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Background: Alzheimer’s disease (AD) is a complex neurodegenerative disease with multifactorial etiology, unsatisfactory treatment, and a necessity for broad-spectrum active substances for cure. The mucus from Helix aspersa snail is a mixture of bioactive molecules with antimicrobial, anti-inflammatory, antioxidant, and anti-apoptotic effects. So far there are no data concerning the capacity of snail extract (SE) to affect neurodegenerative disorders. Objective: The effects of SE from Helix aspersa on learning and memory deficits in Alzheimer’s type dementia (ATD) induced by scopolamine (Sco) in male Wistar rats were examined and some mechanisms of action underlying these effects were evaluated. Methods: SE (0.5 mL/100 g) was applied orally through a food tube for 16 consecutive days: 5 days before and 11 days simultaneously with Sco (2 mg/kg, intraperitoneally). At the end of Sco treatment, using behavioral methods, we evaluated memory performance. Additionally, in cortex and hippocampus the acetylcholinesterase (AChE) activity, acetylcholine and monoamines (dopamine, noradrenaline, and serotonin) content, levels of main oxidative stress markers, and expression of brain-derived neurotrophic factor (BDNF) and cAMP response element-binding protein (CREB) were determined. Results: We demonstrated that, according to all behavioral tests used, SE significantly improved the cognitive deficits induced by Sco. Furthermore, SE possessed AChE inhibitory activity, moderate antioxidant properties and the ability to modulate monoamines content in two brain structures. Moreover, multiple SE applications not only restored the depressed by Sco expression of CREB and BDNF, but significantly upregulated it. Conclusion: Summarizing results, we conclude that complex mechanisms underlie the beneficial effects of SE on impaired memory in Alzheimer’s type dementia.

This study evaluates some of the neuromodulatory mechanisms of the memory loss preventive effect of alpha-lipoic acid (ALA) in a scopolamine (Sco)-induced rat model of Alzheimer's disease (AD) type dementia. Our results confirmed that Sco administration induces significant memory impairment, worsens exploratory behaviour and habituation, increases acetylcholinesterase (AChE) activity, and induces pathological monoamine content changes in the prefrontal cortex and hippocampus. ALA administration largely prevented Sco-induced memory impairment. It also improved exploratory behaviour and preserved habituation, and it decreased AChE activity, reversing it to control group levels, and corrected aberrant monoamine levels in the prefrontal cortex and hippocampus. According to the data available, this is the first time that ALA-induced changes in AChE and monoamine levels in the prefrontal cortex and hippocampus (brain structures related to learning and memory) have been demonstrated in a Sco-induced rat model of AD type dementia.

Background: Inhibitors of acetylcholinesterase (AChE) are used to treat many disorders, among which are neurodegenerative upsets, like Alzheimer’s disease (AD). One of the limited licensed AChE inhibitors (AChEIs) used as drugs is the natural compound galantamine (Gal). Objective: As Gal is a toxic compound, here we expose data about its four derivatives in hybrid peptide-norgalantamine molecules, which have shown 100 times lower toxicity. Methods: Four newly synthesized galantamine derivatives have been involved in docking analysis made by Molegro Virtual Docker. Biological assessments were performed on ICR male mice. The change in short and long-term memory performance was evaluated by passive avoidance test. AChE activity and levels of main oxidative stress parameters: lipid peroxidation, total glutathione (GSH), enzyme activities of catalase (CAT), superoxide dismutase, and glutathione peroxidase were measured in brain homogenates. Results: Our experimental data revealed that the new hybrid molecules did not impair memory performance in healthy mice. Two of the compounds demonstrated better than Gal AChE inhibitory activity in the brain. None of them changed the level of lipid peroxidation products, one of the compounds increased GSH levels, and all of them increased CAT enzyme activity. Conclusion: The new galantamine-peptide hybrids demonstrated a potential for inhibition of AChE and antioxidant activity and deserve further attention.

The neuroprotective capacity of newly synthesized amantadine derivative tyrosinyl-amantadine (Tyr-Am) with expected antiparkinsonian properties was evaluated in a 6-hydroxydopamine (6-OHDA) model of Parkinson's disease. Male Wistar rats were divided into the following groups: sham-operated (SO), striatal 6-OHDA-lesioned control group, 6-OHDA-lesioned rats pretreated for 6 days with Tyr-Am (16 mg/kg administered intraperitoneally, i.p.), and 6-OHDA-lesioned rats pretreated for 6 days with amantadine (40 mg/kg i.p.), used as a referent. On the first, second and third week post-lesion, the animals were subjected to some behavioral tests (apomorphine-induced rotation, rotarod, and passive avoidance test). The acetylcholinesterase (AChE) activity and key oxidative stress parameters including lipid peroxidation levels (LPO) and superoxide dismutase (SOD) were measured in brain homogenates. The results showed that the neuroprotective effect of Tyr-Am was comparable to that of amantadine, improving neuromuscular coordination and learning and memory performance even at a 2.5-fold lower dose. Tyr-Am demonstrated significant antioxidant properties via decreased LPO levels but had no effect on AChE activity. We can conclude that the newly synthesized amantadine derivative Tyr-Am demonstrated significant antiparkinsonian activity in a 6-OHDA experimental model.

This study evaluates some of the neuromodulatory mechanisms of the memory loss preventive effect of alpha-lipoic acid (ALA) in a scopolamine (Sco)-induced rat model of an Alzheimer’s disease (AD) type dementia. Our results confirmed that Sco administration induces significant memory impairment, worsens exploratory behaviour and habituation; it increases acetylcholinesterase (AChE) activity and induces pathological monoamine content changes in the brain prefrontal cortex and hippocampus. ALA administration prevented to a large extent Sco-induced memory impairment; it also improved exploratory behaviour and preserved habituation; it decreased AChE activity, reversing it to Control group levels and corrected aberrant monoamine levels in the brain prefrontal cortex and hippocampus. According to the data available, this is the first time that ALA-induced changes in AChE and monoamine levels in the brain prefrontal cortex and hippocampus (brain structures related to learning and memory) have been demonstrated in a Sco-induced rat model of AD type dementia.

Alzheimer’s disease (AD), a progressive neurodegenerative disorder characterized by memory loss and cognitive functions decline, is a leading cause for dementia and currently ranked as the sixth foremost cause of death. As of present, treatment of AD is symptomatic without convincing therapeutic benefits and new, effective, therapeutic agents are pursued. Due to massive loss of cholinergic neurons and decreased acetylcholine levels, cholinesterase inhibitors like galantamine, remain the backbone of pharmacological treatment of the disease. In the present study, using behavioral and biochemical methods, four newly synthesized galantamine derivatives, Gal 34, Gal 43, Gal 44, and Gal 46, were evaluated for a beneficial effect in a scopolamine model of dementia in mice. They were designed to have all the advantages of galantamine and additionally to inhibit β-secretase and exert favorable effects on plasma lipids. Behavioral tests included step-through inhibitory avoidance, T-maze, and the hole-board test, whereas biochemical evaluations involved assessment of acetylcholinesterase activity, brain monoamines levels, lipid peroxidation, catalase, glutathione peroxidase, and superoxide dismutase activities along with measurement of total glutathione. Results show that Gal 43, Gal 44, and, in particular, Gal 46 are especially effective in improving both short- and long-term memory and in the case of Gal 46 having a significant effect on exploratory activity as well. Although Gal 34 did not show behavioral effects as convincing as those of the other three galantamine derivatives, it demonstrated persuasive antioxidant and restorative capacities, making all four galantamine derivatives promising AD treatment agents and prompting further research, especially that in many of our studies they performed better than galantamine.

Previous evidence has shown a link between neurodegenerative diseases, including Parkinson's disease (PD), and melatonin. The data in the literature about the impact of the hormone under different experimental PD conditions are quite controversial, and its effect on memory impairment in the disease is very poorly explored. The current research was aimed at investigating the role of melatonin pretreatment on memory and motor behavior in healthy rats and those with the partial 6-hydroxydopamine (6-OHDA) model of PD. All rodents were pretreated with melatonin (20 mg/kg, intraperitoneally) for 5 days. At 24 h and 7 days after the first treatment for healthy rats, and at the second and third week post-lesion for those with PD, the animals were tested behaviorally (apomorphine-induced rotations, rotarod, and passive avoidance tests). The neurochemical levels of dopamine (DA), acetylcholine (ACh), noradrenaline (NA), and serotonin (Sero) in the brain were also determined. The results showed that in healthy animals, melatonin pretreatment had amnestic and motor-suppressive effects and did not change the levels of measured brain neurotransmitters. In animals with PD, melatonin pretreatment exerted a neuroprotective effect, manifested as a significantly decreased number of apomorphine-induced rotations, reduced number of falls in the rotarod test, and improved memory performance. The brain DA and ACh concentrations in the same animals were restored to the control levels, and those of NA and Sero did not change. Our results demonstrate a beneficial effect of melatonin on memory and motor disturbance in 6-OHDA-lesioned rats.

We compared the neuroprotective action of three natural bio-antioxidants (AOs): ellagic acid (EA), &alpha, lipoic acid (LA), and myrtenal (Myrt) in an experimental model of Parkinson&rsquo, s disease (PD) that was induced in male Wistar rats through an intrastriatal injection of 6-hydroxydopamine (6-OHDA). The animals were divided into five groups: the sham-operated (SO) control group, striatal 6-OHDA-lesioned control group, and three groups of 6-OHDA-lesioned rats pre-treated for five days with EA, LA, and Myrt (50 mg/kg, intraperitoneally- i.p.), respectively. On the 2nd and the 3rd week post lesion, the animals were subjected to several behavioral tests: apomorphine-induced rotation, rotarod, and the passive avoidance test. Biochemical evaluation included assessment of main oxidative stress parameters as well as dopamine (DA) levels in brain homogenates. The results showed that all three test compounds improved learning and memory performance as well as neuromuscular coordination. Biochemical assays showed that all three compounds substantially decreased lipid peroxidation (LPO) levels, and restored catalase (CAT) activity and DA levels that were impaired by the challenge with 6-OHDA. Based on these results, we can conclude that the studied AOs demonstrate properties that are consistent with significant antiparkinsonian effects. The most powerful neuroprotective effect was observed with Myrt, and this work represents the first demonstration of its anti-Parkinsonian impact.

The SARS-CoV-2 virus, first reported in December 2019 in China, is the causative agent of the current COVID-19 pandemic that, at the time of writing (1 November 2020) has infected almost 43 million people and caused the death of more than 1 million people. The spectrum of clinical manifestations observed during COVID-19 infection varies from asymptomatic to critical life-threatening clinical conditions. Emerging evidence shows that COVID-19 affects far more organs than just the respiratory system, including the heart, kidneys, blood vessels, liver, as well as the central nervous system (CNS) and the peripheral nervous system (PNS). It is also becoming clear that the neurological and psychological disturbances that occur during the acute phase of the infection may persist well beyond the recovery. The aim of this review is to propel further this emerging and relevant field of research related to the pathophysiology of neurological manifestation of COVID-19 infection (Neuro-COVID). We will summarize the PNS and CNS symptoms experienced by people with COVID-19 both during infection and in the recovery phase. Diagnostic and pharmacological findings in this field of study are strongly warranted to address the neurological and psychological symptoms of COVID-19.

Close functional and anatomical interactions between the neurotensin (NT) and dopamine (DA) systems suggest that NT could be associated with Parkinson’s Disease (PD). However, clinical use of NT is limited due to its rapid degradation. This has led to the synthesis of a number of new NT fragment 8-13 [NT(8-13)] analogues, such as NT2 and NT4, to avoid the fast biodegradation of NT. The aim of this study was to investigate the neuroprotective effects of NT2 and NT4 on an experimental model of Parkinson’s disease in rats induced with 6-hydroxydopamine (6-OHDA) treatment, producing striatal lesions. Wistar male rats were divided into different groups: a sham-operated (SO) group, a striatal 6-OHDA-lesioned control group, two groups of 6-OHDA-lesioned rats treated for 5 days with NT2 or NT4 (10 mg/kg, intraperitoneally) and a NT-treated group as reference. During the first and second week post lesion the animals were subjected to a number of behavioral tests (apomorphine-induced rotations, rotarod, passive avoidance test), and brain tissue was evaluated histologically and also assessed for DA levels. The results showed that both the number of apomorphine-induced rotations and falls (rotarod test) increased in the 6-OHDA group relative to the SO group. At the same time, the 6-OHDA-treated group showed significant memory impairment, based on the to step-through test, compared to the SO group. Treatment with NT2 and NT4 significantly decreased the number of apomorphine-induced rotations and improved the memory of lesioned animals, with these NT analogues demonstrating better neuroprotective and neuromodulatory effects than NT. DA content in the brain of the PD rats treated with NT2 and NT4 increased, possibly due to attenuation of the loss of DA-ergic neurons. NT2 and NT4 were found to easily penetrate the blood–brain barrier and they showed a better stability than the reference NT neuropeptide. In conclusion, the NT approach represents an attractive strategy for the treatment of neurodegenerative disease.

Pollen and plant macrofossil analyses were performed on a sequence 105 cm deep obtained from a peat bog (1750 m) that is located above the present timber-line in the Osogovo Mountain, Southwestern ...

International audience; Aim: To evaluate the biomization technique for reconstructing past vegetation in the Eastern Mediterranean-Black Sea-Caspian-Corridor using an extensive modern pollen data set and comparing reconstructions to potential vegetation and observed land cover data. Location: The region between 28-48 degrees N and 22-62 degrees E. Methods: We apply the biomization technique to 1,387 modern pollen samples, representing 1,107 entities, to reconstruct the distribution of 13 broad vegetation categories (biomes). We assess the results using estimates of potential natural vegetation from the European Vegetation Map and the Physico-Geographic Atlas of the World. We test whether anthropogenic disturbance affects reconstruction quality using land use information from the Global Land Cover data set. Results: The biomization scheme successfully predicts the broadscale patterns of vegetation across the region, including changes with elevation. The technique discriminates deserts from shrublands, the prevalence of woodlands in moister lowland sites, and the presence of temperate and mixed forests at higher elevations. Quantitative assessment of the reconstructions is less satisfactory: the biome is predicted correctly at 44% of the sites in Europe and 33% of the sites overall. The low success rate is not a reflection of anthropogenic impacts: only 33% of the samples are correctly assigned after the removal of sites in anthropogenically altered environments. Open vegetation is less successfully predicted (33%) than forest types (73%), reflecting the under-representation of herbaceous taxa in pollen assemblages and the impact of long-distance pollen transport into open environments. Samples from small basins (

A palynological study was performed on a 2 m thick sediment sequence recovered from Mire Kupena (1356 m), a former lake in the Western Rhodopes Mountains (south Bulgaria) and supported by radiocarbon dating. The record extends back to ca. 30,000 cal. yrs. BP (Middle Pleniglacial) when the study area was occupied by wooded steppe composed of Pinus sp., Pinus peuce , some Betula , Juniperus , and cold-tolerant herb vegetation dominated by Artemisia and Chenopodiaceae. In addition, the almost continuous presence of deciduous Quercus , Tilia , Corylus , Abies , Picea , Fagus , Alnus , and Carpinus betulus pollen suggests interstadial conditions and this area could be recognized as a montane refugial place. A reconstruction of the interval between ca. 24,000 and 15,000 cal. yrs. BP was not possible due to an extremely low sediment accumulation rates or, more likely, a hiatus. The lateglacial landscape was dominated by mountain-herb steppe vegetation with isolated stands of Pinus , Betula and shrubland of Juniperus . The afforestation in the Early Holocene started with broad-leaved forests composed of Quercus with C. betulus , Carpinus orientalis / Ostrya , Ulmus , Tilia and Corylus and minor amounts of Pinus , Betula and Abies . In the Late Holocene ( Fagus began to gain importance chiefly at the expense of the mixed oak forests, while after ca. 2000 cal. yrs. BP forests of Pinus sylvestris with some P . peuce quickly expanded around the mire. A comparison with other long palynological records from the mountains and lowlands of Bulgaria and in northern Greece reveals not only common trends in the vegetation development that are a reflection of the climate changes, but also site-specific features related to the location and topography of each site.

Atanassova J., Yurukova L., Lazarova M. (2012): Pollen and inorganic characteristics of Bulgarian unifloral honeys. Czech J. Food Sci., 30: 520–526. The melissopalynological characteristics, three main physicochemical parameters (water content, pH, and electrical conductivity), and 19 macro- and microelements contents of 15 honey types from throughout Bulgaria that were col lected from 2006 to 2009 were evaluated. The main honeys studied came from Robinia pseudoacacia L., Helianthus annuus L., Brassica spp., Tilia spp., and Vicia spp. The botanical origins of unifloral honey samples were identified as Lotus spp., Coriandrum sativum L., Daucus-type, Stachys -type , Salix spp., Prunus spp., Castanea sativa Mill ., Paliu rus spina-christi Mill. , Sophora japonica, and Amorpha spp. Based on the physicochemical parameters and elements contents, one sample with high a percentage of Trifolium spp. pollen was identified as honeydew honey.

Vasoactive intestinal peptide (VIP) was administered in a model of zymosan-induced generalized inflammation (ZIGI). Its beneficial action was associated with reduced TNF-α and increased IL-10 production, lowered levels of creatinine, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and bilirubin in circulation. VIP diminished the level of RANTES and MIP-1α in peritoneal exudate and circulation. The neuropeptide inhibited NO release from stimulated peritoneal macrophages. Decreased spleen, liver and kidney enlargement and less pathological changes in liver were observed. The effect of VIP was attenuated by pretreatment with VIP antagonist (anti-VIP) before the induction of shock.

Honey samples collected during 2007-2010 from serpentine and non-serpentine localities in the Eastern Rhodopes Mt. (Bulgaria) were characterized on the basis of their pollen content by qualitative melissopalynological analysis and physicochemical composition. Water content, pH, electrical conductivity, macroelements-K, Ca, Mg, P, and microelements-As, Cd, Co, Cr, Cu, Fe, Mn, Na, Ni, Pb, and Zn were determined after the Harmonised Methods of the International Honey Commission and ICP-AES method. The results from serpentine honey samples were compared with data from bee pollen collected from the same serpentine area. Different elements have different concentrations in honey from the same botanical type even collected from the same geographical region, same locality, and same beehive but in different vegetation season. The elements Mg, Mn, Ni, and P contribute mostly for separation of the serpentine honey samples based on measured elemental concentrations and performed principal component analysis. The element concentrations were higher in bee pollen and above the permissible limits for the toxic metals Cd and Pb. No specific indicator plant species was found for identification of the geographical origin of serpentine honey in relation to the forage of bees.

The Late Glacial and early-Holocene vegetational history of a newly dated pollen and macrofossil diagram from Besbog, a cirque lake at 2250 m just above the forest limit in the Pirin Mountains of southwestern Bulgaria, is compared with a newly dated pollen diagram for the mire Shiroka Polyana at 1400 m in the conifer forest of the nearby Rhodope Mountains in order to investigate the chronology of major changes in the vegetation at different elevations. In the Lake Besbog record the non-arboreal pollen assemblage of the Late Glacial changed abruptly to that of Betula, Quercus and other deciduous types. The date for this change is about 11.6 ka cal b.p. The Quercus assemblage may be composed of pollen blown from intermediate elevations, to which deciduous forest had expanded because of higher summer temperatures related to high summer insolation. At Shiroka Polyana (1400 m) in the modern conifer belt, a similar change did not occur until about 8.8 ka cal b.p. The persistence of the dry steppe or steppe forest in the early Holocene at this lower site can also be attributed to high summer insolation. Thus as atmospheric temperature increased at the end of the Late Glacial, deciduous forests expanded first at intermediate elevations in the Pirin Mountains and only later in the Rhodope Mountains at lower elevations as summer insolation decreased.

Site details Mire Kupena is a former lake (41◦ 59′ 07.5′′ N, 24◦ 19′ 05.1′′ E; 1356 m above sea level (a.s.l.)) located on the territory of the Biosphere Reserve Kupena, Western Rhodopes Mountains, Bulgaria. The reserve is located between 600 and 1400 m a.s.l. on a north-facing slope and its vegetation is rather diverse. In the lower parts, the most common plant communities are those of Quercus dalechampii Ten. mixed in some places with Carpinus betulus L., Fagus sylvatica L., Pinus nigra Arn. and Pinus sylvestris L. At higher altitudes, plant communities of F. sylvatica dominate, followed by those of Pinus sylvestris with some Abies alba Mill. and Picea abies (L.) Karst. The mire is formed in a depression on an area of 6 ha, surrounded by an almost pure forest of Pinus sylvestris with an admixture of Picea abies, A. alba, F. sylvatica, Betula pendula Roth., Sorbus aucuparia L., Q. dalechampii and an undergrowth of Juniperus communis L., Vaccinium myrtillus L. and V. vitis-idaea L. The mire vegetation is represented by sparse moss cover (Sphagnum spp.) and includes various species of Carex, Juncus, Potentilla, Ranunculus, Galium. The basin is fed by rainfall and water from melting snow. During the last two decades most of the mire surface on the hummocks has been overgrown by pine trees (Huttunen et al., 1992; Bozilova et al., 2011). The climate in the area is montane with a mean annual temperature of 5–10 ◦S and a mean annual precipitation of 600–800 mm with a maximum in May–June. The basic soil types are brown forest (60%), humic-carbonate and cinnamomic-forest.

The modulation of striatal cholinergic neurons by somatostatin (SOM) was studied by measuring the release of acetylcholine (ACh) in the striatum of freely moving rats. The samples were collected via a transversal microdialysis probe. ACh level in the dialysate was measured by the high performance liquid chromatography method with an electrochemical detector. Local administration of SOM (0.1, 0.5 and 1 microM) produced a long-lasting and concentration-dependent increase in the basal striatal ACh output. The stimulant effect of SOM was antagonized by the SOM receptor antagonist cyclo(7-aminopentanoyl-Phe-D-Trp-Lys-Thr[BZL]) (1 microM). In a series of experiments, we studied the effect of 6,7-dinitroquinoxaline-2, 3-dione (DNQX), a selective non-NMDA (N-methyl-D-aspartate) glutamatergic antagonist, on the basal and SOM-induced ACh release from the striatum. DNQX, 2 microM, perfused through the striatum had no effect on the basal ACh output but inhibited the SOM (1 microM)-induced ACh release. The non-NMDA glutamatergic receptor antagonist 1-(4-aminophenyl)-4-methyl-7,8-methylendioxy-5H-2,3- benzodiazepine (GYKI-52466), 10 microM, antagonized the SOM (1 microM)-induced release of ACh in the striatum. Local administration of the NMDA glutamatergic receptor antagonist, 2-amino-5-phosphonopentanoic acid (APV), 100 microM, blocked SOM (1 microM)-evoked ACh release. Local infusion of tetrodotoxin (1 microM) decreased the basal release of ACh and abolished the 1 microM SOM-induced increase in ACh output suggesting that the stimulated release of ACh depends on neuronal firing. The present results are the first to demonstrate a neuromodulatory role of SOM in the regulation of cholinergic neuronal activity of the striatum of freely moving rats. The potentiating effect of SOM on ACh release in the striatum is mediated (i) by SOM receptor located on glutamatergic nerve terminals, and (ii) by NMDA and non-NMDA glutamatergic receptors located on dendrites of cholinergic interneurones of the striatum.

ispartof: Grana: an International Journal of Palynology vol:50 issue:2 pages:162-164 status: published

The results of pollen analysis of two sediment cores from lake Srebarna (northeast Bulgaria) are presented. On the basis of the palynological data and the radiocarbon dates a reconstruction of the past flora and vegetation is made. For the first time a continuous palaeosuccession is established for the area of the Danubian Dobrudza and the Ludogorie district of northeast Bulgaria: a dominance of mixed xero- and mesophilous oak woods with Carpinus betulus, Ulmus, Tilia, Corylus during the Atlantic period; mixed oak woods with increasing importance of Tilia, Ulmus, Acer and a considerable presence of Carpinus betulus and Fagus during the Sub-boreal period; degradation of the woodland vegetation and transition to secondary plant communities with Carpinus orientalis and herbs during the Sub-atlantic period.

The peat-bog Begbunar (42°09¢ N, 22° 33¢ E; 1750 m a.s.l.) is located in the central treeless zone of the Osogovo mountains, which are situated at the border between south-western Bulgaria and the north-eastern former Yugoslav Republic of Macedonia. This bog was formed near a freshwater spring on a north-western slope, which continues into a steep, deep ravine where isolated stands of beech (Fagus sylvatica L.) exist. The modern mountain vegetation consists of several vegetation belts: the oak-hornbeam belt (up to 1000 m), the compact beech belt (1000 – 1900 m) with fragments of conifers (Pinus nigra J. F. Arnold and Abies alba Mill.) and the subalpine belt, which is occupied by plant communities of Juniperus sibirica Burgsd., Vaccinium myrtillus L., Bruckenthalia spiculifolia (Salisb.) Rchb., Chamaecytisus absinthioides Janka (Kuzm.), Nardus stricta L., and scattered groups of Pinus sylvestris L., etc. The present-day vegetation composition has been strongly influenced by the long-lasting anthropogenic impact, including ore-mining industry and erosion caused by deforestation (Velcev & Tonkov, 1986; Tonkov, 2003). Geologically, the massif is composed mainly of Palaeozoic metamorphic and intrusive rocks, while geomorphological evidence suggests that small valley glaciers existed in the highest parts during Quaternary glaciations. The climate above 1000 m is typical montane with a mean annual precipitation of 700–900 mm. The basic soil types are cinnamomic-forest, brown forest and mountainous-meadow (Velcev et al., 1994).

3-Aminocycloalkanespiro-5-hydantoins were synthesized and their biological activity was studied. In contrast to hydantoins, these compounds failed to induce either anticonvulsive effects in the central nervous system or inhibitory effects on cholinergic contractions in the enteric nervous system. However, they exerted well pronounced, atropinsensitive, contractile effects on the guinea-pig ileum longitudinal muscle preparations. Structure–activity relationships established allow the assumption that: (i) the reduction of the ring size in the molecule of the spirohydantoins leads to an increase in the potency of the respective analogue to induce contractile effect; (ii) the introduction of NH 2 in position 3 increases the ability of all the compounds studied to exert contractions; (iii) the enlargement of the ring leads to: (1) an increase of the degree of desensitization of the preparations; and (2) a decrease (except 1a ) of the potency of the analogues to exert contractile effects.

The effects of the non-N-methyl-D-aspartate (NMDA) agonist quisqualate (QUIS) and selective AMPA/kainate receptor antagonist 1-(aminophenyl)-methyl-7, 8-methyilendioxy-5H-2,3-benzodiazepine (GYKI 52466) on the release of acetylcholine (ACh) from the hippocampus and striatum of freely moving rats were studied by transversal microdialysis. Acetylcholine level in the dialisate was measured by the high performance liquid chromatography (HPLC) method with an electrochemical detector. The QUIS (100 microM) perfused through the striatum induced an increase of extracellular ACh level (250%) which lasted for over 1h and gradually returned to basal values. Local perfusion of GYKI 52466 (10-100 microM) to the striatum did not change the basal release of ACh. GYKI 52466 (10 microM) administered together with QUIS (100 microM) in he striatum antagonized the stimulant effect of QUIS on the ACh release. Local administration of the QUIS (100 microM) through the microdialysis fiber implanted in the hippocampus, caused a long lasting increase of extracellular hippocampal ACh level (360%) which was reversed when the drug was withdrawn from the perfusion solution. The stimulant effect of QUIS was antagonized by concomitant perfusion of GYKI (10 microM). No effect was seen on the basal ACh release when GYKI (10-100 microM) was perfused through the hippocampus. Local perfusion with tetrodotoxin (1 microM) decrease the basal release of ACh and prevented the QUIS-induced increase of ACh both in the hippocampus and striatum. Our in vivo neurochemical results indicate that hippocampal and striatal cholinergic systems are regulated by non-NMDA (probably AMPA) glutamatergic receptors located in the hippocampus and striatum.