Lithium (Li) was presented as a protective agent in neuron degeneration which is an important process in neurodegenerative diseases. The aim of this study was to determine the effect of certain amounts of Li on neuroblastoma cells by considering several specific genes which act on neuroprotection. Li solutions were prepared as 1 μM, 15 μM (low dosage), 30 μM and 45 µM (high dosage) concentrations then applied to neuroblastoma cell line. XTT and trypan blue assays were performed to determine the cell proliferation and viability, respectively. mRNA expression levels of NES, BDNF, GRIN2A, LRRK2, PRKN, and SNCA were detected by quantitative real time polymerase chain reaction (qRT-PCR). Cell viability detected as significantly increased in cells treated with low dosage Li however, it was significantly decreased in high dosage applied cells compared to untreated control. In addition, cell proliferation ratios were significantly decreased in high dosage applied cells compared to control. It was demonstrated that mRNA expression levels of several genes (NES, LRRK2, PRKN) were significantly upregulated. Regarding to BDNF, expression of the gene was significantly upregulated in the cells only treated with very low amount of Li. However, no significant data could be obtained for GRIN2A. Furthermore, mRNA expression level of SNCA was determined as significantly downregulated compared to control. Statistically significant expression of NES, LRRK2, PRKN, BDNF and SNCA genes due to the variable Li concentrations applied to cells suggests that Li acts on transcriptional regulation of certain genes associated with neuronal survival. These findings support that dose dependent Li treatment might have a protective effect for neurodegenerative diseases.

Aims:Circadian rhythm of human body is regulated by melatonin which affects through to hypothalamic area of brain.Human clock genes (Clock, Bmal1, etc.) also actively participate in the circadian rhythm.Cell viability techniques reflect the number of living cells in a population.In this manuscript, human neuroblastoma cell viability was found in melatonin usage.The clock gene expression differences on the same panel were analyzed.Methods:In cell viability studies, MTT and trypan blue assays were used with melatonin solutions (in 0.25mM, 0.50mM, 1mM concentrations) on neuroblastoma cells.In the expressions studies, Clock, Bmal1, Per and Cry genes were studied for gene expressions by using melatonin on RT-PCR.Results:The melatonin' LD50 was found as 1.2 mM on human neuroblastoma cells.We found that, 81±1.2%,80±0.9%,73±1.2%,73±1.1% cells were viable in control, 0.25mM,0.50mM, 1mM melatonin concentrations respectively.In high melatonin concentrations (0.5mM and 1mM), Clock and Bmal1 expressions were found higher than control (p<0.05).In low melatonin concentration (0.25mM), no difference in expression was found on Clock and Bmal 1 genes (p≥0.05).No difference in expressions was found in Per and Cry genes by using melatonin (p≥0.05).Conclusions:Melatonin in high concentrations decreased neuroblastoma cell viability.In the same concentrations, melatonin increased the Clock and Bmal1 genes' expressions.Melatonin in low concentration did not chanced cell viability.In the same concentration, no effect on Clock and Bmal1 genes' expressions were observed.We observed no relationship between Per and Cry genes' expressions and melatonin usage.

Objective: Monosodium glutamate appear as a food additive that listed in the group of flavor enhancers is continuously used in various types of foods. Monosodium glutamate is the sodium salt of glutamic acid, one of the most abundant naturally-occurring non-essential amino acids. U.S. Food and Drug Administration reported that monosodium glutamate is safe for use in food. However, the excessive use of monosodium glutamate causes dizziness, nausea and vomiting. Due to reports, it causes obesity. In this study it was aimed to find out the cytotoxicity of monosodium glutamate and the role on the selected genes, which have role on obesity. Method: In our study, mice were used which are known to be the most sensitive in terms of toxicity. The cytotoxic effects of monosodium glutamate on mouse mesenchymal stem cells at monosodium glutamate doses below the blood dose ( 100-130 µmol/dl) that reported as the neuronal damage threshold in mice were studied . Furthermore leptin-lep and ghrelin / obestatin in prepropeptide-GHRL gene expressions in order to find out the role of monosodium glutamate in obesity were analyzed. Results: Monosodium glutamate below the toxic dose does not have a cytotoxic effect on mouse mesenchymal stem cells. Also no expression change in applied monosodium glutamate doses was observed in genes which are known to be associated with obesity. Conclusions: Our results support that monosodium glutamate has no toxic effect on stem cells in uses in certain doses.

SUMMARY Aim/Backround: Monosodium glutamate as a food additive is a substance added to food to preserve flavor or enhance its taste and appearance. Monosodium glutamate is the sodium salt of glutamic acid, one of the most abundant naturally-occurring non-essential amino acids. U.S. Food and Drug Administration reported that monosodium glutamate is safe for use in food. However, the excessive use of monosodium glutamate causes dizziness, nausea and vomiting. Due to reports, it causes obesity. We aimed to find out the cytotoxiciy of monosodium glutamate and the role on the selected genes, which have role on obesity. Methods: In our study, we used mice which are known to be the most sensitive in terms of toxicity. We were studied the cytotoxic effects of monosodium glutamate on mouse mesenchymal stem cells at monosodium glutamate doses below. Also we analyzed leptin-lep and ghrelin / obestatin in prepropeptide-GHRL gene expressions in order to find out the role of monosodium glutamate in obesity. Result: Monosodium glutamate below the toxic dose does not have a cytotoxic effect on mouse mesenchymal stem cells. Also no expression change observed in genes which are known to be associated with obesity. Conclusion: Our results support that monosodium glutamate has no toxic effect on stem cells in uses in certain doses. Key words: Monosodium glutamate, mouse mesenchymal stem cells, leptin gene, ghrelin/obestatin prepropeptide gene. Running Title: MSG has no ctotoxicity

Amac: Usnik asit likenlerde bulunan halen rolu tam olarak ortaya konamamis bir sekonder metabolittir. Usnik asit iceren liken ekstraktlari tipta, parfum yapiminda, kozmetikte ve ekolojide kullanilmaktadir. Usnik asit insan akciger kanserinde hucre buyumesini durdurmakta, hucre dongusunun durdurulmasi ve apopitozu aktive etmektedir. Calismamizda ayni hucre hattinda usnik asitin antitumoral etkisi bakilmistir. Yontem: Hucre apopitozunda rolu olan APOPT1, CYCS, APAF1, CASP3/9, TNF , BCL2 ve BCL2L1 genlerinin ekspresyon analizine bakildi. Bulgular: Usnik asitin APOPT1, CYCS, APAF1, CASP3, CASP9 genleri uzerinde uyarici etkisi oldugu saptandi. Sonuc: Veriler usnik asitin kanser hucreleri uzerinde mitokondrial apopitotik genler araciligi ile antitumoral etki yaptigini ortaya koymaktadir.

ÖZET Sorefenib ısı şok protein70 genini multiple myelom (RPMI-8226) ve plazma hücreli lösemi (ARH-77) hücre hatlarında inhibe eder Sorafenib solid yapıdaki tümörlerde kullanılan antikanser ilaçtır. Bu ilaç hücrede pek çok tirozin kinazı inhibe eder ve hücreyi apopitoza yönlendirir. Multipl myeloma ve plazma hücreli lösemi hastalık olarak plazma hücreli diskraziler içerisindedir. Halen her iki hastalığın tedavisinde de zorluklar vardır. Isı şok proteinler hücre için önemli olup protein katlanmasında, hücrelerin stersen korunmasında rolü alırlar. Isı şok proteinler birçok kanser hücrelerinde artmış olarak bulunur. Bu proteinlerin aşırı yapımı hücre büyümesi ve metastazında önemlidir. Çalışmamızda sorafenibin sitotoksik etkisi multipl myelom ve plazma hücreli lösemi hücre hatlarında tripan blue ve MTT yöntemleri ile bakılmıştır. MTT yöntemi ile RPMI-8226 hücrelerinde LD50 dozu 12 μM, ARH-77 hücrelerinde 9 μM bulunmuştur. Tripan blue ile yapılan hücre canlılık oranları analiz sonuçları MTT sonuçlarını desteklemektedir. Multipl myelom hücre hattında %89 oranında, plazma hücreli lösemi hücre hattında %80 oranında canlı hücre saptanmıştır. Çalışmamızda ısı şok protein70 ve 90 gen ekspresyon sonuçları da aynı zamanda bu hücre hatlarında çalışılmıştır. Sorafenibin ısı şok protein70 gen ekspresyonunu her iki hücre hattında da inhibe ettiği saptanmıştır. Bu sonuç, sorafenibin sitotoksik etkisinin apopitozu multipl myelom ve plazma hücreli lösemi hücre hatlarında ısı şok protein70 geni üzerinden etkileyerek oluşturduğunu göstermektedir.

Amac: Monosodyum glutamat lezzet arttiricilar grubunda gida katki maddesi olarak cesitli yiyeceklerde kullanilmaktadir. Monosodyum glutamat glutamik asidin sodyum tuzudur ve dogal yollarla en fazla olusan esansiyel olmayan aminoasitlerden biridir.  Amerikan Gida Ve Ilac Teskilati ( U.S. Food and Drug Administration) monosodyum glutamatin gidada kullaniminin guvenli oldugunu bildirmistir. Ancak asiri kullaniminda bas donmesi, bulanti kusma yapmaktadir. Makalelerde obeziteye neden oldugu bildirilmektedir.  Bu calismada, monosodyum glutamatin sitotoksik ve obezitede rolu olan secilmis bazi genlere olan etkilerinin bulunmasi amaclanmistir. Yontem: Calismamiza toksisite yonunden en hassas organizma oldugu bilinen fareler alinmistir. Monosodyum glutamatin fare mezenkimal hucreler uzerine olan sitotoksik etkisi farelerde noronal hasar olusturdugu bildirilen kan dozunun (100-130 µmol/dl) altindaki dozlar kullanilarak calisilmistir. Ayrica monosodyum glutamatin obezite uzerine olan etkisini bulmak icin “ leptin-lep” ve “ Ghrelin/Obestatin Prepropeptide-GHRL” genlerine ait ekspresyon degisikliklerine bakilmistir. Bulgular: Monosodyum glutamatin toksik dozun altinda fare mezenkimal kok hucre uzerinde bir sitotoksik etkisi olmadigi bulunmustur. Uygulanan monosodyum glulatamat dozlarinda hucrede obezite ile ilgili oldugu bilinen genlerin ekspresyonlarini degistirmedigi saptanmistir. Sonuc : Sonuclarimiz monosodyum glutamatin gidada belli dozlarda kullaniminin kok hucreler uzerine toksik bir etkisi olmadigini destekler niteliktedir.

SUMMARY Usnic acid is a secondary metabolite in lichens whose role has not been completely elucidated. Lichen extracts containing usnic acid have been utilized in medicine, perfumery, cosmetics, and ecology. In a recent manuscript, usnic acid was reported as an inhibitor in cell growth, inducing the cell cycle arrest and apoptosis in human lung carcinoma A549 cells. Herein we analyzed the antitumoral effect of usnic acid on the same cell line. Similar results were observed as in literature. We analyzed the gene expression results in the same panel which have possible role on cell apopitosis. As a result, usnic acid has stimulatory effect on APOPT1, CYCS, APAF1, CASP3, CASP9 gene expressions in our experiments. So, usnic acid has antitumoral effect on cancer cells, affecting on mithocondrial apopitotic genes. Key words: Usnic acid, cell growth, apopitosis, gene, antitumoral affect. OZET Usnik asit likenlerde bulunan halen rolu tam olarak ortaya konamamis bir sekonder metabolittir. Usnik asit iceren liken ekstraktlari tipta, parfum yapiminda, kozmetikte ve ekolojide kullanilmaktadir. Son bir yazida, insan akciger kanser A549 hucre hattinda usnik asit hucre dongusunun durdurulmasi ve apopitozis yoluyla hucre buyumesini durduran bir etken olarak tanimlanmistir. Calismamizda ayni hucre hattinda usnik asitin antitumoral etkisi bakilmistir. Literaturde olan ile benzer sonuclar ortaya konmustur. Ayni panel uzerinde hucre apopitozunda rolu olan genlerin ekspresyon analizine bakildi. Sonuc olarak, usnik asitin APOPT1, CYCS, APAF1, CASP3, CASP9 genleri uzerinde uyarici etkisi oldugu saptandi. Veriler usnik asitin kanser hucreleri uzerinde mitokondrial apopitotik genler araciligi ile antitumoral etki yaptigini ortaya koymaktadir. Anahtar Kelimeler: Usnik asit, hucre buyumesi, apopitoz, gene, antitumoral etki. Correspondance To: Prof. Dr. Sefik Guran Saglik Bilimleri Universitesi, Gulhane Tip Fakultesi, Tibbi Biyoloji AD. 06018 Etlik/Kecioren-ANKARA Phone: 03123043551

SUMMARY Usnic acid is a secondary metabolite in lichens whose role has not been completely elucidated. Lichen extracts containing usnic acid have been utilized in medicine, perfumery, cosmetics, and ecology. In a recent manuscript, usnic acid was reported as an inhibitor in cell growth, inducing the cell cycle arrest and apoptosis in human lung carcinoma A549 cells. Herein we analyzed the antitumoral effect of usnic acid on the same cell line. Similar results were observed as in literature. We analyzed the gene expression results in the same panel which have possible role on cell apopitosis. As a result, usnic acid has stimulatory effect on APOPT1, CYCS, APAF1, CASP3, CASP9 gene expressions in our experiments. So, usnic acid has antitumoral effect on cancer cells, affecting on mithocondrial apopitotic genes. Key words: Usnic acid, cell growth, apopitosis, gene, antitumoral affect. OZET Usnik asit likenlerde bulunan halen rolu tam olarak ortaya konamamis bir sekonder metabolittir. Usnik asit iceren liken ekstraktlari tipta, parfum yapiminda, kozmetikte ve ekolojide kullanilmaktadir. Son bir yazida, insan akciger kanser A549 hucre hattinda usnik asit hucre dongusunun durdurulmasi ve apopitozis yoluyla hucre buyumesini durduran bir etken olarak tanimlanmistir. Calismamizda ayni hucre hattinda usnik asitin antitumoral etkisi bakilmistir. Literaturde olan ile benzer sonuclar ortaya konmustur. Ayni panel uzerinde hucre apopitozunda rolu olan genlerin ekspresyon analizine bakildi. Sonuc olarak, usnik asitin APOPT1, CYCS, APAF1, CASP3, CASP9 genleri uzerinde uyarici etkisi oldugu saptandi. Veriler usnik asitin kanser hucreleri uzerinde mitokondrial apopitotik genler araciligi ile antitumoral etki yaptigini ortaya koymaktadir. Anahtar Kelimeler: Usnik asit, hucre buyumesi, apopitoz, gene, antitumoral etki. Correspondance To: Prof. Dr. Şefik Guran Saglik Bilimleri Universitesi, Gulhane Tip Fakultesi, Tibbi Biyoloji AD. 06018 Etlik/Kecioren-ANKARA Phone: 03123043551

Objective:The dental pulp is the part in the center of a tooth made up of living connective tissue and cells called odontoblasts.The vitality of the dentin structure, both during health and after injury, depends on pulp cell activity and the signaling processes that regulate the cell's behavior.Dental pulp tissue has condensed stem cell activity.Dental pulp stem cells are multipotent stem cells that have the potential to differentiate into a variety of cell types.Several publications hav e stressed the importance of the expression of pluripotentiality associated markers: the transcription factors Nanog, Sox2, Oct3/4, SSEA4, CD13, Stro1 are indispensable for the stem cells to divide indefinitely without affecting their differentiation poten tial (self renewal) capacity.Progesterone is a steroid hormone leading to menstrual cycle and gestation.There is a widespread rumor among people that pregnancy causes toothy loss.Method: So, progesterone was applied in different concentrations on human dental pulp cells in cell culture.Cell viability assay was applied 24 th hour later with trypan blue.RNA isolation, cDNA synthesis and Real Time PCR analysis were applied on selected transcription factors (Nanog and Oct4 (POU5F1) genes) which have role on steamness of stem cells.Gene expression analyses results were correlated with the cell viability assay results.Results: Cell viability assay results were 80% viable in control, 82% viable in 7 ml progesterone application, 81% viable in 14 ml progesterone application, 83% viable in 21 ml progesterone application.Due to our findings, progesterone in different concentrations did not chance the cell viability in dental pulpa cells .On gene expression analyses, preliminary results supported that high concentrations of progesterone enhance the gene expressions of steamness genes (Nanog, and Oct4) in dental pulp cells.Conclusions: So, progesterone did not change cell viability in high concentrations.We assume that progesterone did not affect apopitotic pathways on dental pulp cells.On the other hand, high gene expressions of Nanog, and Oct4 in high doses of progesterone represent the importance of progesterone hormone in dental pulp cells including stem cells maintenance.

Objective: Centaury oil (the extract of Hypericum perforatum obtained with olive oil) is traditionally used for the treatment of wounds in Turkey.Centaury oil has analgesic and antiinfectious properties beside wound healing effect.There are many studies about the positive effect of this extract on wound healing in literature.But there is no mention about the animal studies that show which pathways are involved in.Method: In our study the effect of centaury oil in early stages (first 7 days) and late stages (second 7 days) of wound healing was showed on the mice (mus.musculus) by creating sterile sutures on their back.In addition, the gene expression profiles of VEGFA, VEGFB, VEGFC, PDGFB and FGF2 genes which have roles in wound healing via angiogenesis were studied by RT-PCR method.Results: Due to our results, centaury oil has positive effects on the surface of the wounds in early stages but has no effect on late stages.The gene expression levels of VEGFA, VEGFB, VEGFC and FGF2 were increased in early stages of wound healing but only VEGFA gene expression level was increased in late stages.The gene expression findings were correlated with morphological findings in both early and late stages of wound healing.Conclusions: According to our morphological results, centaury oil can be useful if it is used in the first week of the wound.At the end of our study, our findings are important for exploring the effect of centaury oil on angiogenesis via VEGFA, VEGFB, VEGFC and FGF2 genes in early stages of wound healing.

Centaury oil (the extract of Hypericum perforatum obtained with olive oil) is traditionally used for the treatment of wounds in Turkey. Centaury oil has analgesic and anti-infectious properties beside wound healing effect. There are many studies about the positive effect of this extract on wound healing in literature. But there is no mention about the animal studies that show which pathways are involved in. In our study the effect of centaury oil in early stages (first 7 days) and late stages (second 7 days) of wound healing was showed on the mice (mus. musculus) by creating sterile sutures on their back. Due to our results, centaury oil has positive effects on the surface of the wounds in early stages but has no effect on late stages. This information is explained as centaury oil can be useful if it is used in the first week of the wound. In addition, the gene expression profiles of VEGFA, VEGFB, VEGFC, PDGFB and FGF2 genes which have roles in wound healing via angiogenesis, were studied by RT-PCR method. As a result, the gene expression levels of VEGFA, VEGFB, VEGFC and FGF2 were increased in early stages of wound healing but only VEGFA gene expression level was increased in late stages. The gene expression findings were correlated with morphological findings in both early and late stages of wound healing. Our findings are important for exploring the effect of centaury oil on angiogenesis via VEGFA, VEGFB, VEGFC and FGF2 genes in early stages of wound healing.

SUMMARY Objective : Embryonal brain development including neurogenesis and gliogenesis requires the concerted actions of numerous genes that are regulated at multiple levels. As known, transcription factors have roles in neural and glial development. But the roles of these factors in brain development in different gestational periods are not clear. Method : The gene expression profiles of NGN 3, Klf 4, c-Myc, Nanog, Oct ¾, GATA 4, Sox 2 genes had been searched in brain tissues of mice fetus at the end of the first and the third trimesters. Results : The results were correlated with the results obtained from brain tissue of adult mouse. At the end of the first trimester, higher gene expression levels were found due to internal control in NGN 3, Klf 4, c-Myc, Nanog, Oct ¾. Similar gene expression levels were observed in GATA 4. Dramatically lower gene expressions were analyzed in Sox 2. In each gene, the highest gene expression levels were found in the first trimester. The expression level of each gene was decreased due to time period during the pregnancy. Interestingly, the gene expression levels of NGN 3 and Klf 4 were extremely high due to control gene level in the first and third trimesters in our series. Conclusion: According to these results, NGN 3 and Klf 4 genes have important roles in fetal mice brain development during the gestation period. Keywords: Brain development, neurogenesis, gliogenesis, gene expression, NGN 3, Klf 4 OZET Amac : Norogenez ve gliogenezden olusan embriyonal beyin gelisimi farkli asamalarda cesitli genlerin katimli ile duzenlenen bir surectir. Noronal ve glial doku gelisiminde bazi transkripsiyonal faktorlerin rolu oldugu bilinmektedir. Ancak bu transkripsiyonal faktorlerin beyin gelisiminde hamileligin farkli donemlerindeki rolleri tam olarak acik degildir. Yontem: Calismamizda fare fetal beyin dokularinda NGN 3, Klf 4, c-Myc, Nanog, Oct ¾, GATA 4, Sox 2 transkripsiyonal faktor genlerinin ekspresyon duzeylerine hamileligin birinci ve ucuncu trimester sonlarina da donemlerde bakilmistir. Bulgular: Elde edilen sonuclar eriskin fare beyninden elde edilen sonuclarla karsilastirilmistir. Birinci trimester sonunda NGN 3, Klf 4, cMyc, Nanog, Oct ¾ genlerinde kontrole gore yuksek gen ekspresyon duzeyleri saptanirken, GATA 4 geninde kontrole gore benzer gen ekspresyon duzeyi bulunmustur. Sox 2 geninde ise cok dusuk gen ekspresyon duzeyi tespit edilmistir. Her gene icin en fazla ekspresyon duzeyi ilk trimesterda bulunmustur. Bu genlere ait ekspresyon duzeyleri hamilelik suresince zamana bagli olarak azalmistir. Ilginc olarak bizim seride NGN 3 ve Klf 4 gen ekspresyon duzeyleri kontrol gen duzeyine gore birinci ve ucuncu trimetserde oldukca yuksektir. Sonuc : NGN 3 ve Klf 4 genlerinin hamilelik suresince fetal fare beyninin gelisimindeki onemli role sahiptir. Anahtar sozcukler: Beyin gelisimi, norogenez, gliogenez, gen ekspresyonu, NGN 3, Klf 4

Pluripotent stem cells (PSCs) have the capacity to differentiate into any cell type of the body. Therefore, induced pluripotent stem cells (iPSCs) are seen as a promising solution for patient-specific cell therapies. However, the safety is major issue for in vitro methods that are used in induction of pluripotency and also in differentiation of PSCs toward specific cell types. In pioneer studies of iPSC generation, the role of c-Myc has been highlighted as a possible master regulator of pluripotency, but direct c-Myc overexpression is known to prompt drawbacks, especially in human cells. In recent studies, the role of non-protein coding RNA molecules such as microRNAs (miRNAs) has been shown in enhanced reprogramming efficiency. In addition, new reprogramming methods have been ultimately improved by adding miRNAs, in the absence of previous factors. Cross interaction between miRNAs and c-Myc has been also found in differentiation of iPSCs, as well as in reprogramming and self-renewing the pluripotent state. Thence, miRNAs are promising solution for efficiency and safety of iPSC derivation and differentiation methods. The purpose of the present review is to evaluate interaction mechanisms of miRNAs with c-Myc and in iPSC technology.

Objective: Genes are regulated at multiple levels by using nutritional factors during neurogenesis and gliogenesis in brain development. Folic acid (FA)and zinc regulate the expressions of some genes which participate in brain development as nutritional factors. So, we aimed to find the effect of FA and zinc on the expression levels of neurogenin 3 (NGN3), Kruppel-like factor 4 (Klf4), c-Myc, nanog, POU class 5 homeobox 1-Pou5F1 (Oct4), and Nestin in mouse fetal brain tissue in neural plate phase at gestational day 7 (GD7)and in fetal brain tissue at GD20. Methods: All the tissues were cultured primarily. FA and zinc solutions were added. Real time-polymerase chain reaction was performed in RNA obtained from in each sample. Results: FA has no effect on GD7 mouse neural plate tissue and GD20 mouse fetal brain tissue. Zinc has no effect on GD7 mouse neural plate tissue. Zinc increased the expressions of NGN3, Klf4, nanog, Nestin, and Oct4 genes on GD20 mouse fetal brain tissue. Conclusion: This in vitro study represents that zinc is important in the expressions of NGN3, Klf4, Nanog, Nestin, and Oct4 genes in the late phase of pregnancy. The stimulator effect of Zinc on the expression levels of these genes may show us the possible role of zinc in fetal brain development in the late phase of pregnancy.