Although accumulating evidence suggests that repetitive mild TBI (rmTBI) may cause long-term cognitive dysfunction in adults, whether rmTBI causes similar deficits in the immature brain is unknown. Here we used an experimental model of rmTBI in the immature brain to answer this question. Post-natal day (PND) 18 rats were subjected to either one, two, or three mild TBIs (mTBI) or an equivalent number of sham insults 24 h apart. After one or two mTBIs or sham insults, histology was evaluated at 7 days. After three mTBIs or sham insults, motor (d1-5), cognitive (d11-92), and histological (d21-92) outcome was evaluated. At 7 days, silver degeneration staining revealed axonal argyrophilia in the external capsule and corpus callosum after a single mTBI, with a second impact increasing axonal injury. Iba-1 immunohistochemistry showed amoeboid shaped microglia within the amygdalae bilaterally after mTBI. After three mTBI, there were no differences in beam balance, Morris water maze, and elevated plus maze performance versus sham. The rmTBI rats, however, showed impairment in novel object recognition and fear conditioning. Axonal silver staining was observed only in the external capsule on d21. Iba-1 staining did not reveal activated microglia on d21 or d92. In conclusion, mTBI results in traumatic axonal injury and microglial activation in the immature brain with repeated impact exacerbating axonal injury. The rmTBI in the immature brain leads to long-term associative learning deficit in adulthood. Defining the mechanisms damage from rmTBI in the developing brain could be vital for identification of therapies for children.

It is believed that biosynthesis of lipid mediators in the central nervous system after cerebral ischemia-reperfusion starts with phospholipid hydrolysis by calcium-dependent phospholipases and is followed by oxygenation of released fatty acids (FAs). Here, we report an alternative pathway whereby cereberal ischemia-reperfusion triggered oxygenation of a mitochondria-specific phospholipid, cardiolipin (CL), is followed by its hydrolysis to yield monolyso-CLs and oxygenated derivatives of fatty (linoleic) acids. We used a model of global cerebral ischemia-reperfusion characterized by 9 minutes of asphyxia leading to asystole followed by cardiopulmonary resuscitation in postnatal day 17 rats. Global ischemia and cardiopulmonary resuscitation resulted in: (1) selective oxidation and hydrolysis of CLs, (2) accumulation of lyso-CLs and oxygenated free FAs, (3) activation of caspase 3/7 in the brain, and (4) motor and cognitive dysfunction. On the basis of these findings, we used a mitochondria targeted nitroxide electron scavenger, which prevented CL oxidation and subsequent hydrolysis, attenuated caspase activation, and improved neurocognitive outcome when administered after cardiac arrest. These data show that calcium-independent CL oxidation and subsequent hydrolysis represent a previously unidentified pathogenic mechanism of brain injury incurred by ischemia-reperfusion and a clinically relevant therapeutic target.

Traumatic brain injury (TBI) is a major health problem associated with significant morbidity and mortality. The pathophysiology of TBI is complex involving signaling through multiple cascades, including lipid peroxidation. Oxidized free fatty acids, a prominent product of lipid peroxidation, are potent cellular mediators involved in induction and resolution of inflammation and modulation of vasomotor tone. While previous studies have assessed lipid peroxidation after TBI, to our knowledge no studies have used a systematic approach to quantify the global oxidative changes in free fatty acids. In this study, we identified and quantified 244 free fatty acid oxidation products using a newly developed global liquid chromatography tandem-mass spectrometry (LC-MS/MS) method. This methodology was used to follow the time course of these lipid species in the contusional cortex of our pediatric rat model of TBI. We show that oxidation peaked at 1 hr after controlled cortical impact and was progressively attenuated at 4 and 24 hr time points. While enzymatic and non-enzymatic pathways were activated at 1 hr post-TBI, enzymatic lipid peroxidation was the predominant mechanism with 15-lipoxygenase (LOX) contributing to the majority of total oxidized fatty acid content. Pro-inflammatory lipid mediators were significantly increased at 1 and 4 hr after TBI with return to basal levels by 24 hr. Anti-inflammatory lipid mediators remained significantly increased across all three time points, indicating an elevated and sustained anti-inflammatory response following TBI.

Fifty-four individually-fed Hereford-Angus cross steers (initial BW = 308 ± 9 kg) were used in an unbalanced randomized block design with a 2 × 2 factorial treatment arrangement to determine effects of corn processing method and corn wet distillers grains plus solubles (WDGS) inclusion in finishing diets on animal performance, carcass and beef characteristics, and sensory attributes. Dietary treatments included steam-flaked corn- (SFC) and dry-rolled corn (DRC)-based finishing diets containing 0 or 35% WDGS (DM basis; 0SFC and 35SFC, 0DRC and 35DRC, respectively). Yellow grease was used to equilibrate fat content of diets. Steers were fed 174 d, and were harvested on a single date when the mean ultrasound fat thickness was estimated to be 1.30 cm. No interactions between corn processing and WDGS were observed for performance or carcass characteristics (P ≥ 0.11). Final BW (556 ± 14 kg) and ADG (1.43 ± 0.06 kg) were not affected (P ≥ 0.25) by dietary treatment. Steers fed SFC-based diets consumed less feed, and were 10.6% more efficient (P < 0.01) than those fed DRC-based diets. Including WDGS in finishing diets improved feed efficiency of steers consuming both SFC- and DRC-based diets (P ≤ 0.04). Dietary treatment did not affect HCW, dressing percentage, fat thickness, or yield grade (P ≥ 0.27). Including WDGS in finishing diets decreased the concentration of 16:1cis-9, 18:1cis-9, and 18:1cis-11 fatty acids, and tended (P ≤ 0.10) to increase total fat concentration of steaks compared with diets without WDGS. A corn processing method by WDGS interaction was detected for 18:1trans-11 where steaks from 0DRC diets had decreased concentrations compared with other diets. There were no dietary effects on palatability attributes (P > 0.20). Livery-organy aromatics (P = 0.03) and sweet basic tastes (P = 0.01) in steaks from the 35SFC treatment were more intense than in other treatments, but were barely detectable. Thiobarbituric acid reactive substances tended to be greater in steaks from steers fed WDGS after 5 d of storage (P = 0.10), and were greater after 7 d. (P < 0.01). Inclusion of WDGS used in this experiment improved G:F with minimal impacts on carcass characteristics. Both WDGS inclusion and corn processing method impacted fatty acid composition. However, diet had minimal impacts on palatability attributes. When compared with diets fat-equilibrated with yellow grease, the primary concern with incorporating WDGS appears to be decreased shelf-life after 5 d of storage.

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