Vitamin D supplementation alleviates diabetic complications by increasing the amount of irisin in testicular tissues and blood of rats with experimental diabetes.

OBJECTIVE: Diabetes is an important endocrinological disease that has an increasing incidence in the world and affects all biological tissues including testicles. Therefore, this study aimed to reveal the histological and biochemical effects of vitamin D on irisin, apoptosis, total antioxidant status (TAS), and total oxidant status (TOS) in testicular tissues of rats with experimental diabetes. MATERIALS AND METHODS: 41 male Wistar rats, 8-10 weeks old, weighing between 200-220 g, were included in the study as the following groups: control group (n=7; no treatment), sham group [only sodium citrate buffer (SCB)] [n=7; single dose 0.1 Molar (M) SCB given intraperitoneally (i.p)], vitamin D group (n=7; 50 IU/day given orally), diabetes group [n=10; single dose 50 mg/kg Streptozotocin (STZ) dissolved in 0.1 M SCB and given i.p (tail vein blood glucose level above 250 mg/dl after 72 hours)] and diabetes+vitamin D group [n=10, single dose 50 mg/kg STZ, dissolved in 0.1 M SCB and given i.p (tail vein blood glucose level above 250 mg/dl after 72 hours) and when diabetes occurs, oral vitamin D administration of 50 IU/day)]. At the end of the 8 weeks experiment, blood was drawn from the tail vein of all rats, they were sacrificed and testicular tissues were taken. While the amount of irisin in the blood and testicular tissue supernatants was analyzed with the Enzyme-Linked Immunosorbent Assay (ELISA) method, TAS and TOS measurements were analyzed with the REL method, testicular tissues were analyzed histopathologically, immunohistochemically, and with the TUNEL method. RESULTS: When the diabetes group was compared with the control and sham groups, it was reported that the amounts of blood and tissue supernatant irisin and TAS significantly decreased and the TOS was significantly increased; a statistically significant increase in irisin and TAS of blood and tissue supernatants and a significant decrease in TOS were detected when diabetes+vitamin D and diabetes groups were compared among themselves. Similar results were obtained in the immunohistochemical studies. Tissue expressions of irisin decreased in the diabetes group compared to the control and sham groups, while the application of vitamin D increased the tissue expressions of irisin. Additionally, when the numbers of apoptotic cells were compared, it was reported that apoptotic cells in the diabetes group increased significantly compared to the control and sham groups, and vitamin D administration significantly decreased the number of apoptotic cells. CONCLUSIONS: Taken together, vitamin D administration to diabetic rats decreased the number of apoptotic cells and increased the amount of irisin. Vitamin D had an effective role in maintaining the physiological integrity of rat testicular tissues, so vitamin D may be a potent agent to be used in the treatment of diabetes in the future.

Histopathological, immunohistochemical and biochemical evaluation of electroacupuncture treatment of nervus radialis and nervus ulnaris injuries in rabbits.

The aim of this study was to evaluate the efficacy of electroacupuncture in acute and chronic phases of radial and ulnar nerve injuries in histopathological, immunohistochemical and biochemical aspects. In the study, the rabbits were divided into four groups namely acute nerve injury (ANI) group, chronic nerve injury (CNI) group, positive control (PC) group and negative control (NC) group. In the ANI, CNI and PC groups, damage was created on the nervus radialis and nervus ulnaris by applying pressure for 60 seconds using a hemostatic forceps under anesthesia. No damage was created in the NC group. Fifteen sessions of electroacupuncture were applied to the rabbits in the ANI, CNI, and NC groups every other day using LI-4 (Large Intestine Meridian-4, He Gu), LI-10 (Large Intestine Meridian-10, Shou San Li), LR-3 (Liver Meridian-3, Tai Chong), and ST-36 (Stomach Meridian-36, Zusanli) electroacupuncture points. Electroacupuncture was not applied to the rabbits in the PC group. Decapitation was performed under general anesthesia at the end of electroacupuncture applications. After the euthanasia procedure, the samples obtained were evaluated for histopathological, immunohistochemical and biochemical parameters. In conclusion, degenerative foci in the treatment groups were found to be fewer than in the PC group whereas NGF and S-100 immunoreactivity were higher in the treatment groups than in the PC group. Whereas no statistically significant difference was observed between the treatment groups and the NC group in terms of oxidative stress factors, there was a statistically significant difference between the treatment groups and the PC group. In light of all these data, we have concluded that electroacupuncture is an effective treatment method for peripheral nerve injuries.

Identification of immunohistochemical localization of irisin in the dwarf hamster (Phodopus roborovskii) tissues.

Irisin is mainly secreted by heart and skeletal muscle cells. It is an exercise-induced protein that converts white adipose tissue to brown. Increased irisin expression was lead to weight loss and improved glucose tolerance. We investigated irisin immunoreactivity in various tissues of the dwarf hamsters (Phodopus roborovskii). Tissues were processed, embedded in paraffin, sectioned at 5 µm and stained immunohistochemically for irisin. In the retina, irisin was found almost all layers, except outer nuclear layer. Also, irisin immunoreactivity was observed in the skin, cornea, striated muscle, parotid gland, tongue, oesophagus, stomach and small intestine. The findings from this study support the notion that skeletal muscle is not the primary source of irisin.

Equine goiter associated with excess dietary iodine.

Naturally occurring goiter cases are described in 2 newborn Arabian foals whose mares were supplemented with excess iodine during the final 24 w of the pregnancy. Six nursing foals and 2 mares were also affected clinically with thyroid hypertrophy. At least 12 times the maximum tolerable level of iodine supplementation was given, as the daily iodine intake for each mare was 299 mg. The prevalence of goiter cases was 2 and 9% in the mares and foals, respectively.

Toxicity of dietary Heliotropium circinatum to rats.

Pyrrolizidine alkaloid intoxication was produced in adult, male rats by feeding different levels (0, 1, 3, 5 or 10%) of Heliotropium circinatum for 20 w. Combined GC-MS revealed 0.15% total alkaloid content in the plant material of which 12% and 88% were basic and N-oxide forms, respectively. The specific alkaloids identified were europine (67.33%), heliotrine (16.34%), lasiocarpine (8.12%), heleurine (4.18%), echinatine (1.56%), 7-angeylheliotrine (1.19%), and an unknown alkaloid (1.28%). Neither mortality nor significant clinical changes occurred in test groups. Mild to moderate, dose-related hepatic megalocytosis was the most prominent histopathological finding. In addition to chronic hepatotoxicity, notable medial thickening occurred in the pulmonary arterioles and arteries of the high-dosed groups. This study indicated that H. circinatum plant has limited toxic potential in rats with mild to moderate histological changes and no mortality at the dosing levels, total doses, or time of exposure employed.

Toxicity of Senecio vernalis to laying hens and evaluation of residues in eggs.

Diets with 0.0, 0.5,2.0, or 4.0% ground-aerIal parts of Senecio vernalis were fed to groups of 10 laying hens for 210 d. Plant alkaloid content was 0.14% with 8.57% in the basic form and 91.43% in the N-oxide form. Specific alkaloids were senecionin (66.65%), senecivernin (10.37%), seneciphylline (8.51%), integerrimine (8.44%), retrorsine (3.03%), senkirkine (2.35%) and hydroxysenkirkine (0.65%). At the end of the feeding period, no mortality or clinical signs occurred in any test group. However, decreases in egg production, feed efficiency, feed intake and body weight occurred on diets containing 2 or 4% of the plant. Serum gamma-GT was significantly elevated and serum albumin and protein were significantly decreased in hens fed the 2 and 4% diets. Total bilirubin was significantly higher in the 4% group. No free pyrrolizidine alkaloids were detected in eggs indicating that at these rates of dietary exposure, they did not produce residues at the level of detection or that they were bound irreversibly to egg proteins. Hens fed 2 or 4% plant diets had mild to moderate chronic liver changes of periportal or septal fibrosis, megalocytosis, bile duct hyperplasia and early regenerative nodule formation.

Toxicity of dietary Heliotropium dolosum seed to Japanese quail.

Five groups of 30 male Japanese quail, each 7-w-old, were fed diets containing 0, 1, 3, 5, or 10% (w/w) of dehulled H dolosum seed. Half of the birds from the each group were killed at 6 and 24 w after beginning of the trial. At the end of 6th w, neither mortality nor clinical sign occurred in test groups. In the 5 and 10% inclusion levels, mild to moderate hepatic injury was detected as evidenced by mild karyomegaly, moderate fatty change, focal or portal fibrosis, bile duct hyperplasie, and ovalocyte proliferation along with lower serum protein and albumin levels. By the termination of the experiment (24 w), 5 birds died in the 10% dosed group. Hepatic cirrhosis was the most prominent finding in the 5 and 10% group; at these levels, serum protein and albumin values decreased significanty while billuribin and ALP levels increased. Based on relative weights and histological evaluations, testicularatrophywasdosedependent. These results partly affirm earlier studies that the quail is highly resistant to the toxic effects of H dolosumseed. However, the resistance to H dolosum is less than Senecio Jacobae and Crotalaria spectabilis when considering tissue injury.

Toxicity of dietary Heliotropium dolosum seed to mice.

Experimental pyrrolizidine alkaloid intoxication was produced in inbred Swiss mice. Animals were fed diets containing 0, 1, 3, 5, or 10% Heliotropium dolosum seed for 24 w. The seeds contained 0.13% total alkaloid concentration composed of 4 specific components: lasiocarpine (78.79%), heliosupine (11.96%), echimidine (5.43%), and heliotrine (3.82%). Deaths occurred in all dosed groups and increased with dietary seed concentration. Massive to submassive liver necrosis together with sinusoidal congestion, and hemorrhage or multifocal hepatocytic necrosis was limited to animals which survived < 5 w and were fed on 10% seed. Moderate to severe hepatomegalocytosis, scattered single cell necrosis, and mild subcapsuler fibrosis were seen in all test group animals that survived > 5 w. Intranuclear eosinophilic inclusions in hepatocytes and bile duct and ductular cell hyperplasia were the most noticeable lesions in the 1, 3, and 5% groups. There was mild to moderate renal tubular megalocytosis in the 3, 5, and 10% groups. It seems likely that H dolosum seed, at least to a limited extent, constitutes a health hazard for certain animal species.

Toxicity of dietary Heliotropium dolosum seed to broiler chickens.

Five groups of 20 female broiler chicks were fed different levels of dehulled Heliotropium dolosum seed (w/w%; 0.0, 1.0, 3.0, 5.0 or 10.0%) from 10 to 52 d of age. In all doses the seed caused decreases in daily feed intake, weight gain, and feed efficiency, and biochemical findings, severity of pathologic changes, and mortality rate increased in a dose-dependent manner. Acute toxicity was observed in livers of chicks fed 10% seed. Other test groups had chronic changes. Livers had massive to submassive necrosis, hepatic megalocytosis, bile duct proliferation, fatty change, and periportal fibrosis. Biochemical evaluations revealed hypoalbuminemia, hypoprotienemia and increased ALP activity and billuribin. The seed of Heliotropium dolosum produced biochemical and specific pathologic changes in broiler chicks, as well as decreased food intake and feed efficiency. Higher seed levels induced more pronounced changes.