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Neuropathol Appl Neurobiol 2:323C332. [Google Scholar] 3. short\term cases post TBI, virtually no A plaques were found in long\term survivors. A potential mechanism for A plaque regression was suggested by the post\injury accumulation of an A degrading enzyme, neprilysin. These findings fail to support the premise that progressive plaque pathology after TBI ultimately results in AD. (1989); ICH?=?intracerebral hematoma; EDH?=?Extradural Hematoma; (e)?=?evacuated; RICP?=?raised intracranial pressure;?+?=?present;???=?absent; NK?=?not known; N/A?=?not applicable. All brain sections that were examined contained white matter tracts from the subcortex, deep white matter and corpus callosum. Use of tissue for this study was approved by the Ethics Committee of the Southern General Hospital, South Glasgow University Hospitals NHS Trust. Immunohistochemistry IHC staining was performed on serial paraffin\embedded sections. Single, double or triple labeled IHC was performed using a well\characterized panel of primary antibodies (Table?2). Table 2 Summary of antibodies used for immunohistochemistry. Abbreviations: M?=?monoclonal; P?=?polyclonal; APP?=?amyloid precursor protein; A?=?amyloid beta; NF?=?neurofilament. (A) Axonal bulb immunoreactivity for amyloid (A) (6F/3D) antibody, \site APP Palmitic acid cleaving enzyme (BACE), presenilin\1 (PS\1) and neprilysin in long\term survival cases. (B) A plaques found in short\term survival cases (4G8 antibody). Scale bar?=?50?m. Open in a separate window Figure 3 Top panels: representative triple labeling of amyloid precursor protein (APP), amyloid (A), \site APP cleaving enzyme (BACE), presenilin\1 (PS\1), kinesin, neprilysin (NEP), and APP within both short\ and long\term survival cases. Immunoreactivity gives the typical appearance of these proteins accumulating within axonal bulbs in the subcortical white matter. 13335 and Amy33 antibodies Palmitic acid demonstrate A within axonal bulbs. Neprilysin can be found co\localizing with APP and A. Scale bar?=?50?m. Bottom Panels: Immunofluorescence demonstrated macrophages/ microglia in the brain tissue in short\ and long\term survival cases detected by antibodies OX42 and macrophage inflammatory protein. Luxol fast blue and immunostaining for myelin basic protein (SMI 94) revealed patchy loss of immunoreactivity in the subcortical white matter in both short\ and long\term survival patients. Scale bar?=?50?m. Immunoreactivity for PS\1 protein in swollen axonal profiles was detected using the specific antibody PS\1. PS\1 immunoreactivity was less prevalent than that of BACE with predominantly minimal immunoreactivity being detected in both groups. Palmitic acid However, it was clear that PS\1 immunoreactivity was elevated in cases surviving up to 3 years post TBI (?(2,2, ?,33). Immunoreactivity for kinesin protein in swollen axonal profiles was identified by antibody specific to kinesin (L1). Kinesin immunoreactivity was demonstrated in almost all cases in axonal bulbs. In terms of the extent of immunoreactivity, both the long\ and short\term survival groups had moderate to extensive staining (Figure?3). Using double or triple\fluorescence IHC, we consistently found co\immunoreactivity of combinations of APP or A with BACE and PS\1 in axonal profiles by using specific antibodies (Figure?3). This co\immunoreactivity was seen in cases who died shortly after trauma as well as in those who survived long\term (Figure?3). A accumulation In total, a panel of five antibodies were used to detect various A epitopes. Initially, for detection of plaque\like profiles, 13335 was used in all sections from all cases. 4G8 Palmitic acid was used on 11 sections from 11 cases to confirm the presence or absence of plaque pathology as identified by 13335. Staining for A plaque pathology also revealed the presence of A within axon bulbs. Thus, confirmatory detection of A accumulation in damaged axons was accomplished using BC05, 6F/3D and Amy33; all of which are specific for A and do not cross\react with full\length APP. BC05 was used in all cases, 6F/3D in 16 cases, and Amy33 in 11 cases. When compared, these antibodies revealed consistent immunoreactivity to A. Axonal swellings stained for A had the typical appearance of axonal bulbs at the terminal ends of disconnected axons following TBI. Double immunostaining demonstrated co\localization of A with markers of axonal pathology (APP and NF) within some of the axonal bulbs. Although APP and A co\localization could be seen, this was not the case in all axon bulbs, with some bulbs staining with APP alone. Classically observed varicose swellings were demonstrated within non\disconnected axons, yet A ENOX1 accumulation was not detected within these sites despite the accumulation of other proteins including APP and NF. A accumulation appeared to be restricted to the axonal bulbs. Notably,.