Zinc is a strong stimulant of metallothionein synthesis in the ischaemic testis tissue.

This study was performed to determine the effect of zinc supplementation effects on metallothionein levels in testis ischaemia-reperfusion of rats. The experimental groups were designed as Control, Sham, Ischaemia-Reperfusion (I/R) and I/R + Zinc supplemented. Zinc supplemented as 5 mg/kg day for 3 weeks. Testis tissues were analysed for metallothionein by immunohistochemical staining procedures. Group comparison showed that the zinc-supplemented ischaemia-reperfusion group had a significantly higher level of cells strongly stained with metallothionein than all other groups. A general evaluation of the results suggests that zinc supplementation is a strong stimulant of metallothionein synthesis in the ischaemic testis tissue.

Effect of pinealectomy and melatonin supplementation on metallothionein, ZnT2, ZIP2, ZIP4 and zinc levels in rat small intestine.

We investigated the relations among levels of metallothionein (MT); zinc (Zn) transport proteins, ZnT2, ZIP2 (ZRT and IRT-like proteins); and ZIP4, which enable Zn absorption in the small intestine of rats. We also investigated tissue Zn levels in the small intestine. We used four groups of adult male rats: group 1, control; group 2, pinealectomy (Px); group 3, Px + melatonin (MEL); group 4, MEL only. Animals in groups 3 and 4 were administered 5 mg/kg/day MEL for four weeks. At the end of the study, all animals were sacrificed and samples of duodenum, jejunum and ileum were harvested to analyze ZnT2, ZIP2, ZIP4 and MT levels using immunohistochemistry, and tissue Zn levels were measured by atomic absorption spectrophotometry. The lowest ZnT2 levels in the duodenum, jejunum and ileum, and the lowest ZIP2 levels in the duodenum and ileum were found in group 2. The lowest ZIP4 levels were found in the duodenum and jejunum, and the lowest MT levels in the duodenum and ileum were found in group 2. The highest MT values in the ileum were found in group 4. We found that ZnT2, ZIP2, ZIP4 and MT levels were reduced in the ileum compared to controls following Px, but levels approached control values after MEL administration. By its effects on ZnT2, ZIP2, ZIP4 and MT levels, MEL participates in the absorption of Zn in the rat small intestine.

Evaluation of the effects of vitamins C and E on experimental orthodontic tooth movement.

Background. This experimental study aimed to assess the effects of Vitamins C and E on orthodontic tooth movement. Methods. Fifty-one male Wistar albino rats were divided into six groups: five appliance groups and one control group. The appliance groups had an orthodontic appliance consisting of a closed-coil spring ligated between the maxillary incisor and maxillary first molar (50 g). Vitamin E and C (150 mg/kg) were injected intraperitoneally per day in the first and second groups, respectively. Vitamins E and C (20 µL) were locally injected into the periodontal gap of the moving teeth in the third and fourth groups, respectively, once every three days. No vitamin was injected in the last (fifth) appliance group.The experimental period was 18 days. Histological and biochemical (alkaline phosphatase, osteocalcin, and NTx levels) evaluations of the samples were performed, and maxillary incisor‒molar distance was measured before and after the experiment. Results. The amount of tooth movement was similar in the appliance groups. All the vitamin groups showed significantly increased osteoblastic activity, while those treated with systemic vitamins exhibited significantly increased numbers of collagen fibers on the tension side compared to the appliance control group (P<0.05). Conclusion. Vitamin C and E supplements positively affected bone formation on the tension side of the teeth during experimental orthodontic tooth movement.

Effects of zinc supplementation on metallothionein levels in ischemic renal tissue.

We investigated how zinc (Zn) supplementation affects metallothionein levels in the cortex and medulla of ischemic renal tissue of rats. We used adult male rats divided into four groups: group 1, untreated control; group 2, sham-operated; group 3, ischemia-reperfusion; group 4, ischemia-reperfusion + 5 g/kg Zn. Renal tissue was analyzed using immunostaining of rat metallothionein. Cells stained with metallothionein were counted and their percentage was calculated. We found that the Zn supplemented ischemia and reperfusion group exhibited a greater percentage of cells stained strongly for metallothionein in the renal cortex than all other groups. In the renal medulla, percentages of weak staining for metallothionein in the control and ischemia and reperfusion groups were greater than those in the sham and Zn-supplemented ischemia/reperfusion groups. Our findings indicate that the main effect of Zn in the renal tissue occurs in the cortex, while metallothionein synthesis in the renal medulla is unaffected.

Effect of Zinc and Melatonin on Oxidative Stress and Serum Inhibin-B Levels in a Rat Testicular Torsion-Detorsion Model.

The present study was aimed to examine the effects of 3-week zinc and melatonin administration on testicular tissue injury and serum Inhibin-B levels caused by unilateral testicular torsion-detorsion in rats. The study was performed on 60 Wistar Albino-type adult male rats. The animals were allocated to 6 groups in equal numbers. 1. Control; 2. Sham; 3. Ischemia-reperfusion; 4. Zinc + ischemia-reperfusion; 5. Melatonin + ischemia-reperfusion; 6. Zinc + melatonin + ischemia-reperfusion. Zinc and melatonin were administered before ischemia-reperfusion at doses of 5 and 3 mg/kg respectively, by intraperitoneal route for a period of 3 weeks. Testicular torsion-detorsion procedures consisted of ischemia for 1 h and then reperfusion for another hour of the left testis. Blood and testicular tissue samples were collected to analyze erythrocyte and tissue GSH and plasma and tissue MDA, Inhibin-B levels. The highest erythrocyte and testis GSH values were found in zinc, melatonin, and zinc + melatonin groups (p < 0.001). Torsion-detorsion group has significantly lower erythrocyte GSH levels and higher plasma MDA values (p < 0.001). Serum inhibin-B and spermatogenic activity levels in the torsion-detorsion group were also significantly lower than those in the other groups (p < 0.001). However, zinc-, melatonin-, and melatonin + zinc-supplemented groups have higher inhibin-B and spermatogenetic activity (p < 0.001). The results of the study show that zinc, melatonin, and melatonin + zinc administration partially restores the increased oxidative stress, as well as the reduced inhibin-B and spermatogenic activity levels in testes ischemia-reperfusion in rats. Suppressed inhibin-B levels in the testicular tissue may be a marker of oxidative stress.

The effect of dosage on the efficiency of LLLT in new bone formation at the expanded suture in rats.

The aim of this study was to investigate the effect that dosage has on the efficiency of low-level laser therapy (LLLT) in bone formation in a rat study model. Twenty-eight rats were divided into four groups as only expansion (OE), expansion + low dose (0.15 J) (LD), expansion + medium dose (0.65 J) (MD), and expansion + high dose (198 J) (HD) laser therapy groups. The midpalatal suture was expanded during 5 days. Afterwards, irradiations were started and performed with an 820 nm, continuous wave, Ga-Al-As diode laser (Doris, CTL-1106MX, Warsaw, Poland). At the end of experiment, the premaxillae of the animals were dissected. The sections were transferred into PC environment and analyzed by using Image Analysis program. Number of osteoblasts, osteoclasts, fibroblasts, vessels, transforming growth factor beta (TGF-ß) expression, and new bone formation were evaluated with this program. Amount of expansion did not show any difference among the groups. All parameters except the number of osteoclasts were increased in all lased groups while that parameter was significantly decreased. Vessels, TGF-ß expression, and new bone formation were mostly increased in LD group followed by HD group. Among the lased groups, a significant difference was observed only for the amount of new bone formation, which was between the LD and the MD groups. On the other hand, the difference in this parameter was insignificant between OE and MD groups. Low-level laser therapy with both 5 and 6,300 J/cm(2) doses was found to be significantly effective, while the 20 J/cm(2) dose did not show a significant effect in increasing new bone formation. This finding reveals that the efficiency of the therapy is affected by the dosage.

Interactive effects of melatonin, exercise and diabetes on liver glycogen levels.

BACKGROUND: This study aimed to examine the effects of melatonin supplementation on liver glycogen levels in rats with streptozotocin- induced diabetes and subjected to acute swimming exercise. MATERIAL AND METHODS: Eighty Sprague-Dawley type adult male rats were divided into eight groups: Group 1, general control; Group 2, melatonin-supplemented control; Group 3, melatonin-supplemented diabetes; Group 4, swimming control; Group 5, melatonin-supplemented swimming; Group 6, melatonin-supplemented diabetic swimming; Group 7, diabetic swimming; Group 8, diabetic control. Melatonin was supplemented at a dose of 3 mg/kg/day intraperitoneally for four weeks. Liver tissue samples were collected and evaluated using a Nikon Eclipse E400 light microscope. All images obtained from the light microscope were transferred to PC medium and evaluated using Clemex PE 3.5 image analysis software. RESULTS: The lowest liver glycogen levels in the study were found in group 4. Liver glycogen levels in groups 3, 6, 7 and 8 (the diabetic groups) were higher than group 4, but lower than those in groups 1 and 2. The lowest liver glycogen levels were obtained in groups 1 and 2. CONCLUSIONS: The study indicates that melatonin supplementation maintains the liver glycogen levels that decrease in acute swimming exercise, while induced diabetes prevents this maintenance effect in rats.

Effect of intraperitoneal selenium administration on liver glycogen levels in rats subjected to acute forced swimming.

There are a few of studies examining how selenium, which is known to reduce oxidative damage in exercise, influences glucose metabolism and exhaustion in physical activity. The present study aims to examine how selenium administration affects liver glycogen levels in rats subjected to acute swimming exercise. The study included 32 Sprague-Dawley type male rats, which were equally allocated to four groups: Group 1, general control; Group 2; selenium-supplemented control (6 mg/kg/day sodium selenite); Group 3, swimming control; Group 4, selenium-supplemented swimming (6 mg/kg/day sodium selenite). Liver tissue samples collected from the animals at the end of the study were fixed in 95% ethyl alcohol. From the tissue samples buried into paraffin, 5-µm cross-sections were obtained using a microtome, put on a microscope slide, and stained with PAS. Stained preparations were assessed using a Nikon Eclipse E400 light microscope. All images obtained with the light microscope were transferred to a PC and evaluated using Clemex PE 3.5 image analysis software. The highest liver glycogen levels were found in groups 1 and 2 (p < 0.05). The levels in group 4 were lower than those in groups 1 and 2 but higher than the levels in group 3 (p < 0.05). The lowest liver glycogen levels were obtained in group 3 (p < 0.05). Results of the study indicate that liver glycogen levels that decrease in acute swimming exercise can be restored by selenium administration. It can be argued that physiological doses of selenium administration can contribute to performance.

Haemostatic role and histopathological effects of a new haemostatic agent in a rat bladder haemorrhage model: an experimental trial.

OBJECTIVE: To investigate the haemostatic efficacy and histopathological effects of a new haemostatic agent, Ankaferd BloodStopper (ABS; Ankaferd Drug Cosmetic Co., Istanbul, Turkey) in a rat bladder haemorrhage model. ABS is a unique combination of five plant extracts that has been used in Turkish traditional medicine as a haemostatic agent for external traumatic bleeds. MATERIALS AND METHODS: In all, 20 male Sprague-Dawley rats were divided into two equal groups. In both groups, the mucosa was damaged on the posterior wall (PW) of the bladder. The liquid form of ABS was applied to the bleeding area of one group (group 1) and 0.9% NaCl to the bleeding area of the other group (group 2, controls). The solutions were applied drop by drop with a 2 mL injector until the bleeding stopped and the bleeding times recorded. For histopathological examination, two tissue samples were taken from all rats in each group; one from the damaged mucosa in the PW and one from undamaged mucosa in the lateral wall (LW). Two sections were prepared from all samples. One section was stained with haematoxylin and eosin (H&E), and the rest was used for immunohistochemical staining for fibronectin. RESULTS: The mean bleeding times were 65 s and 147 s for groups 1 and 2, respectively (P < 0.001). In the pathological specimens stained with H&E, fibrosis and other studied pathological features were similar in the LW samples in groups 1 and 2. Similarly, there were no differences between the PW samples of groups 1 and 2. In the pathological specimens stained for fibronectin, the number of attenuated cells was similar in the LWs of group 1 and 2 (P = 0.21) as well as in the PWs of each group (P = 0.066). CONCLUSIONS: ABS decreased bleeding time and did not increase fibrotic reactions in rat bladder tissue.

Efficacy of antioxidant treatment in the prevention of testicular atrophy in experimental testicular torsion.

BACKGROUND/PURPOSE: This study aimed to compare dose schedules of the antioxidant treatment (melatonin and steroid) used as 1 dose and as once a day for 7 days in terms of salvage of the testes in the late period. METHODS: Sixty prepubertal rats were divided into 6 groups each containing 10 rats: sham (S), torsion-detorsion (TD), 1-dose melatonin (M(1)), 1-dose steroid (ST(1)), 7-dose melatonin (M(7)), and 7-dose steroid (ST(7)) groups. The left testes were rotated 720 degrees for 6 hours and detorsed for 6 hours thereafter. In the treatment groups, 17 mg/kg melatonin and 1 mg/kg steroid were injected 15 minutes before detorsion. Left orchiectomies were performed to determine testicular weights and Johnsen scores 3 months later. RESULTS: Testicular weights and Johnsen scores in the M(1), ST(1), and TD groups were significantly lower compared with those in the S group, and atrophy developed in these groups, whereas they were higher in the M(7) and ST(7) groups compared with the TD group. Testicular atrophy did not develop in the M(7) or ST(7) groups. CONCLUSIONS: Our results suggested that antioxidant agents used once a day for 7 days prevent testicular atrophy and are effective in terms of salvage of the testes.

Effects of melatonin on spermatogenesis and testicular ischemia-reperfusion injury after unilateral testicular torsion-detorsion.

PURPOSE: This study aimed to determine the effect of melatonin in preventing ischemia-reperfusion (I-R) injury-induced tissue damage and on spermatogenesis after experimental testicular torsion (TT). METHODS: Forty peripubertal rats were divided into 4 groups each containing 10 rats: control (C), sham (S), torsion plus detorsion (TD), and torsion plus melatonin (M). The left testes were rotated 720 degrees for 6 hours and detorsed for 6 hours thereafter. Serum inhibin B (IB) levels were measured in blood samples taken from all groups. Left orchiectomies were performed to determine the tissue levels of Johnsen's scores (JS) and malondialdehyde (MDA). RESULTS: Serum IB levels in the S and TD groups were significantly lower compared with that in the C group, whereas they were higher in the M group compared with the TD group. The MDA levels were significantly lower in the C, S, and, M groups compared with the TD group. Johnsen's scores were significantly higher in the C, S, and M groups compared with the TD group. CONCLUSIONS: Our results suggested that melatonin is a potent antioxidant agent in preventing testicular I-R injury, as shown by increased IB levels and JS.

Effect of melatonin and zinc on the immune response in experimental Toxoplasma retinochoroiditis.

OBJECTIVES: To investigate the possible effect of melatonin (MEL) and zinc on the immune response to Toxoplasma gondii retinochoroiditis in the rat model of infection and to establish the possible value of artificial MEL and/or zinc supplementation as adjunctive therapeutic agents in the treatment of T. gondii retinochoroiditis. METHODS: Eighty-four Sprague-Dawley male rats were divided into 12 equal groups. All groups, except controls were infected with T. gondii parasite by intraperitoneal injection. Combinations of zinc-deficient diet, pinealectomy (Px), and artificial zinc and MEL were supplied during a 1-month period. At the end of the experiment, retinal and choroidal total lymphocytes, CD3+, CD4+, and CD8+ cell numbers were counted in histological sections. RESULTS: The highest amount of cellular infiltration (lymphocytes, CD3+, CD4+, CD8+ cells) in the choroid and retina was detected in infected + MEL + zinc-treated rats, and the least amount of cellular infiltration was observed in Px + zinc-deficient diet-treated rats. Although single zinc or MEL supplementation had no significant impact on the cellular infiltration in the retina and choroid in Px rats, combined therapy significantly improved these responses. CONCLUSION: Artificial supplementation of MEL and zinc should be considered as an adjunctive therapy to classic treatment of Toxoplasma retinochoroiditis especially in immunosuppressed and elderly patients if our data are confirmed in a clinical setting.

Biocompatibility of vitallium as ossicular reconstruction material in the middle ear: experimental animal study.

CONCLUSION: Although long-term data will be necessary for confirmation, the result of this preliminary study indicates that vitallium may be a good alternative material for ossicular replacement prostheses in the middle ear. OBJECTIVES: To investigate the biocompatibility of vitallium (Co-Cr-Mo) as ossicular reconstruction material in the rabbit middle ear, and to compare the results with those obtained with titanium, well known as a highly biocompatible material, and non-implanted control groups. MATERIAL AND METHODS: Eighteen female New Zealand White rabbits were anesthetized. The tympanomeatal flap was elevated and 12 vitallium and 12 titanium implants were placed in the bulla away from the ossicles in 24 middle ears. Six rabbits were used as non-implanted controls. All animals were sacrificed under general anesthesia on the 180th day after implantation. The temporal bones were removed, fixed in 10% buffered paraformaldehyde and decalcified for a week in EDTA. Tissue samples were then prepared using an Autotechnicon and embedded in paraffin. Sections (30-microm thick) were cut with a microtome, stained with hematoxylin-eosin, von Gieson's stain and fibroblast growth factor (FGF) and examined under a light microscope. The numbers of lymphocytes, collagen fibers and FGF-positive cells were determined in all three groups. RESULTS: There was no significant difference in the numbers of collagen fibers between the groups (p > 0.05). No significant differences were found in the numbers of lymphocytes and FGF-positive cells between the titanium and vitallium groups (p > 0.05). The differences in the numbers of lymphocytes and FGF-positive cells between the control and other groups were found to be significant (p <0.05).

The comparison of efficacies of topical corticosteroids and nonsteroidal anti-inflammatory drops on dry eye patients: a clinical and immunocytochemical study.

PURPOSE: To investigate whether conjunctival inflammation represents a primary event in the pathogenesis of keratoconjunctivitis sicca or whether it is a secondary inflammatory reaction caused by enhanced mechanical irritation as a result of surface dryness and whether anti-inflammatory drops (corticosteroids and nonsteroidal anti-inflammatory) have therapeutic effects and are similar. DESIGN: Single-masked, randomized, prospective clinical trial. METHODS: Thirty-two keratoconjuctivitis patients with or without Sjögren syndrome were included in the study. The patients were randomized to three groups. Group 1 patients received a topical artificial tear substitute (ATS); group 2 received ATS plus nonsteroidal anti-inflammatory drops (NSAID); and group 3 received ATS plus topical corticosteroidal drops. The eye symptom severity scores, Schirmer test values, rose bengal and fluorescein staining scores were evaluated before treatment and 15 and 30 days after start of treatment. Impression cytology specimens were stained using immunohistochemical methods to detect the percentages of human leukocyte antigen II (HLA-DR) positive, Apo 2.7 positive, and periodic acid-Schiff positive cells. Statistical analyses were performed within and between groups. Group 3 patients had significantly lower symptom severity scores, fluorescein and rose bengal staining, and HLA-DR positive cells on days 15 and 30 compared with patients in other groups. They also had a significantly higher number of periodic acid-Schiff positive (goblet) cells in their impression cytology specimens on days 15 and 30 compared with the other patients. On day 30, group 3 patients had significant differences compared with their baseline measurements in terms of above-mentioned parameters. However, we did not detect a significant effect of any treatment schedule on the Shirmer test value and the numbers of Apo 2.7 cells in impression cytology specimens. CONCLUSION: Topical corticosteroids had a clearly beneficial effect both on the subjective and objective clinical parameters of moderate-to-severe dry eye patients. These effects were associated with the reduction of inflammation markers of conjunctival epithelial cells.