Ljiljana Gojkovic-Bukarica

Effect of potassium channel opener pinacidil on the contractions elicited electrically or by noradrenaline in the human radial artery.

In order to discover an agent that can prevent spasm of the human radial artery, the aim of our study was to evaluate the effect of the K(+) channel opener, pinacidil, on contractions in the radial artery. Contractions of the radial artery were evoked by exogenously applied noradrenaline or by electrical field stimulation (EFS, 20Hz, neurogenic). Pinacidil induced concentration-dependent inhibition of both EFS- and noradrenaline-evoked contractions of the radial artery. Glibenclamide, a selective blocker of ATP-sensitive K(+) channels (Kir6.x containing subunit) antagonized in the same manner the pinacidil-induced inhibition of neurogenic contractions and contractions evoked by exogenous noradrenaline. The inhibition of pinacidil relaxation by tetraethylammonium (TEA), a blocker of Ca-sensitive K(+) (K(Ca)) channels, was more pronounced in EFS-contracted preparations. A blocker of voltage-sensitive K(+) (K(V)) channels, 4-aminopyridine (4-AP), inhibited pinacidil relaxation only in EFS-contracted preparations. In order to test the presence of different K(+) channels, immunohistochemistry of K(+) channels expression in the radial artery was performed. The vascular wall of the human radial artery showed variable positivity with the following applied antibodies: Kv1.2, Kv1.3, Kir6.1, and K(Ca)1.1. The antibodies against Kv1.6, Kv2.1, and Kir6.2 channel subunits were completely negative. These results suggest that the inhibitory effect of pinacidil on contractions of the human radial artery might be postsynaptic and associated with opening of smooth muscle Kir6.1-containing K(ATP) channels. TEA- and 4-AP-sensitive K(+) channels may also contribute to pinacidil effect in the human radial artery.

Effect of Wine Polyphenol Resveratrol on the Contractions Elicited Electrically or by Norepinephrine in the Rat Portal Vein

We investigated the effects of resveratrol on rat portal vein (RPV) contractility without endothelium. Contractions were produced by electrical field stimulation of perivascular nerves (EFS), norepinephrine (NE), adenosine 5′‐triphosphate (ATP), high K+ solution and by calcium chloride (CaCl2) in Ca2+‐free and high K+, Ca2+‐free solution. The EFS‐evoked contractions were more sensitive to resveratrol and to NS1619‐selective openers of big calcium‐sensitive (BKCa) channels, than NE‐evoked contractions. Effects of resveratrol on the ATP‐evoked contractions were weak. Blockers of BKCa channels partly inhibited the effect of resveratrol only in EFS‐contracted preparations. Western blotting showed that RPV expressed KCa1.1 protein. Inhibitors of ATP‐ and voltage‐sensitive K+ channels did not modify the effects of resveratrol. None of the antagonists of K+ channels affected the resveratrol inhibition of NE‐evoked contractions and effect of high concentrations of resveratrol on the EFS‐evoked contractions. Resveratrol more potently inhibited CaCl2 than potassium chloride contractions of RPV. Thus, BKCa channels partly mediate the inhibitory effect of resveratrol on the neurogenic contractions of RPV. The smooth muscle Ca2+ channels and/or Ca2+ mobilizing through cells might be involved in the effects of resveratrol on the contractility of RPV. Our results are important for better understanding the impact of resveratrol on the portal circulation. Copyright

The effects of potassium channel opener P1075 on the human saphenous vein and human internal mammary artery.

Because adrenergic contractions can contribute to the development of life-threatening spasm of coronary artery bypass graft, this study was performed to investigate the effect of adenosine 3-phosphate (ATP)-sensitive K+ channel (KATP) opener P1075 on contractions of isolated human saphenous vein (HSV) and human internal mammary artery (HIMA). Phasic contractions were evoked by electric field stimulation (20 Hz) and noradrenaline. The sustained contractions were evoked by phenylephrine. The presence of pore-forming Kir6.1 and Kir6.2 subunits of the KATP channels in the HIMA and only Kir6.2 in the HSV was confirmed immunomorphologically. P1075 inhibited in the HSV only, the electrical field stimulation contractions more strongly than noradrenaline contractions. In addition, the phenylephrine contractions of HSV were more sensitive to P1075 in comparison to those of HIMA. Glibenclamide, a KATP channel blocker antagonized the vasodilatation produced by P1075 in both grafts differently, because its effect was more prominent on the P1075-induced inhibition of contractions of HSV than of HIMA. We conclude that P1075 has a vasorelaxant effect and inhibited adrenergic contractions of the tested grafts. This effect is graft and vasoconstrictor selective and seems to be mediated by Kir6.1- and/or Kir6.2-containing KATP channels. Thus, P1075 can be considered as a potential drug in the prevention of graft spasm.

The differential effect of resveratrol on the renal artery of normal and diabetic rats

Results Resveratrol relaxed RA of normal rats more potently than RA of rats with diabetes (EC50 8 and 50 μM, respectively). L-NAME and methylene blue partly antagonized the relaxation of RA of normal animals only. A nonselective blocker of voltage-gated K (KV) channels, 4-aminopyridine (4-AP) partly inhibited the relaxation of RA of normal as well as of diabetic rats. However, margatoxin, a selective antagonist of KV1.x channels, completely antagonized the relaxation of RA of diabetic rats only. Glibenclamide, a highly selective blocker of ATP-sensitive K channels, did not block resveratrolinduced relaxation in both experimental models. Conclusions In conclusion, we have shown that resveratrol induces a strong endothelium-dependent relaxation of RA of normal rats, and that 4-AP-sensitive K channels are involved in this relaxation. In diabetic rats, resveratrol induced NO-independent relaxation and maragtoxinsensitive K channels are involved.

Improving the low solubility of resveratrol

Background Resveratrol, a polyphenol mainly present in grapes and red wine, demonstrated interesting biomedical properties for its cardioprotective action due to inhibition of the oxidation of low-density lipoprotein (LDL) and of platelet aggregation, inhibitory effects on cancer promotion and propagation and anti-inflammatory activities. These potential therapeutic and prophylactic applications are limited by the low bioavailability caused by its physical properties. Additionally, resveratrol has low water solubility and stability making its clinical success a formidable technological and medical challenge. The aim of this work is to present results of improvement of solubility of resveratrol through micellar and liposomal incorporation.

The effects of resveratrol on rat behaviour in the forced swim test

INTRODUCTION The trans-isomer of resveratrol is the active ingredient of Poligonum cuspidatum, known for its medicinal properties and traditionally used in the treatment of neuropsychiatric disorders. It is also found abundantly in the skin of red grapes and red wine. Previous studies have suggested that trans-resveratrol demonstrates a variety of pharmacological activities including antioxidant, anti-inflammatory, as well as neuroprotective properties and procognitive effects. OBJECTIVE The goal of the present study was to examine the influence of trans-resveratrol on behavior in rats and its antidepressant properties. METHODS Male Wistar rats were treated intraperitoneally (i.p.) with the increasing doses of trans-resveratrol (5, 10 and 20 mg/kg) or vehicle (dimethyl sulfoxide--DMSO), 30 minutes before testing of the spontaneous locomotor activity or forced swimming. For the experiments, the behavior of the animals was recorded by a digital camera, and the data were analyzed by one-way ANOVA, followed by Tukey post-hoc test. RESULTS Testing of spontaneous locomotor activity, after the application of vehicle or increasing doses of trans-resveratrol, showed no statistically significant difference between groups (p > 0.05). In the forced swim test, one-way ANOVA indicated statistically significant effects of trans-resveratrol (p < 0.001).Tukey post-hoc test showed that resveratrol significantly decreased immobility time at the doses of 10 mg/kg and 20 mg/kg, manifesting the acute antidepressant-like effects. There were no statistically significant differences between the resveratrol treatment of 5 mg/kg and vehicle (p > 0.05). CONCLUSION The results from our study suggest that transresveratrol produces significant effects in the central nervous system. After single application, it has acute antidepressant effects, but without influence on locomotor activity.

The Novel Phenylpropiophenone Derivates Induced Relaxation of Isolated Rat Aorta

Our aim was to define how different chemical properties of newly developed phenylpropiophenone derivates (PhPds) influenced their potency and efficacy to relax rat aorta. A contribution of ion channels in the PhPds and propafenone mechanism of vasodilatation was tested. PhPds were syntethysed by substitution in the benzyl moiety with -F, -CH3 or -CF3 groups on the ortho or para position. The vasodilatation by PhPds was examined on the rings of rat aorta precontracted with phenylephrine. In order to test involvement of voltage-gated Na+ and K+ channels and L-type Ca2+ channels in a mechanism of action of PhPds, we used their blockers: lidocaine, nifedipine and 4-aminopiridine, respectively. Aorta was more sensitive to 5-ortho-trifluoromethyl derivate than to propafenone and other PhPds. The 5-para-methyl derivate had lower potency and efficacy than propafenone and other PhPds. Lidocaine did not influenced relaxation induced by PhPds, but slightly inhibited the effect of propafenone. The 4-aminopiridine only inhibited relaxation induced by 5-para-methyl derivate. Nifedipine inhibited relaxation of the rat aorta induced by 5-ortho-trifluoromethyl derivate and by propafenone. Introduction of 5-ortho-trifluoromethyl and 5-para-methyl group in the benzyl moiety of propafenone molecule changed its potency, efficacy and mechanism of action in the rat aorta. The 4-aminopiridine- and nifedipine sensitive ion channels are involved in mechanism of action of 5-para-methyl and 5-ortho-trifluoromethyl derivate. The introduction of other tested groups in the benzyl moiety does not affect pharmacological properties of the PhPds in relation to propafenone.

Quantification of the acute effect of a low dose of red wine by nonlinear measures of RR and QT interval series in healthy subjects

The measures of nonlinear properties of RR interval and QT interval time series are sensitive to physiologically- or pathologically-induced complexity/regularity changes, but were not used to estimate the effect of alcohol intake. We wanted to examine the potential of these measures to quantify the acute effect of a low dose of red wine in healthy subjects. In separate experiments, fourteen young volunteers drank 200ml of red wine and a control drink with equal concentration of ethanol. ECG in supine position was recorded 20min before and 60min after drink intake. RR interval and QT interval series were extracted from ECG and we calculated variability, scaling exponents (α1 and α2) and sample entropy (SampEn) for both series. Systolic and diastolic blood pressures (BP) were measured every 10min. The immediate effect of both the drinks was equal: HR, BP and QT variability exhibited a sudden increase and then a decrease. However, the prolonged effect of wine and the control drink was different. Wine decreased both BP (p<0.05) and reduced complexity of RR and QT series (increased scaling exponents and decreased SampEn). The control drink prolonged QT and RR intervals (p<0.05). These results point out that the nonlinear properties of RR and QT interval series could be used to differentiate the effect of wine and ethanol. Changes in RR and QT interval series induced by a low dose of red wine are more detectable by methods that quantify the structure of the series than by methods that quantify their variability.

Spontaneous contractions of isolated rat portal vein under temperature perturbations

We studied nonlinear dynamics underlying spontaneous rhythmical contractions of isolated rat portal vein. The signals were acquired at four different temperatures important in isolated blood vessels preparations: 4, 22, 37 and 40°C. To characterize the system’s nonlinearity, we calculated the largest Lyapunov exponent, sample entropy and scaling exponents. Evidence for nonlinearity was provided by analysis of surrogate data generated from the phase-randomized Fourier transform of the original sequences. Positive values of the largest Lyapunov exponent were obtained for the time series recorded under applied conditions, indicating that the system preserves its chaotic deterministic nature even far from the physiological temperature range. Scaling exponents revealed three distinctive regions with different correlation properties. The calculated measures that characterize the time series obtained at 4°C were significantly different from those derived from data obtained at higher temperatures. System’s dynamics becomes more complex or less predictable as temperature approaches physiological value. The computation of the largest Lyapunov exponent, sample entropy and correlation measures gave an insight into the complex dynamics of the isolated blood vessels rhythmicity. We identified different modes of rhythmical contractions of isolated rat portal vein which could improve understanding of possible control mechanisms in vivo.

Differences in antimicrobial consumption, prescribing and isolation rate of multidrug resistant Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii on surgical and medical wards

In order to provide guidance data for clinically rational use of an antibiotics consuption, prescribing and prevalence of multidrug resistant (MDR) Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii were monitored on the surgical (S) and medical (M) wards of the University Hospital Center “Dr. Dragisa Misovic-Dedinje” (Belgrade, Serbia), in the study period from 2012 to 2015. Appropriateness of antimicrobial use was evaluated using the Global-Prevalence Survey method designed by the University of Antwerp. The percentages of MDR pathogens relative to the total number of isolates of K. pneumoniae and P. aeruginosa were higher on the S (86.2% and 49.1%) than on the M (63.2% and 36.9%) wards. The percentage of MDR A. baumannii was not different between S (93.7%) and M (79.5%) wards. An overall antibiotics consumption (defined daily doses/100 bed-days) during study was 369.7 and 261.5 on the S and M wards, respectively. A total of 225 prescriptions of antimicrobials were evaluated in138 adults admitted to wards on the day of the survey. The percentage of antimicrobials prescribed for prophylaxis on the M and S wards were 0% and 25%, respectively. Therapies were more frequently empiric (S, 86.8% and M, 80%). The percentages of medical errors on the S and M wards were 74.6% and 27.3%, respectively. The quality indicators for antibiotic prescribing on the S and M wards were as follows: the incorrect choice of antimicrobials (35.6% vs. 20.0%), inappropriate dose interval (70.6% vs. 16.9%) or duration of therapy (72.5% vs. 23.1%), a non-documented stop/review data (73.6% vs. 16.9%) and divergence from guidelines (71.9% vs. 23.1%). Treatment based on biomarkers was more common on the M wards as compared to the S wards. The increasing prevalence of MDR pathogens, a very high consumption and incorrect prescribing of antimicrobials need special attention, particularly on the S wards.