Protective Effect of Platelet Rich Plasma on Experimental Ischemia/Reperfusion Injury in Rat Ovary.

BACKGROUND/AIMS: Ovarian torsion is a common cause of local ischemic damage, reduced follicular activity and infertility. Platelet-rich plasma (PRP) contains growth factors with demonstrated cytoprotective properties; so we evaluated PRP efficacy in a rat ischemia/reperfusion (I/R) model. METHODS: Sixty adult female Sprague-Dawley albino rats were randomly assigned to 6 groups of 8 animals each: Sham, Ischemia, I/R, Sham + PRP, I + PRP and I/R + PRP; and the remaining 12 used to prepare PRP. Ischemia groups were subjected to bilateral adnexal torsion for 3 h, while I/R and I/R + PRP groups received subsequent detorsion for 3 h. Intraperitoneal PRP was administered 30 min prior to ischemia (Ischemia + PRP) or reperfusion (I/R + PRP). RESULTS: Total oxidant status (TOS), oxidative stress index (OSI) and total ovarian histopathological scores were higher in Ischemia and I/R groups than in the Sham group (p < 0.05). PRP decreased mean TOS, OSI and histopathological scores in I + PRP and I/R + PRP groups compared to the corresponding Ischemia and I/R groups (p < 0.001). There was a strong correlation between total histopathological score and OSI (r = 0.877, p < 0.001). Peritoneal vascular endothelial growth factor was significantly higher in PRP-treated groups than corresponding untreated groups (p < 0.05). CONCLUSION: PRP is effective for the prevention of ischemia and reperfusion damage in rat ovary.

The protective effect of G-CSF on experimental ischemia/reperfusion injury in rat ovary.

OBJECTIVE: This experimental study was designed to evaluate the effect of granulocyte colony-stimulating factor (G-CSF) in ovarian ischemia and ischemia/reperfusion (I/R) injury. STUDY DESIGN: Forty-eight female adult Sprague-Dawley albino rats were divided into six groups as Group 1: sham, Group 2: torsion, Group 3: detorsion, Group 4: sham + G-CSF, Group 5: torsion + G-CSF, and Group 6: detorsion + G-CSF. Except for Groups 1 and 4, all groups underwent a dnexal torsion bilaterally for 3 h. Adnexal detorsion was applied to Groups 3 and 6 for 3 h after a 3-h torsion period. The intraperitoneal injection of G-CSF (100 IU/kg) was administered 30 min previously in Group 4, 5 and 6. At the end of the study process the animals were euthanized and their ovaries were removed for histopathological and biochemical analysis. Total oxidant status (TOS), total antioxidative status and oxidative stress index (OSI) concentrations were determined and compared. Histopathological examination of ovaries was performed for the presence of interstitial edema, congestion, hemorrhage and loss of cohesion to determine tissue damage. RESULTS: In Group 3, 4, 5 and 6, TOS, OSI and total histopathological scores of ovarian tissue were higher than in the sham group (p < 0.05). G-CSF administration decreased mean TOS and OSI levels significantly when compared with the controls (p < 0.001, p < 0.001, respectively). There was a strong correlation between the total histopathological scores of I/R injury and OSI (r = 0.862, p < 0.001). The total histopathological scores for the rats conservatively treated with G-CSF were lower than those of the control groups. CONCLUSION: G-CSF is effective for the prevention of ischemia and ischemia/reperfusion-induced damage in rat ovary.

Malondialdehyde level and adenosine deaminase activity in nasal polyps.

OBJECTIVE: Although there are many reports on adenosine deaminase (ADA) activities in different tissues, no information is available about the enzyme activity in nasal mucosa and polyp tissues. Whereas ADA is related to the production of free radicals by neutrophils, malondialdehyde (MDA) is an indicator of lipid peroxidation that is a general mechanism of tissue damage by free radicals. This study is aimed at determining and comparing the ADA activity and MDA level in nasal polyps and normal mucosa. STUDY DESIGN AND SETTING: Twenty-three patients with nasal polyps and a control group consisting of 14 patients with septal deviation and lower turbinate hypertrophy were included in the study. Tissue MDA level was measured by the method of Okawa with modification and tissue ADA activity by the method of Giusti. RESULTS: In patients with nasal polyp, mean tissue MDA level and ADA activity were 2.43 +/- 0.38 nmol/mg protein (Pr) and 0.235 +/- 0.055 U/mg Pr, respectively, which were significantly higher than those of control nasal mucosa (1.03 +/- 0.41 nmol/mg protein and 0.056 +/- 0.011 U/mg Pr, respectively) (P < 0.05). In addition, tissue MDA level was positively correlated to ADA activity in nasal polyps (r = 0.701, P < 0.001). CONCLUSIONS: The present study showed the presence of detectable ADA activity in nasal mucosa, and also significant increases in both tissue MDA level and ADA activity in NP tissue when compared to normal turbinate tissue. EBM RATING: B-2b.

Investigation of nitric oxide in pterygium.

BACKGROUND: The aim of this study was to investigate the presence of nitric oxide in pterygium. METHODS: Twenty nasal pterygium tissue samples were collected from 20 surgery patients in Kahramanmaras Sutcuimam University hospital. For control purposes, nasal conjunctivas from 10 patients undergoing limbal extracapsular cataract extraction were also collected. All specimens were preserved at -70 degrees C until analysis. The specimens were tested for the presence of nitric oxide and superoxide dismutase with the Griess and Fridovich methods, respectively. RESULTS: The mean age of the patients in the control group was 45.6 (+/- 13.2) years, and 53.7 (+/- 15.8) years in the pterygium group. We found that nitric oxide and superoxide dismutase levels were significantly higher in the control group (p < 0.05). The level of superoxide dismutase was 747.8 +/- 524.4 units/mg of protein in the normal conjunctiva and 184.2 +/- 163.9 units/mg of protein in the pterygium group. The nitric oxide level was 1.5 +/- 0.9 micromol/mg of protein in the normal conjunctiva and 0.4 +/- 0.4 micromol/mg of protein in pterygium. INTERPRETATION: The low level of nitric oxide may have some role on pterygium development.

The effects of thalidomide in a rat model of surgically-induced endometriosis.

OBJECTIVE: The aim of the study was to analyze the anti-angiogenic role of thalidomide and to assess whether thalidomide had any influence on a rat model of surgically-induced endometriosis. MATERIALS AND METHODS: Endometriosis was induced through surgical induction and homologous transplantation in 16 rats. The rats were randomly separated into two groups as thalidomide (n=8) and control (n=8) groups. Using oral gavage, 100 mg/kg thalidomide 0.5 ml was administered to the first group and saline 0.5 ml to the control group. Histopathologic findings and volume analysis of implants were evaluated after 4 weeks. Vascular endothelial growth factor-A (VEGF-A) and oxidative markers were run from the fluid through peritoneal lavage. RESULTS: The average implant volume decreased significantly in the thalidomide administrated group after treatment (53.3 and 22.9 mm3 respectively, p=0.012). Significant differences observed in the histopathologic scores of the thalidomide group (3 and 1 respectively, p=0.012) were not observed in the control group. Significant decreases were observed in the levels of VEGF-A and myeloperoxidase (MPO) from oxidative markers (p=0.004, p=0.037, respectively). CONCLUSION: Thalidomide provides volumetric and histopathologic recovery in implants particularly because the VEGF inhibition and anti-angiogenic effect, which suggests that it could be effective in the treatment of endometriosis.

The investigation of the effect of Marafl powder (smokeless tobacco) on hematological parameters.

Nicotine is used in different forms including smokeless tobacco. A special kind of smokeless tobacco also known as Maras powder (MP) is widely used in southeastern region, especially Kahramanmaras, Gaziantep and other southeastern cities of Turkey. The aim of this study was to investigate the effect of nicotine on hematological parameters in MP users. Ninety-two MP users from Kahramanmaras and its environs and sixtyeight healthy controls who did not use MP were included in the study. We measured haematological parameters in the blood samples of MP users and controls. Our results showed that while iron and WBC levels were higher in MP users than the controls (p< 0.001), monocyte and platelet counts were lower (p< 0.05 and p< 0.001, respectively). Increased leukocyte counts in MP users may be an indicator of the present inflammatory events in various tissues. So, we assume that MP, because of either high nicotine content or high tobacco-specific nitroso amines levels (TSNA), causes chronic inflammatory changes in various cells, organs and systemic circulation.

Zinc may protect remote ocular injury caused by intestinal ischemia reperfusion in rats.

Remote organ injury represents the oxidative damage, which occurs in various organs away from the tissues exposed to ischemia-reperfusion insult. Hypothesizing that the eye may be susceptible to this type of tissue damage, we investigated the effect of splanchnic ischemia-reperfusion on the chorio-retinal tissue in Sprague-Dawley rats. Four groups, each consisting of 10 male rats, were designed for the study. Ischemia-reperfusion was introduced by clamping superior mesenteric artery for 1 hour, followed by reperfusion for 90 min (IR group). In another group, the same operation was performed except that zinc aspartate (50 mg/kg) was given intra-peritoneally 15 min before the opening of the clamp (Zn-IR group). As control (sham group), the abdomen was opened without any intervention on superior mesenteric artery. The other group was given Zn after opening of abdomen without any intervention on superior mesenteric artery (Zn-sham group). After the choroid and the retina were dissected from the eye, malondialdehyde, superoxide dismutase, catalase and nitric oxide (NO) were measured in the collected eye tissues. We found that the levels of malondialdehyde, an indicator for lipid peroxidation, were higher in the chorio-retinal tissue of IR group compared to Zn-IR, Zn-sham or sham group (p < 0.05). The oxidative stress in Zn-IR group was not higher than that of sham group. Treatment with zinc decreased NO levels but had no noticeable effect on the level of antioxidant enzymes. Our study indicates that remote organ injury is induced in the eye during splanchnic ischemia-reperfusion and that zinc may be beneficial to ameliorate remote ocular injury.

Plasma homocysteine and lipoprotein (a) levels in Turkish patients with metabolic syndrome.

High serum total homocysteine (tHcy) and lipoprotein (a) [Lp(a)] levels are independent risk factors for cardiovascular disease. In this study, we examined the relationship of tHcy and Lp(a) levels with the components of metabolic syndrome. Fifty-one patients diagnosed with metabolic syndrome (median age: 38 [range 25-48] years) and 50 healthy subjects (median age: 35 [26-48] years) were included in the study. We used the National Cholesterol Education Program criteria to define metabolic syndrome. Total tHcy concentrations were measured by using an IMX (Abbott Diagnostics, Abbott Park, IL, USA). Lipoprotein (a) was measured by immunonephelometry using Behring nephrometer method (Behring BN 100, Behring, Germany). Total homocysteine and Lp(a) levels were found to be higher in the metabolic syndrome group than in the control group (tHcy: 24.2 vs 13.4 micromol/l, P < 0.01 and Lp(a): 34.9 vs 15.8 mg/dl, P < 0.01). Vitamin B12 levels were lower in the metabolic syndrome group than in the control group (214 pg/ml vs 247 pg/ml, P < 0.01). In partial correlation, tHcy and Lp(a) concentrations were unrelated to metabolic syndrome or to the components of metabolic syndrome, including fasting serum triglycerides, HDL-cholesterol, fasting glucose, blood pressure, or body mass index. tHcy levels were strongly related only to the vitamin B12 concentration. The risk of cardiovascular disease is higher in patients with metabolic syndrome compared with the normal population. High tHcy and Lp(a) levels should be evaluated in this group of patients in addition to the evaluation of the parameters of metabolic syndrome.

Effects of maternal nicotine exposure during lactation on breast-fed rat pups.

BACKGROUND: Nicotine is known to be associated with adverse effects in infants and children. It is concentrated in breast milk and is absorbed by the infant. The aim of this study was to investigate the effects on breast-fed rat pups of maternal nicotine exposure during lactation. METHODS: In the experimental group (n = 6), nicotine was given to lactating dams (2 mg/kg/day) after delivery and continued for 10 days during lactation. Control animals (n = 4) received saline for the same duration. The suckling rats were weighed and killed on postnatal day 10, and samples were taken from the lung, liver, kidney, spleen and small intestine for histopathological examination. Superoxide dismutase (SOD), catalase (CAT) activities and malondialdehyde (MDA) levels were measured in the liver of the dam and the offspring. RESULTS: Histopathological changes in the liver of the nicotine-exposed group showed portal inflammatory infiltrate, ballooning degeneration of hepatocytes, and focal necrosis in the parenchyma. Thickening of alveolar walls because of interstitial inflammation was noted in the lungs. Histopathological examination of kidney, spleen and small intestine tissue did not reveal any abnormality. In the experimental group, SOD and CAT activities were significantly decreased (p <0.001) but MDA levels were significantly increased (p <0.001) compared with the control group. CONCLUSION: These results indicate that maternal nicotine exposure induces oxidative stress and causes detrimental histopathological changes in the lung and liver of lactating offspring.

Does maternal nicotine exposure during gestation increase the injury severity of small intestine in the newborn rats subjected to experimental necrotizing enterocolitis.

BACKGROUND/PURPOSE: The aim of this study was to evaluate the effects of maternal nicotine exposure during gestation on injury severity of small intestine in the newborn rats subjected to hypoxia-reoxygenation and cold stress. METHODS: A total of 21 Sprague-Dawley pregnant rats were divided into 3 equal groups. The groups were labeled as group 1, control group; group 2, hypoxia-reoxygenation group; and group 3, nicotine-hypoxia-reoxygenation group. The rats of group 3 were exposed to nicotine via subcuticular injection for the last week of gestation (2 mg/kg/d). Newborn rats were collected immediately after birth to prevent suckling of maternal milk (40 rat pups in group 1, 43 rat pups in group 2, and 41 rat pups in group 3). Litters in groups 2 and 3 were stressed twice daily with asphyxia followed by cold (4 degrees C for 10 minutes) stress to induce hypoxic intestinal injury which is relevant to human necrotizing enterocolitis. Breathing 100% CO2 for 10 minutes in a chamber followed by 10-minute 100% O2 breathing was the asphyxia model repeated twice daily. After hypoxia-reoxygenation and cold stress, newborn rats were returned to their mother's cages. This protocol was repeated for the following 2 days, and the rat pups were decapitated on the third day. Using this protocol of asphyxia and cold stress, all of neonatal rats developed clinical and pathological signs of hypoxia-induced intestinal injury. The entire gastrointestinal tract was removed and examined macroscopically. A 2-cm section of distal ileum from each animal was taken for histopathological and biochemical examinations. Histological changes in ileal architecture were scored and graded from 1 to 5. The remaining intestinal tissues of the animals were used for lipid peroxidation analysis. RESULTS: Typical signs of hypoxia-induced intestinal injury were observed in the 2 experimental groups (groups 2 and 3) macroscopically. There were more grades 3 and 4 injuries in group 3 (P < .05). The malondialdehyde levels were elevated in groups 2 and 3 (P < .001). The malondialdehyde levels of the group 3 were also significantly higher than group 2 (P < .01). CONCLUSIONS: Maternal nicotine exposure during gestation results in higher grade histological injury in newborn rats subjected to hypoxia-reoxygenation and cold stress.

The role of urtica dioica (urticaceae) in the prevention of oxidative stress caused by tourniquet application in rats.

Tourniquets are used in extremity surgery and provide a relatively bloodless field, thereby minimizing blood loss and helping identify the vital structures. However, they may cause an ischemia-reperfusion injury with potentially harmful local and systemic consequences. Many therapeutic effects such as diuretic, natriuretic, hypotensive, anti-rheumatic, anti-prostatic, and in-vitro anti-oxidant effects of the Urtica dioica (UD) have been determined. In the present study, we aimed to investigate the potential role of UD plant for prevention of oxidative stress in muscle tissues generated by tourniquet application in rats. Wistar rats were used in this study. The UD extract or 1.15% KCl aqueous solution, in which UD leaf samples were homogenized, was given to each group of eight rats once a day for 5 days through an intraesophageal canule. No treatment was applied to untreated group. Tourniquets were applied to the left posterior limb of rats for 1 or 2 h followed by a reperfusion period of 1 h. After the ischemia and reperfusion, the rats were killed with a high dose of anesthetic drug, and malonyldialdehyde (MDA) levels were measured in their tibialis anterior muscles. Basal MDA levels were obtained from tibialis anterior muscles of 8 control rats, which were not exposed to ischemia. MDA levels were lower in the UD-treated rats than those in untreated and KCl-treated rats after either 1 or 2 h of ischemia and 1 h reperfusion. These results indicate that UD has a potential antioxidant effect on ischemic muscle tissues.

Does long-term ischemia affect the oxidant status during fracture healing?

INTRODUCTION: The influence of transient circulatory arrest on oxidant status during the healing of a tibial fracture was investigated in rats by the use of a hindlimb tourniquet technique. MATERIALS AND METHODS: One of the most reliable indicators for cytological damage is lipid peroxidation, which can be demonstrated by malondialdehyde (MDA) levels. Fifty-eight Wistar rats were used in this study. To determine the basal MDA levels of bone, 10 rats not exposed to ischemia were killed by an overdose of ether. The remaining 48 rats were randomly divided into two groups (control and ischemia). The control and ischemia groups were then randomly divided into 4 groups of 6 rats each. In 48 rats, the left tibia was fractured and fixed intramedullarly. In the ischemic group, complete acute transient ischemia for 4.5 h was imposed after the fracture. In the control group, no other intervention except the fracture was done. Rats from the control and ischemic groups were killed on days 3, 7, 14, and 28, and MDA levels were determined in tibial bone and callus tissue. The MDA levels of the control and ischemic groups were compared with basal MDA levels in the bone of 10 rats. RESULTS: There was an apparent difference between the basal and control group MDA levels on days 3 and 7 (p < 0.01), between the basal and ischemic group MDA levels on days 3, 7, and 14 (p < 0.01). In addition, the ischemic group showed a statistically significant increase in MDA levels on days 3, 7 and 14 compared with the control group (p < 0.01). CONCLUSION: We conclude that complete acute transient ischemia affects the oxidant status during fracture healing. This effect especially occurs during the ischemic period, inflammation, and callus formation of fracture healing.

Relationship between oxidative stress in cord blood and route of delivery.

OBJECTIVE: To investigate the effect of route of delivery on the antioxidant system of newborns. METHODS: We used the cord blood taken during labor of 56 vaginal deliveries and 50 elective cesarean sections. The specimens were analyzed for the plasma malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glucose 6-phosphate dehydrogenase (G6PD) activity and leukocyte count. RESULTS: SOD and CAT activities were significantly higher in the elective cesarean group than the vaginal delivery group (p < 0.01 and p < 0.005, respectively), but G6PD activity was similar between the groups. Plasma MDA level was lower in the cesarean group compared to the vaginal delivery group (p < 0.001). Leukocyte count was higher in the vaginal delivery group (p < 0.005). CONCLUSION: Our findings suggest that the route of delivery has an effect on oxidative stress in newborns exposed to oxidative stress during delivery. It can be suggested that the antioxidant system works more efficiently to overcome oxidative stress in newborns delivered via cesarean section.

Low-volume tumescent liposuction does not change plasma malondialdehyde levels: a preliminary study.

To evaluate the effects of low-volume tumescent liposuction on plasma malondialdehyde (MDA) levels, a controlled clinical study was conducted. Venous blood samples were taken just before and 10 min after the completion of tumescent suction-lipectomy for 15 patients. The means +/- standard deviations for patients' data were as follows: of patients' age (29.81 +/- 13.2 years) aspirate volume (1,250 +/- 450 ml), and body mass index (24.5 +/- 1.2). There was no significant difference between the preoperative and postoperative plasma malondialdehyde levels (p > 0.05). The results show no evidence of increased lipoperoxidation in the systemic circulation during low-volume tumescent liposuction.