Cannabis roots: Pharmacological and toxicological studies in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: There are many studies and therapeutic properties attributed to the flowers and leaves of the Cannabis species, but even with few pharmacological studies, Cannabis sativa L. (Cannabaceae) roots presents several therapeutic indications in folk medicine. AIM OF THE STUDY: This study aimed to evaluate the anti-inflammatory and spasmolytic effects as well as the toxicological profile of the aqueous extract of Cannabis sativa roots (CsAqEx) in mice. MATERIALS AND METHODS: We assessed the anti-inflammatory effect with carrageenan-induced leukocyte migration assay, and carrageenan and histamine-induced paw edema methods; The spasmolytic effect was assessed through in vitro assays with isolated mice trachea. To assess motor coordination and mobility, mice went through the rotarod and open field tests, respectively. For the single-dose toxicity study, we administered CsAqEx at the dose of 1000 mg/kg by gavage. In a repeated dose toxicity study, animals received CsAqEx at doses of 25 mg or 100 mg/kg for 28 days. RESULTS: The CsAqEx inhibited the migration of leukocytes at the doses of 25, 50, and 100 mg/kg. The CsAqEx showed anti-inflammatory activity after the intraplantar injection of carrageenan, presenting a reduction in edema formation at all tested doses (12.5, 25, 50 and 100 mg/kg). The dose of 12.5 mg/kg of CsAqEx prevented edema formation after intraplantar injection of histamine. In an organ bath, 729 µg/mL of CsAqEx did not promote spasmolytic effect on isolated mice tracheal rings contracted by carbachol (CCh) or potassium chloride (KCl). We did not observe clinical signs of toxicity in the animals after acute treatment with CsAqEx, which suggested that the median lethal dose (LD50) is greater than 1000 mg/kg. Repeated dose exposure to the CsAqEx did not produce significant changes in hematological, biochemical, or organ histology parameters. CONCLUSIONS: The results suggest that the anti-inflammatory effect of CsAqEx is related to the reduction of vascular extravasation and migration of inflammatory cells, without effects on the central nervous system. Moreover, there was no spasmolytic effect on airway smooth muscle and no toxicity was observed on mice.

Pharmacological investigation of activities pertaining to modulation of gastrointestinal, respiratory and cardiovascular parameters by Indigofera argentea in experimental models.

Indigofera argentea is widely used for the management of gastrointestinal, respiratory and cardiac disorders. This study was done to explore scientific basis of its uses. Aqueous methanolic extract of Indigofera argentea and its fractions were studied on isolated tissues of rabbit's jejunum, trachea, aorta and atrium. Castor oil induced diarrheal model was used for the study of the antidiarrheal effect and pre-anesthetized rats were used for hypotensive study. Concentration dependent spasmolytic effect of the extract upon isolated jejunum, trachea and aorta was observed. Concentration response curves constructed upon isolated rabbit jejunum, revealed the presence of calcium channel blocker in the plant extract. Moreover, significant reduction (P<0.05) in atrial force of contraction but non-significant reduction in rate of contraction was seen by the application of plant extract. Protection (P<0.05) against diarrhea was observed by the administration of crude extract to rats which were pretreated with castor oil. When given to rats intravenously, the extract showed hypotensive effect. Experimental findings justified the traditional uses of Indigofera argentea on pharmacological basis for the management of disorders pertaining to gut, airway and hypertensive situation.

Antidiarrheal activity of methanol extract of Sophora tonkinensis in mice and spasmolytic effect on smooth muscle contraction of isolated jejunum in rabbits.

Context: In China, the herb Sophora tonkinensis Gagnep. (Fabaceae, ST) (Committee of National Pharmacopeia. 2015) exhibits anti-inflammatory, antitumor, and antiviral effects. However, to date, there have been few studies on its gastrointestinal effect. Objective: The gastrointestinal effect of the methanol extract of ST rhizome (STR) was evaluated. Materials and methods: Study was conducted from February to December 2018. In vivo, antidiarrheal activity of STR (125, 250 and 500 mg/kg; orally) in castor oil-induced diarrheal mice was studied. In vitro, the effects of STR (0.01-10 mg/mL) on the isolated tissue preparations of rabbit jejunum were also investigated, the rabbit jejunum stripes were pre-contracted with Ach (10-5 M), K+ (60 mM) and tested in the presence of STR, the possible spasmolytic effect was analyzed in the pretreatment of the jejunum preparations with STR or verapamil in Ca2+-free high-K+ (60 mM) solution containing EDTA. Results: STR (125, 250 and 500 mg/kg) exhibited antidiarrheal activity. STR (0.01-10 mg/mL) completely relaxed spontaneously contracting, Ach (10-5 M) and high K+ (60 mM) induced contracted jejunum with an EC50 value of 0.66 (0.49-0.96), 0.39 (0.28-0.44) and 0.17 (0.10-0.21), similar to verapamil. Concentration-response curves of CaCl2 could be significantly moved to the right and down in the presence of STR (0.3, 1 mg/mL). Discussion and conclusions: Results suggest the presence of antidiarrheal activity and spasmolytic effects of STR, possibly mediated through Ca2+ channel blocking activity, providing the pharmacological basis for its traditional uses in gastrointestinal disorders.

The impact of proximol (Cymbopogon proximus) intake on pregnant albino ratsand their fetuses during gestation period / Impacto de la ingesta de proximol (Cymbopogon proximus) en las ratas albino embarazadas y sus fetos durante el período de gestación

Halphabarol, the active principle of Proximol, is the most potent of the four antispasmodics present in the national desert weed Cymbopogon proximus or ''Halfa Bar''. Halphabarol is of great value for the management of renal colic and in the expulsion of ureteric calculi as it causes dilation of the ureter below the site of calculus while active propulsion is maintained. Evaluation the congenital malformation of proximol in pregnant albino rats during gestation period. The virgin female rats were mated with male rats and the pregnant rats were orally administered a human equivalent dose (0.05 mg/kg) of Proximol from 5th-20th gestation day. At day 20 of pregnancy, all rats were anesthetized to obtained maternal and fetal data. The treatment group displayed some disorders, which can be summarized as growth retardation, external anomalies, embryonic resorption, and skeletal malformation. We concluded that the oral administration of Proximol resulted in embryonic abnormalities and skeletal malformations.

Halphabarol, el principio activo de Proximol, es el más potente de los cuatro antiespasmódicos presentes en la maleza desértica nacional "Cymbopogon proximus" o "Halfa Bar". Halphabarol es de gran utilidad para el manejo de cólicos renales y para la expulsión de cálculos ureterales, ya que causa la dilatación del uréter por debajo del sitio de cálculo mientras se mantiene el mecanismo de propulsión activa. Se realizó una evaluación de la malformación congénita por Proximol en ratas albinas gestantes durante el período de gestación. Las ratas fueron apareadas y a las ratas gestantes se les administró oralmente, del 5 al 20 día de gestación, una dosis de Proximol (0,05 mg / kg), equivalente a la dosis humana. Al día 20 de gestación, todas las ratas fueron anestesiadas para obtener datos maternos y fetales. El grupo de tratamiento mostró algunos trastornos, que pueden resumirse como retraso del crecimiento, anomalías externas, resorción embrionaria y malformación esquelética. Concluimos que la administración oral de Proximol resultó en anomalías embrionarias y malformaciones esqueléticas.

[The chemical toxicological study of cyclomed].

The objective of the present study was to develop the methods for the chemical and toxicological analysis of cyclopentolate contained in the material evidence for the purpose of forensic chemical and forensic toxicological expertises. The optimal conditions for the isolation of cyclopentolate from the cyclomed preparation and biological fluids were created using a chloroform-2-ptropanol mixture at pH of the medium 10.0. The substance of interest was identified with the use of the color and flocculation reactions, UV spectroscopy, thin layer chromatography and gas chromatography with the use of a mass-selective detector. HPLC and GCh/MS were employed for the quantitative determination. The results of the two methods are in excellent agreement.

A review of the relaxant effect of various medicinal plants on tracheal smooth muscle, their possible mechanism(s) and potency.

ETHNOPHARMACOLOGICAL RELEVANCE: The therapeutic effects of the medicinal plants described in the current review on obstructive pulmonary diseases have found mention in ancient Iranian medical texts and in traditional folk medicine. These effects are attributed to their bronchodilatory activity, which relaxes the smooth muscles of the airway. Therefore, in the present review, the relaxant effects of various extracts, fractions and constituents of medicinal plants on tracheal smooth muscle are reviewed in light of their therapeutic effects on obstructive pulmonary diseases. MATERIALS AND METHODS: The online literature was searched using Medline, PubMed, ScienceDirect, Scopus, Google Scholar, Web of Science and SID (for articles written in Persian). Moreover, local books on ethnopharmacology from 1918 to 2014 were searched with keywords such as tracheal smooth muscle, airway smooth muscle, relaxant effect, bronchodilatory effect and related mechanisms to identify studies on the relaxant effects of medicinal plants on tracheal smooth muscle and the possible mechanism(s) of these effects. RESULTS: All studied plants showed significant relaxant effects on tracheal smooth muscle, which were similar or superior to the effect of theophylline at the used concentrations. According to the results, most of these plants also showed an inhibitory effect on muscarinic and histamine (H1) receptors, whereas some plants showed more pronounced stimulatory effects on the beta-adrenergic receptor. Some of the studied plants also showed inhibitory effects on calcium and potassium channels. CONCLUSION: The present article reviewed the relaxant effects of several medicinal plants on tracheal smooth muscle, which were comparable or superior to the effect of theophylline at the studied concentration. The possible mechanisms of the relaxant effects of the studied medicinal plants and a comparison of these effects were also reviewed. This review presents the fractions and constituents of plants with potent relaxant effects on tracheal smooth muscle, which can be used to treat obstructive pulmonary disease.

In vitro relaxant and spasmolytic effects of essential oil of Pistacia integerrima Stewart ex Brandis Galls.

ETHNOPHARMACOLOGICAL RELEVANCE: Pistacia integerrima J.L. Stewart ex Brandis (Family: Anacardiaceae) galls are used in Indian ethnomedicine for its anti-asthmatic, sedative and spasmolytic properties, however, there are no scientific studies demonstrating its spasmolytic activity. The present investigation deals with the evaluation of relaxant and spasmolytic activities of the essential oil isolated from the galls of Pistacia integerrima J.L. Stewart ex Brandis (EOPI). MATERIALS AND METHODS: In vitro pharmacological assays were carried out on rabbit jejunum spontaneous contractions, guinea pig ileum. The present investigation studied the relaxation of basal tone of isolated guinea pig ileum by possible involvement of NO, prostaglandins, membrane Na(+) channels, potassium channel, enteric nervous system, adrenoceptors, Ca(2+) channels. Additional studies were conducted for comparison of the relaxant effects of EOPI on CaCl2 induced contraction in calcium free tyrode solution, effect on nifedipine insensitive component of ACh-induced contraction and on the contractile machinery to intracellular [Ca(2+)] on isolated guinea pig ileum. RESULTS: EOPI at non-relaxing dose potentiated the isoprenaline induced relaxation of rabbit jejunum. EOPI (50 µg/mL) exhibited 28% relaxation of basal tone of 60 mM K(+) induced contraction which is unaltered by preincubation with 0.5 mM hexamethonium, 0.5 µM Tetrodotoxin, 1 µM indomethacin, and 100 µM L-NG-Nitroarginine Methyl Ester (L-NAME). EOPI inhibited Ca(2+) induced contraction of isolated guinea pig ileum in Ca(2+) free medium. EOPI (10 µg/ml) potentiated the reversal of a KCl-induced tonic contraction has been observed in Ca(2+) free medium. CONCLUSION: The present investigation reinforces the use of Pistacia integerrima Stewart ex Brandis as antispasmodic in folk medicine. Moreover, it is demonstrated the involvement of ß- adrenoceptors and calcium channels in this activity, but not the participation of nicotinic receptors, Na(+) channels, prostaglandins or nitric oxide.

Constipation enhances the propensity to seizure in pentylenetetrazole-induced seizure models of mice.

Epilepsy is characterized by spontaneous recurrent seizures and represents one of the most frequent neurological diseases, affecting about 60 million people worldwide. The cellular and neurocircuit bases of epilepsy are poorly understood. Constipation is a common gastrointestinal disorder characterized by symptoms such as straining, hard stool, and infrequent defecation. Population-based studies have shown that the prevalence of constipation is up to 30% of the population in developed countries. The causal link between seizure and constipation is a common belief among patients and physicians, but there are no scientific data to support this association. The current investigation evaluated the effects of constipation induced by loperamide (a peripheral µ-opioid receptor agonist without effect on central nervous system receptors) and clidinium (a quaternary amine antimuscarinic agent with reduced central nervous system effects) on two different seizure models of mice: (1) myoclonic, clonic, and generalized tonic seizures and death induced by intraperitoneal administration of pentylenetetrazole and (2) clonic seizure threshold induced by intravenous infusion of pentylenetetrazole. We demonstrated that the measured intestinal transit (%intestinal transit) decreased after loperamide or clidinium treatment for 3days. Constipation in mice which was induced by loperamide or clonidine caused a decrease in threshold to clonic seizure in the intravenous pentylenetetrazole seizure model. Moreover loperamide- or clidinium-induced constipation decreased latencies to, clonic, and tonic seizures and death in the intraperitoneal pentylenetetrazole model of mice. Serum ammonia levels were slightly elevated in both loperamide- and clidinium-treated mice. In conclusion, loperamide- or clidinium-induced constipated mice are more prone to seizure which might confirm the belief of patients and physicians about constipation as a trigger of seizure.

NK1 receptors antagonism of dorsal hippocampus counteract the anxiogenic-like effects induced by pilocarpine in non-convulsive Wistar rats.

Recent evidence supports a role for the substance P (SP) in the control of anxiety and epilepsy disorders. Aversive stimuli alter SP levels and SP immunoreactivity in limbic regions, suggesting that changes in SP-NK1 receptor signaling may modulate the neuronal excitability involved in seizures and anxiogenesis. The involvement of NK1 receptors of the dorsal hippocampus and lateral septum in the anxiogenic-like effects induced by a single injection of pilocarpine (PILO) was examined in non-convulsive rats evaluated in the elevated plus-maze (EPM). Male Wistar rats were systemically injected with methyl-scopolamine (1mg/kg) followed 30 min later by saline or PILO (350 mg/kg) and only rats that did not present status epilepticus were used. One month later, vehicle or FK888 (100 pmol) - an NK1 receptor antagonist - were infused in the dorsal hippocampus or the lateral septum of the rats and then behaviorally evaluated in the EPM. Previous treatment with PILO decreased the time spent in and the frequency of entries in the open arms of the EPM, besides altering risk-assessment behaviors such as the number of unprotected head-dipping, protected stretch-attend postures and the frequency of open-arms end activity, showing thus a long-lasting anxiogenic-like profile. FK888 did not show any effect per se but inhibited the anxiogenic responses induced by PILO when injected into the dorsal hippocampus, but not into the lateral septum. Our data suggest that SP-NK1 receptor signaling of the dorsal hippocampus is involved in the anxiogenic-like profile induced by PILO in rats evaluated in the EPM test.

Brine shrimp cytotoxicity of crude methanol extract and antispasmodic activity of α-amyrin acetate from Tylophora hirsuta wall.

BACKGROUND: We have previously reported that aerial parts of Tylophora hirsuta have antispasmodic profile. The current work is an attempt for isolation of pharmacologically active compound(s) that contribute for its antispasmodic activity. METHODS: Preliminary phytochemical screening for crude methanol extract of Tylophora hirsuta (Th.Cr) is performed. Brine shrimp cytotoxicity of crude methanol extract is performed. Column chromatography was used for isolation of compounds. Mass spectroscopy, H(1) NMR and C(13) NMR were used for structural determination of compounds. α-amyrin acetate was tried for possible spasmolytic activity in rabbit's jejunal preparations and KCl-induced contractions. RESULTS: Th.Cr tested positive for saponins, alkaloids, flavonoids and terpenoids. Compound 1 was isolated as α-amyrin acetate. Compound 2 was heptaeicosanol. Crude methanol extract tested positive for brine shrimp cytotoxicity with LC(50) 492.33± 8.08 mg/ml. Compound 1 tested positive for antispasmodic activity on spontaneous rabbits' jejunum preparations with EC(50) (60 ± 2) × 10(-5)M. The compound also tested positive on KCl induced contractions with EC(50) (72 ± 3) × 10(-5)M. CONCLUSIONS: The present work confirms that α-amyrin acetate is has antispasmodic profile and the relaxant effect may be attributed to α-amyrin acetate which is a major compound.

Acute toxicity, brine shrimp cytotoxicity and relaxant activity of fruits of callistemon citrinus curtis.

BACKGROUND: Callistemon citrinus Curtis belongs to family Myrtaceae that has a great medicinal importance. In our previous work, fruits of Callistemon citrinus were reported to have relaxant (antispasmodic) activity. The current work describes the screening of fractions of the crude methanol extract for tracing spasmolytic constituents so that it shall help us for isolation of bioactive compounds. Acute toxicity and brine shrimp cytotoxicity of crude methanol extract are also performed to standardize it. METHODS: The crude methanol extract was obtained by maceration with distilled water (500 ml) three times and fractionated successively with n-hexane, chloroform, ethyl acetate and n-butanol (300 ml of each solvent). Phytochemical analysis for crude methanol extract was performed. Acute toxicity studies were performed in mice. Brine shrimp cytotoxicity studies were performed to determine its cytotoxicity and standardize it. In other series of experiments, rabbits' jejunum preparations were used in screening for possible relaxant activities of various fractions. They were applied in concentrations of 0.01, 0.03, 0.1, 0.3, 1.0, 3.0, 5.0 and 10.0 mg/ml on spontaneous rabbits' jejunum preparations. In similar fashion, fractions were also tested on KCl (80 mM) -induced contractions. Calcium chloride curves were constructed in K-rich Tyrode's solution. The effects of various fractions were tested on calcium chloride curves at concentrations 1.0, 3.0, 5.0 and 10.0 mg/ml. Curves of verapamil used as reference drug at concentration 0.1 µM and 0.3 µM were also constructed. The curves were compared with their respective controls for possible right shift. RESULTS: Methanol extract tested strongly positive for saponins and tannins. However, it tested mild positive for presence of proteins, amino acids, carbohydrates and phenolic compounds. LD(50) value for crude methanol extract is 476.25 ± 10.3 (470-481, n = 4) mg/ml. Similarly, EC(50) value for brine shrimp cytotoxicity is 65.5 ± 7.28 (60.8- 69.4, n = 4) mg/ml. All the fractions relaxed the spontaneous and KCl-induced contractions. EC(50) values (mg/ml) for effects of ethyl acetate fraction on spontaneous and KCl induced contractions are 2.62 ± 0.78 (2.15-3.0, n = 4) and 3.72 ± 0.86 (3.38-4.28, n = 4) respectively. Respective EC(50) values (mg/ml) for n-butanol fraction are 3.59 ± 0.2(3.07-3.9, n = 4) for spontaneous, and 5.57 ± 0.2 (5.07-6.11, n = 4) for KCl- induced contractions. EC(50) value for control calcium chloride curve (without extract) is -2.73 ± 0.19 (-2.6 - -2.81, n = 4) while EC(50) for curves treated with 5.0 mg/ml of chloroform is -2.22 ± 0.02 (-2.16 - -2.3, n = 4). EC(50) value for ethyl acetate treated (1.0 mg/ml) tissues is -1.95 ± 0.10 (-1.88 - -2.0, n = 4) vs. control EC(50) = -2.71 ± 0.08 (-2.66 - -2.76, n = 4). All the fractions, except n-hexane, showed a right shift like that of verapamil (EC(50) = -1.72 ± 0.15 (-1.62 - -1.8, n = 4) vs. Control EC(50) = -2.41 ± 0.06 (-2.38 - - 2.44, n = 4), a standard drug that blocks voltage operated calcium channels. CONCLUSION: Relaxant constituents were more concentrated in ethylacetate fraction followed by chloroform, n -butanol and aqueous fractions that warrant for its isolation. The crude methanol extract is safe at concentration 250 mg/ml or below and results of brine shrimp cytotoxicity assay imply the plant specie may be a source of cytotoxic agents.

Diazepam protects against the enhanced toxicity of cocaine adulterated with atropine.

We examined the toxicity of cocatropine (cocaine/atropine mixture) and the therapeutic potential of diazepam on some behavioral and physiological parameters in rats. Atropine (20 and 60 mg/kg) or cocaine (40 mg/kg) alone did not induce any seizure or death, but the combination significantly increased both, after both acute and binge treatment. There was a significant increase of EEG mean total spectral power in cocatropine- in comparison with cocaine-treated animals. Hyperlocomotion was observed in non-seizuring rats treated with cocaine or cocatropine. Cocaine, atropine 60, and cocatropine (40 + 20 and 40 + 60) all induced hyperthermic effects in non-seizuring rats, while cocatropine (40 + 60)-seizuring animals had hypothermia. An initial hypertensive and tachycardiac effect within 15 min was followed by a secondary fall in the cocatropine (40 + 60) group. Cocatropine toxicity was partially or fully reversed by diazepam (5 mg/kg), given intraperitoneally after the first seizure. The present findings provide, for the first time, details of a synergistic toxic effect of the cocaine/atropine mixture and of the potential of diazepam for treating cocatropine-related hospital emergencies.

Swallowing disorder and inhibition of cough reflex induced by atropine sulfate in conscious dogs.

In this study, the effects of atropine sulfate (atropine) on swallowing and cough reflex were evaluated in the two experimental models in conscious dogs. To evaluate the effects of atropine on swallowing, 1 mL of marker (contrast medium) was injected into the pharynx under X-ray exposure to induce swallowing. Baclofen, used as a positive control, caused marker congestion in the upper esophagus. In our experimental model, atropine (0.02 and 0.1 mg/kg, i.v.) dose-dependently increased not only the number of marker congestions but also that of the swallows. In addition, atropine significantly shortened the onset of first swallowing. In the evaluation of atropine effects on electrically evoked cough reflex induced by two electrodes implanted into the trachea, atropine strongly inhibited the number of coughs at 0.01 or 0.05 mg/kg accompanied with 0.01 or 0.05 mg/kg per hour (i.v.), respectively. These findings indicate that atropine has the potential of causing aspiration pneumonia through induction of swallowing disorder and inhibition of the cough reflex.

Mikania laevigata syrup does not induce side effects on reproductive system of male Wistar rats.

Mikania laevigata (Asteraceae) is a native plant from South America and popularly used as antispasmodic and to treat respiratory diseases. Coumarin is the major chemical substance found in this plant, which have been shown to have antifertility activity in female rats. This study evaluates the toxicity of the exposure to the Mikania laevigata syrup using coumarin as chemical marker on reproductive endpoints in male Wistar rats. Endpoints including reproductive organs weight, sperm and spermatids numbers and sperm morphology were evaluated. Animals were treated daily with Mikania laevigata syrup (3.5; 7.0 and 14.0mg/kg of coumarin) during 90 days by oral gavage. No alterations were observed in body and organ weights, sperm and spermatids numbers as well as sperm morphology of the male rats after the exposure to the Mikania laevigata syrup. Results therefore suggest absence of male reproductive toxicity of the Mikania laevigata syrup at tested doses.

Analgesic and antispasmodic activities of 2-(2-nitro-phenyl)-1H-benzimidazole 5-carboxylic acid: evidence for the importance of the 2-(o-substituted phenyl) group.

Benzimidazole 5-carboxylic acid derivatives were investigated for analgesic activity in this study. Of the benzimidazole compounds tested, 2-(2-nitro-phenyl)-1H-benzimidazole 5-carboxylic acid showed remarkable naloxone sensitive analgesic activity in the tail clamp but not in the tail immersion analgesia tests. This centrally active compound showed antispasmodic activity only on KCl induced contractions of isolated rat ileum and not on acetylcholine induced contractions. Acute toxicity of the compounds were >100 mg/kg i.p. mice. It was concluded that substitution of the 2(o-phenyl) by nitro- but not by chloro- or methoxy groups is important for naloxone sensititive analgesic activity of benzimidazole compounds and it was hypothetized that new imidazole compounds having a 2-(o-substituted phenyl) moiety needs to be investigated.

New insights into the pharmacological properties of potent antiplatelet 2-amino-benzo[d]isothiazol-3-one derivatives.

A series of 2-amino-benzo[d]isothiazol-3-one derivatives (1-8), previously described as in vitro potent antiaggregatory agents endowed with spasmolytic properties, was evaluated for in vitro antiplatelet activity in guinea-pig platelet rich plasma and for in vivo effects on experimental thrombosis and bleeding time in mice. All the 2-amino-benzo[d]isothiazol-3-one derivatives 1-8 were more potent antiplatelets against collagen than acetylsalicylic acid and, unlike this drug, strongly inhibited thromboxane agonist U46619-induced aggregation. Subacutely administered (5mgkg(-1) day i.p. for 5 days), compounds 6 and 7 protected mice from collagen/epinephrine induced pulmonary thromboembolism at about 20-fold lower doses than acetylsalicylic acid and they prolonged bleeding time like the most part of the other derivatives. The potent antithrombotic activity was coupled with the absence of any lethal and ulcerogenic effect up to 200mgkg(-1) os.

New metal complexes of 4-methyl-7-hydroxycoumarin sodium salt and their pharmacological activity.

Complexes of copper (II), zinc (II), nickel (II), cobalt (II) and iron (III) with 4-methyl-7-hydroxycoumarin sodium salt (Mendiaxon, Hymecromone) were synthesized by mixing of equimolar amounts of the respective metal nitrates and 4-methyl-7-hydroxycoumarin sodium salt in water. The complexes were characterized and identified by elemental analysis, conductivities, IR, 1H NMR spectroscopy and mass spectral data. DTA and TGA have been applied to study the compositions of the compounds. Thermal analysis of the complexes indicate the formation of compounds which correspond to the compositions Met(HL)2 x nH2O, where Met = Cu, Zn, Ni, Co; n = 2, 3 or 4 and Fe(HL)3 x 5H2O. The newly synthesized compounds were assayed for acute intraperitoneal and per oral toxicity, influence on blood clotting time and the most active complex was investigated for spasmolytic activity.

Synthesis of pharmacologically active indoles.

Formylation of 6-methoxy-1-methyl and 5-methyl,2,3-diphenyl-1H-indole (Ib and IX) gave the 5- and 6- carboxaldehyde derivatives (II and X) respectively, which were treated with ethyl cyanoacetate to form the corresponding 2-cyano-3-substituted acrylic acid ethyl ester (III and XI). The latter compounds reacted with hydrazine hydrate, urea and thiourea to form the corresponding 5-amino-4-substituted 2,4,dihydropyrazol-3- one (IV), 6-indolyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carbonitrile s (V and XII) and 6-indolyl-4-oxo-2-thixo-1,2,3,4-tetrahydropyrimidine-5-ca rbonitriles (VI and XIII). Reaction of the 5- and 6-carboxaldehyde derivatives with malononitrile afforded the 2-substituted malononitrile derivatives (VII and XIV). VII and XIV reacted readilly with aromatic ketones to give the 2-amino4,6-disubstituted nicotinonitriles (VIII a,b and XVa,b). The biological activity of compounds Ia, Ib, II, III, IX and X was tested for antiinflammatory, ulcerogenic and antispasmodic activities.

Effect of an adrenergic agonist and a cholinergic antagonist on the airway epithelium.

The ultrastructure of the rabbit tracheal epithelium was studied 30 minutes after intratracheal administration of two puffs of salbutamol and ipratropium bromide, respectively. The injury to the tracheal epithelium due to the treatment with both bronchospasmolytic drugs was considered moderate to severe. In both experimental groups, the degree of goblet cells' stimulation did not differ significantly, the ciliated cells were less damaged compared with the goblet ones and the morphological signs of the impaired self-cleaning ability were revealed.