Oxidative damage and metabolic dysfunction in experimental Huntington's disease: selective vulnerability of the striatum and hippocampus.

The etiology of neuronal death in neurodegenerative diseases, including Huntington's disease (HD), is still unknown. There could be a complex interplay between altered energy metabolism, excitotoxicity and oxidative stress. Unilateral administration of quinilonic acid (QA), NMDA agonist, in rat striatum in a single dose of 150 nM was used as a model of HD. The other two groups of animals were pretreated immediately before QA application with nerve growth factor (NGF) and fibroblast growth factor (FGF), respectively. Control group was treated with 0.9% NaCl in the same manner. Content of total glutathione was not altered in the striatum and hippocampus of QA-treated animals, as well as in the groups pretreated with neurotrophic factors (NF), compared to controls. Content of reduced glutathione, a key antioxidant, was mutually depleted in the striatum and hippocampus of each experimental group. The reduced/total glutathione ratio was decreased in the QA-treated animals, but nearby or over the controls in each structure of the NF-treated groups. These results support the hypothesis that oxygen-free radicals contribute to the excitotoxic neuronal injury, and also that NF could be the potential neuroprotective agents in HD. Moreover, activity of cytochrome c oxidase, the last component in the mitochondrial respiratory chain, was mutually increased in each structure of QA-treated animals. This increase was less pronounced in the NF-treated groups. Striatal lesions led to the loss of tonic inhibitory inputs to the globus pallidus with consequent increase in the activity of GABAergic efferent pallidal neurons, suggesting that NF could functionally repair the altered striopallidal pathway.

[Septic encephalopathy--prognostic value of the intensity of consciousness disorder to the outcome of sepsis]. / Septicka encefalopatija--prognosticka vrijednost intenziteta poremaca svijesti za ishod sepse.

Septic encephalopathy (SE) is a common term indicating the development of signs of progressing cerebral dysfunction and is associated with the presence of microorganisms and their toxins in the blood. Aim of this investigation was to analyze the frequency of this complication considering the consciousness disorders in quantitative sense and prognosis of the survival in patients with SE. The investigation comprised patients (n = 54) with positive hemoculture and signs of septic syndrome by the accepted criteria (fever, clinical signs of infection, respiratory frequency, heart rate, plasma lactate, oliguria). Patients with confirmed cerebral injury, hemorrhage or cerebral ischemia were excluded from the study. Lumbar punction and CT-scan of the brain were performed in all patients in order to exclude visible lesions of cerebral parenchyma and eventual presence of cerebral nervous system (CNS) infection as the causes of sepsis. Results of the investigation demonstrated that in 30 (55%) of patients existed mild consciousness disorder at the level of somnolescence, in 18 (33%) consciousness disorder at the level of sopor and in 6 (11%) consciousness disorder at the level of deep coma. Level of consciousness disorder was in positive correlation with the outcome of sepsis syndrome, which was additionally confirmed by the fact that only in the group of patients with deep coma lethal outcome was observed in 3 cases (50% of this subgroup) regardless of intensive antibiotic, metabolically active and symptomatic therapy. It can be concluded that SE syndrome has a favorable prognosis if macroscopic lesion and dissemination of microorganisms in CNS are not present, and simultaneously it represents changes in metabolic-electrolytic state with early presentation of consciousness disorders that represent clinically significant indicator for sepsis syndrome outcome.

[The closed head injury syndrome--frequency and character of brain parenchyma lesions]. / Sindrom zatvorene povrede glave--ucestalost i karakter lezija mozdanog parenhima.

The aim of this investigation was to determine the frequency and character of the brain parenchyma lesions using CT scan and MR in patients with closed head injury, and afterwards to establish the connection of those changes with clinical parameters (neurological deficit and the degree of consciousness disorder), evaluating the sensitivity and specificity of neuroradiological procedures. The investigation comprised 40 patients with closed head injury. Grade of neurological deficit (Canadian Neurological Scale) and the degree of consciousness disorder (Glasgow Coma Scale) were determined in all patients by the same neurologist. The results revealed significantly higher specificity and sensitivity of brain examination using MR in patients with closed head injury, particularly in the detection of small ischemic and contusion lesions. Likewise, the correlation of positive findings using MR was more significant concerning the grade of neurological deficit and the degree of consciousness disorder compared to the brain examination using CT scan.

Cytochrome C oxidase activity and total glutathione content in experimental model of intracerebral aluminum overload.

Treatment of Wistar rats with aluminum chloride causes astroglial and neuronal cell damage in the selective brain regions of association cortex and hippocampus, seen in patients with Alzheimer's disease. Adult Wistar rats were treated with unilateral intrahippocampal injection of AlCl3 in one single dose of 3.7 g/kg b.w. Control group of animals was treated with 0.9% saline solution likewise. Animals were sacrificed by decapitation seven days after the treatment. Activity of cytochrome C oxidase (COX) and total glutathione content were measured in the ipsi- and contralateral hippocampus and forebrain cortex. Activity of COX was mutually decreased in the hippocampus (ipsi- 30%, contra- 34%), as well as in the forebrain cortex (ipsi- 44%, contra- 47%), compared to controls. These decrease could indicate a deficiency in reducing equivalents with concomitant altered proton gradient and function of electron transport chain, as well as decreased ATP synthesis. Content of glutathione, a clue antioxidative factor, was decreased for about 50% in all examined structures, primary suggesting an impaired regeneration of reduced glutathione. Such distribution of diminished antioxidative defense could be the consequence of the specific brain distribution of transferrin receptors, which was also a main protein carrier for Al. Furthermore, at the cellular level Al could impede glycolysis with consequent decreased production of reducing equivalents which were necessary for glutathione synthesis/reduction, as well as for proton gradient and functionality of electron-transport chain.

Effects of nerve growth factor on antioxidative system in the thalamus of MPTP treated Wistar rats.

1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism is one of the most useful models for the study of that disease. It has been suggested that MPTP-induced neurotoxicity may involve the production of reactive oxygen species. MPTP was applied intracerebrally, unilaterally, in the striatum in single dose of 0.09 g/kg b.w. The second group was treated both with MPTP and nerve growth factor (NGF) in dose of 7 ng/ml. NGF was applied immediately after the neurotoxin. Control group was treated with 0.9% saline solution in the same manner. Animals were decapitated 7 days after the treatment. In the group treated with MPTP, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) was decreased in ipsilateral thalamus, compared to control values as well as to the contralateral thalamus. In the same structures superoxide anion production was increased, compared to controls. Following the application of both MPTP and NGF, the activity of SOD and GSH-Px remained on control values, while the superoxide anion content was decreased, compared to controls. These results indicate a temporal and spatial propagation of oxidative stress and spread protective effects of NGF on the thalamus, the structure that is distant, but very tightly connected with striatum, the place of direct neurotoxic damage.

Significance of de-novo IgD and IgM synthesis in CSF of patients with relapsing-remitting form of disseminated demyelinating disease of CNS.

Disseminated demyelinating disease of the central nervous system (DDD CNS) is immunologically mediated, with confirmed significant intrathecal immunoglobulin production. According to recently known immunopathogenic occurrences and activation of humoral immune response, we have assumed that the presence of oligoclonal immunoglobulins of M and D classes can be confirmed in cerebrospinal fluid (CSF) of patients with DDD CNS. With the aim of its further determination in CSF of relapsing-remitting DDD CNS patients in either remission and relapse phase, respectively, we have confirmed the presence of oligoclonal IgD and IgM bands, the association of this production and the presence of new demyelinating zones found by MRI of endocranium, as the time elapsed from the last relapse until the obtaining of CSF for further analyses. Method of isoelectric focusing with Western blott procedure was used for the confirmation of oligoclonal IgM and IgD bands presence in CSF. Significant presence of intrathecally synthetized oligoclonal IgM and IgD in patients with DDD CNS in exacerbation phase was presented. Almost in all patients in this phase was found at least one indicator of acute phase (positive MRI finding, presence of oligoclonal IgM or IgD bands). Significant decrease of positive findings of oligoclonal Ig bands in CSF was correlated with the time elapsed from the onset of relapse until the obtaining of CSF for the analysis due to short half-life of those Ig in CSF.

The significance of determination of the fraction of creatinine-phosphokinase in patients with acute ischemic brain disease.

Heart and brain vascular diseases are the leading causes of mortality in the world. Cardiac complications can frequently occur during the development of cerebral ischemia. The aim of this study was to establish the possible changes in fractions of creatinine-phosphokinase as the sensitive laboratory index of parenchymal lesion of brain parenchyma and the presence/absence of risk factors for ischemic brain and heart disease. The study comprised 80 patients with acute ischemic brain disease (AIBD), without the history of previous coronary disease. Blood samples were taken in all patients within the first 48 hours from AIBD onset aiming to determine a total (muscular MM) and heart fraction of creatinine-phosphokinase (MB), and brain parenchyma ischemia was confirmed by CT or MR scan of the head. A detailed history of the risk factors for ischemic brain disease (IBD) and ischemic heart disease was taken from all patients with AIBD, and the profile of glycemia and lipid status were determined, and blood pressure was measured 6 times a day. Independent variables in statistical analysis were: age, degree of severity and the side of neurologic event, size of ischemic lesion and maximal values of systolic and dyastolic pressure. Dependent variables were the values of fractions of creatinine-phosphokinase (CPK). Control group (n = 40) comprised patients with neurologic diseases of non-vascular origin. All parameters as well as their interrelations were statistically analyzed. The results revealed significant correlation of the increased levels of CPK of MM and MB fraction with the size and place of ischemic lesion in the right cerebral hemisphere, which was highly significant for MB fraction in the total group of patients with AIBD, and for MM fraction, only for cases of more severe IBD. Highly significant increased values of those fractions were also observed compared to the control group of patients.

Brainstem auditory evoked potentials following head injury.

Brainstem auditory evoked potentials (BAEP) were examined in 40 patients with subjective disorders following closed head injury (CHI), with the established degree of recovery and performed CT-scan of the head. For all BAEP parameters the interval of normality was defined as 3 SD above and below mean value in the control group comprised of 20 healthy subjects. The upper limits of thus defined intervals of normality enabled the formation of four types of findings: type 1--normal finding that was registered in 23 (57.5%) patients; type 2 was a sum of individual findings with the prolonged interpeak latencies, but without the change of relative amplitude V:I--7 (17.5%) recordings; type 3--the findings where the fall of relative amplitude V:I was registered together with the prolongation of interpeak latency. It comprised of 4 (10%) recordings and the type 4 included 6 (15%) individual recordings with registered low RA V:I (0.8 or lower). The explanation of the most probable genesis of registered changes was presented.

Comparative study of the effects of peptidoglycan monomer and structurally related adamantyltripeptides on humoral immune response to ovalbumin in the mouse.

Peptidoglycan monomer, GlcNAc-MurNAc-L-Ala-D-isoGln-mesoDAP(omega NH2)-D-Ala-D-Ala (PGM) originating from Brevibacterium divaricatum and synthetic adamantyltripeptides, diastereoisomers of D,L-(adamant-2-yl)-Gly-L-Ala-D-isoGln (AdTP1 and AdTP2) exhibit immunomodulating activity. An experimental model in the mouse has been established with suboptimal amounts of ovalbumin (OVA) as the immunogen, and parallel testing of adjuvant activity of these three immunomodulators was carried out in Balb/c, C57B16 or CBA mice. Tested compounds (100 or 200 micrograms/mouse) mixed with OVA in saline (50 micrograms/mouse) were administered s.c. Anti-OVA was assayed by ELISA in the sera of mice taken 7 days after the boosters (given on days 14 and 28). The treatment with PGM and one of the diastereoisomers, AdTP2, resulted in significantly higher increase in anti-OVA IgG levels (stimulation index up to 46) with respect to controls and groups treated with AdTP1. The effect of AdTP2 treatment was comparable to that of PGM in most experiments after the first booster, but after the second booster PGM exhibited markedly better effect. PGM and AdTP2 also induced markedly higher levels of IgG1 and IgG2 anti-OVA subclasses than detected in controls and AdTP1 treated mice, indicating that these two immunomodulators might upregulate both Th1-like and Th2-like immune responses.

Plasma exchange in myasthenia gravis and multiple sclerosis.

During the last twenty-year period therapeutic plasma exchange (TPE) was used in the treatment of 68 patients with myasthenia gravis and 61 patients with multiple sclerosis. The therapeutic effects were evaluated on the basis of neurologic deficit changes, electrophysiological findings, necessary laboratory analyses and patient's general conditions. It was shown that the therapeutic effects mosty depended on the nature and stage of the basic disease, adequate selection of the patients and timely applied therapeutic procedure. Significant positive effects of the TPE treatment applied with the anti-inflammatory and immunosuppressive therapy were observed in patients with myasthenia gravis and multiple sclerosis upon clinical findings and some paraclinical tests.

Index of lipid peroxidation and glucose utilization in the cerebrospinal fluid in patients with cerebral infarction.

Cerebral ischemia could be observed as acute metabolic crisis, when oxygen and glucose supply is compromised and synthesis of energy is insufficient. Apart from the excitotoxicity, increased production of reactive oxygen species with consequent lipid peroxidation is also included in neuronal cell damage. Furthermore, these toxic compounds could also be produced during the process of secondary inflammation of ischemic tissue. In the early stage of ischemia, as a systemic response to acute stress, there is an increase in glucose level in cerebrospinal fluid (CSF) and peripheral blood. According to the metabolic crisis and acidosis in ischemic brain tissue we investigated index of lipid peroxidation (ILP) and glucose utilization (IGU) in CSF of 53 patients of both sexes, aged 55-70 years with cerebral infarction. Control group comprised 15 patients with sudden onset of motor deficit subjected to diagnostic lumbar radiculography and suspected on discal genesis. ILP in CSF, as the indicator and sequela of neuronal cell membranes damage, was two fold increased in the acute period of cerebral infarction and maximal values (3.5 times) were noticed 24 hours after the ischemic episode compared to controls. Besides the increase in glucose concentration in peripheral blood and CSF of patients with cerebral infarction, IGU was decreased (37%) with minimal values (32%) 24 hours after the ischemia. These changes indicate that glucose is available but cells are incapable to metabolize it. We concluded that ILP and IGU in CSF of patients with cerebral infarction could be indicators of metabolic dysfunction and neuronal cell damage. Also, these results suggest the significance of polyvalent therapy including antioxidative and antiinflammatory agents in acute phase of cerebral ischemia.

[Neurologic manifestations after surgical interventions]. / Neuroloske manifestacije nakon hirurskih intervencija.

Administration of general anesthesia is rarely accompanied with newly developed postoperative neurological complications. We analyzed postoperative complications after general anesthesia where an urgent neurologic assistance was necessary. The investigation included 120 patients. The same neurologist performed neurologic examination and electroencephalography, and computerized tomography (CT) was performed if necessary. In 96 (80%) patients focal stimulative or destructive phenomena such as epi-seizures or neurologic deficit were not detected by neurologic examination. In 9 (7.5%) patients were detected consciousness crisis. In 6 patients (5%) were registered right extremities weakness with motor dysphasia, which was withdrawn in first 24 hours. In these cases EEG revealed weak activity in theta frequency, above frontoparietal regions, bilaterally. In 6 (5%) patients was registered neurologic deficit of hemiparesis or semi-severe degree with development of ischemic lesion confirmed by CT. In 6 (5%) patients, CT scan revealed the presence of mild brain edema. Also, positive correlation between duration of anesthesia, age and metabolic disorders, specially diabetes mellitus, was found. We concluded that age, type of surgical intervention and duration of general anesthesia had the greatest influence on the development of neurologic disorders during and after general anesthesia, and the presence of metabolic disorders and previous brain damage increase the risk for the onset of these complications.

[Therapeutic value of antioxidants and calcium channel blockers in patients in the acute phase of closed head injuries]. / Terapijska vrijednost antioksidanasa i blokatora kalcijumski kanala kod bolesnika sa zatvorenom povredom glave u akutnoj fazi.

Knowing that uncontrolled calcium signalization with excessive production of reactive oxidative matters is present in case of neurotrauma, aim of the investigation was to establish therapeutic value of combined administration of antioxidants (AO) and calcium channel blockers (CCB) in patients with closed head injury (CHI). Investigation comprised 120 patients with CHI who received AO (vitamins C and E) parenterally during 10 days and CCB (nimodipine), and control group was comprised of 60 patients with CHI who did not receive these medicinals in therapeutic program. We have established the influence of the therapy on neurologic and functional deficiency and consciousness disorder, respectively. Results of the investigation confirmed better recovery of all three observed parameters (degree of neurologic deficiency, degree of functional deficiency and consciousness disorder) in a group of patients receiving AO and CCB, which was statistically significant. It can be concluded that the administration of AO and CCB in patients with CHI in the acute phase should be included into therapeutic program of this significant clinical syndrome.

Oxidative stress indicators are elevated in de novo Parkinson's disease patients.

In order to determine the baseline state of oxidative stress indices in drug-free patients with Parkinson's disease (PD), we analyzed cerebrospinal fluid (CSF) and peripheral blood samples of 34 subjects in disease stages I-III, according to the Hoehn & Yahr scale, compared with controls. The results showed a highly significant increase in malondialdehyde content in CSF (p < 0.001) in the patients with PD. We also found significant differences in peripheral blood parameters between the two groups: malondialdehyde content was increased in patients with PD (p < 0.05), as was the activity of antioxidative defense enzymes, glutathione reductase, Cu, and Zn-superoxide-dismutase (p < 0.05). The production of a highly reactive oxygen species--superoxide radical (p < 0.05) was also increased. These findings indicated an important role of oxidative stress in PD evolution and progress. The increased levels of reactive oxidative species (malondialdehyde content and superoxide radical production) in peripheral blood, and excessive activity of protective enzymatic systems (glutathione reductase Cu, and Zn-superoxide-dismutase) could indicate an additional systemic reaction related to a chronic oxidative stress state in the brain.