Microscale measurements of protein complexes from single cells.

Proteins execute numerous cell functions in concert with one another in protein-protein interactions (PPI). While essential in each cell, such interactions are not identical from cell to cell. Instead, PPI heterogeneity contributes to cellular phenotypic heterogeneity in health and diseases such as cancer. Understanding cellular phenotypic heterogeneity thus requires measurements of properties of PPIs such as abundance, stoichiometry, and kinetics at the single-cell level. Here, we review recent, exciting progress in single-cell PPI measurements. Novel technology in this area is enabled by microscale and microfluidic approaches that control analyte concentration in timescales needed to outpace PPI disassembly kinetics. We describe microscale innovations, needed technical capabilities, and methods poised to be adapted for single-cell analysis in the near future.

Validity and reliability of My Jump 2® app to measure the vertical jump on elite women beach volleyball players.

Purpose: The aim of this study was to assess the reliability and validity of the My Jump 2® app in measuring jump height, flight time, and peak power among elite women beach volleyball players on sand surfaces. Methods: Eleven elite female beach volleyball players (aged 23.6 ± 6.2 years; weight 66.3 ± 5.8 kg; height 174.4 ± 5.8 cm; with 8.4 ± 4.8 years of professional experience) participated in this study. Each player performed six countermovement jumps in a wooden box filled with sand on a force platform while simultaneously recording a video for subsequent analysis using the My Jump 2® app. Results: We found excellent agreement for flight time, jump height and peak power between observers (ICC = 0.92, 0.91 and 0.97, respectively). No significant differences between force platform and My Jump 2® app were detected in the values obtained for the three variables (P > 0.05). For the force platform and the My Jump 2® app, we found a good agreement measuring jump height and flight time (ICC = 0.85 and 0.85, respectively). However, we only found a moderate agreement for peak power (ICC = 0.64). The difference in jump height showed a limit of agreement between -4.10 and 4.74 cm in Bland-Altman, indicating a high level of agreement between the two measurement tools. Conclusion: Based on our findings, the My Jump 2® app reveals a valid tool for measuring jump height and flight time of CMJ on sand surfaces. However, more caution is needed when measuring peak power.

Examining the Effects of Altitude on Workload Demands in Professional Basketball Players during the Preseason Phase.

Basketball involves frequent high-intensity movements requiring optimal aerobic power. Altitude training can enhance physiological adaptations, but research examining its effects in basketball is limited. This study aimed to characterize the internal/external workload of professional basketball players during preseason and evaluate the effects of altitude and playing position. Twelve top-tier professional male basketball players (Liga Endesa, ACB; guards: n = 3, forwards: n = 5, and centers: n = 4) participated in a crossover study design composed of two training camps with nine sessions over 6 days under two different conditions: high altitude (2320 m) and sea level (10 m). Internal loads (heart rate, %HRMAX) and external loads (total distances covered across speed thresholds, accelerations/decelerations, impacts, and jumps) were quantified via wearable tracking and heart rate telemetry. Repeated-measures MANOVA tested the altitude x playing position effects. Altitude increased the total distance (+10%), lower-speed running distances (+10-39%), accelerations/decelerations (+25-30%), average heart rate (+6%), time in higher-intensity HR zones (+23-63%), and jumps (+13%) across all positions (p < 0.05). Positional differences existed, with guards accruing more high-speed running and centers exhibiting greater cardiovascular demands (p < 0.05). In conclusion, a 6-day altitude block effectively overloads training, providing a stimulus to enhance fitness capacities when structured appropriately. Monitoring workloads and individualizing training by playing position are important when implementing altitude training, given the varied responses.

Need a break? The locomotor and tackle pacing profile and loads of women's rugby league match-play following various between-match turnaround durations.

OBJECTIVES: The study investigated the locomotor and tackle pacing profile and loads of female rugby league players following various between-match turnaround durations. Specifically, the study examined the (1) pacing of locomotor and tackle loads across the time-course of a match and; (2) whole-match and peak locomotor and tackle loads of match-play. METHODS: Microtechnology data were collected from elite female rugby league players (n = 172) representing all National Rugby League Women's teams (n = 6 teams) across two seasons. Players were categorised into backs, adjustables, forwards or interchange players. Data was calculated for the whole-match (m), per minute (m.min-1) and peak (running: m.min-1; acceleration: m.s-2) locomotor and tackle loads (number and efficiency (%)) of match-play. The pacing as well as the locomotor and tackle loads of match-play were examined following short (≤6 days), normal (7 days) or long (≥8 days) turnarounds. RESULTS: The pacing profile of playing positions varied across short, normal and long match turnarounds. Trivial to moderate differences existed in the whole-match, per minute and peak locomotor loads across match turnaround durations (effect size ≤ 1.2). CONCLUSIONS: Following various between-match turnaround durations (i.e., short, normal and long match turnarounds), there were variations in the locomotor and tackle pacing profile and loads whereby, the pacing profile of positional groups was more affected than the load profile. The findings can be used in applied settings to guide the recovery strategies and training plans of female rugby league players to optimise performance and wellbeing across various match turnaround durations.

The validity of raw custom-processed global navigation satellite systems data during straight-line sprinting across multiple days.

OBJECTIVES: (1) Determine the validity of instantaneous speed and acceleration and (2) the variation in validity over time (multiple sessions) for global navigation satellite systems (GNSS) devices. DESIGN: Repeated measures. METHODS: 10-Hz GNSS devices from Statsports (n = 2, Apex Pro) and Catapult (n = 2, Vector S7) were examined, whilst a speed laser manufactured by MuscleLab (n = 1, LaserSpeed) was the criterion measure, sampling at 2.56 kHz, with data exported at 1000 Hz. Ten participants completed 40 m sprinting and changes of pace on three separate days. Root mean square error (RMSE) was used to assess the magnitude and direction of the difference between GNSS and criterion measures (instantaneous speed, instantaneous acceleration). Linear mixed models were built to assess the difference in validity across days. RESULTS: RMSE ranged from 0.14 to 0.21 m·s-1 and 0.22 to 0.47 m·s-2 for speed and acceleration, respectively showing strong agreement. There were small variations in the agreement to criterion between days for both devices for speed (Catapult RMSE = 0.12 to 21 m·s-1; Statsports RMSE = 0.14 to 0.17 m·s-1) and for acceleration (Catapult RMSE = 0.26 to 0.47 m·s-2; Statsports RMSE = 0.22 to 0.43 m·s-2) across all movements. There was a negative linear relationship between speed and acceleration error as speed increased. CONCLUSIONS: Wearable microtechnology devices from Catapult (Vector S7) and Statsports (Apex Pro) have suitable validity when measuring instantaneous speed and acceleration across multiple days. There may be small variations during different sessions and over the speed spectrum.

Silicon microfabrication technologies for biology integrated advance devices and interfaces.

Miniaturization is the trend to manufacture ever smaller devices and this process requires knowledge, experience, understanding of materials, manufacturing techniques and scaling laws. The fabrication techniques used in semiconductor industry deliver an exceptionally high yield of devices and provide a well-established platform. Today, these miniaturized devices are manufactured with high reproducibility, design flexibility, scalability and multiplexed features to be used in several applications including micro-, nano-fluidics, implantable chips, diagnostics/biosensors and neural probes. We here provide a review on the microfabricated devices used for biology driven science. We will describe the ubiquity of the use of micro-nanofabrication techniques in biology and biotechnology through the fabrication of high-aspect-ratio devices for cell sensing applications, intracellular devices, probes developed for neuroscience-neurotechnology and biosensing of the certain biomarkers. Recently, the research on micro and nanodevices for biology has been progressing rapidly. While the understanding of the unknown biological fields -such as human brain- has been requiring more research with advanced materials and devices, the development protocols of desired devices has been advancing in parallel, which finally meets with some of the requirements of biological sciences. This is a very exciting field and we aim to highlight the impact of micro-nanotechnologies that can shed light on complex biological questions and needs.

Fabrication and Investigation of Graphite-Flake-Composite-Based Non-Invasive Flex Multi-Functional Force, Acceleration, and Thermal Sensor.

This work examines the physics of a non-invasive multi-functional elastic thin-film graphite flake-isoprene sulfone composite sensor. The strain design and electrical characterization of the stretching force, acceleration, and temperature were performed. The rub-in technique was used to fabricate graphite flakes and isoprene sulfone into sensors, which were then analyzed for their morphology using methods such as SEM, AFM, X-ray diffraction, and Fourier transform infrared spectroscopy to examine the device's surface and structure. Sensor impedance was measured from DC to 200 kHz at up to 20 gf, 20 m/s2, and 26-60 °C. Sensor resistance and impedance to stretching force and acceleration at DC and 200 Hz rose 2.4- and 2.6-fold and 2.01- and 2.06-fold, respectively. Temperature-measuring devices demonstrated 2.65- and 2.8-fold decreases in resistance and impedance at DC and 200 kHz, respectively. First, altering the graphite flake composite particle spacing may modify electronic parameters in the suggested multi-functional sensors under stress and acceleration. Second, the temperature impacts particle and isoprene sulfone properties. Due to their fabrication using an inexpensive deposition technique, these devices are environmentally friendly, are simple to build, and may be used in university research in international poverty-line nations. In scientific laboratories, such devices can be used to teach students how various materials respond to varying environmental circumstances. They may also monitor individuals undergoing physiotherapy and vibrating surfaces in a controlled setting to prevent public health risks.

Recent Progress in Micro- and Nanotechnology-Enabled Sensors for Biomedical and Environmental Challenges.

Micro- and nanotechnology-enabled sensors have made remarkable advancements in the fields of biomedicine and the environment, enabling the sensitive and selective detection and quantification of diverse analytes. In biomedicine, these sensors have facilitated disease diagnosis, drug discovery, and point-of-care devices. In environmental monitoring, they have played a crucial role in assessing air, water, and soil quality, as well as ensured food safety. Despite notable progress, numerous challenges persist. This review article addresses recent developments in micro- and nanotechnology-enabled sensors for biomedical and environmental challenges, focusing on enhancing basic sensing techniques through micro/nanotechnology. Additionally, it explores the applications of these sensors in addressing current challenges in both biomedical and environmental domains. The article concludes by emphasizing the need for further research to expand the detection capabilities of sensors/devices, enhance sensitivity and selectivity, integrate wireless communication and energy-harvesting technologies, and optimize sample preparation, material selection, and automated components for sensor design, fabrication, and characterization.

External and internal maximal intensity periods of elite youth male soccer matches.

Understanding the maximal intensity periods (MIP) of soccer matches can optimise training prescription. The aim was to establish differences between positions and other contextual factors (match location, match outcome, playing formation and score line) for both external and internal MIP variables and to investigate the differences in the match start time between MIP variables. Maximal moving averages (1 to 10 min) for average speed, high-speed running (5.5-7 m·s-1), sprinting (>7 m·s-1; all m·min-1), average acceleration/deceleration (m·s-2) and heart rate (bpm, % maximal) were calculated from 24 professional youth players across 31 matches. Linear mixed models determined differences in MIP variables between positions, contextual factors and in the match start time of MIPs. Trivial to large positional differences existed in maximal external intensities while central defenders presented the lowest heart rate. It was unclear whether maximal intensities were influenced by contextual factors. MIPs for average speed, acceleration/deceleration and heart rate tend to occur concurrently (ES = trivial) within the first 30 min, while high-speed running and sprinting are likely to occur concurrently (ES = trivial) throughout a whole match. Practitioners could target maximising average speed and average acceleration/deceleration in technical-tactical based training to maximise heart rate responses.

How fast is fast? Defining velocity zones in women's rugby league.

OBJECTIVES: The study aimed to: 1) apply a data-mining approach to identify velocity zone thresholds for female rugby league players and 2) apply these velocity zones to examine the locomotor demands of match-play. METHODS: Microtechnology data were collected from elite female rugby league players representing all National Rugby League Women's teams (n = 85 players; n = 224 files) over one season. Spectral clustering with a beta smoothing cut-off of 0.1 was applied to each player's instantaneous match-play velocity data for the identification of four zones. To account for outliers within repeated data-points, the velocity zones for each player were calculated as the median. The overarching velocity zones were determined through an incremental search to minimise the root mean square error. RESULTS: Through a data-mining approach, four velocity zones were determined. Rounded to the nearest 0.5 km.h-1 the velocity values across each zone were classified as low (<11.49 km.h-1), moderate (11.50 to 17.49 km.h-1), high (17.50 to 20.99 km.h-1) and very-high (>21.00 km.h-1). Practical application of the zones demonstrated positional group differences in the absolute (effect size (ES): 0.03 to 1.77) and relative (ES: 0.04 to 1.60) locomotor demands of match-play. The back positional group covered greater absolute and relative distances at a very-high velocity than all other positions. CONCLUSIONS: This work informs the velocity zones that could be applied consistently to women's rugby league data within practical (i.e., in the training and monitoring of players) and academic (i.e., as a model for future research to analyse locomotor demands) settings.

Locomotor and technical characteristics of female soccer players training: exploration of differences between competition standards.

OBJECTIVES: To (i) quantify the differences in locomotor and technical characteristics between different drill categories in female soccer and (ii) explore the training drill distributions between different standards of competition. METHODS: Technical (ball touches, ball releases, high-speed ball releases) and locomotor data (total distance, high-speed running distance [>5.29 m∙s-1]) were collected using foot-mounted inertial measurement units from 458 female soccer players from three Women's Super League (WSL; n = 76 players), eight Women's Championship (WC; n = 217) and eight WSL Academy (WSLA; n = 165) teams over a 28-week period. Data were analysed using general linear mixed effects. RESULTS: Across all standards, the largest proportion of time was spent in technical (TEC) (WSL = 38%, WC = 28%, WSLA = 29%) and small-sided extensive games (SSGe) (WSL = 20%, WC = 31%, WSLA = 30%) drills. WSL completed more TEC and tactical (TAC) training whilst WC and WSLA players completed more SSGe and possession (POS) drills. Technical drills elicited the highest number of touches, releases and the highest total distance and high-speed activity. Position-specific drills elicited the lowest number of touches and releases and the lowest total distance. When the technical and locomotor demand of each drill were made relative to time, there were limited differences between drills, suggesting drill duration was the main moderating factor. CONCLUSION: Findings provide novel understanding of the technical and locomotor demands of different drill categories in female soccer. These results can be used by coaches and practitioners to inform training session design.

Noninvasive prenatal diagnosis targeting fetal nucleated red blood cells.

Noninvasive prenatal diagnosis (NIPD) aims to detect fetal-related genetic disorders before birth by detecting markers in the peripheral blood of pregnant women, holding the potential in reducing the risk of fetal birth defects. Fetal-nucleated red blood cells (fNRBCs) can be used as biomarkers for NIPD, given their remarkable nature of carrying the entire genetic information of the fetus. Here, we review recent advances in NIPD technologies based on the isolation and analysis of fNRBCs. Conventional cell separation methods rely primarily on physical properties and surface antigens of fNRBCs, such as density gradient centrifugation, fluorescence-activated cell sorting, and magnetic-activated cell sorting. Due to the limitations of sensitivity and purity in Conventional methods, separation techniques based on micro-/nanomaterials have been developed as novel methods for isolating and enriching fNRBCs. We also discuss emerging methods based on microfluidic chips and nanostructured substrates for static and dynamic isolation of fNRBCs. Additionally, we introduce the identification techniques of fNRBCs and address the potential clinical diagnostic values of fNRBCs. Finally, we highlight the challenges and the future directions of fNRBCs as treatment guidelines in NIPD.

Changing gears: data-driven velocity zones to support monitoring and research in men's rugby league.

OBJECTIVES: The study aimed to (1) apply a data-mining approach to league-wide microtechnology data to identify absolute velocity zone thresholds and (2) apply the respective velocity zones to microtechnology data to examine the locomotor demands of elite match-play. METHODS: League-wide microtechnology data were collected from elite male rugby league players representing all National Rugby League (NRL) teams (n = 16 teams, one excluded due to a different microtechnology device; n = 4836 files) over one season. To identify four velocity zones, spectral clustering with a beta smoothing cut-off of 0.1 was applied to each players' instantaneous match-play velocity data. Velocity zones for each player were calculated as the median while the overarching velocity zones were determined through an incremental search to minimise root mean square error. RESULTS: The velocity zones identified through spectral clustering were 0-13.99 km · h-1 (i.e., low velocity), 14.00-20.99 km · h-1 (i.e., moderate velocity), 21.00-24.49 km · h-1 (i.e., high velocity) and >24.50 km · h-1 (i.e., very-high velocity). CONCLUSIONS: The application of spectral clustering (i.e., a data-mining method) to league-wide rugby league microtechnology data yielded insights into the distribution of velocity data, thereby informing the cut-off values which best place similar data points into the same velocity zones. As the identified zones are representative of the intensities of locomotion achieved by elite male rugby league players, it is suggested that when absolute zones are used, the consistent application of the identified zones would facilitate standardisation, longitudinal athlete monitoring as well as comparisons between teams, leagues and published literature.

Concurrent Validity and Reliability of Devices to Measure Jump Height in Men's Handball Players.

Although there is a wide range of validated devices to measure vertical jump height, the degree of interchangeability among them is currently unknown. AIMS: The purpose of this study was to examine the concurrent validity and reliability of multiple devices to measure jump height in men's handball players. METHODS: Sixteen players (age = 24.0 ± 3.7 years old) performed three types of jumps (n= 144-squat jump (SJ), countermovement jump (CMJ) and Abalakov jump (ABK)) on a contact platform (CHRONOJUMP®) while simultaneously being measured with two inertial devices (WIMU® and VERT®) and recorded with a high-speed camera. Vertical jump height was analyzed according to each type of jump. RESULTS: The t-test showed statistically significant differences (p = 0.001) between the contact platform (reference standard) and the rest of the tools that tended to overestimate jump height in all jumps. SJ and CMJ proved to be the jump tests with the most stable reliability values in all devices (ICC: 0.92-0.98), except in the comparison with VERT®. CONCLUSIONS: Although all the analyzed devices proved to be valid and reliable in previous studies, they are not interchangeable. Therefore, it is suggested to always use the same type of device to evaluate vertical height jump.

Tackle Technique and Changes in Playerload™ During a Simulated Tackle: An Exploratory Study.

In collision sports, the tackle has the highest injury incidence, and is key to a successful performance. Although the contact load of players has been measured using microtechnology, this has not been related to tackle technique. The aim of this study was to explore how PlayerLoad™ changes between different levels of tackling technique during a simulated tackle. Nineteen rugby union players performed twelve tackles on a tackle contact simulator (n = 228 tackles). Each tackle was recorded with a video-camera and each player wore a Catapult OptimEyeS5. Tackles were analysed using tackler proficiency criteria and split into three categories: Low scoring(≤5 Arbitrary units (AU), medium scoring(6 and 7AU) and high scoring tackles(≥8AU). High scoring tackles recorded a higher PlayerLoad™ at tackle completion. The PlayerLoad™ trace was also less variable in the high scoring tackles. The variability in the PlayerLoad™ trace may be a consequence of players not shortening their steps before contact. This reduced their ability to control their movement during the contact and post-contact phase of the tackle and increased the variability. Using the PlayerLoad™ trace in conjunction with subjective technique assessments offers coaches and practitioners insight into the physical-technical relationship of each tackle to optimise tackle skill training and match preparation.

Assessment of Relationships Between External Load Metrics and Game Performance in Women's Lacrosse.

The purpose of this study was to analyse the relationship between external load metrics with game performance for Division I collegiate female lacrosse. Data were collected using microtechnology during 26 games with 13 athletes (attackers n = 5, midfielders n = 8). External load variables included: total distance (TD), distance rate (DR), high-intensity (HI) distance, speed, HI sprints, accelerations, decelerations, metabolic equivalent distance (MED), and sprints. For attackers, goals, points, shots, and shots on goal (SOG) had low, positive associations with TD (r = 0.32 to 0.42) and MED (r = 0.39 to 0.45). For midfielders, goals, shots, and draw controls had low, positive relationships with TD (r = 0.34 to 0.41), DR (r = 0.33 to 0.45), and decelerations (r = 0.30 to 0.35). Points and SOG had low associations with TD (r = 0.35), DR (r = 0.33), accelerations (r = 0.31), and decelerations (r = 0.32 to 0.35). Turnovers demonstrated low, positive relationships with HI distance (r = 0.31) and HI sprints (r = 0.41). Though the correlations were low, they still revealed insights into workload metrics and offensive game statistics in the sport of lacrosse. Training for attackers should focus on agility and a training base for TD. Training for midfielders should focus on HI endurance and sprinting.

Therapeutic implementation in arterial thrombosis with pulmonary administration of fucoidan microparticles containing acetylsalicylic acid.

Several antithrombotic drugs are available to treat cardiovascular diseases due to its high mortality and morbidity worldwide. Despite these, severe adverse effects that can lead to treatment withdrawal have been described, highlighting the importance of new therapies. Thus, this work describes the development of fucoidan microparticles containing acetylsalicylic acid (MP/F4M) for pulmonary delivery and in vitro, ex vivo, and in vivo evaluation. Microparticles were prepared via spray-drying and characterized in vitro (mucoadhesive properties, coagulation time, platelet aggregation, adhesion, and hemolysis) followed by ex vivo platelet aggregation, in vivo arterial thrombosis, and hemorrhagic profile. The formulation physicochemical characterization showed suitable characteristics along with delayed drug release, increased breathable particle fraction, and high washability resistance as well as antiplatelet activity and enhanced platelet adhesion in vitro. In in vivo assays, MP/F4M protected against arterial thrombosis, without changes in the hemorrhagic profile. Finally, no lung changes were observed after prolonged pulmonary administration, whereas isolated ASA led to an inflammatory response. In conclusion, pulmonary administration of fucoidan microparticles with an antiplatelet drug may be an alternative therapy to treat cardiovascular diseases, opening the field for different formulations.

Quantifying technical actions in professional soccer using foot-mounted inertial measurement units.

OBJECTIVES: This study aimed to (i) establish the concurrent validity and intra-unit reliability of a foot-mounted inertial measurement unit for monitoring soccer technical actions, (ii) quantify the within-microcycle inter-positional differences in the technical actions of professional soccer training, and (iii) determine the influence of drill category on the technical actions of professional soccer training. METHODS: Twenty-one professional soccer players' technical performance data (ball touches, releases, ball touches per minute, releases per minute), collected during training sessions throughout twenty-four weekly microcycles, were analysed using general linear modelling. RESULTS: The inertial measurement unit exhibited good concurrent validity (PA = 95.1% - 100.0%) and intra-unit reliability (PA = 95.9% - 96.9%, CV = 1.4% - 2.9%) when compared with retrospective video analyses. The most ball touches (X‾ = 218.0) and releases (X‾ = 110.8) were observed on MD - 1, with MD - 5 eliciting the highest frequency of ball touches (X‾ = 3.8) and releases (X‾ = 1.7) per minute. Central midfielders performed the most ball touches (X‾ = 221.9), releases (X‾ = 108.3), ball touches per minute (X‾ = 3.4) and releases per minute (X‾ = 1.6). Small-sided games evoked more ball touches (X‾diff = 1.5) and releases per minute (X‾diff = 0.1) than previously reported in match-play. The fewest ball touches (X‾ = 1.2) and releases per minute (X‾ = 0.5) were observed during tactical drills. CONCLUSION: The results of this study provide a novel understanding of the within-microcycle, inter-positional and drill category differences in the technical actions performed by professional players during training.

Remote Sensing System for Motor Nerve Impulse.

In this article, we present our research achievements regarding the development of a remote sensing system for motor pulse acquisition, as a first step towards a complete neuroprosthetic arm. We present the fabrication process of an implantable electrode for nerve impulse acquisition, together with an innovative wirelessly controlled system. In our study, these were combined into an implantable device for attachment to peripheral nerves. Mechanical and biocompatibility tests were performed, as well as in vivo testing on pigs using the developed system. This testing and the experimental results are presented in a comprehensive manner, demonstrating that the system is capable of accomplishing the requirements of its designed application. Most significantly, neural electrical signals were acquired and transmitted out of the body during animal experiments, which were conducted according to ethical regulations in the field.

Derivation and Differentiation of Human Pluripotent Stem Cells in Microfluidic Devices.

An integrative approach based on microfluidic design and stem cell biology enables capture of the spatial-temporal environmental evolution underpinning epigenetic remodeling and the morphogenetic process. We examine the body of literature that encompasses microfluidic applications where human induced pluripotent stem cells are derived starting from human somatic cells and where human pluripotent stem cells are differentiated into different cell types. We focus on recent studies where the intrinsic features of microfluidics have been exploited to control the reprogramming and differentiation trajectory at the microscale, including the capability of manipulating the fluid velocity field, mass transport regime, and controllable composition within micro- to nanoliter volumes in space and time. We also discuss studies of emerging microfluidic technologies and applications. Finally, we critically discuss perspectives and challenges in the field and how these could be instrumental for bringing about significant biological advances in the field of stem cell engineering.