Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells.

BACKGROUND: Diabetes is a disease affecting over 500 million people globally due to insulin insufficiency or insensitivity. For individuals with type 1 diabetes, pancreatic islet transplantation can help regulate their blood glucose levels. However, the scarcity of cadaveric donor islets limits the number of people that could receive this therapy. To address this issue, human pluripotent stem cells offer a potentially unlimited source for generating insulin-producing cells through directed differentiation. Several protocols have been developed to make stem cell-derived insulin-producing cells. However, there is a lack of knowledge regarding the bioprocess parameters associated with these differentiation protocols and how they can be utilized to increase the cell yield. METHODS: We investigated various bioprocess parameters and quality target product profiles that may influence the differentiation pipeline using a seven-stage protocol in a scalable manner with CellSTACKs and vertical wheel bioreactors (PBS-Minis). RESULTS: Cells maintained > 80% viability through all stages of differentiation and appropriately expressed stage-specific markers. During the initial four stages leading up to the development of pancreatic progenitors, there was an increase in cell numbers. Following pancreatic progenitor stage, there was a gradual decrease in the percentage of proliferative cells, as determined by Ki67 positivity, and a significant loss of cells during the period of endocrine differentiation. By minimizing the occurrence of aggregate fusion, we were able to enhance cell yield during the later stages of differentiation. We suggest that glucose utilization and lactate production are cell quality attributes that should be considered during the characterization of insulin-producing cells derived from stem cells. Our findings also revealed a gradual metabolic shift from glycolysis, during the initial four stages of pancreatic progenitor formation, to oxidative phosphorylation later on during endocrine differentiation. Furthermore, the resulting insulin-producing cells exhibited a response to several secretagogues, including high glucose. CONCLUSION: This study demonstrates process parameters such as glucose consumption and lactate production rates that may be used to facilitate the scalable manufacture of stem cell-derived insulin-producing cells.

Validation of a videogrammetry technique for analysing American football helmet kinematics.

Professional American football games are recorded in digital video with multiple cameras, often at high resolution and high frame rates. The purpose of this study was to evaluate the accuracy of a videogrammetry technique to calculate translational and rotational helmet velocity before, during and after a helmet impact. In total, 10 football impacts were staged in a National Football League (NFL) stadium by propelling helmeted 50th percentile male crash test dummies into each other or the ground at speeds and orientations representative of concussive impacts for NFL players. The tests were recorded by experienced sports film crews to obtain video coverage and quality typically available for NFL games. A videogrammetry procedure was used to track the position and rotation of the helmet throughout the relevant time interval of the head impact. Compared with rigidly mounted retroreflective marker three dimensional (3-D) motion tracking that was concurrently collected in the experiments, videogrammetry accurately calculated changes in translational and rotational velocity of the helmet using high frame rate (two cameras at 240 Hz) video (7% and 15% error, respectively). Low frame rate (2 cameras at 60 Hz) video was adequate for calculating pre-impact translational velocity but not for calculating the translational or rotational velocity change of the helmet during impact.

The biomechanics of concussive helmet-to-ground impacts in the National Football league.

This paper presents a detailed characterization of helmet-to-ground impacts in the National Football League. Video analysis was performed for 16 head-to-ground impacts that caused concussions. Average resultant closing velocity was 8.3 m/s at an angle nearly 45° to the surface. Preimpact rotational velocity of the helmet ranged from negligible to as high as 54.1 rad/s. Helmet impacts were concentrated on the posterior and lateral aspects. To study the interaction in greater detail, a helmeted anthropomorphic test device (ATD) was launched over a football field and fell to the ground in various impact conditions. Substantial decoupling between the helmet and the head was observed, such that the head rebounded within the helmet and underwent changes in linear and rotational motion greater than those of the helmet. Vertical helmet rebound was also observed; the helmet underwent a change in vertical velocity on average 24% greater than the vertical component of its closing velocity. Frictional interaction between the helmet and the ground surface caused the helmet to undergo an average horizontal change in velocity of 57% of the horizontal component of its closing velocity. Finally, the duration of a helmet-to-ground impact was generally in the range of 15 - 30 ms, suggesting that the impact surface provides little ride-down. Lengthening this duration could be beneficial both by reducing the peak linear and rotational acceleration and by shifting the impact toward a time regime where brain strain is related to rotational acceleration rather than rotational velocity.

Relative Motion Between the Helmet and the Head in Football Impact Test.

Approximately 1.6-3.8 million sports-related traumatic brain injuries occur each year in the U.S. Researchers track the head motion using a variety of techniques to study the head injury biomechanics. To understand how helmets provide head protection, quantification of the relative motion between the head and the helmet is necessary. The purpose of this study was to compare helmet and head kinematics and quantify the relative motion of helmet with respect to head during experimental representations of on-field American football impact scenarios. Seven helmet-to-helmet impact configurations were simulated by propelling helmeted crash test dummies into each other. Head and helmet kinematics were measured with instrumentation and an optical motion capture system. The analysis of results, from 10 ms prior to the helmet contact to 20 ms after the loss of helmet contact, showed that the helmets translated 12-41 mm and rotated up to 37 deg with respect to the head. The peak resultant linear acceleration of the helmet was about 2-5 times higher than the head. The peak resultant angular velocity of the helmet ranged from 37% less to 71% more than the head, depending on the impact conditions. The results of this study demonstrate that the kinematics of the head and the helmet are noticeably different and that the helmet rotates significantly with respect to the head during impacts. Therefore, capturing the helmet kinematics using a video motion tracking methodology is not sufficient to study the biomechanics of the head. Head motion must be measured independently of the helmet.

The Athletic Shoe in Football.

BACKGROUND: Foot and ankle injuries are common in sports, particularly in cleated athletes. Traditionally, the athletic shoe has not been regarded as a piece of protective equipment but rather as a part of the uniform, with a primary focus on performance and subjective feedback measures of comfort. Changes in turf and shoe design have poorly understood implications on the health and safety of players. EVIDENCE ACQUISITION: A literature search of the MEDLINE and PubMed databases was conducted. Keywords included athletic shoewear, cleated shoe, football shoes, and shoewear, and search parameters were between the years 2000 and 2016. STUDY DESIGN: Clinical review. LEVEL OF EVIDENCE: Level 5. RESULTS: The athletic shoe is an important piece of protective sports equipment. There are several important structural considerations of shoe design, including biomechanical compliance, cleat and turf interaction, and shoe sizing/fit, that affect the way an athlete engages with the playing surface and carry important potential implications regarding player safety if not understood and addressed. CONCLUSION: Athletic footwear should be considered an integral piece of protective equipment rather than simply an extension of the uniform apparel. More research is needed to define optimal shoe sizing, the effect that design has on mechanical load, and how cleat properties, including pattern and structure, interact with the variety of playing surfaces.

Implications of Functional Capacity Loss and Fatality for Vehicle Safety Prioritization.

OBJECTIVE: We investigate the use of the Functional Capacity Index (FCI) as a tool for establishing vehicle safety priorities by comparing the life year burden of injuries to the burden of fatality in frontal and side automotive crashes. We demonstrate FCI's utility by investigating in detail the resulting disabling injuries and their life year costs. METHODS: We selected occupants in the 2000-2013 NASS-CDS database involved in frontal and side crashes, merged their injuries with FCI, and then used the merged data to estimate each occupant's overall functional loss. Lifetime functional loss was assessed by combining this measure of impairment with the occupants' expected future life spans, estimated from the Social Security Administration's Actuarial Life Table. RESULTS: Frontal crashes produce a large number of disabling injuries, particularly to the lower extremities. In our population, these crashes are estimated to account for approximately 400,000 life years lost to disability in comparison with 500,000 life years lost to fatality. Victims of side crashes experienced a higher rate of fatality but a significantly lower rate of disabling injury (0.3 vs. 1.0%), resulting in approximately 370,000 life years lost to fatality versus 50,000 life years lost to disability. CONCLUSIONS: The burden of disabling injuries to car crash survivors should be considered when setting vehicle safety design priorities. In frontal crashes this burden in life years is similar to the burden attributable to fatality.

BioTab--a new method for analyzing and documenting injury causation in motor-vehicle crashes.

OBJECTIVE: To describe a new method for analyzing and documenting the causes of injuries in motor vehicle crashes that has been implemented since 2005 in cases investigated by the Crash Injury Research Engineering Network (CIREN). METHODS: The new method, called BioTab, documents injury causation using evidence from in-depth crash investigations. BioTab focuses on developing injury causation scenarios (ICSs) that document all factors considered essential for an injury to have occurred as well as factors that contributed to the likelihood and/or severity of an injury. The elements of an injury causation scenario are (1) the source of the energy that caused the injury, (2) involved physical components (IPCs) contacted by the occupant that are considered necessary for the injury to have occurred, (3) the body region or regions contacted by each IPC, (4) the internal paths between body regions contacted by IPCs and the injured body region, (5) critical intrusions of vehicle components, and (6) factors that contributed to the likelihood and/or the severity of injury. RESULTS: Advantages of the BioTab method are that it attempts to identify all factors that cause or contribute to clinically significant injuries, allows for coding of scenarios where one injury causes another injury, associates injuries with a source of energy and allows injuries to be associated with sources of energy other than the crash, such as air bag deployment energy, allows for documenting scenarios where an injury was caused by two different body regions contacting two different IPCs, identifies and documents the evidence that supports ICSs and IPCs, assigns confidence levels to ICSs and IPCs based on available evidence, and documents body region and organ/component-level "injury mechanisms" and distinguishes these mechanisms from ICSs. CONCLUSION: The BioTab method provides for methodical and thorough evidenced-based analysis and documentation of injury causation in motor vehicle crashes.

Thoracic response to dynamic, non-impact loading from a hub, distributed belt, diagonal belt, and double diagonal belts.

This paper presents thoracic response corridors developed using fifteen post-mortem human subjects (PMHS) subjected to single and double diagonal belt, distributed, and hub loading on the anterior thorax. We believe this is the first study to quantify the force-deflection response of the same thorax to different loading conditions using dynamic, non-impact, restraint-like loading. Subjects were positioned supine on a table and a hydraulic master-slave cylinder arrangement was used with a high-speed materials testing machine to provide controlled chest deflection at a rate similar to that experienced by restrained PMHS in a 48-km/h sled test. All loading conditions were tested at a nominally non-injurious level initially. When the battery of non-injurious tests was completed, a single loading condition was used for a final, injurious test (nominal 40% chest deflection). To minimize the influence of repeated testing, all subjects were preconditioned prior to each loading condition using 10 cycles of a 1-Hz sine wave, and the order in which the loading conditions were tested was varied across subjects. Thoracic response was characterized using the deflection at the midline of the sternum and a load cell mounted between the subject and the loading table. Responses were defined by cross-plotting the mid-sternal deflection (normalized to 50(th) male) and the posterior force (scaled to a 45-year-old, 50(th) male based on size and modulus) and then forming a +/-1-standard-deviation corridor that considered the variance in both force and deflection. Corridors were developed to a deflection level of 20% of the 50(th) percentile male's external chest depth. The distributed loading condition generated the stiffest response (3.33 kN at 4.6 cm), followed by the double diagonal belt condition (3.18 kN at 4.6 cm), the single diagonal belt (2.28 kN at 4.6 cm) and the hub (1.14 kN at 4.6 cm).