Neurofilament Light Protein: a marker for injury severity, clinical course and outcome following moderate-severe Traumatic Brain Injury

Neurofilament Light Protein: a marker for injury severity, clinical course and outcome following moderate-severe Traumatic Brain Injury | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Neurofilament Light Protein: a marker for injury severity, clinical course and outcome following moderate-severe Traumatic Brain Injury Spyridoula Tsetsou, Leah E McQuillan, David J Barton, John Williamson, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7909088/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 5 You are reading this latest preprint version Abstract Background: Recent studies demonstrate increased Neurofilament Light (NfL) levels across all types of Traumatic Brain Injury (TBI). We aimed to evaluate the relationship between acute temporal NfL with ICU physiology and outcome. Methods: Stored serum from 97 patients with severe TBI were analyzed. Samples werecollected every 6h for the first 24h and then daily through day 10. Temporal NfL patterns were examined using group-based trajectory (TRAJ) modeling, and means were calculated over day 0, 1-5, 6-10 and 0-10. ICU injury burden scores were generated using ridge regression anchored to Glasgow Outcome Scale-Extended (GOSE) over the same time-bins. We used both univariable and multivariable regression to assess NfL TRAJ and means with respect to ICU injury burden, demographic data, clinical factors, and outcome. NfL TRAJ and day 0-10 ICU injury burden were added to a baseline model to evaluate outcome discrimination. Mediation analysis was applied to assess the mediation effect of ICU injury burden between NfL and GOSE. Results: Patients demonstrated increasing levels over the monitoring period. Sixty-three patients had low NfL temporal profile (low-TRAJ), and 34 patients had higher profile (high-TRAJ). Individuals in the high-TRAJ group had lower GCS (p=0.0009), more frequently required barbiturate coma (p=0.01) or decompressive surgery (p=0.02) and had worse 6-months outcome (p=0.001). NfL means and ICU injury burden both negatively correlated with GCS. Multivariate logistic regression showed a 2.24X increased odds of unfavorable GOSE per unit increase in ICU injury burden (p=0.0447). When modeled with ICU injury burden, the NfL high-TRAJ group relationship to outcome was attenuated, with ICU burden being a significant mediator of the NfL relationship to GOSE. Conclusion: Our results suggest that ICU injury burden contributes to serum NfL associations with GOSE. Temporal NfL profiles provide novel information about intracranial pathophysiology as a causal mediator of axonal injury burden following severe TBI. traumatic brain injury critical illness intracranial pressure cerebral perfusion pressure brain tissue oxygenation neurofilament light Figures Figure 1 Figure 2 Figure 3 Introduction Traumatic brain injury (TBI) remains a leading cause of death and major disability, with increasing incidence and health-care costs 1 – 3 . The discovery of blood-based biomarkers, as potential prognostic, monitoring and diagnostic tools, has led to increasing interest and research in this field 4 – 7 . Glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase L1 protein (UCH-L1) levels in blood during the acute phase of TBI, have been approved as an aid in the decision to obtain Computed Tomography (CT) imaging among individuals clinically assessed as having mild TBI 8 , 9 . The neurotrauma field has had a rising interest in exploring the utility of neurofilament light (NfL), a marker showing high sensitivity for axonal injury that maps to subacute/chronic brain injury and MRI brain atrophy after moderate/severe TBI (msTBI) 10 , 11 . However, less is known about the utility this marker may have in reflecting ongoing secondary brain injury among critically ill in the Intensive Care Unit (ICU) with msTBI. Neurofilament (Nf) proteins are neuron-specific intermediate filaments abundant in axons. During axonal injury, they dissociate from the cytoskeleton, cause cellular membrane integrity loss and accumulate in extracellular fluid, cerebrospinal fluid (CSF) and blood. They are highly specific to axonal injury 12 , 13 . NfL, Nf-heavy (NfH) and neurofilament-medium form the three subunits of Nf proteins. NfL is considered to be a biomarker reflecting neurodegeneration and is associated with cognitive impairment in patients with mild cognitive impairment and dementia (Alzheimer’s disease, frontotemporal dementia) 14 – 17 . NfL remains high in CSF and serum following TBI 10 , 11 , 18 and is associated with progressive neurodegeneration in this population 19 . Prior work 20 involving patients with msTBI explored the potential for Nf proteins to predict 6-months cognitive outcome in this patient group. Interestingly, the NfL levels were measured later post-injury (16–90 days post injury) did not correlate with the early NfL levels (0–15 days post injury) or 6-months cognitive performance 20 . However, NfL has been linked with 6- or 12-months functional outcome, in all severities of TBI 10 , 21 . Unlike other biomarkers, blood NfL levels continue to rise up to 6 weeks post msTBI 22 , with the peak NfL level linked to poor 6-months functional outcome and to brain atrophy on MRI 22 . In related work 23 , we recently conducted NfL temporal kinetic profile characterizations from the time of injury through subacute and chronic recovery to find that serum NfL level peak ~ 30 days post-injury; also, both acute and chronic serum NfL levels are associated with global recovery at 6- and 12-months post-injury. We previously screened multiple serum biomarkers, for associations with clinical physiological variables measured over time in the acute injury phase 24 . This work highlighted that GFAP levels in the first 10 days were associated with intracranial pressure (ICP) and cerebral perfusion pressure (CPP) summary variables, injury type presentation on CT scans, mortality and recovery, and demonstrated rapidly declining GFAP levels after initial peak correlated with secondary insults. Also, averaged NfL serum levels in the first 10 days were associated with acute physiological parameters, specifically with brain hypoxia (longer duration of PbtO2 < 10 mmHg) and ICP treatment 24 . However, the role of NfL within the ICU clinical course remains poorly understood. Evidence supporting its utility in ICU-based clinical decision-making is limited, and the potential causal relationship between ICU injury burden profiles and outcomes has not been explored. Thus, we evaluated how acute phase, serial NfL levels correlate with the ICU clinical course and outcomes in patients with msTBI. We hypothesized that NfL levels reflect axonal injury sensitive to the extent of secondary brain injury, as indicated by ICU-associated clinical data, and 6-months global outcomes. Methods Study design and participants We used archived blood samples that were originally collected as part of our clinical trial of erythropoietin (clinicaltrials.gov NCT00313716). Inclusion criteria for this study were: 1) age at least 15 years old, 2) motor GCS ≤ 5 after resuscitation related to a closed TBI and 3) ability to enroll within 6 hours from trauma. Exclusion criteria included 1) patients with GCS of 3 with fixed and dilated pupils, 2) penetrating trauma, 3) pregnancy (positive urine test) and 4) life-threatening injuries or severe pre-existing comorbidities. Patients enrolled in the trial were randomly assigned to treatment with erythropoietin or placebo and to a transfusion threshold of 7 or 10 g/dL using a 2x2 factorial design. Blood samples were collected every 6 hours for the first 24 hours after injury and then once daily through day 10 post-injury. Samples were centrifuged for 15 minutes at 1500g, and serum was removed and stored at -80 o C until analyzed. Patients were offered the option to have research blood samples banked for future studies analyses after clinical trial analysis were completion. Of the 200 trial participants, 114 opted to allow future use of the samples and their associated clinical data. Samples were available on 103 patients but only 97 had at least two sample biomarker measurements available during the biosample collection period, and included in this analysis (Fig. 1 A). Baylor College of Medicine Institutional Review Board approved waived consent for the use of these archived, de-identified blood samples (protocol H-44131). Biomarker assays As described in prior work 24 , we assayed NfL with an ultrasensitive immunoassay using digital array technology (Single Molecule Arrays SiMoA)-based Human Neurology 4-Plex B assay (N4PB; Item 103345). All SiMoA assays were run on the SR-X benchtop assay platform (Quanterix Corp., Lexington MA) at the University of Florida (Gainesville, FL) according to manufacturer’s instructions. The lower limit of quantification (LLOQ), lower limit of detection (LLOD), and dynamic range were 0.625 pg/mL, 0.097 pg/mL, and 0.0971-10,000 pg/mL for NfL, respectively. Inter-assay and intra-assay % CV were 4.6–4.9% and 3.5–7.5%, respectively for NfL. Outcome measures and other ICU variables Demographics and injury severity variables included age, sex, race, GCS sum and motor component (pre- and post-resuscitation), injury severity scale (ISS), clinical exam including pupillary reactivity, prehospital hypotension and hypoxia, and acute CT findings (initial and worst Marshall CT category and a CT burden score). International Mission for Prognosis and Analysis of Clinical Trials in TBI (IMPACT) risk of poor outcome and mortality were also calculated 25 . Glasgow Outcome Scale-Extended (GOSE) was used to determine 6-months outcome. For these analyses, GOSE was dichotomized as favorable (score 5–8) and unfavorable outcome (score 1–4). CT reports for the admission scan were reviewed by neurointensivist and/or neurosurgeon attendings and categorized for pre-specified brain lesion types: intraventricular hemorrhage (IVH), intraparenchymal hemorrhage (IPH), subdural hematoma (SDH), subarachnoid hemorrhage (SAH), epidural hematoma (EDH) or diffuse brain injury (DBI), contusion and midline shift. Details about the acute care, clinical events and treatment were extracted from the electronic medical record (EMR), prepared, and summarized for data analysis. Data including ICP, mean arterial pressure (MAP), CPP, jugular venous oxygen saturation (SjvO2), and brain tissue pO2 (PbtO2) were collected and analyzed. The physiological variables were recorded hourly for as long as the ICP was being monitored over the first 10 days of injury. We used average ICP, duration of time that ICP was higher than 25 mmHg and also higher than 30 mmHg, duration of time of CPP was lower than 50 mmHg, duration of time that PbtO2 was less than 10 mmHg, and duration of time that SjvO2 less than 50%. Average values of other parameters and duration of time of abnormal values were collected as well. Treatments to control ICP (e.g. mannitol, barbiturate coma, decompressive craniectomy) were also collected and analyzed. Sustained increases in ICP were treated according to local protocols and the three-tiered algorithm, including but not limited to sedative medications, CSF drainage, hyperventilation, hyperosmotic therapy, barbiturate induced coma, surgical decompression. Physiological variables data and clinical events were binned into 24-h increments to examine changes over time. ICU injury burden score The ICU injury burden score was calculated as described in our prior work 24 . To calculate the ICU injury burden score, the physiological data were also binned into 24-h increments, and the following conditions known to be associated with poor outcome were quantified: duration of time that ICP > 25 mmHg, CPP < 50 mmHg, and PbtO2 < 10 mmHg; mannitol dose administration, implementation of barbiturate coma and/or decompressive craniectomy. Frequency of out-of-range values was evaluated over the full 10-day time course post-injury and in three separate time epochs: the first 24 h, days 1–5, and days 6–10. Given the inter-related nature of acute care physiology data, and inherent collinearity associated with these multiple ICU factors included in the modeling approach, we generated a composite score reflecting cumulative ICU related physiological injury burden rather than assessing individual factors with respect to biomarker levels and long-term outcome. Generating an aggregate ICU injury burden score through data reduction techniques reduces noise and provides a cumulative measure of relevant injury factors that reflect ICU practice patterns over time. A series of simple logistic regressions were conducted considering ICU injury factors over three-time epochs (first 24h-day 0, days 1–5, and days 6–10) with respect to 6-months GOSE and 6-months mortality. ICU factors associated with outcome with a threshold p-value of < 0.1 were identified using unadjusted linear regression and included in the respective ICU injury burden score derivation. A ridge regression that included each significant ICU factor stated above was then applied to create the ICU injury burden scores for each respective time epoch. This method yielded β-weight estimates for each factor in the model, with smaller β-weights reflecting smaller effect sizes. Appropriate to our research question and analytical approach, this method also penalizes multicollinearity when present, which leads to more reliable β-weight estimates and reduced standard error (SE). This penalty term reduces the impact of multicollinearity by shrinking correlated predictors’ coefficients toward zero in a balanced way, thereby producing more stable and interpretable estimates. The most stable and frequent ridge penalty parameter was selected after iterating a sixfold cross validation 1000 times using the R package glmnet 26 . The coefficients generated were then multiplied to each individual’s set of relevant ICU injury factors and summed to produce the ICU injury burden score in each time epoch. CT burden score An admission CT injury burden score was also generated using ridge regression as described previously 24 . CT reports for the admission scan were reviewed and categorized as mentioned above. The injury types associated with GOSE in univariate logistic regression (midline shift, SAH, SDH, EDH, IPH, IVH, DBI and contusion) were selected for inclusion in the weighted sum reflecting CT injury burden. Group-based trajectory analysis We previously reported temporal profiles for days 0–10 NfL 23 using group-based trajectory (TRAJ) analysis 27 for subjects with ≥ 2 sample timepoints in the first 10 days post-injury. Polynomial orders were assigned to group data from each cohort based on minimizing Bayesian Information Criterion (BIC) and ensuring posterior probability > 80% using CNORM distribution (SAS software, Proc TRAJ). These TRAJ group memberships were leveraged in this study for further analysis of relationships to ICU injury burden score, demographic and clinical factors, and global outcome (GOSE). Statistical analysis Descriptive analyses for continuous variables used mean, standard error (SE) of the mean, median, and interquartile range (IQR). Frequencies and percentages were used to quantify categorical variables. Spearman rank correlation tests were applied to compare continuous variables that were not normally distributed. Univariate and multivariate linear and binary logistic regression models were used, where applicable, to assess associations between NfL means, NfL TRAJ group membership, ICU injury burden score, and demographic and clinical covariates with 6-months outcomes (GOSE). ICU injury burden score was evaluated, using the Baron and Kenny approach 28 for a possible mediating relationship between NfL mean levels—and also TRAJ group membership—and 6-month outcome. The area under the receiver operating characteristic curve (AUC) was used to evaluate the discriminative performance of each model. DeLong’s test was employed to compare the AUCs between models, assessing whether differences in model performance were significant. Multivariate models assessing ICU injury burden as a mediator were adjusted for age and sex. Results were reported when the total effect was greater than the direct effect and a p-value threshold less than 0.1 was met. Mediation percentage was calculated as the proportion of the indirect effect to the total effect, shown below. Statistical analyses were conducted using SAS Version 9.4 (Cary, North Carolina), R and GraphPad Prism Version 10 (Boston, Massachusetts). For all analyses, a p-value ≤ 0.05 was considered statistically significant. Results Demographics, clinical, ICU variables and outcomes by NfL TRAJ Ninety-seven (97) patients had samples and data available and were used for analysis. TRAJ analysis identified two distinct subgroups with high (n=34) vs low (n=63) NfL levels; both groups exhibited increasing levels over the 10-day monitoring period (Figure 1B ). Table 1A shows the demographics, clinical and radiological characteristics of the two groups. The high TRAJ group had worse admission GCS (p=0.0009), higher CT burden scores (p=0.0086), more frequently compressed basal cisterns on admission brain CT (p=0.03), and prehospital hypotension (p=0.046). Higher IMPACT score for risk of poor outcome and risk of mortality was also observed for those in high TRAJ group (p<0.0001, p <0.0001 respectively). Similarly, high TRAJ group membership was associated with an increased mortality rate (p=0.001) and worse clinical outcome with GOSE assessment at 6 months (p=0.001). Table 1B shows the ICU physiological variables for patients of the two TRAJ groups. Average ICP was higher for in the high TRAJ group (p=0.01), and this group had also longer duration of time of ICP >25mmHg (p=0.01) and >30mmHg (p=0.0069). High TRAJ group members also was associated with more prolonged cerebral hypoperfusion (7.58hrs vs 2.88hrs, p=0.01), though with only trend-level differences observed in the average CPP value. Patients in the high TRAJ group more frequently required barbiturate coma (p=0.01) or decompressive surgery (p=0.02) for controlling ICP. Cross-sectional NfL and ICU injury burden relationships Day 0, Day 1-5, Day 6-10, and Day 0-10 means for CSF and serum NfL were all screened against their respective ICU injury burden by time epoch (not shown). Serum NfL, but not CSF, levels were associated with ICU injury burden at all time points. Figure 2A presents a cross-leg framework of univariate linear regression models that show the consistent interrelatedness between serum NfL and ICU injury burden factors over the first 10 days post-injury. Day 0 NfL means were significantly associated with Day 1-5 NfL means, and subsequently Day 1-5 NfL means to Day 6-10 NfL means (p<0.001 both comparisons). ICU injury burden demonstrated a similar relationship between adjacent time epochs (p<0.001 both comparisons). Additionally, mean NfL and ICU injury burden scores were significantly related to each other within time epoch (Day 0 NfL to Day 0 ICU injury burden) and across time epochs (day 0 to Day 1-5, and Day 1-5 to Day 6-10). NfL, ICU injury burden, demographic and clinical factors relationship A correlational heatmap ( Figure 2B ) presents pairwise comparisons (by Spearman rank correlation) between NfL means and ICU injury burden scores at all time epochs (Days: 0, 1-5, 6-10, 0-10) with GCS, age and CT burden. Each cell shows the correlation coefficient (r) for the pairwise comparison. Mean NfL levels were positively associated with ICU injury burden and CT burden, and negatively associated with GCS. ICU injury burden was also negatively associated with GCS and positively associated with CT burden at all time epochs. Age had a stronger relationship to ICU injury burden at Day 0 and 1-5 than later timepoints. NfL means and ICU injury burden scores were strongly associated levels and scores at other time epochs. NfL means at all time points were associated, in univariate regression, with 6-months GOSE, with higher NfL levels being associated with unfavorable outcome ( Table 2A ). When adjusting for the ICU injury burden at the corresponding time point ( Table 2B ), the NfL mean relationships to outcome were variably attenuated, while ICU injury burden scores were significantly related to outcome (p<0.05 all comparisons). Here, Day 0 mean NfL was no longer significant in relationship to GOSE when Day 0 ICU injury burden was introduced (p=0.2497); however, Day 1-5 and Day 6-10 remained significant (p=0.0362 and p=0.0223, respectively). Together, the discriminative performance improved when ICU injury burden score was added to outcome models that included mean NfL levels. Each time epoch model showed a significant improvement in AUC (NfL only model vs. NfL + ICU injury burden model) when comparing ROC curves using Delong’s Test (p<0.05 all comparisons). In preparation for multivariate model development, demographic (age, sex) and clinical injury factors (ER GCS, best in 24h GCS, CT burden) were investigated for NfL TRAJ and mean NfL measured over different time bins, with ICU injury burden and global outcome modeled as dependent variables ( Table 3 ). Older age was associated with higher Day 0 ICU injury burden and increased odds of unfavorable outcome. Injury factors (ER and best in 24h GCS, CT burden) were associated with both NfL TRAJ and unfavorable outcomes. Those with more severe injuries had an increased odds of having high NfL TRAJ profiles over the first 10 days post-injury and unfavorable 6-months outcome. CT burden was the factor most strongly associated with all ICU injury burden time epochs (p<0.0001 all comparisons). NfL relationship to outcome in the context of ICU clinical course and demographic factors Multivariate logistic regression models were developed to examine relationships between the pattern of serum NfL levels over time and outcome, after adjusting for other injury severity variables known to be associated with outcome (age, sex, best in 24hr GCS and CT burden score). The baseline characteristic model is shown in Table 4A, with NfL TRAJ and ICU injury burden added incrementally to demonstrate the relative influence of model factors and the change in outcome discrimination by AUC (from 0.787 to 0.803 and 0.819, respectively). Here, lower GCS (p=0.0104) and greater CT burden (p=0.004) were associated with increased odds of unfavorable GOSE ( Table 4A ). When adding Day 0-10 NfL TRAJ to the model, these relationships were attenuated (GCS: p=0.0241, CT burden score: p=0.004) yet remained significant. There was a trend level association with high NfL TRAJ membership being associated with 2.57 increased odds of unfavorable GOSE versus the low TRAJ membership ( Table 4B ). Lastly, when factoring in Day 0-10 ICU burden into the model ( Table 4C ), the CT burden and NfL TRAJ relationships were fully attenuated (p>0.1), and the GCS association with outcome was attenuated to a lesser degree, remaining significantly associated with outcome (p=0.0106). For every unit increase in Day 0-10 ICU burden, there was 2.24 increased odds of unfavorable GOSE (p=0.0447). The AUC increased with the addition of NfL TRAJ and ICU injury burden; however, pairwise comparisons of ROC curves using DeLong’s test revealed no statistically significant differences in AUC between models (all p-values > 0.23), indicating similar discriminative performance. The dynamic role of ICU burden as a mediator of NfL associations over time with 6-months outcome was tested ( Figure 3 ) while adjusting for age and sex as covariates. The mediating effects of ICU burden on day 0 mean and day 1-5 mean NfL relationship to GOSE scores were tested subsequent ICU burden time epochs ( Figure 3A ), and Day 6-10 ICU burden was assessed as a mediator for the day 1-5 NfL mean association with outcome ( Figure 3B ). Day 1-5 ICU injury burden mediated 28% of the day 0 NfL mean association with unfavorable GOSE. Similarly, Day 6-10 ICU burden mediated 39.9% of the day 1-5 NfL mean association with unfavorable GOSE; however, the direct effect remained trend-level (p=0.0861). Figure 3C demonstrates Day 0-10 ICU injury burden as a mediator of the NfL TRAJ relationship to GOSE. Here, the overall burden score mediated 28.9% of the NfL TRAJ association with GOSE resulting in a trend-level direct effect (p=0.0501). Together these models suggest that ICU injury burden is a partial mediator of the NfL biomarker associations with global outcome. Discussion This study highlights temporal and overall associations between NfL and ICU physiological parameters over the first 10 days after msTBI, and their collective impact on global outcome, while considering initial brain injury. The main strengths of the study include the serial biomarker measurements over the first 10 days post-injury along with the detailed clinical, ICU and radiological variables available for analyses. Previous studies have examined serial NfL measurements post-TBI, correlating increasing NfL serum or CSF levels over time with worse clinical outcome. Nimer et al 29 studied 182 patients with all TBI severities and collected 1 to 3 samples per patient over the course of 2 weeks. NfL levels showed an upward trend over time and correlated to outcome. Shahim et al 10 , evaluated 72 severe TBI patients where daily NfL measurements were obtained from day 0 to 12 post-injury. NfL levels were increased at admission, continued to rise daily, and also were associated with clinical outcome. In a longer time-course study we showed that serum NfL levels rise from the acute phase and peak at 30 days post-injury and both acute serum and CSF NfL levels discriminated 6- and 12-months outcome 23 . While that study data included a portion of this cohort 23 , we similarly highlight that serum NfL levels keep rising in the acute phase post-TBI. This linear increase in NfL likely reflects ongoing axonal injury after the initial insult and the importance of serial measurements assessing longitudinal biomarker patterns rather than single time point measurement strategies. The TRAJ analysis approach 27 demonstrated temporal NfL dynamics by identifying two groups of individuals with unique serum NfL pattern over the first 10 days post-injury. A major advantage of TRAJ is that it is a clustering algorithm for assessing longitudinal patterns rather than single-point, averaged, or peak biomarker level for diagnostic or prognostic models. Its capacity for patient clustering for potential TBI patient subtype identification has been demonstrated for a variety of CNS derived 23 , 30 – 33 and systemic biomarkers 34 – 38 . Validation studies are required to address the minimum time needed for patient assigned to a TRAJ group in real time based on their initial NfL levels; however, this methodology, and the dynamic relationships between NFL and ICU burden, shows potential utility for early clinical and prognostic management. NfL levels demonstrated a linear increase over the first 10 days post-injury, contrary to GFAP, which presented with an early peak followed by variable decay patterns over the acute period 24 . Rising NfL elevations over time likely reflect higher degree of evolving secondary axonal injury, as NfL means are closely associated with ICU injury burden over the full 10-day time course post-injury. The regression-based cross lag analysis showed mean NfL and ICU injury burden scores were significantly related to each other within time epoch from day 0 (Day 0 NfL to Day 0 ICU injury burden) and across subsequent time epochs (day 0 to Day 1–5, and Day 1–5 to Day 6–10). In contrast, Day 0 GFAP wasn’t associated with Day 0 ICU injury burden when using a similar analysis construct, highlighting the strong correlation of NfL with initial injury severity. NfL levels were strongly correlated with GCS and CT burden. NfL means also positively correlated with both initial GCS, and best in 24h GCS, across all time epochs, with CT burden scores primarily associated with day 0 and day 1–5 NfL levels. Similarly, higher NfL TRAJ groups were significantly associated with worse GCS and CT burden. This contrasts with the GFAP analysis 24 , where the CT characteristics but not the initial GCS and baseline injury severity factors were associated to the GFAP TRAJ group. Notably, other studies have shown GFAP levels within the first 24 hours correlated with admission GCS 39 . Both GFAP and NfL rise with aging in the general population 40 , 41 , 42 . We found no age differences between low and high TRAJ groups. This work contrasts with our GFAP findings, where individuals in the high TRAJ GFAP group were significantly older 24 . To further evaluate the NfL association with ICU factors and outcome, age and sex were introduced as covariates in mediation analysis. GCS and CT burden scores were excluded due to associations between NfL TRAJ, ICU injury burden and GOSE, allowing NfL to be treated as the biological readout of initial injury severity and allowing it to be effectively modeled as a temporally dynamic variable in pathway analysis without the absence of confounding covariates with shared variance. Mediation analyses suggested that the ICU injury burden is a partial mediator, indicating that NfL biomarker associations with functional outcome occur via secondary CNS physiological injury response in response to neurocritical illness. Taken together, these data suggest NfL is a robust blood-based biomarker representation of initial neurological injury severity, and interestingly, tracks closely with ICU factor time course. Independent of these correlative factors, higher NfL TRAJ and mean NfL levels measurement over the 10-day time epochs were each significantly associated with worse outcome. When introducing ICU injury burden into the modeling framework, these relationships were attenuated, suggesting ICU burden is in the causal pathway explaining a significant amount of the relationship between acute NfL levels and global outcome. These findings align with prior studies that have confirmed elevated NfL after a TBI of any severity is associated with degree of axonal injury 11 , 43 , and thus worse outcome at 6 months; however, the fact that acute ICU appears to have a causal effect on outcome suggests that there may be modifiable targets within ICU care best practice pathways where serial NfL monitoring may be a useful adjunct to clinical management and assessing clinical treatment effectiveness. To the best of our knowledge this is the first time NfL has been linked with acute ICU physiological parameters and acute treatment. Patients in the high TRAJ group had more frequent refractory intracranial hypertension and cerebral hypoperfusion requiring advanced and invasive treatments. These findings build on previous work linking NfL to axonal injury and outcome post-TBI by providing an additional lens through ICU-related physiology to present opportunities for acute management and treatment. Limitations Our study has some limitations. Our sample size is relatively small, representing a single Level 1 trauma center’s experience. The clinical information available was exported from EMR and collected prospectively as part of a previous treatment trial. We did not assess the impact of each ICP treatment component on NfL levels, as in clinical practice multiple treatment measures are done simultaneously. Future larger, multicenter trials are necessary to validate the NfL role in acute ICU setting. Samples were only collected daily up to day 10, with no chronic samples available. However, for the purposes of this study, having serial samples in the first 10 days of ICU stay was sufficient to show the importance of longitudinal biomarker patterns in their relationship with ICU metrics over time. This work presents novel approaches to handling CT injury profiles and longitudinal ICU factors in time-specific load score derivations. However, the granular time course data and the presence of multicollinearity among NfL, clinical factors, and outcome limit the effectiveness of traditional regression approaches and may necessitate more sophisticated analytical techniques (e.g. machine learning) in larger cohorts to establish NFL as a biomarker for real-time clinical decision making. Future research with more frequent serial measurements over a longer time course, with multiple biomarker inclusion and advanced analysis planning, would further refine the role of NfL in individualized ICU care and prognostication, and provide treatment trial readouts for monitoring effectiveness. Conclusion Serum NfL levels correlate with worse clinical outcomes after msTBI and are associated with ICU clinical course. The linear increase of NfL levels and strong correlation with ICU burden and 6-months outcome provide information about the association with secondary brain injury following msTBI. Future research using a high-resolution sampling approach in assessing extended longitudinal patterns, as well as multi-marker panels for treatment triaging and clinical outcome is necessary in preparation for large scale multisite trials focused on assessing complex biological patterns and relationships to CNS physiology co-occurring critical illness associated with msTBI. Declarations Authors contributions : Conceptualization: ST, CR, LM, KKW, AKW Data collection: CR, GC, KKW Data visualization, analysis, and interpretation: ST, LM, CR, DB, KKW, AKW Manuscript writing: ST, LM, CR, AKW Manuscript review and critical revision: CR, DB, JW, DL, FK, GC, RR, KKW, AKW Funding: CR, JW, RR, KKW AKW All coauthors have seen and agree with the contents of the manuscript; the manuscript complies with all instructions to authors; requirements for authorship have been met; each author believes that the manuscript represents honest work; the STROBE checklist was used. This work has not been published elsewhere and is not under consideration by another journal and is not redundant with previous work. We used archived blood samples that were originally collected as part of our clinical trial of erythropoietin (clinicaltrials.gov NCT00313716). Patients were offered the option to have research blood samples banked for future studies analyses after clinical trial analysis were completion. Baylor College of Medicine Institutional Review Board approved waived consent for the use of these archived, de-identified blood samples (protocol H-44131). Funding: This project was funded by Department of Defense grant W81XWH-19-2-0012 (samples analysis, data analysis) and Department of Defense grant Gap-Based Milieu Biomarkers for Traumatic Brain Injury (GAMBIT- TBI) W81XWH2211089 (data analysis, manuscript preparation). The clinical trial of erythropoietin that originally collected the samples used in this study was pre-registered at clinicaltrials.gov (NCT00313716). Further support received from VA research grant IK2RX002490 and the Brain Rehabilitation Research Center, Malcom Randall VAMCGainesville FL. Disclosures : KKW is a share-holder of Gryphon Bio, Inc. The other authors have no relevant disclosures related to this work. References Taylor CA, Bell JM, Breiding MJ, Xu L. Traumatic Brain Injury-Related Emergency Department Visits, Hospitalizations, and Deaths - United States, 2007 and 2013. MMWR Surveill Summ 2017;66(9):1-16. DOI: 10.15585/mmwr.ss6609a1. Hyder AA, Wunderlich CA, Puvanachandra P, Gururaj G, Kobusingye OC. The impact of traumatic brain injuries: a global perspective. NeuroRehabilitation 2007;22(5):341-53. (https://www.ncbi.nlm.nih.gov/pubmed/18162698). Maas AIR, Menon DK, Manley GT, et al. Traumatic brain injury: progress and challenges in prevention, clinical care, and research. Lancet Neurol 2022;21(11):1004-1060. DOI: 10.1016/S1474-4422(22)00309-X. Shahim P, Politis A, van der Merwe A, et al. Time course and diagnostic utility of NfL, tau, GFAP, and UCH-L1 in subacute and chronic TBI. Neurology 2020;95(6):e623-e636. DOI: 10.1212/WNL.0000000000009985. Mouhieddine TH, El Houjeiri L, Sabra M, Hayes RL, Mondello S. CNS Trauma Biomarkers and Surrogate Endpoints Pipeline from Bench to Bedside: A Translational Perspective. In: Kobeissy FH, ed. Brain Neurotrauma: Molecular, Neuropsychological, and Rehabilitation Aspects. Boca Raton (FL)2015. Mondello S, Hayes RL. Biomarkers. Handb Clin Neurol 2015;127:245-65. DOI: 10.1016/B978-0-444-52892-6.00016-7. Nielson JL, Cooper SR, Yue JK, et al. Uncovering precision phenotype-biomarker associations in traumatic brain injury using topological data analysis. PLoS One 2017;12(3):e0169490. DOI: 10.1371/journal.pone.0169490. Korley FK, Jain S, Sun X, et al. Prognostic value of day-of-injury plasma GFAP and UCH-L1 concentrations for predicting functional recovery after traumatic brain injury in patients from the US TRACK-TBI cohort: an observational cohort study. Lancet Neurol 2022;21(9):803-813. DOI: 10.1016/S1474-4422(22)00256-3. Bazarian JJ, Biberthaler P, Welch RD, et al. Serum GFAP and UCH-L1 for prediction of absence of intracranial injuries on head CT (ALERT-TBI): a multicentre observational study. Lancet Neurol 2018;17(9):782-789. DOI: 10.1016/S1474-4422(18)30231-X. Shahim P, Politis A, van der Merwe A, et al. Neurofilament light as a biomarker in traumatic brain injury. Neurology 2020;95(6):e610-e622. DOI: 10.1212/WNL.0000000000009983. Gao W, Zhang Z, Lv X, et al. Neurofilament light chain level in traumatic brain injury: A system review and meta-analysis. Medicine (Baltimore) 2020;99(38):e22363. DOI: 10.1097/MD.0000000000022363. Yuan A, Rao MV, Veeranna, Nixon RA. Neurofilaments and Neurofilament Proteins in Health and Disease. Cold Spring Harb Perspect Biol 2017;9(4). DOI: 10.1101/cshperspect.a018309. Gaetani L, Blennow K, Calabresi P, Di Filippo M, Parnetti L, Zetterberg H. Neurofilament light chain as a biomarker in neurological disorders. J Neurol Neurosurg Psychiatry 2019;90(8):870-881. DOI: 10.1136/jnnp-2018-320106. Mattsson N, Cullen NC, Andreasson U, Zetterberg H, Blennow K. Association Between Longitudinal Plasma Neurofilament Light and Neurodegeneration in Patients With Alzheimer Disease. JAMA Neurol 2019;76(7):791-799. DOI: 10.1001/jamaneurol.2019.0765. Benedet AL, Ashton NJ, Pascoal TA, et al. Plasma neurofilament light associates with Alzheimer's disease metabolic decline in amyloid-positive individuals. Alzheimers Dement (Amst) 2019;11:679-689. DOI: 10.1016/j.dadm.2019.08.002. Rohrer JD, Woollacott IO, Dick KM, et al. Serum neurofilament light chain protein is a measure of disease intensity in frontotemporal dementia. Neurology 2016;87(13):1329-36. DOI: 10.1212/WNL.0000000000003154. Olsson B, Lautner R, Andreasson U, et al. CSF and blood biomarkers for the diagnosis of Alzheimer's disease: a systematic review and meta-analysis. Lancet Neurol 2016;15(7):673-684. DOI: 10.1016/S1474-4422(16)00070-3. Ljungqvist J, Zetterberg H, Mitsis M, Blennow K, Skoglund T. Serum Neurofilament Light Protein as a Marker for Diffuse Axonal Injury: Results from a Case Series Study. J Neurotrauma 2017;34(5):1124-1127. DOI: 10.1089/neu.2016.4496. Shahim P, Pham DL, van der Merwe AJ, et al. Serum NfL and GFAP as biomarkers of progressive neurodegeneration in TBI. Alzheimers Dement 2024;20(7):4663-4676. DOI: 10.1002/alz.13898. Trifilio E, Bottari S, McQuillan LE, et al. Temporal Profile of Serum Neurofilament Light (NF-L) and Heavy (pNF-H) Level Associations With 6-Month Cognitive Performance in Patients With Moderate-Severe Traumatic Brain Injury. J Head Trauma Rehabil 2024. DOI: 10.1097/HTR.0000000000000932. Shahim P, Gren M, Liman V, et al. Serum neurofilament light protein predicts clinical outcome in traumatic brain injury. Sci Rep 2016;6:36791. DOI: 10.1038/srep36791. Graham NSN, Zimmerman KA, Moro F, et al. Axonal marker neurofilament light predicts long-term outcomes and progressive neurodegeneration after traumatic brain injury. Sci Transl Med 2021;13(613):eabg9922. DOI: 10.1126/scitranslmed.abg9922. Wang KKW, Barton DJ, McQuillan LE, et al. Parallel Cerebrospinal Fluid and Serum Temporal Profile Assessment of Axonal Injury Biomarkers Neurofilament-Light Chain and Phosphorylated Neurofilament-Heavy Chain: Associations With Patient Outcome in Moderate-Severe Traumatic Brain Injury. J Neurotrauma 2024;41(13-14):1609-1627. DOI: 10.1089/neu.2023.0449. Robertson CS, Martinez FS, McQuillan LE, et al. Serial Measurements of Serum Glial Fibrillary Acidic Protein in Moderate-Severe Traumatic Brain Injury: Potential Utility in Providing Insights into Secondary Insults and Long-Term Outcome. J Neurotrauma 2024;41(1-2):73-90. DOI: 10.1089/neu.2023.0111. Steyerberg EW, Mushkudiani N, Perel P, et al. Predicting outcome after traumatic brain injury: development and international validation of prognostic scores based on admission characteristics. PLoS Med 2008;5(8):e165; discussion e165. DOI: 10.1371/journal.pmed.0050165. Friedman J, Hastie T, Tibshirani R. Regularization Paths for Generalized Linear Models via Coordinate Descent. J Stat Softw 2010;33(1):1-22. (https://www.ncbi.nlm.nih.gov/pubmed/20808728). Niyonkuru C, Wagner AK, Ozawa H, Amin K, Goyal A, Fabio A. Group-based trajectory analysis applications for prognostic biomarker model development in severe TBI: a practical example. J Neurotrauma 2013;30(11):938-45. DOI: 10.1089/neu.2012.2578. Baron RM, Kenny DA. The moderator-mediator variable distinction in social psychological research: conceptual, strategic, and statistical considerations. J Pers Soc Psychol 1986;51(6):1173-82. DOI: 10.1037//0022-3514.51.6.1173. Al Nimer F, Thelin E, Nystrom H, et al. Comparative Assessment of the Prognostic Value of Biomarkers in Traumatic Brain Injury Reveals an Independent Role for Serum Levels of Neurofilament Light. PLoS One 2015;10(7):e0132177. DOI: 10.1371/journal.pone.0132177. Rubenstein R, McQuillan L, Wang KKW, et al. Temporal Profiles of P-Tau, T-Tau, and P-Tau:Tau Ratios in Cerebrospinal Fluid and Blood from Moderate-Severe Traumatic Brain Injury Patients and Relationship to 6-12 Month Global Outcomes. J Neurotrauma 2024;41(3-4):369-392. DOI: 10.1089/neu.2022.0479. Kumar RG, Diamond ML, Boles JA, et al. Acute CSF interleukin-6 trajectories after TBI: associations with neuroinflammation, polytrauma, and outcome. Brain Behav Immun 2015;45:253-62. DOI: 10.1016/j.bbi.2014.12.021. Wagner AK, Amin KB, Niyonkuru C, et al. CSF Bcl-2 and cytochrome C temporal profiles in outcome prediction for adults with severe TBI. J Cereb Blood Flow Metab 2011;31(9):1886-96. DOI: 10.1038/jcbfm.2011.31. Goyal A, Failla MD, Niyonkuru C, et al. S100b as a prognostic biomarker in outcome prediction for patients with severe traumatic brain injury. J Neurotrauma 2013;30(11):946-57. DOI: 10.1089/neu.2012.2579. Munoz MJ, Kumar RG, Oh BM, et al. Cerebrospinal Fluid Cortisol Mediates Brain-Derived Neurotrophic Factor Relationships to Mortality after Severe TBI: A Prospective Cohort Study. Front Mol Neurosci 2017;10:44. DOI: 10.3389/fnmol.2017.00044. Santarsieri M, Kumar RG, Kochanek PM, Berga S, Wagner AK. Variable neuroendocrine-immune dysfunction in individuals with unfavorable outcome after severe traumatic brain injury. Brain Behav Immun 2015;45:15-27. DOI: 10.1016/j.bbi.2014.09.003. Santarsieri M, Niyonkuru C, McCullough EH, et al. Cerebrospinal fluid cortisol and progesterone profiles and outcomes prognostication after severe traumatic brain injury. J Neurotrauma 2014;31(8):699-712. DOI: 10.1089/neu.2013.3177. Barton DJ, Kumar RG, Schuster AA, Juengst SB, Oh BM, Wagner AK. Acute Cortisol Profile Associations With Cognitive Impairment After Severe Traumatic Brain Injury. Neurorehabil Neural Repair 2021;35(12):1088-1099. DOI: 10.1177/15459683211048771. Wagner AK, McCullough EH, Niyonkuru C, et al. Acute serum hormone levels: characterization and prognosis after severe traumatic brain injury. J Neurotrauma 2011;28(6):871-88. DOI: 10.1089/neu.2010.1586. Okonkwo DO, Puffer RC, Puccio AM, et al. Point-of-Care Platform Blood Biomarker Testing of Glial Fibrillary Acidic Protein versus S100 Calcium-Binding Protein B for Prediction of Traumatic Brain Injuries: A Transforming Research and Clinical Knowledge in Traumatic Brain Injury Study. J Neurotrauma 2020;37(23):2460-2467. DOI: 10.1089/neu.2020.7140. Khalil M, Pirpamer L, Hofer E, et al. Serum neurofilament light levels in normal aging and their association with morphologic brain changes. Nat Commun 2020;11(1):812. DOI: 10.1038/s41467-020-14612-6. Tybirk L, Hviid CVB, Knudsen CS, Parkner T. Serum GFAP - reference interval and preanalytical properties in Danish adults. Clin Chem Lab Med 2022;60(11):1830-1838. DOI: 10.1515/cclm-2022-0646. Disanto G, Barro C, Benkert P, et al. Serum Neurofilament light: A biomarker of neuronal damage in multiple sclerosis. Ann Neurol 2017;81(6):857-870. DOI: 10.1002/ana.24954. Shahim P, Zetterberg H, Tegner Y, Blennow K. Serum neurofilament light as a biomarker for mild traumatic brain injury in contact sports. Neurology 2017;88(19):1788-1794. DOI: 10.1212/WNL.0000000000003912. Tables Table 1A: NfL trajectory group comparisons Serum NfL TRAJ GROUP Low NfL High NfL p-value Number of patients: 63 34 Sex - Male - Female - 51 - 12 - 28 - 6 0.86 Age (mean) - 32.3 34.6 0.6884 Admission Motor GCS - 1-3 - 4-6 - 17 (27%) - 46 (73%) - 22 (64%) - 12 (36%) 0.0005 Admission Sum GCS - 3-5 - 6-8 - >8 - 15 (24%) - 30 (48%) - 18 (28%) - 21 (61%) - 7 (21%) - 6 (18%) 0.0009 GCS (best in 24h), median [IQR] - 6 [4-7] - 4.5 [4-7] 0.0646 Pupils: - Both reactive - One reactive - Neither reactive - 46 (73%) - 6 (10%) - 11 (17%) - 17 (50%) - 6 (18%) - 11 (32%) 0.76 Pre hospital hypotension - Absent - Present - 59 (94%) - 4 (6%) - 27 (80%) - 7 (20%) 0.046 Pre hospital Hypoxia - Absent - Present - 51 (81%) - 12 (19%) - 22 (65%) - 12 (35%) 0.08 Admission Marshall CT - Diffuse injury 2 - Diffuse injury 3 - Evacuated Mass Lesion - Unevacuated Mass Lesion - 31 - 14 - 16 - 2 - 8 - 13 - 12 - 1 0.09 Subarachnoid Hemorrhage - Present - Absent - 41 (65%) - 22 (35%) - 27 (79%) - 7 (21%) 0.16 Basal Cisterns - Normal - Compressed/absent - 37 (59%) - 26 (41%) - 12 (35%) - 22 (65%) 0.03 Midline shift - None - 15 mm - 44 (70%) - 8 (13%) - 9 (14%) - 2 (3%) - 16 (47%) - 6 (18%) - 10 (29%) - 2 (6%) 0.15 Diffuse Brain Injury (DBI) - Present - Absent - 45 (71.4%) - 18 (28.6%) - 22 (64.7%) - 12 (35.3%) 0.4943 CT Burden, median [IQR] 0.51 [0.5-1.62] 1.62 [0.87-1.97] 0.0086 Ventricles - Normal - Small - Enlarged - 29 (46%) - 33 (52%) - 1 (2%) - 12 (35%) - 22 (65%) - 0 (0%) 0.53 IMPACT risk of poor outcome (mean) 0.33 0.58 <0.0001 IMPACT risk of mortality (mean) 0.20 0.39 <0.0001 GOSE at 6 mo - GR/MD - SD/V/D - Missing/lost to f/u - 30 (48%) - 30 (48%) - 3 (4%) - 4 (12%) - 26 (76%) - 4 (12%) 0.001 Mortality at 6 mo - Survived - Died - Missing/lost to f/u - 57 (90%) - 3 (4%) - 3 (4%) - 21 (62%) - 9 (26%) - 4 (12%) 0.001 Bold denotes p≤0.05; italic denotes p≤0.1. GCS: Glasgow Coma Scale. GOSE GR/MD/SD/V/D: Good Recovery/Moderate Disability/Severe Disability/Vegetative State/Dead. IQR: interquartile range. IMPACT: International Mission for Prognosis and Analysis of Clinical Trials in TBI. Table 1B: Secondary injury measures for patients in the two NfL trajectory groups Serum NfL TRAJ GROUP Low NfL High NfL p-value Number of patients 63 34 Average ICP (mean, mmHg) 14.75 18.65 0.01 ICP - Never >25 - > 25mmHg at least once - 16 (25%) - 47 (75%) - 10 (29%) - 24 (71%) 0.81 ICP duration > 25 mmHg (h) 9.66 20.94 0.01 ICP - Never > 30 - > 30 mmHg at least once - 25 ( 40%) - 38 (60%) - 15 (44%) - 19 (56%) 0.14 ICP duration > 30 mmHg (h) 2.49 9.94 0.0069 Required mannitol: - No - Yes - 24 (38%) - 39 (62%) - 11 (33%) - 23 (67%) 0.66 Required barbiturate coma: - No - Yes - 57 (91%) - 6 (9%) - 24 (71%) - 10 (29%) 0.01 Required decompressive craniectomy - No - Yes - 46 (73%) - 17 (27%) - 17 (50%) - 17 (50%) 0.02 Average MAP (mmHg) 90.65 91.38 0.60 MAP: - Never < 70 mmHg - < 70 mmHg at least once - 26 (41%) - 37 (59%) - 8 (24%) - 26 (76%) 0.11 MAP duration < 70 mmHg (h) 3.4 4.7 0.34 Average CPP (mmHg) 75.58 72.36 0.06 CPP: - Never <50mmHg - < 50mmHg at least once - 23 (37%) - 40 (63%) - 9 (26%) - 25 (74%) 0.37 CPP duration <50mmHg (h) 2.88 7.58 0.01 Average PbtO 2 (mmHg) 33.06 26.5 0.05 PbtO 2 : - Never <10 mmHg - <10mmHg at least once - 38 (62%) - 23 (38%) - 18 (55%) - 15 (45%) 0.5 PbtO 2 duration <10mmHg (mean, hours) 4.75 (missing 2) 14.6 (missing 1) 0.08 Average Sjv02 76.93 Missing 14 74.83 Missing 6 0.20 SjvO 2 - Never <50% - <50% at least once - 38 (78%) - 11 (22%) - 17 (61%) - 11 (39%) 0.12 SjvO 2 duration <50% (h) 0.75 1.17 0.23 Delayed hematoma - Absent - Present - 44 (70%) - 19 (30%) - 22 (65%) - 12 (35%) 0.65 Worst Marshall CT - Diffuse injury 2 - Diffuse injury 3 - Diffuse injury 4 - Evacuated Mass Lesion - Unevacuated Mass Lesion - 28 - 14 - 1 - 17 - 3 - 7 - 13 - 1 - 11 - 2 0.0014 Bold denotes p≤0.05; italic denotes p≤0.1.ICP: intracranial pressure. MAP: Mean arterial pressure. CPP: Cerebral perfusion pressure. PbtO2: Brain tissue oxygen partial pressure. SjvO2: Jugular venous oxygen saturation. CT: Computed Tomography Table 2. GOSE outcome models of (A) serum NfL means by time epoch, and (B) both serum NfL mean and GOSE specific ICU injury burden score at all time epochs . (A) NfL Mean → 6-month GOSE outcome (individual regressions) OR CI p-value AUC Serum NfL Mean (day 0) 1.007 1.001-1.014 0.0326 0.646 Serum NfL Mean (days 1-5) 1.005 1.001-1.008 0.0108 0.660 Serum NfL Mean (days 6-10) 1.002 1.00-1.004 0.0111 0.664 (B) NfL Mean (Adjusted for ICU injury burden score) → 6-month GOSE outcome OR CI p-value AUC Serum NfL (day 0) 1.004 0.99-1.01 0.2497 0.779 ICU injury burden (day 0) 3.418 1.72-6.79 0.0004 OR CI p-value AUC Serum NfL (days 1-5) 1.004 1.0-1.07 0.0362 0.733 ICU injury burden (days 1-5) 3.814 1.35-10.75 0.0013 OR CI p-value AUC Serum NfL (days 6-10) 1.002 1.0-1.004 0.0223 0.770 ICU injury burden (days 6-10) 2.97 1.25-7.07 0.0141 Each row represents and individual univariate binary logistic regression in A) with the corresponding odds ratio (OR), 95% confidence interval (CI), p-value, and area under the curve (AUC). Each pair of NfL and ICU injury burden by time epoch represent an individual multivariate binary logistic regression in B) with the corresponding OR, CI, p-value, and AUCs. Model results odds of an unfavorable GOSE=1-4. Bold denotes p≤0.05. Table 3. Individual regression models to A) 6-month GOSE outcome, B) Day 0-10 NfL TRAJ group, and C-F) ICU injury burden at all time epochs A) 6-month GOSE (dichotomized) B) Day 0-10 NfL TRAJ C) Day 0 ICU injury burden D) Day 1-5 ICU injury burden E) Day 6-10 ICU injury burden F) Day 0-10 ICU injury burden OR [CI] p-value OR [CI] p-value B (SE) p-value B (SE) p-value B (SE) p-value B (SE) p-value Age 1.034 [1.0-1.07] p=0.0382 1.012 [0.98-1.04] p=0.4316 0.01 (0.009) p=0.0889 0.0038 (0.004) p=0.2896 0.0001 (0.004) p=0.9860 0.001 (0.005) p=0.8994 Sex 0.826 [0.29-2.34] p=0.7179 1.098 [0.37-3.24] p=0.8656 -0.54 (0.35) p=0.1259 -0.088 (0.13) p=0.4979 -0.15 (0.05) p=0.3212 -0.19 (0.19) p=0.3334 ER GCS Sum 0.74 [0.63-0.87] p=0.0003 0.785 [0.663-0.93] p=0.0049 -0.09 (0.05) p=0.0454 -0.008 (0.02) p=0.6365 -0.03 (0.02) p=0.1274 -0.04 (0.03) p=0.1681 GCS (best in 24h) 0.725 [0.58-0.9] p=0.0037 0.779 [0.62-0.98] p=0.0357 -0.05 (0.06) p=0.4336 -0.01 (0.02) p=0.6485 -0.01 (0.03) p=0.7074 -0.03 (0.04) p=0.4540 CT burden 3.73 [1.99-6.98] p=<0.0001 2.24 [1.19-4.24] p=0.0130 0.72 (0.16) p<0.0001 0.34 (0.06) p<0.0001 0.42 (0.06) p<0.0001 0.56 (0.08) p<0.0001 Day 0 ICU injury burden factors: Duration (h) CPP <50, Duration (h) PbtO2 25, mannitol dose required, and decompressive craniectomy and/or barbiturate coma dose required Day 1-5 ICU injury burden factor: duration (h) PbtO2 <10, decompressive craniectomy and/or barbiturate coma dose required. Day 6-10 ICU injury burden factors: mannitol dose required, decompressive craniectomy and/or barbiturate coma dose required Day 0-10 ICU injury burden factors: duration (h) CPP<50, Duration (h) PbtO2<10, and decompressive craniectomy and/or barbiturate coma dose required. Bold denotes p≤0.05; italic denotes p≤0.1. GCS: Glasgow Coma Scale. CT: Computed Tomography. ICU: Intensive Care Unit. OR: Odds Ratio. CI: Confidence interval. SE: Standard Error Table 4. Covariate adjusted logistic regression models of dichotomized 6-month GOSE outcome. 6-month GOSE (dichotomized) Baseline Characteristic Model (AUC=0.787) NfL Risk Model (AUC=0.803) NfL Risk + ICU injury burden Model (AUC=0.819) Independent variables OR CI p-value OR CI p-value OR CI p-value Age 1.01 0.97-1.05 0.5852 1.008 0.97-1.05 0.6828 1.08 0.97-1.06 0.4267 Gender (Male vs. Female) 1.38 0.39-4.87 0.6217 1.303 0.36-4.68 0.6853 1.10 0.28-4.33 0.8921 GCS, best in 24h 0.74 0.58-0.93 0.0104 0.74 0.57-0.96 0.0241 0.69 0.58-0.92 0.0106 CT burden score 3.60 1.76-7.35 0.0004 3.02 1.42-6.42 0.0040 2.04 0.87-4.78 0.1007 NfL risk TRAJ - - - 2.57 0.89-7.42 0.0823 1.95 0.65-5.92 0.2361 Overall Day 0-10 ICU injury burden score (GOSE specific) - - - - - - 2.24 1.02-4.92 0.0447 Bold denotes p≤0.05. Italic denotes p≤0.1. GCS: Glasgow Coma Scale. CT: Computed Tomography. ICU: Intensive Care Unit. OR: Odds Ratio. CI: Confidence interval. Supplementary Files STROBEchecklistcohortNfLmanuscript.docx Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 30 Oct, 2025 Reviewers invited by journal 30 Oct, 2025 Editor invited by journal 29 Oct, 2025 Editor assigned by journal 27 Oct, 2025 First submitted to journal 24 Oct, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7909088","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":537685223,"identity":"f8a47463-fda3-47b4-9d5a-15c60b5ebfc1","order_by":0,"name":"Spyridoula Tsetsou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABE0lEQVRIiWNgGAWjYLCCBDYGBj4El/kYhD5AQAsQMTYwMBgAuWxphLUwoGrhMcOrRX52j9mHB2U2iW3sB9gf/qj4Y88vkfPtwc8cBjm+GwlYtRjcOWM8I+FcWmIbTwJjM88Zg8SZM3K3G/ZuYzCWxKVFIseYIbHtcGIbA1ALY5tBgsGN3G3SjNsYEjfg0CI/A6aF/wFj489/BvYGN3KegbTU49LCcAOmRSKBsYG3wYBxw40cNpAWoHW4/HKsmAHoF+M2iYeNs3mOGSfO7HlmDvSLhOHMMw9whFjzZsYfZTay/fzJBz7+qJGz52dPfvbg5zYbeb7jOBwmwWEAZYGiBUkcu3KwFDt260fBKBgFo2AUwAEAS/lhimISlqIAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0003-4521-5745","institution":"Baylor College of Medicine","correspondingAuthor":true,"prefix":"","firstName":"Spyridoula","middleName":"","lastName":"Tsetsou","suffix":""},{"id":537685224,"identity":"40b2138b-17b8-48f5-a74c-82250898ad1e","order_by":1,"name":"Leah E McQuillan","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Leah","middleName":"E","lastName":"McQuillan","suffix":""},{"id":537685225,"identity":"9e8afc90-2188-4e08-bd0d-8f1790842bb7","order_by":2,"name":"David J Barton","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"David","middleName":"J","lastName":"Barton","suffix":""},{"id":537685226,"identity":"2c6f3d7c-7279-442c-abde-7ce56c1d0b6f","order_by":3,"name":"John Williamson","email":"","orcid":"","institution":"University of Florida","correspondingAuthor":false,"prefix":"","firstName":"John","middleName":"","lastName":"Williamson","suffix":""},{"id":537685227,"identity":"c5726b3c-ec46-4cb2-bb15-023403240852","order_by":4,"name":"Damon G Lamb","email":"","orcid":"","institution":"University of Florida","correspondingAuthor":false,"prefix":"","firstName":"Damon","middleName":"G","lastName":"Lamb","suffix":""},{"id":537685228,"identity":"3ca6295d-60d6-4611-a242-8ed540dd716a","order_by":5,"name":"Firas Kobeissy","email":"","orcid":"","institution":"Morehouse School of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Firas","middleName":"","lastName":"Kobeissy","suffix":""},{"id":537685229,"identity":"cfee5049-a201-4fc1-badc-9c57d9dca306","order_by":6,"name":"Guangzheng Cai","email":"","orcid":"","institution":"Morehouse School of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Guangzheng","middleName":"","lastName":"Cai","suffix":""},{"id":537685230,"identity":"dc3f2672-1d46-4b04-81aa-5b085d7ab78e","order_by":7,"name":"Richard Rubenstein","email":"","orcid":"","institution":"SUNY Downstate: SUNY Downstate Health Sciences University","correspondingAuthor":false,"prefix":"","firstName":"Richard","middleName":"","lastName":"Rubenstein","suffix":""},{"id":537685231,"identity":"c5f13685-1a70-4ca0-af2d-c3e207bd8a9e","order_by":8,"name":"Kevin K.W. Wang","email":"","orcid":"","institution":"Morehouse School of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Kevin","middleName":"K.W.","lastName":"Wang","suffix":""},{"id":537685232,"identity":"d3733345-ae6c-4dce-9f8f-67bc50643062","order_by":9,"name":"Claudia S. Robertson","email":"","orcid":"https://orcid.org/0000-0002-8509-678X","institution":"Baylor College of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Claudia","middleName":"S.","lastName":"Robertson","suffix":""},{"id":537685233,"identity":"c74794c3-fa49-4fb4-bbc3-bb10a7b75ba6","order_by":10,"name":"Amy K Wagner","email":"","orcid":"","institution":"University of Pittsburgh","correspondingAuthor":false,"prefix":"","firstName":"Amy","middleName":"K","lastName":"Wagner","suffix":""}],"badges":[],"createdAt":"2025-10-20 23:08:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7909088/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7909088/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":95805797,"identity":"c65acf60-3195-4fd1-a350-4f56358ffc92","added_by":"auto","created_at":"2025-11-13 08:46:51","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":22041,"visible":true,"origin":"","legend":"","description":"","filename":"Table1NCC.docx","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/9e6c443aad2a5b3e52efb241.docx"},{"id":95805878,"identity":"008feeee-e875-4358-b60e-9111c1ef9935","added_by":"auto","created_at":"2025-11-13 08:47:03","extension":"docx","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":17506,"visible":true,"origin":"","legend":"","description":"","filename":"Table2NCC.docx","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/84c5c9d18dd09a441e206cff.docx"},{"id":95806066,"identity":"39df2f5f-0b03-4714-87f0-eb5002b9d969","added_by":"auto","created_at":"2025-11-13 08:47:14","extension":"docx","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":16457,"visible":true,"origin":"","legend":"","description":"","filename":"Table3NCC.docx","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/24ec2af6f33154ccc0b51c63.docx"},{"id":95806253,"identity":"846d013d-4ce6-48de-8c94-97f7b43a494a","added_by":"auto","created_at":"2025-11-13 08:47:21","extension":"docx","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":17610,"visible":true,"origin":"","legend":"","description":"","filename":"Table4NCC.docx","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/5bc45904e69382c33c005c93.docx"},{"id":95805885,"identity":"5226b3c3-be02-4e4f-a288-e576d1963725","added_by":"auto","created_at":"2025-11-13 08:47:03","extension":"xml","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":15954,"visible":true,"origin":"","legend":"","description":"","filename":"necaNECAD2501146.xml","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/8de20d217866e77f418036aa.xml"},{"id":95806284,"identity":"9548cbac-63db-4ebb-8a6d-ac68065c891e","added_by":"auto","created_at":"2025-11-13 08:47:22","extension":"xml","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1359,"visible":true,"origin":"","legend":"","description":"","filename":"NECAD250114617898.go.xml","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/14efc7b716e6b77c82a5aa1c.xml"},{"id":95805928,"identity":"3ec80263-adde-494c-92bc-98927c2cb7ca","added_by":"auto","created_at":"2025-11-13 08:47:07","extension":"xml","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":922,"visible":true,"origin":"","legend":"","description":"","filename":"NECAD2501146Import.xml","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/0b0b0a95e881ad42b111f31b.xml"},{"id":95805988,"identity":"b7faaca4-2419-4a49-95bd-8ebbb9f49fd6","added_by":"auto","created_at":"2025-11-13 08:47:11","extension":"xml","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":170417,"visible":true,"origin":"","legend":"","description":"","filename":"NECAD25011460enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/5f8b654241a71bd7d3f045dd.xml"},{"id":95805967,"identity":"dda51bee-fae1-4468-b860-54e4fc0c25e4","added_by":"auto","created_at":"2025-11-13 08:47:10","extension":"pdf","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":184240,"visible":true,"origin":"","legend":"","description":"","filename":"Figure1Nfl.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/6d9a4cfa721ef706b86b2465.pdf"},{"id":95806028,"identity":"c8888033-c325-4aba-aaf0-edda511bd823","added_by":"auto","created_at":"2025-11-13 08:47:12","extension":"pdf","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":477748,"visible":true,"origin":"","legend":"","description":"","filename":"Figure2NfL.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/a419c0e4a264f3561e0219b5.pdf"},{"id":95805825,"identity":"8491c2d8-38bf-4fcd-ba92-36082f47f29b","added_by":"auto","created_at":"2025-11-13 08:46:57","extension":"pdf","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":267675,"visible":true,"origin":"","legend":"","description":"","filename":"Figure3NfL.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/3157921462714602424ac65f.pdf"},{"id":95805792,"identity":"da5961f3-2d1b-495a-8c3d-ca848ded47f1","added_by":"auto","created_at":"2025-11-13 08:46:50","extension":"jpeg","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":10444,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/73ec2181df2c57c9bb75f4ee.jpeg"},{"id":95805808,"identity":"68661d45-c1bc-499c-8c46-2734cd748d3a","added_by":"auto","created_at":"2025-11-13 08:46:55","extension":"png","order_by":18,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":7915,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/fc0bafecc5d3e8b99f3e3842.png"},{"id":95805964,"identity":"f8839f8b-43cf-4873-a0a7-9bc9be7f929e","added_by":"auto","created_at":"2025-11-13 08:47:10","extension":"xml","order_by":19,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":168474,"visible":true,"origin":"","legend":"","description":"","filename":"NECAD25011460structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/f33684e99602380897dc2bd0.xml"},{"id":95806152,"identity":"c4986a4b-9da5-4f07-8931-9441fb8e8875","added_by":"auto","created_at":"2025-11-13 08:47:17","extension":"html","order_by":20,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":183884,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/9b7927b7b92159e92a4b61df.html"},{"id":95806146,"identity":"fc68c7d3-2ba8-466c-a6b5-bf6cd6650638","added_by":"auto","created_at":"2025-11-13 08:47:17","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":191348,"visible":true,"origin":"","legend":"\u003cp\u003eA) Consort diagram of patients included for analysis. B) Serum NfL trajectory\u003c/p\u003e\n\u003cp\u003egroups. Two subgroups with high (n=34) vs low (n=63) NfL levels over time. Both groups\u003c/p\u003e\n\u003cp\u003eexhibited increasing levels over the 10-day monitoring period. Data shown are mean ± standard\u003c/p\u003e\n\u003cp\u003edeviation\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/c54685cd3f1e93115d365fd5.jpg"},{"id":95805897,"identity":"6e8ff12b-9a47-46d6-a915-6d17186879c7","added_by":"auto","created_at":"2025-11-13 08:47:04","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":791539,"visible":true,"origin":"","legend":"\u003cp\u003eA) Cross-Leg regression schematic using standardized biomarker means and 6-\u003c/p\u003e\n\u003cp\u003emonth GOSE anchored ICU injury burden score. Each arrow represents an unadjusted linear\u003c/p\u003e\n\u003cp\u003eregression with the corresponding beta-value and p-value. Bolded p-values signify significance\u003c/p\u003e\n\u003cp\u003elower than p \u0026lt;0.05 threshold. Factors included in each ICU injury burden anchored to GOSE, by\u003c/p\u003e\n\u003cp\u003etime epoch are listed below:\u003c/p\u003e\n\u003cp\u003e•Day 0: Duration (h) CPP \u0026lt;50, Duration (h) PbtO2 \u0026lt;10, Duration (h) ICP \u0026gt;25, mannitol\u003c/p\u003e\n\u003cp\u003edose required, and decompressive craniectomy and/or barbiturate coma dose required.\u003c/p\u003e\n\u003cp\u003e•Day 1-5: Duration (h) PbtO2 \u0026lt;10, and decompressive craniectomy and/or barbiturate coma\u003c/p\u003e\n\u003cp\u003edose required.\u003c/p\u003e\n\u003cp\u003e•Day 6-10: Mannitol dose required, and decompressive craniectomy and/or barbiturate\u003c/p\u003e\n\u003cp\u003ecoma dose required.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/8afdacba9553f40eac7ded82.jpg"},{"id":95805847,"identity":"d1928977-dad2-4ffc-8760-6c35604aadc3","added_by":"auto","created_at":"2025-11-13 08:46:59","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":512561,"visible":true,"origin":"","legend":"\u003cp\u003eA) Mediation model of Day 0 serum NfL mean and Day 1-5 ICU injury burden, B)\u003c/p\u003e\n\u003cp\u003eDay 1-5 serum NfL mean and Day 6-10 ICU injury burden, and C) Serum Day 0-10 NfL\u003c/p\u003e\n\u003cp\u003eTRAJ and Day 0-10 ICU injury burden to dichotomized 6-month unfavorable GOSE (GOSE=1-4 vs. GOSE=5-8). Covariates included age and sex for all individual models. B(SE) or\u003c/p\u003e\n\u003cp\u003eOR[CI] and p-values are shown for each model. The model labeled direct includes ICU injury\u003c/p\u003e\n\u003cp\u003eburden in the model to test for the mediation effect. Mediation percentage is shown. P\u0026lt;0.05 are\u003c/p\u003e\n\u003cp\u003ebolded.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/07d52568d8da066ac2c83d04.jpg"},{"id":95818800,"identity":"4f76ed42-b7ab-4de1-aed0-952aeebccf37","added_by":"auto","created_at":"2025-11-13 10:33:33","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3569563,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/03cb6d0e-7871-4178-b653-fe7c8ed4a346.pdf"},{"id":95805865,"identity":"0f27b291-8e21-4af8-9664-82a1398203ae","added_by":"auto","created_at":"2025-11-13 08:47:01","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":31872,"visible":true,"origin":"","legend":"","description":"","filename":"STROBEchecklistcohortNfLmanuscript.docx","url":"https://assets-eu.researchsquare.com/files/rs-7909088/v1/4b5286b35f1b356344c33c5e.docx"}],"financialInterests":"","formattedTitle":"Neurofilament Light Protein: a marker for injury severity, clinical course and outcome following moderate-severe Traumatic Brain Injury","fulltext":[{"header":"Introduction","content":"\u003cp\u003eTraumatic brain injury (TBI) remains a leading cause of death and major disability, with increasing incidence and health-care costs\u003csup\u003e\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. The discovery of blood-based biomarkers, as potential prognostic, monitoring and diagnostic tools, has led to increasing interest and research in this field\u003csup\u003e\u003cspan additionalcitationids=\"CR5 CR6\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase L1 protein (UCH-L1) levels in blood during the acute phase of TBI, have been approved as an aid in the decision to obtain Computed Tomography (CT) imaging among individuals clinically assessed as having mild TBI\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. The neurotrauma field has had a rising interest in exploring the utility of neurofilament light (NfL), a marker showing high sensitivity for axonal injury that maps to subacute/chronic brain injury and MRI brain atrophy after moderate/severe TBI (msTBI) \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. However, less is known about the utility this marker may have in reflecting ongoing secondary brain injury among critically ill in the Intensive Care Unit (ICU) with msTBI.\u003c/p\u003e\u003cp\u003eNeurofilament (Nf) proteins are neuron-specific intermediate filaments abundant in axons. During axonal injury, they dissociate from the cytoskeleton, cause cellular membrane integrity loss and accumulate in extracellular fluid, cerebrospinal fluid (CSF) and blood. They are highly specific to axonal injury\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. NfL, Nf-heavy (NfH) and neurofilament-medium form the three subunits of Nf proteins. NfL is considered to be a biomarker reflecting neurodegeneration and is associated with cognitive impairment in patients with mild cognitive impairment and dementia (Alzheimer\u0026rsquo;s disease, frontotemporal dementia)\u003csup\u003e\u003cspan additionalcitationids=\"CR15 CR16\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. NfL remains high in CSF and serum following TBI\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e,\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e and is associated with progressive neurodegeneration in this population\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. Prior work\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e involving patients with msTBI explored the potential for Nf proteins to predict 6-months cognitive outcome in this patient group. Interestingly, the NfL levels were measured later post-injury (16\u0026ndash;90 days post injury) did not correlate with the early NfL levels (0\u0026ndash;15 days post injury) or 6-months cognitive performance\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eHowever, NfL has been linked with 6- or 12-months functional outcome, in all severities of TBI\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. Unlike other biomarkers, blood NfL levels continue to rise up to 6 weeks post msTBI\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e, with the peak NfL level linked to poor 6-months functional outcome and to brain atrophy on MRI\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. In related work\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e, we recently conducted NfL temporal kinetic profile characterizations from the time of injury through subacute and chronic recovery to find that serum NfL level peak\u0026thinsp;~\u0026thinsp;30 days post-injury; also, both acute and chronic serum NfL levels are associated with global recovery at 6- and 12-months post-injury.\u003c/p\u003e\u003cp\u003eWe previously screened multiple serum biomarkers, for associations with clinical physiological variables measured over time in the acute injury phase\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. This work highlighted that GFAP levels in the first 10 days were associated with intracranial pressure (ICP) and cerebral perfusion pressure (CPP) summary variables, injury type presentation on CT scans, mortality and recovery, and demonstrated rapidly declining GFAP levels after initial peak correlated with secondary insults. Also, averaged NfL serum levels in the first 10 days were associated with acute physiological parameters, specifically with brain hypoxia (longer duration of PbtO2\u0026thinsp;\u0026lt;\u0026thinsp;10 mmHg) and ICP treatment\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. However, the role of NfL within the ICU clinical course remains poorly understood. Evidence supporting its utility in ICU-based clinical decision-making is limited, and the potential causal relationship between ICU injury burden profiles and outcomes has not been explored.\u003c/p\u003e\u003cp\u003eThus, we evaluated how acute phase, serial NfL levels correlate with the ICU clinical course and outcomes in patients with msTBI. We hypothesized that NfL levels reflect axonal injury sensitive to the extent of secondary brain injury, as indicated by ICU-associated clinical data, and 6-months global outcomes.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStudy design and participants\u003c/h2\u003e\u003cp\u003eWe used archived blood samples that were originally collected as part of our clinical trial of erythropoietin (clinicaltrials.gov NCT00313716). Inclusion criteria for this study were: 1) age at least 15 years old, 2) motor GCS \u0026le; 5 after resuscitation related to a closed TBI and 3) ability to enroll within 6 hours from trauma. Exclusion criteria included 1) patients with GCS of 3 with fixed and dilated pupils, 2) penetrating trauma, 3) pregnancy (positive urine test) and 4) life-threatening injuries or severe pre-existing comorbidities. Patients enrolled in the trial were randomly assigned to treatment with erythropoietin or placebo and to a transfusion threshold of 7 or 10 g/dL using a 2x2 factorial design. Blood samples were collected every 6 hours for the first 24 hours after injury and then once daily through day 10 post-injury. Samples were centrifuged for 15 minutes at 1500g, and serum was removed and stored at -80\u003csup\u003eo\u003c/sup\u003e C until analyzed. Patients were offered the option to have research blood samples banked for future studies analyses after clinical trial analysis were completion. Of the 200 trial participants, 114 opted to allow future use of the samples and their associated clinical data. Samples were available on 103 patients but only 97 had at least two sample biomarker measurements available during the biosample collection period, and included in this analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eBaylor College of Medicine Institutional Review Board approved waived consent for the use of these archived, de-identified blood samples (protocol H-44131).\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eBiomarker assays\u003c/h3\u003e\n\u003cp\u003eAs described in prior work\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e, we assayed NfL with an ultrasensitive immunoassay using digital array technology (Single Molecule Arrays SiMoA)-based Human Neurology 4-Plex B assay (N4PB; Item 103345). All SiMoA assays were run on the SR-X benchtop assay platform (Quanterix Corp., Lexington MA) at the University of Florida (Gainesville, FL) according to manufacturer\u0026rsquo;s instructions. The lower limit of quantification (LLOQ), lower limit of detection (LLOD), and dynamic range were 0.625 pg/mL, 0.097 pg/mL, and 0.0971-10,000 pg/mL for NfL, respectively. Inter-assay and intra-assay % CV were 4.6\u0026ndash;4.9% and 3.5\u0026ndash;7.5%, respectively for NfL.\u003c/p\u003e\n\u003ch3\u003eOutcome measures and other ICU variables\u003c/h3\u003e\n\u003cp\u003eDemographics and injury severity variables included age, sex, race, GCS sum and motor component (pre- and post-resuscitation), injury severity scale (ISS), clinical exam including pupillary reactivity, prehospital hypotension and hypoxia, and acute CT findings (initial and worst Marshall CT category and a CT burden score). International Mission for Prognosis and Analysis of Clinical Trials in TBI (IMPACT) risk of poor outcome and mortality were also calculated\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. Glasgow Outcome Scale-Extended (GOSE) was used to determine 6-months outcome. For these analyses, GOSE was dichotomized as favorable (score 5\u0026ndash;8) and unfavorable outcome (score 1\u0026ndash;4).\u003c/p\u003e\u003cp\u003eCT reports for the admission scan were reviewed by neurointensivist and/or neurosurgeon attendings and categorized for pre-specified brain lesion types: intraventricular hemorrhage (IVH), intraparenchymal hemorrhage (IPH), subdural hematoma (SDH), subarachnoid hemorrhage (SAH), epidural hematoma (EDH) or diffuse brain injury (DBI), contusion and midline shift.\u003c/p\u003e\u003cp\u003eDetails about the acute care, clinical events and treatment were extracted from the electronic medical record (EMR), prepared, and summarized for data analysis. Data including ICP, mean arterial pressure (MAP), CPP, jugular venous oxygen saturation (SjvO2), and brain tissue pO2 (PbtO2) were collected and analyzed. The physiological variables were recorded hourly for as long as the ICP was being monitored over the first 10 days of injury. We used average ICP, duration of time that ICP was higher than 25 mmHg and also higher than 30 mmHg, duration of time of CPP was lower than 50 mmHg, duration of time that PbtO2 was less than 10 mmHg, and duration of time that SjvO2 less than 50%. Average values of other parameters and duration of time of abnormal values were collected as well. Treatments to control ICP (e.g. mannitol, barbiturate coma, decompressive craniectomy) were also collected and analyzed. Sustained increases in ICP were treated according to local protocols and the three-tiered algorithm, including but not limited to sedative medications, CSF drainage, hyperventilation, hyperosmotic therapy, barbiturate induced coma, surgical decompression. Physiological variables data and clinical events were binned into 24-h increments to examine changes over time.\u003c/p\u003e\n\u003ch3\u003eICU injury burden score\u003c/h3\u003e\n\u003cp\u003eThe ICU injury burden score was calculated as described in our prior work\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. To calculate the ICU injury burden score, the physiological data were also binned into 24-h increments, and the following conditions known to be associated with poor outcome were quantified: duration of time that ICP\u0026thinsp;\u0026gt;\u0026thinsp;25 mmHg, CPP\u0026thinsp;\u0026lt;\u0026thinsp;50 mmHg, and PbtO2\u0026thinsp;\u0026lt;\u0026thinsp;10 mmHg; mannitol dose administration, implementation of barbiturate coma and/or decompressive craniectomy. Frequency of out-of-range values was evaluated over the full 10-day time course post-injury and in three separate time epochs: the first 24 h, days 1\u0026ndash;5, and days 6\u0026ndash;10.\u003c/p\u003e\u003cp\u003eGiven the inter-related nature of acute care physiology data, and inherent collinearity associated with these multiple ICU factors included in the modeling approach, we generated a composite score reflecting cumulative ICU related physiological injury burden rather than assessing individual factors with respect to biomarker levels and long-term outcome. Generating an aggregate ICU injury burden score through data reduction techniques reduces noise and provides a cumulative measure of relevant injury factors that reflect ICU practice patterns over time. A series of simple logistic regressions were conducted considering ICU injury factors over three-time epochs (first 24h-day 0, days 1\u0026ndash;5, and days 6\u0026ndash;10) with respect to 6-months GOSE and 6-months mortality.\u003c/p\u003e\u003cp\u003eICU factors associated with outcome with a threshold p-value of \u0026lt;\u0026thinsp;0.1 were identified using unadjusted linear regression and included in the respective ICU injury burden score derivation. A ridge regression that included each significant ICU factor stated above was then applied to create the ICU injury burden scores for each respective time epoch. This method yielded β-weight estimates for each factor in the model, with smaller β-weights reflecting smaller effect sizes. Appropriate to our research question and analytical approach, this method also penalizes multicollinearity when present, which leads to more reliable β-weight estimates and reduced standard error (SE). This penalty term reduces the impact of multicollinearity by shrinking correlated predictors\u0026rsquo; coefficients toward zero in a balanced way, thereby producing more stable and interpretable estimates. The most stable and frequent ridge penalty parameter was selected after iterating a sixfold cross validation 1000 times using the R package glmnet\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. The coefficients generated were then multiplied to each individual\u0026rsquo;s set of relevant ICU injury factors and summed to produce the ICU injury burden score in each time epoch.\u003c/p\u003e\n\u003ch3\u003eCT burden score\u003c/h3\u003e\n\u003cp\u003eAn admission CT injury burden score was also generated using ridge regression as described previously\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. CT reports for the admission scan were reviewed and categorized as mentioned above. The injury types associated with GOSE in univariate logistic regression (midline shift, SAH, SDH, EDH, IPH, IVH, DBI and contusion) were selected for inclusion in the weighted sum reflecting CT injury burden.\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eGroup-based trajectory analysis\u003c/h2\u003e\u003cp\u003eWe previously reported temporal profiles for days 0\u0026ndash;10 NfL\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e using group-based trajectory (TRAJ) analysis\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e for subjects with \u0026ge;\u0026thinsp;2 sample timepoints in the first 10 days post-injury. Polynomial orders were assigned to group data from each cohort based on minimizing Bayesian Information Criterion (BIC) and ensuring posterior probability\u0026thinsp;\u0026gt;\u0026thinsp;80% using CNORM distribution (SAS software, Proc TRAJ). These TRAJ group memberships were leveraged in this study for further analysis of relationships to ICU injury burden score, demographic and clinical factors, and global outcome (GOSE).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eDescriptive analyses for continuous variables used mean, standard error (SE) of the mean, median, and interquartile range (IQR). Frequencies and percentages were used to quantify categorical variables. Spearman rank correlation tests were applied to compare continuous variables that were not normally distributed. Univariate and multivariate linear and binary logistic regression models were used, where applicable, to assess associations between NfL means, NfL TRAJ group membership, ICU injury burden score, and demographic and clinical covariates with 6-months outcomes (GOSE). ICU injury burden score was evaluated, using the Baron and Kenny approach\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e for a possible mediating relationship between NfL mean levels\u0026mdash;and also TRAJ group membership\u0026mdash;and 6-month outcome. The area under the receiver operating characteristic curve (AUC) was used to evaluate the discriminative performance of each model. DeLong\u0026rsquo;s test was employed to compare the AUCs between models, assessing whether differences in model performance were significant. Multivariate models assessing ICU injury burden as a mediator were adjusted for age and sex. Results were reported when the total effect was greater than the direct effect and a p-value threshold less than 0.1 was met. Mediation percentage was calculated as the proportion of the indirect effect to the total effect, shown below. Statistical analyses were conducted using SAS Version 9.4 (Cary, North Carolina), R and GraphPad Prism Version 10 (Boston, Massachusetts). For all analyses, a p-value\u0026thinsp;\u0026le;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"601\" height=\"97\"\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eDemographics, clinical, ICU variables and outcomes by NfL TRAJ\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNinety-seven (97) patients had samples and data available and were used for analysis. TRAJ analysis identified two distinct subgroups with \u003cem\u003ehigh\u003c/em\u003e (n=34) vs \u003cem\u003elow\u003c/em\u003e (n=63) NfL levels; both groups exhibited increasing levels over the 10-day monitoring period \u003cstrong\u003e\u003cem\u003e(Figure 1B\u003c/em\u003e\u003c/strong\u003e). \u003cstrong\u003e\u003cem\u003eTable 1A\u003c/em\u003e\u003c/strong\u003e shows the demographics, clinical and radiological characteristics of the two groups. The \u003cem\u003ehigh\u003c/em\u003e TRAJ group had worse admission GCS (p=0.0009), higher CT burden scores (p=0.0086), more frequently compressed basal cisterns on admission brain CT (p=0.03), and prehospital hypotension (p=0.046). \u0026nbsp;Higher IMPACT score for risk of poor outcome and risk of mortality was also observed for those in high TRAJ group (p\u0026lt;0.0001, p \u0026lt;0.0001 respectively). Similarly, \u003cem\u003ehigh\u003c/em\u003e TRAJ group membership was associated with an increased mortality rate (p=0.001) and worse clinical outcome with GOSE assessment at 6 months (p=0.001).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eTable 1B\u003c/em\u003e\u003c/strong\u003e shows the ICU physiological variables for patients of the two TRAJ groups. Average ICP was higher for in the \u003cem\u003ehigh\u003c/em\u003e TRAJ group (p=0.01), and this group had also longer duration of time of ICP \u0026gt;25mmHg (p=0.01) and \u0026gt;30mmHg (p=0.0069). \u003cem\u003eHigh\u003c/em\u003e TRAJ group members also was associated with more prolonged cerebral hypoperfusion (7.58hrs vs 2.88hrs, p=0.01), though with only trend-level differences observed in the average CPP value. Patients in the \u003cem\u003ehigh\u003c/em\u003e TRAJ group more frequently required barbiturate coma (p=0.01) or decompressive surgery (p=0.02) for controlling ICP.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCross-sectional NfL and ICU injury burden relationships\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDay 0, Day 1-5, Day 6-10, and Day 0-10 means for CSF and serum NfL were all screened against their respective ICU injury burden by time epoch (not shown). Serum NfL, but not CSF, levels were associated with ICU injury burden at all time points.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eFigure 2A\u003c/em\u003e\u003c/strong\u003epresents a cross-leg framework of univariate linear regression models that show the consistent interrelatedness between serum NfL and ICU injury burden factors over the first 10 days post-injury. Day 0 NfL means were significantly associated with Day 1-5 NfL means, and subsequently Day 1-5 NfL means to Day 6-10 NfL means (p\u0026lt;0.001 both comparisons). ICU injury burden demonstrated a similar relationship between adjacent time epochs (p\u0026lt;0.001 both comparisons). Additionally, mean NfL and ICU injury burden scores were significantly related to each other within time epoch (Day 0 NfL to Day 0 ICU injury burden) and across time epochs (day 0 to Day 1-5, and Day 1-5 to Day 6-10).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNfL, ICU injury burden, demographic and clinical factors relationship\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA correlational heatmap (\u003cstrong\u003e\u003cem\u003eFigure 2B\u003c/em\u003e\u003c/strong\u003e) presents pairwise comparisons (by Spearman rank correlation) between NfL means and ICU injury burden scores at all time epochs (Days: 0, 1-5, 6-10, 0-10) with GCS, age and CT burden. Each cell shows the correlation coefficient (r) for the pairwise comparison. Mean NfL levels were positively associated with ICU injury burden and CT burden, and negatively associated with GCS. ICU injury burden was also negatively associated with GCS and positively associated with CT burden at all time epochs. Age had a stronger relationship to ICU injury burden at Day 0 and 1-5 than later timepoints. NfL means and ICU injury burden scores were strongly associated levels and scores at other time epochs.\u003c/p\u003e\n\u003cp\u003eNfL means at all time points were associated, in univariate regression, with 6-months GOSE, with higher NfL levels being associated with unfavorable outcome (\u003cstrong\u003e\u003cem\u003eTable 2A\u003c/em\u003e\u003c/strong\u003e). When adjusting for the ICU injury burden at the corresponding time point (\u003cstrong\u003e\u003cem\u003eTable 2B\u003c/em\u003e\u003c/strong\u003e), the NfL mean relationships to outcome were variably attenuated, while ICU injury burden scores were significantly related to outcome (p\u0026lt;0.05 all comparisons). \u0026nbsp;Here, Day 0 mean NfL was no longer significant in relationship to GOSE when Day 0 ICU injury burden was introduced (p=0.2497); however, Day 1-5 and Day 6-10 remained significant (p=0.0362 and p=0.0223, respectively). Together, the discriminative performance improved when ICU injury burden score was added to outcome models that included mean NfL levels. Each time epoch model showed a significant improvement in AUC (NfL only model vs. NfL + ICU injury burden model) when comparing ROC curves using Delong’s Test (p\u0026lt;0.05 all comparisons). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn preparation for multivariate model development, demographic (age, sex) and clinical injury factors (ER GCS, best in 24h GCS, CT burden) were investigated for NfL TRAJ and mean NfL measured over different time bins, with ICU injury burden and global outcome modeled as dependent variables (\u003cstrong\u003e\u003cem\u003eTable 3\u003c/em\u003e\u003c/strong\u003e). Older age was associated with higher Day 0 ICU injury burden and increased odds of unfavorable outcome. Injury factors (ER and best in 24h GCS, CT burden) were associated with both NfL TRAJ and unfavorable outcomes. Those with more severe injuries had an increased odds of having high NfL TRAJ profiles over the first 10 days post-injury and unfavorable 6-months outcome. CT burden was the factor most strongly associated with all ICU injury burden time epochs (p\u0026lt;0.0001 all comparisons).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNfL relationship to outcome in the context of ICU clinical course and demographic factors\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMultivariate logistic regression models were developed to examine relationships between the pattern of serum NfL levels over time and outcome, after adjusting for other injury severity variables known to be associated with outcome (age, sex, best in 24hr GCS and CT burden score). The baseline characteristic model is shown in \u003cstrong\u003e\u003cem\u003eTable 4A,\u003c/em\u003e\u0026nbsp;\u003c/strong\u003ewith NfL TRAJ and ICU injury burden added incrementally to demonstrate the relative influence of model factors and the change in outcome discrimination by AUC (from 0.787 to 0.803 and 0.819, respectively). Here, lower GCS (p=0.0104) and greater CT burden (p=0.004) were associated with increased odds of unfavorable GOSE (\u003cstrong\u003e\u003cem\u003eTable 4A\u003c/em\u003e\u003c/strong\u003e). When adding Day 0-10 NfL TRAJ to the model, these relationships were attenuated (GCS: p=0.0241, CT burden score: p=0.004) yet remained significant. There was a trend level association with high NfL TRAJ membership being associated with 2.57 increased odds of unfavorable GOSE versus the low TRAJ membership (\u003cstrong\u003e\u003cem\u003eTable 4B\u003c/em\u003e\u003c/strong\u003e). Lastly, when factoring in Day 0-10 ICU burden into the model (\u003cstrong\u003e\u003cem\u003eTable 4C\u003c/em\u003e\u003c/strong\u003e), the CT burden and NfL TRAJ relationships were fully attenuated (p\u0026gt;0.1), and the GCS association with outcome was attenuated to a lesser degree, remaining significantly associated with outcome (p=0.0106). For every unit increase in Day 0-10 ICU burden, there was 2.24 increased odds of unfavorable GOSE (p=0.0447). The AUC increased with the addition of NfL TRAJ and ICU injury burden; however, pairwise comparisons of ROC curves using DeLong’s test revealed no statistically significant differences in AUC between models (all p-values \u0026gt; 0.23), indicating similar discriminative performance.\u003c/p\u003e\n\u003cp\u003eThe dynamic role of ICU burden as a mediator of NfL associations over time with 6-months outcome was tested (\u003cstrong\u003e\u003cem\u003eFigure 3\u003c/em\u003e\u003c/strong\u003e) while adjusting for age and sex as covariates. The mediating effects of ICU burden on day 0 mean and day 1-5 mean NfL relationship to GOSE scores were tested subsequent ICU burden time epochs (\u003cstrong\u003e\u003cem\u003eFigure 3A\u003c/em\u003e\u003c/strong\u003e), and Day 6-10 ICU burden was assessed as a mediator for the day 1-5 NfL mean association with outcome (\u003cstrong\u003e\u003cem\u003eFigure 3B\u003c/em\u003e\u003c/strong\u003e). Day 1-5 ICU injury burden mediated 28% of the day 0 NfL mean association with unfavorable GOSE. Similarly, Day 6-10 ICU burden mediated 39.9% of the day 1-5 NfL mean association with unfavorable GOSE; however, the direct effect remained trend-level (p=0.0861). \u003cstrong\u003e\u003cem\u003eFigure 3C\u003c/em\u003e\u003c/strong\u003e demonstrates Day 0-10 ICU injury burden as a mediator of the NfL TRAJ relationship to GOSE. Here, the overall burden score mediated 28.9% of the NfL TRAJ association with GOSE resulting in a trend-level direct effect (p=0.0501). Together these models suggest that ICU injury burden is a partial mediator of the NfL biomarker associations with global outcome.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study highlights temporal and overall associations between NfL and ICU physiological parameters over the first 10 days after msTBI, and their collective impact on global outcome, while considering initial brain injury. The main strengths of the study include the serial biomarker measurements over the first 10 days post-injury along with the detailed clinical, ICU and radiological variables available for analyses.\u003c/p\u003e\u003cp\u003ePrevious studies have examined serial NfL measurements post-TBI, correlating increasing NfL serum or CSF levels over time with worse clinical outcome. Nimer et al\u003csup\u003e29\u003c/sup\u003e studied 182 patients with all TBI severities and collected 1 to 3 samples per patient over the course of 2 weeks. NfL levels showed an upward trend over time and correlated to outcome. Shahim et al\u003csup\u003e10\u003c/sup\u003e, evaluated 72 severe TBI patients where daily NfL measurements were obtained from day 0 to 12 post-injury. NfL levels were increased at admission, continued to rise daily, and also were associated with clinical outcome. In a longer time-course study we showed that serum NfL levels rise from the acute phase and peak at 30 days post-injury and both acute serum and CSF NfL levels discriminated 6- and 12-months outcome\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. While that study data included a portion of this cohort\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e, we similarly highlight that serum NfL levels keep rising in the acute phase post-TBI. This linear increase in NfL likely reflects ongoing axonal injury after the initial insult and the importance of serial measurements assessing longitudinal biomarker patterns rather than single time point measurement strategies.\u003c/p\u003e\u003cp\u003eThe TRAJ analysis approach\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e demonstrated temporal NfL dynamics by identifying two groups of individuals with unique serum NfL pattern over the first 10 days post-injury. A major advantage of TRAJ is that it is a clustering algorithm for assessing longitudinal patterns rather than single-point, averaged, or peak biomarker level for diagnostic or prognostic models. Its capacity for patient clustering for potential TBI patient subtype identification has been demonstrated for a variety of CNS derived\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e,\u003cspan additionalcitationids=\"CR31 CR32\" citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e and systemic biomarkers\u003csup\u003e\u003cspan additionalcitationids=\"CR35 CR36 CR37\" citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e. Validation studies are required to address the minimum time needed for patient assigned to a TRAJ group in real time based on their initial NfL levels; however, this methodology, and the dynamic relationships between NFL and ICU burden, shows potential utility for early clinical and prognostic management.\u003c/p\u003e\u003cp\u003eNfL levels demonstrated a linear increase over the first 10 days post-injury, contrary to GFAP, which presented with an early peak followed by variable decay patterns over the acute period\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. Rising NfL elevations over time likely reflect higher degree of evolving secondary axonal injury, as NfL means are closely associated with ICU injury burden over the full 10-day time course post-injury. The regression-based cross lag analysis showed mean NfL and ICU injury burden scores were significantly related to each other \u003cem\u003ewithin\u003c/em\u003e time epoch from day 0 (Day 0 NfL to Day 0 ICU injury burden) and \u003cem\u003eacross\u003c/em\u003e subsequent time epochs (day 0 to Day 1\u0026ndash;5, and Day 1\u0026ndash;5 to Day 6\u0026ndash;10). In contrast, Day 0 GFAP wasn\u0026rsquo;t associated with Day 0 ICU injury burden when using a similar analysis construct, highlighting the strong correlation of NfL with initial injury severity.\u003c/p\u003e\u003cp\u003eNfL levels were strongly correlated with GCS and CT burden. NfL means also positively correlated with both initial GCS, and best in 24h GCS, across all time epochs, with CT burden scores primarily associated with day 0 and day 1\u0026ndash;5 NfL levels. Similarly, higher NfL TRAJ groups were significantly associated with worse GCS and CT burden. This contrasts with the GFAP analysis\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e, where the CT characteristics but not the initial GCS and baseline injury severity factors were associated to the GFAP TRAJ group. Notably, other studies have shown GFAP levels within the first 24 hours correlated with admission GCS\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e. Both GFAP and NfL rise with aging in the general population\u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e,\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e. We found no age differences between \u003cem\u003elow\u003c/em\u003e and \u003cem\u003ehigh\u003c/em\u003e TRAJ groups. This work contrasts with our GFAP findings, where individuals in the high TRAJ GFAP group were significantly older\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eTo further evaluate the NfL association with ICU factors and outcome, age and sex were introduced as covariates in mediation analysis. GCS and CT burden scores were excluded due to associations between NfL TRAJ, ICU injury burden and GOSE, allowing NfL to be treated as the biological readout of initial injury severity and allowing it to be effectively modeled as a temporally dynamic variable in pathway analysis without the absence of confounding covariates with shared variance. Mediation analyses suggested that the ICU injury burden is a partial mediator, indicating that NfL biomarker associations with functional outcome occur via secondary CNS physiological injury response in response to neurocritical illness.\u003c/p\u003e\u003cp\u003eTaken together, these data suggest NfL is a robust blood-based biomarker representation of initial neurological injury severity, and interestingly, tracks closely with ICU factor time course. Independent of these correlative factors, higher NfL TRAJ and mean NfL levels measurement over the 10-day time epochs were each significantly associated with worse outcome. When introducing ICU injury burden into the modeling framework, these relationships were attenuated, suggesting ICU burden is in the causal pathway explaining a significant amount of the relationship between acute NfL levels and global outcome. These findings align with prior studies that have confirmed elevated NfL after a TBI of any severity is associated with degree of axonal injury\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e,\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e, and thus worse outcome at 6 months; however, the fact that acute ICU appears to have a causal effect on outcome suggests that there may be modifiable targets within ICU care best practice pathways where serial NfL monitoring may be a useful adjunct to clinical management and assessing clinical treatment effectiveness.\u003c/p\u003e\u003cp\u003eTo the best of our knowledge this is the first time NfL has been linked with acute ICU physiological parameters and acute treatment. Patients in the \u003cem\u003ehigh\u003c/em\u003e TRAJ group had more frequent refractory intracranial hypertension and cerebral hypoperfusion requiring advanced and invasive treatments. These findings build on previous work linking NfL to axonal injury and outcome post-TBI by providing an additional lens through ICU-related physiology to present opportunities for acute management and treatment.\u003c/p\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003eLimitations\u003c/h2\u003e\u003cp\u003eOur study has some limitations. Our sample size is relatively small, representing a single Level 1 trauma center\u0026rsquo;s experience. The clinical information available was exported from EMR and collected prospectively as part of a previous treatment trial. We did not assess the impact of each ICP treatment component on NfL levels, as in clinical practice multiple treatment measures are done simultaneously. Future larger, multicenter trials are necessary to validate the NfL role in acute ICU setting. Samples were only collected daily up to day 10, with no chronic samples available. However, for the purposes of this study, having serial samples in the first 10 days of ICU stay was sufficient to show the importance of longitudinal biomarker patterns in their relationship with ICU metrics over time. This work presents novel approaches to handling CT injury profiles and longitudinal ICU factors in time-specific load score derivations. However, the granular time course data and the presence of multicollinearity among NfL, clinical factors, and outcome limit the effectiveness of traditional regression approaches and may necessitate more sophisticated analytical techniques (e.g. machine learning) in larger cohorts to establish NFL as a biomarker for real-time clinical decision making. Future research with more frequent serial measurements over a longer time course, with multiple biomarker inclusion and advanced analysis planning, would further refine the role of NfL in individualized ICU care and prognostication, and provide treatment trial readouts for monitoring effectiveness.\u003c/p\u003e\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eSerum NfL levels correlate with worse clinical outcomes after msTBI and are associated with ICU clinical course. The linear increase of NfL levels and strong correlation with ICU burden and 6-months outcome provide information about the association with secondary brain injury following msTBI. Future research using a high-resolution sampling approach in assessing extended longitudinal patterns, as well as multi-marker panels for treatment triaging and clinical outcome is necessary in preparation for large scale multisite trials focused on assessing complex biological patterns and relationships to CNS physiology co-occurring critical illness associated with msTBI.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors contributions\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eConceptualization: \u0026nbsp;ST, CR, LM, KKW, AKW\u003c/p\u003e\n\u003cp\u003eData collection: CR, GC, KKW\u003c/p\u003e\n\u003cp\u003eData visualization, analysis, and interpretation: ST, LM, CR, DB, KKW, AKW\u003c/p\u003e\n\u003cp\u003eManuscript writing: ST, LM, CR, AKW\u003c/p\u003e\n\u003cp\u003eManuscript review and critical revision: CR, DB, JW, DL, FK, GC, RR, KKW, AKW\u003c/p\u003e\n\u003cp\u003eFunding: CR, JW, RR, KKW AKW\u003c/p\u003e\n\u003cp\u003eAll coauthors have seen and agree with the contents of the manuscript; the manuscript complies with all instructions to authors; requirements for authorship have been met; each author believes that the manuscript represents honest work; the STROBE checklist was used. This work has not been published elsewhere and is not under consideration by another journal and is not redundant with previous work. We used archived blood samples that were originally collected as part of our clinical trial of erythropoietin (clinicaltrials.gov NCT00313716). Patients were offered the option to have research blood samples banked for future studies analyses after clinical trial analysis were completion. Baylor College of Medicine Institutional Review Board approved waived consent for the use of these archived, de-identified blood samples (protocol H-44131).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e This project was funded by Department of Defense grant W81XWH-19-2-0012 (samples analysis, data analysis) and Department of Defense grant Gap-Based Milieu Biomarkers for Traumatic Brain Injury (GAMBIT- TBI) W81XWH2211089 (data analysis, manuscript preparation). The clinical trial of erythropoietin that originally collected the samples used in this study was pre-registered at clinicaltrials.gov (NCT00313716). Further support received from VA research grant IK2RX002490 and the Brain Rehabilitation Research Center, Malcom Randall VAMCGainesville FL.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclosures\u003c/strong\u003e: KKW is a share-holder of Gryphon Bio, Inc. The other authors have no relevant disclosures related to this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eTaylor CA, Bell JM, Breiding MJ, Xu L. Traumatic Brain Injury-Related Emergency Department Visits, Hospitalizations, and Deaths - United States, 2007 and 2013. MMWR Surveill Summ 2017;66(9):1-16. DOI: 10.15585/mmwr.ss6609a1.\u003c/li\u003e\n\u003cli\u003eHyder AA, Wunderlich CA, Puvanachandra P, Gururaj G, Kobusingye OC. The impact of traumatic brain injuries: a global perspective. NeuroRehabilitation 2007;22(5):341-53. (https://www.ncbi.nlm.nih.gov/pubmed/18162698).\u003c/li\u003e\n\u003cli\u003eMaas AIR, Menon DK, Manley GT, et al. Traumatic brain injury: progress and challenges in prevention, clinical care, and research. Lancet Neurol 2022;21(11):1004-1060. DOI: 10.1016/S1474-4422(22)00309-X.\u003c/li\u003e\n\u003cli\u003eShahim P, Politis A, van der Merwe A, et al. Time course and diagnostic utility of NfL, tau, GFAP, and UCH-L1 in subacute and chronic TBI. Neurology 2020;95(6):e623-e636. DOI: 10.1212/WNL.0000000000009985.\u003c/li\u003e\n\u003cli\u003eMouhieddine TH, El Houjeiri L, Sabra M, Hayes RL, Mondello S. CNS Trauma Biomarkers and Surrogate Endpoints Pipeline from Bench to Bedside: A Translational Perspective. In: Kobeissy FH, ed. Brain Neurotrauma: Molecular, Neuropsychological, and Rehabilitation Aspects. Boca Raton (FL)2015.\u003c/li\u003e\n\u003cli\u003eMondello S, Hayes RL. Biomarkers. Handb Clin Neurol 2015;127:245-65. DOI: 10.1016/B978-0-444-52892-6.00016-7.\u003c/li\u003e\n\u003cli\u003eNielson JL, Cooper SR, Yue JK, et al. Uncovering precision phenotype-biomarker associations in traumatic brain injury using topological data analysis. PLoS One 2017;12(3):e0169490. DOI: 10.1371/journal.pone.0169490.\u003c/li\u003e\n\u003cli\u003eKorley FK, Jain S, Sun X, et al. Prognostic value of day-of-injury plasma GFAP and UCH-L1 concentrations for predicting functional recovery after traumatic brain injury in patients from the US TRACK-TBI cohort: an observational cohort study. Lancet Neurol 2022;21(9):803-813. DOI: 10.1016/S1474-4422(22)00256-3.\u003c/li\u003e\n\u003cli\u003eBazarian JJ, Biberthaler P, Welch RD, et al. Serum GFAP and UCH-L1 for prediction of absence of intracranial injuries on head CT (ALERT-TBI): a multicentre observational study. Lancet Neurol 2018;17(9):782-789. DOI: 10.1016/S1474-4422(18)30231-X.\u003c/li\u003e\n\u003cli\u003eShahim P, Politis A, van der Merwe A, et al. Neurofilament light as a biomarker in traumatic brain injury. Neurology 2020;95(6):e610-e622. DOI: 10.1212/WNL.0000000000009983.\u003c/li\u003e\n\u003cli\u003eGao W, Zhang Z, Lv X, et al. Neurofilament light chain level in traumatic brain injury: A system review and meta-analysis. Medicine (Baltimore) 2020;99(38):e22363. DOI: 10.1097/MD.0000000000022363.\u003c/li\u003e\n\u003cli\u003eYuan A, Rao MV, Veeranna, Nixon RA. Neurofilaments and Neurofilament Proteins in Health and Disease. Cold Spring Harb Perspect Biol 2017;9(4). DOI: 10.1101/cshperspect.a018309.\u003c/li\u003e\n\u003cli\u003eGaetani L, Blennow K, Calabresi P, Di Filippo M, Parnetti L, Zetterberg H. Neurofilament light chain as a biomarker in neurological disorders. J Neurol Neurosurg Psychiatry 2019;90(8):870-881. DOI: 10.1136/jnnp-2018-320106.\u003c/li\u003e\n\u003cli\u003eMattsson N, Cullen NC, Andreasson U, Zetterberg H, Blennow K. Association Between Longitudinal Plasma Neurofilament Light and Neurodegeneration in Patients With Alzheimer Disease. JAMA Neurol 2019;76(7):791-799. DOI: 10.1001/jamaneurol.2019.0765.\u003c/li\u003e\n\u003cli\u003eBenedet AL, Ashton NJ, Pascoal TA, et al. Plasma neurofilament light associates with Alzheimer\u0026apos;s disease metabolic decline in amyloid-positive individuals. Alzheimers Dement (Amst) 2019;11:679-689. DOI: 10.1016/j.dadm.2019.08.002.\u003c/li\u003e\n\u003cli\u003eRohrer JD, Woollacott IO, Dick KM, et al. Serum neurofilament light chain protein is a measure of disease intensity in frontotemporal dementia. Neurology 2016;87(13):1329-36. DOI: 10.1212/WNL.0000000000003154.\u003c/li\u003e\n\u003cli\u003eOlsson B, Lautner R, Andreasson U, et al. CSF and blood biomarkers for the diagnosis of Alzheimer\u0026apos;s disease: a systematic review and meta-analysis. Lancet Neurol 2016;15(7):673-684. DOI: 10.1016/S1474-4422(16)00070-3.\u003c/li\u003e\n\u003cli\u003eLjungqvist J, Zetterberg H, Mitsis M, Blennow K, Skoglund T. Serum Neurofilament Light Protein as a Marker for Diffuse Axonal Injury: Results from a Case Series Study. J Neurotrauma 2017;34(5):1124-1127. DOI: 10.1089/neu.2016.4496.\u003c/li\u003e\n\u003cli\u003eShahim P, Pham DL, van der Merwe AJ, et al. Serum NfL and GFAP as biomarkers of progressive neurodegeneration in TBI. Alzheimers Dement 2024;20(7):4663-4676. DOI: 10.1002/alz.13898.\u003c/li\u003e\n\u003cli\u003eTrifilio E, Bottari S, McQuillan LE, et al. Temporal Profile of Serum Neurofilament Light (NF-L) and Heavy (pNF-H) Level Associations With 6-Month Cognitive Performance in Patients With Moderate-Severe Traumatic Brain Injury. J Head Trauma Rehabil 2024. DOI: 10.1097/HTR.0000000000000932.\u003c/li\u003e\n\u003cli\u003eShahim P, Gren M, Liman V, et al. Serum neurofilament light protein predicts clinical outcome in traumatic brain injury. Sci Rep 2016;6:36791. DOI: 10.1038/srep36791.\u003c/li\u003e\n\u003cli\u003eGraham NSN, Zimmerman KA, Moro F, et al. Axonal marker neurofilament light predicts long-term outcomes and progressive neurodegeneration after traumatic brain injury. Sci Transl Med 2021;13(613):eabg9922. DOI: 10.1126/scitranslmed.abg9922.\u003c/li\u003e\n\u003cli\u003eWang KKW, Barton DJ, McQuillan LE, et al. Parallel Cerebrospinal Fluid and Serum Temporal Profile Assessment of Axonal Injury Biomarkers Neurofilament-Light Chain and Phosphorylated Neurofilament-Heavy Chain: Associations With Patient Outcome in Moderate-Severe Traumatic Brain Injury. J Neurotrauma 2024;41(13-14):1609-1627. DOI: 10.1089/neu.2023.0449.\u003c/li\u003e\n\u003cli\u003eRobertson CS, Martinez FS, McQuillan LE, et al. Serial Measurements of Serum Glial Fibrillary Acidic Protein in Moderate-Severe Traumatic Brain Injury: Potential Utility in Providing Insights into Secondary Insults and Long-Term Outcome. J Neurotrauma 2024;41(1-2):73-90. DOI: 10.1089/neu.2023.0111.\u003c/li\u003e\n\u003cli\u003eSteyerberg EW, Mushkudiani N, Perel P, et al. Predicting outcome after traumatic brain injury: development and international validation of prognostic scores based on admission characteristics. PLoS Med 2008;5(8):e165; discussion e165. DOI: 10.1371/journal.pmed.0050165.\u003c/li\u003e\n\u003cli\u003eFriedman J, Hastie T, Tibshirani R. Regularization Paths for Generalized Linear Models via Coordinate Descent. J Stat Softw 2010;33(1):1-22. (https://www.ncbi.nlm.nih.gov/pubmed/20808728).\u003c/li\u003e\n\u003cli\u003eNiyonkuru C, Wagner AK, Ozawa H, Amin K, Goyal A, Fabio A. Group-based trajectory analysis applications for prognostic biomarker model development in severe TBI: a practical example. J Neurotrauma 2013;30(11):938-45. DOI: 10.1089/neu.2012.2578.\u003c/li\u003e\n\u003cli\u003eBaron RM, Kenny DA. The moderator-mediator variable distinction in social psychological research: conceptual, strategic, and statistical considerations. J Pers Soc Psychol 1986;51(6):1173-82. DOI: 10.1037//0022-3514.51.6.1173.\u003c/li\u003e\n\u003cli\u003eAl Nimer F, Thelin E, Nystrom H, et al. Comparative Assessment of the Prognostic Value of Biomarkers in Traumatic Brain Injury Reveals an Independent Role for Serum Levels of Neurofilament Light. PLoS One 2015;10(7):e0132177. DOI: 10.1371/journal.pone.0132177.\u003c/li\u003e\n\u003cli\u003eRubenstein R, McQuillan L, Wang KKW, et al. Temporal Profiles of P-Tau, T-Tau, and P-Tau:Tau Ratios in Cerebrospinal Fluid and Blood from Moderate-Severe Traumatic Brain Injury Patients and Relationship to 6-12 Month Global Outcomes. J Neurotrauma 2024;41(3-4):369-392. DOI: 10.1089/neu.2022.0479.\u003c/li\u003e\n\u003cli\u003eKumar RG, Diamond ML, Boles JA, et al. Acute CSF interleukin-6 trajectories after TBI: associations with neuroinflammation, polytrauma, and outcome. Brain Behav Immun 2015;45:253-62. DOI: 10.1016/j.bbi.2014.12.021.\u003c/li\u003e\n\u003cli\u003eWagner AK, Amin KB, Niyonkuru C, et al. CSF Bcl-2 and cytochrome C temporal profiles in outcome prediction for adults with severe TBI. J Cereb Blood Flow Metab 2011;31(9):1886-96. DOI: 10.1038/jcbfm.2011.31.\u003c/li\u003e\n\u003cli\u003eGoyal A, Failla MD, Niyonkuru C, et al. S100b as a prognostic biomarker in outcome prediction for patients with severe traumatic brain injury. J Neurotrauma 2013;30(11):946-57. DOI: 10.1089/neu.2012.2579.\u003c/li\u003e\n\u003cli\u003eMunoz MJ, Kumar RG, Oh BM, et al. Cerebrospinal Fluid Cortisol Mediates Brain-Derived Neurotrophic Factor Relationships to Mortality after Severe TBI: A Prospective Cohort Study. Front Mol Neurosci 2017;10:44. DOI: 10.3389/fnmol.2017.00044.\u003c/li\u003e\n\u003cli\u003eSantarsieri M, Kumar RG, Kochanek PM, Berga S, Wagner AK. Variable neuroendocrine-immune dysfunction in individuals with unfavorable outcome after severe traumatic brain injury. Brain Behav Immun 2015;45:15-27. DOI: 10.1016/j.bbi.2014.09.003.\u003c/li\u003e\n\u003cli\u003eSantarsieri M, Niyonkuru C, McCullough EH, et al. Cerebrospinal fluid cortisol and progesterone profiles and outcomes prognostication after severe traumatic brain injury. J Neurotrauma 2014;31(8):699-712. DOI: 10.1089/neu.2013.3177.\u003c/li\u003e\n\u003cli\u003eBarton DJ, Kumar RG, Schuster AA, Juengst SB, Oh BM, Wagner AK. Acute Cortisol Profile Associations With Cognitive Impairment After Severe Traumatic Brain Injury. Neurorehabil Neural Repair 2021;35(12):1088-1099. DOI: 10.1177/15459683211048771.\u003c/li\u003e\n\u003cli\u003eWagner AK, McCullough EH, Niyonkuru C, et al. Acute serum hormone levels: characterization and prognosis after severe traumatic brain injury. J Neurotrauma 2011;28(6):871-88. DOI: 10.1089/neu.2010.1586.\u003c/li\u003e\n\u003cli\u003eOkonkwo DO, Puffer RC, Puccio AM, et al. Point-of-Care Platform Blood Biomarker Testing of Glial Fibrillary Acidic Protein versus S100 Calcium-Binding Protein B for Prediction of Traumatic Brain Injuries: A Transforming Research and Clinical Knowledge in Traumatic Brain Injury Study. J Neurotrauma 2020;37(23):2460-2467. DOI: 10.1089/neu.2020.7140.\u003c/li\u003e\n\u003cli\u003eKhalil M, Pirpamer L, Hofer E, et al. Serum neurofilament light levels in normal aging and their association with morphologic brain changes. Nat Commun 2020;11(1):812. DOI: 10.1038/s41467-020-14612-6.\u003c/li\u003e\n\u003cli\u003eTybirk L, Hviid CVB, Knudsen CS, Parkner T. Serum GFAP - reference interval and preanalytical properties in Danish adults. Clin Chem Lab Med 2022;60(11):1830-1838. DOI: 10.1515/cclm-2022-0646.\u003c/li\u003e\n\u003cli\u003eDisanto G, Barro C, Benkert P, et al. Serum Neurofilament light: A biomarker of neuronal damage in multiple sclerosis. Ann Neurol 2017;81(6):857-870. DOI: 10.1002/ana.24954.\u003c/li\u003e\n\u003cli\u003eShahim P, Zetterberg H, Tegner Y, Blennow K. Serum neurofilament light as a biomarker for mild traumatic brain injury in contact sports. Neurology 2017;88(19):1788-1794. DOI: 10.1212/WNL.0000000000003912.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1A: NfL trajectory group comparisons \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 290px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL TRAJ GROUP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLow NfL\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHigh NfL\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003ep-value\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber of patients:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSex\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eMale\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eFemale\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 51\u003c/p\u003e\n \u003cp\u003e- 12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 28\u003c/p\u003e\n \u003cp\u003e- 6\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.86\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge (mean)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e- 32.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;34.6\u003c/p\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e0.6884\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdmission Motor GCS\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e1-3\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e4-6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 17 (27%)\u003c/p\u003e\n \u003cp\u003e- 46 (73%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 22 (64%)\u003c/p\u003e\n \u003cp\u003e- 12 (36%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.0005\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdmission Sum GCS\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e3-5\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e6-8\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e\u0026gt;8\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 15 (24%)\u003c/p\u003e\n \u003cp\u003e- 30 (48%)\u003c/p\u003e\n \u003cp\u003e- 18 (28%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 21 (61%)\u003c/p\u003e\n \u003cp\u003e- 7 (21%)\u003c/p\u003e\n \u003cp\u003e- 6 (18%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.0009\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGCS (best in 24h), median [IQR]\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e- 6 [4-7]\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e- 4.5 [4-7]\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u003cem\u003e0.0646\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePupils:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eBoth reactive\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eOne reactive\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eNeither reactive\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 46 (73%)\u003c/p\u003e\n \u003cp\u003e- 6 (10%)\u003c/p\u003e\n \u003cp\u003e- 11 (17%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 17 (50%)\u003c/p\u003e\n \u003cp\u003e- 6 (18%)\u003c/p\u003e\n \u003cp\u003e- 11 (32%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre hospital hypotension\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eAbsent\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003ePresent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 59 (94%)\u003c/p\u003e\n \u003cp\u003e- 4 (6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 27 (80%)\u003c/p\u003e\n \u003cp\u003e- 7 (20%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.046\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre hospital Hypoxia\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eAbsent\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003ePresent\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 51 (81%)\u003c/p\u003e\n \u003cp\u003e- 12 (19%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 22 (65%)\u003c/p\u003e\n \u003cp\u003e- 12 (35%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e0.08\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdmission Marshall CT\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eDiffuse injury 2\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eDiffuse injury 3\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eEvacuated Mass Lesion\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eUnevacuated Mass Lesion\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 31\u003c/p\u003e\n \u003cp\u003e- 14\u003c/p\u003e\n \u003cp\u003e- 16\u003c/p\u003e\n \u003cp\u003e- 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 8\u003c/p\u003e\n \u003cp\u003e- 13\u003c/p\u003e\n \u003cp\u003e- 12\u003c/p\u003e\n \u003cp\u003e- 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e0.09\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSubarachnoid Hemorrhage\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003ePresent\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eAbsent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 41 (65%)\u003c/p\u003e\n \u003cp\u003e- 22 (35%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 27 (79%)\u003c/p\u003e\n \u003cp\u003e- 7 (21%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBasal Cisterns\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eNormal\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eCompressed/absent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 37 (59%)\u003c/p\u003e\n \u003cp\u003e- 26 (41%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 12 (35%)\u003c/p\u003e\n \u003cp\u003e- 22 (65%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.03\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMidline shift\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eNone\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e\u0026lt; 5 mm\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e5-15 mm\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003e\u0026gt;15 mm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 44 (70%)\u003c/p\u003e\n \u003cp\u003e- 8 (13%)\u003c/p\u003e\n \u003cp\u003e- 9 (14%)\u003c/p\u003e\n \u003cp\u003e- 2 (3%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 16 (47%)\u003c/p\u003e\n \u003cp\u003e- 6 (18%)\u003c/p\u003e\n \u003cp\u003e- 10 (29%)\u003c/p\u003e\n \u003cp\u003e- 2 (6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDiffuse Brain Injury (DBI)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003ePresent\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eAbsent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 45 (71.4%)\u003c/p\u003e\n \u003cp\u003e- 18 (28.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 22 (64.7%)\u003c/p\u003e\n \u003cp\u003e- 12 (35.3%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e0.4943\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCT Burden, median [IQR]\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e0.51 [0.5-1.62]\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e1.62 [0.87-1.97]\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0086\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVentricles\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eNormal\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eSmall\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eEnlarged\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 29 (46%)\u003c/p\u003e\n \u003cp\u003e- 33 (52%)\u003c/p\u003e\n \u003cp\u003e- 1 (2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 12 (35%)\u003c/p\u003e\n \u003cp\u003e- 22 (65%)\u003c/p\u003e\n \u003cp\u003e- 0 (0%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.53\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIMPACT risk of poor outcome (mean)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e0.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIMPACT risk of mortality (mean)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGOSE at 6 mo\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eGR/MD\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eSD/V/D\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eMissing/lost to f/u\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 30 (48%)\u003c/p\u003e\n \u003cp\u003e- 30 (48%)\u003c/p\u003e\n \u003cp\u003e- 3 (4%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 4 (12%)\u003c/p\u003e\n \u003cp\u003e- 26 (76%)\u003c/p\u003e\n \u003cp\u003e- 4 (12%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMortality at 6 mo\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eSurvived\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eDied\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e- \u003cstrong\u003eMissing/lost to f/u\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 57 (90%)\u003c/p\u003e\n \u003cp\u003e- 3 (4%)\u003c/p\u003e\n \u003cp\u003e- 3 (4%)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 21 (62%)\u003c/p\u003e\n \u003cp\u003e- 9 (26%)\u003c/p\u003e\n \u003cp\u003e- 4 (12%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eBold denotes p\u0026le;0.05; italic denotes p\u0026le;0.1. GCS: Glasgow Coma Scale. GOSE GR/MD/SD/V/D: Good Recovery/Moderate Disability/Severe Disability/Vegetative State/Dead. \u0026nbsp;IQR: interquartile range. IMPACT: International Mission for Prognosis and Analysis of Clinical Trials in TBI.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1B: Secondary injury measures for patients in the two NfL trajectory groups\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 292px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL TRAJ GROUP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003eLow NfL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003eHigh NfL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003ep-value\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eNumber of patients\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eAverage ICP (mean, mmHg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e14.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e18.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eICP\u003c/p\u003e\n \u003cp\u003e- Never \u0026gt;25\u003c/p\u003e\n \u003cp\u003e- \u0026gt; 25mmHg at least once\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 16 (25%)\u003c/p\u003e\n \u003cp\u003e- 47 (75%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 10 (29%)\u003c/p\u003e\n \u003cp\u003e- 24 (71%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.81\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eICP duration \u0026gt; 25 mmHg (h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e9.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e20.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eICP\u003c/p\u003e\n \u003cp\u003e- Never \u0026gt; 30\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- \u0026gt; 30 mmHg at least once\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 25 ( 40%)\u003c/p\u003e\n \u003cp\u003e- 38 (60%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 15 (44%)\u003c/p\u003e\n \u003cp\u003e- 19 (56%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eICP duration \u0026gt; 30 mmHg (h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e2.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e9.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0069\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eRequired mannitol:\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- No\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- Yes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 24 (38%)\u003c/p\u003e\n \u003cp\u003e- 39 (62%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 11 (33%)\u003c/p\u003e\n \u003cp\u003e- 23 (67%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.66\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eRequired barbiturate coma:\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- No\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- Yes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 57 (91%)\u003c/p\u003e\n \u003cp\u003e- 6 (9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 24 (71%)\u003c/p\u003e\n \u003cp\u003e- 10 (29%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eRequired decompressive craniectomy\u003c/p\u003e\n \u003cp\u003e- No\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- Yes\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 46 (73%)\u003c/p\u003e\n \u003cp\u003e- 17 (27%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 17 (50%)\u003c/p\u003e\n \u003cp\u003e- 17 (50%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.02\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eAverage MAP (mmHg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e90.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e91.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e0.60\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eMAP:\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- Never \u0026lt; 70 mmHg\u003c/p\u003e\n \u003cp\u003e- \u0026lt; 70 mmHg at least once\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 26 (41%)\u003c/p\u003e\n \u003cp\u003e- 37 (59%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 8 (24%)\u003c/p\u003e\n \u003cp\u003e- 26 (76%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eMAP duration \u0026lt; 70 mmHg (h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e3.4\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e4.7\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eAverage CPP (mmHg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e75.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e72.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cem\u003e0.06\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eCPP:\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- Never \u0026lt;50mmHg\u003c/p\u003e\n \u003cp\u003e- \u0026lt; 50mmHg at least once\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 23 (37%)\u003c/p\u003e\n \u003cp\u003e- 40 (63%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 9 (26%)\u003c/p\u003e\n \u003cp\u003e- 25 (74%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eCPP duration \u0026lt;50mmHg (h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e2.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e7.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eAverage PbtO\u003csub\u003e2\u003c/sub\u003e (mmHg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e33.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e26.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u003cem\u003e0.05\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003ePbtO\u003csub\u003e2\u003c/sub\u003e :\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- Never \u0026lt;10 mmHg\u003c/p\u003e\n \u003cp\u003e- \u0026lt;10mmHg at least once\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 38 (62%)\u003c/p\u003e\n \u003cp\u003e- 23 (38%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 18 (55%)\u003c/p\u003e\n \u003cp\u003e- 15 (45%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003ePbtO\u003csub\u003e2\u003c/sub\u003e duration \u0026lt;10mmHg (mean, hours)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e4.75\u003c/p\u003e\n \u003cp\u003e(missing 2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e14.6\u003c/p\u003e\n \u003cp\u003e(missing 1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eAverage Sjv02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e76.93\u003c/p\u003e\n \u003cp\u003eMissing 14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e74.83\u003c/p\u003e\n \u003cp\u003eMissing 6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eSjvO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003cp\u003e- Never \u0026lt;50%\u003c/p\u003e\n \u003cp\u003e- \u0026lt;50% at least once\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 38 (78%)\u003c/p\u003e\n \u003cp\u003e- 11 (22%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 17 (61%)\u003c/p\u003e\n \u003cp\u003e- 11 (39%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eSjvO\u003csub\u003e2\u003c/sub\u003e duration \u0026lt;50% (h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e1.17\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eDelayed hematoma\u003c/p\u003e\n \u003cp\u003e- Absent\u003c/p\u003e\n \u003cp\u003e- Present\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 44 (70%)\u003c/p\u003e\n \u003cp\u003e- 19 (30%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 22 (65%)\u003c/p\u003e\n \u003cp\u003e- 12 (35%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e0.65\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 182px;\"\u003e\n \u003cp\u003eWorst Marshall CT\u003c/p\u003e\n \u003cp\u003e- Diffuse injury 2\u003c/p\u003e\n \u003cp\u003e- Diffuse injury 3\u003c/p\u003e\n \u003cp\u003e- Diffuse injury 4\u003c/p\u003e\n \u003cp\u003e- Evacuated Mass Lesion\u003c/p\u003e\n \u003cp\u003e- Unevacuated Mass Lesion\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 154px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 28\u003c/p\u003e\n \u003cp\u003e- 14\u003c/p\u003e\n \u003cp\u003e- 1\u003c/p\u003e\n \u003cp\u003e- 17\u003c/p\u003e\n \u003cp\u003e- 3\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e- 7\u003c/p\u003e\n \u003cp\u003e- 13\u003c/p\u003e\n \u003cp\u003e- 1\u003c/p\u003e\n \u003cp\u003e- 11\u003c/p\u003e\n \u003cp\u003e- 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 103px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e0.0014\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eBold denotes p\u0026le;0.05; italic denotes p\u0026le;0.1.ICP: intracranial pressure. MAP: Mean arterial pressure. CPP: Cerebral perfusion pressure. PbtO2: Brain tissue oxygen partial pressure. SjvO2:\u0026nbsp;Jugular venous oxygen saturation. CT: Computed Tomography\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. GOSE outcome models of (A) serum NfL means by time epoch, and (B) both serum NfL mean and GOSE specific ICU injury burden score at all time epochs\u003c/strong\u003e. \u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"635\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 635px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e(A) NfL Mean \u0026rarr; 6-month GOSE outcome (individual regressions)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAUC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL Mean (day 0)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.007\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.001-1.014\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0326\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0.646\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL Mean (days 1-5)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.005\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.001-1.008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0108\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0.660\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL Mean (days 6-10)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.00-1.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0111\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0.664\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 635px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e (B) NfL Mean (Adjusted for ICU injury burden score) \u0026rarr; 6-month GOSE outcome \u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAUC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL (day 0)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e0.99-1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e0.2497\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0.779\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eICU injury burden (day 0)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e3.418\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.72-6.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAUC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL (days 1-5)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.0-1.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0362\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0.733\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eICU injury burden (days 1-5)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e3.814\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.35-10.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0013\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAUC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSerum NfL (days 6-10)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.0-1.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0223\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0.770\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 215px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eICU injury burden (days 6-10)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e2.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e1.25-7.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 137px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0141\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eEach row represents and individual univariate binary logistic regression in A) with the corresponding odds ratio (OR), 95% confidence interval (CI), p-value, and area under the curve (AUC). Each pair of NfL and ICU injury burden by time epoch represent an individual multivariate binary logistic regression in B) with the corresponding OR, CI, p-value, and AUCs. Model results odds of an unfavorable GOSE=1-4. \u0026nbsp;Bold denotes p\u0026le;0.05. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Individual regression models to A) 6-month GOSE outcome, B) Day 0-10 NfL TRAJ group, and C-F) ICU injury burden at all time epochs\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"750\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA) 6-month GOSE\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(dichotomized)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB) Day 0-10 NfL TRAJ\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC) Day 0 ICU injury burden\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD) Day 1-5 ICU injury burden\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eE) Day 6-10 ICU injury burden\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF) Day 0-10 ICU injury burden\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR [CI]\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR [CI]\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB (SE)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB (SE)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB (SE)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB (SE)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eAge\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e1.034 [1.0-1.07]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0382\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e1.012 [0.98-1.04]\u003c/p\u003e\n \u003cp\u003ep=0.4316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.01 (0.009)\u003c/p\u003e\n \u003cp\u003e\u003cem\u003ep=0.0889\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e0.0038 (0.004)\u003c/p\u003e\n \u003cp\u003ep=0.2896\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e0.0001 (0.004)\u003c/p\u003e\n \u003cp\u003ep=0.9860\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e0.001 (0.005)\u003c/p\u003e\n \u003cp\u003ep=0.8994\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e0.826 [0.29-2.34]\u003c/p\u003e\n \u003cp\u003ep=0.7179\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e1.098 [0.37-3.24]\u003c/p\u003e\n \u003cp\u003ep=0.8656\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e-0.54 (0.35)\u003c/p\u003e\n \u003cp\u003ep=0.1259\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e-0.088 (0.13)\u003c/p\u003e\n \u003cp\u003ep=0.4979\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e-0.15 (0.05)\u003c/p\u003e\n \u003cp\u003ep=0.3212\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e-0.19 (0.19)\u003c/p\u003e\n \u003cp\u003ep=0.3334\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eER GCS Sum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e0.74 [0.63-0.87]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0003\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e0.785 [0.663-0.93]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0049\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e-0.09 (0.05)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0454\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e-0.008 (0.02)\u003c/p\u003e\n \u003cp\u003ep=0.6365\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e-0.03 (0.02)\u003c/p\u003e\n \u003cp\u003ep=0.1274\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e-0.04 (0.03)\u003c/p\u003e\n \u003cp\u003ep=0.1681\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eGCS\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e(best in 24h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e0.725 [0.58-0.9]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0037\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e0.779 [0.62-0.98]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0357\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e-0.05 (0.06)\u003c/p\u003e\n \u003cp\u003ep=0.4336\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e-0.01 (0.02)\u003c/p\u003e\n \u003cp\u003ep=0.6485\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e-0.01 (0.03)\u003c/p\u003e\n \u003cp\u003ep=0.7074\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e-0.03 (0.04)\u003c/p\u003e\n \u003cp\u003ep=0.4540\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eCT burden\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 126px;\"\u003e\n \u003cp\u003e3.73 [1.99-6.98]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e2.24 [1.19-4.24]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep=0.0130\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 90px;\"\u003e\n \u003cp\u003e0.72 (0.16)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e0.34 (0.06)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e0.42 (0.06)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003e0.56 (0.08)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003ep\u0026lt;0.0001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eDay 0 ICU injury burden factors:\u0026nbsp;Duration (h) CPP \u0026lt;50, Duration (h) PbtO2 \u0026lt;10, Duration (h) ICP \u0026gt;25, mannitol dose required, and decompressive craniectomy and/or barbiturate coma dose required\u003c/p\u003e\n\u003cp\u003eDay 1-5 ICU injury burden factor: duration (h) PbtO2 \u0026lt;10, decompressive craniectomy and/or barbiturate coma dose required.\u003c/p\u003e\n\u003cp\u003eDay 6-10 ICU injury burden factors: mannitol dose required, decompressive craniectomy and/or barbiturate coma dose required\u003c/p\u003e\n\u003cp\u003eDay 0-10 ICU injury burden factors: duration (h) CPP\u0026lt;50, Duration (h) PbtO2\u0026lt;10, and decompressive craniectomy and/or barbiturate coma dose required.\u003c/p\u003e\n\u003cp\u003eBold denotes p\u0026le;0.05; italic denotes p\u0026le;0.1. GCS: Glasgow Coma Scale. CT: Computed Tomography. ICU: Intensive Care Unit. OR: Odds Ratio. \u0026nbsp;CI: Confidence interval. SE: Standard Error\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4. Covariate adjusted logistic regression models of dichotomized 6-month GOSE outcome.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"658\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e6-month GOSE\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(dichotomized)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 22.2823%;\"\u003e\n \u003col\u003e\n \u003cli\u003e\u003cstrong\u003eBaseline Characteristic Model (AUC=0.787)\u003c/strong\u003e\u003c/li\u003e\n \u003c/ol\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 22.0277%;\"\u003e\n \u003col start=\"2\"\u003e\n \u003cli\u003e\u003cstrong\u003eNfL Risk Model\u003c/strong\u003e\u003c/li\u003e\n \u003c/ol\u003e\n \u003cp\u003e\u003cstrong\u003e(AUC=0.803)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 18.6561%;\"\u003e\n \u003col start=\"100\"\u003e\n \u003cli\u003e\u003cstrong\u003eNfL Risk + ICU injury burden\u003c/strong\u003e\u003c/li\u003e\n \u003c/ol\u003e\n \u003cp\u003e\u003cstrong\u003eModel (AUC=0.819)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIndependent variables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003eAge\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e0.97-1.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e0.5852\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e1.008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e0.97-1.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e0.6828\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e1.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e0.97-1.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e0.4267\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003eGender (Male vs. Female)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e1.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e0.39-4.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e0.6217\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e1.303\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e0.36-4.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e0.6853\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e1.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e0.28-4.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e0.8921\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003eGCS, best in 24h\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e0.58-0.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0104\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e0.57-0.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0241\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e0.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e0.58-0.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0106\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003eCT burden score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e3.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e1.76-7.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e3.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e1.42-6.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0040\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e2.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e0.87-4.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.1007\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003eNfL risk TRAJ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e2.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e0.89-7.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.0823\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e1.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e0.65-5.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e0.2361\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 15.7886%;\"\u003e\n \u003cp\u003eOverall Day 0-10 ICU injury burden score (GOSE specific)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.2577%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.385%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.6397%;\"\u003e\n \u003cp\u003e2.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.767%;\"\u003e\n \u003cp\u003e1.02-4.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7.5123%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0447\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eBold denotes p\u0026le;0.05. Italic denotes p\u0026le;0.1. GCS: Glasgow Coma Scale. CT: Computed Tomography. ICU: Intensive Care Unit. OR: Odds Ratio. \u0026nbsp;CI: Confidence interval.\u0026nbsp;\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"neurocritical-care","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"neca","sideBox":"Learn more about [Neurocritical Care](http://link.springer.com/journal/12028)","snPcode":"12028","submissionUrl":"https://www.editorialmanager.com/neca/default2.aspx","title":"Neurocritical Care","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"traumatic brain injury, critical illness, intracranial pressure, cerebral perfusion pressure, brain tissue oxygenation, neurofilament light","lastPublishedDoi":"10.21203/rs.3.rs-7909088/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7909088/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eRecent studies demonstrate increased Neurofilament Light (NfL) levels across all types of Traumatic Brain Injury (TBI). We aimed to evaluate the relationship between acute temporal NfL with ICU physiology and outcome.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eStored serum from 97 patients with severe TBI were analyzed. Samples werecollected every 6h for the first 24h and then daily through day 10. Temporal NfL patterns were examined using group-based trajectory (TRAJ) modeling, and means were calculated over day 0, 1-5, 6-10 and 0-10. ICU injury burden scores were generated using ridge regression anchored to Glasgow Outcome Scale-Extended (GOSE) over the same time-bins. We used both univariable and multivariable regression to assess NfL TRAJ and means with respect to ICU injury burden, demographic data, clinical factors, and outcome. NfL TRAJ and day 0-10 ICU injury burden were added to a baseline model to evaluate outcome discrimination. Mediation analysis was applied to assess the mediation effect of ICU injury burden between NfL and GOSE.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003ePatients demonstrated increasing levels over the monitoring period. Sixty-three patients had low NfL temporal profile (low-TRAJ), and 34 patients had higher profile (high-TRAJ). Individuals in the high-TRAJ group had lower GCS (p=0.0009), more frequently required barbiturate coma (p=0.01) or decompressive surgery (p=0.02) and had worse 6-months outcome (p=0.001). NfL means and ICU injury burden both negatively correlated with GCS. Multivariate logistic regression showed a 2.24X increased odds of unfavorable GOSE per unit increase in ICU injury burden (p=0.0447). When modeled with ICU injury burden, the NfL high-TRAJ group relationship to outcome was attenuated, with ICU burden being a significant mediator of the NfL relationship to GOSE.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eOur results suggest that ICU injury burden contributes to serum NfL associations with GOSE. Temporal NfL profiles provide novel information about intracranial pathophysiology as a causal mediator of axonal injury burden following severe TBI.\u003c/p\u003e","manuscriptTitle":"Neurofilament Light Protein: a marker for injury severity, clinical course and outcome following moderate-severe Traumatic Brain Injury","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-13 07:51:28","doi":"10.21203/rs.3.rs-7909088/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-10-30T21:10:02+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-30T19:54:39+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Neurocritical Care","date":"2025-10-29T14:26:43+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-27T21:43:05+00:00","index":"","fulltext":""},{"type":"submitted","content":"Neurocritical Care","date":"2025-10-24T21:33:58+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"neurocritical-care","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"neca","sideBox":"Learn more about [Neurocritical Care](http://link.springer.com/journal/12028)","snPcode":"12028","submissionUrl":"https://www.editorialmanager.com/neca/default2.aspx","title":"Neurocritical Care","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"833640d5-6891-4f79-8335-3186b1f246f1","owner":[],"postedDate":"November 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-08T12:55:30+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-13 07:51:28","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7909088","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7909088","identity":"rs-7909088","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}