The role of cell-free DNA measured by a fluorescent test in the management of isolated traumatic head injuries.

BACKGROUND: Traumatic brain injury (TBI) is a major cause of death and disability. In this study a new method to measure cell free DNA (CFD) for the management of TBI is tested. Our hypothesis was that CFD concentrations correlate to the magnitude of brain damage, and may predict the outcome of injured patients. METHODS: Twenty eight patients with isolated head injury were enrolled. Their demographic and clinical data were recorded. CFD levels were determined in patients' sera samples by a direct fluorescence method developed in our laboratory. RESULTS: Mean admission CFD values were lower in patients with mild TBI compared to severe injury (760 ± 340 ng/ml vs. 1600 ± 2100 ng/ml, p = 0.03), and in patients with complete recovery upon discharge compared to patients with disabilities (680 ± 260 ng/ml vs. 2000 ± 2300 ng/ml, p = 0.003). Patients with high CFD values had a relative risk to require surgery of 1.5 (95% CI 0.83 to 2.9) a relative risk to have impaired outcome on discharge of 2.8 (95% CI 0.75 - 10), and a longer length of stay (12 ± 13 days vs. 3.4 ± 4.8 days, p = 0.02). CFD values did not correlate with CT scan based grading. CONCLUSIONS: CFD levels may be used as a marker to assess the severity of TBI and to predict the prognosis. Its use should be considered as an additional tool along with currently used methods or as a surrogate for them in limited resources environment.

A modified human head model for the study of impact head injury.

A recently published finite element (FE) head model is modified to consider the viscoelasticity of the meninges, the spongy and compact bone in the skull. The cerebrospinal fluid (CSF) is simulated explicitly as a hydrostatic fluid by using a surface-based fluid modelling method, which allows fluid and structure interaction. It is found that the modified model yields smoother pressure responses in a head impact simulation. The baseline model underestimated the peak von Mises stress in the brain by 15% and the peak principal stress in the skull by 33%. The increase in the maximum principal stress in the skull is mainly caused by the updation of the material's viscoelasticity, and the change in the maximum von Mises stress in the brain is mainly caused by the improvement of the CSF simulation. The study shows that the viscoelasticity of the head tissue should be considered, and that the CSF should be modelled as a fluid, when using FE analysis to study head injury due to impact.

Increased intracranial pressure is associated with elevated cerebrospinal fluid ADH levels in closed-head injury.

OBJECTIVES: Head injury frequently results in increased intracranial pressure and brain edema. Investigators have demonstrated that ischemic injury causes an increase in cerebrospinal fluid (CSF) levels of antidiuretic hormone (ADH); increased CSF ADH levels exacerbate cerebral edema, and inhibition of the ADH system with specific ADH antagonists reduces cerebral edema. The current study was designed to test the hypothesis that elevated levels of ADH are present in the CSF of subjects with head injury. METHODS: Ventricular CSF and blood samples were taken from 11 subjects with head injury and 12 subjects with no known head trauma or injury. ADH levels were analyzed using radioimmunoassay. Severity of increased intracranial pressure (ICP) was rated in head-injured subjects using a four-point ordinal scale, based on which treatments were necessary to reduce ICP. RESULTS: Subjects with head injury had higher CSF (3.2 versus 1.2 pg/ml; P<0.02) and plasma (4.1 versus 1.4 pg/ml; P<0.02) levels of ADH than did control subjects. In head-injured subjects, CSF ADH levels positively correlated with severity of ICP. DISCUSSION: The results of this study suggest that ADH plays a role in brain edema associated with closed head injury.

Effect of acute inflammatory brain injury on accumulation of morphine and morphine 3- and 6-glucuronide in the human brain.

OBJECTIVE: In animals, central nervous system inflammation increases drug accumulation in the brain partly due to a loss of central nervous system drug efflux transporter function at the blood-brain barrier. To determine whether a similar loss of active drug efflux occurs in humans after acute inflammatory brain injury. DESIGN: Observational human pharmacokinetic study. SETTING: Medical-surgical-neurosurgical intensive care unit at a university-affiliated, Canadian tertiary care center. PATIENTS: Patients with acute inflammatory brain injury, including subarachnoid hemorrhage (n = 10), intracerebral and/or intraventricular hemorrhage (n = 4), or closed head trauma (n = 2) who received morphine intravenously after being fitted with cerebrospinal fluid ventriculostomy and peripheral arterial catheters. INTERVENTIONS: We correlated the cerebrospinal fluid distribution of morphine, morphine-3-glucuronide, and morphine-6-glucuronide with the cerebrospinal fluid and plasma concentration of the proinflammatory cytokine interleukin-6 and the passive marker of blood-brain barrier permeability, albumin. MEASUREMENTS AND MAIN RESULTS: Acute brain injury produced a robust inflammatory response in the central nervous system as reflected by the elevated concentration of interleukin-6 in cerebrospinal fluid. Penetration of morphine metabolites into the central nervous system increased in proportion to the neuroinflammatory response as demonstrated by the positive correlation between cerebrospinal fluid interleukin-6 exposure and the area under the curve cerebrospinal fluid/plasma ratio for morphine-3-glucuronide (r = .49, p < .001) and morphine-6-glucuronide (r = .51, p < .001). In contrast, distribution of morphine into the brain was not linked with cerebrospinal fluid interleukin-6 exposure (r = .073, p = .54). Albumin concentrations in plasma and cerebrospinal fluid were consistently in the normal range, indicating that the physical integrity of the blood-brain barrier was likely undisturbed. CONCLUSIONS: Our results suggest that central nervous system inflammation following acute brain injury may selectively inhibit the activity of specific drug efflux transporters within the blood-brain barrier. This finding may have significant implications for patients with neuroinflammatory conditions when administered centrally acting drugs normally excluded from the brain by such transporters.

Granulocyte colony-stimulating factor does not affect contusion size, brain edema or cerebrospinal fluid glutamate concentrations in rats following controlled cortical impact.

INTRODUCTION: Granulocyte colony-stimulating factor (G-CSF) is an established treatment in the neutropenic host. Usage in head-injured patients at risk for infection may aggravate brain damage. In contrast, evidence of G-CSF neuroprotective effects has been reported in rodent models of focal cerebral ischemia. We investigated effects of G-CSF in acute focal traumatic brain injury (TBI) in rats. METHODS: Thirty-six male Sprague-Dawley rats were anesthetized with 1.2%) to 2.0% isoflurane and subjected to controlled cortical impact injury (CCII). Thirty minutes following CCII, either vehicle or G-CSF was administered intravenously. Animals were sacrificed 24 hours following CCII. Glutamate concentrations were determined in cisternal cerebrospinal fluid (CSF). Brain edema was assessed gravimetrically. Contusion size was estimated by 2,3,5-triphenyltetrazolium chloride staining and volumetric analysis. RESULTS: Dose-dependent leukocytosis was induced by infusion of G-CSF. Physiological variables were unaffected. Water content of the traumatized hemisphere and CSF glutamate concentrations were unchanged by treatment. Contusion volume was similar in all groups. CONCLUSIONS: A single injection of G-CSF did not influence cortical contusion volume, brain edema, or glutamate concentrations in CSF determined 24 hours following CCII in rats. G-CSF, administered 30 minutes following experimental TBI, failed to exert neuroprotective effects.

Ionized magnesium in the cerebrospinal fluid of patients with head injuries.

BACKGROUND: In head injury patients, a decrease in the serum ionized magnesium (iMg) concentration is considered to be related to the severity of the injury, however, this phenomenon is still not completely understood. The cerebrospinal fluid (CSF) iMg concentration has not been well documented under such conditions and, moreover, its normal value has not yet been established. We hereby intended to investigate the role of the iMg concentration and other parameters in both the serum and CSF of head injury patients and identify any relationship with other parameters. MATERIALS AND METHODS: The subjects consisted of head injury patients without any other serious injuries. Ten healthy volunteers were selected as control subjects. Arterial blood and CSF specimens were simultaneously obtained and measured. We measured the Glasgow Coma Scale scores (GCS), the intracranial pressure (ICP), pH, po2, pco2, sodium, potassium, iCa, iMg, glucose, lactate, urea nitrogen. All data are expressed as the mean+/-SD and the units of iMg and iCa (corrected under pH 7.40) are given in mmol/L. RESULTS: In the healthy subjects, the iMg concentration in the serum/CSF was 0.48 +/- 0.02 / 0.66 +/- 0.14, and iCa was 1.14 +/- 0.05 / 0.94 +/- 0.07. The GCS of the 15 head injury subjects at examination was 8.7 +/- 4.5. When the subjects were divided into 3 groups according to the GCS level (3 and 4, 5-8, and > or =9) at the time of examination, the serum iMg concentration was thus found to be related to the severity of injury based on the GCS level (p = 0.028), but not the CSF iMg concentration (p = 0.89). No relationship was observed between the iMg concentration in the serum and CSF when all specimens were compared, but an extremely close correlation was seen in the group with GCS 3 and 4 (p < 0.0001, r = 0.995), although no such correlation was seen in the other 2 groups (p = 0.12, r = -0.56 in the group with GCS 5-8, and p = 0.26, r = -0.35 in the group with GCS > or = 9). There was a significant correlation between the serum iMg and iCa (p = 0.0093, r = 0.47), and also between the CSF iMg and iCa concentrations (p < 0.0001, r = 0.67). CONCLUSION: The serum iMg concentration has been suggested to possibly affect the neurologic state through CSF iMg in patients with the most severe head injury. In patients with moderate or mild head injuries, however, the ionized magnesium concentration is also probably associated with the degree of neurologic deficit based on the ionized calcium level. The CSF and serum ionized magnesium dissociation may thus result from the slow movement of ionized magnesium through the blood brain barrier.

[Spinal fluid in central nervous system diseases]. / Issledovanie likvora pri zabolevaniiakh tsentral'noi nervnoi sistemy.

Erythrocytes, extracellular hemoglobin and bilirubin level, a quantity of medium mass molecules and catalase activity were determined in the spinal fluid of 32 patient with closed craniocerebral injury of the various degree 24 hours after accident. The extracellular hemoglobin and the bilirubin level were shown to appear in cases with cerebral contusion in association with bleeding only, failed to reflect state severity degree at the early stage after injury. As the number of medium mass molecules increases significantly only in the patients with severe contusions, this index can be used for description of the severity of the process and for evaluation of the deterioration of the clearance of proteolytic products resulting in endogenous intoxication of the central nervous system. The spinal catalytic enzyme activity reflects the severity of the closed cerebrocranial trauma most objectively and can be used as a subarachnoid hemorrhage marker.

Elevated intracranial IL-18 in humans and mice after traumatic brain injury and evidence of neuroprotective effects of IL-18-binding protein after experimental closed head injury.

Proinflammatory cytokines are important mediators of neuroinflammation after traumatic brain injury. The role of interleukin (IL)-18, a new member of the IL-1 family, in brain trauma has not been reported to date. The authors investigated the posttraumatic release of IL-18 in murine brains following experimental closed head injury (CHI) and in CSF of CHI patients. In the mouse model, intracerebral IL-18 was induced within 24 hours by ether anesthesia and sham operation. Significantly elevated levels of IL-18 were detected at 7 days after CHI and in human CSF up to 10 days after trauma. Published data imply that IL-18 may play a pathophysiological role in inflammatory CNS diseases; therefore its inhibition may ameliorate outcome after CHI. To evaluate the functional aspects of IL-18 in the injured brain, mice were injected systemically with IL-18-binding protein (IL-18BP), a specific inhibitor of IL-18, 1 hour after trauma. IL-18BP-treated mice showed a significantly improved neurological recovery by 7 days, accompanied by attenuated intracerebral IL-18 levels. This demonstrates that inhibition of IL-18 is associated with improved recovery. However, brain edema at 24 hours was not influenced by IL-18BP, suggesting that inflammatory mediators other than IL-18 induce the early detrimental effects of intracerebral inflammation.

Nitric oxide (NO) metabolism in the cerebrospinal fluid of patients with severe head injury. Inflammation as a possible cause of elevated no metabolites.

BACKGROUND: This article investigates nitric oxide (NO) metabolism following severe head injury (SHI). We wished to clarify the alterations of NO metabolism end products that is associated with SHI, and to delineate the role of inflammation in this process. METHODS: In a prospective study, we simultaneously measured the concentrations of NO metabolites and interleukin-8 (IL-8) in the ventricular cerebrospinal fluid (CSF) of 11 patients who had suffered SHI. The CSF concentrations of nitrite (NO(-)(2)) and nitrate (NO(-)(3)) combined, and of IL-8 were measured during the following four time periods post-trauma: 6 to 10, 20 to 28, 40 to 56, and 64 to 74 hours. Levels were measured using the corresponding kits. RESULTS: Compared to the ventricular CSF control values, all of our SHI patients had significantly elevated CSF levels of NO(-)(2) plus NO(-)(3) (NO(-)(2) + NO(-)(3)) and IL-8 during all periods tested. CSF NO(-)(2) + NO(-)(3) and IL-8 concentrations reached their maximums simultaneously at 20 to 28 hours following trauma (Spearman's rank correlation = 0.609, p < 0.05), and NO(-)(2) + NO(-)(3) levels were significantly higher than those measured at 6 to 10, 40 to 56, and 64 to 74 hours. [Nitrite-nitrate concentrations: 6-10 hours: 19.22 +/- 6.75, 20-28 hours: 25 +/- 6.2 micromol/l, 40-56 hours: 19.82 +/- 4.47, and 64-74 hours: 19.72 +/- 4.61 micromol/l, (p < 0.05). IL-8 concentrations: 6-10 hours: 3,232 +/- 2,976.2, 20-28 hours: 3,458.45 +/- 3,048 pg/mL, 40-56 hours: 2,616.41 +/- 2,539.21, 64-74 hours: 1,388.88 +/- 1,216.7 pg/mL, (p < 0.001).]. This simultaneous surge in NO(-)(2) + NO(-)(3) and IL-8 in the initial 24 hours post-traumatic indicated that inflammation secondary to SHI increased the rate of NO metabolism, resulting in higher levels of metabolites in the CSF. CONCLUSION: In patients with SHI, CSF concentrations of the dominant metabolites of NO are elevated in the first 3 days after trauma. A similar concurrent spike in the CSF level of IL-8, a marker of acute inflammatory response, can also be demonstrated. These data indicate that the predominant cause of the higher CSF NO(-)(2) + NO(-)(3) concentrations observed in SHI is most likely inflammation.

Increased interleukin-12 levels in human cerebrospinal fluid following severe head trauma.

An overwhelming intracranial inflammatory response occurs as a consequence of severe head trauma, leading to cerebral edema and secondary brain injury. Cytokines are important mediators of post-traumatic cerebral inflammation. In the present study, levels of interleukin-12 (IL-12), a pro-inflammatory cytokine which activates cellular immune response mechanisms, were measured by ELISA in 140 matched serum and ventricular cerebrospinal fluid (CSF) samples from ten patients with severe traumatic brain injury. The mean IL-12 CSF levels were significantly elevated in all patients in the course of 14 days after trauma, compared to CSF samples from 15 control patients. Assessment of the IL-12 CSF/serum ratio and of the blood-brain barrier function, using the CSF/serum albumin ratio, suggest that elevated IL-12 CSF levels might be in part derived from intracerebral cytokine synthesis.

[Repeated closed craniocerebral trauma]. / Povtornaia zakrytaia cherepno-mozgovaia trama.

As many as 446 patients with repeated craniocerebral injury and 386 patients with primary injury, analogous in respect of severity, were subjected to clinical and physiological examinations. On comparison of the results of examining the above two patients' groups it has been revealed that repeated brain injury may be characterized by its own features and runs a graver course as compared to primary injury. This manifests by more pronounced and persistent general cerebral and focal symptoms, disorders of the dynamics of the CSF toward hypotension, vegetative disorders in the form of lability and asymmetry of arterial pressure, thermal asymmetry, prolongation of the time of the thermoregulation vascular reflex and resolution of the blister according to the McClure-Aldrich test, and so forth. According to the therapeutic indications, 204 patients with repeated injury underwent pneumoencephalography. Manifest alterations in CSF-containing spaces, the intensity of which depended on the number of injuries, were revealed in 90.2% of them early after the injury. A special complex of pathogenetic therapy provided to 212 patients with repeated injury allowed attaining more favourable results as compared to 234 analogous patients who received routine treatment.

CSF neurotransmitter metabolites and short-term outcome of patients in coma after head injury.

The main metabolites of the neurotransmitters noradrenaline, dopamine, and serotonin, methoxy-hydroxyphenylglycol (MHPG), homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5HIAA) respectively, were estimated by HPLC with electrochemical detection in CSF samples from 24 patients in coma after head injury, 1 to 12 (mean 3.0) days from accident, and from 24 age- and sex-matched subjects undergoing myelography for possible herniated disk. Analysis of variance with age as covariate, revealed significantly elevated levels of all three metabolites in the patients group. The concentrations of 5HIAA were negatively correlated to the score in the Glasgow Coma Scale. Fourteen patients who recovered with no or minor neurological deficits, had significantly lower CSF 5HIAA levels compared to the ten patients who had a bad outcome (death), while there were no differences regarding HVA or MHPG concentrations. The possibility of a connection of the high neurotransmitter turnover during coma to the development of post-traumatic depression is discussed.

Behavioural and psychosocial sequelae of severe closed head injury and regional cerebral blood flow: a SPECT study.

Thirty six patients (31 male, 5 female) who had suffered severe closed head injury were re-examined at an average of 39.3 (SD 12.8, range 7-66) months after the injury. Behavioural symptoms were measured using the Giessen test. The relatives' reports were used for data analysis to ensure that results were valid. The neurophysical impairment subscale of the Glasgow assessment schedule was completed by two neurologists, and the number connection test was completed by each patient. The adjective mood scale was completed by each relative. All patients were investigated by single photon emission computerised tomography (SPECT). Exploratory factor analysis using the principal components method was carried out separately for SPECT results and psychological measures and correlations were sought between the resulting factors. Factor analysis of the data from the Giessen test identified social isolation, disinhibition, and aggressive behaviour as major components of post-traumatic personality changes; it indicates that these behavioural features are independent of the level of neurological and neuropsychological impairment, which loaded on a single independent factor. Relatives' psychic health seemed to be relatively resistant to physical and cognitive disability and was mainly affected by disinhibitive behaviour. The highest correlation was between frontal flow indices and disinhibitive behaviour (p less than 0.01): the severity of disinhibition increased with lower frontal flow rates. There was a significant but somewhat weaker correlation (p less than 0.05) between flow indices of the left cerebral hemisphere and social isolation. Low flow values of the right brain regions were related to aggressive behaviour (p less than 0.05). Neurological and cognitive impairment correlated negatively with the thalamus; worse neurological and cognitive performance indicate by raised scores on the neurophysical scale and on the number connection test was associated with low thalamic flow values. The results support the importance of lesion location in the production of post traumatic behavioural disorders.