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Rheumatoid arthritis (RA) is a multifactorial autoimmune disease that affects the joints of approximately 1 % of the population worldwide and is seen 2-4 times less in males. INSL3 is a gender-specific peptide hormone produced much higher in males than in females and may have an anti-inflammatory role in RA. So, in this study, it was aimed to determine the possible anti-inflammatory effect and dose of insulin-like factor-3(INSL3) in an experimental Complete Freund's adjuvant(CFA)-induced RA male rat model and lipopolysaccharide(LPS)-induced macrophage cell line and compare it with prednisolone therapy. For in vivo experiments, 48 male mice were randomly divided into 6 groups with 8 subjects in each group: Control group, Arthritis group, Arthritis + Prednisolone(10 mg/kg) group, Arthritis + INSL3(0.08-0.8-8 μg/day) groups. Joint tissue samples obtained from sacrificed subjects were examined by histochemical and immunohistochemically methods after biometric analyses, arthritis severity scoring, and thermal latency experiments. LPS-induced macrophage cells were also treated with prednisolone(1 µg/ml) and INSL3(50-100-200 nM). Cell viability, cell morphology, and TNF-α and IL-6 immune reactivity were evaluated. According to the data obtained from in vivo analyses, it was seen that INSL3 reduced the paw diameter and arthritis severity scoring, degenerative changes, and inflammation and increased the thermal latency, compared to the arthritis group, although not as much as the prednisolone treatment group. In vitro analyses showed that high doses of INSL3 had positive effects on cell viability, morphology, TNF-α, and IL-6 immune reactivity. In conclusion, it was determined that the anti-inflammatory effect of INSL3 was not as pronounced as prednisolone, but it had a more favorable impact on macrophage cell viability and morphology. It was concluded that INSL3 may be a protective therapeutic agent in combination with existing treatment protocols in treating many autoimmune diseases., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Gulfidan Coskun reports financial support was provided by Cukurova University Faculty of Medicine. Gulfidan Coskun reports a relationship with Cukurova University Faculty of Medicine that includes: employment., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: Successful embryo implantation is contingent upon the intricate interaction between the endometrium and the blastocyst. Recurrent implantation failure (RIF) signifies the clinical challenge of failing pregnancy post-transfer of high-quality embryos, fresh or frozen, in at least three in vitro fertilization (IVF) cycles, often in women under 40 years. Recent studies identify impaired blastocyst maternal tissue communication among recurrent implantation failure causes. Despite successful embryo transfer in vitro fertilization cycles, endometrial factors persist in women with recurrent implantation failure history, underscoring implantation's complexity. The implantation window, during which the endometrium becomes receptive to the blastocyst, involves the expression of key molecules that facilitate the implantation process. When the literature was examined, it was observed that comparative immunohistochemical studies on key molecules such as α4β1 integrin, MUC-2 and PNAd, which are thought to play a critical role in the endometrium before and during implantation, were limited. In this study, we aimed to investigate, through immunohistochemical and histological analyses, the roles of adhesion molecules in the secretory phase endometrium of patients diagnosed with RIF., Design: Twenty-one patients diagnosed with recurrent implantation failure and 21 patients with a history of previous pregnancy at the Gynecology and Obstetrics Clinic of Balcalı Hospital, Faculty of Medicine, Çukurova University were clinically evaluated between days 19-21 of the menstrual cycle and included in the study. Endometrial biopsies, prepared for light and electron microscopy, received Hematoxylin and Eosin staining and anti-α4β1 integrin, anti-MUC-2, and anti-PNAd antibodies for immunohistochemistry. Blood samples were collected on the 2nd and 3rd days of the menstrual cycle to assess serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and thyroid-stimulating hormone (TSH)., Results: While FSH, LH, and E2 levels showed no significant difference, TSH was elevated in the recurrent implantation failure group. Structural differences in the endometrium included increased microvilli cells and reduced pinopod counts in infertile women compared to controls. In women with recurrent implantation failure MUC-2 expression were found to be elevated in the endometrial surface epithelium, while PNAd expression was reduced compared to the control group., Conclusion: These findings suggest structural and molecular disparities during the endometrial receptivity window in recurrent implantation failure women may underlie infertility by hindering blastocyst adherence. Molecular studies are needed to elucidate the pathophysiology of the biomarkers whose presence in the endometrium we examined immunohistochemically and to discover new molecules., Capsule: Changing expressions of adhesion molecules (PNAd, α4β1 and MUC-2) under the influence of steroid hormones, remodeling of intercellular connections and stromal epithelial communication play an important role in endometrial receptivity., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ali ASKIN reports financial support was provided by Mersin University. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Purpose: Reducing renal ischemia is crucial for the function and survival of grafts from nonheartbeat donors, as it leads to inflammatory responses and tubulointerstitial damage. The primary concern with organs from nonheartbeat donors is the long warm ischemia period and reperfusion injury following renal transplantation. This study had two main goals; one goal is to determine how Necrostatin-1 targeting the PANoptosome affects PANoptosis in the nonheart-beating donor rat model. The other goal is to find out if Necrostatin-1 can protect the kidney from ischemic injury for renal transplantation surgery., Methods: Twenty-four rats were grouped randomly as control and Necrostatin-1 in this experimental animal study, and we administered 1.65 mg/kg of Necrostatin-1 intraperitoneally to the experimental group for 30 minutes before cardiac arrest. We removed the rats' left kidneys and measured various oxidative stress marker measures such as malondialdehyde, superoxide dismutase, catalase, GPx, and 8-hydroxy-2-deoxyguanosine levels. We then subjected the tissues to immunohistochemical analysis, electron microscopy, and histopathological analysis., Findings: The Necrostatin-1 group had a lower total tubular injury score (P < .001) and less Caspase-3, gasdermin D, and mixed lineage kinase domain-like protein expression. Additionally, the apoptotic index of the study group was lower (P < .001). Furthermore, the study group had higher levels of superoxide dismutase and GPx (P < .05), whereas malondialdehyde levels were reduced (P = .009). Electron microscopy also revealed a significant improvement in tissue structure in the Necrostatin-1 group., Conclusion: Necrostatin-1 protects against ischemic acute kidney injury in nonheart-beating donor rats by inhibiting PANoptosis via the blockade of RIPK1. As a result of this, Necrostatin-1 may offer novel opportunities for protecting donor kidneys from renal ischemia-reperfusion injury during transplantation in patients with end-stage kidney disease requiring a renal transplantation., Competing Interests: Declaration of competing interest None declared., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Context: Inflammasome NLR family pyrin domain-containing 3 (NLRP3) is associated with neurological disorders. Neuroinflammation can be suppressed by inhibiting NLRP3 inflammasome activation, decreasing neurodegenerative disorder progression. We devised a therapeutic technique that can reduce neuroinflammation induced by microglial activation, avoiding neurodegeneration. We aimed to investigate the mechanisms underlying the pharmacological effects of galantamine and wedelolactone by evaluating the response of the nuclear factor kappa B (NF-κB) signaling pathway and NLRP3 inflammasome in lipopolysaccharide (LPS)-activated N9 microglia., Methods: LPS and adenosine triphosphate were used to activate the NLRP3 inflammasome in N9 microglial cells, which were pretreated with galantamine and wedelolactone. Caspase-1, NLRP3, NF-κB, and interleukin (IL)-1β levels were measured using RT-qPCR and immunostaining., Results: Combined administration of galantamine and wedelolactone rescued microglial cells from LPS-induced cell death. Furthermore, treatment with galantamine and wedelolactone led to the suppression of NF-κB expression. NLRP3, caspase-1, and IL-1β levels were decreased by the combined treatment., Discussion and Conclusion: The concurrent administration of galantamine and wedelolactone effectively suppresses the production of inflammatory cytokines and NLRP3 inflammasome activation in microglia. This inhibitory effect is likely linked to the NF-κB signaling pathway modulation. Therefore, this combined treatment is a potential therapeutic approach for neuroinflammatory diseases.

Background: This study aimed to investigate cell degeneration, apoptosis, and ultrastructural differences in refractive lenticules (RL) obtained using small incision lenticule extraction (SMILE) compared with spherical equivalence (SE) refraction values., Methods: This study included 84 eyes from 42 patients. Patients were divided into two groups according to the SE values: those with values below 4 diopters (D) (Group 1) and above 4 diopters (D) (Group 2). Patients who did not belong to the same SE group were excluded from the study. One RL obtained from each patient was separated for light microscopy and immunohistochemical examinations, and another for transmission electron microscopy (TEM) examinations. Caspase-3 for apoptosis and alpha-smooth muscle actin (α-SMA) for cell degeneration were evaluated using immunohistochemical examinations., Results: Histological analyses showed that the density of collagen fibres was greater in Group 1 than in Group 2. Glycoaminoglycan and glycoprotein staining intensities were also higher in Group 1. TEM observations showed that Group 1 had intact cell and nuclear membranes, peripheral heterochromatin, and large nuclei, while Group 2 showed heterochromatin condensation and fragmentation, increased intracellular vacuoles, and loss of cytoplasm. Immunohistochemical analyses revealed that α-SMA and caspase-3 were significantly higher in Group 2 than in Group 1 (p < 0.001 and p < 0.001, respectively)., Conclusions: Cell degeneration and apoptosis were significantly more common in the RLs with high SE values after SMILE surgery. The tissue response induced by surgery was more severe in the RLs with high SE values. This should be considered when reusing RLs., (© 2024 The Author(s). Clinical & Experimental Ophthalmology published by John Wiley & Sons Australia, Ltd on behalf of Royal Australian and New Zealand College of Ophthalmologists.)

Despite research, the role of exercise in treatment and prevention of neurodegenerative diseases remains unclear. Our study, investigated that protective effect of treadmill exercise on molecular pathways and cognitive behaviours in a scopolamine-induced model of Alzheimer's disease. For that purpose, male Balb/c mice subjected to exercise for 12 weeks. During the last 4 weeks of exercise, mice were given an injection of scopolamine (2 mg/kg). Following injection, open field test and Morris water maze test were used to assess emotional-cognitive behaviour. Hippocampus and prefrontal cortex of mice were isolated, and levels of BDNF, TrkB, and p-GSK3ß Ser389 were assessed by western blotting, and levels of APP and Aß-40 were analysed by immunohistochemistry. In our study, scopolamine administration increased anxiety-like behaviour in open field test, while negatively affecting spatial learning and memory in Morris water maze test. We found that exercise had a protective effect against cognitive and emotional decline. Scopolamine decreased levels of p-GSK3ß Ser389 , BDNF in hippocampus and prefrontal cortex.Whereas TrkB decreased in hippocampus and increased in prefrontal cortex. There was an increase in p-GSK3ß Ser389 , BDNF, TrkB in the hippocampus, and p-GSK3ß Ser389 , BDNF in the prefrontal cortex in the exercise + scopolamine group. Immunohistochemical analysis showed that scopolamine administration increased APP and Aß-40 in hippocampus and prefrontal cortex in neuronal and perineuronal areas whereas Aß-40 and APP were reduced in exercise + scopolamine groups. In conclusion, long-term exercise may have a protective effect against scopolamine-induced impairments in cognitive-emotional behaviour. It can be suggested that this protective effect is mediated by increased BDNF levels and GSK3ß Ser389 phosphorylation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objectives: The objectives of this study were a) to investigate the effect of targeting the PANoptosome with 3,4-methylenedioxy-β-nitrostyrene (MNS) on PANoptosis in the Renal ischemia-reperfussion (RIR) model b) to investigate the kidney protective effect of MNS toward RIR injury., Methods: Thirty-two rats were divided into four groups randomly. The groups were assigned as Control, Sham, DMSO (dimethyl sulfoxide) and MNS groups. The rats in the MNS group were intraperitoneally given 20 mg/kg of MNS 30 minutes before reperfusion. 2% DMSO solvent that dissolves MNS were given to the rats in DMSO group. Left nephrectomy was performed on the rats under anesthesia at the 6th hour after reperfusion. Glutathione peroxidase (GPx), malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) and 8-Okso-2'-deoksiguanozin (8-OHdG) levels were measured. Immunohistochemical analysis, electron microscopic and histological examinations were carried out in the tissues., Results: Total tubular injury score was lower in the MNS group ( p  < 0.001). Caspase-3, Gasdermin D and MLK (Mixed Lineage Kinase Domain Like Pseudokinase) expressions were considerably decreased in the MNS group ( p  < 0.001). Apoptotic index (AI) was found to be low in the MNS group ( p  < 0.001). CAT and SOD levels were higher in the MNS Group ( p  = 0.006, p  = 0.0004, respectively). GPx, MDA, and 8-OH-dG levels were similar ( p  > 0.05) in all groups. MNS considerably improved the tissue structure, based on the electron microscopic analysis., Conclusions: Our results suggested that MNS administrated before the reperfusion reduces pyroptosis, apoptosis and necroptosis. These findings suggest that MNS significantly protects the kidney against RIR injury by reducing PANoptosis as a result of specific inhibition of Nod-like receptor pyrin domain-containing 3 (NLRP 3), one of the PANoptosome proteins.

Background/aim: We have designed an adjustable bone plate (ABP) which allows bone shortening and lengthening after fixation, which is a property not present in any of the plate systems available today. The aim of the current study was to examine the new ABP's segmental bone transfer capability for the treatment of a segmental bone defect in an animal model., Materials and Methods: Five sheep had ABPs attached to 10 of their tibias and bone defects of 15 mm in size were created. The pinion mechanism was moved with a manual screwdriver at a rate of 1mm/day for 15 days starting 3 days postoperatively. The animals were euthanized 3 months postoperatively, and the defect site and the transferred segment were evaluated by radiological and histological examination., Results: The radiological results revealed successful transfers of 14.6 ± 1.2 mm of bone segment on all tibia defects without any complications. The histological evaluation showed new bone formation in both the extension and the docking sites. No rupture or breakage was observed within the plates., Conclusion: We have presented the potential of a new generation ABP for use in segmental bone transfer in an animal model as well as for future clinical applications., Competing Interests: Conflict of interest statement: The authors declare that they have no conflicts of interest., (© TÜBİTAK.)

There is a well-established complex interaction between vitamin D metabolism and bone and gonad functions. In this study, we aimed to investigate the potential effects of vitamin D therapy on testosterone and osteocalcin (OC) levels in aged male rats. Forty-five adult male rats were divided into three groups in this study. Unlike the control group, the two experimental groups received 50 IU/kg/day and 100 IU/kg/day of vitamin D 3 (cholecalciferol), respectively, for a 4-week period using the gavage method. Testicular tissue and blood samples from rats were collected under general anesthesia at the end of the 4-week period. Testicular tissue samples were examined using light and electron microscopy. Additionally, serum testosterone and OC levels were measured in blood samples. The 50 IU/kg dose of cholecalciferol increased testosterone and OC levels, which were lower than normal due to aging, and regulated the organization of the seminiferous tubule epithelium and interstitium more effectively than the 100 IU/kg dose of cholecalciferol. Male fertility functions and bone health, which degrade due to aging, were increased due to the use of exogenous vitamin D, although the higher dose was not associated with more effective results.

The Effect of Pulsed Radiofrequency Application on Nerve Healing After Sciatic Nerve Anastomosis in Rats. In this study, we aimed to evaluate the histomorphological and functional effect of Pulsed Radiofrequency (PRF) application on regeneration after experimental nerve damage in rats. Forty Sprague-Dawley male rats were used in the study. Sciatic nerve incision was applied to all rats and then anastomosis was performed. Twenty rats were separated as the control group, and the remaining 20 rats underwent PRF every day at 42oC, for 120 seconds. The groups were divided into two further subgroups to be sacrificed on the 15th and 30th days. Tissue samples were obtained from all groups at 24 hours and 72 hours after the injury. Sections of sciatic nerve samples were stained with hematoxylin-eosin for light microscopic investigation and prepared for evaluation of ultrastructural changes with transmission electron microscopy. In the evaluation of axon numbers and diameters were seen that the 30th-day RF group had an increase compared to the control group. In the electron microscopic examination, it was observed that myelinated and unmyelinated nerve fiber sheaths had borders that are more regular in the RF group, the nucleus structures of schwann cells were better preserved, mitochondrial damage was less, and the extensions of fibroblast and collagen fibers were smoother than the control group. The findings suggested that PRF application has a positive contribution histologically on nerve healing in the early period after full-layer incision nerve injury anastomosis surgery.

Endometriosis is a common gynecological hurting disorder in which tissue is similar to the tissue that normally lines the inner layer of the uterus. It often causes fertility problems. Unfortunately, effective treatments are limited. Therefore it's important to explore an imperative and easily accessible treatment to alleviate the probable pathologies and preserve fertility in endometriosis. Consequently, we aimed to investigate the effects of metformin, letrozole, and atorvastatin on inflammation and apoptosis in experimentally induced ovarian and peritoneal endometriosis in rat models. In the present study, 35 rats were randomly divided into five groups. Group 1: sham-operated control group. Group 2: untreated endometriosis group. Group 3: given 100 mg/kg/day of oral metformin. Group 4: given 0.1 mg/kg/day of oral letrozole. Group 5: given 2.5 mg/kg/day of oral atorvastatin. At the end of the 28 days, we examined Ki67, Bax and Bcl-2 immunoexpressions in ovarian and peritoneal tissues, and IL-6, IL-8, and TNF-α levels were evaluated from the peritoneal fluid. All medical treatment groups showed a significant decrease in Ki67 expression. A significant increase in Bax expression was also observed in all samples from all medical treatment groups (other than the untreated endometriosis groups). Further, a significant decrease in Bcl-2 expression was found in all medical treatment groups. IL-6, IL-8, and TNF-α levels were significantly lower in all medical treatment groups than in the endometriosis groups. In conclusion; Metformin, letrozole, and atorvastatin showed apoptosis induction and anti-inflammatory effects on both ovarian and peritoneal endometriosis in experimental models., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

This study was aimed at investigating the effects of melatonin, oxytetracycline and N-acetylcysteine on the ovarian follicle reserves and surface epithelium in autologous intraperitoneal ovarian transplantation in rats. Thirty adult female Wistar Albino were selected and randomly divided into six groups (n = 5). Group 1, which was the control group, only had their abdomens opened and closed while Group 2 underwent ovarian transplantation. Group 3, 4, 5 and 6 received 20 μg/kg/IM melatonin, 10 mg/kg/IM oxytetracycline, 150 mg/kg/IP N-Asetil sistein (NAC) and 1% ethanol respectively 15 min before the ovarian transplantation. Vaginal cytology was performed to monitor the estrus phase and the follicle reserve and changes in the surface epithelium were histopathologically evaluated during the preparations. Moreover, cellular apoptosis in tissues was evaluated with immunofluorescence staining of Bcl-2 and Bax. The Bax/Bcl-2 ratio was then calculated as the mean fluorescence intensity (MFI) of Bax and Bcl-2 MFI. Dysplastic change was found only significantly higher in the transplantation group (G2) (p < 0.01). Histopathologically, it was found that the follicle reserve was preserved significantly in the oxytetracycline and melatonin treated group (G3, G4) (p < 0.01). It was also observed that the oxytetracycline treated group (G4) were able to show better preventive effects against dysplastic changes of the surface epithelium. Moreover, the melatonin treated group depicted a low Bax/Bcl-2 ratio compared to the group that only underwent transplantation (G2) (p < 0.01). This study indicated that oxytetracycline and melatonin might be more effective than N-acetylcysteine in protecting against oxidative stress during ovarian transplantation., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

We aimed to create a mechanical optic nerve damage model in rats and to investigate the neuroprotective effects of topical Coenzyme Q10 + Vitamin E TPGS (CoQ10+Vit E) molecule on retinal ganglion cells. In our study, 30 eyes of 20 male Wistar rats were used. Three groups, each consisting of 10 eyes, were formed as control, experimental, and treatment groups. The control group was used to test the formation of optic nerve damage. Topical CoQ10 + Vit E TPGS solution was applied to the rats in the treatment group, one drop twice a day for 3 weeks. On the other hand, physiological drops were applied to the experimental group 2 times a day for 3 weeks. After 3 weeks, the optic nerves of the rats were dissected and examined histopathologically. In electron microscopic examination of the treatment group, it was noted that the myelin sheath in the majority of myelinated nerve fibers and the normal structures of mitochondria, neurotubules, and neurofilaments in the axoplasm were preserved. It was observed that the oligodendrocytes surrounded the myelinated axons. In the experimental group, significant degenerative changes were observed in myelinated nerve fibers in many areas. The number of myelinated axons was significantly increased in the treatment group compared to the experimental group (p = .0028). In the light of the data obtained, the neuroprotective effect of the topically used CoQ10 + Vit E TPGS molecule was found to be histopathologically effective in our experimental study. Abbreviations: NAAION: Nonarteritic anterior ischemic optic neuropathy; CoQ10: Coenzyme q10; CG: Control group; EG: Experimental group; TG: Treatment group.

In this study, the expression of the androgen receptor (AR) and estrogen receptor alpha (ERα) in testicular tissue of male patients with obstructive azoospermia (OA) and non-obstructive azoospermia (NOA) were evaluated by immunohistochemistry. NOA (n = 23) and OA (n = 21) groups were created according to clinical and laboratory archival records. Testicular sperm extraction tissue sections were evaluated according to Johnsen's tubular biopsy scoring (JTBS) method. ERα and AR immunostaining results were evaluated semiquantitatively. Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and estradiol were analyzed. Serum FSH and LH concentrations were greater, and testosterone concentrations were lower than the normal values in the NOA group, whereas the OA group revealed normal hormonal values. Serum estradiol concentrations in groups were in the normal range. JTBSs were significantly lower in the NOA group. Decreased AR expression and increased ERα expression were observed in the NOA group compared to the OA group. This suggests that ERα and AR are expressed in Sertoli cells, Leydig cells, and myoid cells and are required for normal testicular function. Decreased expression of the AR and increased expression of ERα in the testis may negatively affect spermatogenesis. Abbreviations : AR: androgen receptor; ER: estrogen receptor; ERα: estrogen receptor alpha; FSH: follicle-stimulating hormone; JTBS: Johnsen's tubular biopsy scoring; LH: luteinizing hormone; NOA: non-obstructive azoospermia; OA: obstructive azoospermia; TESE: testicular sperm extraction.

Objective: Maternal mood disorders such as postpartum depression (PPD) can negatively affect the lives not only of mothers but also of partners. The purpose of this study investigates emotional behavior and hippocampal apoptosis alterations of the male live with a postpartum depressed female., Methods: Pregnant rats in the stress group were exposed to restraint stress (RS). The male rats who shared the same cages were not exposed to RS. To explain the consequences of depressive-like behavior and anxiety, animals were exposed to the forced swim test (FST), open-field test (OFT), and elevated plus maze (EPM). The apoptotic cell number was detected by terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP biotin nick-end labeling (TUNEL) staining., Results: According to FST, PPD caused more immobility, reduced swimming, and climbing compared to control groups in the stressed female and male (p < 0.05). For the crossing number of squares in the center area, the main effect of the group was significant (p < 0.05). Stressed groups have a higher crossing number of squares in the center area compared to control groups. In the OFT, there was a significant increase in the time spent in the center area in the stress female and male group compared to the control female and male group (p < 0.05). For the EPM, time spent in the close arms was increased in the control male and stress male compared to the stress female group (p < 0.05). Female and male rats with PPD demonstrated apoptosis in neuron and glial cells in the hippocampus., Conclusions: The present study demonstrates that RS results in PPD in females. Furthermore, it implicates RS as a potential risk factor for the development of postpartum mood disorder in males. Most of the studies on paternal PPD have been done by using self-report questionnaires. Studies on physiological and hormonal changes during the postpartum period among fathers would provide information on biological factors of depression., (© 2021 International Society for Developmental Neuroscience.)

Bisphenol A (BPA) is a chemical agent known to have detrimental reproductive and developmental effects. The tissue-specific impacts of BPA exposures and target tissues sensitiveness to BPA are still unclear. The aim of this study was to determine the short- and long-term dose-dependent toxic effects of BPA on rat testes. Forty-eight Wistar albino male rats were divided into four groups each containing 12 rats. To induce toxicity, BPA was administered orally at three different dosages (50, 100, and 200 mg/kg) for 14 and 28 days, respectively. Testis tissues were examined using light and electron microscopy, immunohistochemistry, and biochemical methods. Serum testosterone (T) and luteinizing hormone (LH) levels were measured. Additionally, insulin-like factor 3 (INSL3) as a marker of Leydig cell function was evaluated immunohistochemically. Groups administered high doses of BPA showed severe degenerations such as testicular atrophy, spermatogenic arrest, and interstitial edema in testis. Also, a significant decrease in INSL3 immunoreactivity and serum LH and T levels was found. The results indicated that both increased exposure time and dosage of BPA caused more serious detrimental effects on testes in the rat. Decreased INSL3 and T levels was evidence of Leydig cell function impairment due to BPA.

Peripheral nerve injury (PNI) is a major health problem that results in loss of motor and sensory functions. In treatment of PNI, various methods such as anastomosis, nerve grafts, nonneural tissue grafts, and nerve conduits are applied. In the present study, it was aimed to investigate the effects of Theranekron and Alpha-lipoic acid (ALA) combined treatment on nerve healing in experimental PNI by using histomorphometric, electron microscopic, immunohistochemical and molecular biological methods. Sixty-two Wistar rats were divided into six groups; the normal control group, sham operation group, experimental control group having a crush type injury with no treatment, Theranekron treatment group, ALA treatment group and Theranekron+ALA combined treatment group. Sciatic nerve tissue samples were obtained on days 1, 7 and 14 following injury in all groups. GAP-43 expression was upregulated in all PNI received groups compared to the control group. Krox-20 expression was downregulated in all groups that received PNI compared to the control group. While intensely positive TNF-α and IL-6 expressions were observed up to the 1st to the 14th day for the experimental control group, these expressions were seen as "weakly positive" in the treatment groups from the 1st day to the 14th day. The number of myelinated fibers was higher in the control and sham operation groups. Additionally, the number of myelinated nerve fibers increased in the combined treatment group. In conclusion, these findings suggest that combined therapy of Theranekron and ALA promotes structural recovery and it should be considered as an effective treatment protocol following PNI.

Endometriosis is a gynocological disease characterized by the presence of the endometrial glands and stroma outside the uterine cavity. This disease affects % 6-10 of women with reproductive age and it causes serious problems such as pelvic pain, dysmenorrhea and infertility. Although endometriosis is one of the most investigated disease of gynecology, its pathogenesis is not clear completely. In recent years, many studies revealed the inflammatory nature of endometriosis. Many of the immune cells and their secretory products cytokines and chemokines has been detected in body fluids of women with endometriosis. Cytokines are protein or glycoprotein in structures and hormon-like molecules that act generally in a paracrine fashion to regulate immun responses. They involved in chemotaxis, cell proliferation, cell activation, motility, adhesion and morphogenesis. Tumor necrosis factor alpha (TNF-α) is a proinflammatory cytokine secreted by the macrophages, monocytes, neutrophiles, T cells and natural killer cells. It stimulates increase in the level of the chemokines in body fluids. Monocyte chemotactic protein 2 (MCP-2) is a chemokine act to recruit and activate monocytes into sites of inflammation area. The aim of this study to investigate the ultrastructural properties and whether the expression and localization of TNF-α and MCP-2 in the eutopic endometrium (normal endometrium of women with endometriosis) and endometritic tissues of women with endometriosis. Eutopic endometrial and endometriotic tissue samples were obtained from women with endometriosis between 20-41 y and normal endometrial tissues were collected from 5 women without endometriosis as a control group. Tissues were processed for light and electron microscopy and examined. The epithelial cells of endometriotic tissues were revealed strongly cytoplasmic TNF-α and MCP-2 immunreactivities. Eutopic endometrial tissues were also stained prominently for both TNF-α and MCP-2. Furthermore, a significant increase in stromal macrophages were observed in endometriotic tissues. Moreover, the ultrastructural observations on the normal and endometriotic tissues were exhibited microvilli-rich cells and ciliated cells. These findings suggest that TNF-α and MCP-2 may be involved in normal endometrial biology and in the pathogenesis of endometriosis., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Peripheral nerve injuries (PNI) are an important health problem in the world. In this study, the effects of nerve growth factor (NGF) and betamethasone on nerve regeneration after sciatic nerve crush injury were examined by footprint analysis, electron microscopic, histomorphometric, and biochemical methods. Fifty Wistar rats were divided into five groups as intact control, experimental control, NGF, betamethasone, and NGF+betamethasone combined treatment groups. After the injury, betamethasone was subcutaneously injected into the lesion area of the treatment groups three times during the first day. NGF was subcutaneously injected into the lesion area of treatment groups for 14 days. Footprint analysis was made on 7, 14, 21, 28, and 35 days and after 6 weeks, tissue samples were obtained from all groups. In the experimental control group, there were severe degenerative changes in most of the axons and myelin sheaths of the nerve fibers. Moreover, an increase of MDA levels and a decrease in SOD activities were found in this group. On the other hand, malondialdehyde (MDA) levels decreased, superoxide dismutase (SOD) activities increased and significant motor functional recovery were found in the combined treatment group. The number of axons, axon diameters, and myelin thickness were significantly greater in the combined treatment group when compared with experimental control and other treatment groups. It was thought that nerve regenerative effects of NGF and anti-inflammatory and/or anti-edematous effects of betamethasone could induce functional recovery in the combined treatment group. In conclusion, combined therapy of NGF and betamethasone may be an effective approach for the treatment of PNI.

Spinal cord injury (SCI) is an important health problem, and there is no universal treatment protocol for it today. Following SCI pro-inflammatory mediators such as tumor necrosis factor- alpha (TNF-α) and interleukin-6 (IL-6) increase at the lesion site and play important roles in secondary tissue damage. Methylprednisolone (MP) is a glucocorticoid, and minocycline is a tetracycline-derived antibiotic both with neuroprotective effects on central nervous system trauma. However, there are limited studies on their effects on SCI. In this study, we aimed to evaluate effects of MP+minocycline combined treatment on cellular distribution and localization of TNF-α And IL-6 after SCI. Eighty Wistar rats were divided into three main groups as the intact control group, sham operation group, and experimental control group that received spinal cord compression injury. Following the injury, the experimental control group was subdivided into four groups as control, methylprednisolone treatment, minocycline treatment and, MP+minocycline combined treatment groups. Tissue samples were obtained from all groups at 24 hours and 72 hours after the injury. We found a significant decrease in TNF-α And IL-6 expressions in combined treatment group at 24 hours after injury. Also, there was a significant decrease in MDA and increase in SOD levels in this group. Furthermore, decreased lipid peroxidation and neuronal and glial cell death were also observed in combined treatment group. These results suggest that MP+minocycline combined treatment promotes functional recovery and, it should be considered as an effective treatment protocol following SCI.

Objectives : The aim of the study was to determine the relationships between microRNA-20a and microRNA-125b expression and apoptosis and inflammation in a rat model of spinal cord injury (SCI) using microscopy, immunohistochemistry, and molecular biology. Methods : Sixty-one rats were divided into three groups: a control group that was not subjected to any operation; a sham-operated group; and an experimental group that was subjected to spinal cord compression. The experimental group was further subdivided into two subgroups: the experimental control group, which did not receive any drug treatment; and the methylprednisolone treatment group, which received 30 mg/kg methylprednisolone on day 0 followed by 10 mg/kg/day methylprednisolone from days 1-14. Results : Tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6 levels increased in the experimental control group on days 1 and 3, and decreased in the experimental control group and methylprednisolone treatment group on days 7 and 14. Caspase-3 levels increased in the experimental control group on day 1, and decreased in the experimental control group and methylprednisolone treatment group on days 3, 7, and 14. MicroRNA-20a expression was upregulated in the experimental control group on days 1 and 3, and microRNA-125b expression was downregulated on days 3 and 7. Conclusions : After SCI, upregulated microRNA-20a expression and increased proinflammatory cytokines may lead to an increase in inflammation. MicroRNA-125b may be associated with caspase-3, and microRNA-125b downregulation may inhibit apoptosis. Although the results of this study suggest potential relationships between microRNA-20a and microRNA-125b expression and apoptosis and inflammation in SCI, further studies are needed to confirm microRNA-20a and microRNA-125b as biomarkers in SCI and to develop new strategies for the treatment of SCI.

Proliferation and differentiation of adult Leydig cells are mainly completed in puberty. In many studies, apart from normal postnatal development process, it is widely indicated that, through administrating EDS, Leydig cell population is eliminated and regenerated. It is believed that osteocalcin released from osteoblasts, which is responsible for modulating bone metabolism, induces testosterone production in Leydig cells, independent of the HPG axis. In addition, INSL3 produced by Leydig cells, such as testosterone, plays a critical role in bone metabolism and is known to reflect the development process and functional capacities of Leydig cells. This study is aimed at investigating OC-mediated testosterone regulation and INSL3 synthesis during differentiation of adult Leydig cells that are independent of LH. For this purpose, male rats were divided into 2 groups: prepubertal normal rats and adult EDS-injected rats. Each group was divided into 4 subgroups in which GnRH antagonist or OC was applied. After adult Leydig cells completed their development, testicular tissue samples obtained from the sacrificed rats were examined by light-electron microscopic, immunohistochemical, and biochemical methods. Slight upregulation in 3 β HSD, INSL3, and GPRC6A expressions along with the increase in serum testosterone levels was observed in groups treated with osteocalcin against GnRH antagonist. In addition, biochemical and microscopic findings in osteocalcin treated groups were similar to those in control groups. While there was no significant difference in the number of Leydig cells reported, the presence of a significant upregulation in INSL3 and GPRC6A expressions and the increase in serum testosterone and ucOC levels were observed. After evaluation of findings altogether, it is put forward that, for the first time in this study, although osteocalcin treatment made no significant difference in the number of Leydig cells, it increased the level of testosterone through improving the function of existing adult Leydig cells during normal postnatal development process and post-EDS regeneration. This positive correlation between osteocalcin-testosterone and osteocalcin-INSL3 is concluded to be independent of LH at in vivo conditions., Competing Interests: The authors declare no conflicts of interest.

Purpose: To investigate changes in conjunctival tissue of conjunctivochalasis (CCh) patients and to determine the relationship between pathological findings and localization of loose conjunctiva., Methods: Our study included nineteen eyes of 19 patients who were referred to Cukurova University Ophthalmology Department based on ocular surface symptoms and CCh detected in ocular examination. Amniotic membrane was applied after conjunctival excision as surgical treatment. The control group was formed with five eyes of five patients who are similar in terms of age and gender distribution with our study group. Tissue samples obtained from the study and control groups were investigated with light and electron microscopy., Results: Results of pathological examination of conjunctival tissues revealed increased inflammation in 13 patients (68%), lymphatic ectasia in 12 patients (63%), and loss of goblet cells in 17 patients (89%). Destruction of elastic fibers was detected in all cases by staining with elastic van Gieson. After semiquantitative assessment, varying degrees of light microscopic findings were noted considering the localization of CCh. No statistically significant relationship was observed between light microscopic findings and CCh location (p > 0.05 for all). Electron microscopic investigation revealed increase in intercellular spaces, increased cytoplasmic electron density, and the presence of slight vacuolization in cell cytoplasm, and heterochromatin clumping in nuclei of cells in conjunctival samples., Conclusions: Mechanical and inflammatory factors induce development of CCh, and signs associated with these factors can be detected with light and electron microscopy of conjunctival tissue. No relationship was observed between CCh localization and pathological changes in tissues examined in our study, and large-scale case series are required to evaluate the possible effect of CCh localization on pathological findings.

Introduction: The cytokine profile and the ultrastructural changes of refluxing ureterovesical junctions(UVJs) of children treated with failed dextranomer/hyaluronic-acid (Dx/HA) injections were investigated using immunohis-tochemical methods and transmission electron microscopy(TEM)., Patients and Methods: Eighteen children who had undergone injection for reflux were included the study. The smooth muscle arrangement of the ureteral wall, transforming growth factor-? (TGF-?1),vascular-endotheli-al-growth factor (VEGF) and CD34 were evaluated immunohistochemically, and the results were compared with 10 age-matched autopsy specimens as controls. The ultrastructural evaluation and morphological description was made semi-quantitatively and compared with published data., Result: Four of the patients (22%) were male, and 14 (78%) were female. The mean age of the patients was 105.4 ± 44.5(48-184) months. There was no correlation between the vesicoureteral reflux (VUR) grade and age (P = 0.85). The mean VEGF and CD34 scores were 16.2 ± 9.6 (0-90) cells per HPF and 10.2 ± 3.5 (4-16) vessels per HPF in ureters with reflux; these values were 60.6±16.4 (32-84) cells per HPF and 17.8 ± 4.1 (12-24) vessels per HPF in the control group. The amount of VEGF and CD34 were significantly decreased in patients compared with the control group (P < 0.001, P < 0.001).The TGF-?1 levels were significantly higher in patients with VUR compared with the control group (34.2 ± 19.9 vs 5.0±1.9; P=0.001).The amount of VEGF, CD34, and TGF-?1 were not correlated with the grade of reflux (P = 0.26, P = 0.94, and P = 0.42, respectively). Ultrastructural changes in the muscle cells were observed in all the VUR specimens (Grade II-IV)., Conclusion: Refluxing ureters exhibited immune-histopathological abnormalities and ultrastructural changes of the muscle cells in all VUR specimens in the ureterovesical junctions of children treated with failed Dx/HA injec-tions for reflux.

Hydrosalpinx is a disease commonly observed in women and characterized by the obstruction which is in the shape of a fluid-filled sac at the distal part of tuba uterina closed to the ovary. In this study, we aimed to obtain endometrial tissue samples from the hydrosalpinx patients, before and after the surgical treatment and compare these endometrial tissue samples by using light and electron microscope. Endometrial tissue samples were obtained from the 24 women with bilateral hydrosalpinx range 19-46 years before and after the surgical treatment, and normal endometrial tissues were collected from five women without hydrosalpinx and evaluated as a control group. In endometrial samples of hydrosalpinx patients; it was observed that large and unregulated interstitial spaces representing the organellar destruction, membranous whorl structures associated with organelle destruction, thinning in the surface epithelium, decreasing in numbers of microvillus and pinopodes in microvilli cells, increasing in heterochromatin and picnotic changes in the nucleus, expansion, and vacuolization in the endoplasmic reticulum cisternae in the apical cytoplasm and intraepithelial macrophages and lymphocytes were rised in number. Although mild structural changes were observed in endometrial tissues obtained after surgical treatment of hydrosalpinx, surface epithelium, glandular and stromal cell structures were more similar to control endometrial specimens. In conclusion; serious structural changes have occurred in endometrial tissues of hydrosalpinx patients. These structural abnormalities have removed after surgical treatment so it is considered that surgical treatment is effective in patients with hydrosalpinx.

Background: A single dose injection or continuous infusion of local anesthetics into the joint space is considered to be a well-defined analgesia technique. The aim of this study was to investigate the chondrotoxic and apoptotic effects of single-dose intra-articular injection of levobupivacaine and bupivacaine on rabbit knee joint tissues., Materials and Methods: The animals were allocated into two groups each containing 20 rabbits. 0.5% levobupivacaine (Group L) and 0.5% bupivacaine (Group B) were applied intra-articularly to the left posterior joints of rabbits. At the same time, normal saline was applied to the right posterior leg knee joints of rabbits in both groups and used as a control (Group S). At the end of the 7th and 28th days after the intraarticular injections, ten randomly chosen rabbits in each group were killed by applying intraperitoneal thiopental. Sections of cartilage tissue samples were stained for light microscopic examinations and the TUNEL method was used to investigate apoptotic cells., Results: As a result of immunofluorescence microscopic examination, the number of apoptotic cells in Group B at day 7 and day 28 were both significantly higher than Group L and S (p<0.05). Also, the number of apoptotic cells in Group L at day 7 and day 28 were both significantly higher than Group S (p<0.05)., Conclusions: We found that bupivacaine is more chondrotoxic than other anesthetic agent and increases the number of apoptotic cells. These results indicated that bupivacaine caused high chondrotoxic damage and it led to more apoptotic activation than levobupivacaine., (Copyright © 2018 Sociedade Brasileira de Anestesiologia. Publicado por Elsevier Editora Ltda. All rights reserved.)

Exposure to cigarette smoke (CS) causes vessel damage and mechanism of this damage has not yet been clearly identified. Therefore, in this study we aimed to investigate whether vessel damage due to the CS exposure will be prevented by the alpha-linolenic acid (ALA) or not which has anti-inflammatory effect in mice. For this reason, mice were grouped as controls (with and without CS) and ALA (with and without CS). The CS application continued 5 days a week for two months. At the end of two months, the mice were killed by cervical dislocation and their blood and thoracic aortas were isolated. ALA Treatment increased acetylcholine relaxations. CS decreased acetylcholine relaxation. CS with ALA treatment increased acetylcholine relaxations versus just CS treatment. CS caused rising in cyclooxigenase-2 and phospholipase A2 levels. This rise is inhibited with ALA treatment. CS decreased eNOS levels. But this result was not statistically significant. Furthermore, according to electron microscopic study CS damaged both smooth muscle and endothelium. While ALA treatment prevented smooth muscle damage it didn't prevent endothelial damage. Using cigarette and CS exposure is a risk factor for cardiovascular disease. Our study showed that this disease., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Aims: The aim of this study was to investigate the effects of pterostilbene (PTS) (trans-3,5-dimethoxy-4'-hydroxystilbene) and resveratrol (RSV) (trans-3,5,4' trihydroxystilbene) applied at different doses for the treatment of streptozotocin- (STZ-) induced diabetic rats., Materials and Methods: At the end of the 5-week experimental period, the right gastrocnemius muscles of the rats were examined biomechanically, while the left ones were examined histologically. In addition, blood glucose, serum insulin, and malondialdehyde (MDA) levels were analyzed in blood samples taken from the rats., Results: The skeletal muscle isometric contraction forces, which showed a decrease with diabetes, were observed to increase with antioxidant applications. Blood glucose, serum insulin, and MDA levels in diabetic rats approached normal levels after applying PTS. When the electron microscopic images of the rat skeletal muscle were examined, those in the combination treatment group were observed to show a better enhancement in the skeletal muscle morphological structure compared to the other diabetic and treatment groups., Conclusion: According to the findings, we suggest that these antioxidant treatments might have good therapeutic nutraceutical potential for some muscle diseases that coexist with diabetes. These treatments should be comprehensively investigated in the future.

Recent studies suggest that nerve growth factor (NGF) protects olfactory cells and axons from injury in vitro. Eighteen Wistar-Albino rats randomly divided into three groups: control group, diabetic group without NGF, and diabetic with NGF. Intranasal NGF (6 µg/day) was administered over a 5-day period. At the end of 30 days, the olfactory epithelium (OE) of NGF-applied diabetic rats regenerated, the epithelium thickness was significantly higher, and caspase-3 expression was not significantly different from the control. The current results demonstrate that intranasally administered NGF significantly reversed OE morphological changes in diabetes by decreasing diabetes-related cell death and inflammation.

Purpose: Subchondral perfusion of osteochondral grafts has been shown to be important in preventing long-term cartilage degeneration. In carpal reconstruction, subchondral perfusion from the graft bed is limited. This study's purpose was to compare the histological characteristics of cartilage in osteochondral grafts supported by synovial imbibition alone to cartilage of vascularized osteochondral flaps that have both synovial and vascular pedicle perfusion., Methods: Two adjacent osteochondral segments were harvested on the medial femoral trochlea in domestic 6- to 8-month-old pigs. Each segment measured approximately 12 mm × 15 mm × 17 mm. One segment was maintained on the descending geniculate artery vascular pedicle. The adjacent segment was separated from the pedicle to serve as a nonvascularized graft. A thin layer of methylmethacrylate cement was used to line the harvest site defect to prevent vascular ingrowth to the subsequently replaced specimens. The pigs were maintained on a high-calorie feed and returned to ambulation and full weight-bearing on the surgical legs. The animals were sacrificed after 6 months and the specimens were reharvested, sectioned, and examined. The cartilage was graded by 2 pathologists blinded to the origin of specimens as vascularized flaps or nonvascularized grafts., Results: All specimens were assigned scores utilizing the International Cartilage Repair Society grading system. Scoring for chondrocyte viability, cartilage surface morphology, and cell and matrix appearance was significantly higher in the vascularized osteochondral group than in the graft group., Conclusions: When deprived of subchondral perfusion from underlying bone, osteochondral vascularized flaps in an intrasynovial environment demonstrate superior cartilage quality and survival compared with nonvascularized grafts., Clinical Relevance: In locations in which perfusion from surrounding bone may be limited (ie, proximal scaphoid or proximal lunate reconstruction), articular reconstruction using vascularized osteochondral flaps will yield superior cartilage organization and architecture than nonvascularized osteochondral grafts. The clinical and functional relevance of this finding requires further study., (Copyright © 2018 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.)

Aim: To examine morphological, radiological and biochemical effects of arginine vasopressin (AV) and V1 receptor antagonist on cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) in rabbits., Material and Methods: Forty male New Zealand white rabbits were randomly divided into four groups comprising 10 rabbits each. The groups were; 1) Control group, 2) SAH group, 3) SAH+AV group, 4) SAH+V1 antagonist group. Diameters of the basilar artery in all groups were measured on angiograms. All animals were sacrificed two days following basilar angiography and tissue samples of basilar artery were obtained under microscope immediate after craniectomy for ultrastructural and biochemical examinations., Results: The artery diameters were found to be 50% and 50% at the 30th minute in the groups 2 and 3 respectively. In group 3, CVS was 13% more in comparison with the 2nd group. In group 4, vascular constriction was 34.5% at the 30th minute and about 30.9% at the 300th minute. Despite the increase in regional blood flow, AV did not provide morphological change. Histological appearance was related to vascular stenosis due to CVS. Histological outcome was the best in group 4 because of less CVS., Conclusion: Arginine vasopressin plays an important role in CVS. We detected morphological and radiological recovery in basilar artery, besides moderate improvement due to AV receptor antagonist in CVS.

Background/aim: The aim of this study was to determine the antioxidative effect of curcumin on nicotine-induced mice testis., Materials and Methods: Sixty Swiss albino male mice were divided into five groups, each containing 12 mice. The first group was used as a control. To induce toxicity in the second and third group, nicotine (0.4 mg/kg/day) was injected intraperitoneally into mice for 14 and 28 days, respectively. The mice in the fourth and fifth group were injected with nicotine (0.4 mg/kg/day) and orally treated with curcumin (200 mg/kg) for 14 and 28 days, respectively. Testosterone levels were measured from blood samples and testis tissues were examined under light and electron microscopes., Results: Light and electron microscopic examinations of the nicotine-induced groups showed evident degenerations in spermatogenic cells, Sertoli cells, and Leydig cells. The groups treated with curcumin had less testicular alterations. The mice that were sacrificed after 28 days in the groups treated with curcumin showed minor degenerations. Furthermore, the median levels of testosterone significantly decreased in the nicotine-induced groups in comparison with those in the control group., Conclusion: The results indicated that curcumin might be a potential therapeutic agent for testicular injury caused by nicotine addiction.

Oxidative stress is one of the main causes of diabetic nephropathy, which is a complication of diabetes mellitus (DM). The aim of this study was to investigate the possible role of ethyl pyruvate (EP) in streptozotocin-induced diabetic rats' kidney. Four groups (n = 8) of male Wistar albino rats were used as follows: control group rats received only sodium citrate buffer solution intraperitoneally (ip). The EP group was given 50 mg/kg EP ip. In the DM group, diabetes was induced by streptozotocin. The DM + EP group received 50 mg/kg EP ip. All animals received daily treatment for 14 days, and at the end of the study the kidneys were removed: the left kidney of the rats was used for malondialdehyde (MDA) analysis and the right kidney for histological examination. There was normal appearance of the kidney tissues in the control and the EP-administered groups. In the DM group, there was evident basement membrane thickening and enlargement of mesangial matrix; swelling in some tubular epithelial cells was also noticeable. In the DM+EP administered group, nearly the same appearance as the control group and relative thickening in the glomerular basal membrane were observed. The antioxidant effect of ethyl pyruvate improved the renal structures in the DM + EP group.

Objective: Necrotizing enterocolitis has been investigated and debated extensively in recent years; however, there is still no effective treatment. The aim of this study was thus to examine the effects of β-estradiol on intestinal injury in rats., Methods: Twenty-four newborn female rat pups were divided into three groups. In group 1 (sham), hypoxia-re-oxygenation was not performed. In group 2 (saline), the rats were injected with saline after hypoxia-re-oxygenation, and the process was repeated for 5 d. In group 3 (β-estradiol treatment), the rats were subjected to hypoxia-re-oxygenation and then given β-estradiol intraperitoneally once a day for 5 d. After these procedures, the terminal ileum was removed for analysis., Results: Statistically significant differences in histological grades were found between groups 1 and 2 (p = 0.000), groups 1 and 3 (p = 0.028), and groups 2 and 3 (p = 0.021). There were also differences in TNF-α and IL-6 levels between groups 2 and 3 (p = 0.000 and p = 0.038, respectively) and between groups 1 and 2 (p = 0.000 and p = 0.000); there was no difference between groups 1 and 3 (p = 0.574 and p = 0.195, respectively). Electron microscopy examination revealed a decrease in lipid droplets at the apical cytoplasm of the columnar cells in group 2; in group 3, the absorption of the lipids as lipid droplets was similar to that of group 1., Conclusion: In this study, β-estradiol was found to decrease the intensity of intestinal injury significantly by inhibiting TNF-α and IL-6.

The aim of this study was to investigate the myotoxic effects of bupivacaine, ropivacaine, and levobupivacaine which were applied intramuscularly to rat skeletal muscle. Forty Wistar-Albino rats were divided into four groups. In the study, .5% bupivacaine (Group B), .5% ropivacaine (Group R), .5% levobupivacaine (Group L), or .9% normal saline (Group SF) was applied intramuscularly to the right gastrocnemius muscle of rats. The rats in each group were sacrificed on the second day after injection. Sections of muscle samples were stained with hematoxylin-eosin for light microscopic investigation and prepared for the evaluation of ultrastructural changes in the subcellular level with transmission electron microscopy. All three local anesthetic agents caused qualitatively similar skeletal muscle damage. The most observed muscle damage was in Group B, muscle damage of Group R was less than that of Group B, and the least damage was seen in Group L quantitatively. Electron microscopic examination of each group that caused cellular damage was qualitatively similar. The most subcellular damage was observed in the group receiving bupivacaine, less was seen in the ropivacaine group, and the least was observed in the levobupivacaine group. The results indicated that bupivacaine caused more myotoxic damage than the other two agents in the skeletal muscle of rats and that levobupivacaine caused less myotoxic damage than both bupivacaine and ropivacaine at the cell and tissue levels.

Purpose: We used immunohistochemical methods and transmission electron microscopy to investigate the cytokine profiles and ultrastructural changes in the ureterovesical junction of children with primary vesicoureteral reflux., Materials and Methods: A total of 39 distal intravesical ureters were obtained from 23 children who underwent ureteroneocystostomy for primary vesicoureteral reflux. Ureteral wall smooth muscle organization and transforming growth factor-β1, vascular endothelial growth factor and CD34 were evaluated immunohistochemically and compared to controls, which consisted of 10 age matched autopsy specimens. Ultrastructural evaluations and morphological descriptions were performed semiquantitatively and compared to the published data., Results: Of the patients 6 (26%) were male and 17 (74%) were female, and mean ± SD age was 73.2 ± 34.3 months (range 12 to 168). There was no correlation between reflux grade and age (p = 0.39). Smooth muscle disorganization score differed significantly between patients with intravesical ureters and controls (p = 0.01). Transforming growth factor-β1 levels were significantly higher (p = 0.001) and vascular endothelial growth factor levels and microvessel densities were significantly lower in the patients with reflux compared to controls (both p <0.001). Vascular endothelial growth factor, CD34 and transforming growth factor-β1 levels did not correlate with reflux grades (p = 0.84, p = 0.76 and p = 0.10, respectively). Urothelium, lamina propria and tunica adventitia appeared normal in the specimens for all grades of vesicoureteral reflux using transmission electron microscopy. Damage was observed in the muscular layers of the ureterovesical junction, especially in patients with grade IV or V reflux., Conclusions: Primary refluxing ureters exhibit immunohistopathological abnormalities compared to normal ureters irrespective of reflux grade, and ultrastructural changes are especially severe in cases of high grade reflux. These abnormalities can hinder the normal ureteral valve mechanism, and may lead to reflux due to smooth muscle dysfunction and microvascular alterations., (Copyright © 2014 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

Objective: The objective of this study was to compare the histological regeneration characteristics of nerve fibers at the anastomosis lines performed by classic suture technique or a tissue adhesive (N-butyl-2-cyanoacrylate)., Methods: The control group consisted of 7 rabbits. The 21 rabbits were randomly divided into 3 groups based on the harvesting week. In the study group following preparation of facial nerve bilaterally, a 0.5-cm segment of facial dorsal buccal nerve was resected, and the defect was repaired with a nerve graft, which was harvested from sural nerve of the same side by 8-0 nylon suture technique and by application of N-butyl-2-cyanoacrylate on the other side., Results: Electron microscopic examination at consecutive second, fourth, and sixth days (corresponding to 4th, 8th, and 12th week in human subjects) revealed increased nerve degeneration findings in N-butyl-2-cyanoacrylate group when compared with microsuture repair technique., Conclusions: We conclude that N-butyl-2-cyanoacrylate is not an appropriate material for nerve anastomosis.

Phenolic compounds make up the major secondary metabolites with high pharmaceutical potential. Microalgae were reported to contain low amounts of phenolic compounds. The present study aimed to investigate the hepatoprotective potential of biomass of Spirulina platensis enriched in phenolic compounds. The protective effects of the biomass of S. platensis with low amounts of phenolics (SP1) and with high amounts of phenolics (SP2) against CCl4-induced acute hepatotoxicity were evaluated in rats. The increased levels of ALT, AST and MDA along with decreased activities of SOD and CAT were significantly (p<0.01) ameliorated by SP2. Histological examinations revealed that SP2 was more potent than SP1 in protecting the liver from toxic injury of CCl4 and preserving the hepatocyte ultrastructure. The lesions including necrosis, lymphocyte infiltration, ballooning degeneration and hepatocyte injury as irregular lamellar organisation, dilations in endoplasmic reticulums and the presence of great number of cytoplasmic vacuolization were healed by SP2., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The aim of this study is to investigate the possible protective effect of N-Acetylcysteine (NAC) against the likely methotrexate (MTX) toxicity on the kidney using ultrastructural together with biochemical data. Moreover, the immunohistochemical detection of Ki67 nuclear antigen is to be evaluated. Fifteen male Wistar albino rats, weighing 240-290 g, were divided into three equal groups: Rats receiving MTX alone, rats receiving MTX plus NAC treatment, and rats comprising the control group. MTX (18 mg/kg/day, body weight) in dissolved physiologic saline was administered intraperitoneally to rats during 3 days. For the MTX plus NAC group, N-Acetylcysteine (300 mg/kg/day, body weight) was administered together with MTX. At the end of the third day, all the rats were killed with cervical dislocation to obtain blood and tissue samples. Application of MTX principally induced prominent large vacuolization in the proximal convoluted tubule cells, and focal thickening in the glomerular basal lamina of some glomeruli. A decrease in tissue SOD (superoxide dismutase) and GSH-Px (glutathione peroxidase), and an increase in serum urea nitrogen and creatinine and in tissue MDA (malondialdehyde) levels were also seen in the MTX group. These changes were significantly reversed in the MTX-plus-NAC-treated group. Most of the vacuoles in the proximal convoluted tubule cells disappeared. Furthermore, an increase in antioxidant enzyme activities, a decrease in serum urea nitrogen and creatinine, and tissue MDA levels were all significant. Additionally, an increase in the number of Ki67 positive-stained cells in proximal tubules was also noted. In conclusion, NAC may be a promising substance against MTX-induced renal damage. It might be useful to use NAC supplementally to minimize MTX-induced nephrotoxicity.

Purpose: To investigate the ultrastructural changes of the rabbit retina induced by intravitreal methotrexate injection., Materials and Methods: Ten New Zealand white rabbits were enucleated bilaterally at different time periods after intravitreal methotrexate injection. One rabbit was used as control group and one rabbit was used as intact group. Histopathological examinations were performed under light and electron microscopy. Early (within first three days after injection) and long-term (one month after serial injections) effects of intravitreal methotrexate on the retina were investigated., Results: Retinal edema, vacuolization, and disintegration of mitochondria of the retinal cells were observed as early changes. The main long-term effects after serial injections were edema in the photoreceptor, inner nuclear, and ganglionic cell layers. Cellular disorganisation was seen on light microscopy. Electron microscopic examination revealed mitochondrial degeneration and vacuole formation in retinal cells, nuclear degeneration in outer nuclear layer, and membranous whorl formation in photoreceptor and nerve fiber layers., Conclusions: High dose intravitreal methotrexate injection may cause significant ultrastructural changes in the rabbit retina in varying severity. This finding may highlight the potential side effects of methotrexate on human retina in higher doses.

The N-Methyl-D-Aspartate (NMDA) receptor is expressed abundantly in the brain and plays an important role in neuronal development, learning and memory, neurodegenerative diseases, and neurogenesis. In this study, we evaluated the effects of NMDA receptor blockade during the early neurodevelopmental period on exploratory locomotion, anxiety-like behaviors and cognitive functions of adolescent Wistar rats. NMDA receptor hypofunction was induced 7-10 days after birth using MK-801 in rats (0.25 mg/kg twice a day for 4 days via intraperitoneal injection). The open-field (OF), elevated plus maze (EPM) and passive avoidance (PA) tests were used to evaluate exploratory locomotion, anxiety-like behaviors and cognitive functions. In the OF test, MK-801 caused an increase in locomotion behavior (p < 0.01) and in the frequency of rearing (p < 0.05). In the EPM test, MK-801 treatment increased the time spent in the open arms, the number of open arm entries and the amount of head dipping (p < 0.01). MK-801 treatment caused no statistical difference compared to the control group in the PA test (p > 0.05). Chronic NMDA receptor blockade during the critical period of maturation for the glutamatergic brain system (postnatal days 7-10) produces locomotor hyperactivity and decreased anxiety levels, but has no significant main effect on cognitive function during adolescence.

Objective: We examine the ultrastructural configurations of Cajal cells by electron microscopy, as well as the quantitative changes occurring in Cajal cells by light microscopy., Methods: In total, 35 patients with ureteropelvic junction (UPJ) obstruction and 7 patients without obstruction were compared immunohistochemically with c-kit (CD117) to quantify the number of cells. On electron microscopic examination, 7 patients with UPJ obstruction and 3 patients without obstruction were compared to evaluate the changes which occurred in the ultrastructural configuration of the Cajal cells., Results: On light microscopic examination, it was determined that the Cajal cells, which demonstrate c-kit (CD117) immunoreactive character, were located near the circular muscle layer and parallel to the muscle cells. The number of Cajal cells in the control group was significantly increased compared to the number of cells in patients with UPJ obstruction (p < 0.001). On electron microscopic examination, the number of interstitial cells was also higher in the control group. A decrease in the number of the caveolae in these cells was seen in the group with UPJ obstruction compared to the control group., Conclusion: In UPJ obstruction, a decrease in the number of Cajal cells, as well as the changes in the morphologic structure of the Cajal cells, indicates that these cells have a role in the pacemaker system and are associated with ureteral peristalsis.

Background: Hypoxic-ischemic brain injury (HIBI) is a common cause of neonatal mortality and morbidity. The use of levetiracetam (LEV), as a potential neuroprotective in brain ischemia, receives an increasingly high attention, and it could have a crucial role in the regulation of epileptogenesis and neuroprotection. Potential effects of LEV on neuronal apoptosis in HIBI have not previously been reported in literature., Objectives: The aim of this study is to evaluate the possible effects of LEV on neuronal apoptosis in neonatal rat model of HIBI., Methods: Seven-day-old Wistar rat pups were subjected to right common carotid artery ligation and hypoxia (92% nitrogen and 8% oxygen) for 2h. The pups were treated with LEV or saline after hypoxia. In sham group rats, neither ligation, nor hypoxia was performed. Neuronal apoptosis was evaluated by the terminal deoxynucleotidyl-transferase- mediated dUTP nick-end labeling (TUNEL) methods., Results: The counts of apoptotic cells in both hippocampus and cerebral cortex were significantly higher in the saline treatment group than in the sham group. The counts of apoptotic cells in both hippocampus and cerebral cortex were similar to those in the sham group and in the LEV treatment group. The number of apoptotic cells decreased significantly in the LEV-treated group compared with the saline group., Conclusions: These results show that LEV administration after hypoxia reduces neuronal apoptosis. Thus, we propose that LEV, as an effective antiepileptic and antiapoptotic drug, may be a viable choice for the control of seizure activity in neonates with HIBI., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Introduction: The management of acute respiratory distress syndrome (ARDS) was investigated with the use of heliox in an experimental model., Objectives: To investigate whether heliox can be considered a new therapeutic approach in ARDS., Methods: ARDS was designed in Wistar albino male rats, 250-300 g in weight, by intratracheal instillation of physiological saline solution. Anesthezied and tracheotomized rats with ARDS were pressure-controlled ventilated. At the end of 210 min, helium gas was tried. All rats were assigned to two groups: Group 1 (n = 10) was the control group, and was given no treatment; group 2 (n = 7) was given heliox (He: O(2) = 50:50). The heliox group received heliox for 1 h continously. Rats were continued to be kept on a ventilator through the experiment. Two hours after the last inhalation, both lungs of the rats were excised for both histopathological examination and immunohistochemical evaluation., Statistical Analysis Used: Histopathological grading were expressed as median interquartile range. Mann-Whitney U-test was used to assess the relationships between the variables., Results: The infiltation of neutrophils were decreased in rats treated with heliox. Edema in the interstitial and intraalveolar areas was less than that of the control rats. Also, the diminishing of perivascular and/or intraalveolar hemorrhage was apperant. Hyaline membrane (HM) formation decreased in the heliox group compared with the control group. Decreased inducible nitric oxide synthase expression was shown via immunohistochemical examination in the heliox group., Conclusion: The present study histopathologically indicated the effectiveness of heliox in the decreasing of neutrophil infiltation, interstitial/intraalveolar edema, perivascular and/or intraalveolar hemorrhage and HM formation in ARDS. Besides the known effect of heliox in obstructive lung disease, inhaled heliox therapy could be associated with the improvement of inflamation in ARDS.

Objective: To investigate whether the low-intensity electromagnetic waves transmitted by cell phones cause histopathological or ultrastructural changes in the testes of rats., Materials and Methods: Wistar-Kyoto male rats were placed into either a control group or a group that was exposed to an electromagnetic field (EMF). Two cell phones with Specific Absorbation Rate values of 1.58 were placed and left off in cages that housed 15 rats included in the control group, and four cell phones were placed and left on in cages that housed 30 rats included in the experimental group. After 3 months, weights, seminiferous tubule diameters, and spermatogenic cell conditions of all testes of the rats were evaluated. One half of each testis was examined also under an electron microscope., Results: No significant differences were observed between the testis weights, seminiferous tubule diameters, and histopathological evaluations between rats that had and had not been exposed to EMF. Electron microscope analysis revealed that the membrana propria thickness and the collagen fiber contents were increased and the capillary veins extended in the experimental group. Common vacuolization in the cytoplasm of the Sertoli cells, growth of electron-dense structures, and existence of large lipid droplets were noted as the remarkable findings of this study., Conclusion: Although the cells that had been exposed to long-term, low-dose EMF did not present any findings that were contrary to the control conditions, the changes observed during ultrastructural examination gave the impression that significant changes may occur if the study period were to be extended. Longer studies are needed to better understand the effects of EMFs on testis tissue., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Purpose: The purpose of the present study was to evaluate the effects of systemically administered zoledronic acid (ZA) on mandibular fracture healing in a rabbit model using radiodensitometric, biomechanical, histologic, and histomorphometric methods., Materials and Methods: A total of 36 skeletally mature male New Zealand white rabbits were used. The rabbits were randomly divided into 2 groups. A mandibular corpus fracture was created experimentally in all 36 rabbits. The experimental group was administered an intravenous, single dose of 0.1 mg/kg ZA, and the control group was administered only saline infusion during the procedure. All rabbits were sacrificed on the 21st postoperative day. Digital radiodensitometric analysis, a 3-point bending test, and histologic and histomorphometric examinations were performed on the harvested hemimandibles. The data were analyzed statistically., Results: Biomechanical testing data showed that ZA treatment resulted in a significant increase in the healed bone strength. This result was supported by the radiologic, histologic, and histomorphometric findings., Conclusions: The results of the present study have revealed that systemic administration of ZA accelerates and improves the bone healing of mandibular fractures., (Copyright © 2011 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

Object: Fresh autogenous bone graft is the most preferred osteoplastic material, whether the purpose is cosmetic, psychological, or for the protection of the brain. These grafts are not rejected and do not react immunologically. The aim of this study was to evaluate the efficacy of autogenous fat rolled with bone dust derived from the bur hole in closing small cranial defects. Additionally, the authors examined the morphological and biochemical effects of Na selenite and amiloride on calvarial calcification., Methods: The study group consisted of 20 domestic pigs. These animals were randomly divided into 4 groups. A bur hole with a diameter of 10 mm was made at the right parietal region in all animals, and then the periosteum around the bur hole was cauterized following exposure of the dura mater. The dura was coagulated with bipolar cautery. Group 1 (controls): only a bur hole was opened, and it was then closed with a mixture of the bone dust that had been created during the opening of the bur hole and fat tissue that was taken from the animal's neck. Group 2 (amiloride): 1 nmol/g body weight of amiloride was applied subcutaneously within 15 minutes after closure of the bur hole with bone dust and fat, and then amiloride was applied once a day for 4 weeks. Group 3 (Na selenite): 30 nmol/g body weight of Na selenite was applied subcutaneously within 30 minutes after closure of the bur hole with bone dust and fat, and then Na selenite was applied once a day for 4 weeks. Group 4 (amiloride and Na selenite): 1 nmol/g body weight of amiloride was applied subcutaneously at 15 minutes, and 30 nmol/g body weight of Na selenite was applied subcutaneously at 30 minutes after closure of the bur hole with bone dust and fat, and these 2 injections were repeated once a day for 4 weeks. At the end of 4 weeks, the animals were anesthetized to evaluate the closure of the bur hole. Tissue samples were obtained for ultrastructural and biochemical examination., Results: The defect was covered with diffuse connective tissue in the control group. Although multiple capillary vessels were present, the authors did not observe osteogenic differentiation. Histological examination of the second group revealed osteogenic changes. Although new matrix was formed, calcification was not detected. The authors observed fibroblast, collagen fibers, and dense connective tissue filled with capillary in the third group of pigs, which had undergone Na selenite application. Calcification was not detected in this group. Both connective and osteogenic tissue were observed in specimens obtained in the fourth group, which had undergone amiloride and Na selenite application., Conclusions: The authors experimentally evaluated the supplementary osteogenic effects of Na selenite and amiloride by using them separately and together. The findings seem promising as a lead-in to new studies in restoring cranial defects.

Objectives: Our aim was to investigate whether neutralization of rat interleukin 6 (IL-6) bioactivity attenuates inducible nitric oxide synthase (iNOS) up-regulation and ameliorates cerebral ischemic damage in a model of focal central nervous system (CNS) ischemia., Methods: Seventy rats were randomly allocated to groups: Group I (n=10) consisted of normal controls; Group II (n=20) underwent surgical exposure of the middle cerebral artery but no cauterization; the remaining 40 rats were subjected to middle cerebral artery occlusion. Immediately after occlusion, each of these 40 rats was randomly assigned to either the occlusion-only group (Group III, n=20) or the occlusion plus IL-6 antibody treatment group (Group IV, n=20). Half of the rats from each of Groups II, III and IV were eternized at 24 hours and the other half at 72 hours. The samples were used for iNOS immunohistochemistry and structural analysis., Results: A single dose of the antibody had no effect on structural changes and iNOS at 24 hours after occlusion. However, administering three doses of the antibody resulted in markedly decreased quantitative and qualitative levels of iNOS-positive stained cells and milder subcellular damage compared with the findings in the occlusion-only group at 72 hours after occlusion., Discussion: Our findings prove that IL-6 bioactivity is one of the pathological events that trigger the induction of iNOS in the process of CNS ischemic injury. It appears that there may be therapeutic value in neutralization of IL-6 bioactivity to attenuate iNOS up-regulation and ameliorate cerebral ischemic damage in long-term recovery.

Purpose: The aim of this study was to compare the vascularized osteoperiosteal femur flaps (VFFs) and the nonvascularized femur grafts (NVFGs) for reconstruction of the mandibular defects of pigs., Materials and Methods: Eight adult domestic pigs were used. The defects created in the mandibular angle were reconstructed with VFFs in 4 pigs (group 1) and NVFGs in the other 4 pigs (group 2). All the pigs were killed after 3 months of healing. Undecalcified and decalcified sections were prepared for histomorphometric analysis and histologic examination. Radiodensitometric absorptiometry was used to assess the differences in bone mineral density between the 2 groups., Results: The bone volume to the total measured volume, trabecular thickness, and trabecular number were significantly greater in the VFF group than in the NVFG group (P < .05). However, the trabecular separation was significantly lower in the VFF group than the NVFG group (P = .029). Although the VFF group had a greater bone mineral density value than the NVFG group, the difference was not statistically significant (P = .057). In histologic examination, the viability of bone in the VFF group, enchondral bone healing, and lamellar bone formation in the NVFG group were apparent., Conclusions: The results of this study suggest that NVFGs have a greater bone resorption rate than do VFFs. Furthermore, the histomorphometric results imply that reconstruction of the mandibular defects with vascularized osteoperiosteal femur flaps will provide greater strength.

Objectives: To examine the ultrastructural effects of maternal deprivation during developmental periods of limbic-hypothalamo-pituitary-adrenal system on hippocampal dendritic structures in adult rats., Methods: The experiments were carried out with male and female Wistar rats in our department. The rats were mated and, after birth, the pups were divided into four groups. The first group (control group) pups remained undisturbed with their dam until postweaning day 22. Maternal deprived groups were separated from their dams for 24 hours at postnatal day 4, 9 and 18. The subjects were provided with food and water ad libitum until 3-months-of-age. At the third month, the rats were transcardially perfused, samples were taken from CA1 and CA3 regions of the hippocampus. Tissues were prepared for electron microscopy., Results: When the data were analyzed, there were no differences between male and female rats in both ultrastructure and semiquantitative analysis of axodendritic synapses. The ultrastructure of Group 1 was seen as normal while in the second Group some neurons nuclear envelope made deep invagination into the nucleus. Additionally, axodendritic synapses were found normal. In Group 3, micrographs and axodendritic synapses were showed normal structure. However, in Group 4 in some neurons invaginations were seen similar to Group 2. Axodendritic synapses were found to be normal., Conclusion: These experiments establish that MD in rats produces slight ultrastructural changes and decreases the number of synapses in CA1 and CA3 subregions of the hippocampus.