Assessment of COL1A1 and MMP9 expression in patients with dermatochalasis.

PURPOSE: Dermatochalasis is a clinical condition characterized by loss of elasticity of eyelid skin and soft tissue, which typically affects the elderly population. The aim of this study is to investigate the mRNA expression levels of collagen type 1 alpha 1 (COL1A1) and matrix metalloproteinase-9 (MMP9) genes in dermatochalasis tissue. METHODS: The study group consisted of 15 patients who underwent upper eyelid blepharoplasty and were above 40 years old. The patients in our control group were divided into two subgroups according to their ages. Fourteen patients who were under 40 years old and had anterior blepharoptosis surgery for blepharoptosis were designed as the young control group. Sixteen patients who were older than 40 years old and had anterior blepharoptosis surgery for blepharoptosis were designed as the old control group. The patients in the dermatochalasis group were also evaluated according to their smoking status. Surgical tissue specimens were analyzed for COL1A1 and MMP9 mRNA gene expression levels by using real-time polymerase chain reaction. RESULTS: COL1A1 and MMP9 mRNA gene expression levels were not statistically different between the groups (p = 0.247; p = 0.052, respectively). When compared in means of the smoking habit, smokers in the dermatochalasis group exhibited higher COL1A1 mRNA expression levels when compared to nonsmokers (p = 0.008). MMP9 gene expression levels of smokers exhibited almost statistically higher levels but at the limit when compared to nonsmokers (p = 0.05). CONCLUSIONS: This study represents a preliminary study to detect the tissue changes at a molecular level in dermatochalasis, which is known to be related to connective tissue pathology. Collagen and MMPs are essential components of the extracellular matrix, and smoking might affect their gene expression. Further prospective studies on these regulatory genes and encoded protein levels with a larger group of patients may provide particular contribution to explaining the pathophysiology of dermatochalasis.

The Effect of Sildenafil on Selenite-Induced Cataract in Rats.

PURPOSE: To investigate the effect of sildenafil on an experimental sodium selenite-induced cataract model in rats. MATERIALS AND METHODS: Twenty-six young Wistar rats were separated into four groups. On postpartum day 10, six rats received only selenite (group 1, selenite-induced cataract), seven rats received selenite and high dose oral sildenafil (group 2, high-dose sildenafil-treated), seven rats received selenite and low dose oral sildenafil (group 3, low-dose sildenafil-treated), and six rats received only saline (group 4, controls). On postpartum day 30, cataract formation was graded and recorded using an operating microscope. The rats were sacrificed, lens tissues were isolated, and serum samples were collected. Nitrite oxide metabolites (NOx), advanced oxidative protein products (AOPP), and total sulfhydryl (TSH) levels were assessed in both serum and lenticular samples. RESULTS: The rats treated with low-dose sildenafil showed lower levels of AOPP and NOx, and the higher levels of TSH than the rats in other experimental groups. Otherwise, the rats treated with high-dose sildenafil, similar to the selenite-induced cataract group, showed higher levels of AOPP and serum NOx than rats in the low-dose sildenafil-treated group. The rats treated with low-dose sildenafil also showed less cataract development than rats in the other experimental groups. CONCLUSION: Low doses (0.7 mg/kg) of oral sildenafil might show a protective effect on cataract development by lowering oxidative stress.

Effects of ozone preconditioning on recovery of rat colon anastomosis after preoperative radiotherapy.

BACKGROUND: Anastomotic leakage is a devastating complication of colorectal surgery. Neoadjuvant radiotherapy for colorectal cancer can affect the mechanical and biochemical parameters of anastomotic healing. It has been reported that ozone increases antioxidant enzyme activity and stimulates adaptive processes to oppose the pathophysiologic conditions mediated by reactive oxygen species (ROS). OBJECTIVES: The objective of this study was to investigate the effect of controlled administration of ozone on the healing of anastomosis and the activation of antioxidant enzymes in the colon after radiotherapy. MATERIAL AND METHODS: Rats (n = 48) were randomly assigned to the following groups: control groups (1 and 2), saline-treated and irradiated (IR) groups (3 and 4) and ozone oxidative preconditioning (OOP) and IR groups (5 and 6). Rats were exposed to whole-body IR (6 Gy) after pretreatment with either saline or ozone. Rats in groups 1, 3 and 5 were euthanized on postoperative day 3, whereas those in groups 2, 4 and 6 were euthanized on postoperative day 7. The anastomoses were performed on day 7 post-IR. The anastomotic segment was resected to measure hydroxyproline (HPO) content, myeloperoxidase (MPO) activity and malondialdehyde (MDA) concentration and for histopathological evaluation. RESULTS: The mean bursting pressure of the groups that underwent radiotherapy was lower than that of the control groups (p < 0.001). In groups 5 and 6, the tissue HPO concentrations were higher than those in groups 3 and 4. Although mean values for MPO activity in groups 5 and 6 were higher than those in groups 3 and 4, the differences were not significant. Regarding oxidative damage markers, MDA concentrations were significantly lower in group 5 than those in group 3. CONCLUSIONS: In this experimental model, OOP exerted favorable effects on colon anastomotic healing after radiation exposure.

Protein Oxidation Levels After Different Corneal Collagen Cross-Linking Methods.

PURPOSE: To evaluate advanced oxidation protein products (AOPP) levels, superoxide dismutase (SOD) enzyme activity, and total sulfhydryl (TSH) levels in rabbit corneas after different corneal collagen cross-linking (CXL) methods. METHODS: Eighteen eyes of 9 adult New Zealand rabbits were divided into 3 groups of 6 eyes. The standard CXL group was continuously exposed to UV-A at a power setting of 3 mW/cm for 30 minutes. The accelerated CXL (A-CXL) group was continuously exposed to UV-A at a power setting of 30 mW/cm for 3 minutes. The pulse light-accelerated CXL (PLA-CXL) group received UV-A at a power setting of 30 mW/cm for 6 minutes of pulsed exposure (1 second on, 1 second off). Corneas were obtained after 1 hour of UV-A exposure, and 360-degree keratotomy was performed. SOD enzyme activity, AOPP, and TSH levels were measured in the corneal tissues. RESULTS: Compared with the standard CXL and A-CXL groups (133.2 ± 8.5 and 140.2 ± 6.2 µmol/mg, respectively), AOPP levels were found to be significantly increased in the PLA-CXL group (230.7 ± 30.2 µmol/mg) (P = 0.005 and 0.009, respectively). SOD enzyme activities and TSH levels did not differ between the groups (P = 0.167 and 0.187, respectively). CONCLUSIONS: CXL creates covalent bonds between collagen fibers because of reactive oxygen species. This means that more oxygen concentration during the CXL method will produce more reactive oxygen species and, thereby, AOPP. This means that in which CXL method occurs in more oxygen concentration that will produce more reactive oxygen species and thereby AOPP. This study demonstrated that PLA-CXL results in more AOPP formation than did standard CXL and A-CXL.

Serum Copeptin, Pentraxin 3, Anti-Mullerian Hormone Levels With Echocardiography and Carotid Artery Intima-Media Thickness in Adolescents With Polycystic Ovary Syndrome.

BACKGROUND: The aim of the study was to investigate the presence of possible markers in the prediction of polycystic ovary syndrome (PCOS)-related metabolic alterations and cardiovascular events in adolescent PCOS cases and also to investigate the applicability of anti-Mullerian hormone (AMH) levels for the diagnosis of PCOS. METHODS: In this cross-sectional study, a total of 75 non-obese women (adolescent PCOS group, n = 25; adult PCOS group, n = 25; control group, n = 25) were included. Measurements of copeptin, pentraxin 3 (PTX3), and AMH serum levels were performed. RESULTS: Serum copeptin, PTX3 and echocardiographic indices were not significantly different in PCOS subjects and they did not have higher common carotid artery intima-media thickness (CIMT) measurement. AMH levels were significantly higher in PCOS patients. There was a positive correlation between AMH and mean ovarian volume (r = 0.58, P < 0.001) and between AMH and total testosterone level (r = 0.63, P < 0.001). In order to predict a threshold value for the diagnosis of PCOS by using AMH, the receiver operating characteristic (ROC) method was used. Area under the curve was 0.820 and cut-off point was 6.66 ng/mL for AMH with a sensitivity of 62% and specificity of 76%. CONCLUSIONS: Possible markers for PCOS-related metabolic alterations may not present in the adolescent years. Serum AMH may be useful as a diagnostic test for adolescents.

Can Riboflavin Penetrate Stroma Without Disrupting Integrity of Corneal Epithelium in Rabbits? Iontophoresis and Ultraperformance Liquid Chromatography With Electrospray Ionization Tandem Mass Spectrometry.

PURPOSE: To examine riboflavin concentrations in corneas and aqueous humor from rabbits with standard and transepithelial methods and iontophoresis without disrupting the integrity of the corneal epithelium before corneal collagen cross-linking. METHODS: Twenty-four eyes of 12 adult New Zealand rabbits were used. They were assigned to 4 groups, each including 6 eyes. Group 1 was exposed to the standard method and given riboflavin 0.1% after epithelial debridement. Group 2 was exposed to the transepithelial method and given benzalkonium chloride (BAC), ethylenediaminetetraacetic acid (EDTA), trometamol (TRIS), hydroxypropylmethylcellulose (HPMC), and riboflavin 0.2% 3 times at 1.5-minute intervals followed by riboflavin 0.2%. Group 3 was given riboflavin 0.1% by using 1-mA electric current for 10 minutes with the help of iontophoresis without using substances disrupting the integrity of the corneal epithelium. Group 4 received the same treatment as did group 3, except that it was given riboflavin 0.2%. Following these treatments, riboflavin concentrations in aqueous humor and corneas were measured with ultraperformance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). RESULTS: Riboflavin concentrations in the cornea and aqueous humor were higher in group 1 (42.4 ± 5.4 µg/g) than in the other groups. They were significantly higher in group 4 (34.2 ± 6.6 µg/g) than in group 2 (24.4 ± 1.2 µg/g) (P = 0.009) and group 3 (23.6 ± 6.1 µg/g) (P = 0.026). There was not a significant difference in corneal riboflavin concentrations between group 2 and group 3 (P = 0.937). CONCLUSIONS: Intrastromal and aqueous riboflavin concentrations after administration of riboflavin 0.2% through iontophoresis without disrupting the integrity of the corneal epithelium were lower than those after the standard method, but higher than those after the transepithelial method. In this study, in which riboflavin concentrations were measured with a very sensitive method, iontophoresis was observed to increase the transmission of riboflavin molecules into the cornea without using substances disrupting epithelial integrity.

Sirtuin gene polymorphisms are associated with chronic obstructive pulmonary disease in patients in Mugla province.

INTRODUCTION: Chronic obstructive pulmonary disease (COPD) is an irreversible progressive chronic inflammatory disease that causes shortness of breath in consequence of a decrease in pulmonary functions. The pulmonary inflammatory pathogenesis is multifactorial. We have too little up-to-date information about the relation between COPD and genetics. In our study, the relation with the SIRT1 gene's mononucleotide polymorphisms (SNP) rs7895833, rs7069102 and rs2273773 was analyzed through various laboratory data. MATERIAL AND METHODS: One hundred COPD patients from the archive records of the Chest Diseases Department of Mugla Sitki Kocman University Medical Faculty were included in the study. A control group was constituted from 100 healthy individuals who live in the same geographical region. The SIRT1 genotypes for these patients were determined using polymerase chain reaction (PCR) and confronting two-pair primers (CTPP) methods. The SIRT1 gene polymorphisms rs7895833, rs7069102 and rs2273773 were analyzed. GG, AG, AA genotypes and G and A alleles of rs7895833, TT, TC, CC genotypes and T and C alleles of rs2273773, and CC, CG, GG genotypes and C and G alleles of rs7069102 were examined. The data in both groups were compared. CONCLUSIONS: A significant difference between GG, AG and AA genotypes of rs7895833 was found. Especially, the AG genotype was observed more in the group with COPD, with a significant difference. A significant difference between TT, TC and CC genotypes of rs2273773 was found. There was a significant difference between two groups with regards to C and G alleles of rs7069102. A significant difference was not found between the groups with regards to G and A alleles of rs7895833. A difference was not found for both groups between T and C alleles of rs2273773. It shows that these polymorphisms of the SIRT1 gene may be associated with COPD.

Effects of ozone oxidative preconditioning on liver regeneration after partial hepatectomy in rats.

AIM: Similar protective effect of ischemic and ozone oxidative preconditioning (OzoneOP) in hepatic ischemia-reperfusion (I/R) injury was demonstrated, providing evidences that both preconditioning settings shared similar biochemical mechanisms of protection. We investigated the effects of OzoneOP on liver regeneration after 70% partial hepatectomy (PHx) in rats. METHODS: Rats were divided into three groups: PHx, I/R + PHx, and OzoneOP + I/R + PHx groups. Ozone (intraperitoneal, 1.2 mg/kg) was given to rats subjected to I/R and 70% hepatectomy daily five times before operation. At 24 hr and 48 hr after resection, samples were collected for the measurement of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor alpha (TNF-α), and interleukin-6 (IL-6). Moreover, liver regeneration rate, proliferating cell nuclear antigen (PCNA) labeling index, mitotic index, and histopathological examination were evaluated. RESULTS: OzoneOP reduced liver injury determined by liver histology and serum transaminases. There was a rise in serum TNF-α and IL-6 levels in the I/R + PHx group whereas OzoneOP significantly decreased the rise in the level of TNF-α but not IL-6 on the 24 hr and 48 hr of reperfusion. Moreover, liver regeneration in OzoneOP + PHx group, as assessed by the regenerated liver weight, mitotic, and PCNA-labeling index, was significantly improved when compared to I/R + PHx group. CONCLUSION: These results suggest that OzoneOP ameliorates the hepatic injury associated with I/R and has a stimulatory effect on liver cell regeneration that may make it valuable as a hepatoprotective modality.

N-acetyl-cysteine improves anastomotic wound healing after radiotherapy in rats.

AIM: This study was designed to determine the effects of intraperitoneally or orally administered N-acetylcysteine (NAC) on anastomotic healing of irradiated rats. METHODS: Thirty-two male Wistar albino rats were randomized into four groups containing 8 rats each: I; standard resection plus anastomosis, II; radiation plus standard resection plus anastomosis, III; radiation plus standard resection plus anastomosis plus oral NAC, IV; radiation plus standard resection plus anastomosis plus intraperitoneal NAC. Four types of assessment were performed: bursting pressure, hydroxiproline (OHP) content, histopathology, and biochemical evaluation, including serum malondialdehyde (MDA), advanced oxidation protein products (AOPP), reduced glutathione (GSH) and superoxide dismutase (SOD) activities. RESULTS: Group comparisons demonstrated that bursting pressure was significantly higher in NAC treated rats. The mean tissue OHP concentration in the anastomotic tissue was significantly lower in irradiated rats (group II) than in the other groups. NAC treatment caused increased activity of SOD and GSH. In contrast, MDA levels were found to be decreased in groups III and IV. Histopathological analysis revealed that NAC administration, either orally or intraperitoneally, leads to a better anastomotic healing in terms of reepithelialization, perianastomotic fibrosis, ischemic necrosis, and muscle layer destruction. CONCLUSION: The present study supports the hypothesis that NAC administration alleviates the negative effects of radiotherapy on anastomotic healing. Nevertheless, the underlying mechanisms responsible for this protective effect is unknown today.

Effects of erdosteine on cyclosporine-A-induced hepatotoxicity in rats.

Cyclosporine A (CsA) is a potent immunosuppressive agent used for organ transplantations and various autoimmune disorders. However, hepatotoxicity due to CsA remains one of the major side effects. The use of antioxidants reduces the adverse effects of CsA. The aim of this study was to determine the protective effects of erdosteine on CsA-induced liver injury through tissue oxidant/antioxidant parameters and to evaluate light microscopic alterations in rat-liver tissues. Rats were randomly divided into four experimental groups: The control group received sunflower oil (2 mL/kg/day, per orally; p.o.), while the other groups were treated with CsA (25 mg/kg/day, p.o.) or erdosteine (10 mg/kg/day, p.o.) or CsA+erdosteine, respectively. Serum aspartate aminotransferase and alanine aminotransferase levels, tissue malondialdehyde and nitric oxide levels, and superoxide dismutase, glutathione peroxidase and catalase enzyme activities were measured. Histological examination was performed. CsA caused a significant deterioration in the hepatic function tests, morphology, and gave rise to severe oxidative stress in the liver. Erdostein significantly improved the functional and histological parameters and attenuated the oxidative stresss induced by CsA. Erdostein protects liver tissue against oxygen free radicals and prevents hepatic dysfunction and morphological abnormalities associated with chronic CsA administration.

Effects of CO(2) pneumoperitoneum on anastomotic healing in rats receiving preoperative 5-fluorouracil neoadjuvant chemotherapy.

BACKGROUND: When used separately, antineoplastic agents and carbon dioxide (CO(2)) pneumoperitoneum have been reported to impair anastomotic healing in experimental animals. However, the effects of their combined use have not been previously investigated. The aim of this study was to investigate the possibility that neoadjuvant chemotherapy with 5-fluorouracil followed by CO(2) pneumoperitoneum would affect the healing of anastomoses in the colon. METHODS: Sprague-Dawley rats (n = 48) were given 5-fluorouracil (20 mg/kg/day) for 5 days, and were then assigned to one of the three groups. Prior to surgery, the control group received no pneumoperitoneum. The other two groups received pneumoperitoneum at 6 and 12 mmHg, respectively, for 2 hr. The large intestine was transected and anastomosis was performed via median laparotomy. On postoperative days 3 and 7, relaparotomy was performed in half of the rats in each group. From the colon, a segment including the anastomosis was excised. Tissue hydroxyproline levels were measured. For histological evaluation, the Verhofstad scale was modified and used. RESULTS: No significant differences in hydroxyproline levels were seen across the groups on postoperative days 3 or 7. However, by postoperative day 7, polymorphonuclear leukocytes and necrosis in the 6-mmHg group had decreased markedly, and granulation had improved. CONCLUSION: Overall, these findings suggest that preoperative 5-fluorouracil therapy followed by pneumoperitoneum at 6 or 12 mmHg does not impair anastomotic healing.

A possible mechanism for transpupillary thermotherapy: nitric-oxide-related cellular damage.

BACKGROUND: To determine the oxidative stress markers on rabbit vitreous following transpupillary thermotherapy (TTT) application. METHODS: The pigmented rabbit eyes were divided into 3 groups, each containing 6 eyes. Group 1 was used as a control group. Twelve eyes underwent TTT with a power of 250 mW (group 2) and 800 mW (group 3), with a diameter of 3000 microm and duration of 60 s; 24 h after laser application, vitreous samples were collected. Nitric oxide (NO) and malondialdehyde (MDA) levels and superoxide dismutase (SOD) activities were determined in all groups. RESULTS: NO levels were statistically significantly higher in all groups when compared with the control group (p < 0.05). MDA levels were higher and SOD activities were lower in all groups when compared with the control group, but the differences were not significant statistically (p > 0.05). INTERPRETATION: Our results support the hypothesis that TTT application induces NO synthesis, which may lead to occlusion at choroidal neovessels. Because of the nonsignificantly increased levels of MDA and decrease of SOD activities there maybe only a weak relation between lipid peroxidation induced by free oxygen radicals and TTT-induced vascular damage.

Carnosine may reduce lung injury caused by radiation therapy.

Ionising radiation is known one of the most effective tools in the therapy of cancer but in many thoracic cancers, the total prescribed dose of radiation that can be safely administered to the target volume is limited by the risk of complications arising in the normal lung tissue. One of the major reasons for cellular injury after radiation is the formation of reactive oxygen species (ROS). Radiation pneumonitis is an acute phase side-effect which generally subsides after a few weeks and is followed by a chronic phase characterized by inflammation and fibrosis, that can develop months or years after irradiation. Carnosine is a dipeptide composed by the amino acids beta-histidine and l-alanine. The exact biological role of carnosine is not totally understood, but several studies have demonstrated that it possesses strong and specific antioxidant properties, protects against radiation damage,and promotes wound healing. The antioxidant mechanism of carnosine is attributed to its chelating effect against metal ions, superoxide dismutase (SOD)-like activity, ROS and free radicals scavenging ability . Either its antioxidant or anti-inflammatuar properties, we propose that carnosine ameliorates irradiation-induced lung injury. Thus, supplementing cancer patients to whom applied radiation therapy with carnosine, may provide an alleviation of the symptoms due to radiation-induced lung injury. This issue warrants further studies.