Ischaemic postconditioning reduces serum and tubular TNF-α expression in ischaemic-reperfused kidney in healthy rats.

OBJECTIVE: We studied the protective effects of postconditioning (PS) in healthy and hypercholesterolemic rats after renal ischaemia-reperfusion (IR) injury. We aimed to examine cytokine expression and apoptosis in tissue damage after revascularisation (TNF-α levels in serum and tissue). METHODS: Male Wistar rats (n = 32) were divided into four groups. The animals of normal feed groups (NF) were fed with normal rat chow and the cholesterol feed groups (CF) were fed with 1.5% cholesterol containing diet for 8 weeks. Anaesthetized rats underwent a 45-min cross-clamping in both kidney pedicles. Ischaemia was followed by 120-min reperfusion with or without PS protocol (group PS vs. IR). Postconditioning was induced by four intermittent periods of ischaemia-reperfusion of 15-s duration each. Serum cholesterol, triglyceride, urea and creatinine levels were determined. Proinflammation was characterized by the measurement of serum TNF-α. Tissue injury in kidney was determined by formaline-fixed, paraffin-embedded tissue sections. Tissue TNF-α levels were determined by immunohistochemistry. RESULTS: Significant elevation was observed in serum TNF-α level after IR injury in normal feed groups, which was reduced by PS. In CF group neither the elevation nor the postconditioning induced reduction were as significant as in the NF groups. In normal feed group PS caused a significant reduction in tissue TNF-α level which was significantly higher in CF. CONCLUSIONS: Ischaemic postconditioning proved to be an effective defense against IR in NF groups, but it was ineffective in CF groups in kidney tissue.

Effect of vitamin E on reperfusion injuries during reconstructive vascular operations on lower limbs.

INTRODUCTION: The challenge against reperfusion injury and tissue oxidative stress, especially in vascular surgical interventions has an essential importance to reach the optimal clinical result. Numerous experimental attempts have proved the positive antioxidant effect of vitamin E in both chronic and acute phase models. In our study we monitored the effect of continuous preoperative treatment with vitamin E, on oxidative stress and tissue inflammation reactions developed after reconstructive operations. PATIENTS AND METHODS: 32 patients have been involved in a randomized, prospective study, all suffering from AFS occlusion proved by angiography, and all undergone supragenual reconstruction. Duration of ischemia and amount of tissues under vascular clamping were almost the same in all patients. In the group treated with E-vitamin, we administered 1 x 200 mg of vitamin E p/o from the preoperative day till the 7th post operative day. Patients of the second group did not receive vitamin E. MATERIALS AND METHODS: Peripheral blood samples were collected immediately before operation and at the end of the second reperfusion hour (early reperfusion period). Late reperfusion period has been monitored by analyzing blood samples taken at 24th hour and 7th day next to the operative ischemia. Among oxidative stress parameters, direct measurement of reactive oxygen intermediator (ROI) and determination of antioxidant state (GSH, Total-SH group, SOD) have been performed. Malondialdehyde was chosen as marker for lipidperoxidation. Inflammation reactions were monitored up on expression of adhesion molecules (CD11a and CD18). We also controlled the oscillation of myeloperoxidase (MPO) activity. RESULTS: Our study has proved that preoperative (from the preoperative day till the 7th post operative day) administration of 200 mg vitamin E could reduce the level of oxidative stress developed after ischemic-reperfusion insult (lipidproxidation, antioxidant enzymes). According to our results, the prooxidant-antioxidant imbalance also diminished in the group with E-vitamin treatment. We proved that elective administration of vitamin E could decrease the WBC activity (MPO activity, free radicals production, expression of adhesion molecules) and its consequential local inflammation process, during early reperfusion.

Pathology and diagnostic options of lower limb compartment syndrome.

BACKGROUND: The indication of surgical treatment in lower limb compartment syndrome mostly depends on the clinical signs which can be often uncertain, resulting in delayed insufficient intervention. AIM: The aim of the study was to evaluate the progression of compartment syndrome by measuring of intracompartmental pressure and monitoring of decreased tissular oxygenation, indicating an insufficient secondary microcirculation. MATERIALS AND METHODS: 16 patients were examined in our study (12 males, 4 females, mean age: 62.7+/-9.5 years), who underwent acute lower limb revascularization surgery for a critical (lasting more than 4 hours) limb ischemia. The indications were: 5 iliac artery embolizations and 11 femoral artery occlusions. After revascularization, on the second postoperative day, we detected significant lower limb edema and swelling of several grade. To monitor the elevated intracompartmental pressure (ICP) and to evaluate the extremital circulation, we used KODIAG pressure meter and the tissular oxygen saturation (StO2) was measured by near-infrared-spectroscopy. RESULTS: In 12 cases the ICP exceeded the critical 40 mmHg. In these patients the average StO2 was 50-53%, in spite of complete recanalization. In these cases we made urgent, semi-open fasciotomy. In 4 cases, where the clinical aspect showed compartment syndrome, the measured parameters did not indicate a surgical intervention (ICP: 25-35 mmHg, StO2: around normal). SUMMARY: A novel approach in our examination is that, besides empirical therapeutic guidelines generally applied in clinical practice, we established an objective, parameter-based ("evidence based medicine") surgical indication strategy for the lower limb compartment syndrome. Our parameter results produced by the above pressure and saturation measurements help the clinicians to decide between conservative and operative treatment of the disease.

Ischaemic postconditioning reduces peroxide formation, cytokine expression and leukocyte activation in reperfusion injury after abdominal aortic surgery in rat model.

OBJECTIVE: We studied the protective effects of ischaemic postconditioning (PS) on ischemia-reperfusion injury of the lower extremities in a rat model of abdominal aortic intervention. We aimed to examine the evoked oxidative stress, cytokine expression and leukocyte activation after revascularisation surgery. METHODS: Anesthetized animals (48 Whistar rats) underwent a 60 min infrarenal aorta cross-clamping. After the ischaemic period, an intermittent 4 times 15 s reperfusion--15 seconds ischaemic episodes--were applied (ischaemic postconditioning: group PS). Then we started a 120 min reperfusion in the aorta. In untreated group animals underwent a long ischaemia (60 min) and the following reperfusion (group IR). Peripherial blood samples were collected before operation, and in early (5, 10, 15, 30, 60 and 120 min) reperfusion periods. Serum peroxide level, TNF-alpha concentration, myeloperoxidase (MPO) activity and PMA-induced leukocyte ROS production were measured. RESULTS: In PS group, plasma peroxide level elevation was significantly lower in very early reperfusion (5-30 min) comparing to non-conditioned IR group (10.04+/-1.9 microM/l vs. 16.91+/-3.67 microM/l, p<0.05). PS also reduced serum TNF-alpha concentration (167.41+/-31.26 microg/ml vs. 116.55+/-12.04 microg/ml, p<0.05), MPO activity (1.759+/-0.239 microM/ml vs. 1.22+/-0.126 microM/ml, p<0.05) and leukocyte activation detected by PMA-induced leukocyte ROS production (5.7+/-0.96 AU/10(3) cells vs. 4.63+/-0.69 AU/10(3) cells). CONCLUSIONS: Ischaemic postconditioning could reduce ROI production after IR in early reperfusion period, thus limiting ROI mediated tissue lesion, cytokine-leukocyte activation and inflammatory responses. PS seems to be an effective tool in vascular surgery to reduce reperfusion injuries after revascularization interventions.

Reperfusion injury and inflammatory responses following acute lower limb revascularization surgery.

After revascularization of an acute arterial occlusion the development of a serious ischaemic-reperfusion injury is a menacing challenge and a hard task in peripheral vascular surgery. A whale of evidences point to oxidative stress, as an important trigger, in the complex chain of events leading to reperfusion injury. In the present study authors aimed to examine oxidative stress parameters, antioxidant-prooxidant state and leukocyte adhesion molecules (CD11a and CD18) expression following acute revascularization surgery of lower limb.10 patients were examined in the prospective randomized study. Peripheral blood sample was collected in ischaemic period, and after reperfusion in the 2nd and 24th hours, and on 7th day. Superoxide-dismutase activity, reduced glutathion concentration and leukocytes free radical production were measured. The degree of lipidperoxidation was marked with the quantity of malondialdehyde. The expressions of adhesion molecules were measured with flowcytometry.The speed and rate of free radical production significantly increased in the early reperfusion (p<0.05). The level of antioxidant enzymes decreased after revascularization. The CD11a and CD18 expression of the granulocytes significantly (p<0.05) decreased right after the revascularization, but with a gradual elevation until the 7th day they exceed the ischaemic value. Our results showed a time specific turnover of the sensitive antioxidant-prooxidant balance after revascularization operation.

Electrophysiological effects of the antiarrhythmic agent GYKI-23107 in dogs.

Cardiac electrophysiological properties of GYKI-23107, a new membrane stabilizing antiarrhythmic agent were studied in anaesthetized open-chest dogs. Epi- end endocardial electrograms (for sinus potential and for His bundle recording) were obtained during sinus rhythm and following atrial and ventricular pacing. The registration were performed under control conditions as well as five minutes after drug administration of 8 mg/kg slow i.v. or 20 minutes after 20 mg/kg intraduodenal administration respectively. GYKI-23107 did not influence significantly either the sinus cycles, PA-intervals, sinus node potentials, or the classical electrophysiological parameters of sinoatrial function as the corrected recovery time of the sinus node, sino-atrial conduction time or the secondary post-stimulation sinus cycles before and after vegetative blockade. Neither the AH intervals, anterograde Wenckebach period, nor ventriculo-atrial conduction time changed significantly. QRS duration, configuration and HV-intervals remained also unchanged after drug administration in doses which used in this study, and which seemed to be in therapeutic range. The agent did not influence significantly the effective refractory periods of the atrium and ventricle during sinus rhythm. This study suggest that the GYKI-23107 is not depressive on the anterograde (AV), retrograde (VA), intraventricular conduction and is slightly depressive on the intrinsic pacemaker properties.

Antifibrillatory effect of GYKI-23107 in induced ventricular vulnerability by local cooling and programmed stimulation in canine models.

We tested GYKI-23107 a new agent with local anaesthetic activity, in experimentally induced life-threatening ventricular arrhythmias in pentobarbitone-anaesthetized dogs. By a cooling test and programmed stimulation ventricular fibrillation was induced before and after drug administration (8 mg/kg i.v., n = 14 and 20 mg/kg i.d., n = 12). Comparative experiments were carried out with lidocaine (10 mg/kg). In this lidocaine-treated group, ventricular fibrillation could be produced at 27.7 +/- 6.6 (S.D.) min, n = 12, while after GYKI-23107 ventricular fibrillation occurred at 46.6 +/- 10.7 min, n = 14. The new compound was well absorbed from the intestines; after i.d. administration it could prevent or reduce the onset of lethal arrhythmia for more than 40 min. Its i.d. efficacy correlated well with that of i.v. administration. GYKI-23107 appears to be a safe and potent long-acting agent against ventricular dysrhythmias. It may be a promising and valuable alternative to currently available antiarrhythmic agents. The strong antifibrillatory action observed in ischaemic canine heart (n = 5) both after i.v. or i.d. administration is of special importance.

Pharmacological investigation of a new anti-arrhythmic agent, GYKI-23 107.

GYKI-23 107 is a new antiarrhythmic substance with local anaesthetic activity. Its specific pharmacological and cardiovascular effects were studied in vivo and its efficacy was compared with that of lidocaine and mexiletine. GYKI-23 107 was effective against chemically (aconitine and ouabain) induced arrhythmias after both parenteral and oral administration. In aconitine-induced arrhythmia in mice the new compound was more active than either mexiletine or lidocaine after i.p. treatment. In ouabain-induced arrhythmia in dogs, the ED50 of GYKI-23 107 was approximately half that of mexiletine after i.v. injection. GYKI-23 107 and mexiletine produced similar elevation of the fibrillation threshold in anaesthetized cats. After oral pretreatment, GYKI-23 107 showed protective effects against coronary ligation-induced arrhythmia in conscious rats. The circulatory side-effects of GYKI-23 107 in anaesthetized and conscious dogs and cats were milder then those of mexiletine. In the antiarrhythmic dose range there were no adverse cardiovascular actions.