[ANTIOXIDANT ACTIVITY OF XYMEDONE IN RATS WITH CHRONIC AUTOIMMUNE INFLAMMATION].

Effects of drug xymedone (in comparison to ionol) in a group of 32 white rats with experimental model of chronic autoimmune inflammation of rat paws (induced by Freund's adjuvant) were studied by measuring the volume of paw edema and determining the levels of lipid peroxidation (LPO) products and the activity of antioxidant enzymes in various tissues. Chronic autoimmune inflammation induced by Freund's adjuvant was characterized by the LPO intensification and disturbances of the level of antioxidant enzymes. Intragastric administration of xymedone (2,2-dihydro-4,6-dimethyl,-N-(ß-oxy-ethyl)-2-pyrimidon) at a dose of 169 mg/kg and reference drug ionol (2,6-ditretbutyl-4-methylphenol) at a dose of 220 mg/kg increased the activity of serum antioxidant enzymes by 19% and 11%, respectively, decreased the serum level of nitrite ion by 62% and 50%, and reduced the levels of LPO products in rat blood and homogenates of liver, kidney, and spleen by up to 80% (p < 0.05).

[NEW APPROACH TO COMPARING NSAID EFFICACY BY LONG-TERM MONITORING OF EXPERIMENTAL INFLAMMATION MODELS.]

Experiments on inflammatory edema modeling by sub-plantar injection of carrageenan lambda (1 %) and formalin (2 %) showed substantial differences betwe- en the two models during long-term observation, including irreversible damage caused by formalin (at reversible carrageenan action) and high intensity of for- malin edema (in contrast to carrageenan edema) in mice. We propose a new approach to evaluation of the so-called total inflammatory burden (experimental analog of disease outcome) by calculating the area under the inflammation intensity versus time curve. With the use of this approach, we showed the absence of any effect of conventional NSAIDs (naproxen, diclofenac. indomethacin) on the total inflammatory burden induced by carrageenan or formalin injections in mice and rat paw edema models. These results show the need for using new approaches in the search for potential anti-inflammatory agents.

Towards the rational use of medicines.

BACKGROUND: Rational use of medicines remains to be one of the most challenging problems in health systems worldwide [1, 2]. ABC/VEN-analysis has been recommended for use by the World Health Organization (WHO) and has been used in health care practice globally since 1981. It represents the simple and effective method of analysis of medicine expenditures, identifying priority groups of medicines, the use of which, when improved, may provide the greatest clinical and economic impact. ABC analysis provides an accurate and objective picture of budget expenditures on medicines. VEN-analysis helps to prioritize between various medicines in their selection for procurement and use within a drug supply system [3-5]. OBJECTIVE: To assess the impact of introduction of evidence-based principles in the practice of medicine procurement and use on budget expenditures on medicines of a multidisciplinary health facility for the period of four years (2011-2014). METHODS: ABC/VEN analysis was carried out in a multidisciplinary health facility with over 1000 beds (an average number of beds for three years), which is responsible for provision of care to the population of about 1.4 million people. The analysis was carried out on the basis of information on medicine expenditures for 4 years: 2011 (1st year), 2012 (2nd year), 2013 (3rd year) and 2014 (4th year). When assigning VEN categories of medicines we used expert method: assignment of categories was carried out by clinical pharmacologists after reviewing all available evidence on effectiveness, safety and cost-effectiveness compared to other drugs in this group. In 2013, we implemented educational intervention, including detailed discussion of the results of the ABC/VEN-analysis for the years 2011-2012 from the standpoint of evidence-based pharmacology and recommendations for medicine procurement. In 2014, we delivered training workshop for the heads of clinical departments on evidence-based principles in clinical pharmacology and rational use of medicines. RESULTS: Medicines expenditures of the studied health facility for the year 2014 were less than for the year 2013, which was the important decrease reversing the trend of increasing medicines expenditures of the last three years: 2011 - 59,868,963 roubles; 2012 - 85,324,084 roubles, 2013 - 107 303 390 roubles, and 2014 - 74,416,692 roubles. The number of International Non-proprietary Names (INN) of medicines used in 2014 was 519, which was the highest number for the four years of the study: 2011 - 429 INN, 2012 - 432 INN, 2013 - 513 INN, and 2014 - 519 INN. Nearly 40% of the funds spent in 2014 on medicines have been used for Vital medicines: 2011 - 26%, 2012 - 39%, 2013 - 25%. Expenditures on Non-essential medicines in 2014 were about the same as in previous years - 14% of total medicine expenditures: 2011 - 16%, 2012 - 13%, 2013 - 15%. However in absolute numbers (roubles) expenditures on non-essential medicines decreased compared to the years 2013 and 2012: 2011 - 9,428,135 roubles, 2012 - 11,129,388 roubles, 2013 - 15,578,325 roubles, 2014 - 10,616,023 roubles.Expenditures on solutions for infusion (sodium chloride, Ringer's solution, dextran, glucose, hydroxyethyl starch) decreased as compared to the year 2013, but still remained high, thus indicating on the abuse of parenteral methods of drug administration. The portion of expenditures on isotonic sodium chloride solution and hydroxyethyl starch in 2014 decreased compared to the year 2013. We found a positive trend in the structure of expenditures on antibacterial agents: in 2014 expenditures on fluoroquinolones decreased nearly fivefold compared to 2013, expenditures on cephalosporins also decreased, but not so dramatically. However, there was a significant increase in expenditures on carbapenems, more than twofold compared with the year 2013. In 2014 we noted a twofold decrease in expenditures on medicines affecting blood, including antithrombotic agents, hemostatics and antianemic medicines, as compared to the values of the year 2013. In 2014 there was also a decrease in expenditures of cardio-vascular medicines, medicines affecting nervous system, alimentary tract and metabolism. CONCLUSIONS: Introduction of evidence-based principles through educational interventions at a multidisciplinary health facility resulted in a number of changes towards more rational medicine use. Regular educational interventions for practicing physicians and heads of clinical departments of health facilities that promote rational prescribing are needed.

Anti-inflammatory and anti-oxidant properties: Is there a link?

BACKGROUND: It is believed that the anti-inflammatory activity of medicines is closely related to their antioxidant activity. However, in clinical practice rigorous evidence-based medicine approach fails to reveal important effects of antioxidants on patient important outcomes in inflammatory disorders, as has been shown by a number of Cochrane reviews [1-3]. OBJECTIVE: To evaluate anti-inflammatory and antioxidant effects of newly developed pharmacological agents: dimephosphone and its structural analogues ephorane and mephoprane, and xymedon, in comparison with prednisolone and etidronate in experimental animal model of adjuvant arthritis. METHODS: Experiments were conducted in 64 white mongrel rats of both sexes weighing 180-200 g, which were divided into 8 groups 8 rats each (4 males and 4 females each), kept under standard vivarium conditions with certified feeding ration (kombikorm). The study was approved by the local ethics committee. We induced adjuvant arthritis by administration under the plantar aponeurosis of the left hind paw of 0.1 ml of Freund's adjuvant (Sigma) in rats of 7 study groups. The groups were as follows: 1st group - intact animals (control); 2nd group - animals to whom the solvent (distilled water) was administered with intra-gastric tube in corresponding volume (control of the model); 3rd - 8th study groups, in which animals were administered with study agents each at a dose of 1 mmol/kg body weight: dimephosphone, ephorane, mephoprane, xymedon, etindronate and prednisolone. The intensity of the modeled arthritis was determined by measurements of paw volumes with plethysmometer (UgoBasile). We calculated the difference in rat paw volume before the administration (baseline) and after administration of Freund's adjuvant at 3, 7, 11, 15, 20, 27, 31, 38, 41 days. The development of secondary arthritis was documented by the increase in volume of both hind and fore paws and tails. On the 41st day of the experiment the animals were sacrificed under light ether anesthesia and exsanguinated. The blood was used to determine the activity of catalase and peroxidase, the content of the total, reduced and oxidized glutathione, the level of ceruloplasmin, conjugated dienes of unsaturated fatty acids (DC), TBA-interacting products (MDA), and the total antioxidant activity of serum (AOA). The results were processed statistically using the Student's t-test. RESULTS: The primary reaction to the Freund's adjuvant in a form of swelling of the ankle joint of the left hind paw was observed at 24 hours after its injection. External clinical manifestations of the modeled disease were more pronounced on the third day: local inflammatory reaction (redness, swelling, ulceration) was seen in all the animals at the injection site with the increase of the paw volume. On the 11th day of the experiment 20% of the animals developed secondary arthritis. The study agents dimephosphone, ephorane, and prednisone exerted anti-inflammatory effect decreasing the volume of left hind paws by 45%, 46% и 27% respectively on the 40th day of experiment. Mephoprane did not affect the primary inflammatory response to the Freund's adjuvant (rats' left hind paws), however it reduced the volume of the contralateral right paw (secondary arthritis) by 90% on the 20th day of the experiement. This ant-inflammatory effect was accompanied by documented antioxidant activity in case of dimephosphone, ephorane, prednisolone, but not mephoprane. Dimephosphone reduced the levels of lipid peroxidation products in rats blood by 46% (DC) and by 25% (MDA). Ephorane also reduced the levels of lipid peroxidation products in the blood by 46% (DC) and by 25% (MDA), increasing the level of glutathione by nearly half, both the total and the reduced form. Prednisolone reduced the level of lipid peroxidation products in blood by 61% (DC), but not the TBA-interacting products. Mefopran did not affect the blood level of lipid peroxidation products. Xymedon and etidronate showed no anti-inflammatory effect. Xymedon demonstrated anti-oxidant properties, decreasing the blood levels of lipid peroxidation products, while etidronate seemed to behave in pro-oxidant mode, increasing the blood levels of lipid peroxidation products. CONCLUSIONS: The effects of studied agents on the intensity of inflammation and lipid peroxidation were inconsistent. The results of the study did not show a clear link between anti-inflammatory and anti-oxidant activity. Further research in potential anti-inflammatory activity of new drugs exhibiting antioxidant properties needs to be done before recommending their use in clinical practice.

Rat paw oedema modeling and NSAIDs: Timing of effects.

BACKGROUND: Non-steroidal anti-inflammatory agents (NSAIDs), steroids and representatives of other pharmacological groups [1, 2] are widely used for pharmacological regulation of inflammation. However, their anti-inflammatory effects are accompanied by serious adverse reactions [3, 4]. There was a hope that newer NSAIDs, selective inhibitors of COX-2, would be safer, but their longer-term use appeared to cause an increased risk of heart attacks and stroke [5]. Carrageenan rat paw oedema model is traditionally used for search and development of new NSAIDs with assessment of effects after 3 to 5 hours after oedema induction [6, 7], neglecting longer-term effects [8]. OBJECTIVE: To compare effects of traditional NSAIDs (indomethacin, naproxen) on the development, duration and intensity of carrageenan rat paw oedema. METHODS: Carrageenan paw oedema was induced in 18 rats by sub-plantar injection into the right hind paw of the animals of 0.1 ml of 1% aqueous gel of carrageenan-λ (22049 SIGMA λ-Carrageenan plant-mucopolysaccharide, Sigma-Aldrich). We assessed the intensity of the oedema development and its duration by measurements of rat paw volume using plethysmometer 37140 (UgoBasile, Italy). Measurements were made prior to induction of oedema (base-line volume) and at 1, 2, 3, 4, 5, 24, 48, 72, 96, 120, 144, 168 and 192 hours after sub-plantar carrageenan injection. Calculating the percentage of increase in paw volume assessed the intensity of the oedema. The base-line paw volume was taken for 100%.Animals were divided into 3 groups of 6 rats each; group 1: control (solvent); group 2: naproxen 15 mg/kg and group 3: indomethacin 10 mg/kg. These doses are known as ED50 (effective doses 50) on carrageenan rat paw oedema with single-dose NSAIDs administration [9]. To get the most accurate estimate of the intensity of the simulated by carrageenan inflammatory response and the potential effects of some NSAIDs with their longer-term use we calculated areas under the curve «increase in paw volume - time¼ using standard method of numerical integration - trapezoidal method. Statistical analysis was performed using Microsoft Office Excel 2007 with the calculation of arithmetic means M, their standard deviations (δ) and standard errors (m). We applied Student's t-test and accepted as significant the differences with P values equal to or less than 0.05. RESULTS: The inflammatory reaction induced by carrageenan, developed in a form of swelling/oedema with an increase in the rat paw volume up to 55% of the baseline volume. The maximum volume of oedema was observed in the control group at 3 h after the injection of carrageenan, which is in accordance with the literature data on the development of carrageenan paw edema in rats [10, 11]. Naproxen at a dose of 15 mg/kg showed anti-inflammatory activity at 1, 2, 3, 4 and 5 hours after administration of carrageenan with suppression of oedema development by 59, 81, 73, 60 and 39% (p = 0.03; 0.001; 0.001; 0.001 and 0.01), respectively. There was no oedema inhibition by naproxen at later time-points. Indomethacin at a dose of 10 mg/kg showed anti-inflammatory effect at 2, 3, 4, and 5 hours after carrageenan oedema induction with inhibition of oedema development by 54, 54, 54 and 33% (p = 0.01, 0.004, 0.001 and 0.01) respectively. Again there was no oedema inhibition by indomethacin at the later time-points.When comparing the calculated areas under the curve «increase in paw volume - time¼ we found no differences between the values of control and study groups: naproxen (15 mg/kg) and indomethacin (10 mg/kg). We think that these values of areas under the curve «increase in paw volume - time¼ represent the total inflammatory reaction induced by carrageenan and need to be used for the assessment of future potential anti-inflammatory agents which should not only produce short-term symptomatic oedema suppression, but change the nature of the oedema response, potentially with alternative mechanisms of action. Our experimental findings are in accordance with the well-known lack of effects of NSAIDs on the outcomes of chronic inflammatory diseases [12]. This may be due to the fact that they suppress the development and symptoms of inflammation at the early stages, but the reaction to inflammatory stimuli develops fully over the longer period of time and takes its full course nonetheless. This proves that traditional modeling approaches to future potential anti-inflammatory agents development needs re-assessment. CONCLUSIONS: Single-dose administration of naproxen (15 mg/kg) or indomethacin (10 mg/kg) exerts decrease in rat paw oedema volume at no later than 5 hours after oedema induction by carrageenan. Evaluating anti-inflammatory activity by the areas under the curve «increase in paw volume - time¼ proves that a single-dose NSAID's administration has no effect on the inflammatory response when evaluated not by single time-point index (at 3 or 5 hours), but by assessing the oedema development and duration over 192 hours (8 days).

[MODELING INFLAMMATORY EDEMA: ARE THE MODELS INTERCHANGEABLE].

Experimental modeling of inflammatory edema by sub-plantar injection of carrageenan and formalin in mice and rats is widely used to evaluate potential anti-inflammatory activity of new drugs. This systematic analysis of published data showed that carrageenan induced paw edema model is used for evaluating the anti-inflammatory activity mostly in rats rather than mice. Formalin induced paw edema in rats and mice is used primarily for evaluation of the analgesic activity of drugs. Taken together, the results of this systematic review of available literature on edema modeling substantiate recommendation to use carrageenan paw edema in rats and formalin paw edema in mice as complementary, but not interchangeable models of inflammation.

[Experimental comparative study of the analgesic activity of the diurnal anxiolytic mebicar, amitriptyline, and diazepam].

It is established that mebicar, amitriptyline, and diazepam administered intraperitoneally at therapeutic doses increase the pain thresholds in mice as manifested in "hot plate" analgesia test. Mebicar was more effective than diazepam and not inferior to amitriptyline in increasing the pain thresholds at earlier time points.

[Anti-inflammatory properties of antidepressants (a review of experimental results)].

The review presents comparative description of anti-inflammatory effects of antidepressants from various classes studied on carrageenan- and formalin-induced paw edema in rats. The role of the central and peripheral components in the mechanisms of anti-inflammatory activity and the drug effects on production of mediators and modulators of inflammation is discussed.

[Dimephosphone shows anti-inflammatory and anti-oxidative activity on chronic autoimmune inflammation model].

The anti-inflammatory activity of dimephosphone, xydiphone (ethidronate), and ionol (dibunol) and their effects on lipid peroxidation (LPO) indices and the activity of antioxidant enzymes in blood were studied on the model of Freund adjuvant induced arthritis in white laboratory rats. The Freund adjuvant induced chronic arthritis and increased the concentrations of PLO products and nitrite ions in the blood plasma. Only dimephosphone showed an anti-inflammatory action. Dimephosphone and ionol inhibited the LPO, whereas xydiphone did not influence the LPO indices in the blood.

Contractile activity of human greater saphenous vein mediated by P2-receptors.

In vitro experiments were carried out to measure the contractile responses to P(2)-receptor agonists in the greater saphenous vein isolated from patients with obliterating vascular atherosclerosis and varicose veins of the legs. In patients with varicose veins, the contractile responses of the greater saphenous vein to ATP, alpha,beta-methylene-ATP, and UTP were significantly lower than in patients with obliterating atherosclerosis, while the responses to ADP, adenosine, and 2-methylthio-ATP were similar in both groups. These data attest to the presence of P(2)-receptor-mediated contraction component in the greater saphenous vein, which are pronouncedly weakened during varicose disease.

[New approaches to prognosis of risk of development of gastropathies induced by nonsteroid anti-inflammatory agents]. / Novye podkhody k prognozirovaniiu riska razvitiia gastropatii, indutsirovannykh nesteroidnymi protivovospalitel'nymi sredstvami.

A pharmacological test with indomethacin is suggested for predicting the risk of NSAID-induced gastropathy. It is shown that patients with diagnoses of rheumatoid arthritis and osteoarthrosis can be divided into two groups with respect to the indomethacin test--"vulnerable" and "resistant", characterized by a high and low risk of NAIDS-induced gastropathy development, respectively. The proposed index of resistance to this disorder, together with the glutathione redox buffer test index, can be used as reliable criteria for predicting the NSAID-induced gastropathy.