Control-Normalized Fisher Ratio Analysis of Comprehensive Two-Dimensional Gas Chromatography Time-of-Flight Mass Spectrometry Data for Enhanced Biomarker Discovery in a Metabolomic Study of Orthopedic Knee-Ligament Injury.

An innovative form of Fisher ratio (F-ratio) analysis (FRA) is developed for use with comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GC × GC-TOFMS) data and applied to the investigation of the changes in the metabolome in human plasma for patients with injury to their anterior cruciate ligament (ACL). Specifically, FRA provides a supervised discovery of metabolites that express a statistically significant variance in a two-sample class comparison: patients and healthy controls. The standard F-ratio utilizes the between-class variance relative to the pooled within-class variance. Because standard FRA is adversely impacted by metabolites expressed with a large within-class variance in the patient class, "control-normalized FRA" has been developed to provide complementary information, by normalizing the between-class variance to the variance of the control class only. Thirty plasma samples from patients who recently suffered from an ACL injury, along with matched controls, were subjected to GC × GC-TOFMS analysis. Following both standard and control-normalized FRA, the concentration ratio for the top 30 "hits" in each comparison was obtained and then t-tested for statistical significance. Twenty four out of 30 metabolites plus the therapeutic agent, naproxen (24/30), passed the t-test for the control-normalized FRA, which included 8/24 unique to control-normalized FRA and 16/24 in common with the standard FRA. Likewise, standard FRA provided 21/30 metabolites passing the t-test, with 5/21 undiscovered by control-normalized FRA. The complementary information obtained by both F-ratio analyses demonstrates the general utility of the new approach for a variety of applications.

Progress and Future Directions of the NCAA-DoD Concussion Assessment, Research, and Education (CARE) Consortium and Mind Matters Challenge at the US Service Academies.

Despite the significant impact that concussion has on military service members, significant gaps remain in our understanding of the optimal diagnostic, management, and return to activity/duty criteria to mitigate the consequences of concussion. In response to these significant knowledge gaps, the US Department of Defense (DoD) and the National Collegiate Athletic Association (NCAA) partnered to form the NCAA-DoD Grand Alliance in 2014. The NCAA-DoD CARE Consortium was established with the aim of creating a national multisite research network to study the clinical and neurobiological natural history of concussion in NCAA athletes and military Service Academy cadets and midshipmen. In addition to the data collected for the larger CARE Consortium effort, the service academies have pursued military-specific lines of research relevant to operational and medical readiness associated with concussion. The purpose of this article is to describe the structure of the NCAA-DoD Grand Alliance efforts at the service academies, as well as discuss military-specific research objectives and provide an overview of progress to date. A secondary objective is to discuss the challenges associated with conducting large-scale studies in the Service Academy environment and highlight future directions for concussion research endeavors across the CARE Service Academy sites.

Trunk and Lower Extremity Movement Patterns, Stress Fracture Risk Factors, and Biomarkers of Bone Turnover in Military Trainees.

CONTEXT: Military service members commonly sustain lower extremity stress fractures (SFx). How SFx risk factors influence bone metabolism is unknown. Understanding how SFx risk factors influence bone metabolism may help to optimize risk-mitigation strategies. OBJECTIVE: To determine how SFx risk factors influence bone metabolism. DESIGN: Cross-sectional study. SETTING: Military service academy. PATIENTS OR OTHER PARTICIPANTS: Forty-five men (agepre = 18.56 ± 1.39 years, heightpre = 176.95 ± 7.29 cm, masspre = 77.20 ± 9.40 kg; body mass indexpre = 24.68 ± 2.87) who completed Cadet Basic Training (CBT). Individuals with neurologic or metabolic disorders were excluded. INTERVENTION(S): We assessed SFx risk factors (independent variables) with (1) the Landing Error Scoring System (LESS), (2) self-reported injury and physical activity questionnaires, and (3) physical fitness tests. We assessed bone biomarkers (dependent variables; procollagen type I amino-terminal propeptide [PINP] and cross-linked collagen telopeptide [CTx-1]) via serum. MAIN OUTCOME MEASURE(S): A markerless motion-capture system was used to analyze trunk and lower extremity biomechanics via the LESS. Serum samples were collected post-CBT; enzyme-linked immunosorbent assays determined PINP and CTx-1 concentrations, and PINP : CTx-1 ratios were calculated. Linear regression models demonstrated associations between SFx risk factors and PINP and CTx-1 concentrations and PINP : CTx-1 ratio. Biomarker concentration mean differences with 95% confidence intervals were calculated. Significance was set a priori using α ≤ .10 for simple and α ≤ .05 for multiple regression analyses. RESULTS: The multiple regression models incorporating LESS and SFx risk factor data predicted the PINP concentration (R2 = 0.47, P = .02) and PINP : CTx-1 ratio (R2 = 0.66, P = .01). The PINP concentration was increased by foot internal rotation, trunk flexion, CBT injury, sit-up score, and pre- to post-CBT mass changes. The CTx-1 concentration was increased by heel-to-toe landing and post-CBT mass. The PINP : CTx-1 ratio was increased by foot internal rotation, lower extremity sagittal-plane displacement (inversely), CBT injury, sit-up score, and pre- to post-CBT mass changes. CONCLUSIONS: Stress fracture risk factors accounted for 66% of the PINP : CTx-1 ratio variability, a potential surrogate for bone health. Our findings provide insight into how SFx risk factors influence bone health. This information can help guide SFx risk-mitigation strategies.

Muscle inflammation susceptibility: a prognostic index of recovery potential after hip arthroplasty?

While elective total hip arthroplasty (THA) for end-stage osteoarthritis (OA) improves pain, mobility function, and quality of life in most cases, a large proportion of patients suffer persistent muscle atrophy, pain, and mobility impairment. Extensive skeletal muscle damage is unavoidable in these surgical procedures, and it stands to reason that poor recovery and long-term mobility impairment among some individuals after THA is linked to failed muscle regeneration and regrowth following surgery and that local muscle inflammation susceptibility (MuIS) is a major contributing factor. Here we present results of two integrated studies. In study 1, we compared muscle inflammation and protein metabolism signaling in elective THA (n=15) vs. hip fracture/trauma (HFX; n=11) vs. nonsurgical controls (CON; n=19). In study 2, we compared two subgroups of THA patients dichotomized into MuIS⁺ (n=7) or MuIS⁻ (n=7) based on muscle expression of TNF-like weak inducer of apoptosis (TWEAK) receptor (Fn14). As expected, HFX demonstrated overt systemic and local muscle inflammation and hypermetabolism. By contrast, no systemic inflammation was detected in elective THA patients; however, local muscle inflammation in the perioperative limb was profound in MuIS⁺ and was accompanied by suppressed muscle protein synthesis compared with MuIS⁻. Muscle from the contralateral limb of MuIS⁺ was unaffected, providing evidence of a true inflammation susceptibility localized to the muscle surrounding the hip with end-stage OA. We suggest MuIS status assessed at the time of surgery may be a useful prognostic index for muscle recovery potential and could therefore provide the basis for a personalized approach to postsurgery rehabilitation.