Background - We have recently developed and characterized a rat model of mild traumatic brain injury which simulates the concussive injuries frequently encountered by players in American professional football. Objectives - To study the effect of multiple impacts to the head on intracranial pressure, cognitive function, and exploratory behavior. Materials and methods - The model was employed to cause concussion. Intracranial pressure, cognitive function, and exploratory behavior were examined following the multiple impacts of a 50 or 100 g projectile at a velocity of 9.3 or 11.2 m/s to the helmet protected head. Results - Intracranial pressure measured at 6 and 10 h, and 1, 2, 3, 5, and 7 days. It was maximally elevated 10 h after impact and returned to the control levels 7 days later. Morris Water Maze assessment, 48 h after impact, revealed impaired cognitive function. Open field testing 2-4 days and 1 and 2 weeks after impacts indicated consistently reduced spontaneous exploratory activity. Conclusion - Multiple impacts to the head raise intracranial pressure and impair cognitive function and exploratory activity in this animal model.

Three animal models were evaluated in this study involving head impacts of the rat, including the Marmarou drop-weight and two momentum-exchange techniques. In series 1, 36 Wistar rats were hit on the side of the free-moving head using Marmarou's 450 g impact mass at 4.4, 5.4, and 6.3 m/s. Head acceleration was measured and injuries were observed. The 6.3-m/s side impact resulted in no deaths, no skull fractures, infrequent contusions, and some injuries consistent with diffuse axonal injury. In series 2, 57 Marmarou drop-weight tests were conducted to study head biomechanical responses. Marmarou's technique involves a head impact followed by prolonged loading into a foam pad under the animal. Based on the literature, the 2 m (6.3 m/s) Marmarou drop causes death, skull fracture, brain and spinal cord contusions, and diffuse axonal injury. These injuries are more severe than that occurring with impact of similar mass and velocity to the free-moving head. Impacts to the free-moving head provide more realistic animal models to study concussion and severe brain injury.

There is increasing evidence that even low levels of blast cause brain injury, but little is known about their thresholds and mechanisms. Exposure of rats to 10-60 kPa blasts elevate intracranial pressure (ICP) in a dose-dependent manner and impair cognitive function. We have evaluated a prophylactic measure against these brain injuries in a rat animal model, consisting of feeding them processed cereal. This type of feed is known to ameliorate disturbances in secretion of body fluids and to have anti-inflammatory effects. In humans, intake of processed cereals is effective against intestinal diarrhea and also reduces the symptoms of Ménière's disease. Rats were given either standard laboratory feed or processed cereal feed for 2 weeks before exposure to blast in a shock tube. The ICP was monitored at different time points up to 1 week after exposure to a 60-kPa blast, and for up to 24 h after exposure to a 30-kPa blast. Maximal ICP elevation was reached at 10 h in both groups. In the group of rats on standard feed exposed to 60 kPa, an ICP increase of 145% was noted at 10 h, and the corresponding increase in the rats fed processed cereal feed was only 50%. In rats exposed to a 30-kPa blast, those fed standard feed and processed cereal feed demonstrated increases of ICP of 80% and 40%, respectively. Cognitive function as measured by the Morris water maze was assessed in other groups of rats at 2 days after exposure to 10- or 30-kPa blasts. Their performance was significantly impaired at both exposure levels in rats on standard feed, but no functional impairment was seen in rats fed processed cereal feed.

The symptoms of primary blast-induced mTBI, posttraumatic stress disorder and depression overlap. Evidence of an organic basis for these entities has been scarce and controversial. We present a review of animal studies demonstrating that low-level blast causes pathophysiological and functional changes in the brain. We monitor a time period from minutes to approximately 1week after blast exposure from multiple modes (air, underwater, localized and whole body). The most salient findings observed were (1) the peak pressures (Pmax) in the brain, elicited from the blast from the firing of military weapons (Pmax 23–45kPa), have a similar magnitude as that registered in air close to the head. Corresponding measurements during the detonation pulse from explosives under water show a Pmax in the brain, which is only 10% of that in water outside the head. (2) The rise time of the pressure curve is 10 times longer in the brain as compared with the blast in air outside the head during firing of military weapons. (3) The lower frequencies in the blast wave appear to be transmitted more readily to the brain than the higher frequencies. (4) When animals are exposed to low levels of blast, the blast wave appears mostly transmitted directly to the brain during air exposure, not via the thorax or abdomen. (5) Low levels of blast cause brain edema, as indicated by increased bioelectrical impedance, an increase in the intracranial pressure, small brain hemorrhages and impaired cognitive function.

Blast injury is the leading cause of traumatic brain injury (TBI) in military personnel serving in Iraq and Afghanistan. TBI, caused by blast exposure, has considerable economic, military and social implications. Blast injuries are frequently caused by the high amplitude blasts generated by improvised explosive devises (IEDs). However, even regular training with large caliber weapons appears to cause mild traumatic brain injury (mTBI). The occupational standards of the Armed Forces provide guidelines regarding the maximal peak pressures of blasts and the number of exposures allowed per day, week, etc. These standards are based on acute injuries in ears and/or lungs and not on short- or long term effects on the brain.This paper investigates numerically the propagation of blast waves from the AG90 rifle and through the skull bone and into the brain, in order to understand better of how blast waves can cause brain injury. The numerical simulations were performed with the hydrocode AUTODYN. An idealized head was used in the gunner's position during firing of the relevant weapon. The paper focuses on qualitative mechanisms of blast wave transmission into the brain and investigates in particular the effect of geometry and material parameters.

Anaesthetized swine in crew positions were exposed to weapons in air or to explosives underwater. Blast parameters were correlated with those in the brain. The peak pressure in the brain (Pmax brain/air) was 0.7 for a bazooka (45 kPa), 0.5 for a howitzer (10 kPa), and 0.4 for a rifle (23 kPa). The brain/water Pmax for the detonation pulse of under water explosives was only 0.1, but 0.3–0.4 for the secondary pulses. The results indicate that low-frequency spectra penetrate easier into the brain. Histological examination revealed small hemorrhages in rear regions of the brain. In rats, we investigated the effect of shock tube blasts. After exposure to 10 or 30 kPa, cognitive performance (Morris Water Maze) decreased by 50%. The intracranial pressure (ICP) increased in a dose dependent fashion to reach peak levels 6 h after exposure at 10 kPa and 10 h after exposure to 30 or 60 kPa. An initial ICP elevation took place 30 min after exposure to 60 kPa, and 2 and 6 h after exposure to 30 and 10 kPa, respectively. A prophylaxis, consisting of a 2 week intake of hydrothermally fermented cereals, reduced significantly the blast effect both on ICP and cognitive performance. [The authors thanks Svante Hjer, Samba Sensors AB. The study was supported by the Swedish Armed Forces and FMV.]

Brain injury after high-level blast has been established both clinically and experimentally. Less is known about the effects on the brain of exposure to low to moderate blast levels, such as those encountered by military personnel during the firing of weapons. This study investigates if exposure to occupational levels of low-level blasts affect intracranial pressure and cognitive performance. Rats were exposed to blast overpressure in a shock tube at peak levels of 10, 30, and 60 kPa. Intracranial pressure (ICP) was measured after 0.5, 3, 6, and 10 h and 1, 2, 3, 5, and 7 days. We found two features of the response: a dose-dependent rise in ICP in rats exposed to blast, and an increasing time delay in elevation with decreasing intensity of exposure. The ICP increased in a dose-dependent fashion, up to 15.7mm Hg after exposure to a 60-kPa blast from a control level of 6 mm Hg. While the initial elevation took place within 30 min after exposure to 60 kPa, it did not appear until after 2 and 6 h for 30 and 10 kPa, respectively. In all cases, the ICP returned to control levels after 7 days. The cognitive function of the blast-exposed rats was assessed with the Morris water maze. After exposure to 10 or 30 kPa and re-testing 2 days later, the latency was increased by over 100%. The results show that exposure of rats to blast levels as low as 10 kPa affects both ICP and cognitive function. Though species differences do not allow direct extrapolation to humans, these findings do pose the question as to whether the thresholds for brain injury might be lower than those of other organs used to set training standards for blast exposure.

Military personnel are exposed to occupational levels of blast overpressure during training. This study characterizes the pressure-time histories of air, underwater, and localized blast, and correlates blast parameters with neuropathology. Blast overpressure was produced by a howitzer, a bazooka, an automatic rifle, underwater explosives, or a shock tube. Anesthetized pigs were exposed in positions that simulated real training scenarios. Underwater exposures were performed using explosives at distances recommended by safety requirements. In other experiments, rats were exposed via a shock tube. The pressure changes were recorded with a hydrophone sensor in the brain of the pig and in rats with an optical fiber sensor. Histological examination of porcine brains revealed small parenchymal and subarachnoid hemorrhages, predominately in the occipital lobe, cerebellum, and medulla oblongata. Relative to the peak pressure in air, that in porcine brain (Pmax brain/air) was 0.7 for the bazooka and 0.5 and 0.7, respectively, for the 9- and 30-kPa howitzer. The attenuation was stronger in water: the detonation pulse had a brain/water ratio of 0.1, and the secondary pulses had ratios of 0.3-0.4. The results indicate that low-frequency spectra penetrate easily from air or water into the brain, but high-frequency spectra appear to be filtered by body structures. In addition, blast waves were recorded in the brain and abdomen of pigs after local exposure via shock tube to either the abdomen or the top of the skull. When the abdomen was exposed, the maximal peak value in the brain was only 3% of that in the abdomen. Moreover, part of this pressure could have been derived from the air outside the head. The results gave little support to significant transmission of pressure within the body.

Rotational acceleration of the head, as occurs in falls, car crashes, and sport injuries, may result in diffuse brain damage, with acute and chronic neurological and psychiatric symptoms. The present study addresses the effects of rotational trauma on the neuronal cytoskeleton, which stabilizes perikaryal, dendritic and axonal shape and function. The study focuses upon the distribution of (1) the phosphorylated form of the heavy neurofilament subunit, (2) the light neurofilament subunit, and (3) beta-amyloid, a marker for brain injury. While normally restricted to axons, the phosphorylated heavy neurofilament subunits were drastically decreased in the axons after rotational trauma. Instead, they accumulated in the neuronal perikarya, normally devoid of the phosphorylated subunit. This alteration was seen, not only in the cerebral cortex, but also in the hippocampus, the cervical spinal cord, the cerebellum, the cranial nerves and the pyramidal tract. The distribution of the light subunit of neurofilaments was also altered post trauma. Only a weak beta-amyloid immunoreactivity was detected in the brains of control animals. Promptly after the trauma, a large number of beta-amyloid positive neurons appeared. Intensely co-localized immunoreactivity for the light subunit of neurofilaments and of beta-amyloid was seen 3 days after the rotational trauma axons of in the subcortical white matter and in the granule cell layer of the dentate gyrus as well as in neurons of the hypoglossal nucleus. The reported alterations in the central nervous system neurons are similar to those in the human brain after closed head injury and in chronic degenerative diseases. Regions of importance for social behavior, memory and body movement were affected.

The aim of this study was to isolate a compound from blood plasma that inhibits intestinal diarrhea and that appears also to regulate fluid volumes in other organs. The isolation procedure included lipid extraction, liquid chromatography, and gas chromatography. The active substance was identified by mass spectrometry as erucamide (MW 337 Da). The biological effect was reproduced with authentic erucamide. Erucamide is a fatty acid amide, such as oleamide and anandamide, which modulate other physiological functions in a receptor-mediated fashion. All the exact biological functions of erucamide are as yet to be defined, but it is already known to stimulate angiogenesis. Erucamide concentrations were determined in body organs from the pig. The blood plasma level was 3 ng/g, and those of lung, kidney, liver, and brain were 12, 2.5, 1.0, and 0.5 ng/g, respectively. Erucamide was below detection level in the intestine, but is known to be present in the cerebrospinal fluid. In the rat, 3H-erucamide was accumulated in vivo into lung, liver, and spleen and in vitro into lung, liver, brain, and intestine. The in vitro uptake was time and temperature dependent, but not saturable.

The interstitial fluid of the human myocardium was monitored in 13 patients undergoing aortic valve and/or bypass surgery before, during, and after hypothermic potassium cardioplegia. The regulation of glucose and lactate was studied after sampling with microdialysis. The following questions were addressed. 1). Is the rate of transcapillary diffusion the limiting step for myocardial uptake of glucose before or after cardioplegia? 2). Does cold potassium cardioplegia induce a critical deprivation of glucose and/or accumulation of lactate in the myocardium? Before cardioplegia, interstitial glucose was approximately 50% of the plasma level (P < 0.001). Interstitial glucose decreased significantly immediately after induction of cardioplegia and remained low (1.25 +/- 0.25 mM) throughout cardioplegia. It was restored to precardioplegic levels 1 h after release of the aortic clamp. Interstitial glucose then decreased again at 25 and 35 h postoperatively to the levels observed during cardioplegia. Interstitial lactate decreased immediately after induction of cardioplegia but returned to basal level during the clamping period. At 25 and 35 h, interstitial lactate was significantly lower than before and during cardioplegia. Glucose transport over the capillary endothelium is considered rate limiting for its uptake in the working heart but not during cold potassium cardioplegia despite the glucose deprivation following perfusion of glucose-free cardioplegic solution. Lactate accumulated during cardioplegia but never reached exceedingly high interstitial levels. We conclude that microdialysis provides information that may be relevant for myocardial protection during open-heart surgery.

Exposure to impulse noise, above a certain intensity, is harmful to auditory function. Effects of impulse noise on the central nervous system (CNS) are largely unexplored, and there is little information on critical threshold values and time factors. We have recently shown that neurofilament proteins are affected in the cerebral cortex and the hippocampus. Now we show that impulse noise induces expression of the immediate early gene c-Jun products, proposed to play a role in the initiation of neuronal death, and apoptosis as revealed by TUNEL staining. Rat brains were investigated immunohistochemically 2 h to 21 days after exposure to impulse noise of 198 dB or 202 dB. c-Jun was expressed in neuronal perikarya in layers II-VI of the temporal cortex, the cingulate and the piriform cortices at 2 h to 21 days after both exposure levels. Granule neurons of the dentate gyrus and the CA1-3 in the hippocampus pyramidal neurons were similarly affected. The elevated expression of c-Jun products remained high at all postexposure times. TUNEL staining was positive among the same nerve cell populations 6 h after exposure and persisted even at 7 days at both exposure levels.

There is increasing evidence that impulse noise causes brain damage, but little is known about the mechanisms and extent of the response. Here, rat brains were investigated immunohistochemically for the expression of c-Fos, c-Myc, and beta-APP during the first 3 weeks postexposure to impulse noise of 198 or 202 dB. The expression of c-Fos and c-Myc increased at 2 h after exposure in neurons of the cerebral cortex, thalamus, and hippocampus, and this c-Fos immunoreactivity remained elevated for the entire observation period. The c-Myc immunoreactivity peaked at 18 h in both neurons and astrocytes but returned to control levels at 7 days. Abnormal deposition of beta-APP was evident within 6 h in the same brain regions. The beta-APP immunoreactivity was most prominent at 18 h and remained increased over the 21-day period assessed. The observed effects were similar to those described in humans following traumatic brain injury and in Alzheimer's disease. We conclude that impulse noise influences the brain in a fashion similar to that in cases with progressive CNS degeneration.

Objective: To investigate the time course of troponin-T release into the extracellular fluid of the myocardium and to distinguish between a rise in troponin-T due to implantation trauma and an increase due to cardiac arrest during coronary surgery. Design: Microdialysis probes were implanted in the heart of seven patients soon after sternotomy. Troponin-T was measured in the microdialysates and in peripheral blood from 3 h before to 24 h after heart arrest. Results: The troponin-T concentration in the microdialysates increased immediately after probe implantation and decreased to baseline within 70 min. This early peak is interpreted to reflect a local trauma. Three hours after crossclamp release, a second peak of microdialysate troponin-T was recorded; 50 times higher than in serum. Eight to 24 h later a third peak occurred in five patients. Serum troponin-T was below the detection level at the beginning of the operation but increased linearly during the first 3 h of reperfusion and remained at that level thereafter. Conclusion: Microdialysis is a safe technique providing more information on myocardial metabolism during and after bypass surgery than can be obtained from peripheral blood. The release of troponin-T in response to cardiac arrest can be distinguished in time from the local tissue response to probe implantation.

Exposure to impulse noise, i.e. pressure waves, is above a certain intensity, harmful to auditory function. Intense, short-lasting impulse noise of 198 or 202 dB affects the heavy subunit of neurofilament proteins in neuronal perikarya of the cerebral cortex and hippocampus. There was as well an increased expression of immediate early gene products and induction of neuronal apoptosis. Here, we show that this range of exposure also affects glial cells. We identified microglial cells with an antibody against the complement receptor type 3 (OX-42) and astrocytes with an antibody against the glial fibrillary acidic protein (GFAP). The pattern of damage included microglial activation as early as 2 h after exposure to 202 dB. The activation increased further at 18 h. There was a significant increase of the area occupied by microglial cells in the anterior and posterior hypothalamus and in the lateral septal nucleus. Astrogliosis was observed in the cerebral cortex, the dentate gyrus and in the pyramidal cell layers as well as in white matter of the hippocampus. Both the microglial and astrocytic reactivities remained at 21 days. Exposure to 198 dB, caused similar, but less prominent activation in both cell types.

Rapid head rotation is a major cause of brain damage in automobile crashes and falls. This report details a new model for rotational acceleration about the center of mass of the rabbit head. This allows the study of brain injury without translational acceleration of the head. Impact from a pneumatic cylinder was transferred to the skull surface to cause a half-sine peak acceleration of 2.1 x 10(5) rad/s2 and 0.96-ms pulse duration. Extensive subarachnoid hemorrhages and small focal bleedings were observed in the brain tissue. A pronounced reactive astrogliosis was found 8-14 days after trauma, both as networks around the focal hemorrhages and more diffusely in several brain regions. Astrocytosis was prominent in the gray matter of the cerebral cortex, layers II-V, and in the granule cell layer and around the axons of the pyramidal neurons in the hippocampus. The nuclei of cranial nerves, such as the hypoglossal and facial nerves, also showed intense astrocytosis. The new model allows study of brain injuries from head rotation in the absence of translational influences.

The aim of this study is to monitor excitatory amino acids (EAAs) in the extracellular fluids of the brain and to characterize regional neuronal damage in a new experimental model for brain injury, in which rabbits were exposed to 180-260 krad/s2 rotational head acceleration. This loading causes extensive subarachnoid hemorrhage, focal tissue bleeding, reactive astrocytosis, and axonal damage. Animals were monitored for intracranial pressure (ICP) and for amino acids in the extracellular fluids. Immunohistochemistry was used to study expression of the gene c-Jun and apoptosis with the terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) technique. Extracellular glutamate, glycine, and taurine increased significantly in the hippocampus within a few hours and remained high after 24 h. Neuronal nuclei in the granule layers of the hippocampus and cerebellum were positive for c-Jun after 24 h. Little immunoreactivity was detected in the cerebral cortex. c-Jun-positive neuronal perikarya and processes were found in granule and pyramidal CA4 layers of the hippocampus and among the Purkinje cells of the cerebellum. Also some microglial cells stained positively for c-Jun. TUNEL reactivity was most intense at 10 days after trauma and was extensive in neurons of the cerebral cortex, hippocampus, and cerebellum. The initial response of the brain after rotational head injury involves brain edema after 24 h and an excitotoxic neuronal microenvironment in the first hour, which leads to extensive delayed neuronal cell death by apoptosis necrosis in the cerebral cortex, hippocampus and cerebellum.

Changes in expression of the proto-oncogene Bcl-2 are well known in the developing brain, with a high expression level in young post-mitotic neurons that are beginning the outgrowth of processes. The physiological significance of the Bcl-2 up-regulation in these neurons is not fully understood. We used a differentiation model for human CNS neurons to study the expression and function of Bcl-2. NT2/D1 human neuronal precursor cells differentiated into a neuronal phenotype in the presence of 10 microM retinoic acid for 3-5 weeks. This concentration of retinoic acid was not toxic to undifferentiated NT2/D1 cells but was sufficient to up-regulate the BCL-2 protein in 6 days. The BCL-2 levels increased further after 3 weeks, i.e. when the cells started to show neuronal morphology. Inhibition of the accumulation of endogenous BCL-2 with vectors expressing the antisense mRNA of Bcl-2 caused extensive apoptosis after 3 weeks of the retinoic acid treatment. The loss of neuron-like cells from differentiating cultures indicated that the dead cells were those committed to neuronal differentiation. Death was related to the presence of retinoic acid since withdrawal of retinoic acid after 16 days of treatment dramatically increased cell surviving. The ability of BCL-2 to prevent retinoic acid-induced cell death was also confirmed in undifferentiated NT2/D1 cells that were transfected with a vector containing Bcl-2 cDNA in sense orientation and exposed to toxic doses (40-80 microM) of retinoic acid. Furthermore, down-regulation of BCL-2 levels by an antisense oligonucleotide in neuronally differentiated NT2/D1 cells increased their susceptibility to retinoic acid-induced apoptosis. These results indicate that one function of the up-regulation of endogenous BCL-2 during neuronal differentiation is to regulate the sensitivity of young post-mitotic neurons to retinoic acid-mediated apoptosis.

Cell specific markers were quantified in the hippocampus, the amygdala/pyriform cortex, the frontal cerebral cortex and the striatum of the rat brain after systemic administration of kainic acid. Neuron specific enolase (NSE) reflects loss of neurons, glial fibrillary acidic protein (GFAP) reflects reactive gliosis, and brain levels of serum proteins measures blood-brain-barrier permeability. While the concentration of NSE remained unaffected in the frontal cerebral cortex and the striatum, their GFAP content increased during the first three days. In the hippocampus and amygdala, NSE levels decreased significantly. GFAP levels in the hippocampus were unaffected after one day and decreased in the amygdala/pyriform cortex. After that, GFAP increased strikingly until day 9 or, in the case of amygdala/pyriform cortex, even longer. This biphasic time course for GFAP was accompanied by a decrease of S-100 during days 1-9 followed by a significant increase at day 27 above the initial level. The regional differences in GFAP and S-100 could result from the degree of neuronal degeneration, the astrocytic receptor set-up and/or effects on the blood-brain barrier.