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    Alcoholic extract of Tarantula cubensis induces apoptosis in MCF-7 cell line.
    (Allied Acad, 2017) Er, Ayse; Corum, Orhan; Corum, Duygu; Hitit, Mustafa; Donmez, Huseyin; Guzeloglu, Aydin
    Tarantula cubensis Alcoholic Extract (TCAE) is a homeopathic agent used for treating many disorders. This study aimed to define the effects of TCAE on the breast carcinoma cell line (MCF-7). After various concentrations (10, 20, 40, 80 and 160 mu l/ml) of TCAE were applied to MCF-7 cells and the human embryonic kidney cell line (HEK293), the cells were incubated for 1, 3, 6, 9, 12, 24 and 48 h, followed by analysis by MTT assays. According to the results of the MTT assays, cells treated with 20 or 40 mu l/ml TCAE for 6 h were applied to apoptosis analysis by flow cytometry. Secreted levels of tumor necrosis factor alpha (TNF alpha), interleukin (IL)-1 beta, IL-6, IL-10, Interferon-gamma (IFN gamma), Transforming Growth Factor beta (TGF beta), and Nuclear Factor-kappa B (NF-kappa B) were measured using ELISAs. TNF alpha and TGF beta levels increased while IL-6 and IL-10 levels fluctuated in MCF-7 cells. In conclusion, our study suggests that TCAE may change the normal cancer physiology and lead to cell death by activating apoptosis in MCF-7 cells.
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    Early pregnancy-related changes in toll-like receptors expression in ovine trophoblasts and peripheral blood leukocytes
    (Elsevier Science Inc, 2017) Kaya, Mehmet Salih; Kose, Mehmet; Guzeloglu, Aydin; Kiyma, Zekeriya; Atli, Mehmet Osman
    In the present study, we aimed to 1) demonstrate the presence of all 10 toll-like receptors (TLRs) in ovine trophoblasts, and 2) investigate the expression profiles of TLR1 - 10 mRNAs in peripheral blood leukocytes (PBLs) in ewes during early pregnancy. For those purposes, ovine trophoblasts (n = 6) were collected from pregnant ewes on day 13. PBLs were collected from non-pregnant (n = 6) and pregnant ewes (n = 17) on days of mating (d) 0 and 18. TLR mRNAs in ovine trophoblasts were visualized by free-floating in situ hybridization (ISH). To assess the expression profiles of TLR1-10 in PBLs, total RNA was isolated and transcribed to cDNA. TLR1-10 mRNA levels were determined by real-time PCR in triplicate. The Relative Expression Software Tool (REST 2009) was used for statistical analysis. We detected mRNAs for TLR2, TLR4, TLR5, TLR6, TLR7, TLR8, and TLR10 but not for TLR1, TLR3, and TLR9 in trophoblasts. TLR2, TLR5, TLR6, TLR7, TLR8, and TLR10 mRNAs were expressed by all trophoblasts, whereas TLR4 mRNA and protein in trophoblasts were more limited. In PBLs, TLR expression did not differ between day 0 and day 18 in non-pregnant ewes; however, ewes in early pregnancy exhibited significantly upregulated expression of TLR2 (23-fold), TLR4 (3.1-fold), TLR6 (1.7-fold), and TLR8 (2.2-fold) on day 18 compared with day 0. In contrast, TLR10 was downregulated (2-fold) on day 18 by pregnancy. Similar results were also obtained for TLR2, TLR4, TLR6, TLR8 and TLR10 from the comparison between day 18 non pregnant and day 18 pregnant groups. According to these results, the presence of TLRs in early ovine trophoblasts suggests that these cells play an immunological role at the maternal fetal interface. The results also suggest that tight regulation of some components of TLRs in PBLs due to embryo- and/or pregnancy related factors is necessary for successful establishment of early pregnancy in ewes. (C) 2017 Elsevier Inc. All rights reserved.
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    Evaluation of genes involved in prostaglandin action in equine endometrium during estrous cycle and early pregnancy
    (Elsevier, 2010) Atli, Mehmet O.; Kurar, Ercan; Kayis, Seyit A.; Aslan, Selim; Semacan, Ahmet; Celik, Sefa; Guzeloglu, Aydin
    The aim was to evaluate expression of genes involved in the biosynthesis of prostaglandins (PTG), Prostaglandin H Synthase-1 (PTGS1) and PTGS2, PGF synthase (PTGFS), and PGE synthase (PTGES), PGF receptor (PTGFR). PGE receptors (PTGER2 and FTGER4), prostaglandin transporter (SLCO2A1) and hydroxyprostaglandin dehydrogenase-15 (HPGD). Endometrial biopsies were obtained from mares on day of ovulation (d0, n = 4), late diestrus (LD, n = 4), early luteolysis (EL n = 4) and after luteolysis (AL n = 4) during the cycle. Stages of the cycle were confirmed by plasma progesterone concentrations measured daily and ultrasound examinations. Biopsies were also taken on days 14 (P14; n = 4), 15 (P15. n = 4), 18 (P18, n = 4) and 22 (P22; n = 4) of pregnancy. Relative mRNA expressions were quantified using real-time RT-PCR. A mixed model was fitted on the normalized data and least significant difference test (alpha = 0.05) was employed. Expression of PTGS1 mRNA was low throughout the estrous cycle and early days of pregnancy, but upregulated on P18 and P22. PTGS2 expression was increased on EL but it was suppressed by pregnancy on P15, P18, and P22. PTGFS expression was upregulated in both cyclic and pregnant mares compared to d0 and its level was the highest on LD. PTGFR expression was transiently increased on LD and EL and was suppressed during early pregnancy. Both PTGES and PTGER2 expressions were increased on LD, El, and early pregnancy, but were decreased after the luteolysis in cyclic mares as they remained high on P18 and P22. PTGER4 expression did not change throughout the cycle and early pregnancy. Levels of HPGD and SLCO2A1 were significantly increased only on P22. In conclusion, PTGS2 expression increases around the time of luteolysis and concurrent upregulation of PTGFS and PTGES indicates that equine endometrium has increased capability of PTG production around the time of luteolysis. However, pregnancy reduces PTGS2 expression, but maintains the high levels of PTGES during early pregnancy along with PTGER2 while PTGFR expression was suppressed. These findings suggest that possible luteotrophic action of PGE(2) is required in early equine pregnancy. PTGS1 is only upregulated later in the early pregnancy suggesting that it is not involved in luteolysis, but could be the main PTGS enzyme at this time during early pregnancy. An increase in HPGD and SLCO2A1 levels on P22 indicates a tight regulation of PTG action by pregnancy. (C) 2010 Elsevier B.V. All rights reserved.
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    Expression of enzymes and receptors of leukotriene pathway genes in equine endometrium during the estrous cycle and early pregnancy
    (Elsevier Science Inc, 2013) Guzeloglu, Aydin; Atli, Mehmet O.; Kurar, Ercan; Kayis, Seyit A.; Handler, Johannes; Semacan, Ahmet; Aslan, Selim
    The aims of the present study were to elucidate the expression profiles of leukotriene (LT) pathway mRNA transcription and to determine the possible interaction of LT and prostaglandin (PTG) pathways genes in equine endometrium during the estrous cycle and early pregnancy. Endometrial biopsies were obtained from mares on the day of ovulation (d0), at late diestrous (LD, n = 4), and after luteolysis in the estrus phase (AL n = 4) of the cycle. Biopsies were also taken on Days 14 (P14; n = 4), 18 (P18, n = 4), and 22 (P22, n = 4) during early pregnancy that were comparable days to cyclic sampling days. A mixed model was fitted on the normalized relative mRNA levels, quantified by qPCR in duplicate, and least significant difference test was employed to detect significantly different group(s). In addition, to determine the degree of contribution of each gene to separation of treatment groups, the multivariate projection method partial least square regression discriminant analysis was used. The expression of 5-lipoxygenase mRNA was greater on d0 and LD, declined at AL, and was suppressed by early pregnancy. Leukotriene A(4) hydrolase mRNA expression increased at LD and during early pregnancy, but Was significantly greater at LD compared with P14. The expression of LT C-4 synthase mRNA was only induced at LD. Cysteinyl leukotriene receptors (CysLT1 and CysLT2) mRNA expressions were decreased by both cyclic changes and early pregnancy, whereas 5-lipoxygenase-activating protein and B leukotriene receptor mRNA expressions were not affected by early pregnancy or stages of the estrous cycle. Partial least square discriminant analysis suggests that LT and PTG pathway enzymes and receptors appear to behave similarly in terms of mRNA expression. In conclusion, the expression profiles of LT pathway genes are demonstrated in equine endometrium for the first time by the present study, and the present data suggest that LT pathway mRNA transcriptions are tightly regulated during early pregnancy in mares. (C) 2013 Elsevier Inc. All rights reserved.
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    Expression of Epidermal Growth Factor (EGF) and Heparin-Binding EGF (HB-EGF) mRNA in Mare Endometrium
    (Kafkas Univ, Veteriner Fakultesi Dergisi, 2012) Atli, Mehmet Osman; Guzeloglu, Aydin; Kurar, Ercan; Kayis, Seyit Ali; Aslan, Selim; Semacan, Ahmet; Celik, Sefa
    EGF and HB-EGF play crucial roles in embryonic development and peri-implantation. In this study, aim was to characterize expression profiles of EGF and HB-EGF in mare endometrium during estrous cycle and early pregnancy. Endometrium biopsies were obtained from mares on day of ovulation (d0), late diestrus (LD) and after luteolysis (AL) in the estrus phase. In pregnant groups, biopsies were taken on days 14 (P14), 18 (P18), 22 (P22) and 60 (P60). Relative expression levels of genes were quantified using real-time RT-PCR. A mixed model was fitted on the normalized data and Least Significant Difference (LSD) test was employed to determine significantly different group(s). EGF mRNA expression was up-regulated at LD compared to d0 while HB-EGF expression was not changed throughout the cycle. EGF expression was also increased during early pregnancy with the highest expression level observed on P60. Similarly, HB-EGF mRNA level was increased on P60. Pregnancy induced EGF expression on P14 and P18 compared to LD and AL whereas expression of HB-EGF was only significantly higher on P18 than that of AL. These results indicate that EGF expression is up-regulated during the cycle at late diestrus when P4 is high and is increased by pregnancy. HB-EGF expression is induced later in the pregnancy. In conclusion, EGF and HB-EGF appear to involve in the events that happen in the mare endometrium during peri-implantation period.
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    Expression of platelet activating factors and its receptor in cat uterus during early pregnancy
    (Ankara Univ Press, 2013) Agaoglu, Ali Reha; Agaoglu, Ozgecan Korkmaz; Kurar, Ercan; Guzeloglu, Aydin; Kayis, Seyit Ali; Atli, Mehmet Osman; Schafer-Somi, Sabine
    Platelet activating factors (PAF1 beta, PAF1 gamma, PAF:ah, PAF-R) have a significant role during mammalian pregnancy process. This study aimed to investigate expression profiles of PAF gene family in cat uterus during pregnancy. Tissue samples were collected via ovariohysterectomy at days 7 (Gr1; n=3), 18-20 (Gr2; n=7), 24-25 (Gr3; n=5) after mating from the different regions of the uterus. Relative expression levels of genes were quantified using real-time qRT-PCR. A mixed model was fitted on the normalized data and Least Significant Difference (LSD) test was employed to determine significant differences between groups. Expressions of PAP genes were assessed in the cat uterus during pregnancy around implantation period; however, no significant differences were detected between the preimplantation and the implantation stage. Similarly, no significant difference concerning mRNA expression was assessed between different parts of the uterus during pregnancy In conclusion, PAP genes are expressed in the cat uterus during early pregnancy. However, this expression is not associated with changes in the uterus during early pregnancy.
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    Expression of Toll-like Receptors (TLRs) Is Regulated by Early Pregnancy in the Mare Endometrium.
    (Soc Study Reproduction, 2010) Atli, Mehmet Osman; Kurar, Ercan; Kayis, Seyit A.; Aslan, Selim; Semacan, Ahmet; Celik, Sefa; Guzeloglu, Aydin
    [Abstract Not Available]
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    Expression of wingless type (WNT) genes and their antagonists at mRNA levels in equine endometrium during the estrous cycle and early pregnancy
    (Elsevier Science Bv, 2011) Atli, Mehmet Osman; Guzeloglu, Aydin; Dinc, Dursun Ali
    WNT signaling pathway plays important roles in reproductive events. Aims were to (1) determine presence of WNT genes and their antagonists in equine endometrium; and (2) to evaluate their expression profiles during early pregnancy. Endometrial biopsies were obtained from mares on day of ovulation (d0, n = 4) and on days of 14 (P14, n = 4), 18 (P18, n = 4), 22 (P22, n = 4) of early pregnancy. Biopsies were also collected from cyclic mares during late diestrus (LD, on day of 13.5-14, n = 4) and after luteolysis in estrus phase (AL on day of 17.5-18, is = 4) of the cycle. PCR was used to detect expression of genes studied and then relative expression levels were quantified using real-time PCR analysis. A mixed model was fitted on the normalized data and least significant difference test (alpha = 0.05) was employed. Eleven WNT genes (WNT2, WNT2B, WNT4, WNT5A, WNT5B, WNT7A, WNT8A, WNT9B, WNT10B, WNT11 and WNT16) and their antagonists (SFRP1, SFRP2, SFRP5, DKK1, DKK2 and WIF-1) were detected in equine endometrium. Compared to d0, WNT2, WNT5B, WNT7A and SFRP1 expressions were downregulated by the pregnancy while DKK1 was upregulated. WNT5A, WNT11 and WIF-1 were upregulated on P14 and P18. but WNT2B increased only on P14. When LD and P14 were compared, level of WNT8A decreased on P14 while increase in WNT4 level on P14 was slightly significant (P < 0.06). Levels of WNT7A and SFRP1 decreased while DKK1 and WIF-1 increased by the pregnancy on P18 compared to AL Moreover, WNT2B, WNT5A, WNT9B, WNT10B, WNT11, WNT16 DKK1 and WIF-1 were upregulated on LD compared to AL whereas WNT4, WNT7A, SFRP1 were downregulated. In conclusion, the results demonstrate that WNT genes and their antagonists appear to be regulated during early pregnancy in equine endometrium possibly due to embryonic factors and/or maternal progesterone. (C) 2011 Elsevier B.V. All rights reserved.
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    Expressions of Insulin-Like Growth Factor-Binding Proteins (IGFBP) in Mare Endometrium during Estrous Cycle and Early Pregnancy.
    (Soc Study Reproduction, 2010) Kurar, Ercan; Atli, Mehmet O.; Kayis, Seyit A.; Arslan, Selim; Semacan, Ahmet; Celik, Sefa; Guzeloglu, Aydin
    [Abstract Not Available]
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    PLS-DA approach for simultaneous evaluation of genes involved in luteolysis in equine endometrium during estrous cycle and early pregnancy
    (Elsevier Sci Ltd, 2011) Guzeloglu, Aydin; Kayis, Seyit A.; Atli, Mehmet O.; Kurar, Ercan; Semacan, Ahmet; Aslan, Selim
    [Abstract Not Available]
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    Rating of putative housekeeping genes for quantitative gene expression analysis in cyclic and early pregnant equine endometrium
    (Elsevier Science Bv, 2011) Kayis, Seyit A.; Atli, Mehmet O.; Kurar, Ercan; Bozkaya, Faruk; Semacan, Ahmet; Aslan, Selim; Guzeloglu, Aydin
    The aim was an evaluation of a set of housekeeping genes (HKGs) to be used in the normalization of gene expression in the equine endometrium. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine ribosyl transferase 1 (HPRT1), ubiquitin B (UBB), tubulin alpha 1 (TUBA1), ribosomal protein L32 (RPL32), beta-2-microglobulin (B2M), 18S rRNA (18S), and 285 rRNA (28S) HKGs were evaluated using real-time PCR and were compared in different physiological stages of the endometrium. Endometrial biopsies were obtained from mares on day of ovulation (d0, n = 4), at late diestrus (LD, n = 4), after luteolyis (AL, n = 4) of the cycle and on days 14 (P14; n = 3), 18 (P18, n = 3) and 22 (P22; n = 3) of pregnancy. A model based on REML with support of descriptive statistics was proposed in accordance with experimental design and was further confirmed with principal component analysis (PCA). Results were compared with widely used software including geNorm, BestKeeper, and NormFinder. Results indicated that GAPDH was the most stable HKG and RPL32 was ranked as the second best. 18S and 285 were found to be the least stable. The proposed model, PCA, geNorm, and BestKeeper were in agreement in detecting the most stable and the least stable HKGs in the equine endometrium during the estrous cycle and early pregnancy. (C) 2011 Elsevier B.V. All rights reserved.
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    Toll-like receptor 2 and 4 expression in the bovine corpus luteum during the different stages of the estrous cycle
    (Brazilian Coll Animal Reproduction, 2017) Atli, Mehmet Osman; Kose, Mehmet; Kaya, Mehmet Salih; Aydilek, Nurettin; Guzeloglu, Aydin; Wiltbank, Milo C.
    The aim of this study was to elucidate the presence of components of the innate immune system in the bovine corpus luteum (CL) by detecting the expression and cell-specific localization of TLR2 and TLR4 during different stages of the estrous cycle in a control study design. Bovine CL samples were collected from a local slaughterhouse and assigned to three groups as follows: developing CL (dCL; n = 6, approx. days 3-6), mature CL (mCL; n = 5, approx. days 8-12), and regressing CL (rCL; n = 5, approx. days 17-19). An upregulation of TLR2 mRNA was detected only in rCL (P < 0.05). Localization of the TLR2 protein was particularly apparent in luteal cells and a prominent immunofluorescent signal corresponding to TLR2 was detected only in rCL. TLR4 mRNA were higher in mCL and rCL compared to dCL (P < 0.05). The presence of the TLR4 protein in bovine CL was clearly detected in the luteal cells of both mCL and rCL. The results of this study suggest a role for TLRs in the development, maintenance, and regression of bovine CL. TLR signaling mediated pathway in luteal cells may involve in the regression of CL via regulation of TLR2 and TLR4.