Could phthalates exposure contribute to the development of metabolic syndrome and liver disease in humans?

In the study, 305 patients of both genders were enrolled and divided into three groups: obese (BMI > 30 kg/m2), patients who were diagnosed type 2 diabetes mellitus (T2DM), and control, normal weight healthy volunteers. At least one of ten different phthalate metabolites was determined in the urine samples of 49.84% all enrolled participants. In the obese subgroup, the sum of all urinary phthalate metabolites was positively associated with TG levels (p = 0.031) together with derived TC/HDL and TG/HDL ratios (p = 0.023 and 0.015), respectively. Urinary MEP concentration was positively correlated with the HOMA-IR in T2DM subgroup (p = 0.016) while in the control subgroup, log10MEP levels were negatively correlated with total cholesterol (p = 0.0051), and LDL serum levels (p = 0.0015), respectively. Also, in the control subgroup, positive linear correlations between urinary log10MEP levels and TyG and TYG-BMI values (p = 0.028 and p = 0.027), respectively, were determined. Urinary MEHP levels were associated with glucose serum levels (p = 0.02) in T2DM subgroup, while in the control HDL values were negatively associated with log10MEHP (p = 0.0035). Healthy volunteers exposed to phthalates had elevated AST levels in comparison to non-exposed ones (p = 0.023). In control subgroup, ALT and AST values were increased (p = 0.02 and p = 0.01, respectively) in MEP exposed while GGT levels were enhanced (p = 0.017) in MEHP exposed in comparison with non-exposed. Combined phthalates influence on glucose and lipid metabolism may increase the possibility for NAFLD and insulin resistance development among exposed individuals.

Intestinal intussusception due to a pyogenic granuloma.

Pyogenic granuloma (PG), also known as lobular capillary hemangioma, is a benign vascular tumor, most commonly arising on the skin and the oral mucosa. Gastrointestinal localization of PG, except for the oral cavity, is exceptionally rare. We describe a case of ileal PG occurring in a 13-year-old girl, presenting with intestinal obstruction. Histological examination revealed proliferation of capillary-sized vessels, with prominent intravascular component, involving the entire thickness of the intestinal wall. Immunohistochemistry showed positivity for CD31, CD34 and von Willebrand factor, whereas immunostaining for glucose transporter-1 protein (GLUT1) and for human herpes virus 8 (HHV-8) was negative. We suggest that PG should be considered in the differential diagnosis of childhood gastrointestinal polypoid lesions.

Ontogenetic study of metaphit-induced audiogenic seizures in rats.

Ontogenetic differences in susceptibility to metaphit (1-(1-(3-isothiocyanatophenyl)cyclohexyl)-piperidine)-induced audiogenic seizures were examined in young, developing (ages: 12, 18, and 25 days) and adult (90 days old) Wistar albino rats. Metaphit was injected in a dose of 10 mg/kg i.p. and animals were subjected to intense audio stimulation (100 +/- 3 dB, 60 s) at hourly intervals after administration. Audiogenic seizures (AGS) were scored according to a four point descriptive rating scale (0-3). AGS were elicited in all age groups; they were induced for 12, 15, 15, and 30 h in 12-, 18-, 25-day-old, and adult rats, respectively. Younger animals reached a peak incidence and severity of seizures before adult rats. Twenty-five-day-old rats showed greatest incidence and severity of seizures, and shortest latency. Twelve-day-old animals had longest latencies. Besides audiogenic seizures, we observed convulsions induced by metaphit only in the form of running episodes, forelimb clonus, clonic convulsions, and rearing. Results suggest that young rats develop metaphit-induced sound seizures more rapidly, but that adults have longer period of seizure susceptibility. Different susceptibility to seizures is probably due to changes in excitatory and inhibitory pathways, while maturation of blood-brain barrier is less probable, since metaphit has a lipophilic nature.

Antiepileptic activity of delta sleep-inducing peptide and its analogue in metaphit-provoked seizures in rats.

PROBLEM: Previous studies have shown that humoral, endogenous and somnogenic, delta sleep-inducing peptide (DSIP) has influence on insomnia, pain, adaptation to stress, epilepsy, etc. We investigated the potential of DSIP and its analogue DSIP-12 (a nonapeptide with alanine in position 2 of DSIP molecule substituted by beta-alanine) to antagonize metaphit (1-[1(3-isothiocyanatophenyl)-cyclohexyl]piperidine) induced generalized, reflex audiogenic seizures in adult male Wistar albino rats. METHODS: The rats divided in four groups received (i.p.): saline; metaphit; metaphit+DSIP; and metaphit+DSIP-12, respectively. Metaphit-treated animals displaying seizure in eight previous tests received DSIP or DSIP-12 and afterwards audiogenic stimuli were applied at hourly intervals for the next 30 h. The animals were exposed to sound stimulation 60 min after metaphit administration and further on at hourly intervals. Incidence and severity of seizures were behaviorally analyzed. Selected EEGs and power spectra were recorded and analyzed. RESULTS AND CONCLUSIONS: Metaphit led to hypersynchronous epileptiform activity (polyspikes and spike-wave complexes) and increased power spectra 0.5-30 h after the treatment. Severity of metaphit seizures increased with time to reach the peak 7-12 h after injection. DSIP and DSIP-12 significantly (*P<0.05 and **P<0.01) increased in delta and theta frequency bands and decreased the incidence, mean seizure grade and duration of metaphit convulsions. The results suggest that DSIP and DSIP-12 may be considered as potential antiepileptics in the animal model, DSIP-12 being more efficient than DSIP.

Delta sleep-inducing peptide and its tetrapeptide analogue alleviate severity of metaphit seizures.

The effects of delta sleep-inducing peptide (DSIP) and its tetrapeptide analogue, DSIP(1-4), on metaphit-induced audiogenic seizures were studied. Five groups of adult male Wistar rats were intraperitoneally treated with (1) saline, (2) metaphit, (3) DSIP, (4) metaphit+DSIP and (5) metaphit+DSIP(1-4). To examine blocking effects of DSIP and its analogue on fully developed metaphit seizures, the last two groups were injected after the eight audiogenic testing. The rats were stimulated using electric bell (on the top of the cage, generating 100+/-3 dB and frequency 5-8 kHz, for 60 s) 1 h after metaphit and afterwards at hourly intervals during the experiment. For EEG recordings and power spectra, three gold-plated screws were implanted into the skull. In metaphit-treated animals, EEGs appeared as polyspikes and spike-wave complexes while the power spectra were increasing for 30-h period. The incidence and severity of metaphit-induced audiogenic seizures reached peak value 7-12 h after the injection. Both DSIP and DSIP(1-4) significantly increased power spectra of delta waves and decreased incidence of seizures, mean seizure grade and tonic component of metaphit-induced convulsions. Taken together, these results suggest that DSIP and its analogue DSIP(1-4) should be considered as potential antiepileptics.

Effects of manipulation of N-methyl-D-aspartate receptors on imipenem/cilastatin-induced seizures in rats.

BACKGROUND & OBJECTIVES: Epileptic seizures have been reported in patients on imipenem/cilastatin (Imi/Cil) therapy. To investigate contribution of N-methyl-D-aspartate (NMDA) receptors in inducing imipenem/cilastatin (Imi/Cil) seizures, the effects of competitive NMDA antagonist, APV [(+/-)-2-amino-5-phosphonovaleric acid], non-competitive NMDA antagonist remacemide [(+/-)-2-amino-N-(1-methyl-1,2-diphenylethyl)-acetamidel, and glycine receptor partial agonist HA-966 [(+/-)-(3-amino-1-hydroxypyrrolid-2-one)] on electroencephalographic (EEG) activity and behaviour were studied in rats. METHODS: Adult male Wistar albino rats were implanted with electrodes and cannulae were placed into the right lateral ventricle. Animals were divided into five groups: (i) saline (icv)+Imi/Cil (ii) APV (0.2 micromol)+Imi/Cil, (iii) APV (0.4 micromol)+Imi/Cil, (iv) remacemide (100 mg/kg, ip)+Imi/Cil, and (v) HA-966 (200 microg, icv)+Imi/Cil. The drugs were administered 30 min before icv injection of Imi/Cil (100/100 microg), and their effects on incidence of seizures, latencies to EEG changes and convulsions, severity, lethality and time to lethal outcome were studied. RESULTS: Imi/Cil provoked complete seizure response in all rats and all animals died within 10-18 min after the injection. EEG epileptiform activity preceded behavioral seizures. Clonic-tonic seizures were characterized by continuous bursts of high frequency high amplitude spikes in the EEG. The dose of 0.2 micromol of APV prolonged only the latency to the first EEG changes, while 0.4 micromol dose significantly influenced all seizure parameters. HA-966 increased only the latency to Imi/Cil-induced convulsions, while remacemide had no significant effect on seizure parameters. INTERPRETATION & CONCLUSION: The results suggested that excitatory neurotransmission contributed to the generation and/or propagation of Imi/Cil-induced seizures in rats, and that the effects of NMDA antagonists depended on a particular binding site within the NMDA receptor complex, and affinity to that site.

The effects of phenytoin and phenobarbital on seizures induced by imipenem/cilastatin in rats.

The effects of phenytoin (PHT) and phenobarbital (PHB) on EEG activity and behavior was studied in the model of epilepsy induced by intracerebroventricular (i.c.v.) administration of imipenem/cilastatin (Imi/Cil). Under intraperitoneal (i.p.) sodium pentobarbital anesthesia adult male Wistar albino rats were implanted with electrodes and cannulas were placed into the right lateral ventricle. Animals were divided into groups: 1) Imi/Cil (100/100 microg, i.c.v.), 2) PHT (40 mg/kg) + Imi/Cil (100/100 microg, i.c.v), 3) PHT (80 mg/kg) + Imi/Cil (100/100 microg, i.c.v.), 4) PHT (160 mg/kg) + Imi/Cil (100/100 microg, i.c.v.), 5) PHB (50 mg/kg) + Imi/Cil (100/100 microg, i.c.v.), and 6) PHB (80 mg/kg) + Imi/Cil (100/100 microg, i.c.v.). PHT and PHB were injected intraperitoneally (i.p.) 1 h before Imi/Cil. Seizures were scored according to the scale: 0--normal behavior; 1--twitching, 2--head nodding, forelimb clonus, 3--rearing, and 4--clonic-tonic convulsions. Imi/Cil provoked maximal seizures in all animals, and all rats died 10 - 18 min after the injection. Epileptiform activity preceded behavioral seizures. Clonic-tonic seizures were associated with continuous bursts of high-frequency high-amplitude spikes in the EEG. PHT and PHB suppressed Imi/Cil-induced seizures dose-dependently. PHB reduced epileptiform discharges during behavioral seizures elicited by Imi/Cil, while PHT had no effect on EEG epileptic phenomena. These results suggest that PHT acts as anticonvulsant, and PHB as anticonvulsant and antiepileptic agent in the model of epilepsy induced by Imi/Cil.

[The quality of life in patients with diabetes mellitus type 2].

BACKGROUND/AIM: Through its vrious activities, World Health Organization (WHO) contributed to increasing the understanding of the concept of quality of life. People with diabetes have a lower quality of life than people without chronic illnesses. The aim of this study was to examine the differences in the quality of life, related to health, in patients with diabetes mellitus (DM) type 2 by age, gender and type of therapy. METHODS: We performed a cross-sectional study at the outpatient department of the Clinical Center in Novi Sad and the Health Center Ruma-General Practice. The group consisted of 90 patients with DM type 2, 41 men and 49 women. The age of respondents was from 40 to 80 years and they were classifed into four groups according to the ten-year age intervals. We applied WHO Quality of life questinnaire--BREF 100 composed of four domains: physical health, psychological health, social relationships and environment. The general questionnaire asks questions about socio-demographic data, duration of diabetes, the last value of blood glucose and glycosylated hemoglobin, training for self-control and its implementation, informing patients about their disease, therapy and its impact on daily activities and the presence of comorbidity. In statistical analysis the following tests were used: Student's t-test, F-test, ANOVA (one way). RESULTS: The average duration of DM type 2 was 11.2 +/- 9.2 years. Most of the patients (76%) were trained to self-control and 91% received enough information about their disease. Oral hypoglycemic preparations were used by 49%, insulin by 21%, and oral drugs and insulin by 29% patients while 1% were on a special regime of a diet therapy. Daily activities were performed without difficulties by over 29%, with some difficulties by 41% and 30% of patients who could not perform daily activities. The patients with DM type 2 had significantly lower scors in all 4 domains of quality of life (physical health, psychological health, social relations, environment). The biggest influence was on physical domains (51.31). Education level had an imact on physical and psychological domains. Comorbidity was found in 83% of the respondents. The most common were: arterial hypertension (63%), chronic cardiovascular disease (46%), neuropathy (23%), impaired vision 24%, elevated blood lipids (39%) and amputation of toes or feet (2.2%). The average value HbA1c in the group with comorbidity was 8.47% and in the group without comorbidity 6.46%. The subjects with comorbidity had low quality of life assessment in relation to the group without comorbidity: the domain of physical health (45.64 vs. 79.66), psychological health (50.3 vs. 76.86), social relations (52.97 vs. 75.46) and environment (52.7 vs. 75.06). CONCLUSION: Diabetes mellitus type 2 has negative influence on the quality of life. It contributes to the presence of comorbidity. The occurrence of comorbidity was associated with higher glucosylated HbA1c values. There was no difference in the assessment of quality of life regarding gender, age, or the type of therapy used. The quality of life was assessed as low in patients with comorbidity. However, certain personality characteristics play a decisive role in self-evaluation.

[Role of the chosen general practitioner in educating women on the importance of regular gynecological examinations].

INTRODUCTION: Since regular screening is the best way of preventing the development of cervical cancer, the objective has been set to assess the motivation of women to have regular gynecological examinations and to estimate the role of the chosen general practitioner. MATERIAL AND METHODS: The survey was performed on the basis of the prospective study done at the Health Centre "Novi Sad" in 2009 during the systematic regular examinations carried out by general practitioners. RESULTS: It was found that 60.8% of the examined women had regular checkups; 21.5% visited their doctor once in the period of two to five years and 4.9% had undergone the examination in a period > 10 years, whereas 1.9 women had never had an examination. Other examinees had occasional check-ups with various time laps between them. DISCUSSION: The reasons for not visiting a gynecologist were fear of the examination, absence of discomforts and lack of time. However, 87.2% of the examinees visited a gynecologist after they had been advised to do so by their general practitioner. Gynecological finding was good in 87.6% of the women, 3.4% were found to have carcinoma and 8.9% had some other abnormal finding. CONCLUSION: According to the obtained results, it has been concluded that the chosen general practitioner has a very important role in motivating women to have regular gynecological examinations and in educating them on the risk factors for developing malignant diseases and on the possible prevention.

Effect of gluten-free diet on the growth and nutritional status of children with coeliac disease.

INTRODUCTION: Gluten-free diet (GFD) presents the basis of coeliac disease (CD) treatment. If strictly applied, the disorders of the small bowel mucosa and other disease signs rapidly resolve. OBJECTIVE: The goal of the study was to evaluate the effect of GFD on the growth and nutritional status of children with the classical form of CD. In addition, we analyzed the differences between these parameters with the duration and the patients' compliance with GFD. METHODS: The study goals were achieved on a sample of 90 children, 56 female and 34 male, aged 0.5-7.5 (1.53 +/- 1.05) years, with the classic CD diagnosed on the basis of typical pathohistological findings of the small bowel mucosa and clinical recovery of patients on GFD. The duration of the patients' follow-up was 1.08-8.75 (3.03 +/- 1.14) years, i.e. until the age of 2.5-15 (4.59 +/- 1.78) years. The initial and control values of body height (BH) in relation to matched values for age and gender were expressed in percentiles, while the deviation in body weight (BW) for the matched values of height and gender was expressed in percentages. The referent haemoglobin (Hb) rate in blood, as a laboratory indicator of nutritional status in children aged up to 5 years was > or = 110 g/L, and for those aged above 5 years it was > or = 115 g/L Compliance with GFD was based on the pathohistological findings of the small bowel mucosa or determination of tissue transglutaminase. RESULTS: Over the studied period, the effect of GFD was highly significant, both on the increase of BH percentiles (37.62 +/- 26.26 vs. 57.22 +/- 25.29; p < 0.001), and on the decrease of BW deficit 11.58 +/- 10.80 vs. 0.89 +/- 8.194; p < 0.001). After the treatment period, none of the children showed slowed growth rate or BW deficit above 20%, while BW deviation ranging between 10-20% in relation to the referent values was registered in 17 (18.19%) and the excess of over 20% in 2 patients. In 86 (95.56%) patients, control Hb values in blood were normal, while mild anaemia was registered in 4 patients, all compliant with GFD. The difference between the compliant and non-compliant patients with GFD was not detected either in BH percentiles (p = 0.586) or in BW percentage deviation as compared to standard values (p = 0.516) or in blood Hb values (p = 0.445). In addition, differences between the children on GFD lasting over and below 3 years were not detected either in BH percentiles (p = 0.915) or in BW deviation percentages in relation to the ideal rate (p = 0.476). CONCLUSION: GFD applied for 1-3 years has a highly significant effect on the growth rate and nutritional status of children with the classical form of CD. Significant differences in these parameters of the disease were not detected between strictly compliant and non-compliant patients on GFD.

[Delta sleep-inducing peptide and its analogues alleviate the severity of metaphit-induced audiogenic seizures in rats].

INTRODUCTION: We investigated the potential of delta sleep-inducing peptide (DSIP) and its analogue DSIP-12 (a nonapeptide with alanine in position 2 of DISP molecule substituted by beta-alanine) and tetrapeptide analogue DSIP1-4, to antagonize metaphit(1-1 (3-isothiocyanatophenl)-cvyclohexyl piperidine) induced generalized reflex audiogenic seizures in adult male Wistar albino rats. MATERIAL AND METHODS: Five groups of adult male Wistar rats were intraperitoneally treated with: (1) saline; (2) metaphit: (3) metaphit + DSIP, (4) metaphit + DSIPI-4 and (5) metaphit + DSIP-12. To examine the blocking effects of DSIP and its analogues on fully developed metaphit seizures, in the last three groups they were administered 8h after metaphit injection. The rats were stimulated using an electric bell (1003 dB, 5-8 kHz, 60 s) one hour after metaphit injection and afterwards at hourly intervals during the experiment. For EEG recordings and power spectra three gold-plated screws were implanted into the skull. RESULTS: In metaphit-treated animals EEGs appeared as polyspikes and spike-wave complexes, while power spectra were increasing. The incidence and severity of netaphit-induced audiogenic seizures reached a peak value at 7-12 h after injection. Both DSIP and DSIP analogues significantly increased power spectra of delta waves and decreased the incidence of seizures, as well as mean seizure grade and tonic component of metaphit-induced convulsions. CONCLUSION: Taken together, these results suggest that DSIP and its analogues should be considered as potential antiepileptic agents.

Anticonvulsant, but not antiepileptic, action of valproate on audiogenic seizures in metaphit-treated rats.

1. The blocking effects of valproate (2-propylpentanoic acid), a standard anti-epileptic drug, on metaphit (1-[1-(3-isothiocyanatophenyl)-cyclohexyl]-piperidine)-induced audiogenic seizures as a model of generalized, reflex audiogenic epilepsy in adult Wistar male rats were studied. 2. Rats were stimulated using an electric bell (100 +/- 3 dB, 5-8 kHz, 60 s) 60 min after i.p. metaphit (10 mg/kg) injection and afterwards at hourly intervals. For power spectra and electroencephalograph (EEG) recordings, three gold-plated screws were implanted into the skull. Different doses of valproate (50, 75 and 100 mg/kg) were injected i.p. into rats with fully developed metaphit seizures after the eighth audiogenic testing. 3. In metaphit-treated animals, the EEG appeared as polyspikes, spike-wave complexes and sleep-like patterns, whereas the power spectra were increased compared with the corresponding controls. 4. Valproate reduced the incidence and intensity of convulsions and prolonged the duration of the latency period in a dose-dependent manner 4 h after administration. 5. The ED(50) of valproate in the first hour after injection was 63.19 mg/kg (95% confidence interval 51.37-77.71 mg/kg). 6. None of the doses of valproate applied eliminated the EEG signs of metaphit-provoked epileptiform activity. 7. Taken together, these results suggest that all doses of valproate examined acted to suppresse behavioural but not epileptic EEG spiking activity in metaphit-induced seizures.

[Bilobular pancreas: another variant of the divided pancreas?].

BACKGROUND: Pancreas divisum is the most common anomaly of the pancreas. This anomaly has been known as a possible cause of recurrent pancreatitis. CASE REPORT: We performed computerized tomography (CT) of the abdomen in 5 children in whom a divided pancreas was confirmed using endoscopic cholangiopancreatography. In a girl, who had three episodes of severe acute pancreatitis, a CT examination confirmed a completely divided embryonal dorsal and ventral primordium. We named this variant of the divided pancreas the "bilobular pancreas". Contrary to the remaining 4 children in whom the control of the number and severity of attacks, as well as the control of pancreatic pain were achieved by pharmacotherapeutics and an adequate diet, in the reported patient sphincteroplasty of the papilla duodeni minor resulted in a full control of the disease. CONCLUSION: The paper discussed the possibility that the variant of the divided pancreas, with anatomically completely separated ventral and dorsal pancreas and their ductal systems, is the key factor that determines the severity of pancreatic disease and an indication for sphincteroplasty of the papilla duodeni minor as the major therapeutic method.

Delta-sleep-inducing peptide potentiates anticonvulsive activity of valproate against metaphit-provoked audiogenic seizure in rats.

The effect of delta-sleep-inducing peptide (DSIP) on the anticonvulsive activity of a nonprotective valproate (VPA) dose in a metaphit model of generalized, reflex audiogenic seizures in adult Wistar rats was studied. The animals that received metaphit (10 mg/kg) were exposed to audiogenic stimulation (100 +/- 3 dB, 60 s) at hourly intervals. Metaphit-treated rats displaying seizures in 8 previous tests were i.p. injected with VPA (50 mg/kg) or DSIP (1.0 mg/kg) or their combination. Latency to seizure was behaviorally assessed. The EEGs and power spectra were recorded and analyzed. Neurotoxicity was evaluated by the chimney test. DSIP or VPA alone expressed no significant effect on the latency duration, but their combination significantly prolonged latency to seizure during 6 h after injection, while inducing no significant motor impairment. Neither the applied drugs nor their combination abolished metaphit-provoked EEG epileptiform activity. The results show that DSIP potentiated anticonvulsive effects of a nonprotective VPA dose in a metaphit model of audiogenic seizures without influencing its neurotoxicity.

[Clinical manifestations of cow milk protein intolerance in infants].

INTRODUCTION: The disorder of cow milk protein intolerance is characterised by a wide spectrum of clinical manifestations caused by hypersensitivity of type I, II, or IV, and occurs in 2-3% of children, mostly infants. OBJECTIVE: The aim of this study was to present our experiences and observations of clinical signs and symptoms of cow milk protein intolerance in infants aged below 12 months. METHOD: The investigation was carried out on a sample of 55 infants, aged between 1.5-9 months (X = 4.21 +/- 1.25), who had cow milk protein intolerance. Diagnosis of illness was based on characteristic anamnestic, clinical, and laboratory parameters, as well as on an adequate patient's response to antigen elimination. RESULTS: The clinical presentation of cow milk protein intolerance was dominated by cutaneous problems, found in 40/55 (72.73%) infants, followed by digestive disorders, found in 31 (56.36%), while respiratory tract disorders were observed least frequently (14.55%). None of the patients developed anaphylactic shock. Changes involving only one system were found in 35 (63.64%) patients; of these 20 (36.36%) were cutaneous and 15 (27.27%) digestive. Twenty (36.36%) infants displayed multisystemic changes; of these 12 were cutaneus with digestive, 4 were cutaneus with respiratory, while 4 infants had cutaneous, digestive, and respiratory disorders. Of the 55 infants with cow milk protein intolerance, 26 (47.27%) had urticaria, 22 (40.00%) perioral erythema, 21 (38.18%) diarrhoea (15 haemorrhagic, 6 non-haemorrhagic), 13 (23.64%) vomiting, 12 (21.82%) Quincke's oedema, 12 (21.82%) eczema, 5 (9.09%) obstructive bronchitis, while 3 (5.45%) infants had laryngitis. In 5 (9.09%) patients we found a significant body weight deficit and in 3 (5.45%), sideropenic anaemia, while longitudinal growth retardation was not registered in any of the patients. CONCLUSION: Our study showed that the disorder of cow milk protein intolerance predominantly involved cutaneous disorders as well as combinations of cutaneous and digestive disorders, while respiratory system disorders proved to be relatively rare.

[Experimental models of epilepsy]. / Eksperimentalni modeli epilepsije.

INTRODUCTION: An epileptic seizure is a clinical event and epilepsy is rather a group of symptoms than a disease. The main features all epilepsies have in common include, spontaneous occurrence, repetitiveness, and ictal correlation within the EEG. Epilepsies are manifested with distinct EEG changes, requiring exact clinical definition and consequential treatment. Current data show that 1% of the world's population (approximately 50 million people) suffers from epilepsy, with 25% of patients bpeing refractory to therapy and requiring search for new substances in order to decrease EEG and behavioral manifestations of epilepsies. MATERIAL AND METHODS: In regard to discovery and testing of anticonvulsant substances the best results were achieved by implementation of experimental models. Animal models of epilepsy are useful in acquiring basic knowledge regarding pathogenesis, neurotransmitters (glutamate), receptors (NMDA/AIPA/kainate), propagation of epileptic seizures and preclinical assessment of antiepileptics (competitive and non-competitive NMDA antagonists). RESULTS AND CONCLUSIONS: In our lab, we have developed a pharmacologic model of a (metaphit, NMDA and remacemide-cilastatin) generalized, reflex, and audiogenic epilepsy. The model is suitable for testing various anticonvulsant substances (e.g. APH, A4P, CPP, Mk-801) and potential antiepileptics (e.g. DSIP, its tetra- and octaanalogues).

The effects of delta sleep-inducing peptide on incidence and severity in metaphit-induced epilepsy in rats.

The effects of delta sleep-inducing peptide (DSIP) on metaphit- (1-(1(3-isothiocyanatophenyl)-cyclohexyl) (piperidine)-) induced audiogenic seizures in adult male Wistar rats were studied. The animals were divided into four experimental groups: 1. saline injected; 2. metaphit administered (10 mg x kg (-1)); 3. metaphit administered plus DSIP injected (dose range 0.1-1 mg x kg (-1)) and 4. DSIP injected (1 mg x kg (-1)). Upon treatment, the rats were exposed to sound stimulation ( 100 +/- 3 dB, 60 s) at hourly intervals and the incidence and severity (running, clonus and tonus) of seizures were analyzed. In most animals, metaphit led to EEG abnormalities and elicited epileptiform activity recorded as spikes, polyspikes and spike-wave complex and increased power spectra. Time-course studies revealed the peak of convulsive activity 7-12 h after the injection in metaphit-treated rats. DSIP acted as an anticonvulsant and the most potent anticonvulsive dose of 1 mg x kg (-1)significantly increased power spectra of deltawaves (2-11 h) in comparison with the saline-control group and decreased the incidence and duration of convulsive response, as well as mean seizure grade of metaphit-induced convulsions. These results suggest that DSIP should be considered as having potential anticonvulsant activity in this animal model.

[Behavioral and electroencephalographic effects of delta sleep inducing peptide and its analogue on metaphit-induced audiogenic seizures in rats]. / Ponasanje i elektroencefalografska dejstva delta peptida i njegovog analoga u zvucnoj epilepsiji izazvanoj metafitom kod pacova.

INTRODUCTION: Delta sleep inducing peptide (DSIP) is well known natural somnogenic peptide that has many other physiological functions. DSIP analogues representing hepta- and octapeptides (also known as long) as well as tetrapeptide (termed short, used in our experiments) were synthesized with a view to evaluate the peptide specificity in sleep. The effects of DSIP and its analogue DSIP1-4 on metaphit 1-[1(3-isothiocyanatophenylciclohexyl)-piperidine] induced audiogenic seizures were evaluated in rats. METHODS: Male Wistar albino rats were divided into 4 groups: 1. Saline; 2. Metaphit; 3. Metaphit + DSIP, and 4. Metaphit + DSIP1-4. To examine the blocking effects of DSIP and its analogue on fully developed metaphit seizures, the last two groups were injected after the 8th audiogenic testing. Animals were injected with metaphit (10 mg/kg) intraperitoneally (i.p.) and exposed to sound stimulation (100 +/- 3 dB, 60 s) at hourly intervals. The incidence and severity (running, clonus and tonus) of seizures were analyzed. For electroencephalographic (EEG) recordings, three gold-plated electrodes were used. Convulsive behavior was assessed by incidence of motor seizure and by seizure severity grade, determined by descriptive rating scale ranging from 0 to 3: 0--no response, 1--wild running only; 2--wild running followed by clonic seizures of all four limbs with body rollover; 3--wild running progressing to generalized clonic convulsions followed by tonic extension of fore- and hind legs and tail. Sound onset, seizure events, and sound offset, along with the animal's behavior (convulsive or other) were characterized with EEG changes. RESULTS: In most animals, the administration of metaphit resulted in electroencephalographic abnormalities, elicited epileptiform activity in the form of spikes, polyspikes and spike-wave complexes. Maximum incidence and severity of metaphit convulsions occurred 8 h after the injection (9/12, 75%), then abated gradually and disappeared 30 h later. Both DSIP and DSIP1-4 significantly increased the power spectra of d waves and decreased the incidence of seizures, mean seizure grade and tonic component of metaphit-induced convulsions. DISCUSSION: Metaphit has been shown to induce audiogenic seizures after systemic and intracerebroventricular administration and to be truly epileptic in small rodents, although about 8 h after metaphit administration, the power spectra increased and was more intense in the period of sound onset and seizure events. Taken together, DSIP makes an optimal ratio between inhibitory and excitatory amino acid neurotransmitters and may represent one of the endogenous control systems of the brain, thus exerting the protective effect against the seizures. The results obtained throughout the present study corroborate and broaden the data on prolonged antiepileptic DSIP effect. CONCLUSION: The results of the present study strongly suggest that treatment of adult rats with DSIP and its analogue DSIP1-4 should be considered as potential natural antiepileptics.

Induction of audiogenic seizures in imipenem/cilastatin-treated rats.

We investigated the effect of intense audiogenic stimulation (AGS) on rats treated with the antibiotic imipenem and dipeptidase inhibitor cilastatin (Imi/Cil). Under pentobarbital anesthesia (40 mg/kg) adult male Wistar rats were implanted with electrodes and cannulas were placed in the right lateral ventricle. Animals were divided into the following groups: (1) vehicle, (2) Imi/Cil 10 microg/10 microg, (3) Imi/Cil 25 microg/25 microg, (4) vehicle+AGS, (5) Imi/Cil 10 microg/10 microg +AGS, and (6) Imi/Cil 25 microg/25 microg +AGS. Imi/Cil was administered intracerebroventricularly in 5 microl of physiological saline. AGS (100+/-3 dB, 60 seconds) was applied at 15-minute intervals after the injection. Imi/Cil-induced seizures (twitching, forelimb clonus, headnodding, rearing, and clonic convulsions) and Imi/Cil-audio-induced seizures (wild running, clonic and tonic convulsions) were scored according to appropriate rating scales. Imi/Cil provoked convulsions dose-dependently. Each behavioral seizure response had a characteristic EEG correlate. AGS by itself did not provoke seizures in untreated rats. Sound stimulation in Imi/Cil-injected rats elicited typical audiogenic seizures, which were induced during five AGS tests (75 minutes postinjection). In most cases audiogenic seizures were not associated with epileptiform activity in the EEG, indicating that spreading of seizures did not involve the cortex. Since Imi/Cil-induced and Imi/Cil-audio-induced seizures differed behaviorally and electroencephalographically, it is suggested that different neural pathways are responsible for these two types of seizures: neuronal networks in the cortex are involved in Imi/Cil-induced seizures, whereas audiogenic seizures use networks residing primarily in the brainstem.