High-quality and robust protein quantification in large clinical/pharmaceutical cohorts with IonStar proteomics investigation.

Robust, reliable quantification of large sample cohorts is often essential for meaningful clinical or pharmaceutical proteomics investigations, but it is technically challenging. When analyzing very large numbers of samples, isotope labeling approaches may suffer from substantial batch effects, and even with label-free methods, it becomes evident that low-abundance proteins are not reliably measured owing to unsufficient reproducibility for quantification. The MS1-based quantitative proteomics pipeline IonStar was designed to address these challenges. IonStar is a label-free approach that takes advantage of the high sensitivity/selectivity attainable by ultrahigh-resolution (UHR)-MS1 acquisition (e.g., 120-240k full width at half maximum at m/z = 200) which is now widely available on ultrahigh-field Orbitrap instruments. By selectively and accurately procuring quantitative features of peptides within precisely defined, very narrow m/z windows corresponding to the UHR-MS1 resolution, the method minimizes co-eluted interferences and substantially enhances signal-to-noise ratio of low-abundance species by decreasing noise level. This feature results in high sensitivity, selectivity, accuracy and precision for quantification of low-abundance proteins, as well as fewer missing data and fewer false positives. This protocol also emphasizes the importance of well-controlled, robust experimental procedures to achieve high-quality quantification across a large cohort. It includes a surfactant cocktail-aided sample preparation procedure that achieves high/reproducible protein/peptide recoveries among many samples, and a trapping nano-liquid chromatography-mass spectrometry strategy for sensitive and reproducible acquisition of UHR-MS1 peptide signal robustly across a large cohort. Data processing and quality evaluation are illustrated using an example dataset ( http://proteomecentral.proteomexchange.org ), and example results from pharmaceutical project and one clinical project (patients with acute respiratory distress syndrome) are shown. The complete IonStar pipeline takes ~1-2 weeks for a sample cohort containing ~50-100 samples.

Pharmacological Characterization of a Novel Neuroactive Steroid Prodrug, NTS-104, and Its Neuroprotective Activity in Experimental Stroke Models.

BACKGROUND: Ischemic stroke affects about 700 000 patients per year in the United States, and to date, there are no effective pharmacological agents that promote recovery. Here, we studied the pharmacokinetics, pharmacodynamics, and efficacy of NTS-105, a novel neuroactive steroid, and NTS-104, a prodrug of NTS-105, in 2 models of ischemic stroke. METHODS: The pharmacodynamics and pharmacokinetics of NTS-104/105 were investigated in naive and stroke rats, and models of embolic and transient middle cerebral artery occlusion were used to investigate the dose-related effects of NTS-104. All rats were randomly assigned into the experimental groups, and all outcome measurements were performed blindly. RESULTS: Blood plasma and brain pharmacokinetic analysis revealed that NTS-104 rapidly converted to NTS-105, which reached peak concentration at ≈1 hour after dosing and distributed similarly to normal and ischemic brains. NTS-104 administration 4 hours after embolic middle cerebral artery occlusion led to a dose-dependent improvement of neurological outcomes and a dose-dependent reduction of infarct volumes relative to vehicle-treated animals. A single dose level study confirmed that NTS-104 administered 4 hours after transient middle cerebral artery occlusion was also neuroprotective. Quantitative ELISA revealed that NTS-104 treatment resulted in time- and dose-dependent changes in AKT activation and cytokine levels within the ischemic brain, which included reductions of IL-6, VEGF, ICAM-1, IL-1ß, MCP-1, RAGE, and GM-CSF. Time- and dose-dependent reductions in IL-6 and GM-CSF were also observed in the plasma along with an elevation of galectin-1. CONCLUSIONS: NTS-104 is a novel prodrug that converts to a novel neuroactive steroid, NTS-105, which improves functional outcomes in experimental ischemic stroke models.

Characteristics of ocular trauma in the United States / Características de trauma ocular nos Estados Unidos

ABSTRACT Purpose: We aimed to study the characteristics of ocular trauma, an important but largely preventable global cause of blindness, in the United States. Methods: Retrospective chart review of the National Trauma Data Bank (2008-2014) was performed. All patients with ocular trauma were identified using ICD-9CM codes. The collected data were statistically analyzed with student's t-test, Chi-squared test, and logistic regression analysis performed using the SPSS software. The significance was set at p<0.05. Results: It was found that 316,485 (5.93%) of the 5,336,575 admitted trauma patients had ocular injuries. Their mean (SD) age was 41.8 (23) years, and most of them were men (69.4%). Race/ethnicity distribution was White 66.1%, Black 15.1%, and Hispanic 12.3%. The common injuries were orbital 39.5% and eye/adnexa contusions 34%. Associated traumatic brain injury was present in 58.2%. The frequent mechanisms were falls 25.5%, motor vehicle accident-occupant 21.8%, and struck by/against 17.6%. Patients <21 years of age had higher odds of cut/pierce injuries (OR=3.29, 95%CI=3.07-3.51) than the other age groups, those aged 21-64 years had higher odds of motor vehicle accident-cyclist (OR=4.95, 95%CI=4.71-5.19), and those >65 years had higher odds of falls (OR=16.75, 95%CI=16.39-17.12); p<0.001. The Blacks had a greater likelihood of firearm injuries (OR=3.24, 95%CI=3.10-3.39) than the other racial/ethnic groups, the Hispanics experienced more of cut/pierce injuries (OR=2.01, 95%CI=1.85-2.18), and the Whites experienced more of falls (OR=2.3, 95%CI=2.3-2.4); p<0.001. The Blacks (OR=3.41, 95%CI=3.34-3.48) and Hispanics (OR=1.75, 95%CI=1.71-1.79) mostly suffered assaults, while the Whites suffered unintentional injuries (OR=2.78 95%CI=2.74-2.84); p<0.001. Optic nerve/visual pathway injuries had the greatest association with very severe injury severity scores (OR=3.27, 95%CI=3.05-3.49) and severe Glasgow Coma Scores (OR=3.30, 95%CI=3.08-3.54); p<0.001. The mortality rate was 3.9%. Conclusions: Male preponderance and falls, motor vehicle accident-occupant, and struck by/against mechanisms agree with the previous reports. The identified demographic patterns underscore the need to develop group-specific preventive measures.

RESUMO Objetivo: O trauma ocular é uma causa importante e amplamente evitável de cegueira em todo o mundo. Nosso objetivo é estudar suas características nos EUA. Métodos: Revisão retrospectiva do National Trauma Data Bank (2008-2014). Todos os pacientes com trauma ocular foram identificados com códigos CID-9CM. Os dados coletados foram analisados estatísticamente e o teste t de student foi utilizado. As análises qui-quadrado e de regressão logística foram realizadas com o software SPSS. A significância foi estabelecida em p<0,05. Resultados: 316.485 (5,93%) de 5.336.575 pacientes internados com trauma, apresentaram lesões oculares. A média (DP) de idade foi de 41,8 (23) anos. A maioria era do sexo masculino (69,4%). A distribuição raça/etnia foi branca: 66,1%, negra; 15,1% e hispânica: 12,3%. As lesões comuns foram orbitárias: 39,5% e contusões dos olhos/anexos: 34%. A maioria (58,2%) teve lesão cerebral traumática. Os mecanismos frequentes foram: quedas: 25,5%, acidente com veículos motorizados: 21,8% e acidentes atingidos por algo/contra algo: 17,6%. Os pacientes <21 anos apresentaram chance aumentada de lesões com corte/perfuração (RC=3,29; IC95% = 3,07-3,51) do que outras faixas etárias, aqueles entre 21-64 anos responderam por acidente automobilístico-ciclista: (RC=4,95; IC95% = 4,71-5,19) e aqueles >65 anos foram vítimas de quedas (RC=16,75; IC 95% = 16,39-17,12); p<0,001. Os negros apresentaram maior probabilidade de lesões por arma de fogo (RC=3,24; IC95% = 3,10-3,39) do que outras raças/etnias e os hispânicos tiveram mais lesões de corte/perfuração (RC=2,01; IC95%= 1,85-2,18) enquanto os brancos tiveram mais quedas: (RC=2,3; IC95%= 2,3-2,4); p<0,001. Os negros (RC=3,41; IC95% = 3,34-3,48) e os hispânicos (RC=1,75; IC95% = 1,71-1,79) sofreram principalmente agressões e os brancos tiveram lesões não intencionais (RC=2,78; IC95% 2,74-2,84); p<0,001. Lesões do nervo óptico/via visual apresentaram maior associação com escore de gravidade de lesão muito grave (RC=3,27; IC95%= 3,05-3,49) e escores graves de Coma de Glasgow (RC=3,30; IC95%= 3,08-3,54); p<0,001. A taxa de mortalidade foi de 3,9%. Conclusões: Houve preponderância masculina a quedas, a acidentes com veículo motorizado atingidos por algo/contra algo conforme os relatos anteriores. Os padrões demográficos identificados realçam a necessidade de desenvolver medidas de prevenção específicas para os diferentes grupos.

Differential Patterns of Change in Brain Connectivity Resulting from Severe Traumatic Brain Injury.

Background: Traumatic brain injury (TBI) damages white matter tracts, disrupting brain network structure and communication. There exists a wide heterogeneity in the pattern of structural damage associated with injury, as well as a large heterogeneity in behavioral outcomes. However, little is known about the relationship between changes in network connectivity and clinical outcomes. Materials and Methods: We utilize the rat lateral fluid-percussion injury model of severe TBI to study differences in brain connectivity in 8 animals that received the insult and 11 animals that received only a craniectomy. Diffusion tensor imaging is performed 5 weeks after the injury and network theory is used to investigate changes in white matter connectivity. Results: We find that (1) global network measures are not able to distinguish between healthy and injured animals; (2) injury induced alterations predominantly exist in a subset of connections (subnetworks) distributed throughout the brain; and (3) injured animals can be divided into subgroups based on changes in network motifs-measures of local structural connectivity. In addition, alterations in predicted functional connectivity indicate that the subgroups have different propensities to synchronize brain activity, which could relate to the heterogeneity of clinical outcomes. Discussion: These results suggest that network measures can be used to quantify progressive changes in brain connectivity due to injury and differentiate among subpopulations with similar injuries, but different pathological trajectories.

Characteristics of ocular trauma in the United States.

PURPOSE: We aimed to study the characteristics of ocular trauma, an important but largely preventable global cause of blindness, in the United States. METHODS: Retrospective chart review of the National Trauma Data Bank (2008-2014) was performed. All patients with ocular trauma were identified using ICD-9CM codes. The collected data were statistically analyzed with student's t-test, Chi-squared test, and logistic regression analysis performed using the SPSS software. The significance was set at p<0.05. RESULTS: It was found that 316,485 (5.93%) of the 5,336,575 admitted trauma patients had ocular injuries. Their mean (SD) age was 41.8 (23) years, and most of them were men (69.4%). Race/ethnicity distribution was White 66.1%, Black 15.1%, and Hispanic 12.3%. The common injuries were orbital 39.5% and eye/adnexa contusions 34%. Associated traumatic brain injury was present in 58.2%. The frequent mechanisms were falls 25.5%, motor vehicle accident-occupant 21.8%, and struck by/against 17.6%. Patients <21 years of age had higher odds of cut/pierce injuries (OR=3.29, 95%CI=3.07-3.51) than the other age groups, those aged 21-64 years had higher odds of motor vehicle accident-cyclist (OR=4.95, 95%CI=4.71-5.19), and those >65 years had higher odds of falls (OR=16.75, 95%CI=16.39-17.12); p<0.001. The Blacks had a greater likelihood of firearm injuries (OR=3.24, 95%CI=3.10-3.39) than the other racial/ethnic groups, the Hispanics experienced more of cut/pierce injuries (OR=2.01, 95%CI=1.85-2.18), and the Whites experienced more of falls (OR=2.3, 95%CI=2.3-2.4); p<0.001. The Blacks (OR=3.41, 95%CI=3.34-3.48) and Hispanics (OR=1.75, 95%CI=1.71-1.79) mostly suffered assaults, while the Whites suffered unintentional injuries (OR=2.78 95%CI=2.74-2.84); p<0.001. Optic nerve/visual pathway injuries had the greatest association with very severe injury severity scores (OR=3.27, 95%CI=3.05-3.49) and severe Glasgow Coma Scores (OR=3.30, 95%CI=3.08-3.54); p<0.001. The mortality rate was 3.9%. CONCLUSIONS: Male preponderance and falls, motor vehicle accident-occupant, and struck by/against mechanisms agree with the previous reports. The identified demographic patterns underscore the need to develop group-specific preventive measures.

Sensitivity of unilateral- versus bilateral-onset spike-wave discharges to ethosuximide and carbamazepine in the fluid percussion injury rat model of traumatic brain injury.

Unilateral-onset spike-wave discharges (SWDs) following fluid percussion injury (FPI) in rats have been used for nearly two decades as a model for complex partial seizures in human posttraumatic epilepsy (PTE). This study determined if SWDs with a unilateral versus bilateral cortical onset differed. In this experiment, 2-mo-old rats received severe FPI (3 atm) or sham surgery and were instrumented for chronic video-electrocorticography (ECoG) recording (up to 9 mo). The antiseizure drug, carbamazepine (CBZ), and the antiabsence drug, ethosuximide (ETX), were administered separately to determine if they selectively suppressed unilateral- versus bilateral-onset SWDs, respectively. SWDs did not significantly differ between FPI and sham rats on any measured parameter (wave-shape, frequency spectrum, duration, or age-related progression), including unilateral (∼17%) versus bilateral (∼83%) onsets. SWDs with a unilateral onset preferentially originated ipsilateral to the craniotomy in both FPI and sham rats, suggesting that the unilateral-onset SWDs were related to surgical injury and not specifically to FPI. ETX profoundly suppressed SWDs with either unilateral or bilateral onsets, and CBZ had no effect on either type of SWD. These results suggest that SWDs with either a unilateral or bilateral onset have a pharmacosensitivity similar to absence seizures and are very different from the complex partial seizures of PTE. Therefore, SWDs with a unilateral onset after FPI are not a model of the complex partial seizures that occur in PTE, and their use for finding new treatments for PTE could be counterproductive, particularly if their close similarity to normal brain oscillations is not acknowledged.NEW & NOTEWORTHY Unilateral-onset spike-wave discharges (SWDs) in rats have been used to model complex partial seizures in human posttraumatic epilepsy (PTE), compared to bilateral-onset SWDs thought to reflect human absence seizures. Here, we show that both unilateral- and bilateral-onset SWDs following traumatic brain injury are suppressed by the antiabsence drug ethosuximide and are unaffected by the antiseizure drug carbamazepine. We propose that unilateral-onset SWDs are not useful for studying mechanisms of, or treatments for, PTE.

Potential Neuroprotective Mechanisms of Methamphetamine Treatment in Traumatic Brain Injury Defined by Large-Scale IonStar-Based Quantitative Proteomics.

Although traumatic brain injury (TBI) causes hospitalizations and mortality worldwide, there are no approved neuroprotective treatments, partly due to a poor understanding of the molecular mechanisms underlying TBI neuropathology and neuroprotection. We previously reported that the administration of low-dose methamphetamine (MA) induced significant functional/cognitive improvements following severe TBI in rats. We further demonstrated that MA mediates neuroprotection in part, via dopamine-dependent activation of the PI3K-AKT pathway. Here, we further investigated the proteomic changes within the rat cortex and hippocampus following mild TBI (TM), severe TBI (TS), or severe TBI plus MA treatment (TSm) compared to sham operated controls. We identified 402 and 801 altered proteins (APs) with high confidence in cortical and hippocampal tissues, respectively. The overall profile of APs observed in TSm rats more closely resembled those seen in TM rather than TS rats. Pathway analysis suggested beneficial roles for acute signaling through IL-6, TGFß, and IL-1ß. Moreover, changes in fibrinogen levels observed in TSm rats suggested a potential role for these proteins in reducing/preventing TBI-induced coagulopathies. These data facilitate further investigations to identify specific pathways and proteins that may serve as key targets for the development of neuroprotective therapies.

Firearm-associated ocular injuries: analysis of national trauma data.

PURPOSE: The United States of America has the highest gun ownership rate of all high-income nations, and firearms have been identified as a leading cause of ocular trauma and visual impairment. The purpose of this study was to characterize firearm-associated ocular injury and identify at-risk groups. METHODS: Patients admitted with firearm-associated ocular injury were identified from the National Trauma Data Bank (2008-2014) using the International Classification of Diseases, Ninth Revision, Clinical Modification diagnostic codes and E-codes for external causes. Statistical analysis was performed using the SPSS 24 software. Significance was set at p<0.05. RESULTS: Of the 235,254 patients, 8,715 (3.7%) admitted with firearm-associated trauma had ocular injuries. Mean (standard deviation) age was 33.8 (16.9) years. Most were males (85.7%), White (46.6%), and from the South (42.9%). Black patients comprised 35% of cases. Common injuries were orbital fractures (38.6%) and open globe injuries (34.7%). Frequent locations of injury were at home (43.8%) and on the street (21.4%). Black patients had the highest risk of experiencing assault (odds ratio [OR]: 9.0; 95% confidence interval [CI]: 8.02-10.11; p<0.001) and street location of injury (OR: 3.05; 95% CI: 2.74-3.39; p<0.001), while White patients had the highest risk of self--inflicted injury (OR: 10.53; 95% CI: 9.39-11.81; p<0.001) and home location of injury (OR: 3.64; 95% CI: 3.33-3.98; p<0.001). There was a steadily increasing risk of self-inflicted injuries with age peaking in those >80 years (OR: 12.01; 95% CI: 7.49-19.23; p<0.001). Mean (standard deviation) Glasgow Coma Scale and injury severity scores were 10 (5.5) and 18.6 (13.0), respectively. Most injuries (53.1%) were classified as severe or very severe injury, 64.6% had traumatic brain injury, and mortality occurred in 16% of cases. CONCLUSION: Most firearm-associated ocular injuries occurred in young, male, White, and Southern patients. Blacks were disproportionally affected. Most firearm-associated ocular injuries were sight--threatening and associated with traumatic brain injury. The majority survived, with potential long-term disabilities. The demographic differences identified in this study may represent potential targets for prevention.

Firearm-associated ocular injuries: analysis of national trauma data / Lesões oculares relacionadas a armas de fogo: análise de dados nacionais de traumas nos EUA

ABSTRACT Purpose: The United States of America has the highest gun ownership rate of all high-income nations, and firearms have been identified as a leading cause of ocular trauma and visual impairment. The purpose of this study was to characterize firearm-associated ocular injury and identify at-risk groups. Methods: Patients admitted with firearm-associated ocular injury were identified from the National Trauma Data Bank (2008-2014) using the International Classification of Diseases, Ninth Revision, Clinical Modification diagnostic codes and E-codes for external causes. Statistical analysis was performed using the SPSS 24 software. Significance was set at p<0.05. Results: Of the 235,254 patients, 8,715 (3.7%) admitted with firearm-associated trauma had ocular injuries. Mean (standard deviation) age was 33.8 (16.9) years. Most were males (85.7%), White (46.6%), and from the South (42.9%). Black patients comprised 35% of cases. Common injuries were orbital fractures (38.6%) and open globe injuries (34.7%). Frequent locations of injury were at home (43.8%) and on the street (21.4%). Black patients had the highest risk of experiencing assault (odds ratio [OR]: 9.0; 95% confidence interval [CI]: 8.02-10.11; p<0.001) and street location of injury (OR: 3.05; 95% CI: 2.74-3.39; p<0.001), while White patients had the highest risk of self-­inflicted injury (OR: 10.53; 95% CI: 9.39-11.81; p<0.001) and home location of injury (OR: 3.64; 95% CI: 3.33-3.98; p<0.001). There was a steadily increasing risk of self-inflicted injuries with age peaking in those >80 years (OR: 12.01; 95% CI: 7.49-19.23; p<0.001). Mean (standard deviation) Glasgow Coma Scale and injury severity scores were 10 (5.5) and 18.6 (13.0), respectively. Most injuries (53.1%) were classified as severe or very severe injury, 64.6% had traumatic brain injury, and mortality occurred in 16% of cases. Conclusion: Most firearm-associated ocular injuries occurred in young, male, White, and Southern patients. Blacks were disproportionally affected. Most firearm-associated ocular injuries were sight-­threatening and associated with traumatic brain injury. The majority survived, with potential long-term disabilities. The demographic differences identified in this study may represent potential targets for prevention.

RESUMO Objetivo: Os Estados Unidos têm a maior taxa de posse de armas de fogo de todos os países de alta renda e essas armas foram identificados como uma das maiores causas de trauma ocular e deficiência visual. O objetivo deste estudo foi caracterizar as lesões oculares associadas a armas de fogo e identificar grupos de risco. Métodos: Foram identificados pacientes hospitalizados com lesões oculares associadas a armas de fogo no período de 2008 a 2014, a partir do Banco de Dados Nacional de Trauma (National Trauma Data Bank), usando os códigos de diagnósticos da CID9MC e códigos "E" para causas externas. A análise estatística foi efetuada usando o programa SPSS. O nível de significância considerado foi de p<0,05. Resultados: De um total de 235.254 pacientes hospitalizados com trauma associado a armas de fogo, 8.715 (3,7%) tinham lesões oculares. A média de idade foi de 33,8 (DP 16,9) anos. A maioria foi de homens (85,7%), brancos (46,6%) e da região Sul (42,9%); 35% dos pacientes eram negros. As lesões mais comuns foram fraturas de órbita (38,6%) e lesões de globo aberto (34,7%). Os locais mais frequentes foram a residência (43,8%) e a rua (21,4%). Pacientes negros tiveram maior probabilidade de sofrer agressões (RP=9,0, IC 95%=8,02-10,11; p<0,001) e da ocorrência ser na rua (RP=3,05, IC 95%=2,74-3,39; p<0,001), enquanto pacientes brancos tiveram maior probabilidade de lesões autoprovocadas (RP=10,53, IC 95%=9,39-11,81; p<0,001) e da ocorrência ser na residência (RP=3,64, IC 95%=3,33-3,98; p<0,001). A probabilidade de lesões autoprovocadas aumentou com a idade de forma consistente, atingindo o máximo em pacientes com mais de 80 anos (RP=12,01, IC 95%=7,49-19,23; p<0,001). A pontuação média na escala de coma de Glasgow foi 10 (DP 5,5) e na escala de severidade da lesão foi 18,6 (DP 13,0). A maioria das lesões (53,1%) foi classificada como severa ou muito severa. Dentre os pacientes, 64,6% tiveram lesão cerebral traumática e 16% evoluíram a óbito. Conclusão: A maior parte das lesões oculares relacionadas a armas de fogo ocorreu em pacientes jovens, do sexo masculino, brancos e sulistas. Negros foram afetados desproporcionalmente. A maior parte das lesões oculares relacionadas a armas de fogo apresentou riscos à visão e foi associada a lesões cerebrais traumáticas. A maioria dos pacientes sobreviveu, mas com potencial para invalidez no longo prazo. As diferenças demográficas identificadas podem ser potencialmente alvos de ações preventivas.

Patterns of ocular trauma in elderly patients in an urban population-the Bronx experience / Padrões de trauma ocular em pacientes idosos em uma população urbana - a experiência de Bronx

ABSTRACT Purpose: To evaluate the characteristics of ocular injuries among elderly patients admitted to an urban level I trauma center because of major trauma from 2008 to 2015. Methods: A retrospective chart review was conducted of patients aged >65 years admitted with ocular injuries that were identified with ICD-9 codes. Tabulated data were analyzed using the Student's paired t-test, the chi-squared test, and regression analysis using STATA/MP-12 software. Significance was set at p<0.05. Results: Of a total of 861 patients, 221 (25.7%) admitted for major trauma and ocular injuries were elderly. The mean age of these patients was 80.3 years (median =79.2 years; interquartile range=63.8-94.6 years). Of these patients, 40.7% were males and 59.3% were females. The males were younger than the females (mean age, 77.3 vs. 82.4 years, respectively, p<0.001). Race was documented as white (30.8%), black (13.6%), and "other" (54.3%), with 67.5% of the "other" group (36.7% overall) identified as Hispanic. The most frequent injuries were contusion of the eye/adnexa (68.2%), orbital wall fractures (22.2%), and an open wound of the ocular adnexa (18.1%). Males had a 2.64-fold greater risk of orbital wall fractures (95% confidence interval [CI]=1.38-5.05, p<0.003). Patients with orbital wall fractures had higher injury severity scores than those without (95% CI=14.1-20.9 vs. 6.8-8.6, respectively, p<0.001). The most common injuries were falls (77.8%) and pedestrian/motor vehicle accidents (6.8%). Falls occurred mostly at home (51.7%), on the street (13.9%), and in hospitals/nursing homes (12.2%). Those falling at home were older than those falling at other locations (95% CI=81.8-85.4 vs. 77.0-80.6 years, respectively, p<0.002). Conclusions: Ocular injuries in elderly Bronx patients most commonly occurred in females due to falls in the home/nursing home setting. Public health measures addressing identifiable individual and environmental risks in these common locations would be most beneficial in reducing the incidence of ocular injuries in this population.

RESUMO Objetivo: Avaliar as características das lesões oculares de idosos nas internações por grandes traumatismos em um centro urbano de trauma nível I de 2008 a 2015. Métodos: Realizou-se uma revisão retrospectiva de prontuários de pacientes com mais de 65 anos internados com lesões oculares identificados com os códigos CID-9. Os dados tabulados foram analisados com o teste t de Student, teste qui-quadrado e análise de regressão, utilizando o software STATA/MP-12. A significância estatística foi fixada em p<0,05. Resultados: Duzentos e vinte e um (25,7%) pacientes de um total de 861, admitidos por traumatismo craniano importante e lesões oculares, eram idosos. A idade média era de 80,3 anos (mediana=79,2; intervalo interquartil=63,8-94,6). 40,7% eram do sexo masculino e 59,3% do feminino. Os homens eram menos idosos (média=77,3) do que as mulheres (média=82,4), p<0,001. A raça foi documentada como branca (30,8%), negra (13,6%) e "outra" (54,3%); 67,5% dos "outros" (36,7% no geral) identificados como hispânicos. As lesões mais frequentes foram contusão do olho/anexos (68,2%), fraturas da parede orbital (22,2%) e ferida aberta dos anexos oculares (18,1%). Os homens tiveram 2,64 mais chances de fraturas da parede orbital (95% CI=1,38-5,05; p<0,003). Pacientes com fraturas da parede orbital tiveram maiores escores de gravidade da lesão (95% CI=14,1-20,9) do que aqueles sem fraturas (96% IC=6,8-8,6), p<0,001. Os mecanismos comuns foram quedas (77,8%) e acidentes a pé com veículos automotores (6,8%). As quedas ocorreram principal mente em casa (51,7%), na rua (13,9%) e em hospitais/lares de idosos (12,2%). Aqueles que caíram em casa eram mais velhos (IC 95%=81,8-85,4) do que os que tiveram quedas em outros locais (IC 95%=77,0-80,6), p<0,002. Conclusões: Lesões oculares em pacientes idosos de Bronx foram mais comuns no sexo feminino e devido a quedas que ocorreram em casa/lar de idosos. Medidas de saúde pública direcionadas a riscos individuais e ambientais identificáveis nesses locais comuns seriam mais benéficas na redução de lesões oculares nessa população.

Patterns of ocular trauma in elderly patients in an urban population-the Bronx experience.

PURPOSE: To evaluate the characteristics of ocular injuries among elderly patients admitted to an urban level I trauma center because of major trauma from 2008 to 2015. METHODS: A retrospective chart review was conducted of patients aged >65 years admitted with ocular injuries that were identified with ICD-9 codes. Tabulated data were analyzed using the Student's paired t-test, the chi-squared test, and regression analysis using STATA/MP-12 software. Significance was set at p<0.05. RESULTS: Of a total of 861 patients, 221 (25.7%) admitted for major trauma and ocular injuries were elderly. The mean age of these patients was 80.3 years (median =79.2 years; interquartile range=63.8-94.6 years). Of these patients, 40.7% were males and 59.3% were females. The males were younger than the females (mean age, 77.3 vs. 82.4 years, respectively, p<0.001). Race was documented as white (30.8%), black (13.6%), and "other" (54.3%), with 67.5% of the "other" group (36.7% overall) identified as Hispanic. The most frequent injuries were contusion of the eye/adnexa (68.2%), orbital wall fractures (22.2%), and an open wound of the ocular adnexa (18.1%). Males had a 2.64-fold greater risk of orbital wall fractures (95% confidence interval [CI]=1.38-5.05, p<0.003). Patients with orbital wall fractures had higher injury severity scores than those without (95% CI=14.1-20.9 vs. 6.8-8.6, respectively, p<0.001). The most common injuries were falls (77.8%) and pedestrian/motor vehicle accidents (6.8%). Falls occurred mostly at home (51.7%), on the street (13.9%), and in hospitals/nursing homes (12.2%). Those falling at home were older than those falling at other locations (95% CI=81.8-85.4 vs. 77.0-80.6 years, respectively, p<0.002). CONCLUSIONS: Ocular injuries in elderly Bronx patients most commonly occurred in females due to falls in the home/nursing home setting. Public health measures addressing identifiable individual and environmental risks in these common locations would be most beneficial in reducing the incidence of ocular injuries in this population.

In Vitro and In Vivo Pipeline for Validation of Disease-Modifying Effects of Systems Biology-Derived Network Treatments for Traumatic Brain Injury-Lessons Learned.

We developed a pipeline for the discovery of transcriptomics-derived disease-modifying therapies and used it to validate treatments in vitro and in vivo that could be repurposed for TBI treatment. Desmethylclomipramine, ionomycin, sirolimus and trimipramine, identified by in silico LINCS analysis as candidate treatments modulating the TBI-induced transcriptomics networks, were tested in neuron-BV2 microglial co-cultures, using tumour necrosis factor α as a monitoring biomarker for neuroinflammation, nitrite for nitric oxide-mediated neurotoxicity and microtubule associated protein 2-based immunostaining for neuronal survival. Based on (a) therapeutic time window in silico, (b) blood-brain barrier penetration and water solubility, (c) anti-inflammatory and neuroprotective effects in vitro (p < 0.05) and (d) target engagement of Nrf2 target genes (p < 0.05), desmethylclomipramine was validated in a lateral fluid-percussion model of TBI in rats. Despite the favourable in silico and in vitro outcomes, in vivo assessment of clomipramine, which metabolizes to desmethylclomipramine, failed to demonstrate favourable effects on motor and memory tests. In fact, clomipramine treatment worsened the composite neuroscore (p < 0.05). Weight loss (p < 0.05) and prolonged upregulation of plasma cytokines (p < 0.05) may have contributed to the worsened somatomotor outcome. Our pipeline provides a rational stepwise procedure for evaluating favourable and unfavourable effects of systems-biology discovered compounds that modulate post-TBI transcriptomics.

Use of a Wireless Video-EEG System to Monitor Epileptiform Discharges Following Lateral Fluid-Percussion Induced Traumatic Brain Injury.

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, considerable variability has been reported for the specific parameters used in different studies that have employed this model, making it difficult to harmonize and interpret the results between laboratories. For example, variability has been reported regarding the size and location of the craniectomy, how the Luer lock hub is placed relative to the craniectomy, the atmospheric pressure applied to the dura and the duration of the pressure pulse. Each of these parameters can impact injury severity, which directly correlates with the incidence of PTE. This has been manifested as a wide range of mortality rates, righting reflex times and incidence of convulsive seizures reported. Here we provide a detailed protocol for the method we have used to help facilitate harmonization between studies. We used FPI in combination with a wireless EEG telemetry system to continuously monitor for electrographic changes and detect seizure activity.  FPI is induced by creating a 5 mm craniectomy over the left hemisphere, between the Bregma and Lambda and adjacent to the lateral ridge. A Luer lock hub is secured onto the skull over the craniectomy. This hub is connected to the FPI device, and a 20-millisecond pressure pulse is delivered directly to the intact dura through pressure tubing connected to the hub via a twist lock connector. Following recovery, rats are re-anesthetized to remove the hub. Five 0.5 mm, stainless steel EEG electrode screws are placed in contact with the dura through the skull and serve as four recording electrodes and one reference electrode. The electrode wires are collected into a pedestal connector which is secured into place with bone cement. Continuous video/EEG recordings are collected for up to 4 weeks post TBI.

Visualization of thalamic calcium influx with quantitative susceptibility mapping as a potential imaging biomarker for repeated mild traumatic brain injury.

A key event in the pathophysiology of traumatic brain injury (TBI) is the influx of substantial amounts of Ca2+ into neurons, particularly in the thalamus. Detection of this calcium influx in vivo would provide a window into the biochemical mechanisms of TBI with potentially significant clinical implications. In the present work, our central hypothesis was that the Ca2+ influx could be imaged in vivo with the relatively recent MRI technique of quantitative susceptibility mapping (QSM). Wistar rats were divided into five groups: naive controls, sham-operated experimental controls, single mild TBI, repeated mild TBI, and single severe TBI. We employed the lateral fluid percussion injury (FPI) model, which replicates clinical TBI without skull fracture, performed 9.4 Tesla MRI with a 3D multi-echo gradient-echo sequence at weeks 1 and 4 post-injury, computed susceptibility maps using V-SHARP and the QUASAR-HEIDI technique, and performed histology. Sham, experimental controls animals, and injured animals did not demonstrate calcifications at 1 week after the injury. At week 4, calcifications were found in the ipsilateral thalamus of 25-50% of animals after a single TBI and 83% of animals after repeated mild TBI. The location and appearance of calcifications on stained sections was consistent with the appearance on the in vivo susceptibility maps (correlation of volumes: r = 0.7). Our findings suggest that persistent calcium deposits represent a primary pathology of repeated injury and that FPI-QSM has the potential to become a sensitive tool for studying pathophysiology related to mild TBI in vivo.

Extracellular Vesicles as Diagnostics and Therapeutics for Structural Epilepsies.

Extracellular vesicles (EVs) are small vesicles involved in intercellular communication. Data is emerging that EVs and their cargo have potential as diagnostic biomarkers and treatments for brain diseases, including traumatic brain injury and epilepsy. Here, we summarize the current knowledge regarding changes in EV numbers and cargo in status epilepticus (SE) and traumatic brain injury (TBI), which are clinically significant etiologies for acquired epileptogenesis in animals and humans. We also review encouraging data, which suggests that EVs secreted by stem cells may serve as recovery-enhancing treatments for SE and TBI. Using Gene Set Enrichment Analysis, we show that brain EV-related transcripts are positively enriched in rodent models of epileptogenesis and epilepsy, and altered in response to anti-seizure drugs. These data suggest that EVs show promise as biomarkers, treatments and drug targets for epilepsy. In parallel to gathering conceptual knowledge, analytics platforms for the isolation and analysis of EV contents need to be further developed.

Effects of Intraborehole Flow on Purging and Sampling Long-Screened or Open Wells.

Hydraulic head differences across the screened or open interval of a well significantly influence the sampled water mixture. Sample bias can occur due to an insufficient pumping rate and/or due to native groundwater displacement by intraborehole flow (IBF). Proper understanding of the sampled water mixture is crucial for accurate interpretation of environmental tracers and groundwater chemistry data, and hence groundwater characterization. This paper uses numerical modeling to quantify sample bias caused by IBF in an un-pumped high-yield well, and the influence of pumping rate and heterogeneity on the volume of pumpage required to purge an IBF plume. The results show that (1) the pumping rate must be at least an order of magnitude greater than the IBF rate to achieve permeability-weighted yield, (2) purge volume was 2.2 to 20.6 times larger than the IBF plume volume, with the ratio depending on plume location relative to hydraulic conductivity and head distributions, and (3) after an example 1000-day un-pumped period, purging required removal of at least three orders of magnitude more water than the common practice of three to five well volumes. These results highlight the importance of knowing the borehole flow regime to identify IBF inflow and outflow zones, estimate IBF rates, and to develop a strategic sampling approach.

Circulating microRNAs as biomarkers in traumatic brain injury.

Preclinical and clinical studies can be greatly improved through the inclusion of diagnostic, prognostic, predictive or pharmacodynamics biomarkers. Circulating microRNAs (miRNAs) represent highly stable targets that respond to physiological and pathological changes. MicroRNA biomarkers can be detected by highly sensitive and absolutely quantitative methods currently available in most clinical laboratories. Here we review preclinical and clinical studies that have examined circulating miRNAs as potential diagnostic and prognostic biomarkers. We also present data that suggests pharmacodynamics biomarkers can be identified that are associated with neuroprotection in general. Although circulating miRNA can serve as useful tools, it is clear their expression profiles are highly sensitive to changing conditions and are influenced by a broad range of parameters including age, sex, body mass index, injury severity, time of collection, as well as methods of processing, purification and detection. Thus, considerable effort will be required to standardize methods and experimental design conditions before circulating miRNAs can prove useful in a heterologous injury like TBI. This article is part of the Special Issue entitled "Novel Treatments for Traumatic Brain Injury".

Convulsive seizures and EEG spikes after lateral fluid-percussion injury in the rat.

The rat lateral fluid-percussion injury (FPI) model has been used extensively to study post-traumatic epilepsy (PTE). Epidemiological studies have reported that the risk of PTE is higher after more severe injury. Adult, male Wistar rats subjected to different atmospheric pressures of injury during FPI showed great variability in injury severity when functional behavior was determined based on the Neurological Severity Score (NSS) assessment. When NSS was used to select rats with the most severe FPI-induced brain injury, 63% of rats experienced at least one convulsive seizure 2-5 weeks after FPI. This same cohort of rats (i.e., selected for severe TBI based on NSS) were significantly more susceptible to PTZ-induced seizures compared to sham controls. Video/EEG recordings from a second cohort of rats with severe FPI-induced injury (based on NSS) showed a similar incidence and frequency of spike wave discharges between rats with severe TBI and sham controls. However, the rate of isolated EEG spikes was greater in rats with severe FPI-induced injury compared to sham controls. These data suggest that convulsive seizures can be obtained in FPI-treated rats when NSS is used as an inclusion criterion to select rats with severe injury. Furthermore, although spike-wave discharges were equally prevalent in rats with severe FPI and sham controls, spontaneous spikes were more prevalent in the rats with severe FPI.

Pediatric traumatic brain injury and ocular injury.

PURPOSE: Traumatic brain injury (TBI) is a leading cause of pediatric disability and mortality. Together with sight-threatening ocular injuries, TBIs may lead to devastating consequences in developing children and complicate rehabilitation. We sought to investigate this relationship in pediatric patients admitted with major trauma. METHODS: The records of pediatric patients admitted with ocular injury and concomitant TBI were reviewed retrospectively using the National Trauma Data Bank (2008-2014). RESULTS: Of 58,765 pediatric patients admitted for trauma and also had ocular injuries, 32,173 were diagnosed with TBI. Mean patient age was 12.3 ± 7 years. Most were male (69.8%) and White (61.2%). The most frequent injuries were contusion of the eye/adnexa (39.1%) and orbital fractures (35.8%). The youngest age groups had greatest odds of falls in home locations, whereas older groups were more likely to suffer motor vehicle trauma as occupants (MVTO), struck by or against (SBA) injuries, and firearms injuries in street locations (P < 0.001). Blacks and Hispanics were most likely to suffer assault (P < 0.001) and Whites, unintentional (P < 0.001) and self-inflicted (P < 0.012) injury. Blacks were at a higher risk of firearms injury, Whites of MVTO, and Hispanics of motor vehicles as pedestrians (P < 0.001). CONCLUSIONS: TBI frequently is experienced by trauma patients with concomitant ocular injury and should be considered in children admitted with major trauma. Resultant demographic patterns may help identify patients that have a higher risk of TBI leading to earlier diagnosis and treatment.

Thymosin beta 4 induces significant changes in the plasma miRNA profile following severe traumatic brain injury in the rat lateral fluid percussion injury model.

OBJECTIVES: Thymosin beta 4 (Tß4) has demonstrated neuroprotective potential in models of neurlogical injury. The neuroprotective potential of Tß4 has been associated with increased miR-200a and miR-200b within the brain following stroke. Here we tested the hypothesis that Tß4 treatment could also alter miRNA profiles within the plasma following severe traumatic brain injury (TBI). METHODS: We used the rat lateral fluid percusion injury model of severe TBI to test this hypothesis. Highly sensitive and quantitative droplet digital polymerase chain reaction (ddPCR) was used to measure the plasma concentrations of miR-200 family members. In addition, we conducted RNAseq analysis of plasma miRNA to further identify changes associated with TBI and treatment with Tß4. RESULTS: ddPCR demonstrated that miR-200a-3p andmiR-200b-3p were both significantly increased in plasma following treatment with Tß4 after severe TBI. RNAseq analysis suggested that miR-300-3p and miR-598-3p increased while miR-450-3p and miR-194-5p significantly decreased following TBI. In contrast, miR-194-5p significantly increased in Tß4 treated rats following TBI. In addition, we identified nine plasma miRNAs whose expression significantly changed following treatment with Tß4. CONCLUSIONS: Tß4 treatment significantly increased plasma levels of miR-200a-3p and miR-200b-3p, while RNAseq analysis identified miR-194-5p as a candidate miRNA that may be critical for neuroprotection.