Analysis of Injury Metrics From Experimental Cardiac Injuries From Behind Armor Blunt Trauma Using Live Swine Tests: A Pilot Study.

INTRODUCTION: Warfighters are issued hard body armor designed to defeat ballistic projectiles. The resulting backface deformation can injure different thoracoabdominal organs. Developed over decades ago, the behind armor blunt impact criterion of maximum 44 mm depth in clay continues to be used independent of armor type or impact location on the thoracoabdominal region covered by the armor. Because thoracoabdominal components have different energy absorption capabilities, their mode of failures and mechanical properties are different. These considerations underscore the lack of effectiveness of using the single standard to cover all thoracoabdominal components to represent the same level of injury risk. The objective of this pilot study is to conduct cardiac impact tests with a live animal model and analyze biomechanical injury candidate metrics for behind armor blunt trauma applications. MATERIALS AND METHODS: Live swine tests were conducted after obtaining approvals from the U.S. DoD. Trachea tubes. An intravenous line were introduced into the swine before administering anesthesia. Pressure transducers were inserted into lungs and aorta. An indenter simulating backface deformation profiles produced by body armor from military-relevant ballistics to human cadavers delivered impact to the heart region. The approved test protocol included 6-hour monitoring and necropsies. Indenter accelerometer signals were processed to compute the velocity and deflection, and their peak magnitudes were obtained. The deflection-time signal was normalized with respect to chest depth along the impact axis. The peak magnitude of the viscous criterion, kinetic energy, force, momentum and stiffness were obtained. RESULTS: Out of the 8 specimens, 2 were sham controls. The mean total body mass and soft tissue thickness at the impact site were 81.1 ± 4.1 kg and 3.8 ± 1.1 cm. The peak velocities ranged from 30 to 59 m/s, normalized deflections ranged from 15 to 21%, and energies ranged from 105 to 407 J. The range in momentum and stiffness were 7.0 to 13.9 kg-m/s and 22.3 to 79.9 N/m. The maximum forces and impulse data ranged from 2.9 to 11.7 kN and 1.9 to 5.8 N-s. The peak viscous criterion ranged from 2.0 to 5.3 m/s. One animal did not sustain any injuries, 2 had cardiac injuries, and others had lung and skeletal injuries. CONCLUSIONS: The present study applied blunt impact loads to the live swine cardiac region and determined potential candidate injury metrics for characterization. The sample size of 6 swine produced injuries ranging from none to pure skeletal to pure organ trauma. The viscous criterion metric associated with the response of the animal demonstrated a differing pattern than other variables with increasing velocity. These findings demonstrate that our live animal experimental design can be effectively used with testing additional samples to develop behind armor blunt injury criteria for cardiac trauma in the form of risk curves. Injury criteria obtained for cardiac trauma can be used to enhance the effectiveness of the body armor, reduce morbidity and mortality, and improve warfighter readiness in combat operations.

Investigating the Impact of Blunt Force Trauma: A Probabilistic Study of Behind Armor Blunt Trauma Risk.

Evaluating Behind Armor Blunt Trauma (BABT) is a critical step in preventing non-penetrating injuries in military personnel, which can result from the transfer of kinetic energy from projectiles impacting body armor. While the current NIJ Standard-0101.06 standard focuses on preventing excessive armor backface deformation, this standard does not account for the variability in impact location, thorax organ and tissue material properties, and injury thresholds in order to assess potential injury. To address this gap, Finite Element (FE) human body models (HBMs) have been employed to investigate variability in BABT impact conditions by recreating specific cases from survivor databases and generating injury risk curves. However, these deterministic analyses predominantly use models representing the 50th percentile male and do not investigate the uncertainty and variability inherent within the system, thus limiting the generalizability of investigating injury risk over a diverse military population. The DoD-funded I-PREDICT Future Naval Capability (FNC) introduces a probabilistic HBM, which considers uncertainty and variability in tissue material and failure properties, anthropometry, and external loading conditions. This study utilizes the I-PREDICT HBM for BABT simulations for three thoracic impact locations-liver, heart, and lower abdomen. A probabilistic analysis of tissue-level strains resulting from a BABT event is used to determine the probability of achieving a Military Combat Incapacitation Scale (MCIS) for organ-level injuries and the New Injury Severity Score (NISS) is employed for whole-body injury risk evaluations. Organ-level MCIS metrics show that impact at the heart can cause severe injuries to the heart and spleen, whereas impact to the liver can cause rib fractures and major lacerations in the liver. Impact at the lower abdomen can cause lacerations in the spleen. Simulation results indicate that, under current protection standards, the whole-body risk of injury varies between 6 and 98% based on impact location, with the impact at the heart being the most severe, followed by impact at the liver and the lower abdomen. These results suggest that the current body armor protection standards might result in severe injuries in specific locations, but no injuries in others.

Matched-pair hybrid test paradigm for behind armor blunt trauma using an experimental animal model.

Background: The current behind armor blunt trauma (BABT) injury criterion uses a single penetration limit of 44 mm in Roma Plastilina clay and is not specific to thoracoabdominal regions. However, different regions in the human body have different injury tolerances. This manuscript presents a matched-pair hybrid test paradigm with different experimental models and candidate metrics to develop regional human injury criteria. Methods: Live and cadaver swine were used as matched pair experimental models. An impactor simulating backface deformation profiles produced by body armor from military-relevant ballistics was used to deliver BABT loading to liver and lung regions in cadaver and live swine. Impact loading was characterized using peak accelerations and energy. For live swine, physiological parameters were monitored for 6 hours, animals were euthanized, and a detailed necropsy was done to identify injuries to skeletal structures, organs and soft tissues. A similar process was used to identify injuries to the cadaver swine for targeted thoracoabdominal regions. Results: Two cadavers and one live swine were subjected to BABT impacts to the liver. One cadaver and one live swine were subjected to BABT impacts to the left lung. Injuries to both regions were similar at similar energies between the cadaver and live models. Conclusions: Swine is an established animal for thoracoabdominal impact studies in automotive standards, although at lower insult levels. Similarities in BABT responses between cadaver and live swine allow for extending testing protocols to human cadavers and for the development of scaling relationships between animal and human cadavers, acting as a hybrid protocol between species and live and cadaver models. Injury tolerances and injury risk curves from live animals can be converted to human tolerances via structural scaling using these outcomes. The present experimental paradigm can be used to develop region-based BABT injury criteria, which are not currently available.

Efficacy and Safety of Microsomal Ferric Pyrophosphate Supplement for Iron Deficiency Anemia in Pregnancy.

Introduction Anemia during pregnancy is characterized by decreased hemoglobin levels. Iron deficiency poses a significant global health concern, especially in pregnant women, where increased iron demands are crucial for both maternal and fetal well-being. Method In the current study, we investigated the effectiveness and safety of 30 mg SunActiveTM Fe (Taiyo GmbH, Yokkaichi, Japan), emulsified microsomalTM ferric pyrophosphate (EMFP) tablets in treating iron deficiency anemia in 27 second-trimester singleton pregnant women. Results Our study results demonstrated that hemoglobin levels increased significantly within 30 days of treatment and continued to remain higher than baseline throughout the study. Serum ferritin levels exhibited a 6.61-fold increase, maintaining elevated levels consistently. Serum iron also increased significantly by 46.9%. Additionally, symptoms such as nausea, breathlessness, dizziness, irritability, and heartburn were notably reduced, leading to improved quality of life. Subjects reported decreased overall fatigue, indicating an enhanced quality of life. Babies born during the study showed healthy birth weights, with uncomplicated deliveries. High treatment compliance of 99.5% underscored patient commitment to the study. Furthermore, the investigational product demonstrated a favorable safety profile, with only two mild adverse events observed, unrelated to the treatment. Conclusion These findings suggest that EMFP could be a valuable therapeutic option for managing iron deficiency anemia in pregnant women, promoting better maternal and fetal outcomes. Further research with an increased sample size is warranted to delve into the underlying mechanisms behind these positive outcomes, nonetheless, our study provides a promising foundation for addressing this critical health issue.

Subject-Specific Geometry of FE Lumbar Spine Models for the Replication of Fracture Locations Using Dynamic Drop Tests.

For traumatic lumbar spine injuries, the mechanisms and influence of anthropometrical variation are not yet fully understood under dynamic loading. Our objective was to evaluate whether geometrically subject-specific explicit finite element (FE) lumbar spine models based on state-of-the-art clinical CT data combined with general material properties from the literature could replicate the experimental responses and the fracture locations via a dynamic drop tower-test setup. The experimental CT datasets from a dynamic drop tower-test setup were used to create anatomical details of four lumbar spine models (T12 to L5). The soft tissues from THUMS v4.1 were integrated by morphing. Each model was simulated with the corresponding loading and boundary conditions from the dynamic lumbar spine tests that produced differing injuries and injury locations. The simulations resulted in force, moment, and kinematic responses that effectively matched the experimental data. The pressure distribution within the models was used to compare the fracture occurrence and location. The spinal levels that sustained vertebral body fracture in the experiment showed higher simulation pressure values in the anterior elements than those in the levels that did not fracture in the reference experiments. Similarly, the spinal levels that sustained posterior element fracture in the experiments showed higher simulation pressure values in the vertebral posterior structures compared to those in the levels that did not sustain fracture. Our study showed that the incorporation of the spinal geometry and orientation could be used to replicate the fracture type and location under dynamic loading. Our results provided an understanding of the lumbar injury mechanisms and knowledge on the load thresholds that could be used for injury prediction with explicit FE lumbar spine models.

Human proximal tubular epithelial cell-derived small extracellular vesicles mediate synchronized tubular ferroptosis in hypoxic kidney injury.

Hypoxia is the key pathobiological trigger of tubular oxidative stress and cell death that drives the transition of acute kidney injury (AKI) to chronic kidney disease (CKD). The mitochondrial-rich proximal tubular epithelial cells (PTEC) are uniquely sensitive to hypoxia and thus, are pivotal in propagating the sustained tubular loss of AKI-to-CKD transition. Here, we examined the role of PTEC-derived small extracellular vesicles (sEV) in propagating the 'wave of tubular death'. Ex vivo patient-derived PTEC were cultured under normoxia (21 % O2) and hypoxia (1 % O2) on Transwell inserts for isolation and analysis of sEV secreted from apical versus basolateral PTEC surfaces. Increased numbers of sEV were secreted from the apical surface of hypoxic PTEC compared with normoxic PTEC. No differences in basolateral sEV numbers were observed between culture conditions. Biological pathway analysis of hypoxic-apical sEV cargo identified distinct miRNAs linked with cellular injury pathways. In functional assays, hypoxic-apical sEV selectively induced ferroptotic cell death (↓glutathione peroxidase-4, ↑lipid peroxidation) in autologous PTEC compared with normoxic-apical sEV. The addition of ferroptosis inhibitors, ferrostatin-1 and baicalein, attenuated PTEC ferroptosis. RNAse A pretreatment of hypoxic-apical sEV also abrogated PTEC ferroptosis, demonstrating a role for sEV RNA in ferroptotic 'wave of death' signalling. In line with these in vitro findings, in situ immunolabelling of diagnostic kidney biopsies from AKI patients with clinical progression to CKD (AKI-to-CKD transition) showed evidence of ferroptosis propagation (increased numbers of ACSL4+ PTEC), while urine-derived sEV (usEV) from these 'AKI-to-CKD transition' patients triggered PTEC ferroptosis (↑lipid peroxidation) in functional studies. Our data establish PTEC-derived apical sEV and their intravesicular RNA as mediators of tubular lipid peroxidation and ferroptosis in hypoxic kidney injury. This concept of how tubular pathology is propagated from the initiating insult into a 'wave of death' provides novel therapeutic check-points for targeting AKI-to-CKD transition.

A Novel Paradigm to Develop Regional Thoracoabdominal Criteria for Behind Armor Blunt Trauma Based on Original Data.

INTRODUCTION: For behind armor blunt trauma (BABT), recent prominent BABT standards for chest plate define a maximum deformation distance of 44 mm in clay. It was developed for soft body armor applications with limited animal, gelatin, and clay tests. The legacy criterion does not account for differing regional thoracoabdominal tolerances to behind armor-induced injury. This study examines the rationale and approaches used in the legacy BABT clay criterion and presents a novel paradigm to develop thoracoabdominal regional injury risk curves. MATERIALS AND METHODS: A review of the original military and law enforcement studies using animals, surrogates, and body armor materials was conducted, and a reanalysis of data was performed. A multiparameter model analysis describes survival-lethality responses using impactor/projectile (mass, diameter, and impact velocity) and specimen (weight and tissue thickness) variables. Binary regression risk curves with ±95% confidence intervals (CIs) and peak deformations from simulant tests are presented. RESULTS: Injury risk curves from 74 goat thorax tests showed that peak deflections of 44.7 mm (±95% CI: 17.6 to 55.4 mm) and 49.9 mm (±95% CI: 24.7 to 60.4 mm) were associated with the 10% and 15% probability of lethal outcomes. 20% gelatin and Roma Plastilina #1 clay were stiffer than goat. The clay was stiffer than 20% gelatin. Penetration diameters showed greater variations (on a test-by-test basis, difference 36-53%) than penetration depths (0-12%) across a range of projectiles and velocities. CONCLUSIONS: While the original authors stressed limitations and the importance of additional tests for refining the 44 mm recommendation, they were not pursued. As live swine tests are effective in developing injury criteria and the responses of different areas of the thoracoabdominal regions are different because of anatomy, structure, and function, a new set of swine and human cadaver tests are necessary to develop scaling relationships. Live swine tests are needed to develop incapacitation/lethal injury risk functions; using scaling relationships, human injury criteria can be developed.

Discovery and validation of serum glycoprotein biomarkers for high grade serous ovarian cancer.

PURPOSE: This study aimed to identify serum glycoprotein biomarkers for early detection of high-grade serous ovarian cancer (HGSOC), the most common and aggressive histotype of ovarian cancer. EXPERIMENTAL DESIGN: The glycoproteomics pipeline lectin magnetic bead array (LeMBA)-mass spectrometry (MS) was used in age-matched case-control serum samples. Clinical samples collected at diagnosis were divided into discovery (n = 30) and validation (n = 98) sets. We also analysed a set of preclinical sera (n = 30) collected prior to HGSOC diagnosis in the UK Collaborative Trial of Ovarian Cancer Screening. RESULTS: A 7-lectin LeMBA-MS/MS discovery screen shortlisted 59 candidate proteins and three lectins. Validation analysis using 3-lectin LeMBA-multiple reaction monitoring (MRM) confirmed elevated A1AT, AACT, CO9, HPT and ITIH3 and reduced A2MG, ALS, IBP3 and PON1 glycoforms in HGSOC. The best performing multimarker signature had 87.7% area under the receiver operating curve, 90.7% specificity and 70.4% sensitivity for distinguishing HGSOC from benign and healthy groups. In the preclinical set, CO9, ITIH3 and A2MG glycoforms were altered in samples collected 11.1 ± 5.1 months prior to HGSOC diagnosis, suggesting potential for early detection. CONCLUSIONS AND CLINICAL RELEVANCE: Our findings provide evidence of candidate early HGSOC serum glycoprotein biomarkers, laying the foundation for further study in larger cohorts.

Similar Concussion Rates in Spring Football and Preseason: Findings From the Concussion Assessment, Research and Education Consortium.

CONTEXT: Increasing attention has been directed toward identifying aspects of football participation for targeted policy change that reduces the concussion risk. Prior researchers evaluated concussion risks during the preseason and regular seasons, leaving the spring season largely unexplored. DESIGN: In this nationally representative observational investigation of 19 National Collegiate Athletic Association Division I collegiate football programs, we assessed concussion rates and head impact exposures during the preseason, regular season, and spring practices from 2014 to 2019. All participating programs recorded the incidence of concussions, and a subset (n = 6) also measured head impact exposures. RESULTS: Analyses by time of year and session type indicated that concussion rates and head impact exposures during all practice sessions and contact practices were higher in the spring and preseason than those in the regular season (P < .05). Concussion rates during the spring season and preseason were statistically similar. CONCLUSIONS: We identified comparable concussion risks in the spring season and preseason, highlighting the need for targeted policy interventions to protect athlete health and safety.

Response of Thoraco-Abdominal Tissue in High-Rate Compression.

Body armor is used to protect the human from penetrating injuries, however, in the process of defeating a projectile, the back face of the armor can deform into the wearer at extremely high rates. This deformation can cause a variety of soft and hard tissue injuries. Finite element modeling (FEM) represents one of the best tools to predict injuries from this high-rate compression mechanism. However, the validity of a model is reliant on accurate material properties for biological tissues. In this study, we measured the stress-strain response of thoraco-abdominal tissue during high-rate compression (1000 and 1900 s-1) using a split Hopkinson pressure bar (SHPB). High-rate material properties of porcine adipose, heart, spleen, and stomach tissue were characterized. At a strain rate of 1000 s-1, adipose (E = 4.7 MPa) had the most compliant stress-strain response, followed by spleen (E = 9.6 MPa), and then heart tissue (E = 13.6 MPa). At a strain rate of 1900 s-1, adipose (E = 7.3 MPa) had the most compliant stress-strain response, followed by spleen (E = 10.7 MPa), heart (E = 14.1 MPa), and stomach (E = 32.6 MPa) tissue. Only adipose tissue demonstrated a consistent rate dependence for these high strain rates, with a stiffer response at 1900 s-1 compared to 1000 s-1. However, comparison of all these tissues to previously published quasi-static and intermediate dynamic experiments revealed a strong rate dependence with increasing stress response from quasi-static to dynamic to high strain rates. Together, these findings can be used to develop a more accurate finite element model of high-rate compression injuries.

Chronicity of repeated blast traumatic brain injury associated increase in oxycodone seeking in rats.

Numerous epidemiological studies have found co-morbidity between non-severe traumatic brain injury (TBI) and substance misuse in both civilian and military populations. Preclinical studies have also identified this relationship for some misused substances. We have previously demonstrated that repeated blast traumatic brain injury (rbTBI) increased oxycodone seeking without increasing oxycodone self-administration, suggesting that the neurological sequelae of traumatic brain injury can elevate opioid misuse liability. Here, we determined the chronicity of this effect by testing different durations of time between injury and oxycodone self-administration and durations of abstinence. We found that the subchronic (four weeks), but not the acute (three days) or chronic (four months) duration between injury and oxycodone self-administration was associated with increased drug seeking and re-acquisition of self-administration following a 10-day abstinence. Examination of other abstinence durations (two days, four weeks, or four months) revealed no effect of rbTBI on drug seeking at any of the abstinence durations tested. Together, these data indicate that there is a window of vulnerability after TBI when oxycodone self-administration is associated with elevated drug seeking and relapse-related behaviors.

Rapid Assessment of Lipidomics Sample Purity and Quantity Using Fourier-Transform Infrared Spectroscopy.

Despite the increasing popularity of liquid chromatography−mass spectrometry (LC-MS)-based lipidomics, there is a lack of accepted and validated methods for lipid extract quality and quantity assessment prior to LC-MS. Fourier-Transform Infrared Spectroscopy (FTIR) has been reported for quantification of pure lipids. However, the impact of complex lipid sample complexity and purity on total lipid quantification accuracy has not been investigated. Here, we report comprehensive assessment of the sample matrix on the accuracy of lipid quantification using Attenuated Total Reflectance (ATR)-FTIR and establish a simple workflow for lipidomics sample quantification. We show that both pure and complex lipids show characteristic FTIR vibrations of CH- and C=O-stretching vibrations, with a quantitative range of 40−3000 ng and a limit of detection of 12 ng, but sample extraction method and local baseline subtraction during FTIR spectral processing significantly impact lipid quantification via CH stretching. To facilitate sample quality screening, we developed the Lipid Quality (LiQ) score from a spectral library of common contaminants, using a ratio of peak heights between CH stretching vibrations maxima and the collective vibrations from amide/amine, CH-stretching minima and sugar moieties. Taking all tested parameters together, we propose a rapid FTIR workflow for routine lipidomics sample quality and quantity assessment and tested this workflow by comparing to the total LC-MS intensity of targeted lipidomics of 107 human plasma lipid extracts. Exclusion of poor-quality samples based on LiQ score improved the correlation between FTIR and LC-MS quantification. The uncertainty of absolute quantification by FTIR was estimated using a 795 ng SPLASH LipidoMix standard to be <10%. With low sample requirement, we anticipate this simple and rapid method will enhance lipidomics workflow by enabling accurate total lipid quantification and normalization of lipid quantity for MS analysis.

Time Delta Head Impact Frequency: An Analysis on Head Impact Exposure in the Lead Up to a Concussion: Findings from the NCAA-DOD Care Consortium.

Sport-related concussions can result from a single high magnitude impact that generates concussive symptoms, repeated subconcussive head impacts aggregating to generate concussive symptoms, or a combined effect from the two mechanisms. The array of symptoms produced by these mechanisms may be clinically interpreted as a sport-related concussion. It was hypothesized that head impact exposure resulting in concussion is influenced by severity, total number, and frequency of subconcussive head impacts. The influence of total number and magnitude of impacts was previously explored, but frequency was investigated to a lesser degree. In this analysis, head impact frequency was investigated over a new metric called 'time delta', the time difference from the first recorded head impact of the day until the concussive impact. Four exposure metrics were analyzed over the time delta to determine whether frequency of head impact exposure was greater for athletes on their concussion date relative to other dates of contact participation. Those metrics included head impact frequency, head impact accrual rate, risk weighted exposure (RWE), and RWE accrual rate. Athletes experienced an elevated median number of impacts, RWE, and RWE accrual rate over the time delta on their concussion date compared to non-injury sessions. This finding suggests elevated frequency of head impact exposure on the concussion date compared to other dates that may precipitate the onset of concussion.

Elevation of fatty acid desaturase 2 in esophageal adenocarcinoma increases polyunsaturated lipids and may exacerbate bile acid-induced DNA damage.

BACKGROUND: The risk of esophageal adenocarcinoma (EAC) is associated with gastro-esophageal reflux disease (GERD) and obesity. Lipid metabolism-targeted therapies decrease the risk of progressing from Barrett's esophagus (BE) to EAC, but the precise lipid metabolic changes and their roles in genotoxicity during EAC development are yet to be established. METHODS: Esophageal biopsies from the normal epithelium (NE), BE, and EAC, were analyzed using concurrent lipidomics and proteomics (n = 30) followed by orthogonal validation on independent samples using RNAseq transcriptomics (n = 22) and immunohistochemistry (IHC, n = 80). The EAC cell line FLO-1 was treated with FADS2 selective inhibitor SC26196, and/or bile acid cocktail, followed by immunofluorescence staining for γH2AX. RESULTS: Metabolism-focused Reactome analysis of the proteomics data revealed enrichment of fatty acid metabolism, ketone body metabolism, and biosynthesis of specialized pro-resolving mediators in EAC pathogenesis. Lipidomics revealed progressive alterations (NE-BE-EAC) in glycerophospholipid synthesis with decreasing triglycerides and increasing phosphatidylcholine and phosphatidylethanolamine, and sphingolipid synthesis with decreasing dihydroceramide and increasing ceramides. Furthermore, a progressive increase in lipids with C20 fatty acids and polyunsaturated lipids with ≥4 double bonds were also observed. Integration with transcriptome data identified candidate enzymes for IHC validation: Δ4-Desaturase, Sphingolipid 1 (DEGS1) which desaturates dihydroceramide to ceramide, and Δ5 and Δ6-Desaturases (fatty acid desaturases, FADS1 and FADS2), responsible for polyunsaturation. All three enzymes showed significant increases from BE through dysplasia to EAC, but transcript levels of DEGS1 were decreased suggesting post-translational regulation. Finally, the FADS2 selective inhibitor SC26196 significantly reduced polyunsaturated lipids with three and four double bonds and reduced bile acid-induced DNA double-strand breaks in FLO-1 cells in vitro. CONCLUSIONS: Integrated multiomics revealed sphingolipid and phospholipid metabolism rewiring during EAC development. FADS2 inhibition and reduction of the high polyunsaturated lipids effectively protected EAC cells from bile acid-induced DNA damage in vitro, potentially through reduced lipid peroxidation.

A Preclinical Rodent Model for Repetitive Subconcussive Head Impact Exposure in Contact Sport Athletes.

Repetitive subconcussive head impact exposure has been associated with clinical and MRI changes in some non-concussed contact sport athletes over the course of a season. However, analysis of human tolerance for repeated head impacts is complicated by concussion and head impact exposure history, genetics, and other personal factors. Therefore, the objective of the current study was to develop a rodent model for repetitive subconcussive head impact exposure that can be used to understand injury mechanisms and tolerance in the human. This study incorporated the Medical College of Wisconsin Rotational Injury Model to expose rats to multiple low-level head accelerations per day over a 4-week period. The peak magnitude of head accelerations were scaled from our prior human studies of contact sport athletes and the number of exposures per day were based on the median (moderate exposure) and 95th percentile (high exposure) number of exposures per day across the human sample. Following the exposure protocol, rats were assessed for cognitive deficits, emotional changes, blood serum levels of axonal injury biomarkers, and histopathological evidence of injury. High exposure rats demonstrated cognitive deficits and evidence of anxiety-like behaviors relative to shams. Moderate exposure rats did not demonstrate either of those behaviors. Similarly, high exposure rats had histopathological evidence of gliosis [i.e., elevated Iba1 intensity and glial fibrillary acidic protein (GFAP) volume relative to shams] in the basolateral amygdala and other areas. Blood serum levels of neurofilament light (NFL) demonstrated a dose response relationship with increasing numbers of low-level head acceleration exposures with a higher week-to-week rate of NFL increase for the high exposure group compared to the moderate exposure group. These findings demonstrate a cumulative effect of repeated low-level head accelerations and provide a model that can be used in future studies to better understand mechanisms and tolerance for brain injury resulting from repeated low-level head accelerations, with scalable biomechanics between the rat and human.

C5b-9 Membrane Attack Complex Formation and Extracellular Vesicle Shedding in Barrett's Esophagus and Esophageal Adenocarcinoma.

The early complement components have emerged as mediators of pro-oncogenic inflammation, classically inferred to cause terminal complement activation, but there are limited data on the activity of terminal complement in cancer. We previously reported elevated serum and tissue C9, the terminal complement component, in esophageal adenocarcinoma (EAC) compared to the precursor condition Barrett's Esophagus (BE) and healthy controls. Here, we investigate the level and cellular fates of the terminal complement complex C5b-9, also known as the membrane attack complex. Punctate C5b-9 staining and diffuse C9 staining was detected in BE and EAC by multiplex immunohistofluorescence without corresponding increase of C9 mRNA transcript. Increased C9 and C5b-9 staining were observed in the sequence normal squamous epithelium, BE, low- and high-grade dysplasia, EAC. C5b-9 positive esophageal cells were morphologically intact, indicative of sublytic or complement-evasion mechanisms. To investigate this at a cellular level, we exposed non-dysplastic BE (BAR-T and CP-A), high-grade dysplastic BE (CP-B and CP-D) and EAC (FLO-1 and OE-33) cell lines to the same sublytic dose of immunopurified human C9 (3 µg/ml) in the presence of C9-depleted human serum. Cellular C5b-9 was visualized by immunofluorescence confocal microscopy. Shed C5b-9 in the form of extracellular vesicles (EV) was measured in collected conditioned medium using recently described microfluidic immunoassay with capture by a mixture of three tetraspanin antibodies (CD9/CD63/CD81) and detection by surface-enhanced Raman scattering (SERS) after EV labelling with C5b-9 or C9 antibody conjugated SERS nanotags. Following C9 exposure, all examined cell lines formed C5b-9, internalized C5b-9, and shed C5b-9+ and C9+ EVs, albeit at varying levels despite receiving the same C9 dose. In conclusion, these results confirm increased esophageal C5b-9 formation during EAC development and demonstrate capability and heterogeneity in C5b-9 formation and shedding in BE and EAC cell lines following sublytic C9 exposure. Future work may explore the molecular mechanisms and pathogenic implications of the shed C5b-9+ EV.

Repeated blast mild traumatic brain injury and oxycodone self-administration produce interactive effects on neuroimaging outcomes.

Traumatic brain injury (TBI) and drug addiction are common comorbidities, but it is unknown if the neurological sequelae of TBI contribute to this relationship. We have previously reported elevated oxycodone seeking after drug self-administration in rats that received repeated blast TBI (rbTBI). TBI and exposure to drugs of abuse can each change structural and functional neuroimaging outcomes, but it is unknown if there are interactive effects of injury and drug exposure. To determine the effects of TBI and oxycodone exposure, we subjected rats to rbTBI and oxycodone self-administration and measured drug seeking and several neuroimaging measures. We found interactive effects of rbTBI and oxycodone on fractional anisotropy (FA) in the nucleus accumbens (NAc) and that FA in the medial prefrontal cortex (mPFC) was correlated with drug seeking. We also found an interactive effect of injury and drug on widespread functional connectivity and regional homogeneity of the blood oxygen level dependent (BOLD) response, and that intra-hemispheric functional connectivity in the infralimbic medial prefrontal cortex positively correlated with drug seeking. In conclusion, rbTBI and oxycodone self-administration had interactive effects on structural and functional magnetic resonance imaging (MRI) measures, and correlational effects were found between some of these measures and drug seeking. These data support the hypothesis that TBI and opioid exposure produce neuroadaptations that contribute to addiction liability.

Association between Preseason/Regular Season Head Impact Exposure and Concussion Incidence in NCAA Football.

PURPOSE: Contact sport athletes are exposed to a unique environment where they sustain repeated head impacts throughout the season and can sustain hundreds of head impacts over a few months. Accordingly, recent studies outlined the role that head impact exposure (HIE) has in concussion biomechanics and in the development of cognitive and brain-based changes. Those studies focused on time-bound effects by quantifying exposure leading up to the concussion, or cognitive changes after a season in which athletes had high HIE. However, HIE may have a more prolonged effect. This study identified associations between HIE and concussion incidence during different periods of the college football fall season. METHODS: This study included 1120 athlete seasons from six National Collegiate Athletic Association Division I football programs across 5 yr. Athletes were instrumented with the Head Impact Telemetry System to record daily HIE. The analysis quantified associations of preseason/regular season/total season concussion incidence with HIE during those periods. RESULTS: Strong associations were identified between HIE and concussion incidence during different periods of the season. Preseason HIE was associated with preseason and total season concussion incidence, and total season HIE was associated with total season concussion incidence. CONCLUSIONS: These findings demonstrate a prolonged effect of HIE on concussion risk, wherein elevated preseason HIE was associated with higher concussion risk both during the preseason and throughout the entire fall season. This investigation is the first to provide evidence supporting the hypothesis of a relationship between elevated HIE during the college football preseason and a sustained decreased tolerance for concussion throughout that season.

Correction: Detecting antimicrobial resistance in Escherichia coli using benchtop attenuated total reflectance-Fourier transform infrared spectroscopy and machine learning.

Correction for 'Detecting antimicrobial resistance in Escherichia coli using benchtop attenuated total reflectance-Fourier transform infrared spectroscopy and machine learning' by Hewa G. S. Wijesinghe et al., Analyst, 2021, DOI: 10.1039/d1an00546d.

Detecting antimicrobial resistance in Escherichia coli using benchtop attenuated total reflectance-Fourier transform infrared spectroscopy and machine learning.

The widespread dissemination of resistance to third-generation cephalosporins in the Enterobacterales through the production of extended-spectrum ß-lactamase (ESBL) is considered a critical global crisis requiring urgent attention of clinicians and scientists alike. Rapid diagnostic methods that can identify microbial resistance profiles closer to the point of care are crucial to minimize the overuse of antimicrobial agents and improve patient outcomes. Although Fourier transform infrared (FTIR) microscopy has shown promise in distinguishing between bacterial species, the high cost and technical requirements of the IR microscope may limit broad clinical use. To address the practical needs of a clinical microbiology laboratory, here, we examine the ability of a lower cost portable benchtop attenuated total reflectance (ATR)-FTIR spectrometer to achieve antimicrobial resistance detection, using a simple, clinically aligned sampling protocol. The technical reproducibility was confirmed through multi-day analysis of an Escherichia coli type strain, which serves as quality control. We generated a dataset of 100 E. coli clinical bloodstream isolates with 63 ceftriaxone resistant blaCTX-M ESBL gene variant strains and developed a classifier for blaCTX-M genotype detection. After assessing 35 machine learning methods using the training set (n = 71), four methods were further optimised, and the best performing method was evaluated using the held-out testing set (n = 29). A tuned support vector machine model with a polynomial kernel, using the 700-1500 cm-1 range achieved a sensitivity of 89.2%, and specificity of 66.7% for detecting blaCTX-M in independent testing, approaching the reported performance of FTIR microscopy. With further algorithm improvement, these data suggest the potential deployment of a portable FTIR spectrometer as a rapid antimicrobial susceptibility prediction platform to enable the efficient use of antimicrobials.