Currently, there is no definitive diagnostic test for traumatic brain injury (TBI) to help physicians determine the seriousness of injury or the extent of cellular pathology. Calpain cleaves alphaII-spectrin into breakdown products (SBDP) after TBI and ischemia. Mean levels of both ipsilateral cortex (IC) and cerebral spinal fluid (CSF) SBDP at 2, 6, and 24 h after two levels of controlled cortical impact (1.0 mm and 1.6 mm of cortical deformation) in rats were significantly elevated by injury. CSF and IC SBDP levels were significantly higher after severe (1.6 mm) injury than mild (1.0 mm) injury over time. The correlation between CSF SBDP levels and lesion size from T2-weighted magnetic resonance images 24 hours after TBI as well as correlation of tau and S100beta was assessed. Mean levels of CSF SBDP (r = 0.833) and tau (r = 0.693) significantly correlated with lesion size while levels of CSF S100beta did not (r = 0.188). Although levels of CSF and IC SBDP and lesion size are all significantly higher after 1.6 mm than 1.0 mm injury, the correlation between CSF SBDP and lesion size was not significant following the removal of controls from the analysis. This indicates CSF SBDP is a reliable marker of the presence or absence of injury. Furthermore, larger lesion sizes 24 h after TBI were negatively correlated with motor performance on days 1-5 after TBI (r = -0.708). Based on these data, evaluation of CSF SBDP levels as a biomarker of TBI is warranted in clinical studies.

Primary septo-hippocampal cell cultures were incubated in varying concentrations of tumor necrosis factor (TNF-alpha; 0.3-500 ng/ml) to examine proteolysis of the cytoskeletal protein alpha-spectrin (240 kDa) to a signature 145 kDa fragment by calpain and to the apoptotic-linked 120-kDa fragment by caspase-3. The effects of TNF-alpha incubation on morphology and cell viability were assayed by fluorescein diacetate-propidium iodide (FDA-PI) staining, assays of lactate dehydrogenase (LDH) release, nuclear chromatin alterations (Hoechst 33258), and internucleosomal DNA fragmentation. Incubation with varying concentrations of TNF-alpha produced rapid increases in LDH release and nuclear PI uptake that were sustained over 48 hr. Incubation with 30 ng/ml TNF-alpha yielded maximal, 3-fold, increase in LDH release and was associated with caspase-specific 120-kDa fragment but not calpain-specific 145-kDa fragment as early as 3.5 hr after injury. Incubation with the pan-caspase inhibitor, carbobenzosy- Asp-CH(2)-OC (O)-2-6-dichlorobenzene (Z-D-DCB, 50-140 microM) significantly reduced LDH release produced by TNF-alpha. Apoptotic-associated oligonucleosomal-sized DNA fragmentation on agarose gels was detected from 6 to 72 hr after exposure to TNF-alpha. Histochemical changes included chromatin condensation, nuclear fragmentation, and formation of apoptotic bodies. Results of this study suggest TNF-alpha may induce caspase-3 activation but not calpain activation in septo-hippocampal cultures and that this activation of caspase-3 at least partially contributes to TNF-alpha-induced apoptosis.

Summary Five healthy Equidae (4 horses and one pony) were given a single i.v. dose of ceftriaxone (50 mg/kg bwt) to determine the pharmacokinetics and concentration in cerebrospinal fluid (CSF). Blood was drawn from an i.v. jugular catheter and CSF from a pre‐placed, intrathecal catheter. Serum and CSF concentrations were determined by high performance liquid chromatography. The mean serum concentration of ceftriaxone was 144.7 μg/ml 15 min after injection and declined to 0.3 μg/ml 10 h after injection. The elimination rate constant (Λ2) was 0.63 ± s.e. 0.23/h, the elimination half‐life (t1/2) was 1.62 ± s.e. 0.42 h apparent volume of distribution at steady state (Vd(ss)) was 330.8 ± 11.8 ml/kg bwt. Clearance was 312.7 ± 38 ml/h/kg bwt and mean residence time was 1.13 ± 0.14 h. Mean CSF concentration was 0.60 ± 0.14 μg/ml at 3 h after injection and 0.4 ± 0.31 μg/ml at 8 h. Ceftriaxone may be useful in the treatment of bacterial infections in horses. Its ability to penetrate the CSF should make it effective in the treatment of bacterial meningitis.