines having both MBIg and receptors for IgG Fc, (b) cell lines having MBIg but lacking receptors for IgG Fc, (c) cell lines lacking MBIg but having receptors for IgG Fc, and (d) cell lines lacking both MBIg and receptors for IgG Fc. Two types of receptors for complement could be detected on the cell lines studied, one for C3-C3b and one for C3d. When sensitized red cells carrying C3b or C3d were used for rosette tests, three categories of cell lines could be distinguished: (a) cell lines having receptors for C3b and C3d, (b) cell lines having receptors only for C3d and (c) cell lines lacking both receptors. However, when a more sensitive immunofluorescent method was used instead of the rosette technique, it was found that cell lines unable to form rosettes with EAC1423bhu were able to bind soluble C3 or C3b which indicated the presence of these receptors on the cell surface. Inhibition experiments showed that receptors for C3-C3b and receptors for C3d are distinct and that receptors for C3-C3b and C3d are different from receptors for IgG Fc. A cell line (Raji) without MBIg but with receptors for IgG Fc, C3-C3b, and C3d was selected for use in studying the binding mechanism of soluble immune complexes to cell surface membrane. Aggregated human gamma globulin was used in place of immune complexes. Immune complexes containing complement bind to Raji cells only via receptors for complement, namely receptors for C3-C3b and C3d. Binding of immune complexes containing complement to cells is much greater than that of complexes without complement. Immune complexes bound to cells via receptors for complement can be partially released from the cell surface by addition of normal human serum as well as isolated human C3 or C3b. We postulate that such release is due to competition of immune complex bound C3b and free C3 or C3b for the receptors on Raji cells. PMID:4139225

3D Printed Stretchable Tactile Sensors.

PubMed

Guo, Shuang-Zhuang; Qiu, Kaiyan; Meng, Fanben; Park, Sung Hyun; McAlpine, Michael C

2017-07-01

The development of methods for the 3D printing of multifunctional devices could impact areas ranging from wearable electronics and energy harvesting devices to smart prosthetics and human-machine interfaces. Recently, the development of stretchable electronic devices has accelerated, concomitant with advances in functional materials and fabrication processes. In particular, novel strategies have been developed to enable the intimate biointegration of wearable electronic devices with human skin in ways that bypass the mechanical and thermal restrictions of traditional microfabrication technologies. Here, a multimaterial, multiscale, and multifunctional 3D printing approach is employed to fabricate 3D tactile sensors under ambient conditions conformally onto freeform surfaces. The customized sensor is demonstrated with the capabilities of detecting and differentiating human movements, including pulse monitoring and finger motions. The custom 3D printing of functional materials and devices opens new routes for the biointegration of various sensors in wearable electronics systems, and toward advanced bionic skin applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Improved Human Bone Marrow Mesenchymal Stem Cell Osteogenesis in 3D Bioprinted Tissue Scaffolds with Low Intensity Pulsed Ultrasound Stimulation

PubMed Central

Zhou, Xuan; Castro, Nathan J.; Zhu, Wei; Cui, Haitao; Aliabouzar, Mitra; Sarkar, Kausik; Zhang, Lijie Grace

2016-01-01

3D printing and ultrasound techniques are showing great promise in the evolution of human musculoskeletal tissue repair and regeneration medicine. The uniqueness of the present study was to combine low intensity pulsed ultrasound (LIPUS) and advanced 3D printing techniques to synergistically improve growth and osteogenic differentiation of human mesenchymal stem cells (MSC). Specifically, polyethylene glycol diacrylate bioinks containing cell adhesive Arginine-Glycine-Aspartic acid-Serene (RGDS) peptide and/or nanocrystalline hydroxyapatite (nHA) were used to fabricate 3D scaffolds with different geometric patterns via novel table-top stereolithography 3D printer. The resultant scaffolds provide a highly porous and interconnected 3D environment to support cell proliferation. Scaffolds with small square pores were determined to be the optimal geometric pattern for MSC attachment and growth. The optimal LIPUS working parameters were determined to be 1.5 MHz, 20% duty cycle with 150 mW/cm2 intensity. Results demonstrated that RGDS peptide and nHA containing 3D printed scaffolds under LIPUS treatment can greatly promote MSC proliferation, alkaline phosphatase activity, calcium deposition and total protein content. These results illustrate the effectiveness of the combination of LIPUS and biomimetic 3D printing scaffolds as a valuable combinatorial tool for improved MSC function, thus make them promising for future clinical and various regenerative medicine application. PMID:27597635

Simultaneous determination of TPN729 and its five metabolites in human plasma and urine by liquid chromatography coupled to tandem mass spectrometry.

PubMed

Liu, Yang; Shao, Shuguang; Song, Hanlin; Yao, Xueting; Liu, Jie; Liu, Hongzhong; Song, Ling; Jiang, Ji; Liu, Dongyang; Hu, Pei

2018-03-20

A specific and sensitive method was firstly developed using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) to simultaneously quantify TPN729 and its metabolites (TPN729-D1, TPN729-D2, TPN729M15-3 and TPN729M3) in human plasma and (TPN729-D1, TPN729-D2, TPN729M15-3 and TPN729M14) in human urine. Protein precipitation and direct dilution were used to extract TPN729 and its metabolites from plasma and urine, respectively. Ionization of TPN729, TPN729-D1, TPN729-D2, TPN729M15-3, TPN729M3, TPN729M14 and sildenafil (internal standard, IS) was performed using an electrospray ionization (ESI) source in positive mode and detection was carried out with multiple reaction monitoring (MRM) mode. This assay method for TPN729 and its five metabolites has been fully validated in terms of sensitivity, linearity, lower limit of quantification (LLOQ), precision, accuracy, stability, matrix effect and recovery. The LLOQ of TPN729/TPN729-D1/TPN729-D2/TPN729M15-3/TPN729M3 in human plasma and TPN729/TPN729-D1/TPN729-D2/TPN729M15-3/TPN729M14 in human urine were 0.200/0.500/2.00/0.500/1.00 ng/mL and 4.00/2.50/10.0/2.50/1.00 ng/mL, respectively. Inter- and intra-batch precision of TPN729 and its metabolites were less than 15% and the accuracy was within ±15% for both plasma and urine. The extraction recoveries of all analytes at three concentration levels were consistent. In conclusion, the validation results showed that this method was robust, specific, and sensitive and it can successfully fulfill the requirement of clinical pharmacokinetic study of TPN729 in Chinese healthy subjects. Copyright © 2018 Elsevier B.V. All rights reserved.

Analysis of discriminants for experimental 3D SAR imagery of human targets

NASA Astrophysics Data System (ADS)

Chan, Brigitte; Sévigny, Pascale; DiFilippo, David D. J.

2014-10-01

Development of a prototype 3-D through-wall synthetic aperture radar (SAR) system is currently underway at Defence Research and Development Canada. The intent is to map out building wall layouts and to detect targets of interest and their location behind walls such as humans, arms caches, and furniture. This situational awareness capability can be invaluable to the military working in an urban environment. Tools and algorithms are being developed to exploit the resulting 3-D imagery. Current work involves analyzing signatures of targets behind a wall and understanding the clutter and multipath signals in a room of interest. In this paper, a comprehensive study of 3-D human target signature metrics in free space is presented. The aim is to identify features for discrimination of the human target from other targets. Targets used in this investigation include a human standing, a human standing with arms stretched out, a chair, a table, and a metallic plate. Several features were investigated as potential discriminants and five which were identified as good candidates are presented in this paper. Based on this study, no single feature could be used to fully discriminate the human targets from all others. A combination of at least two different features is required to achieve this.

Human umbilical cord derived matrix: A scaffold suitable for tissue engineering application.

PubMed

Dan, Pan; Velot, Émilie; Mesure, Benjamin; Groshenry, Guillaume; Bacharouche, Jalal; Decot, Véronique; Menu, Patrick

2017-01-01

Human tissue derived natural extracellular matrix (ECM) has great potential in tissue engineering. We sought to isolate extracellular matrix derived from human umbilical cord and test its potential in tissue engineering. An enzymatic method was applied to isolate and solubilized complete human umbilical cord derived matrix (hUCM). The obtained solution was analyzed for growth factors, collagen and residual DNA contents, then used to coat 2D and 3D surfaces for cell culture application. The hUCM was successfully isolated with trypsin digestion to acquire a solution containing various growth factors and collagen but no residual DNA. This hUCM solution can form a coating on 2D and 3D substrates suitable cell culture. We developed a new matrix derived from human source that can be further used in tissue engineering.

Fast 3D NIR systems for facial measurement and lip-reading

NASA Astrophysics Data System (ADS)

Brahm, Anika; Ramm, Roland; Heist, Stefan; Rulff, Christian; Kühmstedt, Peter; Notni, Gunther

2017-05-01

Structured-light projection is a well-established optical method for the non-destructive contactless three-dimensional (3D) measurement of object surfaces. In particular, there is a great demand for accurate and fast 3D scans of human faces or facial regions of interest in medicine, safety, face modeling, games, virtual life, or entertainment. New developments of facial expression detection and machine lip-reading can be used for communication tasks, future machine control, or human-machine interactions. In such cases, 3D information may offer more detailed information than 2D images which can help to increase the power of current facial analysis algorithms. In this contribution, we present new 3D sensor technologies based on three different methods of near-infrared projection technologies in combination with a stereo vision setup of two cameras. We explain the optical principles of an NIR GOBO projector, an array projector and a modified multi-aperture projection method and compare their performance parameters to each other. Further, we show some experimental measurement results of applications where we realized fast, accurate, and irritation-free measurements of human faces.

Human Ozone (O3) Exposure Alters Serum Profile of Lipid Metabolites

EPA Science Inventory

HUMAN OZONE (O3) EXPOSURE ALTERS SERUM PROFILE OF LIPID METABOLITES Miller, D B.1; Kodavanti, U P.2 Karoly, E D.3; Cascio W.E2, Ghio, A J. 21. UNC-Chapel Hill, Chapel Hill, N.C., United States. 2. NHEERL, U.S. EPA, RTP, N.C., United States. 3. METABOLON INC., Durham, N.C., United...

3-D Model of the Human Respiratory System

EPA Science Inventory

The U.S. EPA’s Office of Research and Development (ORD) has developed a 3-D computational fluid dynamics (CFD) model of the human respiratory system that allows for the simulation of particulate based contaminant deposition and clearance, while being adaptable for age, ethnicity,...

A video, text, and speech-driven realistic 3-d virtual head for human-machine interface.

PubMed

Yu, Jun; Wang, Zeng-Fu

2015-05-01

A multiple inputs-driven realistic facial animation system based on 3-D virtual head for human-machine interface is proposed. The system can be driven independently by video, text, and speech, thus can interact with humans through diverse interfaces. The combination of parameterized model and muscular model is used to obtain a tradeoff between computational efficiency and high realism of 3-D facial animation. The online appearance model is used to track 3-D facial motion from video in the framework of particle filtering, and multiple measurements, i.e., pixel color value of input image and Gabor wavelet coefficient of illumination ratio image, are infused to reduce the influence of lighting and person dependence for the construction of online appearance model. The tri-phone model is used to reduce the computational consumption of visual co-articulation in speech synchronized viseme synthesis without sacrificing any performance. The objective and subjective experiments show that the system is suitable for human-machine interaction.

Evaluation of vision training using 3D play game

NASA Astrophysics Data System (ADS)

Kim, Jung-Ho; Kwon, Soon-Chul; Son, Kwang-Chul; Lee, Seung-Hyun

2015-03-01

The present study aimed to examine the effect of the vision training, which is a benefit of watching 3D video images (3D video shooting game in this study), focusing on its accommodative facility and vergence facility. Both facilities, which are the scales used to measure human visual performance, are very important factors for man in leading comfortable and easy life. This study was conducted on 30 participants in their 20s through 30s (19 males and 11 females at 24.53 ± 2.94 years), who can watch 3D video images and play 3D game. Their accommodative and vergence facility were measured before and after they watched 2D and 3D game. It turned out that their accommodative facility improved after they played both 2D and 3D games and more improved right after they played 3D game than 2D game. Likewise, their vergence facility was proved to improve after they played both 2D and 3D games and more improved soon after they played 3D game than 2D game. In addition, it was demonstrated that their accommodative facility improved to greater extent than their vergence facility. While studies have been so far conducted on the adverse effects of 3D contents, from the perspective of human factor, on the imbalance of visual accommodation and convergence, the present study is expected to broaden the applicable scope of 3D contents by utilizing the visual benefit of 3D contents for vision training.

Characterization of lipoproteins from the turtle, Trachemys scripta elegans, in fasted and fed states.

PubMed

Cain, William; Song, Li; Stephens, Gregory; Usher, David

2003-04-01

The lipid and apolipoprotein composition of VLDL, IDL, LDL, HDL(2) and HDL(3) were examined in the turtle, Trachemys scripta elegans, in fasted and fed states. The lipid composition of turtle lipoproteins was very similar to their human counterparts. The major apolipoprotein found in LDL, IDL and VLDL, which has a molecular weight of approximately 550 kD, is a homologue of apoB100. The major apolipoprotein found in both HDL(2) and HDL(3), has a molecular weight of 28-kD and is homologous to human apoA-I. HDL(3) also contains a 6.5 kD protein that is homologous to apoA-II, while HDL(2) has two low molecular weight proteins of 6 kD and 7 kD which are also found on the triglyceride rich lipoproteins (TRL). The 7 kD protein is homologous to apoC-III, while the 6 kD protein has a similar size and distribution as apoC-II or apoC-I. In addition, HDL(2) also possesses a protein of 15.8 kD that has no obvious mammalian homologue. In both size and apolipoprotein composition, turtle HDL(2) resembles human HDL(2b) while turtle HDL(3) resembles human HDL(3). In the fasted state, turtles contained very little TRL. When fed a high fat diet, the amount of IDL and LDL sized particles increased significantly.

Effects of supplemental vitamin D and calcium on oxidative DNA damage marker in normal colorectal mucosa: a randomized clinical trial.

PubMed

Fedirko, Veronika; Bostick, Roberd M; Long, Qi; Flanders, W Dana; McCullough, Marjorie L; Sidelnikov, Eduard; Daniel, Carrie R; Rutherford, Robin E; Shaukat, Aasma

2010-01-01

The exact antineoplastic effects of calcium and vitamin D(3) in the human colon are unclear. Animal and in vitro studies show that these two agents reduce oxidative stress; however, these findings have never been investigated in humans. To address this, we conducted a pilot, randomized, double-blind, placebo-controlled, 2 x 2 factorial clinical trial to test the effects of calcium and vitamin D(3) on a marker of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8-OH-dG), in the normal colorectal mucosa. Patients (N = 92) with at least one pathology-confirmed colorectal adenoma were treated with 2 g/d calcium and/or 800 IU/d vitamin D(3) versus placebo over 6 months. Overall labeling and colorectal crypt distribution of 8-OH-dG in biopsies of normal-appearing rectal mucosa were detected by standardized automated immunohistochemistry and quantified by image analysis. After 6 months of treatment, 8-OH-dG labeling along the full lengths of colorectal crypts decreased by 22% (P = 0.15) and 25% (P = 0.10) in the calcium and vitamin D(3) groups, respectively, but not in the calcium plus vitamin D(3) group. The estimated treatment effects were strongest among participants with higher baseline colon crypt vitamin D receptor expression (P = 0.05). Overall, these preliminary results indicate that calcium and vitamin D(3) may decrease oxidative DNA damage in the normal human colorectal mucosa, support the hypothesis that 8-OH-dG labeling in colorectal crypts is a treatable oxidative DNA damage biomarker of risk for colorectal neoplasms, and provide support for further investigation of calcium and vitamin D(3) as chemopreventive agents against colorectal neoplasms.

Vitamin D receptor-mediated control of Soggy, Wise, and Hairless gene expression in keratinocytes.

PubMed

Hsieh, Jui-Cheng; Estess, Rudolf C; Kaneko, Ichiro; Whitfield, G Kerr; Jurutka, Peter W; Haussler, Mark R

2014-02-01

The vitamin D receptor (VDR), but not its hormonal ligand, 1,25-dihydroxyvitamin D3 (1,25D), is required for the progression of the mammalian hair cycle. We studied three genes relevant to hair cycle signaling, DKKL1 (Soggy), SOSTDC1 (Wise), and HR (Hairless), to determine whether their expression is regulated by VDR and/or its 1,25D ligand. DKKL1 mRNA was repressed 49-72% by 1,25D in primary human and CCD-1106 KERTr keratinocytes; a functional vitamin D responsive element (VDRE) was identified at -9590 bp in murine Soggy. Similarly, SOSTDC1 mRNA was repressed 41-59% by 1,25D in KERTr and primary human keratinocytes; a functional VDRE was located at -6215 bp in human Wise. In contrast, HR mRNA was upregulated 1.56- to 2.77-fold by 1,25D in primary human and KERTr keratinocytes; a VDRE (TGGTGAgtgAGGACA) consisting of an imperfect direct repeat separated by three nucleotides (DR3) was identified at -7269 bp in the human Hairless gene that mediated dramatic induction, even in the absence of 1,25D ligand. In parallel, a DR4 thyroid hormone responsive element, TGGTGAggccAGGACA, was identified at +1304 bp in the human HR gene that conferred tri-iodothyronine (T3)-independent transcriptional activation. Because the thyroid hormone receptor controls HR expression in the CNS, whereas VDR functions in concert with the HR corepressor specifically in skin, a model is proposed wherein unliganded VDR upregulates the expression of HR, the gene product of which acts as a downstream comodulator to feedback-repress DKKL1 and SOSTDC1, resulting in integration of bone morphogenic protein and Wnt signaling to drive the mammalian hair cycle and/or influencing epidermal function.

Mir-30d suppresses cell proliferation of colon cancer cells by inhibiting cell autophagy and promoting cell apoptosis.

PubMed

Zhang, Rui; Xu, Jian; Zhao, Jian; Bai, Jinghui

2017-06-01

MiR-30 family plays an important role in the tumorigenesis of human cancers. The aim of the study is to investigate the role of miR-30d in human colon cancer cell lines and explore the molecular mechanism in the proliferation of colon cancer cells. The expression of miR-30d was determined by real-time polymerase chain reaction assay in colon cancer cell lines (HCT15, HCT116, HT-29, DLD-1, and SW480) and the results demonstrated that miR-30d level was significantly decreased in human colon cancer cell lines, compared with normal colon epithelial cell line. Transfection with miR-30d mimics inhibited cell proliferation, and transfection with miR-30d inhibitors significantly promoted cell viability of colon cancer cells. Furthermore, TargetScan analysis predicted that miR-30d interacted with messenger RNA on its 3' untranslated region of ATG5, phosphoinositide 3-kinase, and Beclin1 to negatively regulate cell autophagy in colon cancer cells. Moreover, transfection with miR-30d induced cell arrest at G2/M phase of HT-29 cells. Overexpression of miR-30d mimics inhibited cell viability probably due to the inhibition of cell autophagy and promotion of cell apoptosis. Thus, MiR-30d inhibited cell autophagy by directly targeting messenger RNA of ATG5, phosphoinositide 3-kinase, and Beclin1 and promoted cell apoptosis of human colon cancer cells. It is helpful to clarify the function of miR-30d in tumorigenesis of human cancers.

Intralaboratory and interlaboratory evaluation of the EpiDerm 3D human reconstructed skin micronucleus (RSMN) assay.

PubMed

Hu, Ting; Kaluzhny, Yulia; Mun, Greg C; Barnett, Brenda; Karetsky, Viktor; Wilt, Nathan; Klausner, Mitchell; Curren, Rodger D; Aardema, Marilyn J

2009-03-17

A novel in vitro human reconstructed skin micronucleus (RSMN) assay has been developed using the EpiDerm 3D human skin model [R. D. Curren, G. C. Mun, D. P. Gibson, and M. J. Aardema, Development of a method for assessing micronucleus induction in a 3D human skin model EpiDerm, Mutat. Res. 607 (2006) 192-204]. The RSMN assay has potential use in genotoxicity assessments as a replacement for in vivo genotoxicity assays that will be banned starting in 2009 according to the EU 7th Amendment to the Cosmetics Directive. Utilizing EpiDerm tissues reconstructed with cells from four different donors, intralaboratory and interlaboratory reproducibility of the RSMN assay were examined. Seven chemicals were evaluated in three laboratories using a standard protocol. Each chemical was evaluated in at least two laboratories and in EpiDerm tissues from at least two different donors. Three model genotoxins, mitomycin C (MMC), vinblastine sulfate (VB) and methyl methanesulfonate (MMS) induced significant, dose-related increases in cytotoxicity and MN induction in EpiDerm tissues. Conversely, four dermal non-carcinogens, 4-nitrophenol (4-NP), trichloroethylene (TCE), 2-ethyl-1,3-hexanediol (EHD), and 1,2-epoxydodecane (EDD) were negative in the RSMN assay. Results between tissues reconstructed from different donors were comparable. These results indicate the RSMN assay using the EpiDerm 3D human skin model is a promising new in vitro genotoxicity assay that allows evaluation of chromosome damage following "in vivo-like" dermal exposures.

Vitamin D and adipose tissue-more than storage.

PubMed

Mutt, Shivaprakash J; Hyppönen, Elina; Saarnio, Juha; Järvelin, Marjo-Riitta; Herzig, Karl-Heinz

2014-01-01

The pandemic increase in obesity is inversely associated with vitamin D levels. While a higher BMI was causally related to lower 25-hydroxyvitamin D (25(OH)D), no evidence was obtained for a BMI lowering effect by higher 25(OH)D. Some of the physiological functions of 1,25(OH)2D3 (1,25-dihydroxycholecalciferol or calcitriol) via its receptor within the adipose tissue have been investigated such as its effect on energy balance, adipogenesis, adipokine, and cytokine secretion. Adipose tissue inflammation has been recognized as the key component of metabolic disorders, e.g., in the metabolic syndrome. The adipose organ secretes more than 260 different proteins/peptides. However, the molecular basis of the interactions of 1,25(OH)2D3, vitamin D binding proteins (VDBPs) and nuclear vitamin D receptor (VDR) after sequestration in adipose tissue and their regulations are still unclear. 1,25(OH)2D3 and its inactive metabolites are known to inhibit the formation of adipocytes in mouse 3T3-L1 cell line. In humans, 1,25(OH)2D3 promotes preadipocyte differentiation under cell culture conditions. Further evidence of its important functions is given by VDR knock out (VDR(-/-)) and CYP27B1 knock out (CYP27B1 (-/-)) mouse models: Both VDR(-/-) and CYP27B1(-/-) models are highly resistant to the diet induced weight gain, while the specific overexpression of human VDR in adipose tissue leads to increased adipose tissue mass. The analysis of microarray datasets from human adipocytes treated with macrophage-secreted products up-regulated VDR and CYP27B1 genes indicating the capacity of adipocytes to even produce active 1,25(OH)2D3. Experimental studies demonstrate that 1,25(OH)2D3 has an active role in adipose tissue by modulating inflammation, adipogenesis and adipocyte secretion. Yet, further in vivo studies are needed to address the effects and the effective dosages of vitamin D in human adipose tissue and its relevance in the associated diseases.

Lysophospholipid presentation by CD1d and recognition by a human Natural Killer T-cell receptor

DOE Office of Scientific and Technical Information (OSTI.GOV)

López-Sagaseta, Jacinto; Sibener, Leah V.; Kung, Jennifer E.

2014-10-02

Invariant Natural Killer T (iNKT) cells use highly restricted {alpha}{beta} T cell receptors (TCRs) to probe the repertoire of lipids presented by CD1d molecules. Here, we describe our studies of lysophosphatidylcholine (LPC) presentation by human CD1d and its recognition by a native, LPC-specific iNKT TCR. Human CD1d presenting LPC adopts an altered conformation from that of CD1d presenting glycolipid antigens, with a shifted {alpha}1 helix resulting in an open A pocket. Binding of the iNKT TCR requires a 7-{angstrom} displacement of the LPC headgroup but stabilizes the CD1d-LPC complex in a closed conformation. The iNKT TCR CDR loop footprint onmore » CD1d-LPC is anchored by the conserved positioning of the CDR3{alpha} loop, whereas the remaining CDR loops are shifted, due in part to amino-acid differences in the CDR3{beta} and J{beta} segment used by this iNKT TCR. These findings provide insight into how lysophospholipids are presented by human CD1d molecules and how this complex is recognized by some, but not all, human iNKT cells.« less

Direct 3D cell-printing of human skin with functional transwell system.

PubMed

Kim, Byoung Soo; Lee, Jung-Seob; Gao, Ge; Cho, Dong-Woo

2017-06-06

Three-dimensional (3D) cell-printing has been emerging as a promising technology with which to build up human skin models by enabling rapid and versatile design. Despite the technological advances, challenges remain in the development of fully functional models that recapitulate complexities in the native tissue. Moreover, although several approaches have been explored for the development of biomimetic human skin models, the present skin models based on multistep fabrication methods using polydimethylsiloxane chips and commercial transwell inserts could be tackled by leveraging 3D cell-printing technology. In this paper, we present a new 3D cell-printing strategy for engineering a 3D human skin model with a functional transwell system in a single-step process. A hybrid 3D cell-printing system was developed, allowing for the use of extrusion and inkjet modules at the same time. We began by revealing the significance of each module in engineering human skin models; by using the extrusion-dispensing module, we engineered a collagen-based construct with polycaprolactone (PCL) mesh that prevented the contraction of collagen during tissue maturation; the inkjet-based dispensing module was used to uniformly distribute keratinocytes. Taking these features together, we engineered a human skin model with a functional transwell system; the transwell system and fibroblast-populated dermis were consecutively fabricated by using the extrusion modules. Following this process, keratinocytes were uniformly distributed onto the engineered dermis by the inkjet module. Our transwell system indicates a supportive 3D construct composed of PCL, enabling the maturation of a skin model without the aid of commercial transwell inserts. This skin model revealed favorable biological characteristics that included a stabilized fibroblast-stretched dermis and stratified epidermis layers after 14 days. It was also observed that a 50 times reduction in cost was achieved and 10 times less medium was used than in a conventional culture. Collectively, because this single-step process opens up chances for versatile designs, we envision that our cell-printing strategy could provide an attractive platform in engineering various human skin models.

CYP3A5 Mediates Effects of Cocaine on Human Neocorticogenesis: Studies using an In Vitro 3D Self-Organized hPSC Model with a Single Cortex-Like Unit.

PubMed

Lee, Chun-Ting; Chen, Jia; Kindberg, Abigail A; Bendriem, Raphael M; Spivak, Charles E; Williams, Melanie P; Richie, Christopher T; Handreck, Annelie; Mallon, Barbara S; Lupica, Carl R; Lin, Da-Ting; Harvey, Brandon K; Mash, Deborah C; Freed, William J

2017-02-01

Because of unavoidable confounding variables in the direct study of human subjects, it has been difficult to unravel the effects of prenatal cocaine exposure on the human fetal brain, as well as the cellular and biochemical mechanisms involved. Here, we propose a novel approach using a human pluripotent stem cell (hPSC)-based 3D neocortical organoid model. This model retains essential features of human neocortical development by encompassing a single self-organized neocortical structure, without including an animal-derived gelatinous matrix. We reported previously that prenatal cocaine exposure to rats during the most active period of neural progenitor proliferation induces cytoarchitectural changes in the embryonic neocortex. We also identified a role of CYP450 and consequent oxidative ER stress signaling in these effects. However, because of differences between humans and rodents in neocorticogenesis and brain CYP metabolism, translation of the research findings from the rodent model to human brain development is uncertain. Using hPSC 3D neocortical organoids, we demonstrate that the effects of cocaine are mediated through CYP3A5-induced generation of reactive oxygen species, inhibition of neocortical progenitor cell proliferation, induction of premature neuronal differentiation, and interruption of neural tissue development. Furthermore, knockdown of CYP3A5 reversed these cocaine-induced pathological phenotypes, suggesting CYP3A5 as a therapeutic target to mitigate the deleterious neurodevelopmental effects of prenatal cocaine exposure in humans. Moreover, 3D organoid methodology provides an innovative platform for identifying adverse effects of abused psychostimulants and pharmaceutical agents, and can be adapted for use in neurodevelopmental disorders with genetic etiologies.

Top3-Rmi1 dissolve Rad51-mediated D loops by a topoisomerase-based mechanism.

PubMed

Fasching, Clare L; Cejka, Petr; Kowalczykowski, Stephen C; Heyer, Wolf-Dietrich

2015-02-19

The displacement loop (D loop) is a DNA strand invasion product formed during homologous recombination. Disruption of nascent D loops prevents recombination, and during synthesis-dependent strand annealing (SDSA), disruption of D loops extended by DNA polymerase ensures a non-crossover outcome. The proteins implicated in D loop disruption are DNA motor proteins/helicases that act by moving DNA junctions. Here we report that D loops can also be disrupted by DNA topoisomerase 3 (Top3), and this disruption depends on Top3's catalytic activity. Yeast Top3 specifically disrupts D loops mediated by yeast Rad51/Rad54; protein-free D loops or D loop mediated by bacterial RecA protein or human RAD51/RAD54 resist dissolution. Also, the human Topoisomerase IIIa-RMI1-RMI2 complex is capable of dissolving D loops. Consistent with genetic data, we suggest that the extreme growth defect and hyper-recombination phenotype of Top3-deficient yeast cells is partially a result of unprocessed D loops. Copyright © 2015 Elsevier Inc. All rights reserved.

3D enamel thickness in Neandertal and modern human permanent canines.

PubMed

Buti, Laura; Le Cabec, Adeline; Panetta, Daniele; Tripodi, Maria; Salvadori, Piero A; Hublin, Jean-Jacques; Feeney, Robin N M; Benazzi, Stefano

2017-12-01

Enamel thickness figures prominently in studies of human evolution, particularly for taxonomy, phylogeny, and paleodietary reconstruction. Attention has focused on molar teeth, through the use of advanced imaging technologies and novel protocols. Despite the important results achieved thus far, further work is needed to investigate all tooth classes. We apply a recent approach developed for anterior teeth to investigate the 3D enamel thickness of Neandertal and modern human (MH) canines. In terms of crown size, the values obtained for both upper and lower unworn/slightly worn canines are significantly greater in Neandertals than in Upper Paleolithic and recent MH. The 3D relative enamel thickness (RET) is significantly lower in Neandertals than in MH. Moreover, differences in 3D RET values between the two groups appear to decrease in worn canines beginning from wear stage 3, suggesting that both the pattern and the stage of wear may have important effects on the 3D RET value. Nevertheless, the 3D average enamel thickness (AET) does not differ between the two groups. In both groups, 3D AET and 3D RET indices are greater in upper canines than in lower canines, and overall the enamel is thicker on the occlusal half of the labial aspect of the crown, particularly in MH. By contrast, the few early modern humans investigated show the highest volumes of enamel while for all other components of 3D enamel, thickness this group holds an intermediate position between Neandertals and recent MH. Overall, our study supports the general findings that Neandertals have relatively thinner enamel than MH (as also observed in molars), indicating that unworn/slightly worn canines can be successfully used to discriminate between the two groups. Further studies, however, are needed to understand whether these differences are functionally related or are the result of pleiotropic or genetic drift effects. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors.

PubMed

Li, Ling-Wei; Fan, Li-Qing; Zhu, Wen-Bing; Nien, Hong-Chuan; Sun, Bo-Lan; Luo, Ke-Li; Liao, Ting-Ting; Tang, Le; Lu, Guang-Xiu

2007-05-01

To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis (2-DE) reference map of human spermatozoal proteins in a pH range of 3.5-9.0. In order to reveal more protein spots, immobilized pH gradient strips (24 cm) of broad range of pH 3-10 and the narrower range of pH 6-9, as well as different overlapping narrow range pH immobilized pH gradient (IPG) strips, including 3.5-4.5, 4.0-5.0, 4.5-5.5, 5.0-6.0 and 5.5-6.7, were used. After 2-DE, several visually identical spots between the different pH range 2-D gel pairs were cut from the gels and confirmed by mass spectrometry and used as landmarks for computer analysis. The 2-D reference map with pH value from 3.5 to 9.0 was synthesized by using the ImageMaster analysis software. The overlapping spots were excluded, so that every spot was counted only once. A total of 3872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip (1306 spots). The present 2-D pattern is a high resolution 2-D reference map for human fertile spermatozoal protein spots. A comprehensive knowledge of the protein composition of human spermatozoa is very meaningful in studying dysregulation of male fertility.

Placental vitamin D metabolism and its associations with circulating vitamin D metabolites in pregnant women.

PubMed

Park, Heyjun; Wood, Madeleine R; Malysheva, Olga V; Jones, Sara; Mehta, Saurabh; Brannon, Patsy M; Caudill, Marie A

2017-12-01

Background: Little is known about placental vitamin D metabolism and its impact on maternal circulating vitamin D concentrations in humans. Objective: This study sought to advance the current understanding of placental vitamin D metabolism and its role in modulating maternal circulating vitamin D metabolites during pregnancy. Design: Nested within a feeding study, 24 healthy pregnant women (26-29 wk of gestation) consumed a single amount of vitamin D (511 IU/d from diet and a cholecalciferol supplement) for 10 wk. Concentrations of placental and blood vitamin D metabolites and placental messenger RNA (mRNA) abundance of vitamin D metabolic pathway components were quantified. In addition, cultured human trophoblasts were incubated with 13 C-cholecalciferol to examine the intracellular generation and secretion of vitamin D metabolites along with the regulation of target genes. Results: In placental tissue, 25-hydroxyvitamin D 3 [25(OH)D 3 ] was strongly correlated ( r = 0.83, P < 0.001) with 24,25-dihydroxyvitamin D 3 Moreover, these placental metabolites were strongly correlated ( r ≤ 0.85, P ≤ 0.04) with their respective metabolites in maternal circulation. Positive associations ( P ≤ 0.045) were also observed between placental mRNA abundance of vitamin D metabolic components and circulating vitamin D metabolites [i.e., LDL-related protein 2 ( LRP2 , also known as megalin) with 25(OH)D 3 and the C3 epimer of 25(OH)D 3 [3-epi-25(OH)D 3 ]; cubilin ( CUBN ) with 25(OH)D 3 ; 25-hydroxylase ( CYP2R1 ) with 3-epi-25(OH)D 3 ; 24-hydroxylase ( CYP24A1 ) with 25(OH)D 3 , 3-epi-25(OH)D 3 , and 1,25-dihydroxyvitamin D 3 [1,25(OH) 2 D 3 ]; and 1α-hydroxylase [( CYP27B1 ) with 3-epi-25(OH)D 3 and 1,25(OH) 2 D 3 ]. Notably, in vitro experiments with trophoblasts showed increased production and secretion of 25(OH)D 3 and higher CYP24A1 gene transcript abundance in response to cholecalciferol treatment. Conclusions: The numerous associations of many of the placental biomarkers of vitamin D metabolism with circulating vitamin D metabolites among pregnant women [including a CYP27B1 -associated increase in 1,25(OH) 2 D 3 ] and the evidence of trophoblast production and secretion of vitamin D metabolites, especially 25(OH)D 3 , suggest that the placenta may play an active role in modulating the vitamin D metabolite profile in maternal circulation in human pregnancy. This trial was registered at clinicaltrials.gov as NCT03051867. © 2017 American Society for Nutrition.

The Human Side of Cyber Conflict: Organizing, Training, and Equipping the Air Force Cyber Workforce

DTIC Science & Technology

2016-06-01

Breakdown of the 17D community as of 31 March 2014. (Reproduced from 17D Officer Assignment Team, Cyberspace Operations “Spread the Word” briefing, 9– 11 ...surety 3D0X4 Computer systems programs 3D1X1 Client systems 3D1X2 Cyber transport 3D1X3 Radio frequency transport 3D1X4 Spectrum operations 3D1X5 Radar...Computer systems programs 3D1X1 Client systems 3D1X2 Cyber transport systems 3D1X3 Radio frequency transmissionsystems FORCE DEVELOPMENT │ 123 Table 8

Development and Optimization of Viable Human Platforms through 3D Printing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parker, Paul R.; Moya, Monica L.; Wheeler, Elizabeth K.

2015-08-21

3D printing technology offers a unique method for creating cell cultures in a manner far more conducive to accurate representation of human tissues and systems. Here we print cellular structures capable of forming vascular networks and exhibiting qualities of natural tissues and human systems. This allows for cheaper and readily available sources for further study of biological and pharmaceutical agents.

The effectiveness and user perception of 3-dimensional digital human anatomy in an online undergraduate anatomy laboratory

NASA Astrophysics Data System (ADS)

Hilbelink, Amy Joanne

2007-12-01

The primary purpose of this study was to determine the effectiveness of implementing desktop 3-dimensional (3D) stereo images of human anatomy into an undergraduate human anatomy distance laboratory. User perceptions of 2D and 3D images were gathered via questionnaire in order to determine ease of use and level of satisfaction associated with the 3D software in the online learning environment. Mayer's (2001, p. 184) principles of design were used to develop the study materials that consisted of PowerPoint presentations and AVI files accessed via Blackboard. The research design employed a mixed-methods approach. Volunteers each were administered a demographic survey and were then stratified into groups based upon pre-test scores. A total sample size of 62 pairs was available for combined data analysis. Quantitative research questions regarding the effectiveness of 2D versus the 3D treatment were analyzed using a doubly-multivariate repeated measures (Doubly-MANOVA) design. Paired test scores achieved by undergraduates on a laboratory practical of identification and spatial relationships of the bones and features of a human skull were used in the analysis. The questionnaire designed to gather user perceptions consisted of quantitative and qualitative questions. Response frequencies were analyzed for the two groups and common themes were noted. Results revealed a statistically significant difference in group means for the main effect of the treatment groups 2D and 3D and for the variables of identification and relationship with the 3D group outperforming the 2D group on both dependent variables. Effect sizes were determined to be small, 0.215 for the identification variable and 0.359 for the relationship variable. Overall, all students liked the convenience of using PowerPoint and AVI files online. The 3D group felt their PowerPoint was more realistic than did the 2D group and both groups appreciated the detailed labeling of the online images. One third of the volunteers in the 3D group indicated that "eye strain" was what they liked least about working with the 3D images. Results indicate that desktop, stereo imaging may be incorporated effectively into online anatomy and physiology courses, but that more work needs to be done to ensure less eye strain.

Recent developments in stereoscopic and holographic 3D display technologies

NASA Astrophysics Data System (ADS)

Sarma, Kalluri

2014-06-01

Currently, there is increasing interest in the development of high performance 3D display technologies to support a variety of applications including medical imaging, scientific visualization, gaming, education, entertainment, air traffic control and remote operations in 3D environments. In this paper we will review the attributes of the various 3D display technologies including stereoscopic and holographic 3D, human factors issues of stereoscopic 3D, the challenges in realizing Holographic 3D displays and the recent progress in these technologies.

An efficient synthesis of 1α,25-dihydroxyvitamin D3 LC-biotin.

PubMed

Kattner, Lars; Bernardi, Dan

2017-10-01

In recent years the apparent impact of vitamin D deficiency on human health has gained increased awareness. Consequently, the development of appropriate assays to measure the status of medicinally most relevant vitamin D metabolites in human blood, serum or relevant tissue is continuously being improved. Particularly, assaying of 1α,25-dihydroxyvitamin D 3 , in turn considered as the most active metabolite, is mainly indicated in disorders leading to calcaemia or those resulting from an impaired 1α-hydroxylation of 25-hydroxyvitamin D 3 . Thus, in some competitive protein binding and ELISA assays, biotin-linked 1α,25-dihydroxyvitamin D 3 (1α,25-dihydroxyvitamin D 3 LC-biotin) is employed for measurement of actual calicitriol concentration. A new efficient synthesis of 1α,25-dihydroxyvitamin D 3 LC-biotin is described, starting with readily available vitamin D 2 , and combining a classical approach to access 1α,25-dihydroxyvitamin D 3 , appropriate OH-protective group transformations, and a C-3-O-alkylation, suitable to connect the biotin-linker in a reliable, selective and high yielding strategy. The developed methodology is applicable to the synthesis of a wide variety of C-3-OH-linked vitamin D 3 and D 2 derivatives. Copyright © 2017 Elsevier Ltd. All rights reserved.

EphA2 proteomics in human keratinocytes reveals a novel association with afadin and epidermal tight junctions.

PubMed

Perez White, Bethany E; Ventrella, Rosa; Kaplan, Nihal; Cable, Calvin J; Thomas, Paul M; Getsios, Spiro

2017-01-01

EphA2 is a receptor tyrosine kinase that helps to maintain epidermal tissue homeostasis. A proximity-dependent biotin identification (BioID) approach was used to identify proteins in close proximity to EphA2 within primary human keratinocytes and three-dimensional (3D) reconstituted human epidermis (RHE) cultures to map a putative protein interaction network for this membrane receptor that exhibits a polarized distribution in stratified epithelia. Although a subset of known EphA2 interactors were identified in the BioID screen, >97% were uniquely detected in keratinocytes with over 50% of these vicinal proteins only present in 3D human epidermal culture. Afadin (AFDN), a cytoskeletal and junction-associated protein, was present in 2D and 3D keratinocyte cultures, and validated as a so-far-unknown EphA2-interacting protein. Loss of EphA2 protein disrupted the subcellular distribution of afadin and occludin in differentiated keratinocytes, leading to impairment of tight junctions. Collectively, these studies illustrate the use of the BioID approach in order to map receptor interaction networks in 3D human epithelial cultures, and reveal a positive regulatory role for EphA2 in the organization of afadin and epidermal tight junctions. © 2017. Published by The Company of Biologists Ltd.

EphA2 proteomics in human keratinocytes reveals a novel association with afadin and epidermal tight junctions

PubMed Central

Perez White, Bethany E.; Ventrella, Rosa; Kaplan, Nihal; Cable, Calvin J.; Thomas, Paul M.

2017-01-01

ABSTRACT EphA2 is a receptor tyrosine kinase that helps to maintain epidermal tissue homeostasis. A proximity-dependent biotin identification (BioID) approach was used to identify proteins in close proximity to EphA2 within primary human keratinocytes and three-dimensional (3D) reconstituted human epidermis (RHE) cultures to map a putative protein interaction network for this membrane receptor that exhibits a polarized distribution in stratified epithelia. Although a subset of known EphA2 interactors were identified in the BioID screen, >97% were uniquely detected in keratinocytes with over 50% of these vicinal proteins only present in 3D human epidermal culture. Afadin (AFDN), a cytoskeletal and junction-associated protein, was present in 2D and 3D keratinocyte cultures, and validated as a so-far-unknown EphA2-interacting protein. Loss of EphA2 protein disrupted the subcellular distribution of afadin and occludin in differentiated keratinocytes, leading to impairment of tight junctions. Collectively, these studies illustrate the use of the BioID approach in order to map receptor interaction networks in 3D human epithelial cultures, and reveal a positive regulatory role for EphA2 in the organization of afadin and epidermal tight junctions. PMID:27815408

Regio- and Stereo-Selective Oxidation of a Cardiovascular Drug, Metoprolol, Mediated by Cytochrome P450 2D and 3A Enzymes in Marmoset Livers.

PubMed

Uehara, Shotaro; Ishii, Sakura; Uno, Yasuhiro; Inoue, Takashi; Sasaki, Erika; Yamazaki, Hiroshi

2017-08-01

A β -blocker, metoprolol, is one of the in vivo probes for human cytochrome P450 (P450) 2D6. Investigation of nonhuman primate P450 enzymes helps to improve the accuracy of the extrapolation of pharmacokinetic data from animals into humans. Common marmosets ( Callithrix jacchus ) are a potential primate model for preclinical research, but the detailed roles of marmoset P450 enzymes in metoprolol oxidation remain unknown. In this study, regio- and stereo-selectivity of metoprolol oxidations by a variety of P450 enzymes in marmoset and human livers were investigated in vitro. Although liver microsomes from cynomolgus monkeys and rats preferentially mediated S -metoprolol O -demethylation and R -metoprolol α -hydroxylation, respectively, those from humans, marmosets, minipigs, and dogs preferentially mediated R -metoprolol O -demethylation, in contrast to the slow rates of R - and S -metoprolol oxidation in mouse liver microsomes. R - and S -metoprolol O -demethylation activities in marmoset livers were strongly inhibited by quinidine and ketoconazole, and were significantly correlated with bufuralol 1'-hydroxylation and midazolam 1'-hydroxylation activities and also with P450 2D and 3A4 contents, which is different from the case in human livers that did not have any correlations with P450 3A-mediated midazolam 1'-hydroxylation. Recombinant human P450 2D6 enzyme and marmoset P450 2D6/3A4 enzymes effectively catalyzed R -metoprolol O -demethylation, comparable to the activities of human and marmoset liver microsomes, respectively. These results indicated that the major roles of P450 2D enzymes for the regio- and stereo-selectivity of metoprolol oxidation were similar between human and marmoset livers, but the minor roles of P450 3A enzymes were unique to marmosets. Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.

Labeling and Other Effects of Actinomycin D on Human Chromosomes*

PubMed Central

Miles, Charles P.

1970-01-01

3H-actinomycin D, a guanine-binding agent, labels fixed human chromosomes nonrandomly. Actinomycin D added in G2 inhibits secondary constrictions and breaks chromosomes. There is some tendency for label to be concentrated at the ends of chromosomes and near the centromere. Labeling with 3H-thymidine in the late stage of DNA synthesis shows a different pattern and in general lacks the telomeric concentrations. The sites of actinomycin D-induced breaks do not show good correspondence with the sites of actinomycin D label. Images PMID:5267140

Maintenance of human adipose derived stem cell (hASC) differentiation capabilities using a 3D culture.

PubMed

Lin, Ching-Yu; Huang, Chi-Hui; Wu, Yuan-Kun; Cheng, Nai-Chen; Yu, Jiashing

2014-07-01

In this study, 3D culture system for human adipose-derived stem cell (hASC) using a BioLevitator as the bioreactor for microcarrier-based cultures was established. During the culturing period, hASCs preferred to grow in crevices between microcarriers and a high viability was maintained even when reaching confluency. Adipogenic or osteogenic differential medium was used to induce hASCs and differential potentials of these cells were compared between 2D and 3D environments via RT-PCR and staining quantifications. CEBP/α gene expression was significant higher in 3D condition at day 21 (P < 0.05). Staining quantification indicates that cells cultured in 3D condition have significant better differentiation potential from day 14 to 21 for both adipogenic and osteogenic lineages (P < 0.01).

To generate a finite element model of human thorax using the VCH dataset

NASA Astrophysics Data System (ADS)

Shi, Hui; Liu, Qian

2009-10-01

Purpose: To generate a three-dimensional (3D) finite element (FE) model of human thorax which may provide the basis of biomechanics simulation for the study of design effect and mechanism of safety belt when vehicle collision. Methods: Using manually or semi-manually segmented method, the interested area can be segmented from the VCH (Visible Chinese Human) dataset. The 3D surface model of thorax is visualized by using VTK (Visualization Toolkit) and further translated into (Stereo Lithography) STL format, which approximates the geometry of solid model by representing the boundaries with triangular facets. The data in STL format need to be normalized into NURBS surfaces and IGES format using software such as Geomagic Studio to provide archetype for reverse engineering. The 3D FE model was established using Ansys software. Results: The generated 3D FE model was an integrated thorax model which could reproduce human's complicated structure morphology including clavicle, ribs, spine and sternum. It was consisted of 1 044 179 elements in total. Conclusions: Compared with the previous thorax model, this FE model enhanced the authenticity and precision of results analysis obviously, which can provide a sound basis for analysis of human thorax biomechanical research. Furthermore, using the method above, we can also establish 3D FE models of some other organizes and tissues utilizing the VCH dataset.

Correlation between a 2D Channelized Hotelling Observer and Human Observers in a Low-contrast Detection Task with Multi-slice Reading in CT

PubMed Central

Yu, Lifeng; Chen, Baiyu; Kofler, James M.; Favazza, Christopher P.; Leng, Shuai; Kupinski, Matthew A.; McCollough, Cynthia H.

2017-01-01

Purpose Model observers have been successfully developed and used to assess the quality of static 2D CT images. However, radiologists typically read images by paging through multiple 2D slices (i.e. multi-slice reading). The purpose of this study was to correlate human and model observer performance in a low-contrast detection task performed using both 2D and multi-slice reading, and to determine if the 2D model observer still correlate well with human observer performance in multi-slice reading. Methods A phantom containing 18 low-contrast spheres (6 sizes × 3 contrast levels) was scanned on a 192-slice CT scanner at 5 dose levels (CTDIvol = 27, 13.5, 6.8, 3.4, and 1.7 mGy), each repeated 100 times. Images were reconstructed using both filtered-backprojection (FBP) and an iterative reconstruction (IR) method (ADMIRE, Siemens). A 3D volume of interest (VOI) around each sphere was extracted and placed side-by-side with a signal-absent VOI to create a 2-alternative forced choice (2AFC) trial. Sixteen 2AFC studies were generated, each with 100 trials, to evaluate the impact of radiation dose, lesion size and contrast, and reconstruction methods on object detection. In total, 1600 trials were presented to both model and human observers. Three medical physicists acted as human observers and were allowed to page through the 3D volumes to make a decision for each 2AFC trial. The human observer performance was compared with the performance of a multi-slice channelized Hotelling observer (CHO_MS), which integrates multi-slice image data, and with the performance of previously validated CHO, which operates on static 2D images (CHO_2D). For comparison, the same 16 2AFC studies were also performed in a 2D viewing mode by the human observers and compared with the multi-slice viewing performance and the two CHO models. Results Human observer performance was well correlated with the CHO_2D performance in the 2D viewing mode (Pearson product-moment correlation coefficient R=0.972, 95% confidence interval (CI): 0.919 to 0.990) and with the CHO_MS performance in the multi-slice viewing mode (R=0.952, 95% CI: 0.865 to 0.984). The CHO_2D performance, calculated from the 2D viewing mode, also had a strong correlation with human observer performance in the multi-slice viewing mode (R=0.957, 95% CI: 879 to 0.985). Human observer performance varied between the multi-slice and 2D modes. One reader performed better in the multi-slice mode (p=0.013); whereas the other two readers showed no significant difference between the two viewing modes (p=0.057 and p=0.38). Conclusions A 2D CHO model is highly correlated with human observer performance in detecting spherical low contrast objects in multi-slice viewing of CT images. This finding provides some evidence for the use of a simpler, 2D CHO to assess image quality in clinically relevant CT tasks where multi-slice viewing is used. PMID:28555878

Correlation between a 2D channelized Hotelling observer and human observers in a low-contrast detection task with multislice reading in CT.

PubMed

Yu, Lifeng; Chen, Baiyu; Kofler, James M; Favazza, Christopher P; Leng, Shuai; Kupinski, Matthew A; McCollough, Cynthia H

2017-08-01

Model observers have been successfully developed and used to assess the quality of static 2D CT images. However, radiologists typically read images by paging through multiple 2D slices (i.e., multislice reading). The purpose of this study was to correlate human and model observer performance in a low-contrast detection task performed using both 2D and multislice reading, and to determine if the 2D model observer still correlate well with human observer performance in multislice reading. A phantom containing 18 low-contrast spheres (6 sizes × 3 contrast levels) was scanned on a 192-slice CT scanner at five dose levels (CTDI vol = 27, 13.5, 6.8, 3.4, and 1.7 mGy), each repeated 100 times. Images were reconstructed using both filtered-backprojection (FBP) and an iterative reconstruction (IR) method (ADMIRE, Siemens). A 3D volume of interest (VOI) around each sphere was extracted and placed side-by-side with a signal-absent VOI to create a 2-alternative forced choice (2AFC) trial. Sixteen 2AFC studies were generated, each with 100 trials, to evaluate the impact of radiation dose, lesion size and contrast, and reconstruction methods on object detection. In total, 1600 trials were presented to both model and human observers. Three medical physicists acted as human observers and were allowed to page through the 3D volumes to make a decision for each 2AFC trial. The human observer performance was compared with the performance of a multislice channelized Hotelling observer (CHO_MS), which integrates multislice image data, and with the performance of previously validated CHO, which operates on static 2D images (CHO_2D). For comparison, the same 16 2AFC studies were also performed in a 2D viewing mode by the human observers and compared with the multislice viewing performance and the two CHO models. Human observer performance was well correlated with the CHO_2D performance in the 2D viewing mode [Pearson product-moment correlation coefficient R = 0.972, 95% confidence interval (CI): 0.919 to 0.990] and with the CHO_MS performance in the multislice viewing mode (R = 0.952, 95% CI: 0.865 to 0.984). The CHO_2D performance, calculated from the 2D viewing mode, also had a strong correlation with human observer performance in the multislice viewing mode (R = 0.957, 95% CI: 879 to 0.985). Human observer performance varied between the multislice and 2D modes. One reader performed better in the multislice mode (P = 0.013); whereas the other two readers showed no significant difference between the two viewing modes (P = 0.057 and P = 0.38). A 2D CHO model is highly correlated with human observer performance in detecting spherical low contrast objects in multislice viewing of CT images. This finding provides some evidence for the use of a simpler, 2D CHO to assess image quality in clinically relevant CT tasks where multislice viewing is used. © 2017 American Association of Physicists in Medicine.

Imaging of human differentiated 3D neural aggregates using light sheet fluorescence microscopy.

PubMed

Gualda, Emilio J; Simão, Daniel; Pinto, Catarina; Alves, Paula M; Brito, Catarina

2014-01-01

The development of three dimensional (3D) cell cultures represents a big step for the better understanding of cell behavior and disease in a more natural like environment, providing not only single but multiple cell type interactions in a complex 3D matrix, highly resembling physiological conditions. Light sheet fluorescence microscopy (LSFM) is becoming an excellent tool for fast imaging of such 3D biological structures. We demonstrate the potential of this technique for the imaging of human differentiated 3D neural aggregates in fixed and live samples, namely calcium imaging and cell death processes, showing the power of imaging modality compared with traditional microscopy. The combination of light sheet microscopy and 3D neural cultures will open the door to more challenging experiments involving drug testing at large scale as well as a better understanding of relevant biological processes in a more realistic environment.

3D printing of a wearable personalized oral delivery device: A first-in-human study

PubMed Central

Brambilla, Davide

2018-01-01

Despite the burgeoning interest in three-dimensional (3D) printing for the manufacture of customizable oral dosage formulations, a U.S. Food and Drug Administration–approved tablet notwithstanding, the full potential of 3D printing in pharmaceutical sciences has not been realized. In particular, 3D-printed drug-eluting devices offer the possibility for personalization in terms of shape, size, and architecture, but their clinical applications have remained relatively unexplored. We used 3D printing to manufacture a tailored oral drug delivery device with customizable design and tunable release rates in the form of a mouthguard and, subsequently, evaluated the performance of this system in the native setting in a first-in-human study. Our proof-of-concept work demonstrates the immense potential of 3D printing as a platform for the development and translation of next-generation drug delivery devices for personalized therapy. PMID:29750201

Features specific to retinal pigment epithelium cells derived from three-dimensional human embryonic stem cell cultures - a new donor for cell therapy.

PubMed

Wu, Wei; Zeng, Yuxiao; Li, Zhengya; Li, Qiyou; Xu, Haiwei; Yin, Zheng Qin

2016-04-19

Retinal pigment epithelium (RPE) transplantation is a particularly promising treatment of retinal degenerative diseases affecting RPE-photoreceptor complex. Embryonic stem cells (ESCs) provide an abundant donor source for RPE transplantation. Herein, we studied the time-course characteristics of RPE cells derived from three-dimensional human ESCs cultures (3D-RPE). We showed that 3D-RPE cells possessed morphology, ultrastructure, gene expression profile, and functions of authentic RPE. As differentiation proceeded, 3D-RPE cells could mature gradually with decreasing proliferation but increasing functions. Besides, 3D-RPE cells could form polarized monolayer with functional tight junction and gap junction. When grafted into the subretinal space of Royal College of Surgeons rats, 3D-RPE cells were safe and efficient to rescue retinal degeneration. This study showed that 3D-RPE cells were a new donor for cell therapy of retinal degenerative diseases.

Imaging of human differentiated 3D neural aggregates using light sheet fluorescence microscopy

PubMed Central

Gualda, Emilio J.; Simão, Daniel; Pinto, Catarina; Alves, Paula M.; Brito, Catarina

2014-01-01

The development of three dimensional (3D) cell cultures represents a big step for the better understanding of cell behavior and disease in a more natural like environment, providing not only single but multiple cell type interactions in a complex 3D matrix, highly resembling physiological conditions. Light sheet fluorescence microscopy (LSFM) is becoming an excellent tool for fast imaging of such 3D biological structures. We demonstrate the potential of this technique for the imaging of human differentiated 3D neural aggregates in fixed and live samples, namely calcium imaging and cell death processes, showing the power of imaging modality compared with traditional microscopy. The combination of light sheet microscopy and 3D neural cultures will open the door to more challenging experiments involving drug testing at large scale as well as a better understanding of relevant biological processes in a more realistic environment. PMID:25161607

Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics

PubMed Central

Vorrink, Sabine U.; Ullah, Shahid; Schmidt, Staffan; Nandania, Jatin; Velagapudi, Vidya; Beck, Olof; Ingelman-Sundberg, Magnus; Lauschke, Volker M.

2017-01-01

Adverse reactions or lack of response to medications are important concerns for drug development programs. However, faithful predictions of drug metabolism and toxicity are difficult because animal models show only limited translatability to humans. Furthermore, current in vitro systems, such as hepatic cell lines or primary human hepatocyte (PHH) 2-dimensional (2D) monolayer cultures, can be used only for acute toxicity tests because of their immature phenotypes and inherent instability. Therefore, the migration to novel phenotypically stable models is of prime importance for the pharmaceutical industry. Novel 3-dimensional (3D) culture systems have been shown to accurately mimic in vivo hepatic phenotypes on transcriptomic and proteomic level, but information about their metabolic stability is lacking. Using a combination of targeted and untargeted high-resolution mass spectrometry, we found that PHHs in 3D spheroid cultures remained metabolically stable for multiple weeks, whereas metabolic patterns of PHHs from the same donors cultured as conventional 2D monolayers rapidly deteriorated. Furthermore, pharmacokinetic differences between donors were maintained in 3D spheroid cultures, enabling studies of interindividual variability in drug metabolism and toxicity. We conclude that the 3D spheroid system is metabolically stable and constitutes a suitable model for in vitro studies of long-term drug metabolism and pharmacokinetics.—Vorrink, S. U., Ullah, S., Schmid, S., Nandania, J., Velagapudi, V., Beck, O., Ingelman-Sundberg, M., Lauschke, V. M. Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics. PMID:28264975

Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics.

PubMed

Vorrink, Sabine U; Ullah, Shahid; Schmidt, Staffan; Nandania, Jatin; Velagapudi, Vidya; Beck, Olof; Ingelman-Sundberg, Magnus; Lauschke, Volker M

2017-06-01

Adverse reactions or lack of response to medications are important concerns for drug development programs. However, faithful predictions of drug metabolism and toxicity are difficult because animal models show only limited translatability to humans. Furthermore, current in vitro systems, such as hepatic cell lines or primary human hepatocyte (PHH) 2-dimensional (2D) monolayer cultures, can be used only for acute toxicity tests because of their immature phenotypes and inherent instability. Therefore, the migration to novel phenotypically stable models is of prime importance for the pharmaceutical industry. Novel 3-dimensional (3D) culture systems have been shown to accurately mimic in vivo hepatic phenotypes on transcriptomic and proteomic level, but information about their metabolic stability is lacking. Using a combination of targeted and untargeted high-resolution mass spectrometry, we found that PHHs in 3D spheroid cultures remained metabolically stable for multiple weeks, whereas metabolic patterns of PHHs from the same donors cultured as conventional 2D monolayers rapidly deteriorated. Furthermore, pharmacokinetic differences between donors were maintained in 3D spheroid cultures, enabling studies of interindividual variability in drug metabolism and toxicity. We conclude that the 3D spheroid system is metabolically stable and constitutes a suitable model for in vitro studies of long-term drug metabolism and pharmacokinetics.-Vorrink, S. U., Ullah, S., Schmid, S., Nandania, J., Velagapudi, V., Beck, O., Ingelman-Sundberg, M., Lauschke, V. M. Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics. © The Author(s).

Exercise-induced TBC1D1 Ser237 phosphorylation and 14-3-3 protein binding capacity in human skeletal muscle.

PubMed

Frøsig, Christian; Pehmøller, Christian; Birk, Jesper B; Richter, Erik A; Wojtaszewski, Jørgen F P

2010-11-15

TBC1D1 is a Rab-GTPase activating protein involved in regulation of GLUT4 translocation in skeletal muscle. We here evaluated exercise-induced regulation of TBC1D1 Ser237 phosphorylation and 14-3-3 protein binding capacity in human skeletal muscle. In separate experiments healthy men performed all-out cycle exercise lasting either 30 s, 2  min or 20  min. After all exercise protocols, TBC1D1 Ser237 phosphorylation increased (∼70-230%, P < 0.005), with the greatest response observed after 20  min of cycling. Interestingly, capacity of TBC1D1 to bind 14-3-3 protein showed a similar pattern of regulation, increasing 60-250% (P < 0.001). Furthermore, recombinant 5AMP-activated protein kinase (AMPK) induced both Ser237 phosphorylation and 14-3-3 binding properties on human TBC1D1 when evaluated in vitro. To further characterize the role of AMPK as an upstream kinase regulating TBC1D1, extensor digitorum longus muscle (EDL) from whole body α1 or α2 AMPK knock-out and wild-type mice were stimulated to contract in vitro. In wild-type and α1 knock-out mice, contractions resulted in a similar ∼100% increase (P < 0.001) in Ser237 phosphorylation. Interestingly, muscle of α2 knock-out mice were characterized by reduced protein content of TBC1D1 (∼50%, P < 0.001) as well as in basal and contraction-stimulated (∼60%, P < 0.001) Ser237 phosphorylation, even after correction for the reduced TBC1D1 protein content. This study shows that TBC1D1 is Ser237 phosphorylated and 14-3-3 protein binding capacity is increased in response to exercise in human skeletal muscle. Furthermore, we show that the catalytic α2 AMPK subunit is the main (but probably not the only) donor of AMPK activity regulating TBC1D1 Ser237 phosphorylation in mouse EDL muscle.

Phenylbutyrate Is Bacteriostatic against Mycobacterium tuberculosis and Regulates the Macrophage Response to Infection, Synergistically with 25-Hydroxy-Vitamin D3.

PubMed

Coussens, Anna K; Wilkinson, Robert J; Martineau, Adrian R

2015-07-01

Adjunctive vitamin D treatment for pulmonary tuberculosis enhances resolution of inflammation but has modest effects on bacterial clearance. Sodium 4-phenylbutyrate (PBA) is in clinical use for a range of conditions and has been shown to synergise with vitamin D metabolites to upregulate cathelicidin antimicrobial peptide (CAMP) expression. We investigated whether clinically attainable plasma concentrations of PBA (0.4-4 mM) directly affect Mycobacterium tuberculosis (Mtb) growth and human macrophage and PBMC response to infection. We also tested the ability of PBA to enhance the immunomodulatory actions of the vitamin D metabolite 25(OH)D3 during infection and synergistically inhibit intracellular Mtb growth. PBA inhibited Mtb growth in broth with an MIC99 of 1 mM, which was reduced to 0.25 mM by lowering pH. During human macrophage infection, PBA treatment restricted Mtb uptake, phagocytic receptor expression and intracellular growth in a dose-dependent manner. PBA independently regulated CCL chemokine secretion and induced expression of the antimicrobial LTF (lactoferrin), the anti-inflammatory PROC (protein C) and multiple genes within the NLRP3 inflammasome pathway. PBA co-treatment with 25(OH)D3 synergistically modulated expression of numerous vitamin D-response genes, including CAMP, CYP24A1, CXCL10 and IL-37. This synergistic effect was dependent on MAPK signalling, while the effect of PBA on LTF, PROC and NLRP3 was MAPK-independent. During PBA and 25(OH)D3 co-treatment of human macrophages, in the absence of exogenous proteinase 3 (PR3) to activate cathelicidin, Mtb growth restriction was dominated by the effect of PBA, while the addition of PR3 enhanced growth restriction by 25(OH)D3 and PBA co-treatment. This suggests that PBA augments vitamin D-mediated cathelicidin-dependent Mtb growth restriction by human macrophages and independently induces antimicrobial and anti-inflammatory action. Therefore through both host-directed and bacterial-directed mechanisms PBA and vitamin D may prove an effective combinatorial adjunct therapy for tuberculosis to both resolve immunopathology and enhance bacterial clearance.

D Visibility Analysis in Urban Environment - Cognition Research Based on Vge

NASA Astrophysics Data System (ADS)

Lin, T. P.; Lin, H.; Hu, M. Y.

2013-09-01

The author in this research attempts to illustrate a measurable relationship between the physical environment and human's visual perception, including the distance, visual angle impact and visual field (a 3D isovist conception) against human's cognition way, by using a 3D visibility analysis method based on the platform of Virtual Geographic Environment (VGE). The whole project carries out in the CUHK campus (the Chinese University of Hong Kong), by adopting a virtual 3D model of the whole campus and survey in real world. A possible model for the simulation of human cognition in urban spaces is expected to be the output of this research, such as what the human perceive from the environment, how their feelings and behaviours are and how they affect the surrounding world. Kevin Lynch raised 5 elements of urban design in 1960s, which are "vitality, sense, fit, access and control". As the development of urban design, several problems around the human's cognitive and behaviour have come out. Due to the restriction of sensing knowledge in urban spaces, the research among the "sense" and the "fit" of urban design were not quite concerned in recent decades. The geo-spatial cognition field comes into being in 1997 and developed in recent 15 years, which made great effort in way-finding and urban behaviour simulation based on the platform of GIS (geographic information system) or VGE. The platform of VGE is recognized as a proper tool for the analysis of human's perception in urban places, because of its efficient 3D spatial data management and excellent 3D visualization for output result. This article will generally describe the visibility analysis method based on the 3D VGE platform. According to the uncertainty and variety of human perception existed in this research, the author attempts to arrange a survey of observer investigation and validation for the analysis results. Four figures related with space and human's perception will be mainly concerned in this proposal: openness, permeability, environmental pressure and visibility, and these will also be used as the identification for different type of spaces. Generally, the author is aiming at contributing a possible way to understand the reason of human's cognition in geo-spatial area, and provides efficient mathematical model between spatial information and visual perception to the related research field.

Adipogenesis of human adipose-derived stem cells within three-dimensional hollow fiber-based bioreactors.

PubMed

Gerlach, Jörg C; Lin, Yen-Chih; Brayfield, Candace A; Minteer, Danielle M; Li, Han; Rubin, J Peter; Marra, Kacey G

2012-01-01

To further differentiate adipose-derived stem cells (ASCs) into mature adipocytes and create three-dimensional (3D) adipose tissue in vitro, we applied multicompartment hollow fiber-based bioreactor technology with decentral mass exchange for more physiological substrate gradients and integral oxygenation. We hypothesize that a dynamic 3D perfusion in such a bioreactor will result in longer-term culture of human adipocytes in vitro, thus providing metabolically active tissue serving as a diagnostic model for screening drugs to treat diabetes. ASCs were isolated from discarded human abdominal subcutaneous adipose tissue and then inoculated into dynamic 3D culture bioreactors to undergo adipogenic differentiation. Insulin-stimulated glucose uptake from the medium was assessed with and without TNF-alpha. 3D adipose tissue was generated in the 3D-bioreactors. Immunohistochemical staining indicated that 3D-bioreactor culture displayed multiple mature adipocyte markers with more unilocular morphologies as compared with two-dimensional (2D) cultures. Results of real-time polymerase chain reaction showed 3D-bioreactor treatment had more efficient differentiation in fatty acid-binding protein 4 expression. Repeated insulin stimulation resulted in increased glucose uptake, with a return to baseline between testing. Importantly, TNF-alpha inhibited glucose uptake, an indication of the metabolic activity of the tissue. 3D bioreactors allow more mature adipocyte differentiation of ASCs compared with traditional 2D culture and generate adipose tissue in vitro for up to 2 months. Reproducible metabolic activity of the adipose tissue in the bioreactor was demonstrated, which is potentially useful for drug discovery. We present here, to the best of our knowledge for the first time, the development of a coherent 3D high density fat-like tissue consisting of unilocular structure from primary adipose stem cells in vitro.

Adipogenesis of Human Adipose-Derived Stem Cells Within Three-Dimensional Hollow Fiber-Based Bioreactors

PubMed Central

Gerlach, Jörg C.; Lin, Yen-Chih; Brayfield, Candace A.; Minteer, Danielle M.; Li, Han; Rubin, J. Peter

2012-01-01

To further differentiate adipose-derived stem cells (ASCs) into mature adipocytes and create three-dimensional (3D) adipose tissue in vitro, we applied multicompartment hollow fiber-based bioreactor technology with decentral mass exchange for more physiological substrate gradients and integral oxygenation. We hypothesize that a dynamic 3D perfusion in such a bioreactor will result in longer-term culture of human adipocytes in vitro, thus providing metabolically active tissue serving as a diagnostic model for screening drugs to treat diabetes. ASCs were isolated from discarded human abdominal subcutaneous adipose tissue and then inoculated into dynamic 3D culture bioreactors to undergo adipogenic differentiation. Insulin-stimulated glucose uptake from the medium was assessed with and without TNF-alpha. 3D adipose tissue was generated in the 3D-bioreactors. Immunohistochemical staining indicated that 3D-bioreactor culture displayed multiple mature adipocyte markers with more unilocular morphologies as compared with two-dimensional (2D) cultures. Results of real-time polymerase chain reaction showed 3D-bioreactor treatment had more efficient differentiation in fatty acid-binding protein 4 expression. Repeated insulin stimulation resulted in increased glucose uptake, with a return to baseline between testing. Importantly, TNF-alpha inhibited glucose uptake, an indication of the metabolic activity of the tissue. 3D bioreactors allow more mature adipocyte differentiation of ASCs compared with traditional 2D culture and generate adipose tissue in vitro for up to 2 months. Reproducible metabolic activity of the adipose tissue in the bioreactor was demonstrated, which is potentially useful for drug discovery. We present here, to the best of our knowledge for the first time, the development of a coherent 3D high density fat-like tissue consisting of unilocular structure from primary adipose stem cells in vitro. PMID:21902468

MART-10 represses cholangiocarcinoma cell growth and high vitamin D receptor expression indicates better prognosis for cholangiocarcinoma

PubMed Central

Chiang, Kun-Chun; Yeh, Ta-Sen; Huang, Cheng-Cheng; Chang, Yu-Chan; Juang, Horng-Heng; Cheng, Chi-Tung; Pang, Jong-Hwei S.; Hsu, Jun-Te; Takano, Masashi; Chen, Tai C.; Kittaka, Atsushi; Hsiao, Michael; Yeh, Chun-Nan

2017-01-01

Cholangiocarcinoma (CCA) is a devastating disease due to no effective treatments available. Since the non-mineral functions of vitamin D emerges, 1α,25(OH)2D3, the active form of vitamin D, has been applied in anti-cancer researches. In this study, we demonstrated that both the 1α,25(OH)2D3 analog, MART-10, and 1α,25(OH)2D3 possessed anti-growth effect on human CCA cells with MART-10 much more potent than 1α,25(OH)2D3. The growth inhibition of both drugs were mediated by induction of G0/G1 cell cycle arrest through upregulation of p27 and downregulation of CDK4, CDK6, and cyclin D3. Human neutrophil gelatinase associated lipocalin (NGAL) was found to be involved in 1α,25(OH)2D3 and MART-10 meditated growth inhibition for CCA as knockdown of NGAL decreased Ki-67 expression in SNU308 cells and rendered SNU308 cells less responsive to 1α,25(OH)2D3 and MART-10 treatment. Vitamin D receptor (VDR) knockdown partly abolished MART-10-induced inhibition of NGAL and cell growth in SNU308 cells. The xenograft animal study demonstrated MART-10 could effectively repressed CCA growth in vivo without inducing obvious side effects. The IHC examination of human CCA specimen for VDR revealed that higher VDR expression was linked with better prognosis. Collectively, our results suggest that MART-10 could be a promising regimen for CCA treatment. PMID:28256614

Defective transport of the obesity mutant PC1/3 N222D contributes to loss of function.

PubMed

Prabhu, Yogikala; Blanco, Elias H; Liu, Ming; Peinado, Juan R; Wheeler, Matthew C; Gekakis, Nicholas; Arvan, Peter; Lindberg, Iris

2014-07-01

Mutations in the PCSK1 gene encoding prohormone convertase 1/3 (PC1/3) are strongly associated with obesity in humans. The PC1/3(N222D) mutant mouse thus far represents the only mouse model that mimics the PC1/3 obesity phenotype in humans. The present investigation addresses the cell biology of the N222D mutation. Metabolic labeling experiments reveal a clear defect in the kinetics of insulin biosynthesis in islets from PC1/3(N222D) mutant mice, resulting in an increase in both proinsulin and its processing intermediates, predominantly lacking cleavage at the Arg-Arg site. Although the mutant PC1/3 zymogen is correctly processed to the 87-kDa form, pulse-chase immunoprecipitation experiments, labeling, and immunohistochemical experiments using uncleavable variants all demonstrate that the PC1/3-N222D protein is largely mislocalized compared with similar wild-type (WT) constructs, being predominantly retained in the endoplasmic reticulum. The PC1/3-N222D mutant also undergoes more efficient degradation via the ubiquitin-proteasome system than the WT enzyme. Lastly, the mutant PC1/3-N222D protein coimmunoprecipitates with WT PC1/3 and exerts a modest effect on intracellular retention of the WT enzyme. These profound alterations in the cell biology of PC1/3-N222D are likely to contribute to the defective insulin biosynthetic events observed in the mutant mice and may be relevant to the dramatic contributions of polymorphisms in this gene to human obesity.

Defective Transport of the Obesity Mutant PC1/3 N222D Contributes to Loss of Function

PubMed Central

Prabhu, Yogikala; Blanco, Elias H.; Liu, Ming; Peinado, Juan R.; Wheeler, Matthew C.; Gekakis, Nicholas; Arvan, Peter

2014-01-01

Mutations in the PCSK1 gene encoding prohormone convertase 1/3 (PC1/3) are strongly associated with obesity in humans. The PC1/3N222D mutant mouse thus far represents the only mouse model that mimics the PC1/3 obesity phenotype in humans. The present investigation addresses the cell biology of the N222D mutation. Metabolic labeling experiments reveal a clear defect in the kinetics of insulin biosynthesis in islets from PC1/3N222D mutant mice, resulting in an increase in both proinsulin and its processing intermediates, predominantly lacking cleavage at the Arg-Arg site. Although the mutant PC1/3 zymogen is correctly processed to the 87-kDa form, pulse-chase immunoprecipitation experiments, labeling, and immunohistochemical experiments using uncleavable variants all demonstrate that the PC1/3-N222D protein is largely mislocalized compared with similar wild-type (WT) constructs, being predominantly retained in the endoplasmic reticulum. The PC1/3-N222D mutant also undergoes more efficient degradation via the ubiquitin-proteasome system than the WT enzyme. Lastly, the mutant PC1/3-N222D protein coimmunoprecipitates with WT PC1/3 and exerts a modest effect on intracellular retention of the WT enzyme. These profound alterations in the cell biology of PC1/3-N222D are likely to contribute to the defective insulin biosynthetic events observed in the mutant mice and may be relevant to the dramatic contributions of polymorphisms in this gene to human obesity. PMID:24828610

21 CFR 172.380 - Vitamin D3.

Code of Federal Regulations, 2010 CFR

2010-04-01

... CONSUMPTION (CONTINUED) FOOD ADDITIVES PERMITTED FOR DIRECT ADDITION TO FOOD FOR HUMAN CONSUMPTION Special Dietary and Nutritional Additives § 172.380 Vitamin D3. Vitamin D3 may be used safely in foods as a... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Vitamin D3. 172.380 Section 172.380 Food and Drugs...

Using Computer-Aided Design Software and 3D Printers to Improve Spatial Visualization

ERIC Educational Resources Information Center

Katsio-Loudis, Petros; Jones, Millie

2015-01-01

Many articles have been published on the use of 3D printing technology. From prefabricated homes and outdoor structures to human organs, 3D printing technology has found a niche in many fields, but especially education. With the introduction of AutoCAD technical drawing programs and now 3D printing, learners can use 3D printed models to develop…

3D hybrid electrode structure as implantable interface for a vestibular neural prosthesis in humans.

PubMed

Hoffmann, Klaus-P; Poppendieck, Wigand; Tätzner, Simon; DiGiovanna, Jack; Kos, Maria Izabel; Guinand, Nils; Guyot, Jean-P; Micera, Silvestro

2011-01-01

Implantable interfaces are essential components of vestibular neural prostheses. They interface the biological system with electrical stimulation that is used to restore transfer of vestibular information. Regarding the anatomical situation special 3D structures are required. In this paper, the design and the manufacturing process of a novel 3D hybrid microelectrode structure as interface to the human vestibular system are described. Photolithography techniques, assembling technology and rapid prototyping are used for manufacturing.

Virtual embryology: a 3D library reconstructed from human embryo sections and animation of development process.

PubMed

Komori, M; Miura, T; Shiota, K; Minato, K; Takahashi, T

1995-01-01

The volumetric shape of a human embryo and its development is hard to comprehend as they have been viewed as a 2D schemes in a textbook or microscopic sectional image. In this paper, a CAI and research support system for human embryology using multimedia presentation techniques is described. In this system, 3D data is acquired from a series of sliced specimens. Its 3D structure can be viewed interactively by rotating, extracting, and truncating its whole body or organ. Moreover, the development process of embryos can be animated using a morphing technique applied to the specimen in several stages. The system is intended to be used interactively, like a virtual reality system. Hence, the system is called Virtual Embryology.

Research Summary 3-D Computational Fluid Dynamics (CFD) Model Of The Human Respiratory System

EPA Science Inventory

The U.S. EPA’s Office of Research and Development (ORD) has developed a 3-D computational fluid dynamics (CFD) model of the human respiratory system that allows for the simulation of particulate based contaminant deposition and clearance, while being adaptable for age, ethnicity,...

Synthesis, metabolism, and biological activity of 2-[3-(tetrazolyl)propyl]-1α,25-dihydroxy-19-norvitamin D3.

PubMed

Takano, Masashi; Yasuda, Kaori; Higuchi, Erika; Tohyama, Eri; Takeuchi, Akiko; Sakaki, Toshiyuki; Kittaka, Atsushi

2016-11-01

Recently, we found that 2α-[2-(tetrazol-2-yl)ethyl]-1α,25-dihydroxyvitamin D 3 showed higher osteocalcin promoter transactivation activity in human osteosarcoma (HOS) cells and a greater therapeutic effect in ovariectomized (OVX) rats in vivo than those of active vitamin D 3 , 1α,25(OH) 2 D 3 . We were interested in introducing a heterocyclic ring to the C2 position of the seco-steroidal structure via an alkyl linker, and four novel C2-(3-tetrazolylpropyl) substituted 1α,25-dihydroxy-19-norvitamin D 3 analogs, 2α-[3-(tetrazol-1-yl)propyl]-, 2β-[3-(tetrazol-1-yl)propyl]-, 2α-[3-(tetrazol-2-yl)propyl]-, and 2β-[3-(tetrazol-2-yl)propyl]-19-nor-1α,25(OH) 2 D 3 were synthesized. Among them, 2α-[3-(tetrazol-1-yl)propyl]-19-nor-1α,25(OH) 2 D 3 showed weak binding affinity for human vitamin D receptor (hVDR) (2.6% of 1α,25(OH) 2 D 3 and ca. 15% of 19-nor-1α,25(OH) 2 D 3 ) and weak VDR transactivation activity in HOS cells (EC 50 7.3nM, when 1α,25(OH) 2 D 3 0.23nM). Although the other three compounds could not act as VDR binders by evaluation of the competition assays, 2α-[3-(tetrazol-2-yl)propyl]-19-nor-1α,25(OH) 2 D 3 showed weak transactivation activity (EC 50 12.5nM). Metabolic stability of the 2α-substituted compounds 2α-[3-(tetrazol-1-yl)propyl]- and 2α-[3-(tetrazol-2-yl)propyl]-19-nor-1α,25(OH) 2 D 3 was higher than that of the 2β-substituted counterparts 2β-[3-(tetrazol-1-yl)propyl]- and 2β-[3-(tetrazol-2-yl)propyl]-19-nor-1α,25(OH) 2 D 3 against human CYP24A1. Introduction of a tetrazole ring to the C2-position of the 19-norvitamin D 3 skeleton with the propyl linker led to weak VDR agonistic activity with stability against CYP24A1 metabolism. Copyright © 2015 Elsevier Ltd. All rights reserved.

DHM simulation in virtual environments: a case-study on control room design.

PubMed

Zamberlan, M; Santos, V; Streit, P; Oliveira, J; Cury, R; Negri, T; Pastura, F; Guimarães, C; Cid, G

2012-01-01

This paper will present the workflow developed for the application of serious games in the design of complex cooperative work settings. The project was based on ergonomic studies and development of a control room among participative design process. Our main concerns were the 3D human virtual representation acquired from 3D scanning, human interaction, workspace layout and equipment designed considering ergonomics standards. Using Unity3D platform to design the virtual environment, the virtual human model can be controlled by users on dynamic scenario in order to evaluate the new work settings and simulate work activities. The results obtained showed that this virtual technology can drastically change the design process by improving the level of interaction between final users and, managers and human factors team.

Development of real-time motion capture system for 3D on-line games linked with virtual character

NASA Astrophysics Data System (ADS)

Kim, Jong Hyeong; Ryu, Young Kee; Cho, Hyung Suck

2004-10-01

Motion tracking method is being issued as essential part of the entertainment, medical, sports, education and industry with the development of 3-D virtual reality. Virtual human character in the digital animation and game application has been controlled by interfacing devices; mouse, joysticks, midi-slider, and so on. Those devices could not enable virtual human character to move smoothly and naturally. Furthermore, high-end human motion capture systems in commercial market are expensive and complicated. In this paper, we proposed a practical and fast motion capturing system consisting of optic sensors, and linked the data with 3-D game character with real time. The prototype experiment setup is successfully applied to a boxing game which requires very fast movement of human character.

In vitro pharmacology of aripiprazole, its metabolite and experimental dopamine partial agonists at human dopamine D2 and D3 receptors.

PubMed

Tadori, Yoshihiro; Forbes, Robert A; McQuade, Robert D; Kikuchi, Tetsuro

2011-10-15

Aripiprazole is the first dopamine D(2)/D(3) receptor partial agonist successfully developed and ultimately approved for treatment of a broad spectrum of psychiatric and neurological disorders. Aripiprazole's dopamine D(2) and serotonin 5-HT(1A) receptor partial agonist activities have been postulated to confer clinical efficacy without marked sedation, and a relatively favorable overall side-effect profile. Using aripiprazole's unique profile as a benchmark for new dopamine partial agonist development may facilitate discovery of new antipsychotics. We conducted an in vitro comparative analysis between aripiprazole, and its human metabolite OPC-14857 (7-(4-[4-(2,3-dichlorophenyl)-1-piperazinyl)butoxy)-2(1H)-quinolinone)); RGH-188 (trans-1-[4-[2-[4-(2,3-dichlorophenyl)piperazine-1-yl]ethyl]cyclohexyl]-3,3-dimethylurea), and its metabolite didesmethyl-RGH-188 (DDM-RGH-188); as well as bifeprunox, sarizotan, N-desmethylclozapine (NDMC; clozapine metabolite), and SDZ 208-912 (N-[(8α)-2-chloro-6-methylergolin-8-yl]-2,2-dimethylpropanamide). In vitro pharmacological assessment included inhibition of forskolin-stimulated cAMP accumulation and the reversal of dopamine-induced inhibition in clonal Chinese hamster ovary cell lines expressing D(2S), D(2L), D(3) Ser-9 and D(3) Gly-9 for human dopamine receptors. All test compounds behaved as dopamine D(2)/D(3) receptor partial agonists. Aripiprazole's intrinsic activity at dopamine D(2S) and D(2L) receptors was similar to that of OPC-14857 and RGH-188; lower than that of dopamine and bifeprunox; and higher than that of DDM-RGH-188, SDZ 208-912, sarizotan, and NDMC. Aripiprazole's intrinsic activity at dopamine D(3) Ser-9 and D(3) Gly-9 receptors was similar to that of OPC-14857 and sarizotan; lower than that of dopamine, bifeprunox, RGH-188 and DDM-RGH-188; and higher than that of SDZ 208-912 and NDMC. A consolidated assessment of these findings may help defining the most appropriate magnitude of intrinsic activity at dopamine D(2)/D(3) receptors for clinical efficacy and safety. Copyright © 2011 Elsevier B.V. All rights reserved.

The role of 3-D interactive visualization in blind surveys of H I in galaxies

NASA Astrophysics Data System (ADS)

Punzo, D.; van der Hulst, J. M.; Roerdink, J. B. T. M.; Oosterloo, T. A.; Ramatsoku, M.; Verheijen, M. A. W.

2015-09-01

Upcoming H I surveys will deliver large datasets, and automated processing using the full 3-D information (two positional dimensions and one spectral dimension) to find and characterize H I objects is imperative. In this context, visualization is an essential tool for enabling qualitative and quantitative human control on an automated source finding and analysis pipeline. We discuss how Visual Analytics, the combination of automated data processing and human reasoning, creativity and intuition, supported by interactive visualization, enables flexible and fast interaction with the 3-D data, helping the astronomer to deal with the analysis of complex sources. 3-D visualization, coupled to modeling, provides additional capabilities helping the discovery and analysis of subtle structures in the 3-D domain. The requirements for a fully interactive visualization tool are: coupled 1-D/2-D/3-D visualization, quantitative and comparative capabilities, combined with supervised semi-automated analysis. Moreover, the source code must have the following characteristics for enabling collaborative work: open, modular, well documented, and well maintained. We review four state of-the-art, 3-D visualization packages assessing their capabilities and feasibility for use in the case of 3-D astronomical data.

A High-Calcium and Phosphate Rescue Diet and VDR-Expressing Transgenes Normalize Serum Vitamin D Metabolite Profiles and Renal Cyp27b1 and Cyp24a1 Expression in VDR Null Mice

PubMed Central

Kaufmann, Martin; Lee, Seong Min; Pike, J. Wesley

2015-01-01

Vitamin D receptor (VDR)-mediated 1,25-dihydroxyvitamin D3 (1,25(OH)2D3)-dependent gene expression is compromised in the VDR null mouse. The biological consequences include: hypocalcemia, hypophosphatemia, elevated parathyroid hormone (PTH) and 1,25(OH)2D3, and consequential skeletal abnormalities. CYP24A1 is a cytochrome P450 enzyme that is involved in the side chain oxidation and destruction of both 1,25(OH)2D3 and 25-hydroxyvitamin D3 (25-OH-D3). In the current studies, we used liquid chromatography-tandem mass spectrometry technology to compare the metabolic profiles of VDR null mice fed either a normal or a calcium and phosphate-enriched rescue diet and to assess the consequence of transgenic expression of either mouse or human VDR genes in the same background. Serum 1,25(OH)2D3 levels in VDR null mice on normal chow were highly elevated (>3000 pg/mL) coincident with undetectable levels of catabolites such as 24,25-(OH)2D3 and 25-OH-D3-26,23-lactone normally observed in wild-type mice. The rescue diet corrected serum Ca++, PTH, and 1,25(OH)2D3 values and restored basal expression of Cyp24a1 as evidenced by both renal expression of Cyp24a1 and detection of 24,25-(OH)2D3 and the 25-OH-D3-26,23-lactone. Unexpectedly, this diet also resulted in supranormal levels of 3-epi-24,25-(OH)2D3 and 3-epi-25-OH-D3-26,23-lactone. The reappearance of serum 24,25-(OH)2D3 and renal Cyp24a1 expression after rescue suggests that basal levels of Cyp24a1 may be repressed by high PTH. Introduction of transgenes for either mouse or human VDR also normalized vitamin D metabolism in VDR null mice, whereas this metabolic pattern was unaffected by a transgene encoding a ligand binding-deficient mutant (L233S) human VDR. We conclude that liquid chromatography-tandem mass spectrometry-based metabolic profiling is an ideal analytical method to study mouse models with alterations in calcium/phosphate homeostasis. PMID:26441239

Possible involvement of pregnane X receptor–enhanced CYP24 expression in drug-induced osteomalacia

PubMed Central

Pascussi, Jean Marc; Robert, Agnes; Nguyen, Minh; Walrant-Debray, Odile; Garabedian, Michèle; Martin, Pascal; Pineau, Thierry; Saric, Jean; Navarro, Fréderic; Maurel, Patrick; Vilarem, Marie Josè

2005-01-01

Vitamin D controls calcium homeostasis and the development and maintenance of bones through vitamin D receptor activation. Prolonged therapy with rifampicin or phenobarbital has been shown to cause vitamin D deficiency or osteomalacia, particularly in patients with marginal vitamin D stores. However, the molecular mechanism of this process is unknown. Here we show that these drugs lead to the upregulation of 25-hydroxyvitamin D3-24-hydroxylase (CYP24) gene expression through the activation of the nuclear receptor pregnane X receptor (PXR; NR1I2). CYP24 is a mitochondrial enzyme responsible for inactivating vitamin D metabolites. CYP24 mRNA is upregulated in vivo in mice by pregnenolone 16α-carbonitrile and dexamethasone, 2 murine PXR agonists, and in vitro in human hepatocytes by rifampicin and hyperforin, 2 human PXR agonists. Moreover, rifampicin increased 24-hydroxylase activity in these cells, while, in vivo in mice, pregnenolone 16α-carbonitrile increased the plasma concentration of 24,25-dihydroxyvitamin D3. Transfection of PXR in human embryonic kidney cells resulted in rifampicin-mediated induction of CYP24 mRNA. Analysis of the human CYP24 promoter showed that PXR transactivates the sequence between –326 and –142. We demonstrated that PXR binds to and transactivates the 2 proximal vitamin D–responsive elements of the human CYP24 promoter. These data suggest that xenobiotics and drugs can modulate CYP24 gene expression and alter vitamin D3 hormonal activity and calcium homeostasis through the activation of PXR. PMID:15630458

Metabolism of D-[1-3H]glucose, D-[2-3H]glucose, D-[5-3H]glucose, D-[6-3H]glucose and D-[U-14C]glucose by rat and human erythrocytes incubated in the presence of H2O or D2O.

PubMed

Conget, I; Malaisse, W J

1995-02-01

The present study investigates whether heavy water affects the efficiency of 3HOH production from D-[1-3H]glucose, D-[2-3H]glucose, D-[5-3H]glucose and D-[6-3H]glucose relative to the total generation of tritiated metabolites produced by either rat or human erythrocytes. The relative 3HOH yield was close to 95% with D-[5-3H]glucose, 72% with D-[2-3H]glucose, 22-32% with D-[1-3H]glucose, and only 12% with D-[6-3H]glucose. In the latter case, the comparison of the specific radioactivity of intracellular and extracellular acidic metabolites, expressed relative to that of 14C-labelled metabolites produced from D-[U-14C]glucose, indicated that the generation of 3HOH from D-[6-3H]glucose occurs at distal metabolic steps, such as the partial reversion of the pyruvate kinase reaction or the interconversion of pyruvate and L-alanine in the reaction catalysed by glutamate-pyruvate transaminase. As a rule, the substitution of H2O by D2O only caused minor to negligible changes in the relative 3HOH yield. This implies that the unexpectedly high deuteration of 13C-labelled D-glucose metabolites recently documented in erythrocytes exposed to D2O cannot be attributed to any major interference of heavy water with factors regulating both the deuteration and detritiation efficiency, such as the enzyme-to-enzyme tunnelling of specific glycolytic intermediates.

Protein corona of airborne nanoscale PM2.5 induces aberrant proliferation of human lung fibroblasts based on a 3D organotypic culture.

PubMed

Li, Yan; Wang, Pengcheng; Hu, Chuanlin; Wang, Kun; Chang, Qing; Liu, Lieju; Han, Zhenggang; Shao, Yang; Zhai, Ying; Zuo, Zhengyu; Mak, Michael; Gong, Zhiyong; Wu, Yang

2018-01-31

Exposure to PM2.5 has become one of the most important factors affecting public health in the world. Both clinical and research studies have suggested that PM2.5 inhalation is associated with impaired lung function. In this study, material characterization identified the existence of nanoscale particulate matter (NPM) in airborne PM2.5 samples. When coming into contact with protein-rich fluids, the NPM becomes covered by a protein layer that forms a "protein corona". Based on a 3D organotypic cell culture, the protein corona was shown to mitigate NPM cytotoxicity and further stimulate the proliferation of human lung fibroblasts (HLFs). ROS-activated alpha-smooth muscle actin (α-SMA) is considered to be one of the proliferation pathways. In this research, 3D cell cultures exhibited more tissue-like properties compared with the growth in 2D models. Animal models have been widely used in toxicological research. However, species differences make it impossible to directly translate discoveries from animals to humans. In this research, the 3D HLF model could partly simulate the biological responses of NPM-protein corona-induced aberrant HLF proliferation in the human lung. Our 3D cellular results provide auxiliary support for an animal model in research on PM2.5-induced impaired lung function, particularly in lung fibrosis.

Melanin Transfer in Human 3D Skin Equivalents Generated Exclusively from Induced Pluripotent Stem Cells.

PubMed

Gledhill, Karl; Guo, Zongyou; Umegaki-Arao, Noriko; Higgins, Claire A; Itoh, Munenari; Christiano, Angela M

2015-01-01

The current utility of 3D skin equivalents is limited by the fact that existing models fail to recapitulate the cellular complexity of human skin. They often contain few cell types and no appendages, in part because many cells found in the skin are difficult to isolate from intact tissue and cannot be expanded in culture. Induced pluripotent stem cells (iPSCs) present an avenue by which we can overcome this issue due to their ability to be differentiated into multiple cell types in the body and their unlimited growth potential. We previously reported generation of the first human 3D skin equivalents from iPSC-derived fibroblasts and iPSC-derived keratinocytes, demonstrating that iPSCs can provide a foundation for modeling a complex human organ such as skin. Here, we have increased the complexity of this model by including additional iPSC-derived melanocytes. Epidermal melanocytes, which are largely responsible for skin pigmentation, represent the second most numerous cell type found in normal human epidermis and as such represent a logical next addition. We report efficient melanin production from iPSC-derived melanocytes and transfer within an entirely iPSC-derived epidermal-melanin unit and generation of the first functional human 3D skin equivalents made from iPSC-derived fibroblasts, keratinocytes and melanocytes.

Calcium and 1,25-dihydroxyvitamin D3 modulate genes of immune and inflammatory pathways in the human colon: a human crossover trial123

PubMed Central

Protiva, Petr; Pendyala, Swaroop; Nelson, Celeste; Augenlicht, Leonard H; Lipkin, Martin; Holt, Peter R

2016-01-01

Background: A high dietary calcium intake with adequate vitamin D status has been linked to lower colorectal cancer risk, but the mechanisms of these effects are poorly understood. Objective: The objective of this study was to elucidate the effects of a Western-style diet (WD) and supplemental calcium and/or 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on the colorectal mucosa. Design: We conducted 2 crossover trials to define molecular pathways in the human colorectum altered by 1) a 4-wk WD supplemented with and without 2 g calcium carbonate/d and 2) a 4-wk WD supplemented with 1,25(OH)2D3 (0.5 μg/d) with or without 2 g calcium carbonate/d. The primary study endpoint was genome-wide gene expression in biopsy specimens of the rectosigmoid colonic mucosa. Serum and urinary calcium concentrations were also measured. Results: Changes in urinary calcium accurately reflected calcium consumption. The WD induced modest upregulation of genes involved in inflammatory pathways, including interferon signaling, and calcium supplementation reversed these toward baseline. In contrast, supplementation of the WD with 1,25(OH)2D3 induced striking upregulation of genes involved in inflammation, immune response, extracellular matrix, and cell adhesion. Calcium supplementation largely abrogated these changes. Conclusions: Supplementing 1,25(OH)2D3 to a WD markedly upregulated genes in immune response and inflammation pathways, which were largely reversed by calcium supplementation. This study provides clinical trial evidence of global gene expression changes occurring in the human colorectum in response to calcium and 1,25(OH)2D3 intervention. One action of 1,25(OH)2D3 is to upregulate adaptive immunity. Calcium appears to modulate this effect, pointing to its biological interaction in the mucosa. This trial was registered at clinicaltrials.gov as NCT00298545. Trial protocol is available at http://clinicalstudies.rucares.org (protocol numbers PHO475 and PHO554). PMID:27009752

3D FISH to analyse gene domain-specific chromatin re-modeling in human cancer cell lines.

PubMed

Kocanova, Silvia; Goiffon, Isabelle; Bystricky, Kerstin

2018-06-01

Fluorescence in situ hybridization (FISH) is a common technique used to label DNA and/or RNA for detection of a genomic region of interest. However, the technique can be challenging, in particular when applied to single genes in human cancer cells. Here, we provide a step-by-step protocol for analysis of short (35 kb-300 kb) genomic regions in three dimensions (3D). We discuss the experimental design and provide practical considerations for 3D imaging and data analysis to determine chromatin folding. We demonstrate that 3D FISH using BACs (Bacterial Artificial Chromosomes) or fosmids can provide detailed information of the architecture of gene domains. More specifically, we show that mapping of specific chromatin landscapes informs on changes associated with estrogen stimulated gene activity in human breast cancer cell lines. Copyright © 2018 Elsevier Inc. All rights reserved.

Accuracy of volume measurement using 3D ultrasound and development of CT-3D US image fusion algorithm for prostate cancer radiotherapy.

PubMed

Baek, Jihye; Huh, Jangyoung; Kim, Myungsoo; Hyun An, So; Oh, Yoonjin; Kim, DongYoung; Chung, Kwangzoo; Cho, Sungho; Lee, Rena

2013-02-01

To evaluate the accuracy of measuring volumes using three-dimensional ultrasound (3D US), and to verify the feasibility of the replacement of CT-MR fusion images with CT-3D US in radiotherapy treatment planning. Phantoms, consisting of water, contrast agent, and agarose, were manufactured. The volume was measured using 3D US, CT, and MR devices. A CT-3D US and MR-3D US image fusion software was developed using the Insight Toolkit library in order to acquire three-dimensional fusion images. The quality of the image fusion was evaluated using metric value and fusion images. Volume measurement, using 3D US, shows a 2.8 ± 1.5% error, 4.4 ± 3.0% error for CT, and 3.1 ± 2.0% error for MR. The results imply that volume measurement using the 3D US devices has a similar accuracy level to that of CT and MR. Three-dimensional image fusion of CT-3D US and MR-3D US was successfully performed using phantom images. Moreover, MR-3D US image fusion was performed using human bladder images. 3D US could be used in the volume measurement of human bladders and prostates. CT-3D US image fusion could be used in monitoring the target position in each fraction of external beam radiation therapy. Moreover, the feasibility of replacing the CT-MR image fusion to the CT-3D US in radiotherapy treatment planning was verified.

Activated human primary NK cells efficiently kill colorectal cancer cells in 3D spheroid cultures irrespectively of the level of PD-L1 expression.

PubMed

Lanuza, Pilar M; Vigueras, Alan; Olivan, Sara; Prats, Anne C; Costas, Santiago; Llamazares, Guillermo; Sanchez-Martinez, Diego; Ayuso, José María; Fernandez, Luis; Ochoa, Ignacio; Pardo, Julián

2018-01-01

Haploidentical Natural Killer (NK) cells have been shown as an effective and safe alternative for the treatment of haematological malignancies with poor prognosis for which traditional therapies are ineffective. In contrast to haematological cancer cells, that mainly grow as single suspension cells, solid carcinomas are characterised by a tridimensional (3D) architecture that provide specific surviving advantages and resistance against chemo- and radiotherapy. However, little is known about the impact of 3D growth on solid cancer immunotherapy especially adoptive NK cell transfer. We have recently developed a protocol to activate ex vivo human primary NK cells using B lymphoblastic cell lines, which generates NK cells able to overcome chemoresistance in haematological cancer cells. Here we have analysed the activity of these allogeneic NK cells against colorectal (CRC) human cell lines growing in 3D spheroid culture and correlated with the expression of some of the main ligands regulating NK cell activity. Our results indicate that activated NK cells efficiently kill colorectal tumour cell spheroids in both 2D and 3D cultures. Notably, although 3D CRC cell cultures favoured the expression of the inhibitory immune checkpoint PD-L1, it did not correlate with increased resistance to NK cells. Finally, we have analysed in detail the infiltration of NK cells in 3D spheroids by microscopy and found that at low NK cell density, cell death is not observed although NK cells are able to infiltrate into the spheroid. In contrast, higher densities promote tumoural cell death before infiltration can be detected. These findings show that highly dense activated human primary NK cells efficiently kill colorectal carcinoma cells growing in 3D cultures independently of PD-L1 expression and suggest that the use of allogeneic activated NK cells could be beneficial for the treatment of colorectal carcinoma.

Novel selective human melanocortin-3 receptor ligands: Use of the 4-amino-1,2,4,5-tetrahydro-2-benzazepin-3-one (Aba) scaffold

PubMed Central

Ballet, Steven; Mayorov, Alexander V.; Cai, Minying; Tymecka, Dagmara; Chandler, Kevin B.; Palmer, Erin S.; Van Rompaey, Karolien; Misicka, Aleksandra; Tourwé, Dirk; Hruby, Victor J.

2008-01-01

In search of new selective antagonists and/or agonists for the human melanocortin receptor subtypes hMC1R to hMC5R to elucidate the specific biological roles of each GPCR, we modified the structures of the superagonist MT-II (Ac-Nle-c[Asp-His-D-Phe-Arg-Trp-Lys]-NH2) and the hMC3R/hMC4R antagonist SHU9119 (Ac-Nle-c[Asp-His-D-Nal(2′)-Arg-Trp-Lys]-NH2) by replacing the His-D-Phe and His-D-Nal(2′) fragments in MT-II and SHU9119, respectively, with Aba-Xxx (4-amino-1,2,4,5-tetrahydro-2-benzazepin-3-one-Xxx) dipeptidomimetics (Xxx = D-Phe/pCl-D-Phe/D-Nal(2′)). Employment of the Aba mimetic yielded novel selective high affinity hMC3R and hMC3R/hMC5R antagonists. PMID:17314042

Studies on the analysis of 25-hydroxyvitamin D{sub 3} by isotope-dilution liquid chromatography–tandem mass spectrometry using enzyme-assisted derivatisation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abdel-Khalik, Jonas, E-mail: J.A.F.A.ABDEL-KHALIK.744116@swansea.ac.uk; Crick, Peter J.; Carter, Graham D.

2014-04-11

Highlights: • New method for the analysis of 25-hydroxyvitamin D{sub 3} exploiting Girard P derivatisation. • Method also applicable to vitamin D{sub 3}, 1α,25- and 24,25-dihydroxyvitamin D{sub 3}. • By modification of the method 3-epi-25-hydroxyvitamin D{sub 3} can also be analysed. - Abstract: The total serum concentration of 25-hydroxyvitamins D (25-hydroxyvitamin D{sub 3} and 25-hydroxyvitamin D{sub 2}) is currently used as an indicator of vitamins D status. Vitamins D insufficiency is claimed to be associated with multiple diseases, thus accurate and precise reference methods for the quantification of 25-hydroxyvitamins D are needed. Here we present a novel enzyme-assisted derivatisation methodmore » for the analysis of vitamins D metabolites in adult serum utilising 25-[26,26,26,27,27,27-{sup 2}H{sub 6}]hydroxyvitamin D{sub 3} as the internal standard. Extraction of 25-hydroxyvitamins D from serum is performed with acetonitrile, which is shown to be more efficient than ethanol. Cholesterol oxidase is used to oxidize the 3β-hydroxy group in the vitamins D metabolites followed by derivatisation of the newly formed 3-oxo group with Girard P reagent. 17β-Hydroxysteroid dehydrogenase type 10 is shown to oxidize selectively the 3α-hydroxy group in the 3α-hydroxy epimer of 25-hydroxyvitamin D{sub 3}. Quantification is achieved by isotope-dilution liquid chromatography–tandem mass spectrometry. Recovery experiments for 25-hydroxyvitamin D{sub 3} performed on adult human serum give recovery of 102–106%. Furthermore in addition to 25-hydroxyvitamin D{sub 3}, 24,25-dihydroxyvitamin D{sub 3} and other uncharacterised dihydroxy metabolites, were detected in adult human serum.« less

Vitamin D and gene networks in human osteoblasts

PubMed Central

van de Peppel, Jeroen; van Leeuwen, Johannes P. T. M.

2014-01-01

Bone formation is indirectly influenced by 1,25-dihydroxyvitamin D3 (1,25D3) through the stimulation of calcium uptake in the intestine and re-absorption in the kidneys. Direct effects on osteoblasts and bone formation have also been established. The vitamin D receptor (VDR) is expressed in osteoblasts and 1,25D3 modifies gene expression of various osteoblast differentiation and mineralization-related genes, such as alkaline phosphatase (ALPL), osteocalcin (BGLAP), and osteopontin (SPP1). 1,25D3 is known to stimulate mineralization of human osteoblasts in vitro, and recently it was shown that 1,25D3 induces mineralization via effects in the period preceding mineralization during the pre-mineralization period. For a full understanding of the action of 1,25D3 in osteoblasts it is important to get an integrated network view of the 1,25D3-regulated genes during osteoblast differentiation and mineralization. The current data will be presented and discussed alluding to future studies to fully delineate the 1,25D3 action in osteoblast. Describing and understanding the vitamin D regulatory networks and identifying the dominant players in these networks may help develop novel (personalized) vitamin D-based treatments. The following topics will be discussed in this overview: (1) Bone metabolism and osteoblasts, (2) Vitamin D, bone metabolism and osteoblast function, (3) Vitamin D induced transcriptional networks in the context of osteoblast differentiation and bone formation. PMID:24782782

Genotoxic Effects of Low- and High-LET Radiation on Human Epithelial Cells Grown in 2-D Versus 3-D Culture

NASA Technical Reports Server (NTRS)

Patel, Z. S.; Cucinotta, F. A.; Huff, J. L.

2011-01-01

Risk estimation for radiation-induced cancer relies heavily on human epidemiology data obtained from terrestrial irradiation incidents from sources such as medical and occupational exposures as well as from the atomic bomb survivors. No such data exists for exposures to the types and doses of high-LET radiation that will be encountered during space travel; therefore, risk assessment for space radiation requires the use of data derived from cell culture and animal models. The use of experimental models that most accurately replicate the response of human tissues is critical for precision in risk projections. This work compares the genotoxic effects of radiation on normal human epithelial cells grown in standard 2-D monolayer culture compared to 3-D organotypic co-culture conditions. These 3-D organotypic models mimic the morphological features, differentiation markers, and growth characteristics of fully-differentiated normal human tissue and are reproducible using defined components. Cultures were irradiated with 2 Gy low-LET gamma rays or varying doses of high-LET particle radiation and genotoxic damage was measured using a modified cytokinesis block micronucleus assay. Our results revealed a 2-fold increase in residual damage in 2 Gy gamma irradiated cells grown under organotypic culture conditions compared to monolayer culture. Irradiation with high-LET particle radiation gave similar results, while background levels of damage were comparable under both scenarios. These observations may be related to the phenomenon of "multicellular resistance" where cancer cells grown as 3-D spheroids or in vivo exhibit an increased resistance to killing by chemotherapeutic agents compared to the same cells grown in 2-D culture. A variety of factors are likely involved in mediating this process, including increased cell-cell communication, microenvironment influences, and changes in cell cycle kinetics that may promote survival of damaged cells in 3-D culture that would otherwise die or be rendered reproductively inactive in 2-D culture.

Noninvasive, three-dimensional full-field body sensor for surface deformation monitoring of human body in vivo

NASA Astrophysics Data System (ADS)

Chen, Zhenning; Shao, Xinxing; He, Xiaoyuan; Wu, Jialin; Xu, Xiangyang; Zhang, Jinlin

2017-09-01

Noninvasive, three-dimensional (3-D), full-field surface deformation measurements of the human body are important for biomedical investigations. We proposed a 3-D noninvasive, full-field body sensor based on stereo digital image correlation (stereo-DIC) for surface deformation monitoring of the human body in vivo. First, by applying an improved water-transfer printing (WTP) technique to transfer optimized speckle patterns onto the skin, the body sensor was conveniently and harmlessly fabricated directly onto the human body. Then, stereo-DIC was used to achieve 3-D noncontact and noninvasive surface deformation measurements. The accuracy and efficiency of the proposed body sensor were verified and discussed by considering different complexions. Moreover, the fabrication of speckle patterns on human skin, which has always been considered a challenging problem, was shown to be feasible, effective, and harmless as a result of the improved WTP technique. An application of the proposed stereo-DIC-based body sensor was demonstrated by measuring the pulse wave velocity of human carotid artery.

Effect of 3D-scaffold formation on differentiation and survival in human neural progenitor cells.

PubMed

Ortinau, Stefanie; Schmich, Jürgen; Block, Stephan; Liedmann, Andrea; Jonas, Ludwig; Weiss, Dieter G; Helm, Christiane A; Rolfs, Arndt; Frech, Moritz J

2010-11-11

3D-scaffolds have been shown to direct cell growth and differentiation in many different cell types, with the formation and functionalisation of the 3D-microenviroment being important in determining the fate of the embedded cells. Here we used a hydrogel-based scaffold to investigate the influences of matrix concentration and functionalisation with laminin on the formation of the scaffolds, and the effect of these scaffolds on human neural progenitor cells cultured within them. In this study we used different concentrations of the hydrogel-based matrix PuraMatrix. In some experiments we functionalised the matrix with laminin I. The impact of concentration and treatment with laminin on the formation of the scaffold was examined with atomic force microscopy. Cells from a human fetal neural progenitor cell line were cultured in the different matrices, as well as in a 2D culture system, and were subsequently analysed with antibody stainings against neuronal markers. In parallel, the survival rate of the cells was determined by a live/dead assay. Atomic force microscopy measurements demonstrated that the matrices are formed by networks of isolated PuraMatrix fibres and aggregates of fibres. An increase of the hydrogel concentration led to a decrease in the mesh size of the scaffolds and functionalisation with laminin promoted aggregation of the fibres (bundle formation), which further reduces the density of isolated fibres. We showed that laminin-functionalisation is essential for human neural progenitor cells to build up 3D-growth patterns, and that proliferation of the cells is also affected by the concentration of matrix. In addition we found that 3D-cultures enhanced neuronal differentiation and the survival rate of the cells compared to 2D-cultures. Taken together, we have demonstrated a direct influence of the 3D-scaffold formation on the survival and neuronal differentiation of human neural progenitor cells. These findings emphasize the importance of optimizing 3D-scaffolds protocols prior to in vivo engraftment of stem and progenitor cells in the context of regenerative medicine.

[Comparison of bite marks and teeth features using 2D and 3D methods].

PubMed

Lorkiewicz-Muszyńska, Dorota; Glapiński, Mariusz; Zaba, Czesław; Łabecka, Marzena

2011-01-01

The nature of bite marks is complex. They are found at the scene of crime on different materials and surfaces - not only on human body and corpse, but also on food products and material objects. Human bites on skin are sometimes difficult to interpret and to analyze because of the specific character of skin--elastic and distortable--and because different areas of human body have different surfaces and curvatures. A bite mark left at the scene of crime can be a highly helpful way to lead investigators to criminals. The study was performed to establish: 1) whether bite marks exhibit variations in the accuracy of impressions on different materials, 2) whether it is possible to use the 3D method in the process of identifying an individual based on the comparison of bite marks revealed at the scene, and 3D scans of dental casts, 3) whether application of the 3D method allows for elimination of secondary photographic distortion of bite marks. The authors carried out experiments on simulated cases. Five volunteers bit various materials with different surfaces. Experimental bite marks were collected with emphasis on differentiations of materials. Subsequently, dental impressions were taken from five volunteers in order to prepare five sets of dental casts (the maxilla and mandible. The biting edges of teeth were impressed in wax to create an imprint. The samples of dental casts, corresponding wax bite impressions and bite marks from different materials were scanned with 2D and 3D scanners and photographs were taken. All of these were examined in detail and then compared using different methods (2D and 3D). 1) Bite marks exhibit variations in accuracy of impression on different materials. The most legible reproduction of bite marks was seen on cheese. 2) In comparison of bite marks, the 3D method and 3D scans of dental casts are highly accurate. 3) The 3D method helps to eliminate secondary photographic distortion of bite marks.

Dopamine D3 receptor ligands for drug addiction treatment: update on recent findings.

PubMed

Le Foll, Bernard; Collo, Ginetta; Rabiner, Eugenii A; Boileau, Isabelle; Merlo Pich, Emilio; Sokoloff, Pierre

2014-01-01

The dopamine D3 receptor is located in the limbic area and apparently mediates selective effects on motivation to take drugs and drug-seeking behaviors, so that there has been considerable interest on the possible use of D3 receptor ligands to treat drug addiction. However, only recently selective tools allowing studying this receptor have been developed. This chapter presents an overview of findings that were presented at a symposium on the conference Dopamine 2013 in Sardinia in May 2013. Novel neurobiological findings indicate that drugs of abuse can lead to significant structural plasticity in rodent brain and that this is dependent on the availability of functional dopamine D3 autoreceptor, whose activation increased phosphorylation in the ERK pathway and in the Akt/mTORC1 pathway indicating the parallel engagement of a series of intracellular signaling pathways all involved in cell growth and survival. Preclinical findings using animal models of drug-seeking behaviors confirm that D3 antagonists have a promising profile to treat drug addiction across drugs of abuse type. Imaging the D3 is now feasible in human subjects. Notably, the development of (+)-4-propyl-9-hydroxynaphthoxazine ligand used in positron emission tomography (PET) studies in humans allows to measure D3 and D2 receptors based on the area of the brain under study. This PET ligand has been used to confirm up-regulation of D3 sites in psychostimulant users and to reveal that tobacco smoking produces elevation of dopamine at the level of D3 sites. There are now novel antagonists being developed, but also old drugs such as buspirone, that are available to test the D3 hypothesis in humans. The first results of clinical investigations are now being provided. Overall, those recent findings support further exploration of D3 ligands to treat drug addiction. © 2014 Elsevier B.V. All rights reserved.

3-D Ultrafast Doppler Imaging Applied to the Noninvasive and Quantitative Imaging of Blood Vessels in Vivo

PubMed Central

Provost, J.; Papadacci, C.; Demene, C.; Gennisson, J-L.; Tanter, M.; Pernot, M.

2016-01-01

Ultrafast Doppler Imaging was introduced as a technique to quantify blood flow in an entire 2-D field of view, expanding the field of application of ultrasound imaging to the highly sensitive anatomical and functional mapping of blood vessels. We have recently developed 3-D Ultrafast Ultrasound Imaging, a technique that can produce thousands of ultrasound volumes per second, based on three-dimensional plane and diverging wave emissions, and demonstrated its clinical feasibility in human subjects in vivo. In this study, we show that non-invasive 3-D Ultrafast Power Doppler, Pulsed Doppler, and Color Doppler Imaging can be used to perform quantitative imaging of blood vessels in humans when using coherent compounding of three-dimensional tilted plane waves. A customized, programmable, 1024-channel ultrasound system was designed to perform 3-D Ultrafast Imaging. Using a 32X32, 3-MHz matrix phased array (Vermon, France), volumes were beamformed by coherently compounding successive tilted plane wave emissions. Doppler processing was then applied in a voxel-wise fashion. 3-D Ultrafast Power Doppler Imaging was first validated by imaging Tygon tubes of varying diameter and its in vivo feasibility was demonstrated by imaging small vessels in the human thyroid. Simultaneous 3-D Color and Pulsed Doppler Imaging using compounded emissions were also applied in the carotid artery and the jugular vein in one healthy volunteer. PMID:26276956

Three-dimensional stereotactic atlas of the adult human skull correlated with the brain, cranial nerves, and intracranial vasculature.

PubMed

Nowinski, Wieslaw L; Thaung, Thant Shoon Let; Chua, Beng Choon; Yi, Su Hnin Wut; Ngai, Vincent; Yang, Yili; Chrzan, Robert; Urbanik, Andrzej

2015-05-15

Although the adult human skull is a complex and multifunctional structure, its 3D, complete, realistic, and stereotactic atlas has not yet been created. This work addresses the construction of a 3D interactive atlas of the adult human skull spatially correlated with the brain, cranial nerves, and intracranial vasculature. The process of atlas construction included computed tomography (CT) high-resolution scan acquisition, skull extraction, skull parcellation, 3D disarticulated bone surface modeling, 3D model simplification, brain-skull registration, 3D surface editing, 3D surface naming and color-coding, integration of the CT-derived 3D bony models with the existing brain atlas, and validation. The virtual skull model created is complete with all 29 bones, including the auditory ossicles (being among the smallest bones). It contains all typical bony features and landmarks. The created skull model is superior to the existing skull models in terms of completeness, realism, and integration with the brain along with blood vessels and cranial nerves. This skull atlas is valuable for medical students and residents to easily get familiarized with the skull and surrounding anatomy with a few clicks. The atlas is also useful for educators to prepare teaching materials. It may potentially serve as a reference aid in the reading and operating rooms. Copyright © 2015 Elsevier B.V. All rights reserved.

3-D-Gaze-Based Robotic Grasping Through Mimicking Human Visuomotor Function for People With Motion Impairments.

PubMed

Li, Songpo; Zhang, Xiaoli; Webb, Jeremy D

2017-12-01

The goal of this paper is to achieve a novel 3-D-gaze-based human-robot-interaction modality, with which a user with motion impairment can intuitively express what tasks he/she wants the robot to do by directly looking at the object of interest in the real world. Toward this goal, we investigate 1) the technology to accurately sense where a person is looking in real environments and 2) the method to interpret the human gaze and convert it into an effective interaction modality. Looking at a specific object reflects what a person is thinking related to that object, and the gaze location contains essential information for object manipulation. A novel gaze vector method is developed to accurately estimate the 3-D coordinates of the object being looked at in real environments, and a novel interpretation framework that mimics human visuomotor functions is designed to increase the control capability of gaze in object grasping tasks. High tracking accuracy was achieved using the gaze vector method. Participants successfully controlled a robotic arm for object grasping by directly looking at the target object. Human 3-D gaze can be effectively employed as an intuitive interaction modality for robotic object manipulation. It is the first time that 3-D gaze is utilized in a real environment to command a robot for a practical application. Three-dimensional gaze tracking is promising as an intuitive alternative for human-robot interaction especially for disabled and elderly people who cannot handle the conventional interaction modalities.

Virtual three-dimensional blackboard: three-dimensional finger tracking with a single camera

NASA Astrophysics Data System (ADS)

Wu, Andrew; Hassan-Shafique, Khurram; Shah, Mubarak; da Vitoria Lobo, N.

2004-01-01

We present a method for three-dimensional (3D) tracking of a human finger from a monocular sequence of images. To recover the third dimension from the two-dimensional images, we use the fact that the motion of the human arm is highly constrained owing to the dependencies between elbow and forearm and the physical constraints on joint angles. We use these anthropometric constraints to derive a 3D trajectory of a gesticulating arm. The system is fully automated and does not require human intervention. The system presented can be used as a visualization tool, as a user-input interface, or as part of some gesture-analysis system in which 3D information is important.

What is 3D good for? A review of human performance on stereoscopic 3D displays

NASA Astrophysics Data System (ADS)

McIntire, John P.; Havig, Paul R.; Geiselman, Eric E.

2012-06-01

This work reviews the human factors-related literature on the task performance implications of stereoscopic 3D displays, in order to point out the specific performance benefits (or lack thereof) one might reasonably expect to observe when utilizing these displays. What exactly is 3D good for? Relative to traditional 2D displays, stereoscopic displays have been shown to enhance performance on a variety of depth-related tasks. These tasks include judging absolute and relative distances, finding and identifying objects (by breaking camouflage and eliciting perceptual "pop-out"), performing spatial manipulations of objects (object positioning, orienting, and tracking), and navigating. More cognitively, stereoscopic displays can improve the spatial understanding of 3D scenes or objects, improve memory/recall of scenes or objects, and improve learning of spatial relationships and environments. However, for tasks that are relatively simple, that do not strictly require depth information for good performance, where other strong cues to depth can be utilized, or for depth tasks that lie outside the effective viewing volume of the display, the purported performance benefits of 3D may be small or altogether absent. Stereoscopic 3D displays come with a host of unique human factors problems including the simulator-sickness-type symptoms of eyestrain, headache, fatigue, disorientation, nausea, and malaise, which appear to effect large numbers of viewers (perhaps as many as 25% to 50% of the general population). Thus, 3D technology should be wielded delicately and applied carefully; and perhaps used only as is necessary to ensure good performance.

Three-dimensional ray tracing in spherical and elliptical generalized Luneburg lenses for application in the human eye lens.

PubMed

Gómez-Correa, J E; Coello, V; Garza-Rivera, A; Puente, N P; Chávez-Cerda, S

2016-03-10

Ray tracing in spherical Luneburg lenses has always been represented in 2D. All propagation planes in a 3D spherical Luneburg lens generate the same ray tracing, due to its radial symmetry. A geometry without radial symmetry generates a different ray tracing. For this reason, a new ray tracing method in 3D through spherical and elliptical Luneburg lenses using 2D methods is proposed. The physics of the propagation is shown here, which allows us to make a ray tracing associated with a vortex beam. A 3D ray tracing in a composite modified Luneburg lens that represents the human eye lens is also presented.

Registration Combining Wide and Narrow Baseline Feature Tracking Techniques for Markerless AR Systems.

PubMed

Duan, Liya; Guan, Tao; Yang, Bo

2009-01-01

Augmented reality (AR) is a field of computer research which deals with the combination of real world and computer generated data. Registration is one of the most difficult problems currently limiting the usability of AR systems. In this paper, we propose a novel natural feature tracking based registration method for AR applications. The proposed method has following advantages: (1) it is simple and efficient, as no man-made markers are needed for both indoor and outdoor AR applications; moreover, it can work with arbitrary geometric shapes including planar, near planar and non planar structures which really enhance the usability of AR systems. (2) Thanks to the reduced SIFT based augmented optical flow tracker, the virtual scene can still be augmented on the specified areas even under the circumstances of occlusion and large changes in viewpoint during the entire process. (3) It is easy to use, because the adaptive classification tree based matching strategy can give us fast and accurate initialization, even when the initial camera is different from the reference image to a large degree. Experimental evaluations validate the performance of the proposed method for online pose tracking and augmentation.

A tissue engineered human endometrial stroma that responds to cues for secretory differentiation, decidualization and menstruation

PubMed Central

Schutte, Stacey C.; Taylor, Robert N.

2012-01-01

Objective To show the responsiveness of a tissue engineered human endometrial stroma to combinations of hormones mimicking the secretory and menstrual phases of the cycle. Design In vitro experimental study Setting University uterine biology research laboratory Cells Telomerase immortalized human endometrial stromal cells Interventions The stromal cells were cultured in monolayers (2D) or encapsulated in a collagen I hydrogel (3D) to create a simplified tissue engineered stroma. The cells and tissues were exposed to hormone treatments mimicking early and late secretory phases, decidualization and steroid withdrawal conditions to recapitulate menstruation. Main Outcome Measure(s) Morphological and biochemical markers of decidualization and collagenase activity Result(s) The 3D tissue is capable of manifesting changes in morphology and biochemical markers of decidualization similar to 2D culture and characteristic of endometrial stroma in vivo. Unlike 2D culture, the 3D tissue responded to steroid withdrawal by increased collagenase activity and tissue breakdown. Conclusion(s) 3D tissue engineered endometrial stroma can mimic secretory and menstrual phases of the cycle and may be useful for studying uterine receptivity and menstruation in a physiological endocrine environment. PMID:22306710

High-resolution imaging diagnosis of human fetal membrane by three-dimensional optical coherence tomography

NASA Astrophysics Data System (ADS)

Ren, Hugang; Avila, Cecilia; Kaplan, Cynthia; Pan, Yingtian

2011-11-01

Microscopic chorionic pseudocyst (MCP) arising in the chorion leave of the human fetal membrane (FM) is a clinical precursor for preeclampsia which may progress to fatal medical conditions (e.g., abortion) if left untreated. To examine the utility of three-dimensional (3D) optical coherence tomography (OCT) for noninvasive delineation of the morphology of human fetal membranes and early clinical detection of MCP, 60 human FM specimens were acquired from 10 different subjects undergoing term cesarean delivery for an ex vivo feasibility study. Our results showed that OCT was able to identify the four-layer architectures of human FMs consisting of high-scattering decidua vera (DV, average thickness dDV ~ 92+/-38 μm), low-scattering chorion and trophoblast (CT, dCT ~ 150+/-67 μm), high-scattering subepithelial amnion (A, dA ~ 95+/-36 μm), and low-scattering epithelium (E, dE ~ 29+/-8 μm). Importantly, 3D OCT was able to instantaneously detect MCPs (low scattering due to edema, fluid buildup, vasodilatation) and track (staging) their thicknesses dMCP ranging from 24 to 615 μm. It was also shown that high-frequency ultrasound was able to compliment OCT for detecting more advanced thicker MCPs (e.g., dMCP>615 μm) because of its increased imaging depth.

Highly potent analogues of luteinizing hormone-releasing hormone containing D-phenylalanine nitrogen mustard in position 6.

PubMed Central

Bajusz, S; Janaky, T; Csernus, V J; Bokser, L; Fekete, M; Srkalovic, G; Redding, T W; Schally, A V

1989-01-01

The nitrogen mustard derivatives of 4-phenylbutyric acid and L-phenylalanine, called chlorambucil (Chl) and melphalan (Mel), respectively, have been incorporated into several peptide hormones, including luteinizing hormone-releasing hormone (LH-RH). The alkylating analogues of LH-RH were prepared by linking Chl, as an N-acyl moiety, to the complete amino acid sequence of agonistic and antagonistic analogues. These compounds, in particular the antagonistic analogues, showed much lower potency than their congeners carrying other acyl groups. To obtain highly potent alkylating analogues of LH-RH, the D enantiomer of Mel was incorporated into position 6 of the native hormone and some of its antagonistic analogues. Of the peptides prepared, [D-Mel6]LH-RH (SB-05) and [Ac-D-Nal(2)1,D-Phe(pCl)2,D-Pal(3)3,Arg5,D-Mel6,D-Ala10++ +]LH-RH [SB-86, where Nal(2) is 3-(2-naphthyl)alanine and Pal(3) is 3-(3-pyridyl)alanine] possessed the expected high agonistic and antagonistic activities, respectively, and also showed high affinities for the membrane receptors of rat pituitary cells, human breast cancer cells, human prostate cancer cells, and rat Dunning R-3327 prostate tumor cells. These two analogues exerted cytotoxic effects on human and rat mammary cancer cells in vitro. Thus these two D-Mel6 analogues seem to be particularly suitable for the study of how alkylating analogues of LH-RH could interfere with intracellular events in certain cancer cells. PMID:2548207

Highly potent analogues of luteinizing hormone-releasing hormone containing D-phenylalanine nitrogen mustard in position 6.

PubMed

Bajusz, S; Janaky, T; Csernus, V J; Bokser, L; Fekete, M; Srkalovic, G; Redding, T W; Schally, A V

1989-08-01

The nitrogen mustard derivatives of 4-phenylbutyric acid and L-phenylalanine, called chlorambucil (Chl) and melphalan (Mel), respectively, have been incorporated into several peptide hormones, including luteinizing hormone-releasing hormone (LH-RH). The alkylating analogues of LH-RH were prepared by linking Chl, as an N-acyl moiety, to the complete amino acid sequence of agonistic and antagonistic analogues. These compounds, in particular the antagonistic analogues, showed much lower potency than their congeners carrying other acyl groups. To obtain highly potent alkylating analogues of LH-RH, the D enantiomer of Mel was incorporated into position 6 of the native hormone and some of its antagonistic analogues. Of the peptides prepared, [D-Mel6]LH-RH (SB-05) and [Ac-D-Nal(2)1,D-Phe(pCl)2,D-Pal(3)3,Arg5,D-Mel6,D-Ala10++ +]LH-RH [SB-86, where Nal(2) is 3-(2-naphthyl)alanine and Pal(3) is 3-(3-pyridyl)alanine] possessed the expected high agonistic and antagonistic activities, respectively, and also showed high affinities for the membrane receptors of rat pituitary cells, human breast cancer cells, human prostate cancer cells, and rat Dunning R-3327 prostate tumor cells. These two analogues exerted cytotoxic effects on human and rat mammary cancer cells in vitro. Thus these two D-Mel6 analogues seem to be particularly suitable for the study of how alkylating analogues of LH-RH could interfere with intracellular events in certain cancer cells.

Hepatic differentiation of human iPSCs in different 3D models: A comparative study.

PubMed

Meier, Florian; Freyer, Nora; Brzeszczynska, Joanna; Knöspel, Fanny; Armstrong, Lyle; Lako, Majlinda; Greuel, Selina; Damm, Georg; Ludwig-Schwellinger, Eva; Deschl, Ulrich; Ross, James A; Beilmann, Mario; Zeilinger, Katrin

2017-12-01

Human induced pluripotent stem cells (hiPSCs) are a promising source from which to derive distinct somatic cell types for in vitro or clinical use. Existent protocols for hepatic differentiation of hiPSCs are primarily based on 2D cultivation of the cells. In the present study, the authors investigated the generation of hiPSC-derived hepatocyte-like cells using two different 3D culture systems: A 3D scaffold-free microspheroid culture system and a 3D hollow-fiber perfusion bioreactor. The differentiation outcome in these 3D systems was compared with that in conventional 2D cultures, using primary human hepatocytes as a control. The evaluation was made based on specific mRNA expression, protein secretion, antigen expression and metabolic activity. The expression of α-fetoprotein was lower, while cytochrome P450 1A2 or 3A4 activities were higher in the 3D culture systems as compared with the 2D differentiation system. Cells differentiated in the 3D bioreactor showed an increased expression of albumin and hepatocyte nuclear factor 4α, as well as secretion of α-1-antitrypsin as compared with the 2D differentiation system, suggesting a higher degree of maturation. In contrast, the 3D scaffold-free microspheroid culture provides an easy and robust method to generate spheroids of a defined size for screening applications, while the bioreactor culture model provides an instrument for complex investigations under physiological-like conditions. In conclusion, the present study introduces two 3D culture systems for stem cell derived hepatic differentiation each demonstrating advantages for individual applications as well as benefits in comparison with 2D cultures.

A novel organotypic 3D sweat gland model with physiological functionality

PubMed Central

Grüdl, Sabine; Banowski, Bernhard; Giesen, Melanie; Sättler, Andrea; Proksch, Peter; Welss, Thomas; Förster, Thomas

2017-01-01

Dysregulated human eccrine sweat glands can negatively impact the quality-of-life of people suffering from disorders like hyperhidrosis. Inability of sweating can even result in serious health effects in humans affected by anhidrosis. The underlying mechanisms must be elucidated and a reliable in vitro test system for drug screening must be developed. Here we describe a novel organotypic three-dimensional (3D) sweat gland model made of primary human eccrine sweat gland cells. Initial experiments revealed that eccrine sweat gland cells in a two-dimensional (2D) culture lose typical physiological markers. To resemble the in vivo situation as close as possible, we applied the hanging drop cultivation technology regaining most of the markers when cultured in its natural spherical environment. To compare the organotypic 3D sweat gland model versus human sweat glands in vivo, we compared markers relevant for the eccrine sweat gland using transcriptomic and proteomic analysis. Comparing the marker profile, a high in vitro-in vivo correlation was shown. Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), muscarinic acetylcholine receptor M3 (CHRM3), Na+-K+-Cl- cotransporter 1 (NKCC1), calcium-activated chloride channel anoctamin-1 (ANO1/TMEM16A), and aquaporin-5 (AQP5) are found at significant expression levels in the 3D model. Moreover, cholinergic stimulation with acetylcholine or pilocarpine leads to calcium influx monitored in a calcium flux assay. Cholinergic stimulation cannot be achieved with the sweat gland cell line NCL-SG3 used as a sweat gland model system. Our results show clear benefits of the organotypic 3D sweat gland model versus 2D cultures in terms of the expression of essential eccrine sweat gland key regulators and in the physiological response to stimulation. Taken together, this novel organotypic 3D sweat gland model shows a good in vitro-in vivo correlation and is an appropriate alternative for screening of potential bioactives regulating the sweat mechanism. PMID:28796813

Structured-light 3D scanner in use to assess the human body posture in physical therapy - a pilot study.

PubMed

Kurzydło, Wojciech; Stach, Beata; Bober, Aleksandra; Wodzińska, Mariola; Długosz, Mirosława M

2014-01-01

The main goal of this study was to asses the possibility of using mass production structured-light 3d scanner to asses human body posture. The study was conducted on a healthy 23 year old volunteer and a lay-figure. The experiment consisted of 28 3D scans, divided into three separate tests. The largest deviation observed in the first two trials was 24.42 mm. While the largest deviation observed in the third trial was 49.91 mm. Data obtained with the mass production structured-light 3d scanner may have comparable or better performance than commercially available systems for the assessment of BP.

Physical security and cyber security issues and human error prevention for 3D printed objects: detecting the use of an incorrect printing material

NASA Astrophysics Data System (ADS)

Straub, Jeremy

2017-06-01

A wide variety of characteristics of 3D printed objects have been linked to impaired structural integrity and use-efficacy. The printing material can also have a significant impact on the quality, utility and safety characteristics of a 3D printed object. Material issues can be created by vendor issues, physical security issues and human error. This paper presents and evaluates a system that can be used to detect incorrect material use in a 3D printer, using visible light imaging. Specifically, it assesses the ability to ascertain the difference between materials of different color and different types of material with similar coloration.

A pilot study of the photoprotective effect of almond phytochemicals in a 3D human skin equivalent

USDA-ARS?s Scientific Manuscript database

UV exposure causes oxidative stress, inflammation, erythema, and skin cancer. Alpha-Tocopherol (AT) and polyphenols (AP) present in almonds may serve as photoprotectants. Our objectives were to assess the feasibility of using a 3D human skin equivalent (HSE) in photoprotectant research and to deter...

Reduced bone resorption by intake of dietary vitamin D and K from tailor-made Atlantic salmon: A randomized intervention trial.

PubMed

Graff, Ingvild Eide; Øyen, Jannike; Kjellevold, Marian; Frøyland, Livar; Gjesdal, Clara Gram; Almås, Bjørg; Rosenlund, Grethe; Lie, Øyvind

2016-10-25

Suboptimal vitamin D status is common among humans, and might increase bone resorption with subsequent negative effects on bone health. Fatty fish, including Atlantic salmon, is an important dietary vitamin D source. However, due to a considerable change in fish feed composition, the contribution of vitamin D from salmon fillet has been reduced. The main objective was to investigate if intake of vitamin D3 enriched salmon or vitamin D3 tablets decreased bone biomarkers (urinary N-telopeptides, deoxypyridinoline, serum bone-specific alkaline phosphatase, and osteocalcin) compared to a low vitamin D3 intake. The 122 healthy postmenopausal women included in this 12 weeks intervention trial were randomized into four groups: three salmon groups (150 grams/two times/week) and one tablet group (800 IU vitamin D and 1000 mg calcium/day). The salmon groups also received calcium supplements. The salmon had three different vitamin D3/vitamin K1 combinations: high D3+high K1, low D3+high K1, or high D3+low K1. Increased intake of salmon containing high levels of vitamin D3 (0.35-0.38 mg/kg/fillet) and supplements with the same weekly contribution had a positive influence on bone health as measured by bone biomarkers in postmenopausal women. Consequently, an increased level of vitamin D3 at least to original level in feed for salmonids will contribute to an improved vitamin D3 status and may improve human bone health.

Human neuron-astrocyte 3D co-culture-based assay for evaluation of neuroprotective compounds.

PubMed

Terrasso, Ana Paula; Silva, Ana Carina; Filipe, Augusto; Pedroso, Pedro; Ferreira, Ana Lúcia; Alves, Paula Marques; Brito, Catarina

Central nervous system drug development has registered high attrition rates, mainly due to the lack of efficacy of drug candidates, highlighting the low reliability of the models used in early-stage drug development and the need for new in vitro human cell-based models and assays to accurately identify and validate drug candidates. 3D human cell models can include different tissue cell types and represent the spatiotemporal context of the original tissue (co-cultures), allowing the establishment of biologically-relevant cell-cell and cell-extracellular matrix interactions. Nevertheless, exploitation of these 3D models for neuroprotection assessment has been limited due to the lack of data to validate such 3D co-culture approaches. In this work we combined a 3D human neuron-astrocyte co-culture with a cell viability endpoint for the implementation of a novel in vitro neuroprotection assay, over an oxidative insult. Neuroprotection assay robustness and specificity, and the applicability of Presto Blue, MTT and CytoTox-Glo viability assays to the 3D co-culture were evaluated. Presto Blue was the adequate endpoint as it is non-destructive and is a simpler and reliable assay. Semi-automation of the cell viability endpoint was performed, indicating that the assay setup is amenable to be transferred to automated screening platforms. Finally, the neuroprotection assay setup was applied to a series of 36 test compounds and several candidates with higher neuroprotective effect than the positive control, Idebenone, were identified. The robustness and simplicity of the implemented neuroprotection assay with the cell viability endpoint enables the use of more complex and reliable 3D in vitro cell models to identify and validate drug candidates. Copyright © 2016 Elsevier Inc. All rights reserved.

Human microbiome visualization using 3D technology.

PubMed

Moore, Jason H; Lari, Richard Cowper Sal; Hill, Douglas; Hibberd, Patricia L; Madan, Juliette C

2011-01-01

High-throughput sequencing technology has opened the door to the study of the human microbiome and its relationship with health and disease. This is both an opportunity and a significant biocomputing challenge. We present here a 3D visualization methodology and freely-available software package for facilitating the exploration and analysis of high-dimensional human microbiome data. Our visualization approach harnesses the power of commercial video game development engines to provide an interactive medium in the form of a 3D heat map for exploration of microbial species and their relative abundance in different patients. The advantage of this approach is that the third dimension provides additional layers of information that cannot be visualized using a traditional 2D heat map. We demonstrate the usefulness of this visualization approach using microbiome data collected from a sample of premature babies with and without sepsis.

Symbolic modeling of human anatomy for visualization and simulation

NASA Astrophysics Data System (ADS)

Pommert, Andreas; Schubert, Rainer; Riemer, Martin; Schiemann, Thomas; Tiede, Ulf; Hoehne, Karl H.

1994-09-01

Visualization of human anatomy in a 3D atlas requires both spatial and more abstract symbolic knowledge. Within our 'intelligent volume' model which integrates these two levels, we developed and implemented a semantic network model for describing human anatomy. Concepts for structuring (abstraction levels, domains, views, generic and case-specific modeling, inheritance) are introduced. Model, tools for generation and exploration and applications in our 3D anatomical atlas are presented and discussed.

Adaptive increase in D3 dopamine receptors in the brain reward circuits of human cocaine fatalities.

PubMed

Staley, J K; Mash, D C

1996-10-01

The mesolimbic dopaminergic system plays a primary role in mediating the euphoric and rewarding effects of most abused drugs. Chronic cocaine use is associated with an increase in dopamine neurotransmission resulting from the blockade of dopamine uptake and is mediated by the activation of dopamine receptors. Recent studies have suggested that the D3 receptor subtype plays a pivotal role in the reinforcing effects of cocaine. The D3 receptor-preferring agonist 7-hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) is a reinforcer in rhesus monkeys trained to self-administer cocaine, but not in cocainenaive monkeys. In vitro autoradiographic localization of [3H]-(+)-7-OH-DPAT binding in the human brain demonstrated that D3 receptors were prevalent and highly localized over the ventromedial sectors of the striatum. Pharmacological characterization of [3H]-(+)-7-OH-DPAT binding to the human nucleus accumbens demonstrated a rank order of potency similar to that observed for binding to the cloned D3 receptor expressed in transfected cell lines. Region-of-interest analysis of [3H]-(+)-7-OH-DPAT binding to the D3 receptor demonstrated a one- to threefold elevation in the number of binding sites over particular sectors of the striatum and substantia nigra in cocaine overdose victims as compared with age-matched and drug-free control subjects. The elevated number of [3H]-(+)-7-OH-DPAT binding sites demonstrates that adaptive changes in the D3 receptor in the reward circuitry of the brain are associated with chronic cocaine abuse. These results suggest that the D3 receptor may be a useful target for drug development of anticocaine medications.

1α,25-dihydroxyvitamin D3 acts via transforming growth factor-β to up-regulate expression of immunosuppressive CD73 on human CD4+ Foxp3- T cells.

PubMed

Mann, Elizabeth H; Chambers, Emma S; Chen, Yin-Huai; Richards, David F; Hawrylowicz, Catherine M

2015-11-01

Vitamin D deficiency is associated with increased incidence and severity of various immune-mediated diseases. Active vitamin D (1α,25-dihydroxyvitamin D3; 1,25(OH)2 D3) up-regulates CD4(+) T-cell expression of the purine ectonucleotidase CD39, a molecule that is associated with the generation of anti-inflammatory adenosine. Here we aimed to investigate the direct impact of 1,25(OH)2 D3 on expression of the downstream ecto-5'-nucleotidase CD73 by human CD4 T cells, and components of the transforming growth factor-β (TGF-β) pathway, which have been implicated in the modulation of CD73 by murine T cells. At 10(-8) to 10(-7) m, 1,25(OH)2 D3 significantly increased expression of CD73 on peripheral human CD4(+) T cells. Although 1,25(OH)2 D3 did not affect the mRNA expression of latent TGF-β1 , 1,25(OH)2 D3 did up-regulate expression of TGF-β-associated molecules [latency-associated peptide (LAP), glycophorin A repetitions predominant (GARP), GP96, neuropilin-1, thrombospondin-1 and αv integrin] which is likely to have contributed to the observed enhancement in TGF-β bioactivity. CD73 was highly co-expressed with LAP and GARP following 1,25(OH)2 D3 treatment, but unexpectedly, each of these cell surface molecules was expressed primarily on CD4(+) Foxp3(-) T cells, rather than CD4(+) Foxp3(+) T cells. Notably, neutralization of TGF-β significantly impaired 1,25(OH)2 D3-mediated induction of CD73. Collectively, we show that 1,25(OH)2 D3 enhances expression of CD73 on CD4(+) Foxp3(-) T cells in a process that is at least partially TGF-β-dependent. These data reveal an additional contributing mechanism by which vitamin D may be protective in immune-mediated disease. © 2015 John Wiley & Sons Ltd.

Comfort and pressure distribution in a human contour shaped aircraft seat (developed with 3D scans of the human body).

PubMed

Smulders, M; Berghman, K; Koenraads, M; Kane, J A; Krishna, K; Carter, T K; Schultheis, U

2016-08-12

The concept of comfort is one way for the growing airline market to differentiate and build customer loyalty. This work follows the idea that increasing the contact area between human and seat can have a positive effect on comfort [5, 6, 7]. To improve comfort, reduce weight and optimise space used, a human contour shaped seat shell and cushioning was developed. First the most common activities, the corresponding postures and seat inclination angles were defined. The imprints of these postures on a rescue mat were 3D scanned and an average human contour curve was defined. The outcome was transferred to a prototype seat that was used to test the effect on perceived comfort/discomfort and pressure distribution. The resulting human contour based prototype seat has comfort and discomfort scores comparable to a traditional seat. The prototype seat had a significantly lower average pressure between subjects' buttocks and the seat pan over a traditional seat. This study shows that it is possible to design a seat pan and backrest based on the different contours of study subjects using 3D scan technology. However, translating the 3D scans into a prototype seat also showed that this can only be seen as a first step; additionally biomechanical information and calculations are needed to create ergonomic seats. Furthermore, it is not possible to capture all different human shapes and postures and translate these into one human contour shape that fits all activities and all human sizes.

The involvement of cyclin D1 degradation through GSK3β-mediated threonine-286 phosphorylation-dependent nuclear export in anti-cancer activity of mulberry root bark extracts.

PubMed

Eo, Hyun Ji; Park, Gwang Hun; Jeong, Jin Boo

2016-02-15

Mulberry root bark was shown to induce cyclin D1 proteasomal degradation in the human colorectal cancer cells. Still, the molecular mechanisms whereby mulberry root bark induces cyclin D1 proteasomal degradation remain largely unknown. In this study, the inhibitory effect of mulberry root bark (MRB) on the proliferation of human colorectal cancer cells and the mechanism of action were examined to evaluate its anti-cancer activity. Anti-proliferative effect was determined by MTT assay. RT-PCR and Western blotting were used to assess the mRNA and protein expression of related proteins. MRB inhibited markedly the proliferation of human colorectal cancer cells (HCT116, SW480 and LoVo). In addition, the proliferation of human breast cancer cells (MDA-MB-231 and MCF-7) was suppressed by MRB treatment. However, MRB did not affect the growth of HepG-2 cells as a human hepatocellular carcinoma cell line. MRB effectively decreased cyclin D1 protein level in human colorectal cancer cells and breast cancer cells, but not in hepatocellular carcinoma cells. Contrast to protein levels, cyclin D1 mRNA level did not be changed by MRB treatment. Inhibition of proteasomal degradation by MG132 attenuated MRB-mediated cyclin D1 downregulation and the half-life of cyclin D1 was decreased in the cells treated with MRB. In addition, MRB increased phosphorylation of cyclin D1 at threonine-286 and a point mutation of threonine-286 to alanine attenuated MRB-mediated cyclin D1 degradation. Inhibition of GSK3β by LiCl suppressed cyclin D1 phosphorylation and downregulation by MRB. MRB decreased the nuclear level of cyclin D1 and the inhibition of nuclear export by LMB attenuated MRB-mediated cyclin D1 degradation. MRB has anti-cancer activity by inducing cyclin D1 proteasomal degradation through cyclin D1 nuclear export via GSK3β-dependent threonine-286 phosphorylation. These findings suggest that possibly its extract could be used for treating colorectal cancer. Copyright © 2015 Elsevier GmbH. All rights reserved.

A framework for geometry acquisition, 3-D printing, simulation, and measurement of head-related transfer functions with a focus on hearing-assistive devices

PubMed Central

Harder, Stine; Paulsen, Rasmus R.; Larsen, Martin; Laugesen, Søren; Mihocic, Michael; Majdak, Piotr

2017-01-01

Individual head-related transfer functions (HRTFs) are essential in applications like fitting hearing-assistive devices (HADs) for providing accurate sound localization performance. Individual HRTFs are usually obtained through intricate acoustic measurements. This paper investigates the use of a three-dimensional (3D) head model for acquisition of individual HRTFs. Two aspects were investigated; whether a 3D-printed model can replace measurements on a human listener and whether numerical simulations can replace acoustic measurements. For this purpose, HRTFs were acoustically measured for four human listeners and for a 3D printed head model of one of these listeners. Further, HRTFs were simulated by applying the finite element method to the 3D head model. The monaural spectral features and spectral distortions were very similar between re-measurements and between human and printed measurements, however larger deviations were observed between measurement and simulation. The binaural cues were in agreement among all HRTFs of the same listener, indicating that the 3D model is able to provide localization cues potentially accessible to HAD users. Hence, the pipeline of geometry acquisition, printing, and acoustic measurements or simulations, seems to be a promising step forward towards in-silico design of HADs. PMID:28239188

Translesion synthesis DNA polymerases promote error-free replication through the minor-groove DNA adduct 3-deaza-3-methyladenine.

PubMed

Yoon, Jung-Hoon; Roy Choudhury, Jayati; Park, Jeseong; Prakash, Satya; Prakash, Louise

2017-11-10

N3-Methyladenine (3-MeA) is formed in DNA by reaction with S -adenosylmethionine, the reactive methyl donor, and by reaction with alkylating agents. 3-MeA protrudes into the DNA minor groove and strongly blocks synthesis by replicative DNA polymerases (Pols). However, the mechanisms for replicating through this lesion in human cells remain unidentified. Here we analyzed the roles of translesion synthesis (TLS) Pols in the replication of 3-MeA-damaged DNA in human cells. Because 3-MeA has a short half-life in vitro , we used the stable 3-deaza analog, 3-deaza-3-methyladenine (3-dMeA), which blocks the DNA minor groove similarly to 3-MeA. We found that replication through the 3-dMeA adduct is mediated via three different pathways, dependent upon Polι/Polκ, Polθ, and Polζ. As inferred from biochemical studies, in the Polι/Polκ pathway, Polι inserts a nucleotide (nt) opposite 3-dMeA and Polκ extends synthesis from the inserted nt. In the Polθ pathway, Polθ carries out both the insertion and extension steps of TLS opposite 3-dMeA, and in the Polζ pathway, Polζ extends synthesis following nt insertion by an as yet unidentified Pol. Steady-state kinetic analyses indicated that Polι and Polθ insert the correct nt T opposite 3-dMeA with a much reduced catalytic efficiency and that both Pols exhibit a high propensity for inserting a wrong nt opposite this adduct. However, despite their low fidelity of synthesis opposite 3-dMeA, TLS opposite this lesion replicates DNA in a highly error-free manner in human cells. We discuss the implications of these observations for TLS mechanisms in human cells. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Single oral dose pharmacokinetics of decursin and decursinol angelate in healthy adult men and women.

PubMed

Zhang, Jinhui; Li, Li; Hale, Thomas W; Chee, Wayne; Xing, Chengguo; Jiang, Cheng; Lü, Junxuan

2015-01-01

The ethanol extract of Angelica gigas Nakai (AGN) root has promising anti-cancer and other bioactivities in rodent models. It is currently believed that the pyranocoumarin isomers decursin (D) and decursinol angelate (DA) contribute to these activities. We and others have documented that D and DA were rapidly converted to decursinol (DOH) in rodents. However, our in vitro metabolism studies suggested that D and DA might be metabolized differently in humans. To test this hypothesis and address a key question for human translatability of animal model studies of D and DA or AGN extract, we conducted a single oral dose human pharmacokinetic study of D and DA delivered through an AGN-based dietary supplement Cogni.Q (purchased from Quality of Life Labs, Purchase, NY) in twenty healthy subjects, i.e., 10 men and 10 women, each consuming 119 mg D and 77 mg DA from 4 vegicaps. Analyses of plasma samples using UHPLC-MS/MS showed mean time to peak concentration (Tmax) of 2.1, 2.4 and 3.3 h and mean peak concentration (Cmax) of 5.3, 48.1 and 2,480 nmol/L for D, DA and DOH, respectively. The terminal elimination half-life (t1/2) for D and DA was similar (17.4 and 19.3 h) and each was much longer than that of DOH (7.4 h). The mean area under the curve (AUC0-48h) for D, DA and DOH was estimated as 37, 335 and 27,579 h∙nmol/L, respectively. Gender-wise, men absorbed the parent compounds faster and took shorter time to reach DOH peak concentration. The human data supported an extensive conversion of D and DA to DOH, even though they metabolized DA slightly slower than rodents. Therefore, the data generated in rodent models concerning anti-cancer efficacy, safety, tissue distribution and pharmacodynamic biomarkers will likely be relevant for human translation. ClinicalTrials.gov NCT02114957.

Single Oral Dose Pharmacokinetics of Decursin and Decursinol Angelate in Healthy Adult Men and Women

PubMed Central

Zhang, Jinhui; Li, Li; Hale, Thomas W.; Chee, Wayne; Xing, Chengguo; Jiang, Cheng; Lü, Junxuan

2015-01-01

The ethanol extract of Angelica gigas Nakai (AGN) root has promising anti-cancer and other bioactivities in rodent models. It is currently believed that the pyranocoumarin isomers decursin (D) and decursinol angelate (DA) contribute to these activities. We and others have documented that D and DA were rapidly converted to decursinol (DOH) in rodents. However, our in vitro metabolism studies suggested that D and DA might be metabolized differently in humans. To test this hypothesis and address a key question for human translatability of animal model studies of D and DA or AGN extract, we conducted a single oral dose human pharmacokinetic study of D and DA delivered through an AGN-based dietary supplement Cogni.Q (purchased from Quality of Life Labs, Purchase, NY) in twenty healthy subjects, i.e., 10 men and 10 women, each consuming 119 mg D and 77 mg DA from 4 vegicaps. Analyses of plasma samples using UHPLC-MS/MS showed mean time to peak concentration (Tmax) of 2.1, 2.4 and 3.3 h and mean peak concentration (Cmax) of 5.3, 48.1 and 2,480 nmol/L for D, DA and DOH, respectively. The terminal elimination half-life (t1/2) for D and DA was similar (17.4 and 19.3 h) and each was much longer than that of DOH (7.4 h). The mean area under the curve (AUC0-48h) for D, DA and DOH was estimated as 37, 335 and 27,579 h∙nmol/L, respectively. Gender-wise, men absorbed the parent compounds faster and took shorter time to reach DOH peak concentration. The human data supported an extensive conversion of D and DA to DOH, even though they metabolized DA slightly slower than rodents. Therefore, the data generated in rodent models concerning anti-cancer efficacy, safety, tissue distribution and pharmacodynamic biomarkers will likely be relevant for human translation. Trial Registration ClinicalTrials.gov NCT02114957 PMID:25695490

Host-pathogen-interaction reconstituted in 3-dimensional cocultures of mucosa and C. albicans.

PubMed

Buchs, Romina; Lehner, Bruno; Meuwly, Phillippe; Schnyder, Bruno

2018-06-14

C. albicans frequently causes recurrent intimal infectious disease (ID). This demands the treatment of multiple phases of the infection. The objective of this study was to uncover the host-pathogen-interaction using 2D epithelium cell-barrier and 3D subepithelium tissue cells of human mucosa. The 2D cell cultures assessed C. albicans adhesion. Addition of the anti-fungal drug Fluconazol did not inhibit the adhesion, despite its pathogen growth inhibition (MIC value 0.08μg/mL). A 3D tissue was engineered in multi-transwells by placing human fibroblast cultures on a thick porous scaffold. This contained the yeast placed in the top compartment and prevented passive penetration. After 28h the pathogen transmigrated the barrier and was collected in the bottom compartment. A change in pathogen morphology was observed where hypha formed and grew to be 231μm long after 28h. The hypha was thus long enough to cross the 200μm thick 3D tissue. The 3D infection was inhibited by addition of Fluconazol (0.08μg/mL), confirming that penetration is dependent on pathogen growth. In conclusion, ID was reconstituted step-by-step on 2D epithelium surface and in 3D connective tissue of human mucosa. Fluconazol growth-inhibition of the pathogen C. albicans was confirmed in the 3D tissue. We thus propose that this ID in vitro test is suitable for the identification and characterization of new treatments against C. albicans..

Ultraviolet- and infrared-induced 11 beta-hydroxysteroid dehydrogenase type 1 activating skin photoaging is inhibited by red ginseng extract containing high concentration of ginsenoside Rg3(S).

PubMed

Nam, Jin-Ju; Min, Ji-Eun; Son, Min-Ho; Oh, Jin-Hwan; Kang, Seunghyun

2017-11-01

Sun irradiation is one of major extrinsic stressors responsible for premature skin aging through activation and expression of 11 beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which converts inactive cortisone to active cortisol. The aim of this study was to evaluate the inhibitory effects of red ginseng extract containing high concentrations of ginsenoside Rg3 (S) (GERg3) on 11β-HSD1-induced skin photoaging. To evaluate the inhibitory effects of GERg3 on ultraviolet- (UV) or infrared (IR)-induced skin photoaging, human dermal fibroblasts or a normal human 3D skin model was exposed to UV or an IR. RT-PCR, ELISA, Western blot, and H&E staining were used for evaluations. GERg3 was isolated from crude red ginseng. GERg3 inhibited the increased expressions of 11β-HSD1, interleukin (IL)-6, and matrix metalloproteinase-1 (MMP-1) in UVB- or IR-exposed Hs68 cells. Additionally, the increased cortisol, IL-6, and MMP-1 expressions were effectively reduced by GERg3 in UVA-exposed 3D skin models. The photoinduced decrease in type 1 procollagen also recovered as a result of GERg3 treatment in Hs68 cells and the 3D skin model. In addition, the UVA-exposed dermal thickness was decreased in comparison with the UVA-protected 3D skin model, recovered with GERg3 treatment. GERg3 had antiphotoaging effects in UV- or IR-exposed human dermal fibroblasts and normal human 3D skin model. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Top3-Rmi1 dissolve Rad51-mediated D-loops by a topoisomerase-based mechanism

PubMed Central

Fasching, Clare L.; Cejka, Petr; Kowalczykowski, Stephen C.; Heyer, Wolf-Dietrich

2015-01-01

Summary The displacement loop (D-loop) is the DNA strand invasion product formed during homologous recombination. Disruption of nascent D-loops represents a mechanism of anti-recombination. During Synthesis-Dependent Strand Annealing D-loop disruption after extension of the invading strand is an integral step of the pathway and ensures a non-crossover outcome. The proteins implicated in D-loop disruption are DNA motor proteins/helicases acting by migrating DNA junctions. Here we report an unanticipated mechanism of D-loop dissolution mediated by DNA topoisomerase 3 (Top3) and dependent on its catalytic activity. D-loop dissolution catalyzed by yeast Top3 is highly specific for yeast Rad51/Rad54-mediated D-loops, whereas protein-free D-loops or D-loop mediated by bacterial RecA protein or human RAD51/RAD54 resist dissolution. Also the human Topoisomerase IIIα-RMI1–RMI2 complex is capable of dissolving D-loops. Consistent with genetic data, we suggest that the extreme growth defect and hyper-recombination phenotype of Top3-deficient yeast cells is in part a result of unprocessed D-loops. PMID:25699708

Separate Perceptual and Neural Processing of Velocity- and Disparity-Based 3D Motion Signals

PubMed Central

Czuba, Thaddeus B.; Cormack, Lawrence K.; Huk, Alexander C.

2016-01-01

Although the visual system uses both velocity- and disparity-based binocular information for computing 3D motion, it is unknown whether (and how) these two signals interact. We found that these two binocular signals are processed distinctly at the levels of both cortical activity in human MT and perception. In human MT, adaptation to both velocity-based and disparity-based 3D motions demonstrated direction-selective neuroimaging responses. However, when adaptation to one cue was probed using the other cue, there was no evidence of interaction between them (i.e., there was no “cross-cue” adaptation). Analogous psychophysical measurements yielded correspondingly weak cross-cue motion aftereffects (MAEs) in the face of very strong within-cue adaptation. In a direct test of perceptual independence, adapting to opposite 3D directions generated by different binocular cues resulted in simultaneous, superimposed, opposite-direction MAEs. These findings suggest that velocity- and disparity-based 3D motion signals may both flow through area MT but constitute distinct signals and pathways. SIGNIFICANCE STATEMENT Recent human neuroimaging and monkey electrophysiology have revealed 3D motion selectivity in area MT, which is driven by both velocity-based and disparity-based 3D motion signals. However, to elucidate the neural mechanisms by which the brain extracts 3D motion given these binocular signals, it is essential to understand how—or indeed if—these two binocular cues interact. We show that velocity-based and disparity-based signals are mostly separate at the levels of both fMRI responses in area MT and perception. Our findings suggest that the two binocular cues for 3D motion might be processed by separate specialized mechanisms. PMID:27798134

Separate Perceptual and Neural Processing of Velocity- and Disparity-Based 3D Motion Signals.

PubMed

Joo, Sung Jun; Czuba, Thaddeus B; Cormack, Lawrence K; Huk, Alexander C

2016-10-19

Although the visual system uses both velocity- and disparity-based binocular information for computing 3D motion, it is unknown whether (and how) these two signals interact. We found that these two binocular signals are processed distinctly at the levels of both cortical activity in human MT and perception. In human MT, adaptation to both velocity-based and disparity-based 3D motions demonstrated direction-selective neuroimaging responses. However, when adaptation to one cue was probed using the other cue, there was no evidence of interaction between them (i.e., there was no "cross-cue" adaptation). Analogous psychophysical measurements yielded correspondingly weak cross-cue motion aftereffects (MAEs) in the face of very strong within-cue adaptation. In a direct test of perceptual independence, adapting to opposite 3D directions generated by different binocular cues resulted in simultaneous, superimposed, opposite-direction MAEs. These findings suggest that velocity- and disparity-based 3D motion signals may both flow through area MT but constitute distinct signals and pathways. Recent human neuroimaging and monkey electrophysiology have revealed 3D motion selectivity in area MT, which is driven by both velocity-based and disparity-based 3D motion signals. However, to elucidate the neural mechanisms by which the brain extracts 3D motion given these binocular signals, it is essential to understand how-or indeed if-these two binocular cues interact. We show that velocity-based and disparity-based signals are mostly separate at the levels of both fMRI responses in area MT and perception. Our findings suggest that the two binocular cues for 3D motion might be processed by separate specialized mechanisms. Copyright © 2016 the authors 0270-6474/16/3610791-12$15.00/0.

Real-time 3D visualization of volumetric video motion sensor data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Carlson, J.; Stansfield, S.; Shawver, D.

1996-11-01

This paper addresses the problem of improving detection, assessment, and response capabilities of security systems. Our approach combines two state-of-the-art technologies: volumetric video motion detection (VVMD) and virtual reality (VR). This work capitalizes on the ability of VVMD technology to provide three-dimensional (3D) information about the position, shape, and size of intruders within a protected volume. The 3D information is obtained by fusing motion detection data from multiple video sensors. The second component involves the application of VR technology to display information relating to the sensors and the sensor environment. VR technology enables an operator, or security guard, to bemore » immersed in a 3D graphical representation of the remote site. VVMD data is transmitted from the remote site via ordinary telephone lines. There are several benefits to displaying VVMD information in this way. Because the VVMD system provides 3D information and because the sensor environment is a physical 3D space, it seems natural to display this information in 3D. Also, the 3D graphical representation depicts essential details within and around the protected volume in a natural way for human perception. Sensor information can also be more easily interpreted when the operator can `move` through the virtual environment and explore the relationships between the sensor data, objects and other visual cues present in the virtual environment. By exploiting the powerful ability of humans to understand and interpret 3D information, we expect to improve the means for visualizing and interpreting sensor information, allow a human operator to assess a potential threat more quickly and accurately, and enable a more effective response. This paper will detail both the VVMD and VR technologies and will discuss a prototype system based upon their integration.« less

3D Cones Acquisition of Human Extremity Imaging Using a 1.5T Superconducting Magnet and an Unshielded Gradient Coil Set.

PubMed

Setoi, Ayana; Kose, Katsumi

2018-05-16

We developed ultrashort echo-time (UTE) imaging sequences with 3D Cones trajectories for a home-built compact MRI system using a 1.5T superconducting magnet and an unshielded gradient coil set. We achieved less than 7 min imaging time and obtained clear in vivo images of a human forearm with a TE of 0.4 ms. We concluded that UTE imaging using 3D Cones acquisition was successfully implemented in our 1.5T MRI system.

Organotypic culture in three dimensions prevents radiation-induced transformation in human lung epithelial cells

NASA Astrophysics Data System (ADS)

El-Ashmawy, Mariam; Coquelin, Melissa; Luitel, Krishna; Batten, Kimberly; Shay, Jerry W.

2016-08-01

The effects of radiation in two-dimensional (2D) cell culture conditions may not recapitulate tissue responses as modeled in three-dimensional (3D) organotypic culture. In this study, we determined if the frequency of radiation-induced transformation and cancer progression differed in 3D compared to 2D culture. Telomerase immortalized human bronchial epithelial cells (HBECs) with shTP53 and mutant KRas expression were exposed to various types of radiation (gamma, +H, 56Fe) in either 2D or 3D culture. After irradiation, 3D structures were dissociated and passaged as a monolayer followed by measurement of transformation, cell growth and expression analysis. Cells irradiated in 3D produced significantly fewer and smaller colonies in soft agar than their 2D-irradiated counterparts (gamma P = 0.0004 +H P = 0.049 56Fe P < 0.0001). The cell culture conditions did not affect cell killing, the ability of cells to survive in a colony formation assay, and proliferation rates after radiation—implying there was no selection against cells in or dissociated from 3D conditions. However, DNA damage repair and apoptosis markers were increased in 2D cells compared to 3D cells after radiation. Ideally, expanding the utility of 3D culture will allow for a better understanding of the biological consequences of radiation exposure.

Vitamin D alters genes involved in follicular development and steroidogenesis in human cumulus granulosa cells.

PubMed

Merhi, Zaher; Doswell, Angela; Krebs, Kendall; Cipolla, Marilyn

2014-06-01

Vitamin D deficiency is common among reproductive-aged women and has a role in female reproduction. This study evaluated the role of 1,25-dihydroxyvitamin D3 (vit D3) in ovarian follicular development and steroidogenesis by using a human granulosa cell (GC) model. Fifty-four women who underwent in vitro fertilization were enrolled. Follicular fluid (FF) and mural and cumulus GCs were collected from small and large follicles. In separate experiments, primary cumulus GCs were cultured with or without vit D3 followed by RT-PCR for mRNA expression levels. The effect of recombinant anti-Mullerian hormone (AMH) on nuclear localization of phospho-Smad 1/5/8 was evaluated in the presence or absence of vit D3 by using immunofluorescence. 25-Hydroxyvitamin D levels in FF as well as cell culture media AMH, progesterone, and estradiol (E2) concentrations were determined by ELISA and RIA. The following were measured: 1) mRNA expression levels; 2) 3β-hydroxysteroid dehydrogenase (3β-HSD) enzyme activity; 3) FSH-induced aromatase mRNA and E2 production; and 4) nuclear localization of phospho-Smad 1/5/8. In a multivariate analysis, 25 OH-D levels in FF negatively correlated with AMH and AMH receptor (AMHR)-II mRNA levels in cumulus GCs of small follicles. Compared with women with replete 25-hydroxyvitamin D levels in FF, those with insufficient/deficient levels had a 2-fold increase in AMHR-II mRNA levels in cumulus GCs of small follicles (P = .02). Treatment with vit D3 caused a decrease in AMHR-II and FSH receptor mRNA but an increase in 3-βHSD mRNA levels compared with control (P < .05). Vit D3 enhanced 3-βHSD enzyme activity as assessed by increasing progesterone release; however, vit D3 did not affect FSH-induced aromatase mRNA and E2 production, but it decreased the phosphorylation of Smad 1/5/8 and its nuclear localization. These data suggest that vit D3 alters AMH signaling and steroidogenesis in human cumulus GCs, possibly reflecting a state of GC luteinization potentiation.

Role of the National Institute of Standards and Technology (NIST) in Support of the Vitamin D Initiative of the National Institutes of Health, Office of Dietary Supplements.

PubMed

Wise, Stephen A; Tai, Susan S-C; Burdette, Carolyn Q; Camara, Johanna E; Bedner, Mary; Lippa, Katrice A; Nelson, Michael A; Nalin, Federica; Phinney, Karen W; Sander, Lane C; Betz, Joseph M; Sempos, Christopher T; Coates, Paul M

2017-09-01

Since 2005, the National Institute of Standards and Technology (NIST) has collaborated with the National Institutes of Health (NIH), Office of Dietary Supplements (ODS) to improve the quality of measurements related to human nutritional markers of vitamin D status. In support of the NIH-ODS Vitamin D Initiative, including the Vitamin D Standardization Program (VDSP), NIST efforts have focused on (1) development of validated analytical methods, including reference measurement procedures (RMPs); (2) development of Standard Reference Materials (SRMs); (3) value assignment of critical study samples using NIST RMPs; and (4) development and coordination of laboratory measurement QA programs. As a result of this collaboration, NIST has developed RMPs for 25-hydroxyvitamin D2 [25(OH)D2], 25(OH)D3, and 24R,25-dihydroxyvitamin D3 [24R,25(OH)2D3]; disseminated serum-based SRMs with values assigned for 25(OH)D2, 25(OH)D3, 3-epi-25(OH)D3, and 24R,25(OH)2D3; assigned values for critical samples for VDSP studies, including an extensive interlaboratory comparison and reference material commutability study; provided an accuracy basis for the Vitamin D External Quality Assurance Scheme; coordinated the first accuracy-based measurement QA program for the determination of 25(OH)D2, 25(OH)D3, and 3-epi-25(OH)D3 in human serum/plasma; and developed methods and SRMs for the determination of vitamin D and 25(OH)D in food and supplement matrix SRMs. The details of these activities and their benefit and impact to the NIH-ODS Vitamin D Initiative are described.

42 CFR 51d.3 - Who is eligible for an award under this subpart?

Code of Federal Regulations, 2012 CFR

2012-10-01

... 42 Public Health 1 2012-10-01 2012-10-01 false Who is eligible for an award under this subpart? 51d.3 Section 51d.3 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS MENTAL HEALTH AND SUBSTANCE ABUSE EMERGENCY RESPONSE PROCEDURES § 51d.3 Who is eligible for an...

42 CFR 51d.3 - Who is eligible for an award under this subpart?

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 1 2011-10-01 2011-10-01 false Who is eligible for an award under this subpart? 51d.3 Section 51d.3 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS MENTAL HEALTH AND SUBSTANCE ABUSE EMERGENCY RESPONSE PROCEDURES § 51d.3 Who is eligible for an...

42 CFR 51d.3 - Who is eligible for an award under this subpart?

Code of Federal Regulations, 2014 CFR

2014-10-01

... 42 Public Health 1 2014-10-01 2014-10-01 false Who is eligible for an award under this subpart? 51d.3 Section 51d.3 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS MENTAL HEALTH AND SUBSTANCE ABUSE EMERGENCY RESPONSE PROCEDURES § 51d.3 Who is eligible for an...

42 CFR 51d.3 - Who is eligible for an award under this subpart?

Code of Federal Regulations, 2013 CFR

2013-10-01

... 42 Public Health 1 2013-10-01 2013-10-01 false Who is eligible for an award under this subpart? 51d.3 Section 51d.3 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS MENTAL HEALTH AND SUBSTANCE ABUSE EMERGENCY RESPONSE PROCEDURES § 51d.3 Who is eligible for an...

42 CFR 51d.3 - Who is eligible for an award under this subpart?

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 1 2010-10-01 2010-10-01 false Who is eligible for an award under this subpart? 51d.3 Section 51d.3 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS MENTAL HEALTH AND SUBSTANCE ABUSE EMERGENCY RESPONSE PROCEDURES § 51d.3 Who is eligible for an...

Single shot laser speckle based 3D acquisition system for medical applications

NASA Astrophysics Data System (ADS)

Khan, Danish; Shirazi, Muhammad Ayaz; Kim, Min Young

2018-06-01

The state of the art techniques used by medical practitioners to extract the three-dimensional (3D) geometry of different body parts requires a series of images/frames such as laser line profiling or structured light scanning. Movement of the patients during scanning process often leads to inaccurate measurements due to sequential image acquisition. Single shot structured techniques are robust to motion but the prevalent challenges in single shot structured light methods are the low density and algorithm complexity. In this research, a single shot 3D measurement system is presented that extracts the 3D point cloud of human skin by projecting a laser speckle pattern using a single pair of images captured by two synchronized cameras. In contrast to conventional laser speckle 3D measurement systems that realize stereo correspondence by digital correlation of projected speckle patterns, the proposed system employs KLT tracking method to locate the corresponding points. The 3D point cloud contains no outliers and sufficient quality of 3D reconstruction is achieved. The 3D shape acquisition of human body parts validates the potential application of the proposed system in the medical industry.

M-BAND analysis of chromosome aberration induced by Fe-ions in human epithelial cells cultured in 3-dimensional matrices

NASA Astrophysics Data System (ADS)

Hada, Megumi; Cucinotta, Francis A.; Wu, Honglu

Energetic heavy ions pose a great health risk to astronauts in extended ISS and future lunar and Mars missions. High-LET heavy ions are particularly effective in causing various biological effects, including cell inactivation, genetic mutations, cataracts and cancer induction. Most of these biological endpoints are closely related to chromosomal damage, which can be utilized as a biomarker for radiation insults. Previously, we had studied lowand high-LET radiationinduced chromosome aberrations in human epithelial cells cultured in 2-dimension (2D) using the multicolor banding fluorescence in situ hybridization (mBAND) technique. However, it has been realized that the biological response to radiation insult in a 2D cellular environment in vitro can differ significantly from the response in 3-dimension (3D) or at the actual tissue level. In this study, we cultured human epithelial cells in 3D to provide a more suitable model for human tissue. Human mammary epithelial cells (CH184B5F5/M10) were grown in Matrigel to form 3D structures, and exposed to Fe-ions at NASA Space Radiation Laboratory (NSRL) at the Brookhaven National Laboratory or 137 Cs-gamma radiation source at the University of Texas MD Anderson Cancer Center. After exposure, cells were allowed to repair for 16hr before dissociation and subcultured at low density in 2D. G2 and metaphase chromosomes in the first cell cycle were collected using a chemical-induced premature chromosome condensation (PCC) technique, and chromosome aberrations were analyzed using mBAND technique. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of interchromosomal aberrations (translocation to unpainted chromosomes) and intrachromosomal aberrations (inversions and deletions within a single painted chromosome). Our data indicate a significant difference of the chromosome aberration yield between 2D and 3D cell cultures for gamma exposures, but not for Fe ion exposures. Therefore, the RBE for chromosome aberrations obtained in a 2D model may not represent accurately the RBE for tissues.

High- and low-LET Radiation-induced Chromosome Aberrations in Human Epithelial Cells Cultured in 3-dimensional Matrices

NASA Technical Reports Server (NTRS)

Hada, M.; George K.; Cucinotta, F. A.; Wu, H.

2008-01-01

Energetic heavy ions pose a great health risk to astronauts who participate in extended ISS missions and will be an even greater concern for future manned lunar and Mars missions. High-LET heavy ions are particularly effective in causing various biological effects, including cell inactivation, genetic mutations, cataracts and cancer induction. Most of these biological endpoints are closely related to chromosomal damage, which can be utilized as a biomarker for radiation insults. Previously, we had studied low- and high-LET radiation-induced chromosome aberrations in human epithelial cells cultured in 2-dimension (2D) using the multicolor banding fluorescence in situ hybridization (mBAND) technique. However, it has been realized that the biological response to radiation insult in a 2D in vitro cellular environment can differ significantly from the response in 3-dimension (3D) or at the actual tissue level. In this study, we cultured human epithelial cells in 3D to provide a more suitable model for human tissue. Human mammary epithelial cells (CH184B5F5/M10) were grown in Matrigel to form 3D structures, and exposed to Fe-ions at NASA Space Radiation Laboratory (NSRL) at the Brookhaven National Laboratory or 137Cs-gamma radiation source at the University of Texas MD Anderson Cancer Center. After exposure, cells were allowed to repair for 16hr before dissociation and subcultured at low density in 2D. G2 and metaphase chromosomes in the first cell cycle were collected in the first cell cycle after irradiation using a chemical-induced premature chromosome condensation (PCC) technique, and chromosome aberrations were analyzed using mBAND technique. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of interchromosomal aberrations (translocation to unpainted chromosomes) and intrachromosomal aberrations (inversions and deletions within a single painted chromosome). Our data indicate a significant difference in the chromosome aberration yield between 2D and 3D cell cultures after gamma exposures, but not after Fe ion exposures. Therefore, the Relative Biological Effect (RBE) for induction of chromosome aberrations obtained in a 2D model may not accurately represent RBE values obtained for tissue exposure.

M-BAND Analysis of Chromosome Aberration Induced by Fe-Ions in Human Epithelial Cells Cultured in 3-Dimensional Matrices

NASA Technical Reports Server (NTRS)

Hada, M.; Cucinotta, F. A.; Wu, H.

2008-01-01

Energetic heavy ions pose a great health risk to astronauts in extended ISS and future lunar and Mars missions. High-LET heavy ions are particularly effective in causing various biological effects, including cell inactivation, genetic mutations, cataracts and cancer induction. Most of these biological endpoints are closely related to chromosomal damage, which can be utilized as a biomarker for radiation insults. Previously, we had studied low- and high-LET radiation-induced chromosome aberrations in human epithelial cells cultured in 2-dimension (2D) using the multicolor banding fluorescence in situ hybridization (mBAND) technique. However, it has been realized that the biological response to radiation insult in a 2D cellular environment in vitro can differ significantly from the response in 3-dimension (3D) or at the actual tissue level. In this study, we cultured human epithelial cells in 3D to provide a more suitable model for human tissue. Human mammary epithelia cells (CH184B5F5/M10) were grown in Matrigel to form 3D structures, and exposed to Fe-ions at NASA Space Radiation Laboratory (NSRL) at the Brookhaven National Laboratory or 137Cs-gamma radiation source at the University of Texas MD Anderson Cancer Center. After exposure, cells were allowed to repair for 16hr before dissociation and subcultued at low density in 2D. G2 and metaphase chromosomes in the first cell cycle were collected using a chemical-induced premature chromosome condensation (PCC) technique, and chromosome aberrations were analyzed using mBAND technique. With this technique, individually painted chromosomal bands on one chromosome allowed the identification of interchromosomal aberrations (translocation to unpainted chromosomes) and intrachromosomal aberrations (inversions and deletions within a single painted chromosome). Our data indicate a significant difference of the chromosome aberration yield between 2D and 3D cell cultures for gamma exposures, but not for Fe ion exposures. Therefore, the RBE for chromosome aberrations obtained in a 2D model may not represent accurately the RBE for tissues.

Non-invasive 3D geometry extraction and robotic modeling of a Sea lion foreflipper

NASA Astrophysics Data System (ADS)

Patel, R. K.; Leftwich, M. C.; Friedman, C.

2016-02-01

California Sea Lions are very agile swimmers and unlike many marine animals, they use their fore flipper rather than their hind flipper undulations to generate high thrust values. To date there exist limited amount of qualitative studies for sea lions swimming that show the flippers are used for thrust, stability, and control during swimming motions. Quantitative studies mainly measured drag used for cost of transport, and analyzed banked turn performance. Recently, the kinematics of a California sea lion flipper during the thrust phase was extracted using video tracking in two dimensions. This work extends the tracking ability to three dimensions using a non-invasive Direct Linear Transformation (DLT) technique employed on non-research sea lions at the Smithsonian National Zoological Park. The flippers are therefore marker-less and tracking is carried out manually in post processing after capturing complete dorsal-ventral flipper motions. Two cameras are used (3840 × 2160 pixels resolution) and calibrated in space using a calibration target inserted into the sea lion habitat. They are synchronized in time using a simple light flash. The fluid flow and forces generated by a sea lion clap is also being explored. Recently, a sea lion flipper from a deceased subject was externally scanned in high detail for fluid dynamics research. The flipper's geometry is being used in this work to design and build an articulate flipper model that is approximately 60% of the full size span. The model is actuated by servo motors and is designed to mimic a sea lion flipper clap motion based on the previously extracted kinematics from above. The model incorporates three axles, simulating the movements of the sea lion's elbow, wrist, and knuckles. The flipper tip speed is designed to match typical Reynolds numbers for the full-scale flipper for an acceleration from rest maneuver. The model will be tested in a water flume to obtain the forces during the thrust production phase of the flipper motion.

1α,25(OH)2D3 differentially regulates miRNA expression in human bladder cancer cells

PubMed Central

Ma, Yingyu; Hu, Qiang; Luo, Wei; Pratt, Rachel N.; Glenn, Sean T.; Liu, Song; Trump, Donald L.; Johnson, Candace S.

2014-01-01

Bladder cancer is the fourth most commonly diagnosed cancer in men and eighth leading cause of cancer-related death in the US. Epidemiological and experimental studies strongly suggest a role for 1α,25(OH)2D3 in cancer prevention and treatment. The antitumor activities of 1α,25(OH)2D3 are mediated by the induction of cell cycle arrest, apoptosis, differentiation and the inhibition of angiogenesis and metastasis. MiRNAs play important regulatory roles in cancer development and progression. However, the role of 1α,25(OH)2D3 in the regulation of miRNA expression and the potential impact in bladder cancer has not been investigated. Therefore, we studied 1α,25(OH)2D3-regulated miRNA expression profiles in human bladder cancer cell line 253J and the highly tumorigenic and metastatic derivative line 253J-BV by miRNA qPCR panels. 253 J and 253J-BV cells express endogenous vitamin D receptor (VDR) which can be further induced by 1α,25(OH)2D3. VDR target gene 24-hydroxylase was induced by 1α,25(OH)2D3 in both cell lines, indicating functional 1α,25(OH)2D3 signaling. The miRNA qPCR panel assay results showed that 253J and 253J-BV cells have distinct miRNA expression profiles. Further, 1α,25(OH)2D3 differentially regulated miRNA expression profiles in 253J and 253 J-BV cells in a dynamic manner. Pathway analysis of the miRNA target genes revealed distinct patterns of contribution to the molecular functions and biological processes in the two cell lines. In conclusion, 1α,25(OH)2D3 differentially regulates the expression of miRNAs, which may contribute to distinct biological functions, in human bladder 253J and 253J-BV cells. PMID:25263658

Perfusion Stirred-Tank Bioreactors for 3D Differentiation of Human Neural Stem Cells.

PubMed

Simão, Daniel; Arez, Francisca; Terasso, Ana P; Pinto, Catarina; Sousa, Marcos F Q; Brito, Catarina; Alves, Paula M

2016-01-01

Therapeutic breakthroughs in neurological disorders have been hampered by the lack of accurate central nervous system (CNS) models. The development of these models allows the study of the disease onset/progression mechanisms and the preclinical evaluation of new therapeutics. This has traditionally relied on genetically engineered animal models that often diverge considerably from the human phenotype (developmental, anatomic, and physiological) and 2D in vitro cell models, which fail to recapitulate the characteristics of the target tissue (cell-cell and cell-matrix interactions, cell polarity, etc.). Recapitulation of CNS phenotypic and functional features in vitro requires the implementation of advanced culture strategies, such as 3D culture systems, which enable to mimic the in vivo structural and molecular complexity. Models based on differentiation of human neural stem cells (hNSC) in 3D cultures have great potential as complementary tools in preclinical research, bridging the gap between human clinical studies and animal models. The development of robust and scalable processes for the 3D differentiation of hNSC can improve the accuracy of early stage development in preclinical research. In this context, the use of software-controlled stirred-tank bioreactors (STB) provides an efficient technological platform for hNSC aggregation and differentiation. This system enables to monitor and control important physicochemical parameters for hNSC culture, such as dissolved oxygen. Importantly, the adoption of a perfusion operation mode allows a stable flow of nutrients and differentiation/neurotrophic factors, while clearing the toxic by-products. This contributes to a setting closer to the physiological, by mimicking the in vivo microenvironment. In this chapter, we address the technical requirements and procedures for the implementation of 3D differentiation strategies of hNSC, by operating STB under perfusion mode for long-term cultures. This strategy is suitable for the generation of human 3D neural in vitro models, which can be used to feed high-throughput screening platforms, contributing to expand the available in vitro tools for drug screening and toxicological studies.

Suitability of markerless EPID tracking for tumor position verification in gated radiotherapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Serpa, Marco; University Clinic for Radiotherapy and Radio-Oncology, Landeskrankenhaus Salzburg, Paracelsus Medical University Clinics, 5020 Salzburg; Department of Physics and Astronomy, University of Canterbury, Christchurch 8140

2014-03-15

Purpose: To maximize the benefits of respiratory gated radiotherapy (RGRT) of lung tumors real-time verification of the tumor position is required. This work investigates the feasibility of markerless tracking of lung tumors during beam-on time in electronic portal imaging device (EPID) images of the MV therapeutic beam. Methods: EPID movies were acquired at ∼2 fps for seven lung cancer patients with tumor peak-to-peak motion ranges between 7.8 and 17.9 mm (mean: 13.7 mm) undergoing stereotactic body radiotherapy. The external breathing motion of the abdomen was synchronously measured. Both datasets were retrospectively analyzed inPortalTrack, an in-house developed tracking software. The authorsmore » define a three-step procedure to run the simulations: (1) gating window definition, (2) gated-beam delivery simulation, and (3) tumor tracking. First, an amplitude threshold level was set on the external signal, defining the onset of beam-on/-off signals. This information was then mapped onto a sequence of EPID images to generate stamps of beam-on/-hold periods throughout the EPID movies in PortalTrack, by obscuring the frames corresponding to beam-off times. Last, tumor motion in the superior-inferior direction was determined on portal images by the tracking algorithm during beam-on time. The residual motion inside the gating window as well as target coverage (TC) and the marginal target displacement (MTD) were used as measures to quantify tumor position variability. Results: Tumor position monitoring and estimation from beam's-eye-view images during RGRT was possible in 67% of the analyzed beams. For a reference gating window of 5 mm, deviations ranging from 2% to 86% (35% on average) were recorded between the reference and measured residual motion. TC (range: 62%–93%; mean: 77%) losses were correlated with false positives incidence rates resulting mostly from intra-/inter-beam baseline drifts, as well as sudden cycle-to-cycle fluctuations in exhale positions. Both phenomena can lead to considerable deviations (with MTD values up to a maximum of 7.8 mm) from the intended tumor position, and in turn may result in a marginal miss. The difference between tumor traces determined within the gating window against ground truth trajectory maps was 1.1 ± 0.7 mm on average (range: 0.4–2.3 mm). Conclusions: In this retrospective analysis of motion data, it is demonstrated that the system is capable of determining tumor positions in the plane perpendicular to the beam direction without the aid of fiducial markers, and may hence be suitable as an online verification tool in RGRT. It may be possible to use the tracking information to enable on-the-fly corrections to intra-/inter-beam variations by adapting the gating window by means of a robotic couch.« less

The potential failure risk of the cone-beam computed tomography-based planning target volume margin definition for prostate image-guided radiotherapy based on a prospective single-institutional hybrid analysis.

PubMed

Hirose, Katsumi; Sato, Mariko; Hatayama, Yoshiomi; Kawaguchi, Hideo; Komai, Fumio; Sohma, Makoto; Obara, Hideki; Suzuki, Masashi; Tanaka, Mitsuki; Fujioka, Ichitaro; Ichise, Koji; Takai, Yoshihiro; Aoki, Masahiko

2018-06-07

The purpose of this study was to evaluate the impact of markerless on-board kilovoltage (kV) cone-beam computed tomography (CBCT)-based positioning uncertainty on determination of the planning target volume (PTV) margin by comparison with kV on-board imaging (OBI) with gold fiducial markers (FMs), and to validate a methodology for the evaluation of PTV margins for markerless kV-CBCT in prostate image-guided radiotherapy (IGRT). A total of 1177 pre- and 1177 post-treatment kV-OBI and 1177 pre- and 206 post-treatment kV-CBCT images were analyzed in 25 patients who received prostate IGRT with daily localization by implanted FMs. Intrafractional motion of the prostate was evaluated between each pre- and post-treatment image with these two different techniques. The differences in prostate deviations and intrafractional motions between matching by FM in kV-OBI (OBI-FM) and matching by soft tissues in kV-CBCT (CBCT-ST) were compared by Bland-Altman limits of agreement. Compensated PTV margins were determined and compensated by references. Mean differences between OBI-FM and CBCT-ST in the anterior to posterior (AP), superior to inferior (SI), and left to right (LR) directions were - 0.43 ± 1.45, - 0.09 ± 1.65, and - 0.12 ± 0.80 mm, respectively, with R 2  = 0.85, 0.88, and 0.83, respectively. Intrafractional motions obtained from CBCT-ST were 0.00 ± 1.46, 0.02 ± 1.49, and 0.15 ± 0.64 mm, respectively, which were smaller than the results from OBI-FM, with 0.43 ± 1.90, 0.12 ± 1.98, and 0.26 ± 0.80 mm, respectively, with R 2  = 0.42, 0.33, and 0.16, respectively. Bland-Altman analysis showed a significant proportional bias. PTV margins of 1.5 mm, 1.4 mm, and 0.9 mm for CBCT-ST were calculated from the values of CBCT-ST, which were also smaller than the values of 3.15 mm, 3.66 mm, and 1.60 mm from OBI-FM. The practical PTV margin for CBCT-ST was compensated with the values from OBI-FM as 4.1 mm, 4.8 mm, and 2.2 mm. PTV margins calculated from CBCT-ST might be underestimated compared to the true PTV margins. To determine a reliable CBCT-ST-based PTV margin, at least the systemic error Σ and the random error σ for on-line matching errors need to be investigated by supportive preliminary FM evaluation at least once.

Young children’s ability to use 2-dimensional and 3-dimensional symbols to show placements of body touches and hidden objects

PubMed Central

Lytle, Nicole; London, Kamala; Bruck, Maggie

2015-01-01

In two experiments, we investigated 3- to 5-year-old children’s ability to use dolls and human figure drawings as symbols to map body touches. In Experiment 1 stickers were placed on different locations of children’s bodies, and they were asked to indicate the location of the sticker using three different symbols: a doll, a human figure drawing, and the adult researcher. Performance on the tasks increased with age, but many 5-year-olds did not attain perfect performance. Surprisingly, younger children made more errors on the 2D human figure drawing task compared to the 3D doll and adult tasks. In Experiment 2, we compared children’s ability to use 3D and 2D symbols to indicate body touch as well as to guide their search for a hidden object. We replicated the findings of Experiment 1 for the body touch task: for younger children, 3D symbols were easier to use than 2D symbols. However, the reverse pattern was found for the object locations task with children showing superior performance using 2D drawings over 3D models. Though children showed developmental improvements in using dolls and drawings to show where they were touched, less than two-thirds of the 5-year-olds performed perfectly on the touch tasks. Developmental as well as forensic implications of these results are discussed. PMID:25781003

The Extraction of 3D Shape from Texture and Shading in the Human Brain

PubMed Central

Georgieva, Svetlana S.; Todd, James T.; Peeters, Ronald

2008-01-01

We used functional magnetic resonance imaging to investigate the human cortical areas involved in processing 3-dimensional (3D) shape from texture (SfT) and shading. The stimuli included monocular images of randomly shaped 3D surfaces and a wide variety of 2-dimensional (2D) controls. The results of both passive and active experiments reveal that the extraction of 3D SfT involves the bilateral caudal inferior temporal gyrus (caudal ITG), lateral occipital sulcus (LOS) and several bilateral sites along the intraparietal sulcus. These areas are largely consistent with those involved in the processing of 3D shape from motion and stereo. The experiments also demonstrate, however, that the analysis of 3D shape from shading is primarily restricted to the caudal ITG areas. Additional results from psychophysical experiments reveal that this difference in neuronal substrate cannot be explained by a difference in strength between the 2 cues. These results underscore the importance of the posterior part of the lateral occipital complex for the extraction of visual 3D shape information from all depth cues, and they suggest strongly that the importance of shading is diminished relative to other cues for the analysis of 3D shape in parietal regions. PMID:18281304

Comparisons of human amniotic mesenchymal stem cell viability in FDA-approved collagen-based scaffolds: Implications for engineered diaphragmatic replacement.

PubMed

Shieh, Hester F; Graham, Christopher D; Brazzo, Joseph A; Zurakowski, David; Fauza, Dario O

2017-06-01

We sought to examine amniotic fluid mesenchymal stem cell (afMSC) viability within two FDA-approved collagen-based scaffolds, as a prerequisite to clinical translation of afMSC-based engineered diaphragmatic repair. Human afMSCs were seeded in a human-derived collagen hydrogel and in a bovine-derived collagen sheet at 3 matching densities. Cell viability was analyzed at 1, 3, and 5days using an ATP-based 3D bioluminescence assay. Statistical comparisons were by ANOVA (P<0.05). There was a highly significant 3-way interaction between scaffold type, seeding density, and time in 3D culture as determinants of cell viability, clearly favoring the human hydrogel (P<0.001). In both scaffolds, cell viability was highest at the highest seeding density of 150,000 cells/mL. Time in 3D culture impacted cell viability at the optimal seeding density in the human hydrogel, with the highest levels on days 1 (P<0.001) and 5 (P=0.05) with no significant effect in the bovine sheet (P=0.39-0.96). Among clinically-approved cell delivery vehicles, mesenchymal stem cell viability is significantly enhanced in a collagen hydrogel when compared with a collagen sheet. Cell viability can be further optimized by seeding density and time in 3D culture. These data further support the regulatory viability of clinical trials of engineered diaphragmatic repair. N/A (animal and laboratory study). Copyright © 2017 Elsevier Inc. All rights reserved.

Experimental design based 3-D QSAR analysis of steroid-protein interactions: Application to human CBG complexes

NASA Astrophysics Data System (ADS)

Norinder, Ulf

1990-12-01

An experimental design based 3-D QSAR analysis using a combination of principal component and PLS analysis is presented and applied to human corticosteroid-binding globulin complexes. The predictive capability of the created model is good. The technique can also be used as guidance when selecting new compounds to be investigated.

A Virtual Campus Based on Human Factor Engineering

ERIC Educational Resources Information Center

Yang, Yuting; Kang, Houliang

2014-01-01

Three Dimensional or 3D virtual reality has become increasingly popular in many areas, especially in building a digital campus. This paper introduces a virtual campus, which is based on a 3D model of The Tourism and Culture College of Yunnan University (TCYU). Production of the virtual campus was aided by Human Factor and Ergonomics (HF&E), an…

Technical Note: Guidelines for the digital computation of 2D and 3D enamel thickness in hominoid teeth.

PubMed

Benazzi, Stefano; Panetta, Daniele; Fornai, Cinzia; Toussaint, Michel; Gruppioni, Giorgio; Hublin, Jean-Jacques

2014-02-01

The study of enamel thickness has received considerable attention in regard to the taxonomic, phylogenetic and dietary assessment of human and non-human primates. Recent developments based on two-dimensional (2D) and three-dimensional (3D) digital techniques have facilitated accurate analyses, preserving the original object from invasive procedures. Various digital protocols have been proposed. These include several procedures based on manual handling of the virtual models and technical shortcomings, which prevent other scholars from confidently reproducing the entire digital protocol. There is a compelling need for standard, reproducible, and well-tailored protocols for the digital analysis of 2D and 3D dental enamel thickness. In this contribution we provide essential guidelines for the digital computation of 2D and 3D enamel thickness in hominoid molars, premolars, canines and incisors. We modify previous techniques suggested for 2D analysis and we develop a new approach for 3D analysis that can also be applied to premolars and anterior teeth. For each tooth class, the cervical line should be considered as the fundamental morphological feature both to isolate the crown from the root (for 3D analysis) and to define the direction of the cross-sections (for 2D analysis). Copyright © 2013 Wiley Periodicals, Inc.

Melanin Transfer in Human 3D Skin Equivalents Generated Exclusively from Induced Pluripotent Stem Cells

PubMed Central

Gledhill, Karl; Guo, Zongyou; Umegaki-Arao, Noriko; Higgins, Claire A.; Itoh, Munenari; Christiano, Angela M.

2015-01-01

The current utility of 3D skin equivalents is limited by the fact that existing models fail to recapitulate the cellular complexity of human skin. They often contain few cell types and no appendages, in part because many cells found in the skin are difficult to isolate from intact tissue and cannot be expanded in culture. Induced pluripotent stem cells (iPSCs) present an avenue by which we can overcome this issue due to their ability to be differentiated into multiple cell types in the body and their unlimited growth potential. We previously reported generation of the first human 3D skin equivalents from iPSC-derived fibroblasts and iPSC-derived keratinocytes, demonstrating that iPSCs can provide a foundation for modeling a complex human organ such as skin. Here, we have increased the complexity of this model by including additional iPSC-derived melanocytes. Epidermal melanocytes, which are largely responsible for skin pigmentation, represent the second most numerous cell type found in normal human epidermis and as such represent a logical next addition. We report efficient melanin production from iPSC-derived melanocytes and transfer within an entirely iPSC-derived epidermal-melanin unit and generation of the first functional human 3D skin equivalents made from iPSC-derived fibroblasts, keratinocytes and melanocytes. PMID:26308443

Soft 3D-Printed Phantom of the Human Kidney with Collecting System.

PubMed

Adams, Fabian; Qiu, Tian; Mark, Andrew; Fritz, Benjamin; Kramer, Lena; Schlager, Daniel; Wetterauer, Ulrich; Miernik, Arkadiusz; Fischer, Peer

2017-04-01

Organ models are used for planning and simulation of operations, developing new surgical instruments, and training purposes. There is a substantial demand for in vitro organ phantoms, especially in urological surgery. Animal models and existing simulator systems poorly mimic the detailed morphology and the physical properties of human organs. In this paper, we report a novel fabrication process to make a human kidney phantom with realistic anatomical structures and physical properties. The detailed anatomical structure was directly acquired from high resolution CT data sets of human cadaveric kidneys. The soft phantoms were constructed using a novel technique that combines 3D wax printing and polymer molding. Anatomical details and material properties of the phantoms were validated in detail by CT scan, ultrasound, and endoscopy. CT reconstruction, ultrasound examination, and endoscopy showed that the designed phantom mimics a real kidney's detailed anatomy and correctly corresponds to the targeted human cadaver's upper urinary tract. Soft materials with a tensile modulus of 0.8-1.5 MPa as well as biocompatible hydrogels were used to mimic human kidney tissues. We developed a method of constructing 3D organ models from medical imaging data using a 3D wax printing and molding process. This method is cost-effective means for obtaining a reproducible and robust model suitable for surgical simulation and training purposes.

Cytotoxic activity of the twigs of Cinnamomum cassia through the suppression of cell proliferation and the induction of apoptosis in human colorectal cancer cells.

PubMed

Park, Gwang Hun; Song, Hun Min; Park, Su Bin; Son, Ho-Jun; Um, Yurry; Kim, Hyun-Seok; Jeong, Jin Boo

2018-01-25

Because twigs of Cinnamomum cassia (TC) have been reported to exert anti-cancer activity, the mechanistic study for TC's anti-cancer activity is required. Thus, we elucidated the potential molecular mechanism of TC's anti-proliferative effect and the induction of apoptosis in human colorectal cancer cells. How water extracts form TC (TC-HW) was used in this study. Anti-cell proliferative effect of TC-HW was evaluated by MTT assay. The change of protein or mRNA level by TC-HW was evaluated by Western blot and RT-RCR, respectively. The promoter construct for ATF3, NF-κB, TOP-FLASH or FOP-FLASH was used for the investigation of the transcriptional activity for ATF3, NF-κB or Wnt. siRNA for ATF3 or p65 was used for the knockdown of ATF3 and p65. TC-HW reduced the cell viability in human colorectal cancer cells. TC-HW decreased cyclin D1 protein level through cyclin D1 degradation via GSK3β-dependent threonine-286 (T286) phosphorylation of cyclin D1, indicating that cyclin D1 degradation may contribute to TC-HW-mediated decrease of cyclin D1 protein level. TC-HW downregulated the expression of cyclin D1 mRNA level and inhibited Wnt activation through the downregulation of β-catenin and TCF4 expression, indicating that inhibition of cyclin D1 transcription may also result in TC-HW-mediated decrease of cyclin D1 protein level. In addition, TC-HW was observed to induce apoptosis through ROS-dependent DNA damage. TC-HW-induced ROS increased NF-κB and ATF3 activation, and inhibition of NF-κB and ATF3 activation attenuated TC-HW-mediated apoptosis. Our results suggest that TC-HW may suppress cell proliferation through the downregulation of cyclin D1 via proteasomal degradation and transcriptional inhibition, and may induce apoptosis through ROS-dependent NF-κB and ATF3 activation. These effects of TC-HW may contribute to the reduction of cell viability in human colorectal cancer cells. From these findings, TC-HW has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer.

The role of the innate immune system in destruction of pancreatic beta cells in NOD mice and humans with type I diabetes

PubMed Central

Tai, Ningwen; Wong, F. Susan; Wen, Li

2016-01-01

Type 1 diabetes (T1D) is an organ-specific autoimmune disease characterized by T cell-mediated destruction of the insulin-producing pancreatic β cells. A combination of genetic and environmental factors eventually leads to the loss of functional β cells mass and hyperglycemia. Both innate and adaptive immunity are involved in the development of T1D. In this review, we have highlighted the most recent findings on the role of innate immunity, especially the pattern recognition receptors (PRRs), in disease development. In murine models and human studies, different PRRs, such as toll-like receptors (TLRs) and nucleotide-binding domain, leucine-rich repeat-containing (or NOD-like) receptors (NLRs), have different roles in the pathogenesis of T1D. These PRRs play a critical role in defending against infection by sensing specific ligands derived from exogenous microorganisms to induce innate immune responses and shape adaptive immunity. Animal studies have shown that TLR7, TLR9, MyD88 and NLPR3 play a disease-predisposing role in T1D, while controversial results have been found with other PRRs, such as TLR2, TLR3, TLR4, TLR5 and others. Human studies also shown that TLR2, TLR3 and TLR4 are expressed in either islet β cells or infiltrated immune cells, indicating the innate immunity plays a role in β cell autoimmunity. Furthermore, some human genetic studies showed a possible association of TLR3, TLR7, TLR8 or NLRP3 genes, at single nucleotide polymorphism (SNP) level, with human T1D. Increasing evidence suggest that the innate immunity modulates β cell autoimmunity. Thus, targeting pathways of innate immunity may provide novel therapeutic strategies to fight this disease. PMID:27021275

3D Functional Corneal Stromal Tissue Equivalent Based on Corneal Stromal Stem Cells and Multi-Layered Silk Film Architecture.

PubMed

Ghezzi, Chiara E; Marelli, Benedetto; Omenetto, Fiorenzo G; Funderburgh, James L; Kaplan, David L

2017-01-01

The worldwide need for human cornea equivalents continues to grow. Few clinical options are limited to allogenic and synthetic material replacements. We hypothesized that tissue engineered human cornea systems based on mechanically robust, patterned, porous, thin, optically clear silk protein films, in combination with human corneal stromal stem cells (hCSSCs), would generate 3D functional corneal stroma tissue equivalents, in comparison to previously developed 2D approaches. Silk film contact guidance was used to control the alignment and distribution of hCSSCs on RGD-treated single porous silk films, which were then stacked in an orthogonally, multi-layered architecture and cultured for 9 weeks. These systems were compared similar systems generated with human corneal fibroblasts (hCFs). Both cell types were viable and preferentially aligned along the biomaterial patterns for up to 9 weeks in culture. H&E histological sections showed that the systems seeded with the hCSSCs displayed ECM production throughout the entire thickness of the constructs. In addition, the ECM proteins tested positive for keratocyte-specific tissue markers, including keratan sulfate, lumican, and keratocan. The quantification of hCSSC gene expression of keratocyte-tissue markers, including keratocan, lumican, human aldehyde dehydrogenase 3A1 (ALDH3A1), prostaglandin D2 synthase (PTDGS), and pyruvate dehydrogenase kinase, isozyme 4 (PDK4), within the 3D tissue systems demonstrated upregulation when compared to 2D single silk films and to the systems generated with the hCFs. Furthermore, the production of ECM from the hCSSC seeded systems and subsequent remodeling of the initial matrix significantly improved cohesiveness and mechanical performance of the constructs, while maintaining transparency after 9 weeks.

Characterization of [3H]LS-3-134, a Novel Arylamide Phenylpiperazine D3 Dopamine Receptor Selective Radioligand

PubMed Central

Rangel-Barajas, Claudia; Malik, Maninder; Taylor, Michelle; Neve, Kim A.; Mach, Robert H.; Luedtke, Robert R.

2014-01-01

LS-3-134 is a substituted N-phenylpiperazine derivative that has been reported to exhibit a) high-affinity binding (Ki value 0.2 nM) at human D3 dopamine receptors, b) >100-fold D3 vs. D2 dopamine receptor subtype binding selectivity and c) low-affinity binding (Ki values >5,000 nM) at sigma 1 and sigma 2 receptors. Based upon a forskolin-dependent activation of the adenylyl cyclase inhibition assay, LS-3-134 is a weak partial agonist at both D2 and D3 dopamine receptor subtypes (29% and 35% of full agonist activity, respectively). In this study, [3H]-labeled LS-3-134 was prepared and evaluated to further characterize its use as a D3 dopamine receptor selective radioligand. Kinetic and equilibrium radioligand binding studies were performed. This radioligand rapidly reaches equilibrium (10-15 min at 37°C) and binds with high affinity to both human (Kd = 0.06 ± 0.01 nM) and rat (Kd = 0.2 ± 0.02 nM) D3 receptors expressed in HEK-293 cells. Direct and competitive radioligand binding studies using rat caudate and nucleus accumbens tissue indicate that [3H]LS-3-134 selectively binds a homogeneous population of binding sites with a dopamine D3 receptor pharmacological profile. Based upon these studies we propose that [3H]LS-3-134 represents a novel D3 dopamine receptor selective radioligand that can be used for studying the expression and regulation of the D3 dopamine receptor subtype. PMID:25041389

Engineered cell and tissue models of pulmonary fibrosis.

PubMed

Sundarakrishnan, Aswin; Chen, Ying; Black, Lauren D; Aldridge, Bree B; Kaplan, David L

2018-04-01

Pulmonary fibrosis includes several lung disorders characterized by scar formation and Idiopathic Pulmonary Fibrosis (IPF) is a particularly severe form of pulmonary fibrosis of unknown etiology with a mean life expectancy of 3years' post-diagnosis. Treatments for IPF are limited to two FDA approved drugs, pirfenidone and nintedanib. Most lead candidate drugs that are identified in pre-clinical animal studies fail in human clinical trials. Thus, there is a need for advanced humanized in vitro models of the lung to improve candidate treatments prior to moving to human clinical trials. The development of 3D tissue models has created systems capable of emulating human lung structure, function, and cell and matrix interactions. The specific models accomplish these features and preliminary studies conducted using some of these systems have shown potential for in vitro anti-fibrotic drug testing. Further characterization and improvements will enable these tissue models to extend their utility for in vitro drug testing, to help identify signaling pathways and mechanisms for new drug targets, and potentially reduce animal models as standard pre-clinical models of study. In the current review, we contrast different in vitro models based on increasing dimensionality (2D, 2.5D and 3D), with added focus on contemporary 3D pulmonary models of fibrosis. Copyright © 2017. Published by Elsevier B.V.

Benchmark datasets for 3D MALDI- and DESI-imaging mass spectrometry.

PubMed

Oetjen, Janina; Veselkov, Kirill; Watrous, Jeramie; McKenzie, James S; Becker, Michael; Hauberg-Lotte, Lena; Kobarg, Jan Hendrik; Strittmatter, Nicole; Mróz, Anna K; Hoffmann, Franziska; Trede, Dennis; Palmer, Andrew; Schiffler, Stefan; Steinhorst, Klaus; Aichler, Michaela; Goldin, Robert; Guntinas-Lichius, Orlando; von Eggeling, Ferdinand; Thiele, Herbert; Maedler, Kathrin; Walch, Axel; Maass, Peter; Dorrestein, Pieter C; Takats, Zoltan; Alexandrov, Theodore

2015-01-01

Three-dimensional (3D) imaging mass spectrometry (MS) is an analytical chemistry technique for the 3D molecular analysis of a tissue specimen, entire organ, or microbial colonies on an agar plate. 3D-imaging MS has unique advantages over existing 3D imaging techniques, offers novel perspectives for understanding the spatial organization of biological processes, and has growing potential to be introduced into routine use in both biology and medicine. Owing to the sheer quantity of data generated, the visualization, analysis, and interpretation of 3D imaging MS data remain a significant challenge. Bioinformatics research in this field is hampered by the lack of publicly available benchmark datasets needed to evaluate and compare algorithms. High-quality 3D imaging MS datasets from different biological systems at several labs were acquired, supplied with overview images and scripts demonstrating how to read them, and deposited into MetaboLights, an open repository for metabolomics data. 3D imaging MS data were collected from five samples using two types of 3D imaging MS. 3D matrix-assisted laser desorption/ionization imaging (MALDI) MS data were collected from murine pancreas, murine kidney, human oral squamous cell carcinoma, and interacting microbial colonies cultured in Petri dishes. 3D desorption electrospray ionization (DESI) imaging MS data were collected from a human colorectal adenocarcinoma. With the aim to stimulate computational research in the field of computational 3D imaging MS, selected high-quality 3D imaging MS datasets are provided that could be used by algorithm developers as benchmark datasets.

3D-printing and the effect on medical costs: a new era?

PubMed

Choonara, Yahya E; du Toit, Lisa C; Kumar, Pradeep; Kondiah, Pierre P D; Pillay, Viness

2016-01-01

3D-printing (3DP) is the art and science of printing in a new dimension using 3D printers to transform 3D computer aided designs (CAD) into life-changing products. This includes the design of more effective and patient-friendly pharmaceutical products as well as bio-inspired medical devices. It is poised as the next technology revolution for the pharmaceutical and medical-device industries. After decorous implementation scientists in collaboration with CAD designers have produced innovative medical devices ranging from pharmaceutical tablets to surgical transplants of the human face and skull, spinal implants, prosthetics, human organs and other biomaterials. While 3DP may be cost-efficient, a limitation exists in the availability of 3D printable biomaterials for most applications. In addition, the loss of skilled labor in producing medical devices such as prosthetics and other devices may affect developing economies. This review objectively explores the potential growth and impact of 3DP costs in the medical industry.

Effects of bright light exposure during daytime on peripheral clock gene expression in humans.

PubMed

Sato, Maki; Wakamura, Tomoko; Morita, Takeshi; Okamoto, Akihiko; Akashi, Makoto; Matsui, Takuya; Sato, Motohiko

2017-06-01

Light is the strongest synchronizer controlling circadian rhythms. The intensity and duration of light change throughout the year, thereby influencing body weight, food preferences, and melatonin secretion in humans and animals. Although the expression of clock genes has been examined using human samples, it currently remains unknown whether bright light during the daytime affects the expression of these genes in humans. Therefore, we herein investigated the effects of bright light exposure during the daytime on clock gene expression in the hair follicular and root cells of the human scalp. Seven healthy men (20.4 ± 2.2 years old; 172.3 ± 5.8 cm; 64.3 ± 8.5 kg; BMI 21.7 ± 3.1 kg/m 2 , mean ± SD) participated in this study. Subjects completed 3-day experimental sessions twice in 1 month during which they were exposed to bright and dim light conditions. The mRNA expression of Per1-3, Cry1-2, Rev-erb-α (Nr1d1), Rev-erb-β (Nr1d2), and Dec1 was analyzed using branched DNA probes. No significant changes were observed in the expression of Per1, Per2, Per3, Cry1, Cry2, Rev-erb-α (Nr1d1), or Dec1 following exposure to bright light conditions. However, the expression of Rev-erb-β (Nr1d2) tended to be stronger under bright light than dim light conditions. These results suggest that the bright light stimulus did not influence the expression of clock genes in humans. Long-lasting bright light exposure during the daytime may be required to change the expression of clock genes in humans.

Phenylbutyrate Is Bacteriostatic against Mycobacterium tuberculosis and Regulates the Macrophage Response to Infection, Synergistically with 25-Hydroxy-Vitamin D₃

PubMed Central

Coussens, Anna K.; Wilkinson, Robert J.; Martineau, Adrian R.

2015-01-01

Adjunctive vitamin D treatment for pulmonary tuberculosis enhances resolution of inflammation but has modest effects on bacterial clearance. Sodium 4-phenylbutyrate (PBA) is in clinical use for a range of conditions and has been shown to synergise with vitamin D metabolites to upregulate cathelicidin antimicrobial peptide (CAMP) expression. We investigated whether clinically attainable plasma concentrations of PBA (0.4-4mM) directly affect Mycobacterium tuberculosis (Mtb) growth and human macrophage and PBMC response to infection. We also tested the ability of PBA to enhance the immunomodulatory actions of the vitamin D metabolite 25(OH)D3 during infection and synergistically inhibit intracellular Mtb growth. PBA inhibited Mtb growth in broth with an MIC99 of 1mM, which was reduced to 0.25mM by lowering pH. During human macrophage infection, PBA treatment restricted Mtb uptake, phagocytic receptor expression and intracellular growth in a dose-dependent manner. PBA independently regulated CCL chemokine secretion and induced expression of the antimicrobial LTF (lactoferrin), the anti-inflammatory PROC (protein C) and multiple genes within the NLRP3 inflammasome pathway. PBA co-treatment with 25(OH)D3 synergistically modulated expression of numerous vitamin D-response genes, including CAMP, CYP24A1, CXCL10 and IL-37. This synergistic effect was dependent on MAPK signalling, while the effect of PBA on LTF, PROC and NLRP3 was MAPK-independent. During PBA and 25(OH)D3 co-treatment of human macrophages, in the absence of exogenous proteinase 3 (PR3) to activate cathelicidin, Mtb growth restriction was dominated by the effect of PBA, while the addition of PR3 enhanced growth restriction by 25(OH)D3 and PBA co-treatment. This suggests that PBA augments vitamin D–mediated cathelicidin-dependent Mtb growth restriction by human macrophages and independently induces antimicrobial and anti-inflammatory action. Therefore through both host-directed and bacterial-directed mechanisms PBA and vitamin D may prove an effective combinatorial adjunct therapy for tuberculosis to both resolve immunopathology and enhance bacterial clearance. PMID:26133770

My Corporis Fabrica Embryo: An ontology-based 3D spatio-temporal modeling of human embryo development.

PubMed

Rabattu, Pierre-Yves; Massé, Benoit; Ulliana, Federico; Rousset, Marie-Christine; Rohmer, Damien; Léon, Jean-Claude; Palombi, Olivier

2015-01-01

Embryology is a complex morphologic discipline involving a set of entangled mechanisms, sometime difficult to understand and to visualize. Recent computer based techniques ranging from geometrical to physically based modeling are used to assist the visualization and the simulation of virtual humans for numerous domains such as surgical simulation and learning. On the other side, the ontology-based approach applied to knowledge representation is more and more successfully adopted in the life-science domains to formalize biological entities and phenomena, thanks to a declarative approach for expressing and reasoning over symbolic information. 3D models and ontologies are two complementary ways to describe biological entities that remain largely separated. Indeed, while many ontologies providing a unified formalization of anatomy and embryology exist, they remain only descriptive and make the access to anatomical content of complex 3D embryology models and simulations difficult. In this work, we present a novel ontology describing the development of the human embryology deforming 3D models. Beyond describing how organs and structures are composed, our ontology integrates a procedural description of their 3D representations, temporal deformation and relations with respect to their developments. We also created inferences rules to express complex connections between entities. It results in a unified description of both the knowledge of the organs deformation and their 3D representations enabling to visualize dynamically the embryo deformation during the Carnegie stages. Through a simplified ontology, containing representative entities which are linked to spatial position and temporal process information, we illustrate the added-value of such a declarative approach for interactive simulation and visualization of 3D embryos. Combining ontologies and 3D models enables a declarative description of different embryological models that capture the complexity of human developmental anatomy. Visualizing embryos with 3D geometric models and their animated deformations perhaps paves the way towards some kind of hypothesis-driven application. These can also be used to assist the learning process of this complex knowledge. http://www.mycorporisfabrica.org/.

Alternatives to In Vivo Draize Rabbit Eye and Skin Irritation Tests with a Focus on 3D Reconstructed Human Cornea-Like Epithelium and Epidermis Models

PubMed Central

Lee, Miri; Hwang, Jee-Hyun; Lim, Kyung-Min

2017-01-01

Human eyes and skin are frequently exposed to chemicals accidentally or on purpose due to their external location. Therefore, chemicals are required to undergo the evaluation of the ocular and dermal irritancy for their safe handling and use before release into the market. Draize rabbit eye and skin irritation test developed in 1944, has been a gold standard test which was enlisted as OECD TG 404 and OECD TG 405 but it has been criticized with respect to animal welfare due to invasive and cruel procedure. To replace it, diverse alternatives have been developed: (i) For Draize eye irritation test, organotypic assay, in vitro cytotoxicity-based method, in chemico tests, in silico prediction model, and 3D reconstructed human cornea-like epithelium (RhCE); (ii) For Draize skin irritation test, in vitro cytotoxicity-based cell model, and 3D reconstructed human epidermis models (RhE). Of these, RhCE and RhE models are getting spotlight as a promising alternative with a wide applicability domain covering cosmetics and personal care products. In this review, we overviewed the current alternatives to Draize test with a focus on 3D human epithelium models to provide an insight into advancing and widening their utility. PMID:28744350

5D-intravital tomography as a novel tool for non-invasive in-vivo analysis of human skin

NASA Astrophysics Data System (ADS)

König, Karsten; Weinigel, Martin; Breunig, Hans G.; Gregory, Axel; Fischer, Peter; Kellner-Höfer, Marcel; Bückle, Rainer; Schwarz, Martin; Riemann, Iris; Stracke, Frank; Huck, Volker; Gorzelanny, Christian; Schneider, Stefan W.

2010-02-01

Some years ago, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have been launched. These tomographs provide optical biopsies with submicron resolution based on two-photon excited autofluorescence (NAD(P)H, flavoproteins, keratin, elastin, melanin, porphyrins) and second harmonic generation by collagen. The 3D tomograph was now transferred into a 5D imaging system by the additional detection of the emission spectrum and the fluorescence lifetime based on spatially and spectrally resolved time-resolved single photon counting. The novel 5D intravital tomograph (5D-IVT) was employed for the early detection of atopic dermatitis and the analysis of treatment effects.

Combining heterogenous features for 3D hand-held object recognition

NASA Astrophysics Data System (ADS)

Lv, Xiong; Wang, Shuang; Li, Xiangyang; Jiang, Shuqiang

2014-10-01

Object recognition has wide applications in the area of human-machine interaction and multimedia retrieval. However, due to the problem of visual polysemous and concept polymorphism, it is still a great challenge to obtain reliable recognition result for the 2D images. Recently, with the emergence and easy availability of RGB-D equipment such as Kinect, this challenge could be relieved because the depth channel could bring more information. A very special and important case of object recognition is hand-held object recognition, as hand is a straight and natural way for both human-human interaction and human-machine interaction. In this paper, we study the problem of 3D object recognition by combining heterogenous features with different modalities and extraction techniques. For hand-craft feature, although it reserves the low-level information such as shape and color, it has shown weakness in representing hiconvolutionalgh-level semantic information compared with the automatic learned feature, especially deep feature. Deep feature has shown its great advantages in large scale dataset recognition but is not always robust to rotation or scale variance compared with hand-craft feature. In this paper, we propose a method to combine hand-craft point cloud features and deep learned features in RGB and depth channle. First, hand-held object segmentation is implemented by using depth cues and human skeleton information. Second, we combine the extracted hetegerogenous 3D features in different stages using linear concatenation and multiple kernel learning (MKL). Then a training model is used to recognize 3D handheld objects. Experimental results validate the effectiveness and gerneralization ability of the proposed method.

Bioprinting of 3D Convoluted Renal Proximal Tubules on Perfusable Chips

NASA Astrophysics Data System (ADS)

Homan, Kimberly A.; Kolesky, David B.; Skylar-Scott, Mark A.; Herrmann, Jessica; Obuobi, Humphrey; Moisan, Annie; Lewis, Jennifer A.

2016-10-01

Three-dimensional models of kidney tissue that recapitulate human responses are needed for drug screening, disease modeling, and, ultimately, kidney organ engineering. Here, we report a bioprinting method for creating 3D human renal proximal tubules in vitro that are fully embedded within an extracellular matrix and housed in perfusable tissue chips, allowing them to be maintained for greater than two months. Their convoluted tubular architecture is circumscribed by proximal tubule epithelial cells and actively perfused through the open lumen. These engineered 3D proximal tubules on chip exhibit significantly enhanced epithelial morphology and functional properties relative to the same cells grown on 2D controls with or without perfusion. Upon introducing the nephrotoxin, Cyclosporine A, the epithelial barrier is disrupted in a dose-dependent manner. Our bioprinting method provides a new route for programmably fabricating advanced human kidney tissue models on demand.

Bioprinting of 3D Convoluted Renal Proximal Tubules on Perfusable Chips

PubMed Central

Homan, Kimberly A.; Kolesky, David B.; Skylar-Scott, Mark A.; Herrmann, Jessica; Obuobi, Humphrey; Moisan, Annie; Lewis, Jennifer A.

2016-01-01

Three-dimensional models of kidney tissue that recapitulate human responses are needed for drug screening, disease modeling, and, ultimately, kidney organ engineering. Here, we report a bioprinting method for creating 3D human renal proximal tubules in vitro that are fully embedded within an extracellular matrix and housed in perfusable tissue chips, allowing them to be maintained for greater than two months. Their convoluted tubular architecture is circumscribed by proximal tubule epithelial cells and actively perfused through the open lumen. These engineered 3D proximal tubules on chip exhibit significantly enhanced epithelial morphology and functional properties relative to the same cells grown on 2D controls with or without perfusion. Upon introducing the nephrotoxin, Cyclosporine A, the epithelial barrier is disrupted in a dose-dependent manner. Our bioprinting method provides a new route for programmably fabricating advanced human kidney tissue models on demand. PMID:27725720

The Influence on Humans of Long Hours of Viewing 3D Movies

NASA Astrophysics Data System (ADS)

Kawamura, Yuta; Horie, Yusuke; Sano, Keisuke; Kodama, Hiroya; Tsunoda, Naoki; Shibuta, Yuki; Kawachi, Yuki; Yamada, Mitsuho

Three-dimensional (3D) movies have become very popular in movie theaters and for home viewing, To date, there has been no report of the effects of the continual vergence eye movement that occurs when viewing 3D movies from the beginning to the end. First, we analyzed the influence of viewing a 3D movie for several hours on vergence eye movement. At the same time, we investigated the influence of long viewing on the human body, using the Simulator Sickness Questionnaire (SSQ) and critical fusion frequency (CFF). It was suggested that the vergence stable time after saccade when viewing a long movie was influenced by the viewing time and that the vergence stable time after saccade depended on the content of the movie. Also the differences were seen in the SSQ and CFF between the movie's beginning and its ending when viewing a 3D movie.

3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes.

PubMed

Correia, Cláudia; Koshkin, Alexey; Duarte, Patrícia; Hu, Dongjian; Carido, Madalena; Sebastião, Maria J; Gomes-Alves, Patrícia; Elliott, David A; Domian, Ibrahim J; Teixeira, Ana P; Alves, Paula M; Serra, Margarida

2018-03-01

Three-dimensional (3D) cultures of human pluripotent stem cell derived cardiomyocytes (hPSC-CMs) hold great promise for drug discovery, providing a better approximation to the in vivo physiology over standard two-dimensional (2D) monolayer cultures. However, the transition of CM differentiation protocols from 2D to 3D cultures is not straightforward. In this work, we relied on the aggregation of hPSC-derived cardiac progenitors and their culture under agitated conditions to generate highly pure cardiomyocyte aggregates. Whole-transcriptome analysis and 13 C-metabolic flux analysis allowed to demonstrate at both molecular and fluxome levels that such 3D culture environment enhances metabolic maturation of hiPSC-CMs. When compared to 2D, 3D cultures of hiPSC-CMs displayed down-regulation of genes involved in glycolysis and lipid biosynthesis and increased expression of genes involved in OXPHOS. Accordingly, 3D cultures of hiPSC-CMs had lower fluxes through glycolysis and fatty acid synthesis and increased TCA-cycle activity. Importantly, we demonstrated that the 3D culture environment reproducibly improved both CM purity and metabolic maturation across different hPSC lines, thereby providing a robust strategy to derive enriched hPSC-CMs with metabolic features closer to that of adult CMs. © 2017 Wiley Periodicals, Inc.

Highly potent analogues of luteinizing hormone-releasing hormone containing D-phenylalanine nitrogen mustard in position 6

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bajusz, S.; Janaky, T.; Csernus, V.J.

The nitrogen mustard derivatives of 4-phenylbutyric acid and L-phenylalanine, called chlorambucil (Chl) and melphalan (Mel), respectively, have been incorporated into several peptide hormones, including luteinizing hormone-releasing hormone (LH-RH). The alkylating analogues of LH-RH were prepared by linking Chl, as an N-acyl moiety, to the complete amino acid sequence of agonistic and antagonistic analogues. These compounds, in particular the antagonistic analogues, showed much lower potency than their congeners carrying other acyl groups. To obtain highly potent alkylating analogues of LH-RH, the D enantiomer of Mel was incorporated into position 6 of the native hormone and some of its antagonistic analogues. Ofmore » the peptides prepared, (D-Mel{sup 6})LH-RH (SB-05) and (Ac-D-Nal(2){sup 1},D-Phe(pCl){sup 2},D-Pal(3){sup 3},Arg{sup 5},D-Mel{sup 6},D-Ala{sup 10})LH-RH (SB-86, where Nal(2) is 3-(2-naphthyl)alanine and Pal(3) is 3-(3-pyridyl)alanine) possessed the expected high agonistic and antagonistic activities, respectively, and also showed high affinities for the membrane receptors of rat pituitary cells, human breast cancer cells, human prostate cancer cells, and rat Dunning R-3327 prostate tumor cells. These two analogues exerted cytotoxic effects on human and rat mammary cancer cells in vitro. Thus these two D-Mel{sup 6} analogues seem to be particularly suitable for the study of how alkylating analogues of LH-RH could interfere with intracellular events in certain cancer cells.« less

Reduced bone resorption by intake of dietary vitamin D and K from tailor-made Atlantic salmon: a randomized intervention trial

PubMed Central

Graff, Ingvild Eide; Øyen, Jannike; Kjellevold, Marian; Frøyland, Livar; Gjesdal, Clara Gram; Almås, Bjørg; Rosenlund, Grethe; Lie, Øyvind

2016-01-01

Suboptimal vitamin D status is common among humans, and might increase bone resorption with subsequent negative effects on bone health. Fatty fish, including Atlantic salmon, is an important dietary vitamin D source. However, due to a considerable change in fish feed composition, the contribution of vitamin D from salmon fillet has been reduced. The main objective was to investigate if intake of vitamin D3 enriched salmon or vitamin D3 tablets decreased bone biomarkers (urinary N-telopeptides, deoxypyridinoline, serum bone-specific alkaline phosphatase, and osteocalcin) compared to a low vitamin D3 intake. The 122 healthy postmenopausal women included in this 12 weeks intervention trial were randomized into four groups: three salmon groups (150 grams/two times/week) and one tablet group (800 IU vitamin D and 1000 mg calcium/day). The salmon groups also received calcium supplements. The salmon had three different vitamin D3/vitamin K1 combinations: high D3+high K1, low D3+high K1, or high D3+low K1. Increased intake of salmon containing high levels of vitamin D3 (0.35-0.38 mg/kg/fillet) and supplements with the same weekly contribution had a positive influence on bone health as measured by bone biomarkers in postmenopausal women. Consequently, an increased level of vitamin D3 at least to original level in feed for salmonids will contribute to an improved vitamin D3 status and may improve human bone health. PMID:27542236

3D morphology-based clustering and simulation of human pyramidal cell dendritic spines.

PubMed

Luengo-Sanchez, Sergio; Fernaud-Espinosa, Isabel; Bielza, Concha; Benavides-Piccione, Ruth; Larrañaga, Pedro; DeFelipe, Javier

2018-06-13

The dendritic spines of pyramidal neurons are the targets of most excitatory synapses in the cerebral cortex. They have a wide variety of morphologies, and their morphology appears to be critical from the functional point of view. To further characterize dendritic spine geometry, we used in this paper over 7,000 individually 3D reconstructed dendritic spines from human cortical pyramidal neurons to group dendritic spines using model-based clustering. This approach uncovered six separate groups of human dendritic spines. To better understand the differences between these groups, the discriminative characteristics of each group were identified as a set of rules. Model-based clustering was also useful for simulating accurate 3D virtual representations of spines that matched the morphological definitions of each cluster. This mathematical approach could provide a useful tool for theoretical predictions on the functional features of human pyramidal neurons based on the morphology of dendritic spines.

Super Normal Vector for Human Activity Recognition with Depth Cameras.

PubMed

Yang, Xiaodong; Tian, YingLi

2017-05-01

The advent of cost-effectiveness and easy-operation depth cameras has facilitated a variety of visual recognition tasks including human activity recognition. This paper presents a novel framework for recognizing human activities from video sequences captured by depth cameras. We extend the surface normal to polynormal by assembling local neighboring hypersurface normals from a depth sequence to jointly characterize local motion and shape information. We then propose a general scheme of super normal vector (SNV) to aggregate the low-level polynormals into a discriminative representation, which can be viewed as a simplified version of the Fisher kernel representation. In order to globally capture the spatial layout and temporal order, an adaptive spatio-temporal pyramid is introduced to subdivide a depth video into a set of space-time cells. In the extensive experiments, the proposed approach achieves superior performance to the state-of-the-art methods on the four public benchmark datasets, i.e., MSRAction3D, MSRDailyActivity3D, MSRGesture3D, and MSRActionPairs3D.

Fusion of Building Information and Range Imaging for Autonomous Location Estimation in Indoor Environments

PubMed Central

Kohoutek, Tobias K.; Mautz, Rainer; Wegner, Jan D.

2013-01-01

We present a novel approach for autonomous location estimation and navigation in indoor environments using range images and prior scene knowledge from a GIS database (CityGML). What makes this task challenging is the arbitrary relative spatial relation between GIS and Time-of-Flight (ToF) range camera further complicated by a markerless configuration. We propose to estimate the camera's pose solely based on matching of GIS objects and their detected location in image sequences. We develop a coarse-to-fine matching strategy that is able to match point clouds without any initial parameters. Experiments with a state-of-the-art ToF point cloud show that our proposed method delivers an absolute camera position with decimeter accuracy, which is sufficient for many real-world applications (e.g., collision avoidance). PMID:23435055

Multidirectional four-dimensional shape measurement system

NASA Astrophysics Data System (ADS)

Lenar, Janusz; Sitnik, Robert; Witkowski, Marcin

2012-03-01

Currently, a lot of different scanning techniques are used for 3D imaging of human body. Most of existing systems are based on static registration of internal structures using MRI or CT techniques as well as 3D scanning of outer surface of human body by laser triangulation or structured light methods. On the other hand there is an existing mature 4D method based on tracking in time the position of retro-reflective markers attached to human body. There are two main drawbacks of this solution: markers are attached to skin (no real skeleton movement is registered) and it gives (x, y, z, t) coordinates only in those points (not for the whole surface). In this paper we present a novel multidirectional structured light measurement system that is capable of measuring 3D shape of human body surface with frequency reaching 60Hz. The developed system consists of two spectrally separated and hardware-synchronized 4D measurement heads. The principle of the measurement is based on single frame analysis. Projected frame is composed from sine-modulated intensity pattern and a special stripe allowing absolute phase measurement. Several different geometrical set-ups will be proposed depending on type of movements that are to be registered.

Evolutionary history of linked D4Z4 and Beta satellite clusters at the FSHD locus (4q35)☆

PubMed Central

Giussani, Marta; Cardone, Maria Francesca; Bodega, Beatrice; Ginelli, Enrico; Meneveri, Raffaella

2012-01-01

We performed a detailed genomic investigation of the chimpanzee locus syntenic to human chromosome 4q35.2, associated to the facioscapulohumeral dystrophy. Two contigs of approximately 150 kb and 200 kb were derived from PTR chromosomes 4q35 and 3p12, respectively: both regions showed a very similar sequence organization, including D4Z4 and Beta satellite linked clusters. Starting from these findings, we derived a hypothetical evolutionary history of human 4q35, 10q26 and 3p12 chromosome regions focusing on the D4Z4–Beta satellite linked organization. The D4Z4 unit showed an open reading frame (DUX4) at both PTR 4q35 and 3p12 regions; furthermore some subregions of the Beta satellite unit showed a high degree of conservation between chimpanzee and humans. In conclusion, this paper provides evidence that at the 4q subtelomere the linkage between D4Z4 and Beta satellite arrays is a feature that appeared late during evolution and is conserved between chimpanzee and humans. PMID:22824653

Evolutionary history of linked D4Z4 and Beta satellite clusters at the FSHD locus (4q35).

PubMed

Giussani, Marta; Cardone, Maria Francesca; Bodega, Beatrice; Ginelli, Enrico; Meneveri, Raffaella

2012-11-01

We performed a detailed genomic investigation of the chimpanzee locus syntenic to human chromosome 4q35.2, associated to the facioscapulohumeral dystrophy. Two contigs of approximately 150 kb and 200 kb were derived from PTR chromosomes 4q35 and 3p12, respectively: both regions showed a very similar sequence organization, including D4Z4 and Beta satellite linked clusters. Starting from these findings, we derived a hypothetical evolutionary history of human 4q35, 10q26 and 3p12 chromosome regions focusing on the D4Z4-Beta satellite linked organization. The D4Z4 unit showed an open reading frame (DUX4) at both PTR 4q35 and 3p12 regions; furthermore some subregions of the Beta satellite unit showed a high degree of conservation between chimpanzee and humans. In conclusion, this paper provides evidence that at the 4q subtelomere the linkage between D4Z4 and Beta satellite arrays is a feature that appeared late during evolution and is conserved between chimpanzee and humans. Copyright © 2012 Elsevier Inc. All rights reserved.

Cytochrome P450 isoforms in the Metabolism of Decursin and Decursinol Angelate from Korean Angelica

PubMed Central

ZHANG, Jinhui; LI, Li; TANG, Suni; HALE, Thomas W.; XING, Chengguo; JIANG, Cheng; LÜ, Junxuan

2016-01-01

We have shown that the in vitro hepatic microsomal metabolism of pyranocoumarin compound decursinol angelate (DA) to decursinol (DOH) exclusively requires cytochrome P450 enzymes (CYP) whereas the conversion of its isomer decursin (D) to DOH can be mediated by CYP and esterase(s). To provide insight into specific isoforms involved, here we show with recombinant human CYP that 2C19 was the most active at metabolizing D and DA in vitro followed by 3A4. With carboxylesterases (CES), D was hydrolyzed by CES2 but not CES1, and DA was resistant to both CES1 and CES2. In human liver microsomal preparation, general CYP inhibitor 1-aminobenzotriazole (ABT) and respective competitive inhibitors for 2C19 and 3A4, (+)-N-3-benzylnirvanol and ketoconazole, substantially retarded the metabolism of DA and, to a lesser extent, of D. In healthy human subjects from a single-dose pharmacokinetic study, 2C19 extensive metabolizer genotype (2C19*17 allele) tended to have less plasma DA AUC0–48h and poor metabolizer genotype (2C19*2 allele) tended to have greater DA AUC0–48h. In mice given a single dose of D/DA, pretreatment with ABT boosted the plasma and prostate levels of D and DA by more than an order of magnitude. Taken together, our findings suggest that CYP isoforms 2C19 and 3A4 may play a crucial role in the first pass liver metabolism of DA and, to a lesser extent, that of D in humans. Pharmacogenetics with respect to CYP genotypes and interactions among CYP inhibitor drugs and D/DA should therefore be considered in designing future translation studies of DA and/or D. PMID:26394652

Cytochrome P450 Isoforms in the Metabolism of Decursin and Decursinol Angelate from Korean Angelica.

PubMed

Zhang, Jinhui; Li, Li; Tang, Suni; Hale, Thomas W; Xing, Chengguo; Jiang, Cheng; Lü, Junxuan

2015-01-01

We have shown that the in vitro hepatic microsomal metabolism of pyranocoumarin compound decursinol angelate (DA) to decursinol (DOH) exclusively requires cytochrome P450 (CYP) enzymes, whereas the conversion of its isomer decursin (D) to DOH can be mediated by CYP and esterase(s). To provide insight into specific isoforms involved, here we show with recombinant human CYP that 2C19 was the most active at metabolizing D and DA in vitro followed by 3A4. With carboxylesterases (CES), D was hydrolyzed by CES2 but not CES1, and DA was resistant to both CES1 and CES2. In human liver microsomal (HLM) preparation, the general CYP inhibitor 1-aminobenzotriazole (ABT) and respective competitive inhibitors for 2C19 and 3A4, (+)-N-3-benzylnirvanol (NBN) and ketoconazole substantially retarded the metabolism of DA and, to a lesser extent, of D. In healthy human subjects from a single-dose pharmacokinetic (PK) study, 2C19 extensive metabolizer genotype (2C19*17 allele) tended to have less plasma DA AUC0-48h and poor metabolizer genotype (2C19*2 allele) tended to have greater DA AUC0-48h. In mice given a single dose of D/DA, pretreatment with ABT boosted the plasma and prostate levels of D and DA by more than an order of magnitude. Taken together, our findings suggest that CYP isoforms 2C19 and 3A4 may play a crucial role in the first pass liver metabolism of DA and, to a lesser extent, that of D in humans. Pharmacogenetics with respect to CYP genotypes and interactions among CYP inhibitor drugs and D/DA should therefore be considered in designing future translation studies of DA and/or D.

Quadruplexes of human telomere dG{sub 3}(TTAG{sub 3}){sub 3} sequences containing guanine abasic sites

DOE Office of Scientific and Technical Information (OSTI.GOV)

Skolakova, Petra; Bednarova, Klara; Vorlickova, Michaela

Research highlights: {yields} Loss of a guanine base does not hinder the formation of G-quadruplex of human telomere sequence. {yields} Each depurination strongly destabilizes the quadruplex of dG{sub 3}(TTAG{sub 3}){sub 3} in NaCl and KCl. {yields} Conformational change of the abasic analogs of dG{sub 3}(TTAG{sub 3}){sub 3} is inhibited in KCl. {yields} The effects abasic sites may affect telomere-end structures in vivo. -- Abstract: This study was performed to evaluate how the loss of a guanine base affects the structure and stability of the three-tetrad G-quadruplex of 5'-dG{sub 3}(TTAG{sub 3}){sub 3}, the basic quadruplex-forming unit of the human telomere DNA.more » None of the 12 possible abasic sites hindered the formation of quadruplexes, but all reduced the thermodynamic stability of the parent quadruplex in both NaCl and KCl. The base loss did not change the Na{sup +}-stabilized intramolecular antiparallel architecture, based on CD spectra, but held up the conformational change induced in dG{sub 3}(TTAG{sub 3}){sub 3} in physiological concentration of KCl. The reduced stability and the inhibited conformational transitions observed here in vitro for the first time may predict that unrepaired abasic sites in G-quadruplexes could lead to changes in the chromosome's terminal protection in vivo.« less

Biomaterials-based 3D cell printing for next-generation therapeutics and diagnostics.

PubMed

Jang, Jinah; Park, Ju Young; Gao, Ge; Cho, Dong-Woo

2018-02-01

Building human tissues via 3D cell printing technology has received particular attention due to its process flexibility and versatility. This technology enables the recapitulation of unique features of human tissues and the all-in-one manufacturing process through the design of smart and advanced biomaterials and proper polymerization techniques. For the optimal engineering of tissues, a higher-order assembly of physiological components, including cells, biomaterials, and biomolecules, should meet the critical requirements for tissue morphogenesis and vascularization. The convergence of 3D cell printing with a microfluidic approach has led to a significant leap in the vascularization of engineering tissues. In addition, recent cutting-edge technology in stem cells and genetic engineering can potentially be adapted to the 3D tissue fabrication technique, and it has great potential to shift the paradigm of disease modeling and the study of unknown disease mechanisms required for precision medicine. This review gives an overview of recent developments in 3D cell printing and bioinks and provides technical requirements for engineering human tissues. Finally, we propose suggestions on the development of next-generation therapeutics and diagnostics. Copyright © 2017 Elsevier Ltd. All rights reserved.

Noninvasive, three-dimensional full-field body sensor for surface deformation monitoring of human body in vivo.

PubMed

Chen, Zhenning; Shao, Xinxing; He, Xiaoyuan; Wu, Jialin; Xu, Xiangyang; Zhang, Jinlin

2017-09-01

Noninvasive, three-dimensional (3-D), full-field surface deformation measurements of the human body are important for biomedical investigations. We proposed a 3-D noninvasive, full-field body sensor based on stereo digital image correlation (stereo-DIC) for surface deformation monitoring of the human body in vivo. First, by applying an improved water-transfer printing (WTP) technique to transfer optimized speckle patterns onto the skin, the body sensor was conveniently and harmlessly fabricated directly onto the human body. Then, stereo-DIC was used to achieve 3-D noncontact and noninvasive surface deformation measurements. The accuracy and efficiency of the proposed body sensor were verified and discussed by considering different complexions. Moreover, the fabrication of speckle patterns on human skin, which has always been considered a challenging problem, was shown to be feasible, effective, and harmless as a result of the improved WTP technique. An application of the proposed stereo-DIC-based body sensor was demonstrated by measuring the pulse wave velocity of human carotid artery. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Three-Dimensional Human iPSC-Derived Artificial Skeletal Muscles Model Muscular Dystrophies and Enable Multilineage Tissue Engineering.

PubMed

Maffioletti, Sara Martina; Sarcar, Shilpita; Henderson, Alexander B H; Mannhardt, Ingra; Pinton, Luca; Moyle, Louise Anne; Steele-Stallard, Heather; Cappellari, Ornella; Wells, Kim E; Ferrari, Giulia; Mitchell, Jamie S; Tyzack, Giulia E; Kotiadis, Vassilios N; Khedr, Moustafa; Ragazzi, Martina; Wang, Weixin; Duchen, Michael R; Patani, Rickie; Zammit, Peter S; Wells, Dominic J; Eschenhagen, Thomas; Tedesco, Francesco Saverio

2018-04-17

Generating human skeletal muscle models is instrumental for investigating muscle pathology and therapy. Here, we report the generation of three-dimensional (3D) artificial skeletal muscle tissue from human pluripotent stem cells, including induced pluripotent stem cells (iPSCs) from patients with Duchenne, limb-girdle, and congenital muscular dystrophies. 3D skeletal myogenic differentiation of pluripotent cells was induced within hydrogels under tension to provide myofiber alignment. Artificial muscles recapitulated characteristics of human skeletal muscle tissue and could be implanted into immunodeficient mice. Pathological cellular hallmarks of incurable forms of severe muscular dystrophy could be modeled with high fidelity using this 3D platform. Finally, we show generation of fully human iPSC-derived, complex, multilineage muscle models containing key isogenic cellular constituents of skeletal muscle, including vascular endothelial cells, pericytes, and motor neurons. These results lay the foundation for a human skeletal muscle organoid-like platform for disease modeling, regenerative medicine, and therapy development. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

High-resolution mapping of D16Led-1, Gart, Gas-r, Cbr, Pcp-4, and Erg on distal mouse chromosome 16

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mjaatvedt, A.E.; Citron, M.P.; Reeves, R.H.

1993-08-01

More than 500 backcross progeny from four inter-subspecific backcrosses were typed for six markers on distal mouse chromosome 16. Five of these represented genes that mapped within the Sod-1 to Ets-2 interval, which was shown previously to contain the weaver (wv) gene. The map order, including previously mapped reference markers, is (cen)-D16H21S16-D16Led-1-App-Sod-1-Gart-Gas-4-Cbr-wv-Pep-4-Erg-Ets-2. This gene order recapitulates the order of the genes on human chromosome 21 where known. Two of these markers further define the region containing the weaver gene to a 3.9-cM segment between Cbr and Pcp-4. In addition, Pep-4 was localized to human chromosome 21 by the presence ofmore » a human-specific restriction fragment in WAV-17, a mouse-human somatic cell hybrid with human chromosome 31 as the only human contribution. 26 refs., 3 figs., 3 tabs.« less

Acute exercise and physiological insulin induce distinct phosphorylation signatures on TBC1D1 and TBC1D4 proteins in human skeletal muscle

PubMed Central

Treebak, Jonas T; Pehmøller, Christian; Kristensen, Jonas M; Kjøbsted, Rasmus; Birk, Jesper B; Schjerling, Peter; Richter, Erik A; Goodyear, Laurie J; Wojtaszewski, Jørgen F P

2014-01-01

We investigated the phosphorylation signatures of two Rab-GTPase activating proteins TBC1D1 and TBC1D4 in human skeletal muscle in response to physical exercise and physiological insulin levels induced by a carbohydrate rich meal using a paired experimental design. Eight healthy male volunteers exercised in the fasted or fed state and muscle biopsies were taken before and immediately after exercise. We identified TBC1D1/4 phospho-sites that (1) did not respond to exercise or postprandial increase in insulin (TBC1D4: S666), (2) responded to insulin only (TBC1D4: S318), (3) responded to exercise only (TBC1D1: S237, S660, S700; TBC1D4: S588, S751), and (4) responded to both insulin and exercise (TBC1D1: T596; TBC1D4: S341, T642, S704). In the insulin-stimulated leg, Akt phosphorylation of both T308 and S473 correlated significantly with multiple sites on both TBC1D1 (T596) and TBC1D4 (S318, S341, S704). Interestingly, in the exercised leg in the fasted state TBC1D1 phosphorylation (S237, T596) correlated significantly with the activity of the α2/β2/γ3 AMPK trimer, whereas TBC1D4 phosphorylation (S341, S704) correlated with the activity of the α2/β2/γ1 AMPK trimer. Our data show differential phosphorylation of TBC1D1 and TBC1D4 in response to physiological stimuli in human skeletal muscle and support the idea that Akt and AMPK are upstream kinases. TBC1D1 phosphorylation signatures were comparable between in vitro contracted mouse skeletal muscle and exercised human muscle, and we show that AMPK regulated phosphorylation of these sites in mouse muscle. Contraction and exercise elicited a different phosphorylation pattern of TBC1D4 in mouse compared with human muscle, and although different circumstances in our experimental setup may contribute to this difference, the observation exemplifies that transferring findings between species is problematic. Key points Phosphorylation signature patterns on TBC1D1 and TBC1D4 proteins in the insulin–glucose pathway were investigated in human skeletal muscle in response to physiological insulin and exercise. In response to postprandial increase in insulin, Akt phosphorylation of T308 and S473 correlated significantly with sites on TBC1D1 (T596) and TBC1D4 (S318, S341, S704). Exercise induced phosphorylation of TBC1D1 (S237, T596) that correlated significantly with activity of the α2/β2/γ3 AMPK trimer, whereas TBC1D4 phosphorylation (S341, S704) with exercise correlated with activity of the α2/β2/γ1 AMPK trimer. TBC1D1 phosphorylation signatures with exercise/muscle contraction were comparable between human and mouse skeletal muscle, and AMPK regulated phosphorylation of these sites in mouse muscle, whereas contraction and exercise elicited different TBC1D4 phosphorylation patterns in mouse compared with human muscle. Our results show differential phosphorylation of TBC1D1 and TBC1D4 in response to physiological stimuli in human skeletal muscle and indicate that Akt and AMPK may be upstream kinases. PMID:24247980

Engineering the Controlled Assembly of Filamentous Injectisomes in E. coli K-12 for Protein Translocation into Mammalian Cells.

PubMed

Ruano-Gallego, David; Álvarez, Beatriz; Fernández, Luis Ángel

2015-09-18

Bacterial pathogens containing type III protein secretion systems (T3SS) assemble large needle-like protein complexes in the bacterial envelope, called injectisomes, for translocation of protein effectors into host cells. The application of these "molecular syringes" for the injection of proteins into mammalian cells is hindered by their structural and genomic complexity, requiring multiple polypeptides encoded along with effectors in various transcriptional units (TUs) with intricate regulation. In this work, we have rationally designed the controlled expression of the filamentous injectisomes found in enteropathogenic Escherichia coli (EPEC) in the nonpathogenic strain E. coli K-12. All structural components of EPEC injectisomes, encoded in a genomic island called the locus of enterocyte effacement (LEE), were engineered in five TUs (eLEEs) excluding effectors, promoters and transcriptional regulators. These eLEEs were placed under the control of the IPTG-inducible promoter Ptac and integrated into specific chromosomal sites of E. coli K-12 using a marker-less strategy. The resulting strain, named synthetic injector E. coli (SIEC), assembles filamentous injectisomes similar to those in EPEC. SIEC injectisomes form pores in the host plasma membrane and are able to translocate T3-substrate proteins (e.g., translocated intimin receptor, Tir) into the cytoplasm of HeLa cells reproducing the phenotypes of intimate attachment and polymerization of actin-pedestals elicited by EPEC bacteria. Hence, SIEC strain allows the controlled expression of functional filamentous injectisomes for efficient translocation of proteins with T3S-signals into mammalian cells.

Engineered strains of Streptococcus macedonicus towards an osmotic stress resistant phenotype retain their ability to produce the bacteriocin macedocin under hyperosmotic conditions.

PubMed

Anastasiou, Rania; Driessche, Gonzalez Van; Boutou, Effrossyni; Kazou, Maria; Alexandraki, Voula; Vorgias, Constantinos E; Devreese, Bart; Tsakalidou, Effie; Papadimitriou, Konstantinos

2015-10-20

Streptococcus macedonicus ACA-DC 198 produces the bacteriocin macedocin in milk only under low NaCl concentrations (<1.0%w/v). The thermosensitive plasmid pGh9:ISS1 was employed to generate osmotic stress resistant (osmr) mutants of S. macedonicus. Three osmr mutants showing integration of the vector in unique chromosomal sites were identified and the disrupted loci were characterized. Interestingly, the mutants were able to grow and to produce macedocin at considerably higher concentrations of NaCl compared to the wild-type (up to 4.0%w/v). The production of macedocin under hyperosmotic conditions solely by the osmr mutants was validated by the well diffusion assay and by mass spectrometry analysis. RT-PCR experiments demonstrated that the macedocin biosynthetic regulon was transcribed at high salt concentrations only in the mutants. Mutant osmr3, the most robust mutant, was converted in its markerless derivative (osmr3f). Co-culture of S. macedonicus with spores of Clostridium tyrobutyricum in milk demonstrated that only the osmr3f mutant and not the wild-type inhibited the growth of the spores under hyperosmotic conditions (i.e., 2.5%w/v NaCl) due to the production of macedocin. Our study shows how genetic manipulation of a strain towards a stress resistant phenotype could improve bacteriocin production under conditions of the same stress. Copyright © 2015 Elsevier B.V. All rights reserved.

Contrast sensitivity function in stereoscopic viewing of Gabor patches on a medical polarized three-dimensional stereoscopic display

NASA Astrophysics Data System (ADS)

Rousson, Johanna; Haar, Jérémy; Santal, Sarah; Kumcu, Asli; Platiša, Ljiljana; Piepers, Bastian; Kimpe, Tom; Philips, Wilfried

2016-03-01

While three-dimensional (3-D) imaging systems are entering hospitals, no study to date has explored the luminance calibration needs of 3-D stereoscopic diagnostic displays and if they differ from two-dimensional (2-D) displays. Since medical display calibration incorporates the human contrast sensitivity function (CSF), we first assessed the 2-D CSF for benchmarking and then examined the impact of two image parameters on the 3-D stereoscopic CSF: (1) five depth plane (DP) positions (between DP: -171 and DP: 2853 mm), and (2) three 3-D inclinations (0 deg, 45 deg, and 60 deg around the horizontal axis of a DP). Stimuli were stereoscopic images of a vertically oriented 2-D Gabor patch at one of seven frequencies ranging from 0.4 to 10 cycles/deg. CSFs were measured for seven to nine human observers with a staircase procedure. The results indicate that the 2-D CSF model remains valid for a 3-D stereoscopic display regardless of the amount of disparity between the stereo images. We also found that the 3-D CSF at DP≠0 does not differ from the 3-D CSF at DP=0 for DPs and disparities which allow effortless binocular fusion. Therefore, the existing 2-D medical luminance calibration algorithm remains an appropriate tool for calibrating polarized stereoscopic medical displays.

The Visible Human Project: From Body to Bits.

PubMed

Ackerman, Michael J

2017-01-01

Atlases of anatomy have long been a mainstay for visualizing and identifying features of the human body [1]. Many are constructed of idealized illustrations rendered so that structures are presented as three-dimensional (3-D) pictures. Others have employed photographs of actual dissections. Still others are composed of collections of artist renderings of organs or areas of interest. All rely on a basically two-dimensional (2-D) graphic display to depict and allow for a better understanding of a complicated 3-D structure.

Design and Fabrication of Human Skin by Three-Dimensional Bioprinting

PubMed Central

Lee, Vivian; Singh, Gurtej; Trasatti, John P.; Bjornsson, Chris; Xu, Xiawei; Tran, Thanh Nga; Yoo, Seung-Schik

2014-01-01

Three-dimensional (3D) bioprinting, a flexible automated on-demand platform for the free-form fabrication of complex living architectures, is a novel approach for the design and engineering of human organs and tissues. Here, we demonstrate the potential of 3D bioprinting for tissue engineering using human skin as a prototypical example. Keratinocytes and fibroblasts were used as constituent cells to represent the epidermis and dermis, and collagen was used to represent the dermal matrix of the skin. Preliminary studies were conducted to optimize printing parameters for maximum cell viability as well as for the optimization of cell densities in the epidermis and dermis to mimic physiologically relevant attributes of human skin. Printed 3D constructs were cultured in submerged media conditions followed by exposure of the epidermal layer to the air–liquid interface to promote maturation and stratification. Histology and immunofluorescence characterization demonstrated that 3D printed skin tissue was morphologically and biologically representative of in vivo human skin tissue. In comparison with traditional methods for skin engineering, 3D bioprinting offers several advantages in terms of shape- and form retention, flexibility, reproducibility, and high culture throughput. It has a broad range of applications in transdermal and topical formulation discovery, dermal toxicity studies, and in designing autologous grafts for wound healing. The proof-of-concept studies presented here can be further extended for enhancing the complexity of the skin model via the incorporation of secondary and adnexal structures or the inclusion of diseased cells to serve as a model for studying the pathophysiology of skin diseases. PMID:24188635

Design and fabrication of human skin by three-dimensional bioprinting.

PubMed

Lee, Vivian; Singh, Gurtej; Trasatti, John P; Bjornsson, Chris; Xu, Xiawei; Tran, Thanh Nga; Yoo, Seung-Schik; Dai, Guohao; Karande, Pankaj

2014-06-01

Three-dimensional (3D) bioprinting, a flexible automated on-demand platform for the free-form fabrication of complex living architectures, is a novel approach for the design and engineering of human organs and tissues. Here, we demonstrate the potential of 3D bioprinting for tissue engineering using human skin as a prototypical example. Keratinocytes and fibroblasts were used as constituent cells to represent the epidermis and dermis, and collagen was used to represent the dermal matrix of the skin. Preliminary studies were conducted to optimize printing parameters for maximum cell viability as well as for the optimization of cell densities in the epidermis and dermis to mimic physiologically relevant attributes of human skin. Printed 3D constructs were cultured in submerged media conditions followed by exposure of the epidermal layer to the air-liquid interface to promote maturation and stratification. Histology and immunofluorescence characterization demonstrated that 3D printed skin tissue was morphologically and biologically representative of in vivo human skin tissue. In comparison with traditional methods for skin engineering, 3D bioprinting offers several advantages in terms of shape- and form retention, flexibility, reproducibility, and high culture throughput. It has a broad range of applications in transdermal and topical formulation discovery, dermal toxicity studies, and in designing autologous grafts for wound healing. The proof-of-concept studies presented here can be further extended for enhancing the complexity of the skin model via the incorporation of secondary and adnexal structures or the inclusion of diseased cells to serve as a model for studying the pathophysiology of skin diseases.

Human lung fibroblast-derived matrix facilitates vascular morphogenesis in 3D environment and enhances skin wound healing.

PubMed

Du, Ping; Suhaeri, Muhammad; Ha, Sang Su; Oh, Seung Ja; Kim, Sang-Heon; Park, Kwideok

2017-05-01

Extracellular matrix (ECM) is crucial to many aspects of vascular morphogenesis and maintenance of vasculature function. Currently the recapitulation of angiogenic ECM microenvironment is still challenging, due mainly to its diverse components and complex organization. Here we investigate the angiogenic potential of human lung fibroblast-derived matrix (hFDM) in creating a three-dimensional (3D) vascular construct. hFDM was obtained via decellularization of in vitro cultured human lung fibroblasts and analyzed via immunofluorescence staining and ELISA, which detect multiple ECM macromolecules and angiogenic growth factors (GFs). Human umbilical vein endothelial cells (HUVECs) morphology was more elongated and better proliferative on hFDM than on gelatin-coated substrate. To prepare 3D construct, hFDM is collected, quantitatively analyzed, and incorporated in collagen hydrogel (Col) with HUVECs. Capillary-like structure (CLS) formation at 7day was significantly better with the groups containing higher doses of hFDM compared to the Col group (control). Moreover, the group (Col/hFDM/GFs) with both hFDM and angiogenic GFs (VEGF, bFGF, SDF-1) showed the synergistic activity on CLS formation and found much larger capillary lumen diameters with time. Further analysis of hFDM via angiogenesis antibody array kit reveals abundant biochemical cues, such as angiogenesis-related cytokines, GFs, and proteolytic enzymes. Significantly up-regulated expression of VE-cadherin and ECM-specific integrin subunits was also noticed in Col/hFDM/GFs. In addition, transplantation of Col/hFMD/GFs with HUVECs in skin wound model presents more effective re-epithelialization, many regenerated hair follicles, better transplanted cells viability, and advanced neovascularization. We believe that current system is a very promising platform for 3D vasculature construction in vitro and for cell delivery toward therapeutic applications in vivo. Functional 3D vasculature construction in vitro is still challenging due to the difficulty of recapitulating the complex angiogenic extracellular matrix (ECM) environment. Herein, we present a simple and practical method to create an angiogenic 3D environment via incorporation of human lung fibroblast-derived matrix (hFDM) into collagen hydrogel. We found that hFDM offers a significantly improved angiogenic microenvironment for HUVECs on 2D substrates and in 3D construct. A synergistic effect of hFDM and angiogenic growth factors has been well confirmed in 3D condition. The prevascularized 3D collagen constructs also facilitate skin wound healing. We believe that current system should be a convenient and powerful platform in engineering 3D vasculature in vitro, and in delivering cells for therapeutic purposes in vivo. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Individual Differences in Attentional Flexibility.

DTIC Science & Technology

1978-05-15

L A CADEMY ANNAPOLIS , MD 21*02 CDR PAUL NELSON NAVAL MEDICAL R& D COMMAND 1 Mr. Arnold I. Rubinstein CODE 1s14 Human Resoureces Program Manager...ARMY RESE AR CH iNSTITUTE Hea d Human Factors Engineering Div. 5001 EISENHOWER AVENUE Naval Air Development Center ALEXANDRI A , VA 22~~ 3 W~ rm inst...WEDNESDA Y, MAY 3, 1978 09:53 :0O—PDT PAGE 14 Army Air Force Dr. Joseph Ward 1 Air Force Human Resources Lab U.S. Army Research Institute AFHRL/PED 5001

Novel scalable 3D cell based model for in vitro neurotoxicity testing: Combining human differentiated neurospheres with gene expression and functional endpoints.

PubMed

Terrasso, Ana Paula; Pinto, Catarina; Serra, Margarida; Filipe, Augusto; Almeida, Susana; Ferreira, Ana Lúcia; Pedroso, Pedro; Brito, Catarina; Alves, Paula Marques

2015-07-10

There is an urgent need for new in vitro strategies to identify neurotoxic agents with speed, reliability and respect for animal welfare. Cell models should include distinct brain cell types and represent brain microenvironment to attain higher relevance. The main goal of this study was to develop and validate a human 3D neural model containing both neurons and glial cells, applicable for toxicity testing in high-throughput platforms. To achieve this, a scalable bioprocess for neural differentiation of human NTera2/cl.D1 cells in stirred culture systems was developed. Endpoints based on neuronal- and astrocytic-specific gene expression and functionality in 3D were implemented in multi-well format and used for toxicity assessment. The prototypical neurotoxicant acrylamide affected primarily neurons, impairing synaptic function; our results suggest that gene expression of the presynaptic marker synaptophysin can be used as sensitive endpoint. Chloramphenicol, described as neurotoxicant affected both cell types, with cytoskeleton markers' expression significantly reduced, particularly in astrocytes. In conclusion, a scalable and reproducible process for production of differentiated neurospheres enriched in mature neurons and functional astrocytes was obtained. This 3D approach allowed efficient production of large numbers of human differentiated neurospheres, which in combination with gene expression and functional endpoints are a powerful cell model to evaluate human neuronal and astrocytic toxicity. Copyright © 2014 Elsevier B.V. All rights reserved.

Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols

PubMed Central

Jeziorowska, Dorota; Fontaine, Vincent; Jouve, Charlène; Villard, Eric; Dussaud, Sébastien; Akbar, David; Letang, Valérie; Cervello, Pauline; Itier, Jean-Michiel; Pruniaux, Marie-Pierre; Hulot, Jean-Sébastien

2017-01-01

Human induced pluripotent stem cells (iPSCs) represent a powerful human model to study cardiac disease in vitro, notably channelopathies and sarcomeric cardiomyopathies. Different protocols for cardiac differentiation of iPSCs have been proposed either based on embroid body formation (3D) or, more recently, on monolayer culture (2D). We performed a direct comparison of the characteristics of the derived cardiomyocytes (iPSC-CMs) on day 27 ± 2 of differentiation between 3D and 2D differentiation protocols with two different Wnt-inhibitors were compared: IWR1 (inhibitor of Wnt response) or IWP2 (inhibitor of Wnt production). We firstly found that the level of Troponin T (TNNT2) expression measured by FACS was significantly higher for both 2D protocols as compared to the 3D protocol. In the three methods, iPSC-CM show sarcomeric structures. However, iPSC-CM generated in 2D protocols constantly displayed larger sarcomere lengths as compared to the 3D protocol. In addition, mRNA and protein analyses reveal higher cTNi to ssTNi ratios in the 2D protocol using IWP2 as compared to both other protocols, indicating a higher sarcomeric maturation. Differentiation of cardiac myocytes with 2D monolayer-based protocols and the use of IWP2 allows the production of higher yield of cardiac myocytes that have more suitable characteristics to study sarcomeric cardiomyopathies. PMID:28587156

The HIV-1 viral protein Tat increases glutamate and decreases GABA exocytosis from human and mouse neocortical nerve endings.

PubMed

Musante, Veronica; Summa, Maria; Neri, Elisa; Puliti, Aldamaria; Godowicz, Tomasz T; Severi, Paolo; Battaglia, Giuseppe; Raiteri, Maurizio; Pittaluga, Anna

2010-08-01

Human immunodeficiency virus-1 (HIV-1)-encoded transactivator of transcription (Tat) potentiated the depolarization-evoked exocytosis of [(3)H]D-aspartate ([(3)H]D-ASP) from human neocortical terminals. The metabotropic glutamate (mGlu) 1 receptor antagonist 7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt) prevented this effect, whereas the mGlu5 receptor antagonist 2-methyl-6-(phenylethynyl) pyridine hydrochloride (MPEP) was ineffective. Western blot analysis showed that human neocortex synaptosomes possess mGlu1 and mGlu5 receptors. Tat potentiated the K(+)-evoked release of [(3)H]D-ASP or of endogenous glutamate from mouse neocortical synaptosomes in a CPCCOEt-sensitive and MPEP-insensitive manner. Deletion of mGlu1 receptors (crv4/crv4 mice) or mGlu5 receptors (mGlu5(-/-)mouse) silenced Tat effects. Tat enhanced inositol 1,4,5-trisphosphate production in human and mouse neocortical synaptosomes, consistent with the involvement of group I mGlu receptors. Tat inhibited the K(+)-evoked release of [(3)H]gamma-aminobutyric acid ([(3)H]GABA) from human synaptosomes and that of endogenous GABA or [(3)H]GABA from mouse nerve terminals; the inhibition was insensitive to CPCCOEt or MPEP. Tat-induced effects were retained by Tat(37-72) but not by Tat(48-85). In mouse neocortical slices, Tat facilitated the K(+)- and the veratridine-induced release of [(3)H]D-ASP in a CPCCOEt-sensitive manner and was ineffective in crv4/crv4 mouse slices. These observations are relevant to the comprehension of the pathophysiological effects of Tat in central nervous system and may suggest new potential therapeutic approaches to the cure of HIV-1-associated dementia.

Identification of stable reference genes for gene expression analysis of three-dimensional cultivated human bone marrow-derived mesenchymal stromal cells for bone tissue engineering.

PubMed

Rauh, Juliane; Jacobi, Angela; Stiehler, Maik

2015-02-01

The principles of tissue engineering (TE) are widely used for bone regeneration concepts. Three-dimensional (3D) cultivation of autologous human mesenchymal stromal cells (MSCs) on porous scaffolds is the basic prerequisite to generate newly formed bone tissue. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a specific and sensitive analytical tool for the measurement of mRNA-levels in cells or tissues. For an accurate quantification of gene expression levels, stably expressed reference genes (RGs) are essential to obtain reliable results. Since the 3D environment can affect a cell's morphology, proliferation, and gene expression profile compared with two-dimensional (2D) cultivation, there is a need to identify robust RGs for the quantification of gene expression. So far, this issue has not been adequately investigated. The aim of this study was to identify the most stably expressed RGs for gene expression analysis of 3D-cultivated human bone marrow-derived MSCs (BM-MSCs). For this, we analyzed the gene expression levels of n=31 RGs in 3D-cultivated human BM-MSCs from six different donors compared with conventional 2D cultivation using qRT-PCR. MSCs isolated from bone marrow aspirates were cultivated on human cancellous bone cube scaffolds for 14 days. Osteogenic differentiation was assessed by cell-specific alkaline phosphatase (ALP) activity and expression of osteogenic marker genes. Expression levels of potential reference and target genes were quantified using commercially available TaqMan(®) assays. mRNA expression stability of RGs was determined by calculating the coefficient of variation (CV) and using the algorithms of geNorm and NormFinder. Using both algorithms, we identified TATA box binding protein (TBP), transferrin receptor (p90, CD71) (TFRC), and hypoxanthine phosphoribosyltransferase 1 (HPRT1) as the most stably expressed RGs in 3D-cultivated BM-MSCs. Notably, genes that are routinely used as RGs, for example, beta actin (ACTB) and ribosomal protein L37a (RPL37A), were among the least stable genes. We recommend the combined use of TBP, TFRC, and HPRT1 for the accurate and robust normalization of qRT-PCR data of 3D-cultivated human BM-MSCs.

Identification of Stable Reference Genes for Gene Expression Analysis of Three-Dimensional Cultivated Human Bone Marrow-Derived Mesenchymal Stromal Cells for Bone Tissue Engineering

PubMed Central

Rauh, Juliane; Jacobi, Angela

2015-01-01

The principles of tissue engineering (TE) are widely used for bone regeneration concepts. Three-dimensional (3D) cultivation of autologous human mesenchymal stromal cells (MSCs) on porous scaffolds is the basic prerequisite to generate newly formed bone tissue. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a specific and sensitive analytical tool for the measurement of mRNA-levels in cells or tissues. For an accurate quantification of gene expression levels, stably expressed reference genes (RGs) are essential to obtain reliable results. Since the 3D environment can affect a cell's morphology, proliferation, and gene expression profile compared with two-dimensional (2D) cultivation, there is a need to identify robust RGs for the quantification of gene expression. So far, this issue has not been adequately investigated. The aim of this study was to identify the most stably expressed RGs for gene expression analysis of 3D-cultivated human bone marrow-derived MSCs (BM-MSCs). For this, we analyzed the gene expression levels of n=31 RGs in 3D-cultivated human BM-MSCs from six different donors compared with conventional 2D cultivation using qRT-PCR. MSCs isolated from bone marrow aspirates were cultivated on human cancellous bone cube scaffolds for 14 days. Osteogenic differentiation was assessed by cell-specific alkaline phosphatase (ALP) activity and expression of osteogenic marker genes. Expression levels of potential reference and target genes were quantified using commercially available TaqMan® assays. mRNA expression stability of RGs was determined by calculating the coefficient of variation (CV) and using the algorithms of geNorm and NormFinder. Using both algorithms, we identified TATA box binding protein (TBP), transferrin receptor (p90, CD71) (TFRC), and hypoxanthine phosphoribosyltransferase 1 (HPRT1) as the most stably expressed RGs in 3D-cultivated BM-MSCs. Notably, genes that are routinely used as RGs, for example, beta actin (ACTB) and ribosomal protein L37a (RPL37A), were among the least stable genes. We recommend the combined use of TBP, TFRC, and HPRT1 for the accurate and robust normalization of qRT-PCR data of 3D-cultivated human BM-MSCs. PMID:25000821

Prevascularization of 3D printed bone scaffolds by bioactive hydrogels and cell co-culture.

PubMed

Kuss, Mitchell A; Wu, Shaohua; Wang, Ying; Untrauer, Jason B; Li, Wenlong; Lim, Jung Yul; Duan, Bin

2017-09-13

Vascularization is a fundamental prerequisite for large bone construct development and remains one of the main challenges of bone tissue engineering. Our current study presents the combination of 3D printing technique with a hydrogel-based prevascularization strategy to generate prevascularized bone constructs. Human adipose derived mesenchymal stem cells (ADMSC) and human umbilical vein endothelial cells (HUVEC) were encapsulated within our bioactive hydrogels, and the effects of culture conditions on in vitro vascularization were determined. We further generated composite constructs by forming 3D printed polycaprolactone/hydroxyapatite scaffolds coated with cell-laden hydrogels and determined how the co-culture affected vascularization and osteogenesis. It was demonstrated that 3D co-cultured ADMSC-HUVEC generated capillary-like networks within the porous 3D printed scaffold. The co-culture systems promoted in vitro vascularization, but had no significant effects on osteogenesis. The prevascularized constructs were subcutaneously implanted into nude mice to evaluate the in vivo vascularization capacity and the functionality of engineered vessels. The hydrogel systems facilitated microvessel and lumen formation and promoted anastomosis of vascular networks of human origin with host murine vasculature. These findings demonstrate the potential of prevascularized 3D printed scaffolds with anatomical shape for the healing of larger bone defects. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

Recognition of complex human behaviours using 3D imaging for intelligent surveillance applications

NASA Astrophysics Data System (ADS)

Yao, Bo; Lepley, Jason J.; Peall, Robert; Butler, Michael; Hagras, Hani

2016-10-01

We introduce a system that exploits 3-D imaging technology as an enabler for the robust recognition of the human form. We combine this with pose and feature recognition capabilities from which we can recognise high-level human behaviours. We propose a hierarchical methodology for the recognition of complex human behaviours, based on the identification of a set of atomic behaviours, individual and sequential poses (e.g. standing, sitting, walking, drinking and eating) that provides a framework from which we adopt time-based machine learning techniques to recognise complex behaviour patterns.

[The virtual reality simulation research of China Mechanical Virtual Human based on the Creator/Vega].

PubMed

Wei, Gaofeng; Tang, Gang; Fu, Zengliang; Sun, Qiuming; Tian, Feng

2010-10-01

The China Mechanical Virtual Human (CMVH) is a human musculoskeletal biomechanical simulation platform based on China Visible Human slice images; it has great realistic application significance. In this paper is introduced the construction method of CMVH 3D models. Then a simulation system solution based on Creator/Vega is put forward for the complex and gigantic data characteristics of the 3D models. At last, combined with MFC technology, the CMVH simulation system is developed and a running simulation scene is given. This paper provides a new way for the virtual reality application of CMVH.

Development and Assessment of a New 3D Neuroanatomy Teaching Tool for MRI Training

ERIC Educational Resources Information Center

Drapkin, Zachary A.; Lindgren, Kristen A.; Lopez, Michael J.; Stabio, Maureen E.

2015-01-01

A computerized three-dimensional (3D) neuroanatomy teaching tool was developed for training medical students to identify subcortical structures on a magnetic resonance imaging (MRI) series of the human brain. This program allows the user to transition rapidly between two-dimensional (2D) MRI slices, 3D object composites, and a combined model in…

Extraction of the 3D local orientation of myocytes in human cardiac tissue using X-ray phase-contrast micro-tomography and multi-scale analysis.

PubMed

Varray, François; Mirea, Iulia; Langer, Max; Peyrin, Françoise; Fanton, Laurent; Magnin, Isabelle E

2017-05-01

This paper presents a methodology to access the 3D local myocyte arrangements in fresh human post-mortem heart samples. We investigated the cardiac micro-structure at a high and isotropic resolution of 3.5 µm in three dimensions using X-ray phase micro-tomography at the European Synchrotron Radiation Facility. We then processed the reconstructed volumes to extract the 3D local orientation of the myocytes using a multi-scale approach with no segmentation. We created a simplified 3D model of tissue sample made of simulated myocytes with known size and orientations, to evaluate our orientation extraction method. Afterwards, we applied it to 2D histological cuts and to eight 3D left ventricular (LV) cardiac tissue samples. Then, the variation of the helix angles, from the endocardium to the epicardium, was computed at several spatial resolutions ranging from 3.6 3  mm 3 to 112 3  µm 3 . We measure an increased range of 20° to 30° from the coarsest resolution level to the finest level in the experimental samples. This result is in line with the higher values measured from histology. The displayed tractography demonstrates a rather smooth evolution of the transmural helix angle in six LV samples and a sudden discontinuity of the helix angle in two septum samples. These measurements bring a new vision of the human heart architecture from macro- to micro-scale. Copyright © 2017 Elsevier B.V. All rights reserved.

How 3D immersive visualization is changing medical diagnostics

NASA Astrophysics Data System (ADS)

Koning, Anton H. J.

2011-03-01

Originally the only way to look inside the human body without opening it up was by means of two dimensional (2D) images obtained using X-ray equipment. The fact that human anatomy is inherently three dimensional leads to ambiguities in interpretation and problems of occlusion. Three dimensional (3D) imaging modalities such as CT, MRI and 3D ultrasound remove these drawbacks and are now part of routine medical care. While most hospitals 'have gone digital', meaning that the images are no longer printed on film, they are still being viewed on 2D screens. However, this way valuable depth information is lost, and some interactions become unnecessarily complex or even unfeasible. Using a virtual reality (VR) system to present volumetric data means that depth information is presented to the viewer and 3D interaction is made possible. At the Erasmus MC we have developed V-Scope, an immersive volume visualization system for visualizing a variety of (bio-)medical volumetric datasets, ranging from 3D ultrasound, via CT and MRI, to confocal microscopy, OPT and 3D electron-microscopy data. In this talk we will address the advantages of such a system for both medical diagnostics as well as for (bio)medical research.

Using videogrammetry and 3D image reconstruction to identify crime suspects

NASA Astrophysics Data System (ADS)

Klasen, Lena M.; Fahlander, Olov

1997-02-01

The anthropometry and movements are unique for every individual human being. We identify persons we know by recognizing the way the look and move. By quantifying these measures and using image processing methods this method can serve as a tool in the work of the police as a complement to the ability of the human eye. The idea is to use virtual 3-D parameterized models of the human body to measure the anthropometry and movements of a crime suspect. The Swedish National Laboratory of Forensic Science in cooperation with SAAB Military Aircraft have developed methods for measuring the lengths of persons from video sequences. However, there is so much unused information in a digital image sequence from a crime scene. The main approach for this paper is to give an overview of the current research project at Linkoping University, Image Coding Group where methods to measure anthropometrical data and movements by using virtual 3-D parameterized models of the person in the crime scene are being developed. The length of an individual might vary up to plus or minus 10 cm depending on whether the person is in upright position or not. When measuring during the best available conditions, the length still varies within plus or minus 1 cm. Using a full 3-D model provides a rich set of anthropometric measures describing the person in the crime scene. Once having obtained such a model the movements can be quantified as well. The results depend strongly on the accuracy of the 3-D model and the strategy of having such an accurate 3-D model is to make one estimate per image frame by using 3-D scene reconstruction, and an averaged 3-D model as the final result from which the anthropometry and movements are calculated.

The study of early human embryos using interactive 3-dimensional computer reconstructions.

PubMed

Scarborough, J; Aiton, J F; McLachlan, J C; Smart, S D; Whiten, S C

1997-07-01

Tracings of serial histological sections from 4 human embryos at different Carnegie stages were used to create 3-dimensional (3D) computer models of the developing heart. The models were constructed using commercially available software developed for graphic design and the production of computer generated virtual reality environments. They are available as interactive objects which can be downloaded via the World Wide Web. This simple method of 3D reconstruction offers significant advantages for understanding important events in morphological sciences.

Selected Issues in DoD’s Radio Frequency Identification (RFID) Implementation

DTIC Science & Technology

2006-04-01

Evaluation of human exposure to electromagnetic fields from devices operating in the frequency range 0 Hz to 10 GHz, used in Electronic...standard for human exposure to RF Signal, 3 kHz-300 GHz BS EN 50364 Limitation of human exposure to electromagnetic fields from devices operating in the...Management and DoD Explosives Safety Board, and DoDD 6055.9-STD, DoD Ammunition and Explosives Safety Standards. Exposure of people to electromagnetic

A model of the complex between human {beta}-microseminoprotein and CRISP-3 based on NMR data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ghasriani, Houman; Fernlund, Per; Udby, Lene

2009-01-09

{beta}-Microseminoprotein (MSP), a 10 kDa seminal plasma protein, forms a tight complex with cysteine-rich secretory protein 3 (CRISP-3) from granulocytes. The 3D structure of human MSP has been determined but there is as yet no 3D structure for CRISP-3. We have now studied the complex between human MSP and CRISP-3 with multidimensional NMR. {sup 15}N-HSQC spectra show substantial differences between free and complexed hMSP. Using several 3D-NMR spectra of triply labeled hMSP in complex with a recombinant N-terminal domain of CRISP-3, most of the backbone of hMSP could be assigned. The data show that only one side of hMSP, comprisingmore » {beta}-strands 1, 4, 5, and 8 are affected by the complex formation, indicating that {beta}-strands 1 and 8 form the main binding surface. Based on this we present a tentative structure for the hMSP-CRISP-3 complex using the known crystal structure of triflin as a model of CRISP-3.« less

Generation of C3- and C2-deuterated L-lactic acid by human erythrocytes exposed to D-[1-13C]glucose, D-[2-13C]glucose and D-[6-13C]glucose in the presence of D2O.

PubMed

Malaisse, W J; Biesemans, M; Willem, R

1994-05-01

1. The generation of C2- and C3-deuterated L-lactate was monitored by 13C NMR in human erythrocytes exposed to D-[1-13C]glucose, D-[2-13C]glucose or D-[6-13C]glucose and incubated in a medium prepared in D2O. 2. The results suggested that the deuteration of the C1 of D-fructose 6-phosphate in the phosphoglucoisomerase reaction, the deuteration of the C1 of D-glyceraldehyde-3-phosphate in the sequence of reactions catalyzed by triose phosphate isomerase and aldolase and the deuteration of the C3 of pyruvate in the reaction catalyzed by pyruvate kinase were all lower than expected from equilibration with D2O. 3. Moreover, about 40% of the molecules of pyruvate generated by glycolysis apparently underwent deuteration on their C3 during interconversion of the 2-keto acid and L-alanine in the reaction catalyzed by glutamate-pyruvate transaminase. 4. The occurrence of the latter process was also documented in cells exposed to exogenous [3-13C]pyruvate. 5. This methodological approach is proposed to provide a new tool to assess in intact cells the extent of back-and-forth interconversion of selected metabolic intermediates.

Enhancement of hepatic 4-hydroxylation of 25-hydroxyvitamin D3 through CYP3A4 induction in vitro and in vivo: implications for drug-induced osteomalacia.

PubMed

Wang, Zhican; Lin, Yvonne S; Dickmann, Leslie J; Poulton, Emma-Jane; Eaton, David L; Lampe, Johanna W; Shen, Danny D; Davis, Connie L; Shuhart, Margaret C; Thummel, Kenneth E

2013-05-01

Long-term therapy with certain drugs, especially cytochrome P450 (P450; CYP)-inducing agents, confers an increased risk of osteomalacia that is attributed to vitamin D deficiency. Human CYP24A1, CYP3A4, and CYP27B1 catalyze the inactivation and activation of vitamin D and have been implicated in the adverse drug response. In this study, the inducibility of these enzymes and monohydroxylation of 25-hydroxyvitamin D3 (25OHD3) were evaluated after exposure to P450-inducing drugs. With human hepatocytes, treatment with phenobarbital, hyperforin, carbamazepine, and rifampin significantly increased the levels of CYP3A4, but not CYP24A1 or CYP27B1 mRNA. In addition, rifampin pretreatment resulted in an 8-fold increase in formation of the major metabolite of 25OHD3, 4β,25(OH)2D3. This inductive effect was blocked by the addition of 6',7'-dihydroxybergamottin, a selective CYP3A4 inhibitor. With human renal proximal tubular HK-2 cells, treatment with the same inducers did not alter CYP3A4, CYP24A1, or CYP27B1 expression. 24R,25(OH)2 D3 was the predominant monohydroxy metabolite produced from 25OHD3, but its formation was unaffected by the inducers. With healthy volunteers, the mean plasma concentration of 4β,25(OH)2D3 was increased 60% (p < 0.01) after short-term rifampin administration. This was accompanied by a statistically significant reduction in plasma 1α,25(OH)2D3 (-10%; p = 0.03), and a nonsignificant change in 24R,25(OH)2D3 (-8%; p = 0.09) levels. Further analysis revealed a negative correlation between the increase in 4β,25(OH)2D3 and decrease in 1α,25(OH)2D3 levels. Examination of the plasma monohydroxy metabolite/25OHD3 ratios indicated selective induction of the CYP3A4-dependent 4β-hydroxylation pathway of 25OHD3 elimination. These results suggest that induction of hepatic CYP3A4 may be important in the etiology of drug-induced osteomalacia. Copyright © 2013 American Society for Bone and Mineral Research.

Evaluation of 3D printing materials for fabrication of a novel multi-functional 3D thyroid phantom for medical dosimetry and image quality

NASA Astrophysics Data System (ADS)

Alssabbagh, Moayyad; Tajuddin, Abd Aziz; Abdulmanap, Mahayuddin; Zainon, Rafidah

2017-06-01

Recently, the three-dimensional printer has started to be utilized strongly in medical industries. In the human body, many parts or organs can be printed from 3D images to meet accurate organ geometries. In this study, five common 3D printing materials were evaluated in terms of their elementary composition and the mass attenuation coefficients. The online version of XCOM photon cross-section database was used to obtain the attenuation values of each material. The results were compared with the attenuation values of the thyroid listed in the International Commission on Radiation Units and Measurements - ICRU 44. Two original thyroid models (hollow-inside and solid-inside) were designed from scratch to be used in nuclear medicine, diagnostic radiology and radiotherapy for dosimetry and image quality purposes. Both designs have three holes for installation of radiation dosimeters. The hollow-inside model has more two holes in the top for injection the radioactive materials. The attenuation properties of the Polylactic Acid (PLA) material showed a very good match with the thyroid tissue, which it was selected to 3D print the phantom using open source RepRap, Prusa i3 3D printer. The scintigraphy images show that the phantom simulates a real healthy thyroid gland and thus it can be used for image quality purposes. The measured CT numbers of the PA material after the 3D printing show a close match with the human thyroid CT numbers. Furthermore, the phantom shows a good accommodation of the TLD dosimeters inside the holes. The 3D fabricated thyroid phantom simulates the real shape of the human thyroid gland with a changeable geometrical shape-size feature to fit different age groups. By using 3D printing technology, the time required to fabricate the 3D phantom was considerably shortened compared to the longer conventional methods, where it took only 30 min to print out the model. The 3D printing material used in this study is commercially available and cost-effective compared to current commercial tissue-equivalent materials.

Technical Report on the Modification of 3-Dimensional Non-contact Human Body Laser Scanner for the Measurement of Anthropometric Dimensions: Verification of its Accuracy and Precision.

PubMed

Jafari Roodbandi, Akram Sadat; Naderi, Hamid; Hashenmi-Nejad, Naser; Choobineh, Alireza; Baneshi, Mohammad Reza; Feyzi, Vafa

2017-01-01

Introduction: Three-dimensional (3D) scanners are widely used in medicine. One of the applications of 3D scanners is the acquisition of anthropometric dimensions for ergonomics and the creation of an anthropometry data bank. The aim of this study was to evaluate the precision and accuracy of a modified 3D scanner fabricated in this study. Methods: In this work, a 3D scan of the human body was obtained using DAVID Laser Scanner software and its calibration background, a linear low-power laser, and one advanced webcam. After the 3D scans were imported to the Geomagic software, 10 anthropometric dimensions of 10 subjects were obtained. The measurements of the 3D scanner were compared to the measurements of the same dimensions by a direct anthropometric method. The precision and accuracy of the measurements of the 3D scanner were then evaluated. The obtained data were analyzed using an independent sample t test with the SPSS software. Results: The minimum and maximum measurement differences from three consecutive scans by the 3D scanner were 0.03 mm and 18 mm, respectively. The differences between the measurements by the direct anthropometry method and the 3D scanner were not statistically significant. Therefore, the accuracy of the 3D scanner is acceptable. Conclusion: Future studies will need to focus on the improvement of the scanning speed and the quality of the scanned image.

Technical Report on the Modification of 3-Dimensional Non-contact Human Body Laser Scanner for the Measurement of Anthropometric Dimensions: Verification of its Accuracy and Precision

PubMed Central

Jafari Roodbandi, Akram Sadat; Naderi, Hamid; Hashenmi-Nejad, Naser; Choobineh, Alireza; Baneshi, Mohammad Reza; Feyzi, Vafa

2017-01-01

Introduction: Three-dimensional (3D) scanners are widely used in medicine. One of the applications of 3D scanners is the acquisition of anthropometric dimensions for ergonomics and the creation of an anthropometry data bank. The aim of this study was to evaluate the precision and accuracy of a modified 3D scanner fabricated in this study. Methods: In this work, a 3D scan of the human body was obtained using DAVID Laser Scanner software and its calibration background, a linear low-power laser, and one advanced webcam. After the 3D scans were imported to the Geomagic software, 10 anthropometric dimensions of 10 subjects were obtained. The measurements of the 3D scanner were compared to the measurements of the same dimensions by a direct anthropometric method. The precision and accuracy of the measurements of the 3D scanner were then evaluated. The obtained data were analyzed using an independent sample t test with the SPSS software. Results: The minimum and maximum measurement differences from three consecutive scans by the 3D scanner were 0.03 mm and 18 mm, respectively. The differences between the measurements by the direct anthropometry method and the 3D scanner were not statistically significant. Therefore, the accuracy of the 3D scanner is acceptable. Conclusion: Future studies will need to focus on the improvement of the scanning speed and the quality of the scanned image. PMID:28912940

In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor.

PubMed

Mueller, Daniel; Tascher, Georg; Müller-Vieira, Ursula; Knobeloch, Daniel; Nuessler, Andreas K; Zeilinger, Katrin; Heinzle, Elmar; Noor, Fozia

2011-08-01

As the major research focus is shifting to three-dimensional (3D) cultivation techniques, hollow-fiber bioreactors, allowing the formation of tissue-like structures, show immense potential as they permit controlled in vitro cultivation while supporting the in vivo environment. In this study we carried out a systematic and detailed physiological characterization of human liver cells in a 3D hollow-fiber bioreactor system continuously run for > 2 weeks. Primary human hepatocytes were maintained viable and functional over the whole period of cultivation. Both general cellular functions, e.g. oxygen uptake, amino acid metabolism and substrate consumption, and liver-specific functions, such as drug-metabolizing capacities and the production of liver-specific metabolites were found to be stable for > 2 weeks. As expected, donor-to-donor variability was observed in liver-specific functions, namely urea and albumin production. Moreover, we show the maintenance of primary human hepatocytes in serum-free conditions in this set-up. The stable basal cytochrome P450 activity 3 weeks after isolation of the cells demonstrates the potential of such a system for pharmacological applications. Liver cells in the presented 3D bioreactor system could eventually be used not only for long-term metabolic and toxicity studies but also for chronic repeated dose toxicity assessment. Copyright © 2011 John Wiley & Sons, Ltd.

Human Parotid Gland Alpha-Amylase Secretion as a Function of Chronic Hyperbaric Exposure

DTIC Science & Technology

1979-01-01

parotid ...Pullman, WA 99163 Gilman, S. C, G. J. Fischer, R. J. Biersner, R. D. Thornton, and D. A. Miller. 1979. Human parotid gland alpha-amylase secretion...as a function of chronic hyperbaric exposure. Undersea Biomed. Res. 6(3):303-307.—Secretion of a-amylase by the human parotid gland increased

Use of cues in virtual reality depends on visual feedback.

PubMed

Fulvio, Jacqueline M; Rokers, Bas

2017-11-22

3D motion perception is of central importance to daily life. However, when tested in laboratory settings, sensitivity to 3D motion signals is found to be poor, leading to the view that heuristics and prior assumptions are critical for 3D motion perception. Here we explore an alternative: sensitivity to 3D motion signals is context-dependent and must be learned based on explicit visual feedback in novel environments. The need for action-contingent visual feedback is well-established in the developmental literature. For example, young kittens that are passively moved through an environment, but unable to move through it themselves, fail to develop accurate depth perception. We find that these principles also obtain in adult human perception. Observers that do not experience visual consequences of their actions fail to develop accurate 3D motion perception in a virtual reality environment, even after prolonged exposure. By contrast, observers that experience the consequences of their actions improve performance based on available sensory cues to 3D motion. Specifically, we find that observers learn to exploit the small motion parallax cues provided by head jitter. Our findings advance understanding of human 3D motion processing and form a foundation for future study of perception in virtual and natural 3D environments.