Citations

Glioma cell migration correlates with Pyk2 activity, but the intrinsic mechanism that regulates the activity of Pyk2 is not fully understood. Previous studies have supported a role for the N-terminal FERM domain in the regulation of Pyk2 activity as mutations in the FERM domain inhibit Pyk2 phosphorylation. To search for novel protein-protein interactions mediated by the Pyk2 FERM domain, we utilized a yeast two-hybrid genetic selection to identify the mammalian Ste20 homolog MAP4K4 as a binding partner for the Pyk2 FERM domain. MAP4K4 coimmunoprecipitated with Pyk2 and was a substrate for Pyk2 but did not coimmunoprecipitate with the closely related focal adhesion kinase FAK. Knockdown of MAP4K4 expression inhibited glioma cell migration and effectively blocked Pyk2 stimulation of glioma cell. Increased expression of MAP4K4 stimulated glioma cell migration; however, this stimulation was blocked by knockdown of Pyk2 expression. These data support that the interaction of MAP4K4 and Pyk2 is integrated with glioma cell migration and suggest that inhibition of this interaction may represent a potential therapeutic strategy to limit glioblastoma tumor dispersion.

Background— Cardiac development is a complex process resulting in an integrated, multilineage tissue with developmental corruption in early embryogenesis leading to congenital heart disease. Interrogation of individual genes has provided the backbone for cardiac developmental biology, yet a comprehensive transcriptome derived from natural cardiogenesis is required to gauge innate developmental milestones. Methods and Results— Stage-specific cardiac structures were dissected from 8 distinctive mouse embryonic time points to produce genome-wide expressome analysis across cardiogenesis. With reference to this native cardiogenic expression roadmap, divergent induced pluripotent stem cell–derived cardiac expression profiles were mapped from procardiogenic 3-factor (SOX2, OCT4, KLF4) and less-cardiogenic 4-factor (plus c-MYC) reprogrammed cells. Expression of cardiac-related genes from 3-factor–induced pluripotent stem cell differentiated in vitro at days 5 and 11 and recapitulated expression profiles of natural embryos at days E7.5-E8.5 and E14.5-E18.5, respectively. By contrast, 4-factor–induced pluripotent stem cells demonstrated incomplete cardiogenic gene expression profiles beginning at day 5 of differentiation. Differential gene expression within the pluripotent state revealed 23 distinguishing candidate genes among pluripotent cell lines with divergent cardiogenic potentials. A confirmed panel of 12 genes, differentially expressed between high and low cardiogenic lines, was transformed into a predictive score sufficient to discriminate individual induced pluripotent stem cell lines according to relative cardiogenic potential. Conclusions— Transcriptome analysis attuned to natural embryonic cardiogenesis provides a robust platform to probe coordinated cardiac specification and maturation from bioengineered stem cell–based model systems. A panel of developmental-related genes allowed differential prognosis of cardiogenic competency, thus prioritizing cell lines according to natural blueprint to streamline functional applications.

ABSTRACTIntroduction: Repetitive discharges may be recorded during nerve conduction studies (NCS) or during needle electromyography in a muscle at rest. Repetitive discharges that occur during voluntary activation and are time‐locked to voluntary motor unit potentials (MUP) have not been described. Methods: Retrospective review of motor unit potential induced repetitive discharges (MIRDs) identified in the EMG laboratory. Characteristics of each MIRD, patient demographics, other EMG findings in the same muscle, and electrophysiological diagnosis were analyzed. Results: MIRDs were observed in 15 patients. The morphology and number of spikes and duration of MIRDs varied. The discharges fired at rates of 50–200 Hz. All but 2 patients had EMG findings of a chronic neurogenic disorder. Conclusions: MIRDs are rare iterative discharges time‐locked to a voluntary MUP. The pathophysiology of MIRDs is unclear, but their presence may indicate a chronic neurogenic process. Muscle Nerve 48: 977–979, 2013

Objective Although the majority of high altitude studies have used doubly labeled water to assess energy expenditure (EE), minimal research has examined variation in intensity levels while participating in such an expedition. The purpose of this study was to use ambulatory monitoring technology to assess EE and time spent in moderate (3.0–6.0 METS), vigorous (6.0–9.0 METS), and very vigorous (>9.0 METS) intensities during an expedition to summit a 6170-m peak in Northern Pakistan. Methods Twelve men and 2 women were studied. Their EE was determined from galvanic skin response, heat flux, and biaxial accelerometry using a multisensor armband. Lake Louise Scores for altitude illness assessment were obtained each morning. Results Average daily total EE and number of steps taken was 4173 ± 848 kcal/d (range, 3332 to 4838 kcal/d) and 26,232 ± 7210 steps/d (range, 11,788 to 34,979 steps/d), respectively. On average, subjects spent approximately 7.7 h/d performing physical activity (>3.0 METS). Of this, subjects spent 399 ± 107, 61 ± 77, and 3 ± 10 min/d at moderate, vigorous, and very vigorous intensity levels, respectively, corresponding to 88%, 11%, and 1% of total physical activity time. During summit day, subjects climbed at an intensity equivalent to approximately 42% of their high altitude predicted maximal oxygen consumption at 5100 m). Conclusions During a high altitude expedition, subjects predominantly exercise at a moderate intensity, with limited time spent at higher intensity levels. Use of accelerometer-based technology may provide more cost-effective and logistically easier means of assessing EE in extreme environments.

SUMMARYFibrosis of the subsynovial connective tissue (SSCT) in the carpal tunnel is the most common histological finding in carpal tunnel syndrome (CTS). Fibrosis may result from damaged SSCT. Previous studies found that with low‐velocity (2 mm/s), tendon excursions can irreversibly damage the SSCT. We investigated the effect of tendon excursion velocity in the generation of SSCT damage. Nine human cadaver wrists were used. Three repeated cycles of ramp‐stretch testing were performed simulating 40%, 60%, 90%, and 120% of the middle finger flexor tendon superficialis physiological excursion with an excursion velocity of 60 mm/s. Energy and force were calculated and normalized by values obtained in the first cycle for each excursion level. Data were compared with low‐velocity excursion data. For high‐velocity excursions, a significant drop in the excursion energy ratio was first observed at an excursion level of 60% physiological excursion (p < 0.024) and that for low‐velocity excursions was first observed at 90% physiological excursion (p < 0.038). Furthermore, the energy ratio was lower at 60% for high velocities (p ≤ 0.039). Increasing velocity lowers the SSCT damage threshold. This finding may be relevant for understanding the pathogenesis of SSCT fibrosis, such as that accompanying CTS, and a relationship with occupational factors. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:123–128, 2014.

AbstractMutations in superoxide dismutase 1 (SOD1) associated with familial amyotrophic lateral sclerosis induce misfolding and aggregation of the protein with the inherent propensity of mutant SOD1 to aggregate generally correlating, with a few exceptions, to the duration of illness in patients with the same mutation. One notable exception was the D101N variant, which has been described as wild‐type‐like. The D101N mutation is associated with rapidly progressing motor neuron degeneration but shows a low propensity to aggregate. By assaying the kinetics of aggregation in a well‐characterized cultured cell model, we show that the D101N mutant is slower to initiate aggregation than the D101G mutant. In this cell system of protein over‐expression, both mutants were equally less able to acquire Zn than WT SOD1. In addition, both of these mutants were equivalently less able to fold into the trypsin‐resistant conformation that characterizes WT SOD1. A second major difference between the two mutants was that the D101N variant more efficiently formed a normal intramolecular disulfide bond. Overall, our findings demonstrate that the D101N and D101G variants exhibit clearly distinctive features, including a different rate of aggregation, and yet both are associated with rapidly progressing disease. image We sought to better characterize the biochemical features of two SOD1 mutants associated with rapidly progressing disease, the D101G and wild‐type like D101N mutants. We observed using our cell model that that although similarities were observed when comparing the ability to bind metals and resist trypsin digestion, these mutants differed in their ability to initiate aggregation and to form the normal intramolecular disulfide bond. We conclude that these mutants exhibit distinct properties despite producing similar disease phenotypes in patients.

Background: We used simulation modeling to relate glucose meter performance criteria to insulin dosing errors for patients on a moderate glycemic control protocol (glucose target, 110–150 mg/dL) and empirically validated assumptions from simulation models using observed glucose meter and laboratory glucose values obtained nearly simultaneously. Subjects and Methods: The 25,948 glucose values from 1,513 patients on a moderate glycemic control protocol were used to represent the expected distribution of glucose values in this patient population. Simulation models were used to relate glucose meter analytical performance to insulin dosing errors assuming 10%, 15%, or 20% total allowable error (TEa). In addition, 4,017 paired glucose meter and serum laboratory glucose measurements drawn within 5 min of each other were used to generate an empiric dataset to validate simulation model assumptions relating glucose meter performance to insulin dosing errors. Results: Large (three or more category) insulin dosing errors are predicted to occur only under the 20% TEa condition. Two category insulin dosing errors were common (6–20% of all insulin dosing decisions) when 20% TEa was assumed, but frequency decreased to only 0.2% of dosing decisions when 10% TEa was modeled. When insulin dosing error rates were measured empirically by comparing paired glucose meter and laboratory glucose values, insulin dosing error rates were very similar to those predicted for the 20% TEa condition. Conclusions: Both simulation models and empiric data demonstrate that glucose meters that perform at ≥20% TEa allow large insulin dosing errors during a moderate glycemic control protocol.