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EgeUn###, Neurodegenerative diseases affect millions of people. Major reasons behind the onset and progression of these diseases are still under investigation. Therefore, any approach that would treat/prevent progression is important. In this study, we aimed to investigate the potential protective effects of Psephellus pyrrhoblepharus (Boiss.) Wagenitz extracts in MPP+-induced dopaminergic cell damage and compare the effectiveness of different extracts (methanol:water (1:1), chloroform and n-hexane). The cells were pretreated with four different concentrations (10, 50, 100, and 200 mu g/ml) of methanol:water (1:1), chloroform and n-hexane extracts of P. pyrrhoblepharus following MPP+ treatment for 12 or 24 h. The changes in cell viability were determined using the MTT assay. Additionally, antioxidant activities and total phenolic/flavonoid contents of the extracts were determined with radical scavenging capacity, Folin-Ciocalteu and aluminum chloride assays, respectively. The extracts at selected concentrations were found to be protective in a dose-dependent manner at 12 and 24 h. Nevertheless, the methanol extract of the plant showed the highest protection both at 100 and 200 mu g/ml (115.13%+/- 3.98, 121.87%+/- 1.66; p < 0.05) against dopaminergic damage at 24 h. The results showed that selected concentrations were not toxic and did not affect cell proliferation rate. Besides, the chloroform extract was found to have higher antioxidant activity than the other extracts (p < 0.05). The total phenolic and total flavonoid contents were found consistent with antioxidant activities. Our findings support the neuroprotective and antioxidant potential of P. pyrrhoblepharus. However, further studies on identifying the presence of chemicals in P. pyrrhoblepharus extracts which are responsible for protection should be carried out to confirm their therapeutic potential.

Purpose: The present study was conducted to determine oxidative stress and cell viability after contact with resin composites polymerized for different times. Methods: Disk-shaped specimens of Admira Fusion. Ceram X One Universal. Solare x and Filtek Z550 (n = 12) were prepared, and two subgroups with polymerization times of 20 and 40 s were employed. The specimens were incubated with mouse fibroblast cells for 48 and 72 h, and changes in reactive oxygen species (ROS) production and cellular viability were determined by an assay with a cell-permeable fluorescent dye, 2',7'-dichlorodihydrofluorescein diacetate (H2DCF-DA), and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, respectively. Results: At 72 h, ROS production in the presence of Admira Fusion polymerized for 40 s was reduced relative to that in the presence of Admira Fusion polymerized for 20 s (P < 0.05). Cell viability was maximal in the Admira Fusion and Solare x groups and there was no difference relative to the control group at 48 h. Cell viability was higher in the Admira Fusion and Solare x groups polymerized for 40 s than for the same materials polymerized for 20 sat 72 h (P < 0.05). Conclusion: Extension of the polymerizaton lime has a material-specific effect and may be used as a strategy to increase the biocompability of resin composites.

Experimental animal study investigating the efficacy of C-terminal domain of tetanus toxin application as neuroprotective effects on rat brain in a model of spinal cord injury (SCI).The aim of the present study was to investigate the possible role of C-terminal domain of tetanus toxin (Hc-TeTx) on cell death mechanisms including apoptosis and autophagy following SCI.Traumatic SCI can lead to posttraumatic inflammation, oxidative stress, motor neuron apoptosis, necrosis, and autophagy of tissue. To promote and enhance recovery after SCI, recent development of devices and therapeutic interventions are needed.Twenty-eight adult rats were divided into four groups (n = 7 each) as follows: sham, trauma (SCI), SCI + Hc-TeTx, and SCI + methylprednisolone groups. The functional neurological deficits due to the SCI were assessed by behavioral analysis using the Basso, Beattie and Bresnahan (BBB) open-field locomotor test. The alterations in pro-/anti-apoptotic and autophagy related-protein levels were measured by Western blotting technique.In this study, Hc-TeTx promotes locomotor recovery and motor neuron survival of SCI rats. Hc-TeTx also decreased expression of bax, bad, bak, cleaved caspase-3, Ask1, and autophagy-related proteins including Atg5 and LC3II in brain. Our study provides an evidence that cell death mechanisms play critical roles in SCI and that the nontoxic peptides including Hc-TeTx may exert protective effect and decrease cell death following SCI.Our preliminary findings suggest a possible therapeutic agent to improve survival after spinal cord trauma, but further analysis are still needed to evaluate the difference between acute and chronic injuries.N/A.

Neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, and Huntington's disease affect millions of people in the world. Thus several new approaches to treat brain disorders are under development. The aim of the present study is to synthesize potential neuroprotective heterocyclic compounds based on pyrazolopyridine derivatives and then to evaluate their effects in MPP+-induced neurodegeneration in human neuroblastoma cell line (SH-SY5Y cells). The effects of the compounds on cell viability were measured by MTT assay and the changes in apoptosis-related proteins including bax, Bcl-2, Bcl-xl and caspase-3 were investigated by western blot technique. Based on the cell viability results obtained by MTT assay, the percentage of neuroprotection-induced by compounds against MPP+-induced neurotoxicity in SH-SY5Y cells was between 20% and 30% at 5 μM concentrations of all synthesized compounds. Moreover, the downregulation in pro-apoptotic proteins including bax and caspase-3 were found following the novel synthesized compounds treatments and these effects were observed in a dose-dependent manner. Our results provide an evidence that these heterocyclic compounds based on pyrazolopyridine derivatives may have a role on dopaminergic neuroprotection via antiapoptotic pathways.

The loss of neurons is strongly correlated with aging and aging-associated disorders. in this study, cell viability assays and mitochondrial function were performed to evaluate the effect of new spiro-pyrazole derivatives, prepared from aldehydes and 3-amino-1-phenyl-2-pyrazolin-5-one, on neuroprotection in an in vitro model of dopaminergic cell death induced by 1-methyl-4-phenylpyridinium (MPP+). The percentages of neuroprotection by derivatives were found between 21.26% and 52.67% at selected concentrations (10-50 mu M) with compound 4d exerting the best neuroprotective effect. The results show that the studied spiropyrazolones perform important roles in dopaminergic neuroprotection and can be used for potential new therapies in the treatment of neurodegenerative disorders including Parkinson's disease. (C) 2020 Elsevier Masson SAS. All rights reserved., University of AveiroEuropean Commission; Fundacao para a Ciencia e a Tecnologia (FCT, Portugal)/Ministerio da Ciencia, Tecnologia e Ensino Superior (MCTES, Portugal) [UIDB/50006/2020]; FEDER, within the PT2020 Partnership Agreement; Sultan Moulay Slimane University of Beni-Mellal (Morocco); National Center for Scientific and Technical Research (CNRST) -Morocco; FCT-CNRST (Morocco); FCTPortuguese Foundation for Science and TechnologyEuropean Commission [PD/BD/52531/2014], Thanks are due to the University of Aveiro and Fundacao para a Ciencia e a Tecnologia (FCT, Portugal)/Ministerio da Ciencia, Tecnologia e Ensino Superior (MCTES, Portugal) for the financial support for the LAQV-REQUIMTE (UIDB/50006/2020), through national founds and, where applicable, co-financed by the FEDER, within the PT2020 Partnership Agreement, and to the Portuguese NMR Network. The authors also thank the Sultan Moulay Slimane University of Beni-Mellal (Morocco) and the National Center for Scientific and Technical Research (CNRST) -Morocco for the funding the characterization analysis and to the Transnational cooperation programs, FCT-CNRST (Morocco), for financial assistance (2019-2020). C.M.B. Neves is also grateful to FCT for her Ph.D. Grant with the reference PD/BD/52531/2014. Authors also thank the Pharmaceutical Sciences Research Centre (FABAL, Ege University, Faculty of Pharmacy, Turkey) for equipmental support.

Increased concentration of manganese (Mn) in the brain is known to be associated with excitotoxicity and neuroinflammation. Vinpocetine, an alkaloid derived from the plant Vinca minor L., basically shows its effect via phosphodiesterase inhibition and voltage-dependent Na+ channels. Vasoactive intestinal peptide (VIP) has gastrointestinal, vasomotor, muscular, and neuroprotective effects. The aim of this study was to examine the potential protective effects of vinpocetine and VIP against Mn toxicity in NE-4C neural stem cells (NSCs). VIP treatment at 1 μM and vinpocetine treatment at 2 μM concentrations were sufficient to yield maximum protection, and these concentrations were adopted in the following experiments. In this study, Mn treatment significantly increased lactate dehydrogenase (LDH) leakage, reactive oxygen species (ROS) production, and triggered cell death in NE-4C cultures. However, significant reduction in LDH release was observed following vinpocetine or VIP treatments when compared with control. Similar to these findings, vinpocetine or VIP treatments significantly reduced membrane degradation induced by Mn (p

WOS: 000474396700008, PubMed: 30415440, ObjectiveThe aim of this in vitro study is to evaluate the effects of resveratrol (RES) addition on the cytotoxicity and microtensile bond strength (mu TBS) of different adhesives.Materials and methodsFive self-etching adhesives (G-aenial Bond-GC, Optibond All in One-Kerr, Gluma Self Etch-Kulzer, Clearfil S-3 Bond-Kuraray, and Nova Compo-B Plus-Imicryl) were tested. They were applied to L-929 cell culture by the extract method. In the test groups, 0.5 mu M RES (Sigma-Aldrich) was added into the medium. Cell viability was assessed by MTT assay after 24h. Human extracted third molars were used for mu TBS test (n=7). The adhesives with or without 0.5 mu M RES addition were applied on dentin surfaces. A composite build-up was constructed. Then, the specimens were sectioned into multiple beams with the non-trimming version of the microtensile test and subjected to microtensile forces. Statistical analysis was performed using ANOVA and post hoc Tukey test (p?0.05).ResultsThe extracts of all adhesives decreased the cell viability. However, RES addition increased the cell viability in all groups (p?0.05). RES addition did not cause any decrease in mu TBS values of the adhesives compared to baseline. Optibond All in One showed the highest mu TBS after RES addition. It was followed by Clerafil S-3 Bond and Nova Compo-B Plus. No difference was determined between the Optibond All in One and Clearfil S-3 Bond. There was difference between Optibond All in One and Nova Compo-B Plus (p?0.05).ConclusionRES addition may improve the biocompatibility without causing negative influence on mu TBS of the adhesives.Clinical relevanceRES addition has clinical applicable potential to overcome the adverse biocompatibility of adhesives.

Recently, nuclear translocation and stability of nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2) have gained increasing attention in the prevention of oxidative stress. The present study was aimed to evaluate the regulatory role of glycogen synthase kinase-3&beta, (GSK-3&beta, ) inhibition by tideglusib through the Nrf2 pathway in a cellular damage model. Gene silencing (siRNA-mediated) was performed to examine the responses of Nrf2-target genes (i.e., heme oxygenase-1, NAD(P)H:quinone oxidoreductase1) to siRNA depletion of Nrf2 in MPP+-induced dopaminergic cell death. Nrf2 and its downstream regulated genes/proteins were analyzed using Real-time PCR and Western Blotting techniques, respectively. Moreover, free radical production, the changes in mitochondrial membrane potential, total glutathione, and glutathione-S-transferase were examined. The possible contribution of peroxisome proliferator-activated receptor gamma (PPAR&gamma, ) to tideglusib-mediated neuroprotection was evaluated. The number of viable cells and mitochondrial membrane potential were increased following GSK-3&beta, enzyme inhibition against MPP+. HO-1, NQO1 mRNA/protein expressions and Nrf2 nuclear translocation significantly triggered by tideglusib. Moreover, the neuroprotection by tideglusib was not observed in the presence of siRNA Nrf2. Our study supports the idea that GSK-3&beta, enzyme inhibition may modulate the Nrf2/ARE pathway in cellular damage and the inhibitory role of tideglusib on GSK-3&beta, along with PPAR&gamma, activation may be responsible for neuroprotection.

Oxidative stress and apoptosis are both associated with various acute and chronic disorders. Thus, the aim of the present study is to synthesize imidazo[2,1-c][1,2,4]triazines derivatives and to evaluate their effects in H2O2-induced oxidative stress in human neuroblastoma cell line (SH-SY5Y cells). The effects of the compounds on cell viability were measured by MTT assay and the changes in stress and apoptosis-related proteins were investigated by PathScan® Stress and Apoptosis Signaling Antibody Array kit and Western Blot technique. In particular, four compounds were found to protect SH-SY5Y cells from H2O2-induced toxicity by increasing Bcl-2/Bax ratio, regulating PI3-K/Akt cascade and inhibiting the ERK pathway.

Objective: In Psephellus and Centaurea genuses a lot of species have important pharmacological activities like antipyretic, antiinflamatory, diuretic, cytotoxic etc. In Psephellus genus the endemic plant Psephellus pyrrhoblepharus (Boiss.) Wagenitz (Centaurea pyrrhoblephara) was collected from Elazığ, Turkey. Recently interest in use of plant-derived compounds for therapeutic purposes in cancer is increasing day by day. Liver cancer which is one of the cancer types is affecting millions of people all over the world. So any approach to treat liver cancer is extremely valuable. Materials and methods: We have purposed to determine the cytotoxic activity of methanol:water (1:1), n-hexane and chloroform extracts of the aerial parts of P. pyrrhoblepharus using human liver cancer cell (HepG2) by utilizing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay in different concentrations (50, 100 ,200 µg/ml) and time points (6, 12, 24 h). Results: Regarding the cytotoxicity, cell viability was seen as decreased following extract exposures at three different time points. The highest cytotoxic activity was observed in 100 µg/ml concentrations of chloroform extract at 24 h. At 12 h chloroform extract of plant showed the highest cytotoxic activity in 200 µg/ml concentrations when compared to methanol:water and n-hexane extracts. Conclusion: It can be suggested that extracts of P. pyrrhoblepharus show dose-dependent and time-dependent cytotoxic effects in HepG2 carsinoma cells. In general, the results provide information about the threshold concentrations of P. pyrrhoblepharus extracts that might be used in different applications without the risk of toxicity., Amaç: Birçok Psephellus ve Centaurea türü antipiretik, antiinflamatuar, diüretik ve sitotoksik gibi önemli farmakolojik aktivitelere sahiptir. Endemik bir bitki olan Psephellus pyrrhoblepharus (Boiss.) Wagenitz, Elazığ, Türkiye’den toplanmıştır. Son günlerde kanserde terapötik amaçlar için bitki kaynaklı bileşiklerin kullanımına olan ilgi artmaktadır. Bir kanser türü olan karaciğer kanseri de tüm dünyada milyonlarca insanı etkilemekteyken, karaciğer kanserini tedavi etmek için herhangi bir yaklaşım son derece değerlidir. Bu çalışmada, insan karaciğer kanseri hücresi (HepG2) kullanılarak P. pyrrhoblepharus bitkisinin topraküstü kısımlarından hazırlanmış olan metanol:su (1:1), kloroform ve n-hekzan ekstrelerinin sitotoksik potansiyelini değerlendirilmesi amaçlanmıştır. Gereç ve yöntemler: 3-(4,5-dimetiltiyazol-2-il)-2,5-difenltetrazolyumbromit (MTT) metodu kullanılarak, insan karaciğer kanser hücre hatlarında (HepG2) farklı konsantrasyonlarda (50, 100, 200 ug/ml) MTT, farklı zaman noktalarında (6, 12, 24 s) çalışmalar gerçekleştirildi. Bulgular: Sitotoksisite ile ilgili olarak, farklı zaman noktalarında ekstrelerin maruziyetlerinin ardından hücre canlılığının azaldığı gözlenmiştir. Bununla birlikte, en yüksek sitotoksik aktivite, 24 saatte 100 ug/ml konsantrasyondaki kloroform ekstresinde gözlenmiştir. Bitkinin kloroform ekstresi, metanol:su ve n-hekzan ekstreleri ile karşılaştırıldığında 12 saatte 200 ug/ml konsantrasyonda en yüksek sitotoksik etkinliği göstermiştir. Sonuç: P. pyrrhoblepharus ekstrelerinin HepG2 karsinoma hücrelerinde doza ve zamana bağlı bir şekilde sitotoksik etki gösterdiği ileri sürülebilir. Genel olarak elde edilen sonuçlar, toksisite tehlikesi olmadan farklı uygulamalarda kullanılabilecek P. pyrhoblepharus ekstrelerinin eşik konsantrasyonları hakkında bilgi sağlamıştır.

Spinal cord injury (SCI) leads to vascular damage and disruption of blood-spinal cord barrier which participates in secondary nerve injury. Epidermal growth factor (EGF) is an endogenous protein which regulates cell proliferation, growth and differention. Previous studies reported that EGF exerts neuroprotective effect in spinal cord after SCI. However, the molecular mechanisms underlying EGF-mediated protection in different regions of nervous system have not shown yet. In this study, we aimed to examine possible anti-apoptotic and protective roles of EGF not only in spinal cord but also in brain following SCI. Twenty-eight adult rats were divided into four groups of seven animals each as follows: sham, trauma (SCI), SCI + EGF and SCI + methylprednisolone (MP) groups. The functional neurological deficits due to the SCI were assessed by behavioral analysis using the Basso, Beattie and Bresnahan (BBB) open-field locomotor test. The alterations in pro-/anti-apoptotic protein levels and antioxidant enzyme activities were measured in spinal cord and frontal cortex. In our study, EGF promoted locomotor recovery and motor neuron survival of SCI rats. EGF treatment significantly decreased Bax and increased Bcl-2 protein expressions both in spinal cord and brain when compared to SCI group. Moreover, antioxidant enzyme activities including catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) were increased following EGF treatment similar to MP treatment. Our experiment also suggests that alteration of the ratio of Bcl-2 to Bax may result from decreased apoptosis following EGF treatment. As a conclusion, these results show, for the first time, that administration of EGF exerts its protection via regulating apoptotic and oxidative pathways in response to spinal cord injury in different regions of central nervous system.

Bir dental implant tedavisinin başarısında cerrahi teknik, implant materyali ve implant dizaynı gibi faktörlerin yanı sıra, implant materyalinin yüzey özelliklerinin de önemli olduğu bilinmektedir. Son yıllarda yapılan çalışmalar iyon implantasyonunun dental implantların yüzey modifikasyonu açısından iyi bir aday olduğunu göstermiştir._Bu çalışmada, MEVVA iyon implantasyon teknolojisi kullanılarak, metal+gaz hibrit iyon implantasyonu yapılmış yüzeylerin osteoblast hücre tutunmasına etkisi incelenmiştir. Bu amaçla, iyon implante edilecek materyal olarak son yıllarda dental implant materyali olarak en çok kullanılan titanyum (ISO 5832-2 Grade 4) seçilmiştir. Titanyum yüzeylere implante edilecek metal element olarak yüksek biyouyumluluğu ile bilinen zirkonyum ve zirkonyum ile eş zamanlı implante edilen gaz elementi olarakta titanyum yüzeylerin pasivasyonunu arttırmak için oksijen kullanılmıştır. Osteoblast hücre tutunmaları in vitro ortamda gerçekleştirilerek yüzey pürüzlülüğünün ve iyon implantasyon işleminin osteoblast hücre tutunmaları üzerine etkisi karşılaştırılmalı olarak incelenmiştir. Sonuç olarak pürüzlü ve iyon implante edilmiş yüzeylerde hücrelerin daha iyi tutunduğu görülmüştür, Achivement of a dental implant treatment depend on also surface properties of implant material as well as surgical method, implant material and implant desing, Studies have shown that ion implantation is a good candidate for modifying of dental implant surface._In this thesis, it was observed the effects on osteoblast attachment on modified surfaces with metal+gas hybrid ion implantation by using MEVVA ion implantation technology. For this aim, the material for ion implantation was chosen as titanium (ISO 5832-2 Grade 4) which is the most common material used in dentistry. Zirconium element known as biocompatible and oxygen to increase the passivation of titanium surfaces were used for creating ions simultaneously._ Attachment of osteoblast on samples was investigated in vitro for comparing the effects of the rougnesses and ion implantation surface modification on attachment of osteoblast. It is observed at the end of the in vitro osteoblast cell attachment is better on rough and ion implanted surfaces

Although hyperbaric oxygen (HBO) treatment following spinal cord injury (SCI) have been studied in terms of neurological function and tissue histology, there is a limited number studies on spinal cord tissue enzyme levels.The effect of HBO treatment in SCI was investigated by measuring superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), nitric oxide synthase (NOS) and nitric oxide (NO) activity in the injured tissue. SCI was induced by applying an aneurysm clip extradurally at the level of T9-T11 vertebrae. Preoperative HBO (preopHBO) treatment was applied for 5days and postoperative HBO (postopHBO) for 7days.In the preopHBO group, a significant decrease was observed in NOS and NO compared to the SCI group. There was a decrease in SOD, NOS and NO in the postopHBO group when compared to the SCI group. In the pre-postHBO group SOD, GPx, NOS and NO decreased significantly. There was a decrease in SOD in postopHBO compared to preopHBO. In the prepostopHBO, SOD decreased significantly compared to that in the preopHBO group. The prepostopHBO presented a significant decrease in GPx compared to postopHBO (p0.05 for all parameters). No significant difference was observed for catalase for all groups. Significant improvement was found in BBB scores for both postopHBO and prepostHBO groups when compared to the SCI group (p0.05).HBO treatment was found to be beneficial following SCI in terms of biochemical parameters and functional recovery in the postoperative period.

CART mRNA and peptides are highly expressed in the anatomical structures composing the hypothalamo-pituitary-adrenal (HPA) axis and sympatho-adrenal system. Anatomical and functional studies suggest that CART peptides may have a role in the regulation of the neuroendocrine and autonomic responses during stress. Our previous study showed that CART peptides increased significantly in the male hypothalamus and amygdala 10 min after the forced swim stress. The present study aimed to examine the effect of forced swim stress on CART peptide expression in selected brain regions, including those where CART peptide expression has not been reported before (frontal cortex, pons, medulla oblongata), as well as in endocrine glands related to stress in male Sprague Dawley rats. A total of 16 (n = 8) animals were used, including control groups. Rats were subjected to forced swim on two consecutive days, and sacrificed on the second day, 2 h after the termination of the stress procedure. Frontal cortex, pons, medulla oblongata, hypothalamus, pituitary and adrenal glands were dissected and homogenized. CART peptide expression in these tissues was measured by Western Blotting and six different CART peptide fragments were identified. Our results showed that forced swim stress elicited region-specific changes in CART peptide expression. CART was upregulated in the frontal cortex, hypothalamus, medulla oblongata and adrenal gland while there was no change in the pons and pituitary. Enhanced CART peptide fragments in these brain regions and adrenal glands may have a role in the regulation of the HPA and sympatho-adrenal axis activity during stress response.

We investigated functional recovery of the lower urinary system in rats with spinal cord injury after transplanting neuronal restricted precursors/glial restricted precursors or neural cells derived from bone marrow stromal cells into the injured area of the spinal cord.A total of 30 rats underwent experimentation in 4 groups, including group 1--sham operation, group 2--spinal cord injury plus neuronal restricted precursor/glial restricted precursor transplantation, group 3--spinal cord injury plus bone marrow stromal cell transplantation and group 4--spinal cord injury control. All rats in the 4 groups were investigated urodynamically and sacrificed on day 28 after transplantation. The cells transplanted into the injured spinal cord underwent histological investigation.Transplanted cells (neuronal and glial restricted precursors, and bone marrow stromal cells) were found to maintain a presence in the injured spinal cord area. Baseline pressure, maximum capacity, mean uninhibited contraction amplitude, mean voiding pressure, voided volume and post-void residual volume were significantly better in groups 2 and 3 than in group 4, while baseline pressure in group 2 was better than that in group 3. We found no significant difference among the groups according to mean uninhibited contraction frequency.Although neuronal/glial restricted precursor transplanted rats seemed to have more improvement, all rats in groups 2 and 3 showed some significant improvement in lower urinary system function. On the other hand, the level of this improvement was far from complete functional recovery.

Results of electrophysiological studies suggest a significant role of the lateral spinal nucleus (LSpN) in the transmission of nociceptive signals. In our study, the presence of Fos immunoreactivity and NADPH-diaphorase positivity was observed in the rat LSpN following noxious peripheral subcutaneous stimulation. Formalin-induced unilateral hindpaw stimulation in the rat caused bilateral NADPH-d reactivity and ipsilateral Fos expression in this nucleus. In the LSpN of the L3-L5 segments of stimulated rats, on average, 4.1 +/- 1.2 NADPH-d-positive, NADPH-d(+), 5.1+1.8 Fos-immunoreactive, Fos(+), and 3.0 +/- 1.1 double-labeled neurons per 25-mu m-thick section were found unilaterally. A close anatomical relationship between NADPH-d(+) processes and Fos(+) cell nuclei in the LSpN was also observed following noxious peripheral stimulation. These neuroanatomical findings support the hypothesis that the LSpN is involved in pain processing and suggest an important role of nitric oxide-mediated signal transduction in this nucleus.

WOS: 000378324000020, PubMed ID: 26981753, The aim of this study was to evaluate whether medium modification improves the odontogenic differentiation of human dental pulp stem cells (DPSC) in vitro and in vivo. DPSC isolated from human impacted third molar teeth were analysed for clusters of differentiation with flow cytometry. Odontogenic differentiation was stimulated by medium modification with the addition of bone morphogenetic protein 2 (BMP2). The expression of dentin sialophosphoprotein, dentin matrix protein 1, enamelysin/matrix metalloproteinase 20 and the phosphate-regulating gene with homologies to endopeptidases on the X chromosome of the cells were analysed with RT-PCR at 7, 14 and 21 days. Then, DPSC were transplanted on the back of immunocompromised mice via a hydroxyapatite tricalcium phosphate scaffold, and the structure of the formed tissue was investigated. The cells were identified as mesenchymal stem cells with a 98.3% CD73 and CD90 double-positive cell rate. The increase in mineralization capacity and expression of human enamel-dentin specific transcripts proportional to the culture period were determined after differentiation. Six weeks after transplantation, an osteo-dentin matrix was formed in the group in which odontogenic differentiation was stimulated, and the odontogenic characteristics of the matrix were confirmed by histological examination and RT-PCR analysis. Odontogenic differentiation of the isolated and characterized human DPSC was improved with medium modification by the addition of BMP2 in vitro and in vivo. The defined medium and applied technique have a potential use for forming reparative dentin in the future, but the effects of the method should be investigated in long-term studies., Ege University's Scientific Research FoundationEge University [2009-Dis-036]; Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK), This study was supported by Ege University's Scientific Research Foundation (2009-Dis-036). We also acknowledge The Scientific and Technological Research Council of Turkey (TUBITAK) for its PhD scholarship support.

WOS: 000404458100052, PubMed ID: 27901202, This study was designed to determine the in vivo performance of three different materials as scaffolds for dental pulp stem cells (DPSC) undergoing induced odontogenic differentiation. An odontogenic medium modified by the addition of recombinant human bone morphogenetic protein 2 was used in the experimental groups to induce differentiation. Mesenchymal stem cell medium was used in the control groups. DPSC were transplanted onto the backs of mice via three scaffolds: copolymer of L-lactide and DL-lactide (PLDL), copolymer of DL-lactide (PDL) and hydroxyapatite tricalcium phosphate (HA/TCP). The expression levels of dentin sialo-phosphoprotein (DSPP), dentin matrix protein-1 (DMP1), enamelysin/matrix metalloproteinase 20 (MMP20) and phosphate-regulating gene with homologies to endopeptidases on X chromosome (PHEX) were analysed using RT-PCR. The expressions in the experimental groups were compared to those in the control groups. The transcript expressions at 6 and 12 weeks were significantly different for all scaffolds (p < 0.05), except for the expression of DSPP in the PLDL group with regard to the time variable. Although there was a decrease in the expression of enamelysin/MMP20 in PLDL and HA/TCP at 12 weeks, all other expressions increased and reached their highest level at 12 weeks. The highest DSPP expression was in the PDL group (p < 0.05). The highest expression of DMP1 was detected in the HA/TCP group (p < 0.05). The highest expression of PHEX was in the PLDL group (p < 0.05). Consequently, PLDL and PDL seemed to be promising scaffold candidates for odontogenic regeneration at least as HA-TCP, when they were applied with the DPSC induced for odontogenic differentiation., Ege University's Scientific Research FoundationEge University [2010-Dis-006], This study was supported by Ege University's Scientific Research Foundation (2010-Dis-006). The authors sincerely thank Gul KAYRAK for her help in language editing.

WOS: 000368755700005, PubMed ID: 26510940, This study was designed to evaluate the cytotoxicity of four dentin bonding agents and the effects of an antioxidant addition. Group A: G-aerial Bond, Group B: Optibond All in One, Group C: Gluma Self Etch and Group D: Clearfil S-3 Bond were added to the medium using extract method. The cells were cultured with or without resveratrol (RES) addition. MTT, reactive oxygen species (ROS), DCF, Comet and 8-OHdG measurements were performed. The agents had a dose-dependent (1:1>1:10>1:20) cytotoxic effect. Considering 1:10 concentration; Group D at 1 h (p, Ege University's Scientific Research FoundationEge University [2012-Dis-022], This study was supported by Ege University's Scientific Research Foundation (2012-Dis-022). The authors would also like to thank Prof. S. Ismet Deliloglu Gurhan and Mehmet Ozgun Ozen for their kind help in providing cells.

We explored the effects of early and late application of botulinum-A toxin (BTX-A) on reservoir function and histological bladder changes in spinal cord injured rats.The study was done in 30 Sprague-Dawley rats randomly allocated into 5 groups. Group 1 of 6 rats underwent sham operation only. Group 2 of 6 rats underwent spinal cord transection. Group 3 of 6 rats underwent spinal cord transection followed by BTX-A application into the detrusor muscle 7 days later. Group 4 of 6 rats underwent spinal cord transection, followed by BTX-A application into the detrusor muscle 28 days later. Group 5 of 6 rats underwent spinal cord transection followed by saline injection into the detrusor muscle after 28 days. Spinal cord injury was created by transecting the cord at the T9 to T10 level. All rats underwent cystometric examination initially and on day 42 before sacrifice. The bladders were removed and examined histologically for fibrosis and hyperplasia.On cystometric examination BTX-A caused an improvement in baseline pressure, and the frequency and amplitude of uninhibited detrusor contractions (p0.001). No significant differences were observed in maximal bladder capacity or urethral opening pressure (p0.05). Histologically BTX-A led to decreased fibrosis and hyperplasia (p0.001). No significant differences were found between histological or cystometric among the groups with respect to receiving BTX-A in the early and late periods (p0.05).BTX-A has a functional and histological healing effect on detrusor hyperreflexia subsequent to spinal cord injury in rats. Although administering BTX-A in the early period had better quantifiable functional and histological outcomes compared to the late period, the difference was not statistically significant.

Cholinergic and dopaminergic systems are involved in spatial memory and are modulated by nitric oxide (NO); NO has well documented effects on place learning in rodents. The aim of the present study was to investigate the effect of NOS inhibition on place learning in the water maze and to evaluate the relationships between NOS inhibition, learning performance, dopamine (DA) D2 and muscarinic acetylcholine (mACh) receptors. Male Sprague–Dawley rats received the NOS inhibitor Nω-Nitro- l -Arginine ( l -NA), or saline and were trained in the water maze. Rats that were not trained, but received the same treatments were also included. Following treatments with or without water maze training, [ 3 H]-QNB and [ 3 H]-spiperone binding in cortex, striatum and hippocampus were determined to assess the effects of NOS inhibition and/or learning on DA D2 and mACh receptor regulation. The overall results of the present study showed that: (1) NOS inhibition impairs performance in the MWM; (2) NOS inhibition does not affect specific binding to DA D2 (striatum and hippocampus) and mACh (cortex and hippocampus) receptors; (3) MWM training lowers D2 and mACh receptor binding in cortical regions.

Meconium aspiration syndrome (MAS) frequently results in inactivation of surfactant, persistent pulmonary hypertension (PPHN) and respiratory failure among newborn infants. Inflammation and inflammatory mediators play an important role in MAS. Since alveolar macrophages are thought to be very important cells in the pathogenesis of various inflammatory diseases, we evaluated whether meconium could stimulate rat alveolar macrophages to generate platelet-activating factor (PAF) and tumor necrosis factor (TNF)-alpha in vitro. We also examined the response to A23187 (calcium ionophore), 1-0-Hexadecyl-2-acetyl-sn-glycero-3-phosphocholine (synthetic PAF) and dexamethasone on meconium-induced release of PAF and TNF-alpha. PAF and TNF-alpha concentrations from supernatant fluid were measured after high-performance liquid chromatography purification by specific radioimmunoassay, and TNF-alpha concentrations were determined by using an enzyme-linked immunosorbent assay. Our results showed that alveolar macrophages exposed to meconium could enhance PAF and TNF-alpha production in a dose (0.1, 1, 5 and 10%, P

Systemic kainic acid (KA) administration to rats triggers wet dog shakes (WDS) followed by epileptic seizures. Although WDS are often associated with the occurrence of seizures, we have recently shown that following nitric oxide (NO) synthesis inhibition, the number of WDS decreased; subsequently the onset of seizure activity was shortened, and the number of convulsions was increased. Somatostatin (SS), whose release appears to be controlled by NO, inhibits seizure activity. There are sex differences in seizure susceptibility as well as in SS and NO activities in brain. The present study was undertaken to assess the effect of octreotide (OC), a stable SS analogue, on KA-induced WDS and seizures in rats, with emphasis on possible sex differences. WDS and seizures were induced by KA in male and female (proestrus) Sprague Dawley rats; OC or saline was injected 30 min before KA and the behavior was monitored for 120 min after KA. Octreotide increased the number of WDS and decreased the number of convulsions; this effect was more pronounced in males. Onset of KA-induced seizure activity was earlier in females than males; however, there was no effect of OC on seizure latency. Seizure activity started after the termination of WDS. These results show OC has opposite effects on WDS and convulsions, in that it stimulates the former and inhibits the latter. These results support our previous findings that WDS and seizure activity involve separate mechanisms and suggest that WDS may have an inhibitory effect on limbic seizures.

Oxidative stress plays a central role in the pathogenesis of Parkinson's disease. Recently, it has been shown that glycogen synthase kinase 3beta (GSK-3β) is activated in various MPP+/MPTP models of Parkinsonism and some potent GSK-3β inhibitors such as lithium and TDZD-8 protect against MPP+-induced cell death in SH-SY5Y cells. Tideglusib, a thiazolidinedione compound, is a non-ATP competitive inhibitor of GSK-3β and an agonist of peroxisome proliferator-activated receptor gamma (PPARγ). Recently, dual effect of tideglusib is thought to have a role in neuroprotection. The aim of the present study was to evaluate the antioxidant effects of tideglusib in MPP+-induced dopaminergic neuronal loss in SH-SY5Y cells. Reactive Oxygen Species (ROS) production was measured by DCF-DA assay and SOD, catalase and GPx enzyme activities were analyzed following treatments. In the present study, MPP+-induced cell death and ROS generation were significantly attenuated by tideglusib. Although we could not observe any significant alteration in SOD activities following tideglusib treatment against MPP+, both catalase and GPx enzyme activities were significantly increased in tideglusib-treated cells. Our study provides the evidence that both GSK-3β inhibition and PPARγ activation may exert neuroprotection through increasing endogenous cellular antioxidant defenses in cell culture model of Parkinson's disease.

Traumatic spinal cord injury (SCI) can lead to post-traumatic inflammation, oxidative stress, motor neuron apoptosis, necrosis and autophagy of tissue. To promote and enhance recovery after SCI, recent development of devices and therapeutic interventions are needed. The aim of the present study was to investigate the possible role of C-terminal domain of tetanus toxin (Hc-TeTx) on cell death mechanisms including apoptosis and autophagy following SCI. Thirty five adult rats were divided into five groups (n=7 each) as follows: control, sham, trauma (SCI), SCI+Hc-TeTx and SCI+methylprednisolone groups. The functional neurological deficits due to the SCI were assessed by behavioral analysis using the BBB open-field locomotor test. The alterations in pro-/anti-apoptotic and autophagy related-protein levels were measured by western blotting technique. In our study, Hc-TeTx promotes locomotor recovery and motor neuron survival of SCI rats. Hc-TeTx decreased expression of bax, bad, bak, cleaved caspase-3, Ask1 and autophagy-related proteins including Atg5 and LC3II in brain. Our study provides an evidence that cell death mechanisms play critical roles in SCI and non-toxic peptides may exert protective effect and decrease cell death following SCI.

Currently, hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide. However, the studies focusing on molecular etiology of HCC are limited. The Nrf2 signaling pathway can protect cells from a variety of toxicants and carcinogens by increasing the expression of a number of cytoprotective genes.The idea of using G. lucidum for cancer treatment is based on numerous laboratory and preclinical studies with cancer and immune cells as well as animal models demonstrating various biological activities in vitro and in vivo. The aim of our study is to evaluate the potential antioxidant role of G. lucidum in HCC. For this purpose, analysis of Nrf2 levels and cell cycle arrest were done. The most effective concentration of G. lucidum were found at 1/5 dilution. The changes in cytoplasmic/nuclear Nrf2 protein levels following G. lucidum extracts (1:5 or 1:10) treatments for 24, 48 or 72 h were observed. 1/10 and 1/5 diluted of G. lucidum induced G0/G1 cell cycle arrest in HCC. Consistently, the cells distributed in S phase were significantly reduced.In conclusion, our findings suggest that G. lucidum has a potential as an anticancer agent and. our data support the importance of the clarification of the molecular mechanisms of phytochemicals-induced Nrf2 activation.

Background N-methyl- d -aspartate (NMDA) receptors are major pharmacological targets to prevent or reduce the progression of neurodegenerative diseases. Successful therapy with NMDA receptor antagonists in humans has been limited by the severe side effects of complete receptor blockade. The aim of the present study was to investigate the possible protective effects of tideglusib against NMDA receptor overactivation in neural stem cells. Methods We measured the alteration in membrane integrity, free radical generation, intracellular Ca 2+ accumulation, mitochondrial membrane potential (MMP)/mitochondrial morphology and glycogen synthase kinase-3 (α/β isoforms and phospho-GSK-3α/β) protein expression levels following treatments. Results NMDA treatment, with or without d -serine, significantly increased LDH leakage and triggered cell death in neural stem cells. Reactive oxygen species (ROS) formation and intracellular Ca 2+ levels were increased following NMDA receptor overactivation. The significant reduction in MMP was found in NMDA/ d -serine-treated cells. Tideglusib significantly decreased ROS production and membrane degradation, but did not change intracellular Ca 2+ levels following NMDA receptor activation. Both in the presence or in the absence of NMDA/ d -serine, tideglusib increased MMP and the levels of phospho-GSK-3β in NSCs. Moreover, GW9662 (a peroxisome-proliferator-activated receptor gamma (PPARγ) antagonist) treatment significantly inhibited the protective effect of tideglusib in NMDA/ d -serine-treated cells. Conclusion Our study provides the evidence that GSK-3β and PPARγ may be directly involved in pathways leading to NMDA receptor-induced cell death and that the inhibitors including tideglusib may exert neuroprotective effect against these receptor overactivation.

To investigate nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and fos expression in spinal cord dorsal horn neurons following noxious peripheral stimulation.Expression of the immediate-early gene c-fos and nitric oxide containing neurons one hour after unilateral formalin injection to the dorsal hind paw was investigated in rat lumbar spinal cord, using fos immunohistochemistry and NADPH-d histochemical techniques. The experiments were performed in 2004 and 2006 at Ege University Center for Brain Research in Izmir, Turkey.In 10 adult male Sprague-Dawley rats, an increase in fos-immunoreactive neurons was observed ipsilateral, and NADPH-d positive neurons equally ipsi- and contralateral to the formalin injection site. Approximately 20% of fos-immunoreactive neurons were NADPH-d positive ipsilateral to the formalin injection, whereas no double labeling was observed in the contralateral side. Also, a close relation of NADPH-d positive processes with fos-immunoreactive nuclei were also observed.The results of this study support the hypothesis that nitric oxide synthase blocking agents may serve as a possible alternative in treatment of hyperalgesia following inflammation and peripheral nerve injury.

Aims The aim of this study is to determine the role of nitric oxide (NO) in neuropathic pain and the effect of embryonic neural stem cell (ENSC) transplantation on NO content in rat spinal cord neurons following spinal cord injury (SCI). Main methods Ninety adult male Sprague–Dawley rats were divided into 3 groups (n = 30, each): control (laminectomy), SCI (hemisection at T12–T13 segments) and SCI + ENSC. Each group was further divided into sub-groups (n = 5 each) based on the treatment substance (L-NAME, 75 mg/kg/i.p.; l -arginine, 225 mg/kg/i.p.; physiological saline, SF) and duration (2 h for acute and 28 days for chronic groups). Pain was assessed by tail flick and Randall–Selitto tests. Fos immunohistochemistry and NADPH-d histochemistry were performed in segments 2 cm rostral and caudal to SCI. Key findings Tail-flick latency time increased in both acute and chronic L-NAME groups and increased in acute and decreased in chronic l- arginine groups. The number of Fos (+) neurons decreased in acute and chronic L-NAME and decreased in acute l -arginine groups. Following ENSC, Fos (+) neurons did not change in acute L-NAME but decreased in the chronic L-NAME groups, and decreased in both acute and chronic l -arginine groups. NADPH-d (+) neurons decreased in acute L-NAME and increased in l -arginine groups with and without ENSC transplantation. Significance This study confirms the role of NO in neuropathic pain and shows an improvement following ENSC transplantation in the acute phase, observed as a decrease in Fos(+) and NADPH-d (+) neurons in spinal cord segments rostral and caudal to injury.

Introduction Erectile dysfunction is common among patients with spinal cord injury (SCI). Aim This study aims to investigate the recovery of penile erectile functions of the rats with spinal cord injury (SCI) following transplantation of endogenous neuronal precursors cell (neuronal restricted precursors [NRP]/glial restricted precursors [GRP]) into the injured area of spinal cord. Methods Twenty‐two rats were experimented in three groups. Group 1 (N = 6): Sham; Group 2 (N = 10): SCI + NRP/GRP transplanted in day 9 after operation; Group 3 (N = 6): SCI + culture medium transplanted in day 9 after operation. Analysis of penile reflexes and cavernosal nerve stimulation studies were performed in day 28 after transplantation for each group. All rats in three groups were then sacrificed and the injured regions of spinal cords underwent histological investigation. Main Outcome Measures These results show improvements to some extent in locomotor and erectile functions although these improvements are far from full functional recovery. Results Cavernosal nerve stimulation resulted in significantly higher intracavernosal pressure in Group 3 (SCI) although there was no difference between Group 1 (sham) and Group 2 (SCI + NRP/GRP). Number of clusters was similar between groups. Number of erections was higher in Group 3 (SCI) than Groups 1 and 2, and number of cups was higher in Group 2 (SCI + NRP/GRP) than the other two groups. Number of flips was similar in Groups 1 and 2 but lower in Group 3. Number of long flips was highest in Group 1 and lowest in Group 3. The differences between groups were significant. Conclusion. This study emphasized the healing potential of NRP/GRP transplantation following experimental SCI. However, further experimental and clinical studies are required to advance this treatment modality. Temeltas G, Dagci T, Evren V, and Lekili M. Effects of neuronal and glial restricted precursor cells transplantation on erectile function after experimentally induced spinal cord injury.

WOS: 000268125600014, PubMed ID: 18637713, The oxidative mechanisms of injury-induced damage of neurons within the spinal cord are not very well understood. We used a model of T8-T9 spinal cord injury (SCI) in the rat to induce neuronal degeneration. In this spinal cord injury model, unilateral avulsion of the spinal cord causes oxidative stress of neurons. We tested the hypothesis that apurinic/apyrimidinic endonuclease (or redox effector factor-1, APE/Ref-1) regulates this neuronal oxidation mechanism in the spinal cord region caudal to the lesion, and that DNA damage is an early upstream signal. The embryonic neural stem cell therapy significantly decreased DNA-damage levels in both study groups-acutely (followed up to 7 days after SCI), and chronically (followed up to 28 days after SCI) injured animals. Meanwhile, mRNA levels of APE/Ref-1 significantly increased after embryonic neural stem cell therapy in acutely and chronically injured animals when compared to acute and chronic sham groups. Our data has demonstrated that an increase of APE/Ref-1 mRNA levels in the caudal region of spinal cord strongly correlated with DNA damage after traumatic spinal cord injury. We suggest that DNA damage can be observed both in lesional and caudal regions of the acutely and chronically injured groups, but DNA damage is reduced with embryonic neural stem cell therapy.

1. Glial cells are the most abundant cell population in the central nervous system. The aim of this study was to examine the effects of melatonin, 7-nitroindazole, and riluzole on glutamate toxicity in primary glial cell culture. 2. Glutamate toxicity was induced by addition of 100 microM glutamate to the cultures, and then 100 microM melatonin, 500 microM 7-nitroindazole, and 10 (M riluzole were administered in different groups. Lactate Dehydrogenase activity and nitrite levels were determined at the 1st, 6th, and 24th h. 3. Melatonin, 7-nitroindazole, and riluzole decrease Lactate Dehydrogenase activity at the 1st, 6th, and 24th h (at all hours, p0.05). Nitrite levels were decreased by melatonin and riluzole at the 1st, 6th, and 24th h. 4. In this study, we observed that melatonin, 7-nitroindazole, and riluzole are effective as protective agents on glutamate toxicity in mixed glial cells.

Object. Spinal cord injury (SCI) is a complex pathological entity, the treatment of which requires a multipronged approach. One way to integrate different therapeutic strategies for SCI is to develop implantable scaffolds that can deliver therapies in a synergistic manner. Many investigators have developed implantable “bridges,” but an important property of such scaffolds—that is, mechanical compatibility with host tissues—has been neglected. In this study, the authors evaluated the results of implanting a mechanically matched hydrogel-based scaffold to treat SCI. Methods. A nonbiodegradable hydrogel, poly(2-hydroxyethylmethacrylate) (PHEMA), was engineered using thermally initiated free radical solution polymerization. Two groups of 12 adult Sprague—Dawley rats underwent partial cervical hemisection injury followed by implantation of either PHEMA or PHEMA soaked in 1 µg of brain-derived neurotrophic factor (BDNF). Four rats from each group were killed 1, 2, or 4 weeks after induction of the injury. Immunofluorescence staining was performed to determine the presence of scarring, cellular inflammatory responses, gliosis, angiogenesis, and axonal growth in and around the implanted scaffolds. Conclusions. The implanted PHEMA with 85% water content had a compressive modulus of 3 to 4 kPa, which matched the spinal cord. Implanted PHEMA elicited modest cellular inflammatory responses that disappeared by 4 weeks and minimal scarring was noted around the matrix. Considerable angiogenesis was observed in PHEMA, and PHEMA soaked in BDNF promoted axonal penetration into the gel. The authors conclude that mechanically engineered PHEMA is well accepted by host tissues and might be used as a platform for sustained drug delivery to promote axonal growth and functional recovery after SCI.