The histological and biochemical analysis of the effects of radiofrequency radiation on testis tissue of rats and the protective effect of melatonin.

Background/aim: Primarily due to wireless communication devices, especially mobile phones, there has been a steady rise in the intensity of nonionizing radiofrequency radiation (RFR). In recent years, increased human health problems raised concerns about whether there is a positive relationship between intense exposure to RFR and public health. The present study aims to investigate the effects of GSM-like RFR exposure on the male reproductive system and the impact of melatonin treatment (synergistic, antagonist, or additive). Materials and methods: Thirty-six male Wistar Albino rats were used and separated into six groups: i. Control; ii. Sham; iii. RFR exposure; iv. Control-melatonin; v. Sham-melatonin; vi. Melatonin + RFR exposure. Animals were exposed to 2600 MHz RFR with electric (E) field levels of 21.74 V/m for 30 min per day, 5 days per week, for 4 weeks. All testicular tissue samples were evaluated under a light microscope for hematoxylin-eosin staining. Biochemical analyses were performed by measuring malondialdehyde, total nitric oxide, glutathione, and glutathione peroxidase levels. We evaluated the combined effects of prolonged RFR exposure and melatonin treatment on ROS-mediated structural changes in testicular tissues. Results: Results showed that reactive intermediates (malondialdehyde and total nitric oxide) increased significantly with RFR exposure, while the protective effect of melatonin effectively reduced the radical levels of the tissues. Histological evaluation revealed a decrease in cell population and connective tissue elements under RFR exposure, accompanied by marked edema in the testicular tissues. Conclusion: The structural and functional effects of prolonged RFR exposure might be ROS-based. Moreover, these adverse effects might be compensated with externally treated supplements. There is a need for new extensive research.

Therapeutic effects of melatonin in long-term exposure to 2100MHz radiofrequency radiation on rat sperm characteristics.

INTRODUCTION: Radiofrequency electromagnetic fields (RF-EMFs) are one of the risk factors for male reproductive health and melatonin can be an ideal candidate for therapeutic development against RF-induced male fertility problems due to its antioxidant properties. The possible therapeutic role of melatonin in the destructive effects of 2100MHz RF radiation on rat sperm characteristics is investigated in the present study. METHODS: Wistar albino rats were divided into four groups and the experiment continued for ninety consecutive days; Control, Melatonin (10mg/kg, subcutaneously), RF (2100MHz, thirty minutes per day, whole-body), and RF+Melatonin groups. Left caudal epididymis and ductus deferens tissues were placed in sperm wash solution (at 37°C) and dissected. The sperms were counted and stained. Measurements of the perinuclear ring of the manchette and posterior portion of the nucleus (ARC) were performed and the sperms were examined at an ultrastructural level. All of the parameters were evaluated statistically. RESULTS: The percentages of abnormal sperm morphology were significantly increased with RF exposure, while the total sperm count was significantly decreased. RF exposure also showed harmful effects on acrosome, axoneme, mitochondrial sheath, and outer dense fibers at the ultrastructural level. The number of total sperms, sperms with normal morphology increased, and ultrastructural appearance returned to normal by melatonin administration. DISCUSSION: The data showed that melatonin may be a beneficial therapeutic agent for long-term exposure of 2100MHz RF radiation-related reproductive impairments.

Melatonin effective to reduce the microscopic symptoms of polycystic ovary syndrome-related infertility: An experimental study.

Polycystic ovary syndrome (PCOS) is an endocrine disorder seen in women of reproductive age and has been gradually increasing over the years. The mechanism of the syndrome has still not been clearly understood. In this study, the possible effects of exogenously administrated melatonin on melatonin (MT1) receptor, Growth Differentiation Factor-9 (GDF9), and Bone Morphogenetic Protein-15 (BMP15) in experimental PCOS were investigated. Thirty-two 6-8-week-old Sprague-Dawley rats were randomly divided into four groups (n = 8 in each) as Sham control (Group 1), Melatonin (Group 2), PCOS (Group 3), and PCOS + Melatonin (Group 4) groups. At the end of the 21st day, the experiment was terminated, the ovary tissues were taken, and Hematoxylin-Eosin staining, MT1, GDF9, BMP15 immunohistochemical labeling, western blot, and quantitative real-time polymerase chain reaction (qPCR) analyses were performed. Serum Luteinizing Hormone (LH)/Follicle Stimulating Hormone (FSH) levels and colpo-cytological examinations were also carried out. The results revealed that melatonin administration increased the expression levels of the MT1 receptor, GDF9, and BMP15 in PCOS at protein and mRNA levels. It was determined that melatonin administration reduced the microscopic symptoms of PCOS. Melatonin was found to be effective via the MT1 receptor in the pathogenesis of PCOS, and it suppressed the transport pathways of GDF9 to granulosa cells in antral follicles.

The effect of antiepileptic drugs on re-myelinization of axons: Phenytoin, levetiracetam, carbamazepine, and valproic acid, used following traumatic brain injury.

OBJECTIVE: Traumatic brain injury is a major health and socioeconomic problem and the first cause of young death worldwide. For this reason, the prevention of post-traumatic brain injury and the research of new methods for it are important today. In this study, we aimed to determine whether the use of antiepileptic drugs contributed to axonal healing after traumatic brain injury. METHODS: Thirty-six Long-Evans rats, each weighing 300-350 g, were used in this study. A total of 6 groups, including the sham, control, and 4 study groups, were determined. A 1.5 mm-sized trauma was created in the biparietal area with a blunt-tipped dissector. Carbamazepine phenytoin valproic acid and levetiracetam (phenytoin: 30 mg/kg, valproic acid: 60 mg/kg, levetiracetam: 80 mg/kg, and carbamazepine: 36 mg/kg) were intraperitoneally administered to the study groups, and the control group intraperitoneally received a physiological saline solution (15 ml/kg) twice daily for 3 days. After 72 h, hemispheres of the sacrificed subjects were taken for examination in biochemistry and histology. Glutathione, malondialdehyde, and NG2 levels in the samples were determined. RESULTS: No significant difference was found in biochemical measurements. Histopathological examination revealed that the NG2 expression was more intense in the group treated with phenytoin and levetiracetam (phenytoin was partly higher) and the amount of edema decreased. The NG2 expression increased and the edema decreased, though lower in the group treated with carbamazepine and valproic acid, compared with phenytoin and levetiracetam. An increase in the NG2 expression and edema intensity were determined in the control and sham groups. CONCLUSION: Antiepileptic drug selection after traumatic brain injury is an important medical matter. Although the patient-oriented selection is essential, the study suggests that the choice of phenytoin, levetiracetam carbamazepine, and valproic acid will, respectively, have an accelerating effect for axonal healing.

Involvement of endometrial IGF-1R/IGF-1/Bcl-2 pathways in experimental polycystic ovary syndrome: Identification of the regulatory effect of melatonin.

The involvement of endometrial IGF-1R/IGF-1/Bcl-2 pathways and the potential regulatory effects of exogenously administrated melatonin on this expression is investigated in the experimental PCOS model in the present study. Thirty-two 6-8 week old Sprague Dawley rats were divided into four groups: the Sham Control Group (1% CMC/day by oral gavage [o.g.]); the Melatonin Group (2 mg/kg/day melatonin by subcutaneous administration [s.c.]); the Experimental PCOS Group (1 mg/kg/day Letrozole by o.g.); and the Experimental PCOS + Melatonin Group (1 mg/kg/day Letrozole by o.g. and 2 mg/kg/day melatonin by s.c. administration). Vaginal smear samples were taken from the 14th day to the end of the experiment for colpocytological measurements. At the end of the 21 day experimental period, uterine tissues were taken; Hematoxylin-Eosin histochemical, IGF-1R/IGF-1/Bcl-2, PCNA immuno-histochemical stainings and western blot analyses were performed for related antibodies. All of the data was supported statistically. The epithelium of endometrium lost its single-layer structure in some parts, separation was observed between the epithelium and the basal membrane junction, intracellular edema was found in the uterine glands by the polycystic ovary-induction. Also this induction increased the expression of IGF-1R/IGF-1, Bcl-2, and PCNA proteins. Morphological degenerations returned to its normal appearance generally by the melatonin administrations and melatonin also regulated the increased expression of endometrial IGF-1R/IGF-1/Bcl-2 and PCNA pathways. It is concluded that additional studies are needed, using melatonin as a supporting agent may be appropriate in cases of PCOS.

Arterial Occlusion After Hyaluronic Acid Injection: Treatment With Hyaluronidase and Streptokinase.

ABSTRACT: The most feared complication of the hyaluronic acid injections in the periorbital region is embolism of the central retinal artery. The present study aimed to compare the effectiveness of hyaluronidase administered intravenously (systemically) alone or in combination with streptokinase with that of intra-arterial revascularization. Thirty rats were divided into 5 groups. The bilateral oblique groin flap of the rats was raised; the right side was the experiment group, and the left side was the sham control. The right superficial epigastric artery was occluded with a hyaluronic acid injection. After occlusion, no additional procedures were performed in group 1, whereas group 2 received systemic hyaluronidase, group 3 received intra-arterial hyaluronidase, group 4 received systemic hyaluronidase and streptokinase, and group 5 received intra-arterial hyaluronidase and streptokinase. On the seventh day, the rats were killed, flap necrosis rate was calculated, and histological examination was performed. There was no significant difference in the necrosis rates of the rats in groups 2, 3, 4, and 5 (P > 0.05). In histological evaluation, the histological view closest to normal arterial structure was observed in group 4. Immunohistochemical analysis revealed that the ischemia scores of systemic therapy were significantly lower than those of intra-arterial therapy. These results have shown that hyaluronidase and streptokinase administered systemically is as effective as intra-arterial revascularization and does not cause arterial wall degeneration. It has been shown that systemic administration of hyaluronidase and streptokinase is as successful as intra-arterial revascularization in the treatment of arterial embolism with hyaluronic acid.

Melatonin Modulates NMDA-Receptor 2B/Calpain-1/ Caspase-12 Pathways in Rat Brain After Long Time Exposure to GSM Radiation.

AIM: To investigate the potential protective effects of melatonin on the chronic radiation emitted by third generation mobile phones on the brain. MATERIAL AND METHODS: A total of 24 male Wistar albino rats were divided into four equal groups. Throughout a 90-day experiment, no application was performed on the control group. The second group was exposed to 2100 MHz radiation for 30 minutes. Subcutaneous melatonin was injected into the third group. Subcutaneous melatonin injection was applied 40 minutes before radiation and then the fourth group was exposed to radiation for 30 minutes. At the end of the experiment, brain (cerebrum and cerebellum) tissues were taken from the subjects. Histochemical, immunohistochemical, ultrastructural and western blot analyses were applied. In addition to brain weight, Purkinje cells’ number, immunohistochemical H Score analyses and the results of the Western blot were examined statistically. RESULTS: With the application of radiation, neuronal edema, relatively-decreased numbers of neurons on hippocampal CA1 and CA3 regions, displacement of the Purkinje neurons and dark neurons findings were observed as a result of histochemical stainings. Radiation also activated the NMDA-receptor 2B/Calpain-1/Caspase-12 pathway, NMDA-receptor 2B and Calpain-1 with the findings being supported by western blot analyses. Pre-increased protein synthesis before apoptosis was identified by electron microscopy. CONCLUSION: Mobile phone radiation caused certain (ultra) structural changes on the brain and activated the NMDA-receptor 2B/ Calpain-1/Caspase-12 pathway; in addition, melatonin was found to be effective, but insufficient in demonstrating the protective effects.

Penetration Depth in Nanoparticles Incorporated Radiofrequency Hyperthermia into the Tissue: Comprehensive Study with Histology and Pathology Observations.

In present study, the effective penetration of radiofrequency (RF) induced gold decorated iron oxide nanoparticles (GS@IONPs) hyperthermia was investigated. The effective penetration depth of RF also the damage potency of hyperthermia was evaluated during histopathology observations which were done on the chicken breast tissue and hepatocellular carcinoma (HCC) models. The thermal damages are well- documented in our previous cellular study which was engaged with potency of RF hyperthermia in Epithelial adenocarcinoma (MCF-7) and fibroblast (L-929) cells deaths [1]. In recent work, PEGylated iron oxide nanoparticles (IONPs) were used as base platform for gold magnetic nanoparticles (GS@IONPs) formation. The 144.00015 MHz, 180W RF generator was applied for stimulating the nanoparticles. The chicken breast tissue and the hepatocellular tumor model was considered in the experimental section. In histology studies, the structural changes also the effective penetration depth of RF induced nanoparticles was observed through microscopic monitoring of the tissue slices in histology observations (Gazi medical school). The highest damage level was seen in 8.0 µm tissue slices where lower damages were seen in depth of 1.0 cm and more inside tissue. The histology observations clarified the effective penetration depth of RF waves and irreversible damages in the 2.0 cm inside the tissue.

Is leptin receptor expression triggered in the case of embryo transfer to endometrium coculture?

Background/aim: A synchronized dialogue between maternal and embryonic tissues is required for successful implantation. Low uterine receptivity is responsible for two-thirds of implantation failures and leptin is effective in the physiology of reproduction by binding to specific receptors. In this study, we investigateleptin receptor expression in cases of embryo transfer to endometrial coculture. Materials and methods: Biopsy materials were taken from 20 females with indication for coculture application and were cultured in an appropriate medium after the epithelial cells were isolated. The grown cells were cultured in chamber slides as the first group. For the second group, day 3 embryo was added to chamber slides and the development was observed. The embryo was transferred 1 or 2 days later and other cells (after the transfer process) were used to form the second group. After fixation, immunohistochemical staining was performed with anti-leptin primary antibody. Results: Regarding the coculture without embryo transfer, moderate leptin receptor immunoreactivity was seen in the perinuclear region and the cell membrane. Also, regarding the coculture with embryo transfer, moderate leptin receptor immunoreactivity was seen in the cytoplasm and strong leptin receptor immunoreactivity was seen in the cell membrane. Conclusion: Embryo transfer to endometrium coculture triggers leptin receptor expression

Sutureless approach with vein grafts and mesenchymal stem cells in primary nerve repair: Functional and immunohistological results.

PURPOSE: The aim of this study was to define a sutureless peripheral nerve repair technique with a vein graft and bone marrow-derived stem cells (BMSC) and compare it to epineural repair. MATERIALS AND METHODS: Thirty Wistar Albino rats were divided into five groups evenly. In the control group (C), epineural repair was performed. In the SV (suture + vein) and MSV (BMSC + suture + vein) groups, epineural repair was wrapped with a vein graft. In the V (vein) and MV (vein + BMSC) groups, sutureless repair using a vein graft was performed by taking sutures away from the regeneration site. Rats were evaluated with pinprick, toe spread tests and sciatic nerve index (SFI) at 4th, 8th, and 12th weeks. They were sacrificed at 12th week, repair sites were harvested and evaluated immunohistochemically. RESULTS: There was no difference in pinprick and toe spread tests between the groups at 12th week. The mean SFI was -76.5 ± 3.7, -65.2 ± 11.7, -46.2 ± 19.4, -68.8 ± 9.8, -56 ± 8.8 in the C, SV, MSV, V, MV groups, respectively. The MSV group showed significantly the best SFI results (P < .05). NF-H immunostaining scores were as C; 1 ± 0.18, SV; 2.5 ± 0.36, MSV; 4 ± 0.49, V; 1.56 ± 0.54, MV; 3 ± 0.39, whereas GAP-43 scores were as C; 1 ± 0.31, SV; 2.66 ± 0.56, MSV; 4.50 ± 0.23, V; 2 ± 0.23, MV; 3 ± 0.6. The best nerve regeneration according to immunostaining results was observed in the MSV group (P < .05). The mean fibrosis area was 221.5 ± 25.9, 101.6 ± 7.1, 121.3 ± 18.8, 150.3 ± 12.1, 152.4 ± 11.8 µm2 in the above groups, respectively. SV and MSV groups showed the significantly less fibrosis area (P < .05). CONCLUSION: Epineural suture repair combined with vein wrapping and BMSCs (MSV) showed the best SFI, GAP-43, and NF-H immunostaining results.

Botulinum Toxin Type A: Assessing The Effects on The Brain Stem.

BACKGROUND: In this study, our aim is to investigate the possible effects of Botulinum toxin type A administrations in the early and late periods on the brain stem. METHODS: Eighteen white New Zealand rabbits were used in this study with the subjects being divided into three groups. Group I received 0.05 mL sterile saline to the left anterior auricular muscles. Group II and III were injected with Botulinum toxin type A (Botox, Allergan) to the left anterior auricular muscles. Group II was sacrificed 5 days after application and Group III was sacrificed 12 weeks after application; brain stem tissues were then taken. The samples were examined with Caspase 3, 8, and 9 immunohistochemical stainings. RESULTS: In the control group with Caspase-3 immune staining, moderate-to-strong immune reactivity was seen in a small number of neurons. In the Caspase-8 and 9 immune stainings, the immune reactive neurons were seen in greater numbers when compared with the Caspase-3 immune reactive neurons. In the early and late period, groups with Caspase-8 and 9 immune stainings, the immune reactive neurons were seen in greater numbers and in the wider area when compared with the Caspase-3 immune reactive neurons. No significant differences were recognized in the Caspase immune stainings between the early and late period groups. The results were statistically supported. CONCLUSION: It was concluded that Botulinum toxin type A application did not trigger apoptosis in stem cell tissues. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Effects of Low-Level Laser Therapy With a Herbal Extract on Alveolar Bone Healing.

PURPOSE: This study investigated the effect of a gallium-aluminum-arsenide (GaAlAs) diode laser used in low-level laser therapy (LLLT) with the application of Mecsina Hemostopper on mandibular alveolar bone healing. MATERIALS AND METHODS: Standard semispherical bone defects were created in left mandibular diastema sites of 32 female Long-Evans rats. Experimental animals were allocated to 1 of 4 groups: control group (no treatment), laser group (GaAlAs LLLT), Mecsina group, and laser-Mecsina combination group. Liquid Mecsina 0.01 mL was applied to the bone defects. Laser treatment was performed for 7 days after surgery at an energy dose of 10 J/cm2. All animals were sacrificed to observe hard tissue healing histologically, immunohistochemically, and radiologically at 30 days after surgery. RESULTS: Histologic assessment showed significantly more calcified tissue areas and significantly more osteoblast cells in the laser and laser-Mecsina groups than in the other groups (P < .01). Qualitative morphologic assessment showed that more bone tissue was present in the laser-Mecsina group than in the other groups. CONCLUSION: This study showed that LLLT, Mecsina application, and combined treatments were effective in healing alveolar bone among all tested treatment modalities.

The Effect of Hyaluronic Acid Application on the Perisilicon Capsule Structure.

BACKGROUND: Silicon implants constitute a major part of plastic surgery practice. Although materials with high biocompatibility have been used around the implants, capsule formation still develops and progressive nature of this process results in capsule contraction. The aim of this study is to evaluate the effects of hyaluronic acid injected around the silicon block on the capsule structure. METHODS: Twenty Wistar albino rats were used in the study. Rats were divided into two main groups (group 1 and group 2) and two subgroups. Rats in group 1 were sacrificed in week 4 and rats in group 2 were sacrificed in week 8. A subcutaneous pouch was created in the dorsum of the rats and a silicon block was placed into the pouch in groups 1A and 2A. 0.2 ml of hyaluronic acid was injected around the silicon block in group 1B and group 2B. Rats were sacrificed and capsule structure and thickness were analyzed following macroscopic evaluation. Concentrations of transforming growth factor-ß1 (TGF-ß1) and heat shock protein-47 (HSP-47) were evaluated immunohistochemically, and statistical comparisons were made. RESULT: Capsule structure consisted of three layers in all the groups. A more intense collagen structure was observed in the middle layer. The capsule was thinnest in group 1A and thickest in group 2B; the difference between the groups was statistically significant. TGF-ß1 was most intense in group 2B and it was correlated with the amount of collagen. Involvement of HSP-47 was observed mainly in collagen and also in fibroblasts and vascular structures, and its concentration was found to be lower in groups 2A and 2B. CONCLUSION: Exogenously added cross-linked hyaluronic acid increased the capsular thickness and may increase the risk of developing capsular contracture around silicone implants. LEVEL OF EVIDENCE II: Evidence was obtained from the well-designed controlled trials without randomization.

Is there crosstalk between subchondral bone, cartilage, and meniscus in the pathogenesis of osteoarthritis?

OBJECTIVES: This study aims to investigate if there is any crosstalk between subchondral bone, cartilage, and meniscus in the pathogenesis of osteoarthritis. PATIENTS AND METHODS: Twelve female patients (mean age 64 years; range 59 to 71 years) with osteoarthritis in medial compartment were included in the study. The samples of subchondral bone, cartilage and meniscus were obtained during total knee arthroplasty. Degenerated tissue samples obtained from medial compartment were used as the experimental group (12 samples of subchondral bone and cartilage, 1x1 cm each; and 12 samples of meniscus, 1x1 cm each). Healthy tissue samples obtained from lateral compartment were used as the control group (12 samples of subchondral bone and cartilage; 1x1 cm each; and 12 samples of meniscus, 1x1 cm each). After decalcification, tissue samples were evaluated with light and transmission electron microscopy. RESULTS: In the experimental group, light microscopic evaluation of subchondral bone samples demonstrated that the cartilage-to-bone transition region had an irregular structure. Degenerated cartilage cells were observed in the transition region and bone cells were significantly corrupted. In the experimental group, light microscopic evaluation of the meniscus samples demonstrated that the intercellular tissue was partly corrupted. Separation and concentration of the collagen fibers were evident. All findings were supported with ultra structural evaluations. CONCLUSION: Our findings indicate that degeneration of subchondral bone, cartilage, and meniscus probably plays a role in the pathogenesis of osteoarthritis with crosstalk.