Estrogen as primary factor in vaginal healing in rats.

OBJECTIVE: Although the effect of estrogens on wound healing is already known, its complex mechanism is not fully understood in literature. The aim of this study is to investigate the effect of estrogen on vaginal healing after surgical intervention performed in the age group with low estrogen level and in an adult group with high estrogen level. MATERIALS AND METHODS: Seven young and seven adult female Wistar Albino rats were procured. For control group, one animal was chosen each from the young (Group I) and adult groups (Group II), and their vaginal tissue was removed. An incision was made to the posterior vaginal wall under anesthesia and sutured with 5-0 polyglactin in all the rats. On the seventh postoperative day, the posterior vaginal wall was excised. A semi-quantitative method was used to evaluate the histological processes and structures during wound healing. RESULTS: Although there were no evident differences in the evaluation of histological scoring system, the presence and distribution of new vascularization and fibroblasts showed that vaginal mucosal healing was more intense in adult rats. CONCLUSIONS: The effect of estrogens on vaginal mucosal healing has been discussed in several experimental studies and literature information has been presented; it has been concluded that it would be beneficial to consider the positive effect of vaginoplasty procedures.

Oxysterol species: reliable markers of oxidative stress in diabetes mellitus.

PURPOSE: To assess the plasma oxysterol species 7-ketocholesterol (7-Kchol) and cholestane-3ß,5α,6ß-triol (chol-triol) as biomarkers of oxidative stress in type 1 and type 2 diabetes mellitus (DM). METHODS: In total, 26 type 1 and 80 type 2 diabetes patients, along with 205 age- and gender-matched healthy controls, were included in this study. Oxysterols were quantified by liquid chromatography coupled with tandem mass spectrometry and N,N-dimethylglycine derivatization. Correlations between oxysterols and clinical/biochemical characteristics of the diabetes patients, and factors affecting 7-Kchol and chol-triol, were also determined. RESULTS: Plasma 7-Kchol and chol-triol levels were significantly higher in type 1 and type 2 diabetes patients compared to healthy controls (P < 0.001). Significant positive correlations were observed between oxysterol levels and levels of glycated hemoglobin (HbA1c), glucose, serum total cholesterol, low-density lipoprotein, very-low-density lipoprotein, and triglycerides, as well as the number of coronary risk factors. Statins, oral hypoglycemic agents, and antihypertensive agents reduced the levels of oxysterols in type 2 diabetes patients. Statin use, HbA1c levels, and the number of coronary risk factors accounted for 98.8% of the changes in 7-Kchol levels, and total cholesterol, smoking status, and the number of coronary risk factors accounted for 77.3% of the changes in chol-triol levels in type 2 diabetes patients. CONCLUSIONS: Plasma oxysterol levels in DM, and particularly type 2 DM, may yield complementary information regarding oxidative stress for the clinical follow-up of diabetes patients, especially those with coronary risk factors.

Neutrophil volume distribution width as a new marker in detecting inflammatory bowel disease activation.

INTRODUCTION: We sought to investigate the value of neutrophil volume distribution width in detecting inflammatory bowel disease activation. METHODS: Patients with infection and accompanying inflammatory disease were excluded. All the patients were diagnosed and classified according to Porto criteria and Paris classification, respectively. Physician global assessment, pediatric Crohn's disease and pediatric ulcerative colitis activity indexes and fecal calprotectin were used to define disease activation. RESULTS: A total of 34 pediatric patients with Inflammatory bowel diseases (IBD) and 29 controls were enrolled in the study. Neutrophil volume distribution width (NVDW) was significantly higher in patients with IBD compared to healthy controls (P < 0.001). An increased NVDW level was observed in IBD patients in activation (22.42 ± 2.13) compared to those in remission (19.22 ± 1.63) (P < 0.001). There was no statistically significant difference between IBD patients in remission and healthy controls. The best cutoff of NVDW for prediction of disease activation in Crohn's disease and ulcerative colitis in this series was 20.39 with a sensitivity of 90.9% and a specificity of 75% (AUC: 0.852 CI: 0.698-1.000 P < 0.001) and 19.74 with a sensitivity of 92.9% and a specificity of 90.9% (AUC: 0.961, CI: 0.889-1.000, P < 0.001), respectively. CONCLUSIONS: As a quantitative, objective, and sensitive parameter, we believe that the NVDW has a potential to be an additional test detecting disease activation in IBD.

Diagnostic performances of CA125, HE4, and ROMA index in ovarian cancer.

PURPOSE OF INVESTIGATION: HE4 (human epididymis protein 4) is suggested to be used as a potential new biomarker to identify ovarian malignancies from benign adnexal masses. The aim of this study was to evaluate HE4, in comparison with CA125 and Risk of Ovarian Malignancy Algorithm (ROMA) index in benign gynecological diseases and ovarian cancer, and additionally to determine the reference range for HE4 in healthy Turkish women. MATERIALS AND METHODS: CA125 and HE4 serum levels were determined in 96 patients with benign gynecological diseases, 47 patients with ovarian cancer and 106 healthy women using a specific analyzer. CA125 and HE4 cut-offs were 35 U/ml and 70 pmol/L, respectively. RESULTS: HE4 had significantly higher concentrations in ovarian cancer than benign gynecologic disorders (p < 0.005). Tumor marker sensitivity in ovarian cancer was 78% for HE4, 63% for CA125, and 88% for ROMA index at 95% specificity. A significantly higher area under the Receiver operator characteristic (ROC) curve was obtained with HE4 and ROMA index than CA125 in the differential diagnosis of benign gynecological diseases versus ovarian cancer (0.929, 0.955, and 0.781, respectively). Reference limits for serum HE4 in healthy Turkish women was determined as 28.9-62.4 pmol/L for pre-menopausal and 23.7-152.4 pmol/L for postmenopausal women. CONCLUSIONS: In the diagnosis of ovarian cancer, HE4 had higher sensitivity, as a single tumor marker. The sensitivity of HE4 and ROMA index in postmenopausal women was higher than premenopausal women for detecting ovarian cancer.

Does tamoxifen citrate prevent pulmonary fibrosis due to silica inhalation?

BACKGROUND: As shown in several studies, besides being used in breast cancer, tamoxifen is also known for its antifibrotic effects via reducing the serum TGF-beta levels. We investigated the possible preventive effect of tamoxifen in rats exposed to silica particles depending on the antifibrotic effect. MATERIALS AND METHODS: A total of 102 adult female Wistar Albino rats were divided into five groups. First two groups (control and tmx) were free of silica and the last three groups (slc, tmx1 and tmx 10) were exposed to crystalline silica. The rats in tmx, tmx1 and tmx10 groups received 10 mg/kg, 1 mg/kg and 10 mg/kg of body weight tamoxifen, respectively. On day 84, all rats were sacrified and tissue samples were obtained together with blood samples. The differences in serum TGF-ß levels, histological grades of fibrosis and inflammation in the lung and liver tissues together with addional biochemical markers were calculated between the groups. RESULTS: Silicosis occurred in slc, tmx1 and tmx10 groups in 100%, 91.7% and 52.1%, respectively. Liver fibrosis did not occur. The highest mean lung fibrosis scores were obtained in slc group while the scores were lower in tmx1 group and the lowest in tmx10 within silica-exposed rats. Nevertheless, the inflammation scores were higher in tamoxifen-administered rats in a dose-dependent pattern. CONCLUSION: Silica inhalation did not result in liver fibrosis. Tamoxifen is found to prevent lung fibrosis and reduce serum TGFß-1 levels while increasing lung inflammation (Tab. 3, Fig. 3, Ref. 27).

The effect of taurine on mesenteric blood flow and organ injury in sepsis.

Endotoxin decreases mesenteric blood flow and inflicts organ injury via free radicals. We investigated whether taurine, an endogenous antioxidant and vasodilator, could attenuate the deleterious effects of endotoxin in a mouse model of sepsis. Swiss albino mice were allocated into four groups and treated either with taurine (150 mg/kg, i.p. at 0(th), 8(th), 16(th) h) or its solvent sterile saline (NaCl 0.9%, w/v) while E. coli endotoxin (20 mg/kg, i.p.) or its solvent saline were also given at 8(th) h. At 24(th) h the animals were anaesthetized and the mesenteric blood flow was measured by using perivascular ultrasonic Doppler-flowmeter. The animals were then exsanguinated, the spleen, liver, and kidneys were isolated for histopathological examination. Thiobarbituric acid-reacting substances (TBARS), glutathione, and myeloperoxidase activity were determined in the liver samples. Endotoxin significantly decreased the mesenteric blood flow and glutathione levels in liver while TBARS and myeloperoxidase activity were increased. However, taurine did not block the deleterious effects of endotoxin nor it did attenuate the histopathological injury. Therefore, we concluded that endotoxin-induced organ injury via free radicals is resistant to blockade by taurine.

Blockade of sodium channels by phenytoin protects ultrastructure and attenuates lipid peroxidation in experimental spinal cord injury.

Spinal cord injury (SCI) involves a series of pathological events. Abnormal sodium influx has been implicated as one of the key events in the pathophysiology of the SCI. Pharmacological blockade of sodium channels can reduce secondary injury and increase recovery from trauma. The aim of the present study was to show the neuroprotective effect of phenytoin, a sodium channel blocker, after experimental SCI. Control and laminectomy-only groups were not injured. 50 g-cm weight drop injury was produced in the trauma group. In the treatment groups, methylprednisolone (30 mg/kg) and phenytoin (1 mg/kg, 10 mg/kg, or 30 mg/kg) were given intraperitoneally immediately after injury. Malondialdehyde (MDA) levels in the spinal cord samples were examined for lipid peroxidation. Spinal cord ultrastructure was evaluated and grading system was used for quantitative evaluation. Trauma increased tissue MDA levels. Treatment with methylprednisolone and phenytoin decreased MDA levels compared to trauma in all doses. Significant ultrastructural neuroprotection was observed with 30 mg/kg of phenytoin treatment according to general neural score. This ultrastructural neuroprotection of phenytoin was not different from methylprednisolone. Phenytoin appears to protect spinal cord against injury by decreasing lipid peroxidation and lessening neuronal damage associated with SCI in rats.

Ligand-induced expression of peroxisome proliferator-activated receptor alpha and activation of fatty acid oxidation enzymes in fatty liver.

BACKGROUND: Peroxisome proliferator-activated receptor alpha (PPARalpha) regulates lipid metabolism upon activation by ligands. Peroxisome proliferator-activated receptor alpha may play a role in the pathogenesis of fatty liver disease. The aim of this study was to assess the PPARalpha expression pattern and mitochondrial/peroxisomal enzyme activities in response to high fat diet (HFD) and clofibrate, a well known PPARalpha ligand. MATERIALS AND METHODS: Four groups of Wistar-Albino rats were included: (1) rats fed a control diet (CD) for 6 weeks, (2) rats fed CD (6 weeks) plus clofibrate (last 2 weeks), (3) rats fed HFD for 6 weeks, and (4) rats fed HFD (6 weeks) plus clofibrate (last 2 weeks). Peroxisome proliferator-activated receptor alpha expression was evaluated by immunohistochemistry. Fatty acid beta-oxidation (peroxisomal-acyl-CoA-oxidase and mitochondrial-acyl-CoA-dehydrogenase) and catalase enzyme activities, and malondialdehyde and glutathion levels were measured spectrophotometrically in liver tissues. RESULTS: All animals were fed HFD but only 2/12 animals were fed HFD plus clofibrate-developed fatty liver. Both HFD and clofibrate induced PPARalpha expression, clofibrate induction being more prominent than HFD. Clofibrate plus HFD did not further increase PPARalpha expression. Activities of peroxisomal-acyl-CoA-oxidase and mitochondrial-acyl-CoA-dehydrogenase enzymes were not induced by HFD alone. Clofibrate increased the activity of these enzymes in both CD- and HFD-fed animals. However, an increase of acyl-CoA-oxidase activity was blunted in rats fed HFD. Catalase activity and malondialdehyde levels were increased but glutathion levels were unchanged in rats fed HFD plus clofibrate. CONCLUSIONS: Clofibrate was a more potent inducer of PPARalpha expression than HFD in our rat fatty liver model. The finding of blunted peroxisomal enzyme response to clofibrate in fatty livers suggests that alterations in postreceptor events may exist and further contribute to liver steatosis. Clofibrate seems to stabilize glutathion content and this might contribute to the prevention of liver steatosis.

Elevated levels of nitrate in rheumatoid arthritis.

Nitric oxide (NO) is a free radical that plays important roles in many physiological and pathological processes. Evidence suggests that NO participates in the pathogenesis of inflammatory reactions in many autoimmune and inflammatory diseases such as rheumatoid arthritis (RA). The purpose of this study was to evaluate serum concentrations of NO in patients with RA and to determine whether they correlate with clinical and laboratory parameters of RA disease activity. Twenty-seven RA patients were recruited for the study and compared with 20 healthy subjects. Serum NO concentrations were measured indirectly in terms of nitrate using colorimetric assay. Disease activity was determined by laboratory and clinical findings. Mean serum concentrations of nitrate were significantly higher than those of healthy controls (P < 0.05). Among the disease activity parameters, C-reactive protein, number of swollen and tender joints, Ritchie articular index, and disease activity scores correlated significantly with serum NO levels. Our results suggest that these levels can serve as a reliable parameter of disease activity in patients with RA. Further knowledge about the precise role of NO may lead to better understanding of the pathogenesis of RA. Furthermore, modulation of NO synthesis may represent a new approach to the treatment of inflammatory and autoimmune conditions.

In vitro and in vivo inhibition of myeloperoxidase with 5-fluorouracil.

OBJECTIVE: Myeloperoxidase (MPO) exists in neutrophils and has an important bactericidal role. During phagocytosis, MPO catalyzes a peroxidative reaction using chloride ion and hydrogen peroxide (H2O2) as substrate. The aim of the present study was to investigate whether 5-fluorouracil (5-FU), a chemotherapeutic agent, has a direct inhibitory effect on MPO and to evaluate some properties of this inhibition. METHODS: The inhibitory effect of 5-FU on MPO was studied in rat tissue, human leukocytes, and leukocytes from cancer patients under 5-FU therapy. MPO was solubilized in a detergent-containing buffer. MPO activity was measured spectrophotometrically through the oxidation of a synthetic substrate tetramethyl benzidine in the presence of H2O2. RESULTS: 5-FU inhibited tissue-associated MPO activity in a dose-dependent but not time-dependent manner with an IC50 value of 0.6 mg/ml. 5-FU also inhibited MPO activity in isolated human leukocytes in a dose-dependent manner, and the IC50 value was 0.75 mg/ml. Using this 5-FU concentration, the inhibitory effect was monitored at different substrate concentrations. Leukocyte MPO activities of patients receiving 5-FU therapy were compared before treatment and after the first, second, and third administration cycles. 5-FU treatment resulted in a significant decrease in leukocyte MPO activity, and repeated 5-FU treatment caused additional decrease. CONCLUSION: Our data showed that 5-FU directly inhibited the MPO activity of human leukocytes in vitro and in vivo. We concluded that, the patients treated with 5-FU should be intensively followed for the risk of infections.

Type 1 sirenomelia in one of male twins, with imperforate anus in the other male twin.

The etiology of sirenomelia sequence is still obscure. The role of maternal diabetes and a vascular steal phenomenon have been discussed [Gürakan et al. (1996) Turk J Pediatr 38:393-397]. Discordant monozygotic twin sirenomelia has been commonly reported but only rarely in dizygotic twins. The family of the presented twins had a high risk of diabetes mellitus. One of the twins has type 1 sirenomelia and the other had only an imperforate anus.

Ischaemia-reperfusion injury of the peripheral nerve: An experimental study.

Although the neuropathology of ischaemic fibre degeneration is relatively well known, its pathogenesis is poorly understood. One of the presumed mechanisms is oxidative stress, causing the breakdown of the blood-nerve barrier (BNB) and ending in lipid peroxidation. We evaluated the effect of ischaemia and reperfusion on the sciatic-tibial nerve of the rat and investigated the biochemical, pathological, and functional evidence of BNB disruption and lipid peroxidation. The distal portion and trifurcation of the sciatic nerve were rendered ischaemic by clamping the femoral vessels for 3 h and followed by varying durations of reperfusion. Reperfusion resulted in an increase in lipid peroxidation beginning from the first hour and increasing until the seventh day, followed by a gradual decline over the following weeks. Nerve oedema and ischaemic fibre degeneration (IFD) consistently became more severe and prominent with reperfusion, indicating that oxidative stress damages the BNB and causes IFD. Results of functional testing by the sciatic function index correlated with other parameters as walking track analysis results got worse as reperfusion periods increased. Impairment of walking patterns was more striking after the first day and continued up to the third week. These data indicate that severe ischaemia of the peripheral nerve results in reperfusion injury, functional impairment, and disruption of the BNB. Microvascular events, which may occur during reperfusion, may be important in amplifying the nerve fibre degeneration that initiated during ischaemia.

Effect of mexiletine on lipid peroxidation and early ultrastructural findings in experimental spinal cord injury.

OBJECT: The purpose of this study was to investigate the effect of mexiletine on lipid peroxidation and on ultrastructural findings after induced spinal cord injury (SCI). The authors also compared the activity of mexiletine to that of the well-known antioxidant, methylprednisolone sodium succinate (MPSS). METHODS: Wistar rats were divided into seven groups, (Groups 1-7). Those in Groups 1 and 2 were control animals that underwent laminectomy only, after which nontraumatized spinal cord samples were obtained immediately (Group 1) and 2 hours postsurgery (Group 2). Spinal cord injury was induced in all other groups, and cord samples were obtained at 2 hours postsurgery. The rats in Group 3 underwent SCI alone; those in Group 4 received 30 mg/kg of MPSS intraperitoneally immediately after trauma was induced; and those in Groups 5, 6, and 7 received 1, 10, and 50 mg/kg of mexiletine, respectively, by intraperitoneal injection immediately after trauma was induced. Compared with the levels in control animals, lipid peroxidation was significantly elevated in rats in Groups 3 and 5, but there were no statistical differences among those in Groups 1, 2, 4, 6 and 7 in this regard. Compared with the findings in rats in Group 3, ultrastructural damage post-SCI was minor in rats in Groups 4 and 5, and there was even less damage evident in rats in Group 7. CONCLUSIONS: Analysis of these findings showed that administration of 50 mg/kg mexiletine significantly decreased the level of lipid peroxidation and protected spinal cord ultrastructure following SCI.