Banu Coskun

The effects of caffeic acid phenethyl ester on tissue damage in lung after hindlimb ischemia-reperfusion

AIM The aim of this study was to investigate the effects of caffeic acid phenethyl ester (CAPE) on the lungs as a remote organ after performing hindlimb ischemia-reperfusion (I/R) and by assessing biochemical and histopathological analysis. METHODS The animals were divided into three groups: control, I/R, and I/R with CAPE. I/R period for 8 h was performed on the right hindlimb of all the anesthesied rats in I/R and CAPE with I/R group. In the CAPE with I/R group, the animals received CAPE 10 microM by intraperitoneal injection 1h before the reperfusion. The animals in the control and I/R groups received a similar volume of saline solution by means of intraperitoneal injection. At the end of the reperfusion period, a midsternotomy was performed. Blood, bronchoalveolar lavage (BAL) and lung tissue were obtained, and were used for biochemical and histopathological examination. RESULTS The tissue and serum malondyaldehyde levels were significantly lower in the control (P=0.0001 and 0.001, respectively) and in the CAPE with I/R groups (P=0.0001 and 0.003, respectively) compared to the I/R group. Tissue Na(+)-K(+) ATPase activity in the CAPE with I/R group was significantly higher than in the I/R group (P=0.0001). Reduced activity was found in the I/R group compared to the control group (P=0.0001). Myeloperoxidase activity (P=0.001) and protein concentration (P=0.034) in BAL were significantly reduced in CAPE-treated animals when compared with the I/R group. A decreased activity and protein concentration were found in the control group compared to the I/R group (P=0.0001 and 0.024, respectively). The lungs of the I/R group displayed intense peribronchial and perivascular leukocytic infiltration in histopathological examination compared to the CAPE with I/R group (P<0.05). CONCLUSION CAPE seems to be effective in protecting remote organ injury caused by increased oxidative stress and neutrophil accumulation that results from an I/R injury.

No effect of GA-AS (904 nm) laser irradiation on the intact skin of the injured rat sciatic nerve

AbstractWe evaluated the electrophysiological and histopathological effects of low-energy gallium arsenide (904 nm) laser irradiation on the intact skin injured rat sciatic nerve. Twenty-four male Wistar rats were divided into three groups (n=8 each). At the level of proximal third of the femur the sciatic nerve was crushed bilaterally with an aneursym clip (Aesculap FE 751, Tuttingen, Germany) for half a second. A gallium arsenide laser (wavelength 904 nm, pulse duration 220 ns, peak power per pulse 27 W, spot size 0.28 cm2, pulse repetition rate 16, 128 and 1000 Hz; total applied energy density 0.31, 2.48 and 19 J/cm2) was applied to the right sciatic nerve for 15 min daily at the same time on 7 consecutive days. The same procedure was performed on the left sciatic nerve of same animal, but without radiation emission, and this was accepted as control. Compound muscle action potentials were recorded from right and left sides in all three groups before surgery, just at the end of injury, at the 24th hour and on the 14th and 21st days of injury in all rats using a BIOPAC MP 100 Acquisition System Version 3.5.7 (Santa Barbara, USA). BIOPAC Acknowledge Analysis Software (ACK 100 W) was used to measure CMAP amplitude, area, proximal and distal latency, total duration and conduction velocity. Twenty-one days after injury, the rats were sacrificed. The sciatic nerves of the operated parts were harvested from the right and left sides. Histopathological evaluation was performed by light microscopy. Statistical evaluation was done using analysis of variance for two factors (right and left sides) repeated-measures (CMAP variables within groups) and the Tukey–Kramer Honestly Significant Difference test (CMAP variables between laser groups). The significance was set at p ≤ 0.05. No statistically significant difference (p ≥ 0.05) was found regarding the amplitude, area, duration and conduction velocity of CMAP for each applied dose (0.31, 2.48 and 19 J/cm2) on the irradiated (right) side and the control (left) side, or between irradiated groups. Twenty-one days after injury there were no qualitative differences in the morphological pattern of the regenerated nerve fibres in either irradiated (0.31, 2.48 and 19 J/cm2) or control nerves when evaluated by light microscopy. This study showed that low-energy GaAs irradiation did not have any effect on the injured rat sciatic nerve.

Alcohol-induced lung damage and increased oxidative stress.

Background: Alcohol-induced lung damage may be associated with increased oxidative stress. Objective: Our aim was to investigate alcohol-induced changes in the biochemistry and histopathology of the lung. Methods: Rats were divided into two groups, a control group and an ethanol group. The ethanol group received 2 g/kg ethanol (total: 3 ml) intraperitoneally. The controls were given the same amount of saline via the same route. Three hours later, the rats were sacrificed, and blood and lung tissue samples were obtained. Oxidative stress was assessed by measuring the levels of erythrocyte reduced glutathione (GSH), tissue malondialdehyde (MDA), myeloperoxidase (MPO) and Na+-K+ ATPase. Histopathologic evaluation of the lung tissues was also performed. Results: In the ethanol group, serum and tissue MDA levels and MPO activities were increased (p = 0.007, p = 0.001 and p = 0.000), and lung tissue Na+-K+ ATPase activities and erythrocyte GSH were decreased (p = 0.001 and p = 0.000) compared to the controls. Histopathologic examination demonstrated alveolocapillary thickening, alveolar degeneration, leukocyte infiltration and erythrocyte extravasation in the lungs of the ethanol group (p < 0.05). Conclusion: These results suggest that high-doseacute alcohol administration aggravates systemic and local oxidative stress leading to acute lung injury, ranging from mild pulmonary dysfunction to severe lung injury. It should be borne in mind that rapid onset of the acute respiratory distress syndrome (ARDS) may also be due to increased oxidative stress following alcohol abuse, especially when ischemic disturbances, e.g. coronary heart disease, acute ischemia of the extremities and traumatic accidents, are concomitantly present. Therefore, precautions against ARDS may prevent morbidity and mortality in alcohol-induced lung damage in at-risk patients.

Effects of trimetazidine on tissue damage in kidney after hindlimb ischemia-reperfusion.

The objective of this study was to investigate the effects of trimetazidine (TMZ) on tissue damage in kidney after hindlimb ischemia-reperfusion (I/R), by assessing blood biochemical assay and histopathological analysis. Adult male Wistar rats were divided into two groups. TMZ 10 mg kg(-1)day(-1) was administrated twice a day for 10 days to the treatment group (group T, n=10). Sham group was given only 5% gum arabic (group S, n=10). On 11th day of treatment, 8h I/R period was performed on right hindlimb of the rats. At the end of reperfusion period, a 5 ml blood withdrawn from ascending aorta for biochemical assays and their right kidneys were harvested for histopathological examination. Superoxide dismutase, Na(+)-K(+) ATPase, and reduced glutathione levels were significantly increased in group T (P<0.001). On the other hand, myeloperoxidase and malondialdehyde levels were significantly less in group T than group S (P<0.001). Kidneys from the sham-operated group displayed intense leukocytic infiltration in histopathological examination. These overall results strongly suggested that TMZ contributes renal protection from hindlimb I/R injury by decreasing systemic oxidative stress.

Effects of Daflon on oxidative stress induced by hindlimb ischemia/reperfusion.

The objective of this study was to investigate the effects of Daflon 500 mg on tissue damage in kidney after ischemia/reperfusion hindlimb, by assessing blood biochemical assay and histopathological analysis. Rats were given Daflon 80 mg x kg(-1) x day(-1) for 10 days. On 11th day of treatment, 4h ischemia followed by 4 h reperfusion period was performed on right hind limb of the rats. Control groups were given only arabic gum and were subjected to same ischemia/reperfusion period. At the end of reperfusion period, erythrocyte superoxide dismutase, Na(+)-K(+) ATPase and reduced glutathione levels were increased in the rats erythrocytes in Daflon group (P<0.01, for all). On the other hand, serum myeloperoxidase and malondialdehyde levels were significantly lower in the Daflon-received rats (P<0.01, for all). Histopathological studies demonstrated that, there was a prominent tubulointerstitial injury with loss of brush border and this degeneration was accompanied by segmental glomerular degeneration also for both control and Daflon group. Daflon-received group animals displayed significantly less periglomerular and perivascular leukocytic infiltration (P=0.015). These overall results suggest that Daflon contributes renal protection from hind limb ischemia/reperfusion injury in some degree, by decreasing systemic oxidative stress.

ELECTROPHYSIOLOGICAL AND HISTOPATHOLOGICAL EVALUATION OF RESPIRATORY TRACT, DIAPHRAGM, AND PHRENIC NERVE AFTER DICHLORVOS INHALATION IN RATS

The aim of this study was to investigate the dose-related effects of dichlorvos inhalation on electrophysiological alterations of diaphragm and phrenic nerve and the changes in the histologic structure of respiratory system. This study was performed on 33 rats divided into 5 groups, inhaling 1, 2, 5, 10, and 15 µg/L of dichlorvos, respectively. Electrodiagnostic investigations of diaphragm and phrenic nerve were made before and after inhalations. Aspiration samples were taken from lungs to evaluate the presence of infection agents. The airways, lungs, and diaphragms were dissected out for histologic investigation. Rats exposed to a low concentration of dichlorvos (1-5 µg/L) showed no symptoms of intoxication, but exposure to higher doses (10-15 µg/L) induced dyspnea in several animals. Lower doses of dichlorvos revealed no electromyographic changes on diaphragm, whereas higher doses revealed a clear neuropathic involvement. Delayed phrenic nerve motor conduction velocity was noted for each group (p < .05). Morphologic changes on the tracheal epithelium, hyperplasia, thickening of the blood-air barrier, degeneration in alveoli, and ductus alveolaris were seen in histopathologic investigation. In conclusion, the acute inhalation of dichlorvos caused clear evidence of neuropathic involvement of the diaphragm and the phrenic nerve. Also, toxic pneumonitis and injury to the tracheal epithelial were noticed.

Addition of Fusidic Acid Impregnated Bone Cement to Systemic Teicoplanin Therapy in the Treatment of Rat Osteomyelitis

Abstract We compared the efficacy of the combination of fusidic acid impregnated bone cement and systemic teicoplanin to systemic teicoplanin alone in implant-related osteomyelitis model in the rats. Foreign bodies were implanted into the medullary channels of 30 rat tibias after intramedullary inoculation of methicillin-resistant Staphylococcus aureus. Following proof of induction of osteomyelitis in the rats on the 21st day, a bone cement rod including 1/40 ratio of fusidic acid was inserted into the medullary channel of the tibias in the study group. Teicoplanin was administered i.m. at 20 mg/kg/day for 14 days to both the study and control groups. At the end of the treatment, the tibias were examined macroscopically, microbiologically and histopathologically. The elimination rate with the teicoplanin+fusidic acid combination was 81.8%, while with teicoplanin alone was 55.6% (p=0.33). Although the difference between the two groups was not statistically significant, the combination treatment had a positive effect in eliminating the microorganism.

Evaluation of the toxic effects of cypermethrin inhalation on the frog heart.

Cypermethrin is widely used as an insecticide on animals and in agriculture, the home, and the garden. The effect of inhaled cypermethrin on the cardiac mechanics, electrophysiology, and ultrastructure in frogs was investigated in this study. Four groups received 100 microL of cypermethrin via inhalation for different exposure times, and one group was used as a control. Electrical and mechanical activities of the heart were recorded, and heart samples were examined at light and transmission electron microscopic levels for all groups. The atrial and ventricular contractile forces on the mechanogram, the amplitude of the P wave and the QRS complex on electrocardiogram, and the heart rate were significantly decreased in cypermethrin-inhalated frogs. The total duration of contraction was prolonged in the study groups. Ultrastructurally, dilatation in smooth endoplasmic reticulum cisterns, a decrease in the number of mitochondria, disorganization in the myofibrils of myocytes, and necrotic changes in endothelial cells were observed. These results suggest that cypermethrin has cardiotoxic effects that increase with exposure time.

Interferon-α2b may impair myelinization of rat optic nerve

This story investigated the effects of interferon-alpha-2b (IFN-α2b) on the optic nerves of 17 adult male Wistar albino rats. Animals were divided into 3 groups: 6 rats (group 1) received 7.5 units (5 mIU/m2) IFN-α2b—a normal treatment dose, and 6 (groups 2) received 30.0 units (20 mIU/m2)—a high dose; 5 rats (control group) received 0.5 mL saline. Test substances were delivered by intraperitoneal injection 3 times a week for 3 weeks with animals under inhalation anesthesia. After the rats were sacrificed, their optic nerves were dissected, sectioned, and examined under an electron microscope. The mean thicknesses of the basal membranes of blood vessels were 86.354 nm in the control group, 104.297 nm in group 1, and 140.181 nm in group 2. Basal membrane changes in IFN groups were dose dependent. Mitochondrial swelling, degeneration, increased diameter of vacuoles, and vacuolization in the cytoplasm of oligodendrocytes and astrocytes were also observed. IFN-α2b has histopathologic effects on blood vessels and cells of the optic nerve.

Combined Therapy of Teicoplanin and Caffeic Acid Phenethyl Ester (CAPE) in the Treatment of Experimental Mediastinitis in the Rat

Abstract Post-sternotomy mediastinitis affects 1-3% of patients undergoing cardiac surgery and is lethal in 10-47% of these patients. We investigated the effect of an antioxidant/anti-inflammatory agent, caffeic acid phenethyl ester (CAPE), in the attenuation of inflammatory response induced by methicillin-resistant Staphylococcus aureus (MRSA) infection in a rat experimental mediastinitis model. Rats, divided into six equal groups, received MRSA precolonized stainless steel wire pieces implanted into their mediastinal spaces. Control group and CAPE control group received saline and CAPE 10 μmol/kg.day−1 respectively, where Group A received a single dose of teicoplanin 24 mg/kg i.m. for the first day and then 12 mg/kg.day−1. Group B received teicoplanin as in Group A plus CAPE 10 μmol/kg. day−1 intra-peritoneally. Group C received teicoplanin 60 mg/kg i.m. for the first day and then 30 mg/kg.day−1 and Group D received teicoplanin as in Group C plus CAPE 10 μmol/kg.day−1. By the end of 14 days rats were sacrificed and serum malondialdehyde (MDA), myeloperoxidase (MPO), nitric oxide (NO), urea and creatinine levels were evaluated. Mediastinal organ tissues were collected for histopathological analysis. Infection rates in all the drug-treated groups were lower than the control groups (P = 0.002) but statistical significance was attained only between the groups A and D (P = 0.018). In connective tissues and the peribronchial area polymorphonuclear leukocytic (PNL) infiltration in the treatment groups, although becoming very close, did not reach statistical significance (P = 0.053, P = 0.075, respectively). PNL infiltration especially in the peribronchial tissues of the Group B animals was found to be significantly less than the Control and CAPE Control groups with P values of 0.013 and 0.010, respectively. MDA and MPO levels were significantly lower in the treatment groups (P < 0.001 and P < 0.001 respectively). Levels of the degradation products of NO were lower in treatment groups compared to two control groups (P = 0.003, P = 0.005). NO levels in Group D were lowest among all treatment groups (P = 0.001). It has been demonstrated that although bacterial colonization can be controlled in mediastinitis, the inflammatory response persists. The combination of an antioxidant / anti-inflammatory agent, CAPE, added to standard antibiotic therapy might be effective in the treatment of post-sternotomy mediastinitis due to MRSA.