Effects of Oxytocin on Glutamate Mediated Neurotoxicity in Neuroblastoma Cell Culture.

Introduction: We aimed to investigate the effects of oxytocin on neurite growth, cell viability, cell proliferation and apoptosis to demonstrate its neuroprotective effect on glutamate induced neurotoxicity in human neuroblastoma SH-SY5Y cell culture. Method: The effect of oxytocin on the toxic effects of glutamate in human neuroblastoma SH-SY5Y cell line with the Neurotoxicity Screening Test (NTT), apoptotic effects by Terminal Transferase dUTP Nick End Labeling (TUNEL) method and cell viability test by 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) method. In the NTT test; Neurotoxicity was induced by adding glutamate at a concentration of 32 µM to the cell culture. Oxytocin was added at 1, 3, 10, 30 and 100 µM concentrations and its effect on neurite elongation was investigated. It was demonstrated by TUNEL method that application of glutamate caused apoptosis. Afterwards, when glutamate and different doses of oxytocin were given, antiapoptotic effect was evaluated with the apoptotic index. Results: Glutamate was found to have a dose-dependent neurotoxic effect and reduced neurite elongation by 50% at a concentration of 32 µM. It was shown that the inhibition of neurite elongation caused by glutamate decreased in a dose-dependent manner by applying oxytocin. Especially oxytocin was found to significantly reduce neurite inhibition and show a neuroprotective effect starting from 10 µM concentrations. The concentration at which glutamate reduces cell proliferation by 50% was determined as 54 µM in MTT. Subsequently, it was observed that the adverse effect of glutamate on cell proliferation significantly decreased with oxytocin administration, depending on the dose. Conclusion: It was found that different concentrations of glutamate have a significant toxic effect on cell proliferation and viability, glutamate inhibits neurite elongation in a dose-dependent manner; oxytocin reduces neurite inhibition caused by glutamate, has a neuroprotective effect, increases cell viability and has antiapoptotic effects.

Histological and electroencephalographic demonstration of probiotic effect for reduce of oxidative stress and apoptosis in experimental traumatic brain injury.

BACKGROUND: The gut microbiota modulates nervous system function. In the literature, it has been shown that this modula-tion is used in many nervous system injuries through oxidative stress (OS) and apoptosis mechanisms. In this study, it was aimed to investigate the neuroprotective effects of probiotic (PB) treatment in a rat traumatic brain injury (TBI) model with histological and electroencephalographic (EEG) data. METHODS: Forty male Wistar albino rats were divided into four groups. Group 1 was the control group (CONTROL, n=10) and no trauma was applied. Group 2 was the trauma group with the weight-drop technique (TBH, n=10). Group 3 was the sham group (SHAM), (TBH+sterile saline [SS], n=10) rats were given 500 µL of SS per day by oral gavage. Group 4 was the PB treatment group, (TBH+PB, n=10) rats were treated daily for 7 days with 500 µL of PB oral gavage. Brain samples were collected 7 days after trauma. Histopathological evaluation of brain samples was done with HE. OS with Endothelial nitric oxide synthase, vascularization with Vas-cular Endothelial Growth Factor, gliosis with S100, and apoptosis with caspase 3 were evaluated immunohistochemically. Apoptotic index was determined with TUNEL. In addition, EEG and somatosensory evoked potential (SEP) recording findings were compared. RESULTS: It was determined by HE staining that there was a significant (P<0.001) damage in the TBI and sham groups compared to the control group. It was found that PB treatment provided a significant (P<0.01) improvement in the damage created. While OS (P<0.01), gliosis (P<0.01), and apoptosis (P<0.05) decreased with PB treatment, angiogenesis (P<0.01) increased. In support of these findings, in the software-mediated EEG and SUP examination; Delta wave power and theta/alpha ratio increased with TBI and de-creased with PB treatment. CONCLUSION: The results showed that PB treatment provided a significant improvement in rats by reducing OS, apoptosis, and gliosis and increasing vascularity. To the best of our knowledge in the literature, it was shown for the 1st time that histological results for the treatment of PB were supported by software-mediated EEG and SEP analysis.

The Condylar Effects of Mesenchymal Stem Cells, Low-Level Laser Therapy and Grape Seed Extract on Functional Mandibular Advancement of the Rat Mandible.

Objective: Functional treatment of Class II malocclusion is expected to lead to adaptation in the condyle. This study aimed to evaluate the effects of adipose tissue-derived mesenchymal stem cells (ADMSCs), low-level laser therapy (LLLT), and grape-seed extract (GSE) on condylar growth after functional mandibular advancement. Methods: Forty-five rats were randomly divided into 8 groups. Functional appliances were applied to all groups (n=6) except the control group (n=3). One group was treated with appliances only; the other six groups received various combinations of ADMSCs, LLLT, and GSE. Analyses for new osteoblasts and new bone formation, vascular endothelial growth factor, and Type II collagen were performed on condylar tissues, after an experimental period of four weeks. The quantitative data obtained from the results of the experiments were evaluated by H-score and analyzed using One-Way ANOVA by Tukey-Kramer multiple comparisons test (p≤0.05). Results: Levels of all investigated parameters increased in all groups (p≤0.05). The highest increases were achieved by a combined application of functional appliance, ADMSCs, LLLT and GSE (p≤0.05). Single LLLT administrations or single GSE applications did not create a statistical difference from appliance alone (p>0.05). A positive effect of ADMSCs or LLLT on osteoblast formation, neovascularization, and Type II collagen level was apparent (p≤0.05), however, neither affected new bone formation (p>0.05). Conclusion: This study shows that ADMSCs with LLLT and GSE applications provide differing levels of new osteoblast and bone formation, new vascular formation, and Type II collagen formation in rat condyles after functional mandibular advancement.

The Effect of Centrifuge Duration on Fat Graft Survival.

Background Although fat grafts are widely used for reconstruction and aesthetic purposes, their survival rates differ significantly. Centrifugation is one of the methods used to increase the survival of fat grafts. However, experimental studies examining the long-term outcomes of centrifugation duration are currently limited. Thus, in the present study, the effects of centrifugation duration on the survival of fat grafts were assessed using an animal model. Methods Thirty Sprague Dawley rats were included in the study and fat grafts were obtained from each specimen by excision from inguinal fat pads. Preparation protocols were administered as an en-bloc fat graft in Group 1, minced fat graft in Group 2, and fat graft centrifuged at 1,054 ×g for 2 minutes, 3 minutes, and 4 minutes in Group 3, 4, and 5, respectively. After 12 weeks of follow-up, grafts were harvested and were subjected to histopathological evaluation based on an established scoring system. Results En-block fat grafts were associated with necrosis, fibrosis, inflammation, vacuole formation, and alterations in adipocyte morphology. Among the three centrifugation groups, Group 3 demonstrated the best adipocyte viability and vascularity. However, graft weights decreased in all experimental groups. Conclusion The centrifugation process may have positive effects on adipocyte survival by means of purifying the fat graft and increasing adipocyte concentration. When the centrifugal durations were compared, 3-minute centrifuge yielded the most favorable results.

Efficacy of stromal vascular fraction and enzyme-free mechanical isolation therapy in experimental full thickness burn wounds.

BACKGROUND: Autologous cell suspensions obtained by a stromal vascular fraction (SVF) and enzyme-free mechanical isolation (EMI) are an alternative in the treatment of burn wounds. In this study, we aimed to investigate the effect of autologous cell suspensions obtained by SVF and EMI on full-thickness skin burn wounds. METHODS: A total of 45 male Sprague-Dawley rats were divided into three groups, SVF group, EMI group, and SVF + EMI group. The groups were also classified as the first, second, and third week of the burn to reveal the effect of the treatment on the burn in the early, middle, and late stages. For treatment, 0.2 ml SVF or 0.2 ml EMI was injected subcutaneously into the burn lesions of the subjects. Histopathological examination was performed on the burn wounds taken at the end of the experiment, and Ki67, CD44, CD73, CD90, and CK17 expressions were evaluated. RESULTS: Histological examination revealed that there was no improvement in the control samples, but the skin was multicellular, vascularization was present. Histologic scores in all groups was significantly better than control, and SVF + EMI was the best group in terms of recovery (p < 0.05). Ki67, CK17, CD44, CD73, and CD90 expressions were significantly higher in the treatment groups compared to the control (p < 0.05). CONCLUSION: We found in our study that both applications significantly increased the healing of the burn wound. Moreover, SVF + EMI application provided more improvement than SVF or EMI alone.

Adipose-Derived Mesenchymal Stem Cell Transplantation in Chemotherapy-Induced Premature Ovarian Insufficiency: the Role of Connexin and Pannexin.

In women undergoing chemotherapy, it is inevitable that infertility risk will increase because of impaired reproductive functions. Premature ovarian insufficiency (POI), which occurs as a devastating result of chemotherapy, is the complete depletion or dysfunction of ovarian follicles. Adipose-derived mesenchymal stem cells (ADMSCs) transplantation is among the alternative treatment methods for POI, which currently do not have an effective treatment method. Apoptosis of granulosa cells in POI is seen as the main mechanism of the disease. It is also reported that in addition to molecules directly associated with apoptosis, connexins, and pannexins are also potential effector molecules in apoptosis. The roles of these molecules in POI, which are known to play a role in many important mechanisms in the ovary, are unknown. In this study, it was aimed to analyze the expressions of Connexin43 and Pannexin1, which are thought to be effective in the formation of POI, and to show the relationship between the antiapoptotic effects of ADMSCs transplantation and these molecules in POI. For this purpose, Caspase3, Connexin43, Pannexin1 proteins, and mRNA expressions were analyzed by immunohistochemistry and RT-qPCR, and AMH levels were measured by ELISA. It was determined that Pannexin1, Caspase3 proteins, and mRNA levels increased in the POI, while Pannexin1 and Caspase3 expressions decreased in the ADMSCs treated group. While Connexin43 level decreased in POI, Connexin43 protein and mRNA levels increased in ADMSCs group. Consequently, this study demonstrated for the first time that Connexin43 and Pannexin1 were associated with apoptosis in POI. In addition, it was revealed that ADMSCs transplantation could produce antiapoptotic effects by modulating these molecules.

The relationship of 4-vinylcyclohexene diepoxide toxicity with cell death, oxidative stress, and gap junctions in female rat ovaries.

PURPOSE: It was aimed to investigate the damage caused by VCD toxicity in the ovary, which women working in the industrial field are frequently exposed to, and to show the relationship between gap junction protein, oxidative stress, and apoptosis, which is thought to be effective in the emergence of this damage. METHODS: Rats were divided into three groups as control, sham, and VCD. Histological stainings were performed for histopathological evaluations in ovary. Serum AMH level was measured with the ELISA. Then, iNOS, caspase 3, connexin 43 protein, and mRNA expression levels were analyzed by immunohistochemistry and RT-qPCR methods. RESULTS: As a result of the analyses, different amounts of degenerations such as hemorrhage, vacuolization, and fibrosis were observed in the ovary. VCD group AMH level decreased compared to control. In VCD group, iNOS and caspase 3 expressions increased, while connexin 43 expression decreased. CONCLUSIONS: It was shown that VCD caused damage to all ovarian tissue. Also, it was revealed for the first time that VCD triggered apoptosis by increasing oxidative stress in the ovary and suppressed connexin 43 which was also effective in the survival of granulosa cells. The devastating effect of exposure to occupational chemicals such as VCD on fertility was demonstrated in this study.

Therapeutic effects of Lacosamide in a rat model of traumatic brain injury: A histological, biochemical and electroencephalography monitoring study.

OBJECTIVE: Traumatic Brain Injury (TBI) is a major cause of death and disability worldwide, especially in children and young adults. TBI can be classified based on severity, mechanism or other features. Inflammation, apoptosis, oxidative stress, and ischemia are some of the important pathophys-iological mechanisms underlying neuronal loss after TBI. Lacosamide (LCM) is an anticonvulsant compound approved for the adjunctive treatment of partial-onset seizures and neuropathic pain. This study aimed to investigate possible neuroprotective effects of LCM in a rat model of TBI. MATERIAL AND METHODS: Twenty-eight adult male, Wistar albino rats were used. The rats were divided into 4 groups. Group 1 was the control group (n=7). Group 2 was the trauma group (n=7) where rats were treated with 100 mg/kg saline intraperitoneally (IP) twice a day. Groups 3 and 4, rats were treated with 6 (group 3, n=7) or 20 (group 4, n=7) mg/kg Lacosamide IP twice a day. For each group, brain samples were collected 72 hours after injury. Brain samples and blood were evaluated with histopathological and biochemical methods. In addition, electroencephalograpy monitoring results were compared. RESULTS: The immunoreactivity of both iNOS and eNOS (oxidative stress markers) were decreased with LCM treatment compared to trauma group. The results were statistically significant (***P<0.001). The treatments of low (56,17±9,69) and high-dose LCM (43,91±9,09) were decreased the distribution of HIF-1α compared to trauma group (P<0.01). The number of apoptotic cells were decreased with LCM treatment the difference between the trauma group and 20mg/kg LCM treated group (9,55±1,02) was statistically significant (***P<0.001). Malondialdehyde level was reduced with LCM treatment. MDA level was significantly higher in trauma group compared to LCM treated groups (***P<0.001). The level of Superoxide dismutase in the trauma group was 1,86 U/ml, whereas it was 36,85 U/ml in 20mg/kg LCM treated group (***P<0.001). Delta strength of EEG in 20mg/kg LCM treated group were similar to control group values after LCM treatment. CONCLUSION: No existing study has produced results suggesting that different doses of LCM has therapeutic effect against TBI, using EEG recording in addition to histological and biochemical evaluations in rats.

Effects of oxidative stress and apoptosis on vascularity and viability of perforator flaps.

We investigated lateral thoracic and posterior thigh perforator flaps for viability, vascularization, perfusion and apoptosis in a rat model. Wistar albino rats were divided into six groups: lateral thoracic artery perforator flap (LTPF) sham, 3 × 2 cm2 LTPF, 3 × 6 cm2 LTPF, posterior thigh perforator flap (PTPF) sham, 3 × 2 cm2 PTPF, and 3 × 6 cm2 PTPF. Flap viability was determined on postoperative days 1 and 7. On day 7, flaps were photographed and their viability was measured using two-dimensional planimeter paper. Tissue samples were harvested for examination by histology and immunohistochemistry. Viability differences were statistically significant. Epithelial thickness, vascularity and number of fibroblasts were reduced in the 3 × 6 cm2 groups. Neovascularization and apoptosis based on molecular tests were not significantly different among groups. Flap size and location are important factors for closure of surgical or traumatic defects. We suggest that for clinical application, wound complications will occur less frequently with perforators that nourish large areas of flaps.

Investigation of the effects of bisphenol-A exposure on lymphoid system in prenatal stage.

Bisphenol-A (BPA) used in the production of plastic materials is a temperature-soluble agent. It also has a steroid hormone-like activity; therefore, it poses a danger to human health. In our study, we aimed to investigate the effects of BPA on lymph node and spleen in male rats exposed to this agent during prenatal stage. The pregnant female rats were divided into four groups: control, sham, low dose (300 µg/kg BPA), and high dose (900 µg/kg BPA). BPA was dissolved in 1 mL of corn oil and administered to the pregnant rats every day during pregnancy. On the 21st and 45th day after the birth, male rats' lymph node and spleen samples were taken and histopathological examination was performed. Samples were stained with hematoxylin and eosin to determine the general histological appearance, and with CD3 and CD20 immunohistochemically. The results of staining were evaluated by H-score, and statistical analysis was performed. In the samples, BPA applications were not found to cause significant tissue damage. But there was a significant decrease in the immunoreactivities of CD3 and CD20 after BPA applications in both 21st and 45th day samples. After high dose BPA administration, decreased CD3 immunoreactivity was statistically significant. It is thought that BPA does not cause histologically significant tissue damage, but it may impair organ function at cellular level. The investigation of molecules involved in organ function will be useful in revealing the mechanisms that will cause dysfunction.

Adjuvant effects of chemotherapeutics and Metformin on MFE-319 endometrial carcinoma cell line.

We aimed to investigate the cytotoxicity of Metformin, Cisplatin, and Paclitaxel on MFE-319 endometrial carcinoma cell line using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and immunocytochemistry assays. Half maximal inhibitory concentration (IC50) doses of three drugs alone and in the dual combinations were applied to the cells. Immunocytochemical method was performed for the cell survival and for phosphatidylinositol 3-kinase (PI3K), phosphorylated extracellular regulated kinases (pErk)-1∕2, Akt-1, phosphorylated Akt (pAkt)-1∕2∕3 cell growth markers and angiogenic vascular endothelial growth factor (VEGF). Immunoreactivities were evaluated using H-score and analyzed using the one-way analysis of variance (ANOVA) test for statistics. It was found that these drugs caused a decrease in the immunoreactivities of these markers. Particularly, dual combination of Paclitaxel and Cisplatin decreased the immunoreactivities of PI3K, pErk-1∕2, Akt-1, and pAkt-1∕2∕3. Cisplatin and Paclitaxel were more effective than Metformin; on the other hand, Metformin has been shown to enhance the efficacy of these two drugs. In vitro or in vivo further studies are needed to investigate the efficacy of these three drugs via PI3K∕Akt signal pathway.

Effect of Different Dentin Conditioning Agents on Growth Factor Release, Mesenchymal Stem Cell Attachment and Morphology.

INTRODUCTION: EDTA has been considered the gold standard in regenerative endodontic treatments. The aim of this study was to evaluate the effects of different dentin conditioning agents other than EDTA on released growth factors, mesenchymal stem cell attachment, and morphology. METHODS: Transforming growth factor beta 1, vascular endothelial growth factor, bone morphogenetic protein 2, and fibroblast growth factor 2 release from prepared dentin discs conditioned with 17% EDTA, 10% citric acid, 1% phytic acid (IP6), or 37% phosphoric acid were quantified using the enzyme-linked immunosorbent assay after final irrigation and after 3 days of adipose-derived mesenchymal stem cell (adMSC) seeding. Forty root fragments were prepared from extracted single-rooted teeth. The morphology and attachment of adMSCs on the conditioned root fragments were observed using a scanning electron microscope. Data for growth factor quantification were analyzed using 1-way analysis. RESULTS: The highest transforming growth factor beta 1 release was observed after citric acid treatment followed by phosphoric acid; there was no significant difference between them, but compared with EDTA and 1% IP6, there were significant differences observed. The enzyme-linked immunosorbent assay detected a very minor exposure of vascular endothelial growth factor and fibroblast growth factor 2 after dentin conditioning, but there were no significant differences between the groups. The greatest bone morphogenetic protein 2 release was observed in the 1% IP6 group, but there were no significant differences between the groups. Three days of adMSC seeding after dentin conditioning has made a dramatic increase in all of the growth factors, and phosphoric acid appeared to be the most effective agent with significant differences compared with the remaining groups. Scanning electron microscopic observations showed that none of the conditioning solutions had an adverse effect on stem cell proliferation and attachment to root dentin. Different cell morphologies like round, oblong, flat, and well-attached cells with developed filopodia were observed in the dentin-conditioned groups. CONCLUSIONS: Phosphoric acid conditioning could be useful and may have beneficial effects in regenerative endodontic treatments.

Inhibiting effect of oleocanthal on neuroblastoma cancer cell proliferation in culture.

We investigated the potential anticancer effects of oleocanthal (OC) on neuroblastoma cells. Cells were divided into four groups: group 1, neuroblastoma cells were treated with OC; group 2, neurons that differentiated from neuroblastoma cells were treated with phosphate-buffered saline(PBS); group 3, bone marrow derived neuronal (BMDN) cells that were differentiated from bone marrow derived mesenchymal stem cells (BMSCs) were treated with OC; group 4, BMDN cells that were differentiated from BMSCs were treated with PBS. Groups 2 and 4 were control groups. The effects of OC on cell viability, oxidative stress, neurite inhibition and apoptosis at IC50 dose were investigated using MTT analysis, i-NOS and e-NOS measurement, neurotoxicity screening test (NST) and TUNEL staining, respectively. MTT analysis demonstrated that cells were significantly less viable in group 1 than in group 3. i-NOS and e-NOS staining intensity was significantly greater in group 1 than in group 3. NST revealed that OC inhibited neurite growth in both neuroblastoma and BMND cells; inhibition was significantly less in group 3 than in group 1. Significantly more TUNEL labeled cells were found in group 1 than in group 3. We found that OC prevented growth and proliferation of neuroblastoma cells in culture by increasing oxidative stress and apoptosis. We also found that the cytotoxicity of OC is negligible in BMDN cells.

The investigation of immunomodulatory effects of adipose tissue mesenchymal stem cell educated macrophages on the CD4 T cells.

Mesenchymal stem cells (MSCs) are strong immunomodulatory cells investigated in numerous clinical studies on fatal pathologies, such as graft versus host disease and autoimmune diseases; e.g., systemic lupus erythematosus, Crohn's disease, and ulcerative colitis. Macrophages are one of the critical cells linking the innate and adaptive immune system, and it has been shown that MSCs can differentiate between pro-inflammatory M1 phenotype and anti-inflammatory M2 phenotype of macrophages. However, it has not yet been fully clarified whether these differentiated macrophages are functional. In this study, we compared the immunomodulatory effects on the CD4 T cells of M1, M2a and M2c macrophages with the macrophages that directly and indirectly cultured with MSCs. We analyzed the changes in CD14, CD64, CD80, CD163 and CD200R expression to evaluate macrophage phenotypes, and the changes in CD4, IFN-g, IL-4, IL-17a and FoxP3 expression to evaluate T helper subsets using the FACS method. The changes in IL-1b, IL-4, IL-10, IL-12p70, IL-17a and IFN-g in the media supernatants were analyzed using the Luminex method. We also performed WST-1 and Caspase-3 ELISA analyses to observe the proliferation and apoptosis status of the T cells. MSCs were found to differentiate macrophages into a distinctive phenotype, which was close to the M2c phenotype, but was not considered as an M2c cell due to the low expression of CD163, a characteristic marker for M2c. While MEM-D, MEM-ID and MSCs showed similar inhibitory effects on the Th2 and Th17 cells, the most significant increase in Treg cell frequencies was seen in MEM-D cells. Macrophages can alter their phenotypes and functions according to the stimuli from the environment. The fact that macrophages educated with MSCs suppressed the production of all the cytokines we evaluated even after the removal of MSCs suggests that these cells may be differentiated by MSCs into a suppressive macrophage subgroup. However, the Treg cell activation caused by direct interactions between MSCs and macrophage cells may be the most prominent observation of this study compared to previous work. As a result, according to our data, the interactions between MSCs and macrophages may lead to differentiation of macrophage cells into an immunosuppressive phenotype, and these macrophages may suppress the T lymphocyte subgroups at least as effectively as MSCs. However, our data obtained from in vitro experiments should be supported by future in vivo studies.

Effects of boric acid on bone formation after maxillary sinus floor augmentation in rabbits.

PURPOSE: Augmentation of the maxillary sinus floor with bone grafting is commonly used for successful treatment of edentulous posterior maxilla with dental implants, and it is essential to maintain good bone volume and quality for long-term success of dental implants. The aim of this experimental study was to investigate the local and systemic effects of boric acid on new bone formation after maxillary sinus floor augmentation (MSFA). MATERIALS AND METHODS: Twenty-four male, New Zealand rabbits were randomly divided into three groups with eight rabbits each, and bilateral MSFA was performed in each animal. An autogenous bone/xenograft mixture was used to augment the maxillary sinuses in each group. Group 1 was determined as control with no additional materials, whereas 3 mg/kg boric acid (BA) was added to the mixture in group 2, and 3 mg/kg boric acid solution added to drinking water daily in group 3. RESULTS: The animals were sacrificed and also histologic, histomorphometric, and immunnohistochemical analyses were performed at weeks 4 and 8. At week 4, bone regeneration was better in the local BA group than in the control and systemic BA groups (p < 0.05). However, no significant difference was found among the groups in terms of bone regeneration at the end of week 8 (p > 0.05). CONCLUSION: Significant higher new bone formation was revealed by BA at early healing especially with local application. BA may be a therapeutic option for improving the bone regeneration.

Histological investigations on thymus of male rats prenatally exposed to bisphenol A.

Bisphenol A is called as a endocrine-distrupting chemical because of the its steroid-like activity and it used in the construction of plastic containing materials. It is indicated that bisphenol A can pass the human serum, urine, follicular fluid, placenta and umblical cord as a result of the use of substances containing this agent. In this study, we aimed to investigate the effects of bisphenol A on the development of the thymus, a primary lymphoid organ which plays an important role in the specific immunity. The adult pregnant female rats were administered orally with bisphenol A (for 21 days) and postnatal thymus samples were obtained on day 21, 45 and 90 and were performed for histochemical and immunohistochemical staining for CD3, CD4, CD8 and CD79a and TUNEL assay for the apoptotic cells. Evaluation of all groups, CD3, CD4, CD8 and CD79a stainings were decreased in the experimental groups compared with control group. The apoptotic cells were determined in the all groups on day 90 as a result of the thymus involution. It is noted that there was not any histological and morphological damages in the rats prenatally exposed the bisphenol A. The effect of the bisphenol A is unknown in the future, but there is no problem in the adult rats.

Neuroprotective Effects of Oleocanthal, a Compound in Virgin Olive Oil, in a Rat Model of Traumatic Brain Injury.

AIM: To evaluate the neuroprotective effects of deocanthal OC in a rat model of traumatic brain injury (TBI). MATERIAL AND METHODS: Twenty-six adult male, Wistar albino rats were used. The rats were divided into 4 groups. Group 1 was the sham group (n=5). Group 2 was the trauma group (n=5) where rats were treated with 10 mg/kg saline intraperitoneally (IP) twice a day. Groups 3 and 4, rats were treated with 10 (group 3, n=8) or 30 (group 4, n=8) mg/kg OC IP twice a day. For each group, brain samples were collected 72 hours after injury. Brain samples and blood were evaluated with histopathological and biochemical methods. RESULTS: Histopathological evaluation revealed a significant difference between Group 2 and Group 4. Biochemical findings demonstrated that the oxidative stress index was highest in Group 2 and lowest in Group 4. CONCLUSION: OC has a protective effect on neural cells after TBI. This effect is achieved by reducing oxidative stress and apoptosis.

Effects of lacosamide "a novel antiepileptic drug" in the early stages of chicken embryo development.

INTRODUCTION: Antiepileptic drugs (AEDs) are teratogens and confer a risk of congenital malformation. The estimated prevalence of major congenital malformations such as cardiac defects, facial clefts, hypospadias, and neural tube defects in epileptic women is 4-10 %, which represents a two- to fourfold increase in pregnant women compared to the general population. However, there are no clear data for newer drugs. Lacosamide (LCM), a novel AED, is the first of the third-generation AEDs to be approved as adjunctive therapy for the treatment of partial-onset seizures. There are no data on the pharmacokinetics of LCM during pregnancy, and only some published data have reported on its effects during pregnancy. METHODS: In this study, three different doses of LCM (0.12, 0.5, and 1.60 mg in 0.18 mL) were applied under the embryonic disks of specific pathogen-free Leghorn chicken embryos after a 30-h incubation. Incubation was continued for 80 h, at which time all embryos were evaluated macroscopically and microscopically. RESULTS: There was growth retardation in all of the LCM-treated groups. Major malformations increased in a dose-dependent manner and were mostly observed in the supratherapeutic group. CONCLUSION: Based on our data, LCM may cause growth retardation or major congenital malformations. Nevertheless, more extensive investigations of its reliability are needed.

Matrix changes due to the toxic effects of metronidazole in intestinal tissue of fish (Onchorhynchus mykiss).

Metranidazole (MTZ) is an antibiotic used for parasitic infections in a number of species. Accumulation of this drug in the environment and its interaction with fish of economic value makes this drug particularly important. In the present study, we examined the histopathological effects of MTZ on the intestinal tissue of Oncorhynchus mykiss. The fish in aquarium were exposed to MTZ at doses of 5, 10, 20 mg/L for 2, 4 and 8 days. At the end of the experiments, macroscopic pathology or death were not observed at these doses. Histochemical staining with Haematoxylene-Eosin, Periodic Acid Schiff and Gomori Trichrome showed, depending on increased dose and prolonged duration, areas of necrosis, edema, inflammation, small tears at the tips of the villi and excretion with heterogenic distribution of the Goblet cells. Moreover, changes in the connective tissue of the intestines due to toxicity of MTZ and decreases in immunostaining of matrix proteins such as laminin and collagen IV, especially in the epithelium were observed. Findings of the present study would be useful to demonstrate the adverse effects of MTZ use, emphasizing the importance of the effect on fish which could be very important public health.