Anticonvulsant properties of the cerebrospinal fluid during activation of the antiepileptic system of the brain

Cerebrospinal fluid (CSF) was obtained after 30-40 sessions of daily electrical stimulation of the cat cerebellum vermis. The intraventricular injection of CSF (10 microliters) to Wistar rats increased the latent period of initial seizure manifestations, significantly reduced the number of animals with seizures and reduced the severity of seizures induced by korazol injection (40 mg/kg). Analogous seizure changes were observed in rats after intraventricular injection of CSF (10 microliters) from cats subject to 3-10 electroshock seizure fits. Intraventricular injection of CSF (250 microliters) obtained from cats after electroshock to cats with strychnine-induced epileptic foci in the brain cortex led to the suppression of the epileptic activity. The conclusion was made that different ways of antiepileptic system activation cause the accumulation of endogenous antiepileptic substances in CSF.     

Cerebrovascular damage caused by platelet and leukocyte activation and treatment with neurotroponin]

Effects of neurotropin (a non-protein extract obtained from the exudate of rabbit skin tissue treated with vaccinovirus) on platelet and neutrophil aggregation were studied. In vitro neurotropin reduced platelet and neutrophic aggregability only at high concentration. However, in vivo anti-aggregating effect of neurotropin was more marked. Intravascular aggregation of platelets and leukocytes was induced in cats by selective injection of 4 beta-phorbol-12 beta-myristate-13-acetate (PMA) into carotid artery. It was found that activation of platelets and leukocytes by intracarotid PMA-injection led to a brain ischemia. Regional tissue analysis showed ipsilateral derangement of the cerebral energy state. The values of energy charge in the parietal cortex and caudateputamen of the PMA-injected hemisphere was significantly decreased. Simultaneously the increase of lactate level was observed. Pretreatment with neutropin prevented the development of cerebral energy failure. It is suggested that neutropin appears to be effective for treating cerebrovascular disorders.     

Effect of guanidinoethanesulfonic acid on brain monoamines in the mouse

Guanidinoethanesulfonic acid (GES) is known to induce convulsive seizures when administered intracisternally to rabbits and cats. The effects of GES on behavior, electroencephalographic recording and brain monoamine levels were examined in mice. When GES (900 nmol) was intraventricularly injected into mice, focal clonic movements of the face, vibrissae and ears together with twitching of the limbs were observed 0.5–1 min after the injection. Hypersensitivity was observed up to 7 min after the injection, after which the mice behaved normally. GES also induced sporadic spike discharges on electrocorticogram. The levels of 5-hydroxytryptamine (5-HT) of the GES-injected mice were lower than those of the saline-injected mice in the hippocampus, diencephalon, pons-medulla oblongata and cerebellum 5 min after the injection. No changes in the norepinephrine or dopamine levels were found after the GES injection. The level of 5-hydroxyindoleacetic acid increased in the striatum and cerebellum 5 min after the GES injection. These results suggest that GES-induced convulsive activities enhance the serotonergic neuroactivity in order to suppress the convulsions.     

Effects of neurotropin on seizure activity in picrotoxin kindling]

It is shown that both intracerebral and intraperitoneal neurotropin administration resulted in a decrease of seizure susceptibility of preliminary picrotoxin--kindled rats. On the other hand, neurotropin did not change the course of kindling development. Under conditions of acute picrotoxin--induced seizures it was observed that preliminary cycloheximide (protein-synthesis blocker) administration abolished anticonvulsant properties of neurotropin. It is concluded that anticonvulsant effects of neurotropin are realized via modulation of endogenous peptides synthesis and, in particular, cerulein.     

Morphological changes in the endocrine pancreas of mice after treatment with streptozotocin combined with cyclophosphamide and neurotropin.

The effect of neurotropin (NSP) in combination with streptozotocin (STZ) and cyclophosphamide (CY) on blood glucose and pancreatic histopathology on day 7 and day 14 after the initiation of the treatment was studied in C57Bl/6 male mice. STZ (40 mg/kg) and NSP (1 mg/kg) were applied intraperitoneally on five consecutive days and CY (150 mg/kg)--twice on day 1 and day 3. In single B cells dilatation of the endoplasmic reticulum was found. On day 7 in proximity to some endocrine cells in the mice treated with STZ, STZ + CY + NSP and STZ + CY macrophages were observed. On day 14 lymphocytic infiltration of the islets was demonstrated only in the groups of mice injected with STZ, STZ + CY while in the group treated with the combination STZ + CY + NSP no infiltration was seen. All experimental groups showed no biochemical evidence for hyperglycemia probably due to the mild destruction of a small number of B cells. The results indicate that NSP might possess a restorative action on insulitis induced by multiple low dose streptozotocin administration in mice.     

Elimination of zinc-65 from the brain under kainate-induced seizures

On the basis of the previous evidence that 65Zn concentrations in the brain of EL (epilepsy) mice was affected by induction of seizures, 65Zn movement in the brain was quantitatively evaluated in ddY mice treated with kainate. Six days after intravenous injection of 65ZnCl2, mice were intraperitoneally injected with kainate (10 mg/kg x 6 times in 2 weeks). Myoclonic jerks were observed during treatment with kainate. Twenty days after 65Zn injection, 65Zn distribution in the brain was compared between the kainite-treated and control mice. 65Zn distribution in the brain of the kainate-treated mice was overall lower than in the control mice. 65Zn concentration was significantly decreased in the frontal cortex, hippocampal CA1, thalamus and hypothalamus by treatment with kainate. These results demonstrate that kainate-induced seizures are linked to decreased zinc concentrations in the brain.     

Influence of beta-casomorphin derivatives on chemically and electrically induced seizures

Derivatives of beta-casomorphin(1-5) at a dose level of 1 mumol/kg body weight were tested for their influence on pentylenetetrazol, picrotoxin, or electrically induced seizures after subcutaneous injection in mice. Tyr-Pro-Phe-D-Pro-Gly was found to facilitate pentylenetetrazol-evoked seizures, whereas desTyr derivatives Pro-Phe-D-Pro-Gly and Pro-Phe-Pyr exhibited anticonvulsant properties against those convulsions. The tripeptide was effective only 10 min after application. The beta-casomorphin derivative Pro-D-Phe-Pro-Gly was effective against electrically induced seizures. The protective action of this tetrapeptide lasted for about 5 h. Additionally, we tested the influence of orally administered Pro-Phe-D-Pro-Gly on pentylenetetrazol-induced seizures and Pro-D-Phe-Pro-Gly on electrically induced seizures. Both peptides were effective at a dose of 5 mumol/kg body weight.     

Protective effects of CB1 receptor agonist WIN 55.212-2 in seizure activity in the model of temporal lobe epilepsy

The influence of a low dose (1 microM, 2 microl) of nonselective agonist of cannabinoid CB1 receptor WIN 55.212-2 on seizure activity in different brain structures was investigated in waking guinea pigs. Changes in spontaneous local field potentials and seizure afterdischarges evoked by the electrical stimulation of the perforant path were recorded simultaneously in the hippocampus, entorhinal cortex, medial septal region, and amygdala after a preliminary intraventricular injection of WIN 55.212-2. It was found that WIN 55.212-2 blocked the stimulation-elicited seizures in 80% of tests. A repeated injection of the agonist within 30 days caused an increase in the amplitude of local field potentials and the power of the theta rhythm in all the structures under study. The infusion of kainic acid provoked the onset of status epilepticus in control animals, whereas guinea pigs injected with the agonist (daily, during 25-30 days) did not develop the status. Possible mechanisms of the protective influence of WIN 55.212-2 are discussed.     

Ontogeny of susceptibility to the convulsant, Ro 5-4864, and its relationship to audiogenic seizure susceptibility in inbred mice

The postnatal development of susceptibility to the convulsant effects of Ro5-4864 (4'-chlorodiazepam) was characterized in two inbred mouse strains (DBA/2J and BALB/c ByJ) which as adults differ markedly in their response to this convulsant. Onset of susceptibility to a dose of Ro5-4864 which caused a high frequency of clonic seizures in adults was observed at 10 days of age in DBA/2 mice, but not until 35 days in BALB/c By mice. At 14 days of age an abrupt increase in susceptibility to Ro5-4864-induced tonic seizures was found in DBA/2 but not BALB/c By mice. Both the peak of tonic seizure susceptibility (21 days) and the time course of its subsequent age-dependent decline closely paralleled the change in audiogenic seizure susceptibility in the DBA/2 strain. PK11195 (40 mg/kg) blocked Ro5-4864 (25 mg/kg)-induced, age-dependent tonic seizures but had no effect on clonic seizure induction in the same mice. These observations establish that both the susceptibility to Ro5-4864 in adult mice and the postnatal time course for development of susceptibility to this convulsant are inherently different in these two strains of mice. The lack of coincidence between the developmental onset of susceptibility to Ro5-4864-induced seizures and the onset of supersensitivity to Ro5-4864-induced tonic seizures during the period of peak audiogenic seizure susceptibility in DBA/2 mice implies that more than one neurochemical mechanism is involved in the ability of Ro5-4864 to induce seizures in this strain. However, the blockade of Ro5-4864-induced tonic seizures by PK11195 suggests that peripheral type benzodiazepine receptors may mediate this effect.     

Acupuncture on experimental epilepsies

The therapeutic effect of acupuncture on epilepsies was evaluated in 4 experimental models. 24 acupuncture points were tried. In electroconvulsive threshold model, square wave electrical stimulus of 0.2 msec and 6 Hz was applied through a pair of cotton electrodes at the cornea of mice for 3 sec. The stimulus intensity to induce stun reaction of the mouse was compared. In 86 control animals, the stimulus threshold was 0.70 +/- 0.22 mA. In the acupuncture treated group (N = 80), the threshold was 0.75 +/- 0.14 mA. In maximal electroshock model, the stimulus parameters were 60 Hz, 0.4 sec and 75 mA. The tonic extensor response of the hindlimbs of the mice was observed. 75.7% of the 115 control mice and 77.5% of the 80 acupuncture treated mice were observed to have tonic extensor response. In the focal cortical penicillin model, penicillin was applied at the subpial space over the exposed cortex of 24 cats. After the appearance of repeated spikes in ECoG, acupuncture was performed. In 175 trials the interictal spikes were decreased in 16 times, increased in 82 times. In 99 trials during seizures, the ictal activity was decreased in 4 times, increased in 79 times. In the intravenous penicillin model, high dose penicillin (1,000,000-1,500,000 U/kg) was given to 20 cats. It induced repetitive spikes and frequently even seizure discharges in EEG. Acupuncture was then tried. In 192 instances, acupuncture reduced the spikes in 13, increased the spikes in 103 times. In 74 trials during seizures, the ictal activity was suppressed in 4 times and aggravated in 66 times.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epileptogenesis following Kainic Acid-Induced Status Epilepticus in Cyclin D2 Knock-Out Mice with Diminished Adult Neurogenesis

The goal of this study was to determine whether a substantial decrease in adult neurogenesis influences epileptogenesis evoked by the intra-amygdala injection of kainic acid (KA). Cyclin D2 knockout (cD2 KO) mice, which lack adult neurogenesis almost entirely, were used as a model. First, we examined whether status epilepticus (SE) evoked by an intra-amygdala injection of KA induces cell proliferation in cD2 KO mice. On the day after SE, we injected BrdU into mice for 5 days and evaluated the number of DCX- and DCX/BrdU-immunopositive cells 3 days later. In cD2 KO control animals, only a small number of DCX+ cells was observed. The number of DCX+ and DCX/BrdU+ cells/mm of subgranular layer in cD2 KO mice increased significantly following SE (p<0.05). However, the number of newly born cells was very low and was significantly lower than in KA-treated wild type (wt) mice. To evaluate the impact of diminished neurogenesis on epileptogenesis and early epilepsy, we performed video-EEG monitoring of wt and cD2 KO mice for 16 days following SE. The number of animals with seizures did not differ between wt (11 out of 15) and cD2 KO (9 out of 12) mice. The median latency to the first spontaneous seizure was 4 days (range 2 – 10 days) in wt mice and 8 days (range 2 – 16 days) in cD2 KO mice and did not differ significantly between groups. Similarly, no differences were observed in median seizure frequency (wt: 1.23, range 0.1 – 3.4; cD2 KO: 0.57, range 0.1 – 2.0 seizures/day) or median seizure duration (wt: 51 s, range 23 – 103; cD2 KO: 51 s, range 23 – 103). Our results indicate that SE-induced epileptogenesis is not disrupted in mice with markedly reduced adult neurogenesis. However, we cannot exclude the contribution of reduced neurogenesis to the chronic epileptic state.     

Does vigabatrin possess an anticonvulsant action against pentylenetetrazol-induced seizures in developing rats?

Anticonvulsant action of vigabatrin (300, 600, 900 and/or 1200 mg/kg i.p.), an inhibitor of GABA-transaminase, was studied in a model of motor sezures elicited by pentylenetetrazol. Five age groups of rats (7, 12, 18, 25 and 90 days old) received a s.c. injection of pentylenetetrazol 4, 6 and/or 24 hours after vigabatrin administration. The incidence of minimal, predominantly clonic seizures was not changed in any age group, but their latencies were prolonged in 18- and 25-day-old rats. Generalized tonic-clonic seizures were influenced in a more complex manner. Incidence of these seizures was decreased in 7-day-old rat pups 24 hours after vigabatrin administration. Higher doses of vigabatrin exhibited a similar effect in adult rats at all intervals studied. Specific suppression or at least restriction of the tonic phase was observed in all groups of immature rats, the effect was more marked 24 hours after vigabatrin than at shorter intervals. The anticonvulsant action of vigabatrin, which could be demonstrated mainly against generalized tonic-clonic seizures, varies markedly during development.     

Effects of extracts of different parts of the brain of kindled animals on seizure activity of recipient rats

The peptide-containing fraction was emitted from the hippocampal and ventral mesencephalic region tissue of rats kindled with subconvulsant doses of corazol. Extracts were prepared by the help of hot acetic acid on the stage of generalized clonic-tonic seizure development. The intraventricular injection of VMR-extracts in relatively high dose increased seizure reactions which were induced in intact recipient rats by intraperitoneal corazol injection. The intraventricular injection of the extract in relatively low dose (100 times less) suppressed corazol-induced seizures in recipients. Data are discussed from the point of view of pathological epileptic system formation and the role played by peptides in supporting it's activity during pharmacological kindling.     

Mid-gestation pregnancy termination by the progesterone antagonist aglepristone in queens

The efficacy of aglepristone, a progesterone receptor antagonist, to induce abortion on days 25 and 26 after first mating was investigated in queens. The cats were divided into two groups: aglepristone (10 mg/kg, subcutaneously) was injected twice, 24 h apart, on days 25 and 26 after first mating, into group I queens (n = 23). Group II queens (n = 6) were not treated and served as controls. Termination of pregnancy and expulsion of the fetuses were successful in 20 (87%) queens in group I. The mean interval between the first administration of aglepristone and the beginning of vaginal discharge was 5+/-1 days (range 4-7 days) and the mean duration of abortion, defined as time span from first occurrence of vaginal discharge to expulsion of all fetuses observed by ultrasonography was 1 day in nine cats, 2 days in five cats and in five cats, less than 1 day. Treatment failed in three queens. In one queen treatment resulted in birth (66 days after mating) of two vital kittens. In another case, three macerated fetuses were found intrauterine without vaginal discharge. In one cat, two fetuses were expulsed and two remained intrauterine and were born 66 days after last mating. All group II queens gave birth to vital kittens after a normal pregnancy length. The mean serum P4 concentrations were similar in treated and control animals. The results indicate that aglepristone treatment at day 25 of pregnancy could induce abortion in 87% of the treated queens. Itching at the site of injection right after injection was the only side effect noticed and only in one queen.     

Pronounced increases in brain levels of calcitonin gene-related peptide after kainic acid induced seizures

Changes in immunoreactivity of calcitonin gene-related peptide (CGRP) were investigated in the brains of rats subsequently to seizures induced by intraperitoneal injection of kainic acid (10 mg/kg, i.p.). Increased levels of the neuropeptide were observed in the frontal cortex (increase of 1300% of control value), striatum (900%), dorsal hippocampus (400%) and amygdala/pyriform cortex (135%) three days after injection of the neurotoxin. Intravenous infusion of mannitol (1.5 g/kg, under thiopental anesthesia) which prevents seizures and post-seizure brain damage suppressed the changes in CGRP-like immunoreactivity. Injection of pentylenetetrazol causing generalized motor seizures resulted in no change of CGRP-immunoreactivity after three days. The pronounced but reversible increases of brain CGRP levels suggest a strong but short-lasting activation of the peptide system. The failure of pentylenetetrazol to produce a similar effect and the protective action of mannitol suggest that sustained seizures and/or post-seizure brain damage may be required to produce the rise in peptide levels.     

Dopamine antibodies: effect on the "open field" behavior, pain sensitivity, monoamine levels in the central nervous system, and functional activity of immunocytes in C57Bl/6 mice

Dopamine antibodies (AB) were singularly injected to C57B1/6 mice intraperitoneally. The locomotor activity in the open field was suppressed for 5 days in the majority of the animals. Hyperalgesia revealed 1.5 h and 1 day after the AB injection changed for analgesia on the 5th day. A sharp reduction of the brain level of dopamine and its metabolite was revealed 1 and 5 days after the AB injection, the serotonin content was increased within 1 day and decreased within 5 days after the injection. No action of the AB on cells of the immune system was observed. The possible mechanisms of the AB neurotropic action are discussed.     

Tachyzoite-induced life cycle of Toxoplasma gondii in cats

The tachyzoite-induced cycle of Toxoplasma gondii was studied in 46 cats. Tachyzoites of the M-7741 or Me-49 strain of T. gondii were administered orally to cats by pouring into the mouth or by stomach tube, or by intraintestinal inoculation. Ten weaned cats that had been inoculated with tachyzoites directly in the intestine were killed 1, 3, 6, 9, 12, 15, 18, or 25 days later, and their tissues were studied histologically and bioassayed in mice. Toxoplasma gondii was demonstrable in the blood of 8 cats and in other tissues of all these 10. Four out of five 1- to 8-day-old cats fed tachyzoites by stomach tube became infected with T. gondii, and 1 became ill because of toxoplasmosis. All 19 weaned cats fed tachyzoites (poured into the mouth) became infected, and 6 died of acute toxoplasmosis 9-15 days after being fed T. gondii. Six out of 12 weaned cats fed tachyzoites by stomach tube became infected but were asymptomatic. Overall, 12 out of 26 cats observed for 19 days or more shed oocysts with a prepatent period (pp) of 19 days or more, with the sole exception of 1 cat that shed oocysts with a pp of 5 days. Enteroepithelial stages of T. gondii were not found in any cat before oocysts were shed. Cats shed up to 360 million oocysts in a day, and oocysts were shed for 4-6 days.     

Influence of a new antiepileptic drug ORG 6370 on metrazol-induced seizures in rats during ontogenesis

The anticonvulsant action of a new antiepileptic drug ORG 6370 (Organon International B.V.) was studied in 217 male albino rats aged 7, 12, 18, 25 and 90 days. Epileptic phenomena induced by a subcutaneous injection of a 100 mg/kg dose of metrazol (isolated myoclonic jerk, minimal metrazol seizures and major, generalized tonic-clonic metrazol seizures) were used as a model. ORG 6370 did not influence myoclonic jerks or minimal metrazol seizures in those age groups where they were regularly elicited. In 12-day-old rats pretreatment with ORG 6370 led to the appearance of minimal metrazol seizures, a phenomenon rarely seen under control conditions. Major seizures were suppressed only in adult rats with a 20 mg/kg dose; on the other hand, ORG 6370 exhibited a selective action against the tonic phase of major seizures at all stages of development. The profile of action of ORG 6370 is almost the same as that of phenytoin.     

Effects of purified human interleukin-1 on sleep and febrile response of cats

From cats prepared for chronic polygraphic recordings sleep patterns were obtained for 8 hours after: 1) intracerebroventricular (icv) injection of artificial cerebrospinal fluid (aCSF), day 1; 2) icv injection of interleukin-1 (I1-1), day 2; 3) injection of aCSF, 24 h after injection of I1-1, day 3; 4) injection of aCSF, 48 after injection of I1-1, day 4. Three doses of I1-1 were tested. The dose of 10 nmol slightly prolonged sleep, whereas a dose of 40 nmol totally inhibited sleep. Twenty nmol of I1-1 elicited sleep and increased body temperature. Total sleep (TS) time was significantly increased due to the significant increase in non REM (NREM) sleep as compared to the control day 1. REM sleep was also increased, but this increase did not reach statistical significance. Wakefulness (W) was significantly reduced. At this time the cats were febrile. On day 3, a further significant increase in TS occurred. NREM was significantly increased when compared with day 1, whereas the increase in REM sleep was significant when compared to both day 1 and day 2. At this time body temperature was normal. The increase in REM sleep on days 2 and 3 resulted entirely from the significant increase in the number of REM periods. On day 4, W showed tendency to increase while sleeping time decreased; such tendency suggests that sleep increase caused by I1-1 slowly returns to the control levels. Our results, together with the earlier evidence on somnogenic and pyrogenic action of I1-1, suggest that these actions may be temporarily dissociated.     

Oocyst-induced Toxoplasma gondii infections in cats

To investigate the oocyst-induced cycle with a 21+ day prepatent period, 32 cats were fed 5 x 10(5) to 2 x 10(7) sporulated oocysts of Toxoplasma gondii and necropsied between 4 hr and 41 days thereafter. The presence of the earliest stages in 7 cats was tested in mice. The tissues of 25 cats were studied histologically; 17 were bioassayed by feeding them to cats to determine, by the length of the prepatent period, whether bradyzoites were present. Based on previous studies, a short (3-10 days) prepatent period indicated that bradyzoites were present in an oral inoculum and a long (greater than 21 days) prepatent period indicated the presence of tachyzoites only. Tissues from 14 cats were also bioassayed in mice for the presence of bradyzoites, using their resistance to pepsin as indicator. Six were studied by both methods. Based on these criteria, tachyzoites predominated in extraintestinal organs during the first 14 days after infection. They were found as early as 4 hr in mesenteric lymph nodes where their number reached 10(4) after 6 and 9 days; they were present after 1 day in all levels of the small intestine and after 6 days in the liver, lung, and blood. Bradyzoites were first detected 10 days after oocyst feeding; they predominated by the third week of infection and were present up to 41 days. Enteroepithelial stages were found histologically only in 2 cats, 24 and 41 days after inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)