Osman Caglayan

Increased tumor necrosis factor alpha (TNF‐α) and interleukin 1 alpha (IL1‐α) levels in the lesional skin of patients with nonsegmental vitiligo

Increased tumor necrosis factor alpha (TNF-α) and interleukin 1 alpha (IL1-α) levels in the lesional skin of patients with nonsegmental vitiligo Dear Sir, Depigmentation in vitiligo is caused by melanocyte destruction. There are different hypotheses including neural, selfdestruction, and immune hypotheses to explain the pathogenesis of vitiligo There is growing evidence that cytokines are important in the depigmentation process of vitiligo. Granulocytemacrophage colony-stimulating factor (GM-CSF), endothelins, b-FGF are the mitogens for melanocytes whereas TNFα, IL1α, IL-6, TGFβ are the potent inhibitors of melanocyte growth. IL1α is remembered as one of the cytokines of inflammation, and is also known as a B-cell activating factor. It produces similar biological effects with TNF α and is produced in most inflammatory and immunological diseases. b-FGF, produced by fibroblasts and epidermal keratinocytes, is a polypeptide and is capable of promoting angiogenesis and mitogenesis through autocrine and paracrine mechanisms. The role of peripheral blood and lesional cytokine expression in patients with vitiligo has not been clarified yet. Herein we wanted to investigate the levels of cytokines in the skin and serum of vitiligo patients and to compare them with the levels in healthy controls. Six female and 18 male vitiligo patients aged (32 years ± 17 SD) were enrolled in the study after giving informed consent. Thirteen (54.2%) had symmetrical generalized, seven (29.2%) had acrofacial, and four (16.7%) had focal vitiligo. Six (25%) had progressive and 18 (75%) had stabile vitiligo (no new depigmentation had been observed for more than 3 months). Fourteen ageand sex-matched healthy volunteers consisted the control group. Samples of peripheral blood from patients and healthy volunteers were collected by venipuncture. Serum was separated and stored at −70 °C before use. Four-millimeter punch biopsies were taken in all patients from lesional skin and nonlesional skin (at least 3 cm far from lesional biopsy) and from healthy volunteers. The biopsies were frozen in liquid nitrogen (−196 °C) and preserved at −70 °C. Levels of IL1α, TNFα, and b-FGF were measured with specific ELISA kits (Biosource International, Camarillo, California, USA) according to the manufacturer’s instructions. Skin cytokines were measured as described by Lowry et al. Level of the cytokines were expressed as μmol/mg protein. Statistical analysis was carried out using spss 10.0. Differences in values of cytokine amount between lesional vs. healthy volunteers, nonlesional vs. healthy controls, serum of patients vs. serum of healthy volunteers were carried out using Mann– Whitney U-test. The parameters of lesional vs. nonlesional skin were analyzed by the Wilcoxon-signed rank test. P-value of less than 0.05 was considered to be statistically significant. The mean (± SD) values of IL1α, TNFα, b-FGF in lesional, nonlesional, healthy skin, and the serum from healthy controls and the study group are given in Table 1. The expression of IL1α and TNFα was significantly higher in lesional skin than in nonlesional skin in patients with vitiligo (P = 0.007 and P = 0.002), respectively. The exact mechanism how cytokines effect pigmentation is not fully understood. The different hypotheses are: (a) TNFα induces IL1α promoting B-cell differentiation and immunoglobulin production. (b) Cytokines such as IFNγ, TNFα, TNFβ, IL1α, and IL-6 can induce cell surface ICAM-1 on melanocytes which is necessary for leukocyte–melanocyte attachment. ICAM-1 may also induce B-cell activation, increasing autoantibody production and may cause melanocyte damage in vitiligo. (c) TNFα and IL1α also have the capacity to induce apoptosis in many cell types. (d) Melanogenesis is also inhibited by TNFα through an inhibitory effect on tyrosinase and tyrosinase related protein. Moretti et al. found increased levels of IL-6 and TNFα in the epidermis of lesional skin compared with the healthy controls. Swope et al. investigated the role of epidermal cytokines in pigmentation and found that IL1α, TNFα, and IL-6 elicited a dose-dependent decrease in the activity of the enzyme tyrosinase of cultured normal human melanocytes and also inhibited melanocyte proliferation. Ozdemir et al. demonstrated increased skin blister fluid b-FGF levels and serum levels in vitiligo patients compared with healthy controls and proposed that b-FGF plays a role in the pathogenesis of vitiligo. However,

Oxidative Stress Markers in Young Patients with Polycystic Ovary Syndrome, The Relationship between Insulin Resistances

OBJECTIVE Polycystic ovary syndrome is a syndrome of ovarian dysfunction. Oxidative stress, inflammation and endothelial cell activation are thought to play concomitant roles in the pathogenesis of the above diseases particularly in the development of atherosclerotic lesions. RESEARCH DESIGN AND METHODS We studied 58 polycystic ovary syndrome patients and age-matched 25 healthy controls consisting of women that have regular, ovulatory cycles and normal androgen levels. Homeostasis Model Assessment-Insulin Resistance for this study was taken as 1.75 that is the upper level of confidence interval of %95 of the mean of the healthy group. PCOS patients were divided into two groups as for below the cut-off level (<1.75) and above the cut-off level (> or =1.75). hs-CRP, fibrinogen, malondialdehyde, nitric oxide and disulfide level results were compared both in PCOS and control groups. RESULTS In this study, sensitive CRP was found to be statical significantly higher in polycystic ovary syndrome groups whose Homeostasis Model Assessment-Insulin Resistance were > or =1.75 and <1.75 when compared to the control group. But, no significantly correlation was determined between malondialdehyde, nitric oxide and disulfide levels and CRP elevation. CONCLUSIONS In our study, because those participants were young and non- obese patients with PCOS, malondialdehyde, nitric oxide and disulfide levels and Carotid Artery Intima-Media Thickness measurements as a pre-indicator of cardiovascular disease were not found to be different from those of the controls.

Plasma D -Lactate Levels in Diagnosis of Appendicitis

We investigated the possible use of D -lactate as a predictor in the diagnosis of appendicitis. C-reactive protein level (CRP) and leukocyte counts were also evaluated. Venous blood D -lactate, CRP, and leukocyte counts were measured preoperatively in 53 patients undergoing surgery for appendicitis, as well as in 20 healthy subjects. Levels of all three parameters in the surgical patients were significantly higher than in the control group ( p < .05). Previous studies have shown that venous D -lactate is more specific to the intestine than CPR or leukocyte count. Based on our data, venous D -lactate, which had the lowest false-negative rate among these laboratory parameters, may be a useful diagnostic marker for appendicitis. None of these parameters were helpful in identifying the type of the appendicitis.

Intestinal ischemia-reperfusion and plasma enzyme levels.

Abstract Determination of blood levels of intracellular enzymes is an appropriate method to evaluate tissue and organ damage. To show systemic tissue damage resulting from intestinal ischemia-reperfusion, New Zealand rabbits underwent 60 min intestinal ischemia and 60 min reperfusion. Plasma samples were obtained before and at 55, 70, and 120 min after operation and enzyme levels were determined. Plasma aspartate aminotransferase (AST) showed a significant increase during reperfusion while lactate dehydrogenase (LDH) and creatine kinase (CK) levels were significantly increased at the end of ischemia and continued to be so throughout reperfusion. It is difficult to claim that enzymes arise from the intestine, but an increase of CK, LDH, and later of AST without any increase in alanine aminotransferase levels during ischemia suggests that their primary source is the injured intestine. Increased levels of plasma enzymes do not provide exact information about the location, but do reveal the presence of an injury.

Effect of vitamin E and C supplementation combined with oral antidiabetic therapy on the endothelial dysfunction in the neonatally streptozotocin injected diabetic rat

This study investigates the contribution of vitamin supplementation to the efficacy of oral antidiabetic therapy on the reversal of endothelial dysfunction in a model of type‐2 diabetes in rat.

The relationship of the interleukin-6 -174 G>C gene polymorphism with oxidative stress markers in Turkish polycystic ovary syndrome patients.

Objective: Interleukin-6 (IL-6) is a key pro-inflammatory and immune-modulatory cytokine of relevance for cardiovascular (CD) diseases. Cardiovascular risk factors that have been reported include oxydative stress markers [nitric oxide (NO), malondialdehyde (MDA), disulphite (SH)]. We aimed to evaluate the relation between the IL-6 G/C gene polymorphism and oxidative stress markers in polycystic ovary syndrome (PCOS) patients. Design and patients: We studied 85 PCOS patients and 115 healthy controls. PCOS was defined by the Rotterdam PCOS consensus criteria. Results: The genotype IL-6 distribution did differ between the control group (CC 9.6%, GC 63.4%, GG 27.0%) and the PCOS patients (CC 4.7%, GC 29.4%, GG 65.9%) (p<0.001). The frequency of the polymorphic G allele was also not similar for the group with PCOS as for the control group with 80.6% and 58.7%, respectively (p<0.001). No statistically significant difference was determined for MDA and NO levels in PCOS patients and control group (p>0.05). Only SH levels were found to be high in favor of patient group (p<0.05). No statistically significant difference was determined between IL-6 G/C gene polymorphism and oxidative stress markers in PCOS patients and in the control group. Conclusion: Gene polymorphism of IL-6 −174 G>C is a risk factor for PCOS in Turkish patients. IL-6 gene polymorphisms are not related to NO, MDA, and SH levels in PCOS. Our negative results in risks factors of CV disorders can probably be explained by the fact that metabolic parameters and endothelial systems of patients may not yet be affected in this short period of time.

Similar effects of general and spinal anaesthesia on perioperative stress response in patients undergoing haemorrhoidectomy.

Surgery induces release of neuroendocrine hormones (cortisol), cytokines (interleukin-6: IL-6, tumour necrosis factor-α: TNF-α), acute phase proteins (C-reactive protein: CRP, leptin). We studied the effects of general and spinal anaesthesia on stress response to haemorrhoidectomy. Patients were assigned to general and spinal anaesthesia groups (n = 7). Blood samples were drawn before induction and 24 hours after surgery. Perioperative levels of IL-6, TNF-α, CRP, cortisol, and leptin were comparable among the groups. Twenty four hours after surgery, TNF-α and cortisol did not change; IL-6 and CRP increased significantly in all patients. Significant increase in leptin levels was found in patients undergoing spinal anaesthesia. Except for the increase in leptin levels, there was no significant difference related to the effects of general and spinal anaesthesia.

Effect of I-deprenyl and gliclazide on oxidant stress/antioxidant status and dna damage in a diabetic rat model.

Background: This study investigates the possible effect of monoamine oxidase inhibitor (MAOI), selegyline (l-deprenyl), in combination with oral antidiabetic-gliclazide (OAD), in preventing oxidative stress in streptozotocin-induced diabetes model in male Swiss Albino rats by measuring oxidant stress/DNA damage and antioxidant levels. Methods: Diabetic rats were divided into four groups (n = 10) as [1] diabetic untreated (DM), [2] deprenyl treated (DM + D), [3] gliclazide treated (DM + O), and [4] gliclazide and deprenyl treated (DM + O + D). Controls were divided into two groups (n = 8) [1] untreated (C), and [2] deprenyl treated (C + D). Gliclazide 5 mg/kg and/or MAOI 0.25 mg/kg daily were given orally by gavage for 4 weeks. At the end of the 12th week, catalase and superoxide dismutase (SOD) levels in erythrocyte lysates (EL); total antioxidant status (TAS), 8-hydroxy-deoxyguanosine (8-OHdG), malondialdehyde (MDA), and vitamin A and E levels in plasma, MDA, and MAO in liver homogenates were determined. Results: Diabetic rats showed a decrease in EL-SOD, plasma TAS, and vitamin E, and an increase in plasma 8-OHdG, plasma, and liver MDA levels (p < 0.05). Gliclazide and/or deprenyl decreased 8OHdG levels and increased antioxidant levels and survival when compared with untreated diabetic rats (p < 0.05). The lowest 8-OHdG levels were determined in the DM + O + D group. Conclusions: The combined treatment of deprenyl and gliclazide may contribute to the control of the physiopathological mechanisms underlying both the process of aging and type 2 diabetes by reducing oxidant stress and DNA damage, improving antioxidant status, and increasing survival, and may have implications for further clinical studies.

Effects of ovariectomy and ascorbic acid supplement on oxidative stress parameters and bone mineral density in rats.

Objectives The aim of this study is to investigate the effects of ovariectomy on bone mineral density (BMD) and oxidative state in rats, and the alterations in these effects that vitamin C supplementation may produce. Materials and methods Twenty female Wistar albino rats were randomly divided into three groups: control (C, n=6); ovariectomy (O, n=7); and ovariectomy + vitamin C supplement (OV, n=7). Oxidative stress (OS) was assessed 100 days postovariectomy by measuring the activity of several enzymes, including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase, as well as the concentrations of malondialdehyde (MDA), nitric oxide (NO), and total sulfhydryl groups in plasma and bone homogenates. Results A significant decrease in BMD was observed in O group compared with C group (p = 0.015), and a significant increase was observed in OV compared with O group (p=0.003). When groups were compared with respect to parameters of OS, MDA and NO levels in bone tissue were significantly higher in O than in C (p=0.032, p=0.022) and were significantly lower in OV than in O (p=0.025, p=0.018). SOD activity was significantly higher in O than in C (p=0.032). In plasma, MDA activity was significantly higher in O than in C (p = 0.022) and NO level was significantly higher in O than in C and OV (p=0.017, p=0.018). Conclusions Our results suggest that ovariectomy may produce osteoporosis and OS in females, and vitamin C supplementation may provide alterations regarding improvement in OS and BMD values. We assume that studies including more subjects are needed to make a decisive conclusion about OS–BMD relation.

Periodontal health in children exposed to passive smoking

AIM To determine (1) the cotinine levels of saliva, urine and gingival crevicular fluid (GCF) of children in families with and without smoking members and (2) a possible association between the periodontal health of the children and exposure to passive smoking. MATERIAL AND METHODS The study population comprised of 109 children in the age range 6-12 years. Children were classified as exposed to passive tobacco smoking (PTS-exposed, n=51) and as unexposed controls (PTS-unexposed, n=58). Plaque index, gingival index, bleeding on probing, probing depth and clinical attachment level (CAL) were recorded. GCF, saliva and urine samples were also collected. The levels of cotinine in these fluids were determined by enzyme-linked immunosorbent assay. RESULTS The mean salivary cotinine concentration was significantly increased in PTS-exposed children compared with PTS-unexposed children (p<0.05). Further, in a dose-dependent way, the mean salivary concentration was significantly higher in children whose father or mother was a smoker (p<0.05) as compared, respectively, with children whose fathers and mothers were non-smokers. The mean CAL was significantly less in PTS-exposed children compared with non-PTS-exposed children (0.09 mm; p<0.05) and also in children whose father was a smoker (p<0.05), but not in children whose mother was a smoker as compared with non-smoker fathers and mothers, respectively. The GCF cotinine levels were below the detection limits with the assay method that was used. CONCLUSIONS We have observed that children who are exposed to passive smoking have elevated cotinine levels in their saliva concomitant with a lowered CAL.