Carvacrol protects rat liver exposed to formaldehyde by regulating oxidative stress, and asprosin and subfatin hormones.

Toxic doses of formaldehyde (FA) can cause oxidative damage and impair energy metabolism. Asprosin (ASP) and subfatin (SUB) are adipokines produced by adipose tissue that help regulate energy metabolism. We investigated the effects of carvacrol (CAR), an antioxidant with hepatoprotective properties, on ASP and SUB in rats exposed to FA using immunohistochemistry and biochemistry. We used 42 male Wistar albino rats divided into six groups of seven: group 1, untreated control; group 2, FA (10 ppm FA by inhalation 8 h/day, 5 days/week); group 3, CAR-20 (20 mg/kg); group 4, CAR-40; group 5, FA (10 ppm FA by inhalation 8 h/day, 5 days/week) + CAR-20 (20 mg/kg); group 6, FA (10 ppm FA by inhalation 8 h/day, 5 days/week) + CAR-40 (40 mg/kg). Levels of ASP and SUB, and total oxidant status (TOS) and total antioxidant status (TAS) in blood and liver tissue were measured using ELISA. ASP and SUB immunoreactivity was assessed using immunohistochemistry. The number of apoptotic cells was determined using the TUNEL method. The number of apoptotic cells in group 2 was increased compared to group 1. TOS in group 2 was increased compared to group 1. The numbers of apoptotic cells and TOS in group 3 were decreased compared to group 1. TOS was decreased in group 6 compared to group 2, but TOS was increased compared to group 1. We found ASP and SUB immunoreactivity in the liver. All alterations were reversed by addition of CAR. It appears that FA disrupts energy metabolism and CAR ameliorates the destructive effects of FA when used at appropriate doses, although CAR might be harmful at high doses.

Irisin, Angtpl8, Elabela and antioxidants alteration in rats with and without energy drink and treadmill exercise.

OBJECTIVE: In this study, we sought to investigate the effects of energy drink supplementation and treadmill exercise on the levels of free radicals, antioxidants, Angtpl8, Elabela, and lipid metabolism in rats. MATERIALS AND METHODS: A total of 28 male Wistar albino rats (4 weeks old, 101.96 ± 9.75 g) were included in the study. The rats were randomly divided into four equal groups: control, exercise, supplement, and exercise+supplement groups. At the end of the study, the rats were decapitated, and blood samples were tested for levels of Angptl-8, ghrelin, leptin, irisin, SOD, CAT, TBARS, total oxidant status, and total antioxidant status using enzyme-linked immunosorbent assay. Levels of blood lipids including triglycerides, total cholesterol, HDL-C, and LDL-C were studied using spectrophotometric method in an auto analyzer. RESULTS: Statistical analysis showed statistical significance in TBARS, LDL-C, irisin, Angptl-8, and Elabela levels of the exercise group; SOD and HDL-C levels of the supplement+exercise group; and total cholesterol levels in the supplement group (p < 0.05). Although there were differences between the groups in leptin, ghrelin, and CAT levels, they were not statistically significant (p > 0.05). CONCLUSIONS: As a result, it can be argued that treadmill exercise is important in regulating lipid metabolism and stimulating peptide hormones and receptors. Furthermore, consuming energy drinks without performing exercise or physical activity increases fat stores, and such increases in the critical organs and tissues may pose a threat to the body.

Effects of carnosine on apoptosis, transient receptor potential melastatin 2, and betatrophin in rats exposed to formaldehyde.

We investigated the effects of exposure to formaldehyde on transient receptor potential melastatin 2, betatrophin, total oxidant status and total antioxidant status in rat liver and kidney tissues. We also investigated the effects of carnosine on formaldehyde treated animals. We used 28 male rats divided ramdomly into four groups of seven: untreated control group, carnosine treated group, formaldehyde treated group and formaldehyde + carnosine group. The experiment lasted for four weeks. Betatrophin levels in samples were measured uing the enzyme-linked immunosorbent assay, and total oxidant status and total antioxidant status were measured using REL assay diagnostic kits. We detected betatrophin and transient receptor potential melastatin 2 immunoreactivity using immunohistochemistry and assessed apoptosis using terminal deoxynucleotidyl transferase dUTP nick end labeling. The betatrophin and total antioxidant status levels decreased in kidney, liver and plasma following exposure to formaldehyde, while total oxidant status and terminal deoxynucleotidyl transferase dUTP nick end labeling positivity increased. Carnosine supplementation reversed histopathology and biochemical damage caused by formaldehyde. We suggest that carnosine treatment may be useful for protecting persons exposed to formaldehyde.

Immunostaining characteristics of irisin in benign and malignant renal cancers.

We investigated the expression of irisin in renal cancers using immunocytochemistry. Irisin has been reported to exhibit anticancer properties. The study groups consisted of 22 cases each of control renal tissue, oncocytoma, chromophobe renal cell carcinoma (RCC), clear cell RCC (Fuhrman nuclear grades 1, 2, 3 and 4) and papillary RCC. We evaluated 10 slides for each of 176 cases. Slides were immunostained for irisin and histoscores were calculated for the prevalence and strength of immunostaining. Fuhrman nuclear grade 1, 2, 3 clear cell RCC and papillary RCC exhibited no irisin immunoreactivity. Irisin immunoreactivity was observed in some Fuhrman nuclear grade 4 RCCs. We found a significant decrease in irisin staining in chromophobe RCC compared to the control. Immunoreactivity in the oncocytoma tissue was comparable to the control group. Irisin immunoreactivity in chromophobe RCC decreased and no immunoreactivity was observed in Fuhrman nuclear grade 1, 2, 3 clear cell RCC and papillary RCC. Immunistochemical screening of irisin in renal oncocytomas and renal cancers may be useful for differential diagnosis.

A promising biomarker to distinguish benign and malignant renal tumors: ELABELA.

AIM: The aim of this study was to investigate ELABELA (ELA) expression in benign and malignant renal tissues and expression differences in different nuclear grades of clear cell carcinomas. MATERIALS AND METHODS: Patients that underwent surgery due to renal masses between the years of 2007 and 2017 were used. Control renal tissues (n = 23), papillary RCC (n = 23), clear cell RCC (CcRCC) [Fuhrman Grade1 (n = 23), Fuhrman Grade2 (n = 23), Fuhrman Grade3 (n = 23), Fuhrman Grade4 (n = 23)], and chromophobe RCC (n = 23) were included to the study. The Independent samples t-test was used for 2-point intergroup assessments and the one-way analysis of variance and posthoctukey test was used for the others. Values of P < 0.05 were considered statistically significant. RESULTS: ELA immunoreactivity was observed in proximal and distal tubules in the kidney, but not in glomeruli in control tissues. When compared with control kidney tissue, a statistically significant increase was observed in ELA immunoreactivity in renal oncocytoma. In the chromophobe RCC, ELA immunoreactivity was significantly lower than control kidney tissue, whereas papillary RCC did not show ELA immunoreactivity. However, compared with control kidney tissue, ELA immunoreactivity was not observed in Fuhrman Grade 1 and Grade 2 CcRCC. Also, there was a significant decrease at Fuhrman Grade 3 and Grade 4 CcRCC compared with control kidney tissues. In the statistical analysis of ELA immunoreactivity among the Fuhrman nuclear grades of CcRCCs, The ELA immunoreactivity was higher at Grade 4 CcRCC than Grade 1, Grade 2, and Grade 3. CONCLUSION: ELA is a usefull molecule to differentiate benign and malign renal tumors. But further broad and comprehensive studies are needed to investigate cellular and molecular mechanisms of ELAs on malign transformation.

Ghrelin and NUCB2/Nesfatin-1 expression in unilateral testicular torsion-induced rats with and without N-acetylcysteine.

Testicular torsion (TT) is a common urological problem in the field of pediatric surgery. The degree and duration of torsion determines the degree of testicular damage; however, its effects on the expression of octanoylated ghrelin and nucleobindin 2 (NUCB2) /nesfatin-1 synthetized from testicular tissue remain unclear. We explored the effects of experimentally induced unilateral TT on serum and contralateral testicular tissue ghrelin and NUCB2/nesfatin-1 levels, and determined whether N-acetyl cysteine (NAS) treatment had any effects on their expression. A total of 42 Wistar Albino strain rats were divided into 7 groups: Group (G) I control, GII sham, GIII 12-hour torsion, GIV 12-hour torsion + detorsion + 100 mg/kg NAS, GV 24-hour torsion, GVI 24-hour torsion + detorsion + 100 mg/kg NAS, and GVII 100 mg/kg NAS. Octanoylated ghrelin and NUCB2/nesfatin-1 concentrations were evaluated in serum using the ELISA method and in testicular tissue with immunohistochemical methods. Immunoreactivity of octanoylated ghrelin significantly increased in GI compared to GIII, GV, and GVI (p<0.05). NUCB2/nesfatin-1 immunoreactivity increased in GV and GVIII relative to GI (p<0.05). In the 12-hour torsion group, a significant decrease in octanoylated ghrelin levels with NAS treatment was observed; however, in the 24-hour torsion group, a significant decrease was not observed. In the 12-hour torsion + NAS treatment group, a significant change was not observed in NUCB2/nesfatin-1 expression. Following 24-hour torsion, an increase in NUCB2/nesfatin-1 levels was observed, and NAS treatment did not reverse this increase. It was determined that increases in the expression of octanoylated ghrelin and NUCB2/nesfatin-1, the latter of which was a result of TT, reflect damage in this tissue. Importantly, NAS treatment could prevent this damage. Thus, there may be a clinical application for the combined use of NAS and octanoylated ghrelin in preventing TT-related infertility.

Irisin immunostaining characteristics of breast and ovarian cancer cells.

To determine expression pattern of irisin in tissues obtained from human ovarian cancer, breast cancer, and cervix cancer. Tissue samples obtained from subjects with breast cancer, ovarian cancer cervix cancer, simple endometrial hyperplasia, complex atypical endometrial hyperplasia. At least five sections from each subject were immunohistochemically stained with irisin antibody, and H-score method was used to evaluate irisin intensity. Tissues obtained from healthy breast tissues, proliferative phase endometrium adenomyosis and benign ovarian tumors were accepted as control. Irisin activity was not detected in control breast tissues significantly increased irisin staining was detected in invasive lobular, intraductal papillary, invasive ductal, invasive papillary, and mucinous carcinomas compared to control tissues. Also, significantly increased irisin immunoreactivity was detected in both ovarian endometriosis and mucinous carcinomas compared to benign tumors. However irisin staining was not observed at the papillary carcinoma of the ovary while sections obtained from simple and complex atypical endometrial hyperplasia, and cervix carcinoma demonstrated irisin immunoreactivity. Increased irisin immunoreactivity in tissues obtained from breast, ovary, cervix carcinomas, and endometrial hyperplasia suggest critical role of this peptide during carcinogenesis.

Irisin immunohistochemistry in gastrointestinal system cancers.

Cancer is the leading cause of morbidity and mortality worldwide. Some studies have shown that high heat kills cancer cells. Irisin is a protein involved in heat production by converting white into brown adipose tissue, but there is no information about how its expression changes in cancerous tissues. We used irisin antibody immunohistochemistry to investigate changes in irisin expression in gastrointestinal cancers compared to normal tissues. Irisin was found in human brain neuroglial cells, esophageal epithelial cells, esophageal epidermoid carcinoma, esophageal adenocarcinoma and neuroendocrine esophageal carcinoma, gastric glands, gastric adenosquamous carcinoma, gastric neuroendocrine carcinoma, gastric signet ring cell carcinoma, neutrophils in vascular tissues, intestinal glands of colon, colon adenocarcinoma, mucinous colon adenocarcinoma, hepatocytes, hepatocellular carcinoma, islets of Langerhans, exocrine pancreas, acinar cells and interlobular and interlobular ducts of normal pancreas, pancreatic ductal adenocarcinoma, and intra- and interlobular ducts of cancerous pancreatic tissue. Histoscores (area × intensity) indicated that irisin was increased significantly in gastrointestinal cancer tissues, except liver cancers. Our findings suggest that the relation of irisin to cancer warrants further investigation.

Alteration of serum and cardiac tissue adropin, copeptin, irisin and TRPM2 expressions in DOX treated male rats.

Doxorubicin (DOX) cardiotoxicity is a significant side effect in cancer survivors. DOX and its metabolites alter cardiac gene expression and affect metabolic energy-related peptides. Adropin, copeptin, irisin and TRPM2 are produced locally in the heart and play a role in energy homeostasis. We investigated the fates of adropin, copeptin, irisin and TRPM2 in serum and cardiac tissues of DOX treated rats. Animals were divided into three groups of six: 1) untreated controls, 2) DOX treated and 3) saline treated. The rats were fed a standard diet ad libitum for 14 days then were sacrificed and heart and serum samples were taken. Adropin, copeptin, irisin levels in tissue homogenates and serum were measured using ELISA. Immunoreactivity of heart tissue adropin, copeptin, irisin and TRPM2 also were investigated. The peptides increased in both serum and cardiac tissue homogenates in animals treated with DOX compared to the other groups. DOX increased adropin in endocardial and myocardial cells, but it decreased expression of copeptin. DOX did not affect endocardial irisin and TRPM2 expressions, but myocardial irisin and TRPM2 expressions were increased. Serum adropin, irisin and copeptin were increased in DOX treated rats. Cardiac adropin, copeptin, irisin and TRPM2 are affected by DOX and may play a role in DOX cardiotoxicity.

Immunohistochemical expressions of adropin and inducible nitric oxide synthase in renal tissues of rats with streptozotocin-induced experimental diabetes.

Diabetes is characterized by high blood glucose levels; it occurs in 30-35% of the population. Elevated glucose levels can damage a number of organs, including the kidneys. Several peptide hormones participate in maintaining glucose homeostasis including the recently discovered "adropin," a 42 amino acid peptide hormone. Adropin also alters inducible nitric oxide synthase (iNOS) expression. Therefore, we studied how adropin and iNOS expression is altered in the renal tissues of streptozotocin (STZ) induced diabetic rats. Seven sham, seven control and seven Wistar albino male rats were fed standard rat pellets and water ad libitum for 10 weeks. The rats in the diabetic group were injected i.p. with a single dose of 60 mg/kg STZ dissolved in 0.1 M phosphate-citrate buffer, pH 4.5. After the 10-week experimental period, the rats in both groups were anesthetized and decapitated. Kidney tissues were excised and placed in 10% formaldehyde solution, taken through routine histological procedures, and embedded in paraffin. Sections 5-6 µm thick were stained immunohistochemically using the avidin-biotin complex (ABC) method. Adropin and iNOS immunoreactivity were co-localized in the glomeruli, peritubular interstitial cells and peritubular capillary endothelium of the cortex; the thin limb of the loop of Henle in the medulla; and medullary peritubular interstitial cells and endothelium of the peritubular capillaries in both the control and diabetic groups. The intensities of adropin and iNOS immunoreactivity increased with the severity of the diabetes. Intense adropin immunoreactivity was detected in both the smooth muscle and human small intestine Paneth cells that were used as positive controls. The elevated levels of adropin and iNOS in the kidney indicates that these substances are involved in the pathophysiology of diabetes; this constitutes a compensatory mechanism against the damage inflicted by the disease.

The effects of different prosthetic materials on the formation of collagen types in incisional hernia.

OBJECTIVE: The aim of this study was to determine the rate of collagen Type I/Type III for different meshes. METHOD: Fifty rats were used. Five groups were formed: prolene (n = 10), mersilene (n = 10), parietex (n = 10), e PTFE (n = 10) and control group (n = 10). In all animals, laparotomy was performed using a midline incision. After that four different kinds of meshes are placed into the retro-rectus plane and fixed with a non-absorbable suture. Rectus superficial fascia and skin are closed. In the control group, repairment is done primarily. Thirty days later, meshes are found through the incisions that were done previously. Scar tissues above and near by meshes and also in the control group are taken, and in these tissue samples, the ratio of Type I/III is evaluated histochemically. RESULTS: The Prolene mesh was found to contain more collagen fibers than e PTFE. As a result of the histopathologic evaluation, it was seen that Group I contained statistically significantly more collagen density than the other four groups (p < 0.05). Moreover, the collagen Type I/III ratio in the specimen taken from the top part and the surrounding area of Group I was found significantly higher than the collagen Type I/III ratios of the rest of the groups (p < 0.05). CONCLUSION: As a conclusion, the ratio of collagen Type I/III is the highest in the prolene group.

Effects of melatonin and vitamin C on cigarette smoke-induced damage in the kidney.

This study was carried out to investigate smoke-induced structural and biochemical changes and protective effects of co-administered melatonin and vitamin C in the kidney. Twenty-four Wistar adult female rats were used in this study. Animals were divided into four groups. The first group rats were used as control. The second group of rats inhaled cigarette smoke. Smile smoke inhaling third and fourth group rats received melatonin and vitamin C, respectively. At the end of experimental study, kidney tissues and blood samples were taken under ether anesthesia. Tissues were prepared and examined by light microscopy. Malondialdehyde and glutathione levels and catalase activity were determined. By light microscopic observation, a decrease of Bowman space of some renal corpuscles, foamy-like tubules, dilatation and congestion of the peritubuler vessels, and atrophy of the some renal corpuscles were observed in group II. In groups III and IV melatonin and vitamin C relatively protected the kidney tissue against smoke intoxication. Biochemical examination showed that malondialdehyde and glutathione levels and catalase activity in group II were higher than in group I. Melatonin and vitamin C injection to group III and IV caused a decrease in malondialdehyde and glutathione levels. Catalase activity did not change in these groups. We have shown that cigarette smoke inhalation caused structural changes in the kidney. However, melatonin and vitamin C administration produced in some degree protection against smoke-induced damage.