Blast-induced axonal degeneration in the rat cerebellum in the absence of head movement

Blast exposure can injure brain by multiple mechanisms, and injury attributable to direct effects of the blast wave itself have been difficult to distinguish from that caused by rapid head displacement and other secondary processes. To resolve this issue, we used a rat model of blast exposure in which head movement was either strictly prevented or permitted in the lateral plane. Blast was found to produce axonal injury even with strict prevention of head movement. This axonal injury was restricted to the cerebellum, with the exception of injury in visual tracts secondary to ocular trauma. The cerebellar axonal injury was increased in rats in which blast-induced head movement was permitted, but the pattern of injury was unchanged. These findings support the contentions that blast per se, independent of head movement, is sufficient to induce axonal injury, and that axons in cerebellar white matter are particularly vulnerable to direct blast-induced injury.

Repetitive Blast Exposure Increases Appetitive Motivation and Behavioral Inflexibility in Male Mice

Blast exposure (via detonation of high explosives) represents a major potential trauma source for Servicemembers and Veterans, often resulting in mild traumatic brain injury (mTBI). Executive dysfunction (e.g., alterations in memory, deficits in mental flexibility, difficulty with adaptability) is commonly reported by Veterans with a history of blast-related mTBI, leading to impaired daily functioning and decreased quality of life, but underlying mechanisms are not fully understood and have not been well studied in animal models of blast. To investigate potential underlying behavioral mechanisms contributing to deficits in executive functioning post-blast mTBI, here we examined how a history of repetitive blast exposure in male mice affects anxiety/compulsivity-like outcomes and appetitive goal-directed behavior using an established mouse model of blast mTBI. We hypothesized that repetitive blast exposure in male mice would result in anxiety/compulsivity-like outcomes and corresponding performance deficits in operant-based reward learning and behavioral flexibility paradigms. Instead, results demonstrate an increase in reward-seeking and goal-directed behavior and a congruent decrease in behavioral flexibility. We also report chronic adverse behavioral changes related to anxiety, compulsivity, and hyperarousal. In combination, these data suggest that potential deficits in executive function following blast mTBI are at least in part related to enhanced compulsivity/hyperreactivity and behavioral inflexibility and not simply due to a lack of motivation or inability to acquire task parameters, with important implications for subsequent diagnosis and treatment management.

Chronic Anxiety- and Depression-Like Behaviors Are Associated With Glial-Driven Pathology Following Repeated Blast Induced Neurotrauma

Long-term neuropsychiatric impairments have become a growing concern following blast-related traumatic brain injury (bTBI) in active military personnel and Veterans. Neuropsychiatric impairments such as anxiety and depression are common comorbidities that Veterans report months, even years following injury. To understand these chronic behavioral outcomes following blast injury, there is a need to study the link between anxiety, depression, and neuropathology. The hippocampus and motor cortex (MC) have been regions of interest when studying cognitive deficits following blast exposure, but clinical studies of mood disorders such as major depressive disorder (MDD) report that these two regions also play a role in the manifestation of anxiety and depression. With anxiety and depression being common long-term outcomes following bTBI, it is imperative to study how chronic pathological changes within the hippocampus and/or MC due to blast contribute to the development of these psychiatric impairments. In this study, we exposed male rats to a repeated blast overpressure (~17 psi) and evaluated the chronic behavioral and pathological effects on the hippocampus and MC. Results demonstrated that the repeated blast exposure led to depression-like behaviors 36 weeks following injury, and anxiety-like behaviors 2-, and 52-weeks following injury. These behaviors were also correlated with astrocyte pathology (glial-fibrillary acid protein, GFAP) and dendritic alterations (Microtubule-Associated Proteins, MAP2) within the hippocampus and MC regions at 52 weeks. Overall, these findings support the premise that chronic glial pathological changes within the brain contribute to neuropsychiatric impairments following blast exposure.

Transfer Function for Relative Blast Overpressure Through Porcine and Human Skulls In Situ

ABSTRACT Introduction The overarching objective of the Office of Naval Research sponsored Blast Load Assessment Sense and Test (BLAST) program was to quantify neurofunctional risk from repeated blast exposure. However, human studies have limitations in data collection that can only be addressed by animal models. To utilize a large animal model in this work, researchers developed an approach for scaling blast exposure data from animal to human-equivalent loading. For this study, energy interacting with the brain tissue was selected as a translation metric because of the hypothesized association between observed neurological changes and energy transmitted through the skull. This article describes the methodology used to derive an energy-based transfer function capable of serving as a global correspondence rule for primary blast injury exposure, allowing researchers to derive human-appropriate thresholds from animal data. Methods and Materials To generate data for the development of the transfer functions, three disarticulated cadaveric Yucatan minipigs and three postmortem human surrogate heads were exposed to blast overpressure using a large bore, compressed-gas shock tube. Pressure gauges in the free field, on the skull surface, and pressure probes within the brain cavity filled with Sylgard silicone gel recorded the pressure propagation through the skull of each specimen. The frequency components of the freefield and brain cavity measurements from the pig and human surrogates were interrogated in the frequency domain. Doing so quantifies the differences in the amount of energy, in each frequency band, transmitted through both the porcine and the human skull, and the transfer function was calculated to quantify those differences. Results Nonlinear energy transmission was observed for both the porcine and human skulls, indicating that linear scaling would not be appropriate for developing porcine to human transfer functions. This study demonstrated similar responses between species with little to no attenuation at frequencies below 30 Hz. The phase of the pressure transmission to the brain is also similar for both species up to approximately 10 kHz. There were two notable differences between the porcine and human surrogates. First, in the 40-100 Hz range, human subjects have approximately 8 dB more pressure transmitted through the skull relative to porcine subjects. Second, in the 1-10 kHz range, human subjects have up to 10 dB more pressure transmitted into the brain (10 dB more attenuation) relative to the porcine subjects. Conclusions The fundamental goal of this study was to develop pig-to-human transfer functions to allow researchers to interpret data collected from large animal studies and aid in deriving risk functions for repeated blast exposures. Similarities in porcine and human brain physiology make the minipig experimental model an excellent candidate for blast research. However, differences in the skull geometry have historically made the interpretation of animal data difficult for the purposes of characterizing potential neurological risk in humans. Human equivalent loading conditions are critical so that the thresholds are not over- or underpredicted due to differences in porcine skull geometry. This research provides a solution to this challenge, providing a robust methodology for interpreting animal data for blast research.

The Application of Machine Learning to Identify Large Animal Blast Exposure Thresholds

ABSTRACT Introduction The Office of Naval Research sponsored the Blast Load Assessment Sense and Test program to develop a rapid, in-field solution that could be used by team leaders, commanders, and medical personnel to make science-based stand-down decisions for service members exposed to blast overpressure. Toward this goal, the authors propose an ensemble approach based on machine learning (ML) methods to derive a threshold surface for potential neurological deficits that encompasses the intensity of the blast events, the number of exposures, and the period over which the exposures occurred. Because of collection challenges presented by human subjects, the authors utilized data representing a comprehensive set of measures, including structural, behavioral, and cellular changes, from preclinical large animal studies on minipig models. This article describes the development process used to procure the resulting methodology from these studies. Methods and Materials Using an ensemble of ML methods applied to experimental data obtained from 71 Yucatan minipigs, the relationship between blast exposure and neurological deficits was delineated. Despite a relatively small sample size, ML methods with k-fold cross-validation (with k = 5) were justified because of the complexity of the dataset reflecting numerous nonlinear relationships between cellular, structural, and behavioral markers. Based on the physiological responses and environmental measures collected during the large animal study, two models were developed to investigate the relationship between multiple outcome measures and exposure to blast. The histological features model was trained on single-exposure animal data to predict a binary injury response (injured or not) using histological features. The environmental features model related the observed behavioral changes to the environmental parameters collected. Results The histological features model predicted a binary injury outcome from cellular and physiological measurements. Features identified in developing this classification model showed some level of correlation to observed behavioral changes, suggesting that glial activation inflammation and neurodegenerative responses occur even at the lowest levels of blast exposures tested. The results of the environmental features model, which estimated injury risk from environmental blast exposure characteristics, suggested that the observed changes are not just a function of impulse but an average dynamic impulse rate. Noticeable behavioral deficits were observed at loading rates of 100 kPa (impulse/positive duration) or peak pressures of 300-350 kPa, with an approximate positive phase duration of 3.4 ms for single exposure. Based on this analysis, a 3D threshold surface was developed to characterize the potential risk of neurological deficits. Conclusions The ensemble approach facilitated the identification of a pattern of changes across multiple variables to predict the occurrence of changes in brain function. Many changes observed after blast exposure were subtle, making them difficult to measure in human subjects. ML methodologies applied to minipig data demonstrated the value of these techniques in analyzing complex datasets to complement human studies. Importantly, the threshold surface supports the development of science-based blast exposure guidelines.

Development of a Fast-Running Algorithm to Approximate Incident Blast Parameters Using Body-Mounted Sensor Measurements

ABSTRACT Introduction The Office of Naval Research sponsored the Blast Load Assessment-Sense and Test program to develop a rapid, in-field solution that could be used by team leaders, commanders, and medical personnel to make science-based stand-down decisions for service members exposed to blast overpressure. However, a critical challenge to this goal was the reliable interpretation of surface pressure data collected by body-worn blast sensors in both combat and combat training scenarios. Without an appropriate standardized metric, exposures from different blast events cannot be compared and accumulated in a service member’s unique blast exposure profile. In response to these challenges, we developed the Fast Automated Signal Transformation, or FAST, algorithm to automate the processing of large amounts of pressure–time data collected by blast sensors and provide a rapid, reliable approximation of the incident blast parameters without user intervention. This paper describes the performance of the FAST algorithms developed to approximate incident blast metrics from high-explosive sources using only data from body-mounted blast sensors. Methods and Materials Incident pressure was chosen as the standardized output metric because it provides a physiologically relevant estimate of the exposure to blast that can be compared across multiple events. In addition, incident pressure serves as an ideal metric because it is not directionally dependent or affected by the orientation of the operator. The FAST algorithms also preprocess data and automatically flag “not real” traces that might not be from blasts events (false positives). Elimination of any “not real” blast waveforms is essential to avoid skewing the results of subsequent analyses. To evaluate the performance of the FAST algorithms, the FAST results were compared to (1) experimentally measured pressures and (2) results from high-fidelity numerical simulations for three representative real-world events. Results The FAST results were in good agreement with both experimental data and high-fidelity simulations for the three case studies analyzed. The first case study evaluated the performance of FAST with respect to body shielding. The predicted incident pressure by FAST for a surrogate facing the charge, side on to charge, and facing away from the charge was examined. The second case study evaluated the performance of FAST with respect to an irregular charge compared to both pressure probes and results from high-fidelity simulations. The third case study demonstrated the utility of FAST for detonations inside structures where reflections from nearby surfaces can significantly alter the incident pressure. Overall, FAST predictions accounted for the reflections, providing a pressure estimate typically within 20% of the anticipated value. Conclusions This paper presents a standardized approach—the FAST algorithms—to analyze body-mounted blast sensor data. FAST algorithms account for the effects of shock interactions with the body to produce an estimate of incident blast conditions, allowing for direct comparison of individual exposure from different blast events. The continuing development of FAST algorithms will include heavy weapons, providing a singular capability to rapidly interpret body-worn sensor data, and provide standard output for analysis of an individual’s unique blast exposure profile.

Blast Exposure and Self-Reported Hearing Difficulty in Service Members and Veterans Who Have Normal Pure-Tone Hearing Sensitivity: The Mediating Role of Posttraumatic Stress Disorder

Purpose Evidence suggests that military blast exposure may lead to self-reported hearing difficulties despite audiometrically normal hearing. Research identifying potential mechanisms of this association remains limited. The purpose of this article is to evaluate the associations between blast, posttraumatic stress disorder (PTSD), and self-reported hearing difficulty, and to examine PTSD as a possible mediator of the association between blast exposure and hearing difficulty. Method We used baseline data from the Noise Outcomes in Service members Epidemiology (NOISE) study ( n = 477). Participants in this study undergo a comprehensive hearing, and tinnitus if applicable, evaluation and complete a large number of surveys. Pertinent data extracted from these surveys included information on participant's demographics, military service history, including exposure to blast, and health conditions such as symptoms of PTSD. Using regression models and following a formal causal mediation framework, we estimated total associations, natural direct and indirect associations, and percent mediated. Results We found that individuals with blast exposure had higher prevalence of both probable PTSD and self-reported hearing difficulty than individuals who were not blast exposed. Compared with participants without blast exposure, those with blast exposure had twice the prevalence of self-reported hearing difficulty, with 41% of the association mediated through probable PTSD. Conclusion As PTSD is a possible mediator of the association between blast exposure and hearing difficulty, Service members and Veterans with normal pure-tone hearing sensitivity who report hearing difficulties and a history of blast exposure may benefit from evaluation for PTSD symptoms. Supplemental Material https://doi.org/10.23641/asha.16674247

Repetitive Low-level Blast Exposure and Neurocognitive Effects in Army Ranger Mortarmen

ABSTRACT Introduction Occupational exposure to repetitive, low-level blasts in military training and combat has been tied to subconcussive injury and poor health outcomes for service members. Most low-level blast studies to date have focused on explosive breaching and firing heavy weapon systems; however, there is limited research on the repetitive blast exposure and physiological effects that mortarmen experience when firing mortar weapon systems. Motivated by anecdotal symptoms of mortarmen, the purpose of this paper is to characterize this exposure and its resulting neurocognitive effects in order to provide preliminary findings and actionable recommendations to safeguard the health of mortarmen. Materials and Methods In collaboration with the U.S. Army Rangers at Fort Benning, blast exposure, symptoms, and pupillary light reflex were measured during 3 days of firing 81 mm and 120 mm mortars in training. Blast exposure analysis included the examination of the blast overpressure (BOP) and cumulative exposure by mortarman position, as well as comparison to the 4 psi safety threshold. Pupillary light reflex responses were analyzed with linear mixed effects modeling. All neurocognitive results were compared between mortarmen (n = 11) and controls (n = 4) and cross-compared with blast exposure and blast history. Results Nearly 500 rounds were fired during the study, resulting in a high cumulative blast exposure for all mortarmen. While two mortarmen had average BOPs exceeding the 4 psi safety limit (Fig. 2), there was a high prevalence of mTBI-like symptoms among all mortarmen, with over 70% experiencing headaches, ringing in the ears, forgetfulness/poor memory, and taking longer to think during the training week (n ≥ 8/11). Mortarmen also had smaller and slower pupillary light reflex responses relative to controls, with significantly slower dilation velocity (P < 0.05) and constriction velocity (P < 0.10). Conclusion Mortarmen experienced high cumulative blast exposure coinciding with altered neurocognition that is suggestive of blast-related subconcussive injury. These neurocognitive effects occurred even in mortarmen with average BOP below the 4 psi safety threshold. While this study was limited by a small sample size, its results demonstrate a concerning health risk for mortarmen that requires additional study and immediate action. Behavioral changes like ducking and standing farther from the mortar when firing can generally help reduce mortarmen BOP exposure, but we recommend the establishment of daily cumulative safety thresholds and daily firing limits in training to reduce cumulative blast exposure, and ultimately, improve mortarmen’s quality of life and longevity in service.

Tandem Mass Tag-Based Quantitative Proteomic Analysis Reveals Pathways Involved in Brain Injury Induced by Chest Exposure to Shock Waves

Recurrent chest blast exposure can lead to brain inflammation, oxidative stress, and mental disorders in soldiers. However, the mechanism that underlies brain injury caused indirectly by chest blasts remains unclear. It is urgent to find additional reliable biomarkers to reveal the intimate details of the pathogenesis of this phenomenon. We used the term tandem mass tag (TMT) labeling combined with liquid chromatography–tandem mass spectrometry (LC-MS/MS) to screen for differentially expressed proteins in rat brain at different time points after a chest blast. Data are available via ProteomeXchange with the identifier PXD025204. Gene Ontology (GO), the Kyoto Encyclopedia of Genes and Genomes (KEGG), the Database for Annotation, Visualization and Integrated Discovery (DAVID), and Cytoscape analyses were used to analyze the proteomic profiles of blast-exposed rats. In addition, we performed Western blotting to verify protein levels. We identified 6,931 proteins, of which 255 were differentially expressed and 43, 84, 52, 97, and 49 were identified in brain tissues at 12, 24, 48, and 72 h and 1 week after chest blast exposure, respectively. In this study, the GO, KEGG, Clusters of Orthologous Groups of proteins, and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analyses indicated that brain damage caused by chest blast exposure involved many important biological processes and signaling pathways, such as inflammation, cell adhesion, phagocytosis, neuronal and synaptic damage, oxidative stress, and apoptosis. Furthermore, Western blotting confirmed that these differentially expressed proteins and affected signaling pathways were associated with brain damage caused by chest blast exposure. This study identifies potential protein biomarkers of brain damage caused indirectly by chest blast and new targets for the treatment of this condition.