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Aims: Many patients with glioblastoma (GBM) suffer from comorbid neurological/psychiatric disorders and, therefore, are treated with psychopharmacological agents. Diazepam (DIA) is widely adopted to treat status epilepticus, alleviate anxiety, and inhibit chemotherapy- associated delayed emesis in GBM patients. Even though temozolomide (TMZ) and DIA could be found as possible combination therapy in clinical practice, there are no reports of their combined effects in GBM. Hence, it may be of interest to investigate whether DIA enhances the antitumor efficacy of TMZ in GBM cells. Methods: U87 human GBM was used to examine the effects of combined TMZ and DIA on cell viability, and the oxygen consumption within the cells, in order to evaluate mitochondrial bioenergetic response upon the treatment. Results: The cooperative index showed the presence of antagonism between TMZ and DIA, which was confirmed on long-term observation. Moreover, the level of apoptosis after the TMZ treatment was significantly decreased when administered with DIA (p < 0.001). Concomitant use of TMZ and DIA increased the basal cell respiration rate, the oxidative phosphorylation rate, and maximal capacity of mitochondrial electron transport chain, as well as the activities of complexes I and II, vs. TMZ alone (p < 0.001). Conclusion: Comparing our results with data reported that DIA elicits cell cycle arrest in the G0/G1 phase and favors senescence reveals that DIA diminishes TMZ efficacy in concomitant use in the treatment of GBM. However, due to its great potency to hinder GBM proliferation and metabolism, it could be considered using DIA as maintenance therapy after TMZ cycles.

The effectiveness of the national drug safety monitoring program directly depends on the active participation of healthcare professionals in reporting suspected adverse drug reactions (ADRs). The aim of the study was to explore community pharmacists’ comprehension of pharmacovigilance, their perspectives toward reporting ADRs and investigate the current practice of ADR reporting among pharmacists in Serbia. This descriptive cross-sectional study was performed on a sample of pharmacists in Serbia between November 2019 and March 2020 using a pre-tested questionnaire distributed online. Eligible participants were community pharmacists in Serbia who were willing to participate in the study during the data collection period. Non-parametric statistical tests were performed in the analysis of knowledge, perspectives and ADR reporting. The validity and reliability of the survey were measured by exploratory factor analysis. The median knowledge score was 6 out of 10 (interquartile range 5–7, range 2–10). No significant differences in the knowledge scores of pharmacists were found based on weekly working hours (U = 24,805, p = .374), working experience (χ2 = 4.011, DF = 2, p = .135), being a member of a professional organization (U = 24,312, p = .209), or highest level of pharmacy qualification obtained (χ2 = 3.233, DF = 3, p = .506). Only 28.8% of pharmacists reported ADR at least once a year, while the majority of them have never reported any ADRs. Despite the community pharmacists’ positive attitude toward adverse drug reporting and their role in the process, they show limited knowledge regarding the issue and highly prevalent under-reporting of ADRs. Educational programs are necessary to increase ADRs reporting.

This journal is © The Royal Society of Chemistry. This study was designed to define total protein, phenol and flavonoid content as well as LC-MS/MS phenolic profile related to antioxidant and antidiabetic activity of ethanolic (EtOH) and water extracts of G. pfeifferi and G. resinaceum. G. resinaceum water extract possessed the highest ability to scavenge DPPH and O2-, while the EtOH extract of the same species showed better activity on NO related to other extracts. The highest level of bioactive compounds was determined generally in EtOH extracts. Antidiabetic action was evaluated by the oral glucose tolerance test (OGTT) and histological examination of pancreas and liver in normoglycemic and alloxan-induced diabetic animals. Histological examination of pancreatic tissue demonstrated that G. pfeifferi extracts have protective effects. To conclude, analysed extracts could be considered as a promising candidate for further research with the aim to promote antidiabetic activity, which is for the first time reported for G. pfeifferi.

Graphical abstract, Resveratrol was recognized as the major factor responsible for the beneficial properties of red wine. Several resveratrol-based dietary supplements are available, but their efficacy has not been sufficiently tested. This study was designed to examine the effect of resveratrol supplementation, using a commercially available product, on the metabolic status of experimental animals with induced hyperlipidemia or type 2 diabetes mellitus (T2DM). Hyperlipidemia was induced by feeding the rats a standard pellet diet supplemented with cholesterol. T2DM was induced by adding 10% fructose to drinking water and streptozotocin. Treatment with resveratrol-based supplement improved glycemic control in diabetic animals and significantly decreased serum low-density-lipoprotein (LDL) and triglyceride levels, concurrently increasing the high-density-lipoprotein (HDL) levels in animals with hyperlipidemia. Resveratrol-treated animals had improved tolerance to glucose loading. Supplementation did not induce alterations in parameters of liver and renal function. Findings indicate that commercial resveratrol supplement improves metabolic control in rats with induced hyperlipidemia and T2DM.

This research aimed to examine the effects of pre-treatment with basil extract, prepared as an infusion, on acetaminophen-induced acute liver injury in rats. In vitro analysis of total phenolic, flavonoid content, and antioxidant activity was performed by spectrophotometric methods. Effects of basil extract on oxidative stress parameters, biochemical markers of liver injury, and liver morphology were determined in an in vivo model of acetaminophen-induced liver injury in Wistar rats. Total phenolic content was 52.61 ± 1.35 mg GAE/g of DE, and flavonoid content was 0.5 ± 0.2 mg QE/g of DE. IC50 values ranged from 0.22 to 45.76 μg/ml for DPPH radical, OH radical, H2O2 and lipid peroxidation. Ascorbic acid was used as a positive control. The extract lowered the intensity of lipid peroxidation and potentiated the activity of antioxidant enzymes. In addition, serum levels of aspartate aminotransferase and alanine aminotransferase were significantly lower in basil extract-treated animals in comparison to respective controls.

Microencapsulation represents a process that can create targeted, controlled release kinetics of drugs, thus optimizing therapeutic efficacy. Our group has investigated the impact of this technology on Wistar rats to determine pharmacological efficacy of basil extracts. Animals were treated with water extract of Ocimum basilicum in microvesicles and with combination of basil extracts and 3α,7α-dihydroxy-12-keto-5-cholanate, also known as 12-monoketocholic acid (MKC) acid in microvesicles for 7 days. Alloxan was used to induce hyperglycemia. Pharmacological effects on glycemia were evaluated by measuring blood glucose levels in alloxan-induced diabetic rats. Microvesicles were prepared using the Büchi-based microencapsulating system developed in our lab. The dose of basil extract that was orally administered in rats was 200 mg/kg and the dose of MKC acid was 4 mg/kg as per established protocols. A seven-day treatment with basil aqueous extract, as well as a combination of basil and MKC acid extract in the pharmaceutical formulation, led to a statistically significant reduction in the blood glucose concentration of animals with alloxan-induced hyperglycemia compared to pre-treatment values (p

This study investigated the chemical and nutritional profile and antioxidative properties of cultivated Coprinus comatus. Proximate analysis revealed that C. comatus is rich in carbohydrates, dietary fibres and proteins, and could also be a valuable source of phenolics. Additionally, fat content is low, consisting mainly of polyunsaturated and omega-3 fatty acids. Furthermore, the safety profile of C. comatus is satisfactory, with all elements of toxicological importance within the proposed limits. Oral treatment with C. comatus for 42 days improved the antioxidant capabilities and ameliorated carbon tetrachloride-induced liver damage in rats, marked by decreased serum aminotransferase levels and lipid peroxidation intensity. Glutathione concentrations increased in a dose-dependent manner. Histological morphometric and immunohistochemical analysis confirmed antioxidative and hepatoprotective potential. These findings imply that cultivated C. comatus could be considered a nutraceutical, having beneficial nutrient and therapeutic properties.

The plant world represents an important source of potential therapeutic agents, but concomitant administration of herbal and conventional medications may result in interactions with subsequent beneficial or adverse effects. This study was designed to examine the analgesic effect of thyme tincture and thyme syrup, two commonly used thyme formulations, and their interactions with codeine, paracetamol, pentobarbital and diazepam in mice. The identification and quantification of thymol and carvacrol were carried out by GC/MS and GC/FID. The analgesic activity was studied using a hot plate method. Effects of thyme syrup on diazepam-induced motor coordination impairment in rotarod test and on pentobarbital-induced sleeping time were also determined. Thymol (175.3 µg/mL and 9.73 µg/mL) and carvacrol (10.54 µg/mL and 0.55 µg/mL) concentrations were measured in tincture and syrup, respectively. Thyme syrup and tincture exhibited effective analgesic activity in the hot plate pain model. Pretreatment with thyme formulations reduced analgesic activity of codeine, and potentiated the analgesic activity of paracetamol. Co-administration of thyme formulations has led to potentiation of diazepam and pentobarbital depressive central nervous system effects. Thyme formulations interacted with tested conventional drugs, probably through interference with their metabolic pathways and succeeding altered concentrations and pharmacological effects.

This study was designed to define total protein, phenol and flavonoid content as well as LC-MS/MS phenolic profile related to antioxidant and antidiabetic activity of ethanolic (EtOH) and water extracts of

Pycnogenol® (PYC) has already being used as a food supplement and herbal medicine due to its potent antioxidant properties. The aim of the present study was to examine the protective effect of PYC on acetaminophen-induced acute liver injury in rats. The effect of PYC on acetaminophen-induced hepatotoxicity in rats was examined by determining biochemical parameters, in vitro antioxidant activity, histological assessment, and oxidative status in liver homogenates. The best antioxidant properties were demonstrated in methanolic extracts. Seven-day pretreatment with PYC suppressed elevation of CYP2E1 protein expression induced by administration of toxic dose of acetaminophen. PYC at 50 mg/kg showed the ability to significantly decrease malondialdehyde (MDA) level compared with the group received acetaminophen. Xanthine oxidase (XOD) enzyme activity was significantly elevated in acetaminophen-treated group compared with control, whereas concomitant administration of PYC in a dose of 50 mg/kg significantly reduced activity of this enzyme. Significant decrease of glutathione (GSH) hepatic content in acetaminophen-intoxicated rats compared with the control rats was improved by concomitant administration of PYC at 50 mg/kg. Protective effect of PYC on acetaminophen-induced acute liver injury in rats has showed the best in vitro antioxidant potential expressed in methanolic extract and consequent histological assessment and oxidative status in liver homogenates.

Background. Attitudes towards conventional and complementary medicine among future healthcare professionals can impact their future pharmacotherapy practice. This study aimed to determine the prevalence and predisposing factors related to self-medication among medical and pharmacy students. Methods. This cross-sectional questionnaire-based study was performed at the Faculty of Medicine, University of Novi Sad, Serbia, on first- and final-year students of medicine and pharmacy. The multivariate Poisson regression model with robust variance was used to identify the main predictors of self-medication. Results. The overall self-medication prevalence in the past year was 81.3%. Independent risk factors for self-medication identified in the regression analysis were the final study year, housing condition, i.e., living in a leased apartment or in a student dormitory in comparison to living with parents, and cigarette consumption. The conventional drugs were the most frequently used, mostly for the symptoms of cold and pain. Final-year students had more confidence in conventional medicines than in herbal drugs and were more aware of the risks of their concomitant use. Conclusion. Self-medication is highly prevalent among students of medical sciences, especially among final-year students. Increased medical knowledge led to the higher awareness of the drug interaction risks.

Glioblastoma (GBM) is one of the deadliest primary brain neoplasm, heavily infiltrated with tumor-associated microglia/macrophages (TAM), which has received a great deal of interest. Bearing in mind that the number of peripheral macrophages by the 14th day is negligible, in our study TAM were referred to as microglia. Here we evaluated histopathological characterization of TAM and kinetics of their infiltration in U87 orthotopic GBM, a commonly used model in preclinical research. To mimic different stages of GBM growth, we evaluated three-time points. Our data showed that the highest areal density of TAM was 7 days after GBM inoculation, with ability to proliferate early after initiation of GBM growth. The areal density of TAM within the tumor correlated with GBM growth and proliferation. Moreover, microglia underwent substantial morphological changes upon exposure to GBM cells. A transition from ramified morphology in peritumoral area to ameboid shape with larger soma and shortened, thick branches in the tumor core was observed. Higher areal fraction of blood vessels also correlated with the areal density of TAM. Given these pro-invasive features of microglia, this GBM model represents a good basis for further testing microglia as a target and new strategy to fight with.

Aniseed (Pimpinella anisum L., Apiaceae) and its essential oil (EO) have been widely used. Because there are some data about the impact of aniseed EO on drug effects, this survey aimed to assess the potential of pharmacokinetic herb-drug interaction between aniseed EO and acetaminophen and caffeine in mice. The chemical analysis (gas chromatography-mass spectrometry) of aniseed EO has confirmed trans-anethole (87.96%) as the main component. The pharmacokinetic studies of intraperitoneally (i.p.) and orally applied acetaminophen (200 mg/kg) and caffeine (20 mg/kg) were performed in mice after 5 days of oral treatment with human equivalent dose of aniseed EO (0.3 mg/kg/day). The analysis of pharmacokinetic data showed that in the group treated by aniseed EO, the significant decrease in the peak plasma concentration of acetaminophen after oral application (p = 0.024) was revealed when compared with control group and the reduction of systemic exposure to the drug after oral application (74 ± 32% vs. 85 ± 35% in the control) was noted. The bioavailability of orally applied caffeine was also significantly decreased (p = 0.022) after the EO treatment in comparison with the control (57 ± 24% vs. 101 ± 29%). Therefore, the compromised therapeutic efficacy of acetaminophen and caffeine during the usage of aniseed EO preparations should be considered.

This study investigated the chemical and nutritional profile and antioxidative properties of cultivated Coprinus comatus . Proximate analysis revealed that C. comatus is rich in carbohydrates, dietary fibres and proteins, and could also be a valuable source of phenolics. Additionally, fat content is low, consisting mainly of polyunsaturated and omega-3 fatty acids. Furthermore, the safety profile of C. comatus is satisfactory, with all elements of toxicological importance within the proposed limits. Oral treatment with C. comatus for 42 days improved the antioxidant capabilities and ameliorated carbon tetrachloride-induced liver damage in rats, marked by decreased serum aminotransferase levels and lipid peroxidation intensity. Glutathione concentrations increased in a dose-dependent manner. Histological morphometric and immunohistochemical analysis confirmed antioxidative and hepatoprotective potential. These findings imply that cultivated C. comatus could be considered a nutraceutical, having beneficial nutrient and therapeutic properties.

Introduction: Rosemary essential oil can be used for treating dyspepsia, mild spasmodic disorders of thegastrointestinal tract, externally as analgetic in muscular and articular pain and minor peripheral circulatory disorders.It is important to explore its analgesic potential and its influence on the pharmacodynamic and pharmacokineticproperties of paracetamol. Methodology: Rosemary essential oil was applied to mice orally (doses: 10 and 20 mg/kgb.w.) for pharmacodynamic tests for seven days. Rats treated with rosemary essential oil for seven days orally (5 and10 mg/kg b.w.) were used for the examination of the influence of essential oil on the pharmacokinetic properties ofparacetamol (applied on the 7th day p.o. or i.v.). Pharmacokinetic parameters were analyzed in blood samples obtainedfrom rats' tail veins, with the HPLC method. Results: The essential oil of rosemary shows analgetic properties. Therosemary essential oil increases pharmacological effects of paracetamol and does not change paracetamolbioavailability. Conclusion: The herbal drugs could change the pharmacodynamic and the pharmacokinetic propertiesof classical drugs and they could also change the safety of classical drugs use in human population.

The aim of this study was to evaluate zinc gluconate as a treatment option in patients with symptomatic migratory glossitis (MG). Using simple random sampling, 28 non-psoriatic patients with symptomatic MG were divided into a test and control group. The test group took 20 mg/day of chelated zinc gluconate for one month, and was put on a diet rich in zinc. The control group was only put on a diet rich in zinc. Changes in the size of red atrophied areas (width and length) and the intensity of symptoms were evaluated as primary and secondary outcomes, respectively, at baseline, after therapy, and one month later. In the test group, the mean value of the red atrophy area width and length displayed some significant reduction as a primary outcome. There were no significant changes in the size of red patches in the control group. Secondary outcome showed that the intensity of subjective symptoms in the test group significantly decreased (P=0.042) compared with controls. The filiform papillae had partially or completely regenerated in 85.7% of cases in the test group and in 23.1% of the controls (P=0.001). Red patches with raised keratotic rims may have healed spontaneously and reappeared in constantly changing patterns that are typical for MG. This phenomenon was not observed in patients supplemented with zinc, and new atrophy areas occurred in only one case. Low-dose zinc gluconate.

Background/Aim. The overuse of antibiotics unnecessarily exposes patients to risk of side effects, encourages reconsultation for similar problems and enhances antimicrobial resistance. The use of antibiotics in the year 2011 in Montenegro was high (39.05 Defined Daily Dose ? DDD/1,000 inhabitants/day), but it was not considered in relation to the frequency of bacterial diseases. The aim of our study was to determine the degree of conformance between the amount of outpatient antibiotic consumption and the reported prevalence of outpatient bacterial infections in the Republic of Montenegro. Methods. Data on the use of antibacterial drugs was obtained from the Agency for Medicines and Medical Devices of Montenegro for the year 2012. The amount of antibiotics was calculated using the Anatomic Therapeutic Chemical (ATC) DDD methodology. Data on the prevalence of outpatient infective disease was obtained from the Health Statistical Yearbook 2012 of Montenegro and it was expressed per 1,000 inhabitants. Results. A total of 30.34 DDD/1,000 inhabitants/day of antibiotics in outpatients were prescribed in Montenegro in 2012, with penicillins being most frequently prescribed. Amoxicillin and amoxicillin with clavulanic acid were the most frequently used antibiotics. The prevalence of outpatient bacterial infections was 6,745 cases or 10.87/1,000. The most frequent infections were respiratory tract infections. Less than 50% of the prescribed amount of antibiotics were prescribed in accordance with national guidelines on treatment of bacterial infections. Conclusion. Use of antibiotics in Montenegro in 2012 was more than double than necessary according to prevalence of bacterial infections and average duration of treatment. The structure of antibiotics was not in full compliance with the national good practice guidelines, but it was in accordance with data on bacterial antibiotic resistance in outpatient practice. It is necessary to initiate measures to rationalize the use of antibiotics both in terms of quantity and in terms of the structure of the most used antibiotics.

The aim of this study was to evaluate zinc gluconate as a treatment option in patients with symptomatic migratory glossitis (MG). Using simple random sampling, 28 non-psoriatic patients with symptomatic MG were divided into a test and control group. The test group took 20 mg/day of chelated zinc gluconate for one month, and was put on a diet rich in zinc. The control group was only put on a diet rich in zinc. Changes in the size of red atrophied areas (width and length) and the intensity of symptoms were evaluated as primary and secondary outcomes, respectively, at baseline, after therapy, and one month later. In the test group, the mean value of the red atrophy area width and length displayed some significant reduction as a primary outcome. There were no significant changes in the size of red patches in the control group. Secondary outcome showed that the intensity of subjective symptoms in the test group significantly decreased (P=0.042) compared with controls. The filiform papillae had partially or completely regenerated in 85.7% of cases in the test group and in 23.1% of the controls (P=0.001). Red patches with raised keratotic rims may have healed spontaneously and reappeared in constantly changing patterns that are typical for MG. This phenomenon was not observed in patients supplemented with zinc, and new atrophy areas occurred in only one case. Low-dose zinc gluconate supplementation may have a positive therapeutic effect on tongue epithelium regeneration and symptomatology in patients with MG. in our region were consistent with those from other studies.

Dietary supplements based on soybean extracts are widely used for a myriad of indications, but there is evidence that isoflavones, components of these supplements affect cytochrome activity. Alteration in pharmacodynamics of cytochrome-substrate drugs after co-administration with dietary supplements based on soybean extracts offers information on clinically significant herb-drug interactions that can cause unanticipated adverse reactions or therapeutic failures. Moreover, herbal drugs can contribute to the development and severity of drug-induced liver injury. Therefore, the aim of this study was to examine the effect of commercially available dietary supplements based on soybean extracts on liver and renal function, oxidative stress and pharmacodynamics of several conventional drugs. After quantification of isoflavones in the supplement using HPLC-DAD, pharmacological tests such as hot-plate, rotarod, pentobarbital-induced sleeping time were performed on Swiss albino mice treated with soybean extracts and several central nervous system acting drugs. Liver, renal function, and oxidative status in liver homogenates were determined in healthy and Wistar rats subjected to oxidative stress with CCl4. Dietary supplements based on soybean extracts weakened the analgesic activity of codeine, significantly potentiated diazepam-induced motor coordination impairment at 10th and 30th minute after diazepam administration, but had the opposite effect on alprazolam effect. Soybean extracts pretreated group also exhibited significantly shorter pentobarbital sleep induction and sleeping time. Soybean extracts administration did not affect the liver and renal function and ameliorated oxidative stress caused by CCl4. Despite exhibiting no negative effects on liver and renal function and demonstrated antioxidant in vivo potential, the safety of soybean extracts in addition to conventional drugs is questionable. The results of our study implicate the potential of dietary supplements based on soybean extracts for serious herb-drug interactions.

Background Worldwide data indicate that self-medication is frequently used inappropriately. Although self-medication is encouraged in most of the countries by introducing over-the-counter drugs, it bears the risk of misuse of drugs issued on prescription due to low observance of legislation of medicines requiring prescription by some pharmacies. Objective The objective of this study was to explore the self-medication practice, with an emphasis on self-medication with prescription-only medications. Setting Households in Novi Sad city, Serbia. Method The study was conducted over 8 month period (December 2011–July 2012) and involved a random sample of households. The questionnaire-based study and personal insight into household drug supplies was performed by a trained interviewer. Main outcome measure Number of drugs obtained without prescription or without consulting a physician in surveyed households. Results The total number of drug items present in the 383 households was 4,384 with a median of 11 drugs per household. More than a half of drugs in households were prescription-only medication (58.5 %). Approximately one third of prescription-only medications were obtained without prescription. The most common drugs obtained without prescription were anti-inflammatory and antirheumatic products and antibacterials for systemic use. Ibuprofen and diclofenac were the most common self-medicated drugs. Number of prescription-only medications bought with ought prescription was significantly higher in households with children under 12 years of age compared to other types of households. Conclusion Our survey indicated that self-medication with prescription drugs appeared to be a rather common practice, which is far away from the concept of “responsible self-medication”, especially regarding antibiotics.

Humans and guinea pigs are unable to produce vitamin C, with deficiency resulting in a well-known disorder of collagen synthesis. Pial basement membrane structure preservation is essential in the proper migration of neurons. In our study, intrauterine deprivation of vitamin C in guinea pig fetuses led to a collagen synthesis disorder, weakness, and finally a breach of pial basement membrane. We found excessive migration of the external germinal layer cells into the subarachnoid space of the cerebellum through defects in the pial basement membrane. The changes ranged from focal rupture of pial basement membranes to their complete disintegration. The loss of proper folia formation resulted in macroscopically visible flattening of the cerebellar surface. Different grades of dysplastic changes in the folia of the cerebellar cortex were observed in 2 experimental groups assigned different limits to mark the time of commencement and duration of vitamin C deprivation. The most severe form of dysplastic changes was characterized by marked irregularity of the cerebellar cortex similar to that in lissencephaly type II. Thus, prenatal vitamin C deficiency represents a novel animal model to study the effects of collagen synthesis on development of breaches in the pial basement membrane, disordered migration of neurons, dysplasia of cerebellar cortex, and the pathogenesis of lissencephaly.

Aim: The aim of this study is to investigate the influence of bile salts, sodium cholate, sodium 12-ketocholate and sodium dehydrocholate, as excipients in ranitidine, aminophylline and phenobarbital tablets on dissolution rate. Methods: Four groups of tablets (control without bile salts and three investigational groups containing different bile salts) were prepared for three different drug substances: ranitidine, aminophylline and phenobarbital. Dissolution rate was measured. Results: Dissolution rate is increased significantly in all investigational groups comparing to the control group in all three drug substances. Discussion: Presented results are very favourable and encouraging in case of dissolution enhancing and should be further investigated, especially in drug substances that are classified in class II and IV as per Biopharmaceutical Classification System (BCS) classification. Conclusion: Bile acid salts are very promising excipients, proven to act as surfactants and as lubricants. Running title: Bile salts as excipients in ranitidine, aminophylline and phenobarbital tablets.

Herb-drug interactions are an important safety concern and this study was conducted regarding the interaction between the natural top-selling antidepressant remedy Hypericum perforatum (Hypericaceae) and conventional drugs. This study examined the influence of acute pretreatment with different extracts of Hypericum perforatum from Serbia on pentobarbital-induced sleeping time, impairment of motor coordination caused by diazepam and paracetamol pharmacokinetics in mice. Ethanolic extract, aqueous extract, infusion, tablet and capsule of Hypericum perforatum were used in this experiment. The profile of Hypericum perforatum extracts as well as paracetamol plasma concentration was determined using RP-HPLC analysis. By quantitative HPLC analysis of active principles, it has been proven that Hypericum perforatum ethanolic extract has the largest content of naphtodianthrones: hypericin (57.77 µg/mL) and pseudohypericin (155.38 µg/mL). Pretreatment with ethanolic extract of Hypericum perforatum potentiated the hypnotic effect of pentobarbital and impairment of motor coordination caused by diazepam to the greatest extent and also increased paracetamol plasma concentration in comparison to the control group. These results were in correlation with naphtodianthrone concentrations. The obtained results have shown a considerable influence of Hypericum perforatum on pentobarbital and diazepam pharmacodynamics and paracetamol pharmacokinetics.

Background/Aim. It is known that bile acids improve the absorption, bioavailability and pharmacodynamic characteristics of some drugs. Morphine analgesia is produced by activation of opioid receptors within the central nervous system (CNS) at both spinal and supraspinal levels. Since a morphine molecule contains 3 polar groups and therefore hard to transfer through the blood-brain barrier, the aim of the study was to examine the potential influence of bile acids derivates, namely sodium salt of monoketocholic acid (MKH-Na) and methyl ester of monoketocholic acid (MKH-Me), on analgesic effect of morphine. Methods. White male mice of NMRI-Haan strain, with body weight of 20-24 g, were used in this study. The analgesic effect of morphine (administered by subcutaneous and intramuscular route in a dose of 2 mg/kg), with and without pretreatment with MKH-Na (4 mg/kg) and MKH-Me (4 mg/kg) was estimated by the hot plate method. Results. Administration of MKH-Me prior to subcutaneous administration of morphine increased the morphine analgesic effect but the increase was not statistically significant. At the same time administration of MKH-Na did not affect morphine analgesic effect. The analgesic effect of morphine increased when administered intramuscularly 20 min after MKH-Me administration. When compared with the group of animals treated only with morphine, a statistically significant increase in analgesic effect was detected 10, 30, 40 and 50 min after morphine administration (p < 0.05). Pretreatment with MKH-Na did not affect morphine analgesic effect. Conclusion. According to the results of this study it can be presumed that after intramuscular morphine administration methyl ester of monoketocholic acid increases morphine transport into the central nervous system and consequently the analgesic effect, as well. Further research on bile acids-morphine interaction both in vitro and in vivo is necessary to completely elucidate the mechanism of this interaction and increase in the morphine analgesic effect. [Projekat Ministarstva nauke Republike Srbije, br. 41012: Interactions of xenobiotics and the impact on biomedical system i br. 172050: Development and application of advanced chromatographic and spectroscopic techniques in analysing xenobiotics and the mechanisms of their decomposition in biotic and abiotic samples]

Diabetes mellitus is a chronic disease characterized by abnormal carbohydrate, lipid and protein metabolism due to a lack of insulin or reduced target cell sensitivity to insulin. Stevia rebaudiana is an important source of biochemically active substances with proven anti-diabetic effect. The aim of this study was to determine anti-diabetic effects of the low dose of stevioside in NMRI Haan mice. Aqueous stevioside solution (20mg/kg body weight) was administered by oral route of administration. Anti-diabetic effect of stevioside was estimated by oral glucose tolerance test, adrenaline test after a 10day stevioside treatment, and alloxan induced hyperglycaemia in mice (two experimental groups, 10day stevioside treatment before and after alloxan administration). Aqueous stevioside solution prevented significant increase in glycaemia in oral glucose tolerance test (9.22±1.13 to 9.85±1.32mmol/l, P

Silymarin is a complex of five major compounds, and silibinin is the most biologically active component of the complex. The aim of this study was to investigate, evaluate and confirm the potential cardioprotective and hepatoprotective effects of administration of silymarin, rich in silibinin, at a dose of 60 mg/kg orally for a time-span of 12 days on doxorubicin induced toxicity in male Wistar rats. The in vivo model was used to explore whether silymarin could prevent damage of liver and heart tissue induced by doxorubicin administered every other day at dose of 1.66 mg/kg intraperitoneally for twelve days. In the study the change of body weight, ECG changes, biochemical parameters of oxidative stress, serum activity of alanine and aspartate transaminase, lactate dehydrogenase, creatine kinase and histological preparations of heart and liver samples of treated animals were examined. According to physiological, pharmacological, microscopic and biochemical results, we confirmed that at the examined dose, silymarin exhibits a protective influence on the heart and liver tissue against toxicity induced by doxorubicin.

The pharmacodynamic effect of a 7-day oral treatment with a suspension of Coprinus comatus at doses of 0.835 and 1.670 g/kg in rats was studied. Changes in body weight, bile secretion and hypoglycaemic action were examined together with antipyretic activity and paw oedema tests. Such treatments resulted in a significantly lower increase in the body weight of tested animals (15.73 ± 8.36 g/rat in the untreated group, 8.44 ± 8.23 g/rat (p < 0.05) and 3.18 ± 7.93 g/rat (p < 0.05), for C. comatus 0.835 and 1.67 g/kg, respectively). Hypoglycaemic action was evident only in the glucose load test (6.79 ± 0.61 to 9.70 ± 1.16 (p < 0.05) in the untreated group and 6.47 ± 0.35 to 7.27 ± 0.76 for C. comatus 1.67 g/kg). Histological examination of pancreas cross-sections suggested certain protective functions of the mushroom suspension in alloxan poisoning. In the antipyretic test, a significantly lower increase in body temperature was observed in the mushroom-pretreated rats. In the paw oedema test, no decrease in oedema induced by formalin injection was observed following treatment with C. comatus.

The discovery of agents which induce regeneration of pancreatic beta cells and insulin secretory potential would be useful to develop new therapeutic approaches to treat diabetes. We investigated effects of mushroom Coprinus comatus on regeneration Langerhans islets after alloxan treatment. In this study, on immunohistochemically stained histological preparations (insulin and glucagon) was apply morphometric analysis of of rat pancreatic tissue to examined: a) the degree of regenerative characteristics of islets of Langerhans in alloxan induced diabetes, b) the impact of mushroom Coprinus comatus on pancreatic tissue and c) the impact of the mushroom Coprinus comatus regenerator properties Langerhans islets after administration of alloxan. Suspension of mushroom Coprinus comatus was applied in three time periods (7, 21, and 42 days) at three different doses of 0.835, 1.67, and 3.34 g/kg. Morphometric parameters were used: the volume density of endocrine and exocrine part of islets per pancreatic tissue; the mean corrected islet diameter , the numerical density of alpha and beta cell per islet profile, the number of alpha and beta cell type per islet, the corrected mean nuclear diameter, and corrected mean nuclear volume. The results showed that the application of alloxan confirmed so far above the selective destruction of beta cells with a relative degree of regeneration that occurred in animals sacrificed after 4, 5, 7, 21 and 42 days after application of alloxan. In the experimental group of animals that received only the mushroom Coprinus comatus, the application dose of 1.67 g/kg is observed to increase in alpha and beta cells, while the other groups showed no significant difference compared with the control group. In rats which after inducing diabetes with alloxan and then applied preparation mushroom Coprinus comatus can be observed much greater regeneration of islets compared to pure alloxan induced group. Morphometric analysis revealed a proliferation of alpha cells and a significant increase in beta cell mass. In addition to clearly immunopositive cells were observed and transitional cell forms between alpha and beta cells, indicating the occurrence of the process of transdifferentiation of one cell into another form. Therefore, regeneration of islets covered the two processes, one is the proliferation of alpha cell population and the second is the appearance of new beta cells for which assume that the resulting direct conversion of alpha cells. Stimulating effect of mushroom Coprinus comatus on the regeneration of the islets of Langerhans gives us precious, so far no published data on the impact of these mushrooms in vivo in living systems.

Aniseed (Pimpinella anisum L., Apiaceae) and its essential oil (EO) have been widely used. Because there are some data about the impact of aniseed EO on drug effects, this survey aimed to assess the potential of pharmacokinetic herb-drug interaction between aniseed EO and acetaminophen and caffeine in mice. The chemical analysis (gas chromatography-mass spectrometry) of aniseed EO has confirmed trans-anethole (87.96%) as the main component. The pharmacokinetic studies of intraperitoneally (i.p.) and orally applied acetaminophen (200 mg/kg) and caffeine (20 mg/kg) were performed in mice after 5 days of oral treatment with human equivalent dose of aniseed EO (0.3 mg/kg/day). The analysis of pharmacokinetic data showed that in the group treated by aniseed EO, the significant decrease in the peak plasma concentration of acetaminophen after oral application (p = 0.024) was revealed when compared with control group and the reduction of systemic exposure to the drug after oral application (74 ± 32% vs. 85 ± 35% in the control) was noted. The bioavailability of orally applied caffeine was also significantly decreased (p = 0.022) after the EO treatment in comparison with the control (57 ± 24% vs. 101 ± 29%). Therefore, the compromised therapeutic efficacy of acetaminophen and caffeine during the usage of aniseed EO preparations should be considered.