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Purpose To compare the central corneal thickness (CCT), intraocular pressure (IOP), and tear insulin-like growth factor 1 (IGF-1) levels between patients with acromegaly and a control group and to evaluate the possible effect of tear IGF-1 and duration of the disease on CCT and IOP. Methods We included 31 patients with acromegaly (study group) and 40 age- and sex-matched controls in the study. Patients with acromegaly were divided into 2 subgroups based on disease status (active/inactive). All participants underwent complete ophthalmologic evaluation including CCT and IOP values. Basal tear samples were collected from both groups and tear IGF-1 levels were measured. The CCT, IOP, and tear IGF-1 levels were compared between groups and subgroups and the association between tear IGF-I levels and ocular parameters (CCT, IOP) and disease duration were also evaluated. Results Central corneal thickness, IOP, and tear IGF-1 levels did not show a significant difference between study and control groups. We also did not find a significant difference in terms of CCT, IOP, or tear IGF-1 levels between subgroups of patients. Correlation analysis did not show an association between the duration of disease and tear IGF-1 levels with CCT or IOP. Conclusions There was no significant difference in tear IGF-1 levels between patients with acromegaly and controls. Additionally, there was no correlation between disease duration and tear IGF-1 levels with CCT or IOP levels. This lack of association may suggest that tear IGF-1 levels might not have an effect on CCT or IOP findings in patients with acromegaly.

WOS: 000383902000015 PubMed: 27435174 Aim: Contrast medium-induced nephropathy is one of the major complications of intravenous contrast medium use. But its pathogenesis is unclear. Epithelial mesenchymal transition (EMT) is defined as the transformation of the primer epithelial cells to mesenchymal cells. EMT in tubular cells might cause tubulointerstitial damage. In this study, we investigated whether or not EMT has a role in radiocontrast-induced nephropathy. Radiocontrast medium might be triggering reversible EMT via serum and glucocorticoid-regulated kinase 1 (SGK 1). We investigated the effect of different concentrations of the contrast agent iopromide on human proximal tubule cell (HK-2) culture by measuring the level of SGK1, snail family zinc finger 1 (SNAIL1), connective tissue growth factor (CTGF), and collagen type I alpha 1 (COL1A1).Methods: We conducted a scratch assay and qPCR. HK-2 cells were cultured in the petri dishes/flasks and starved with serum-free medium. The 40, 20, and 10mg/mL doses of iopromide were administrated to cells. The scratches were photographed immediately and again at the 20th hour. The levels of gene expression of SGK1, SNAIL1, CTGF, and COL1A1 were measured using the real-time qPCR system at the end of the 24th hour.Results: Iopromide caused the breaking of intercellular connections, the disappearance of the cobblestone appearance of cells, and the migration of cells at the 20th hour in the scratch assay. It also increased the expression of SGK1, SNAIL1, CTGF, and COL1A1 genes.Conclusion: Our study concluded that certain important markers of EMT increase in different concentrations of the contrast agent. High osmolality might trigger EMT. The relationship between contrast agent and EMT has not been defined before. Further in vivo and in vitro studies are required. research foundation of Ondokuz Mayis University [PYO.TIP.1901.13.046] This study was supported by the research foundation of Ondokuz Mayis University [PYO.TIP.1901.13.046].

WOS: 000374903500025 PubMed: 26419593 Metastasis occurs due to migration of the cells from primary tumor toward other tissues by gaining invasive properties. Since metastatic invasion shows a strong resistance against conventional cancer treatments, the studies on this issue have been focused. Within this context, inhibition of migration and determination of the relationships at the gene level will contribute to treatment of metastatic cancer cases. We have aimed to demonstrate the impact of TGF-beta 1 and fluvastatin on human breast cancer (MCF-7) and human hepatocellular carcinoma (Hep3B) cell cultures via Real-Time Cell Analyzer (RTCA) and to test the expression levels of some genes (NDRG1, SGK1, TWIST1, AMPKA2) and to compare their gene expression levels according to RTCA results. Both of cell series were applied TGF-beta(1) and combinations of TGF-beta(1)/fluvastatin. Primer and probes were synthesized using Universal Probe Library (UPL, Roche) software, and expression levels of genes were tested via qPCR using the device LightCycler 480 II (Roche). Consequently, fluvastatin dose-dependently inhibited migration induced by TGF-beta 1 in both groups. This inhibition was accompanied by low level of SGK1 messenger RNA (mRNA) and high levels of NDRG1 and AMPKA2 mRNA. Thus, we conclude that fluvastatin plays an important role in reducing resistance to chemotherapeutics and preventing metastasis.

ozkaya, ozan/0000-0002-0198-1221 WOS: 000340259600020 PubMed: 24845105 Cisplatin is one of the commonly used anticancer drugs and nephrotoxicity limits its use. The aim of this study is to investigate the possible protective effect of creatine supplementation on cisplatin-induced nephrotoxicity. Sixty male Sprague-Dawley rats were divided into three groups: Group I: Cisplatin (n = 20) (7 mg/kg cisplatin intraperitoneal (i.p.) single dose), group II: Cisplatin + creatine monohydrate (n = 20) (7 mg/kg cisplatin i.p. single dose and 300 mg/kg creatine p.o. daily for 30 days starting on first day of cisplatin injection), group III: Control group (n = 20) (Serum physiologic, 2.5 mL/kg i.p.). Sacrifications were performed at first week and 30th day. Blood urea nitrogen (BUN) and serum creatinine levels, histopathological evaluation, mitochondrial deoxyribonucleic acid (mtDNA) common deletion rates, and body weights of rats were evaluated. A significant decrease in body weight, higher values of kidney function tests, histopathological scores, and mtDNA deletion ratios were observed in group I compared to control group at days 7 and 30 (p

Guvenc, Tolga/0000-0003-1468-3415 WOS: 000332484300008 PubMed: 24122261 Aspiration pneumonitis refers to acute chemical lung injury caused by aspiration of sterile gastric contents. The aim of this study was to evaluate the role of quercetin (QC) in acid aspiration-induced lung injury in rats. Twenty-eight female Sprague-Dawley rats were used and divided into the following groups (n = 7): sham (aspirated normal saline, S), hydrochloric acid (aspirated HCl), S plus treatment with QC (S + QC), and HCl plus treatment with QC (HCl + QC). After aspiration, the treatment groups received QC 60 mg/kg/day intraperitoneally once a day for 7 days. As a result of acid aspiration, an increase was observed in the levels of serum clara cell protein-16 (CC-16) and advanced oxidation protein products, whereas there was a decrease in serum thiobarbituric acid-reactive substances, superoxide dismutase (SOD), and catalase levels. There was a significant decrease in peribronchial inflammatory cell infiltration, alveolar septal infiltration, alveolar edema, and alveolar exudate scores, except in the alveolar histiocytes in the HCl + QC group. The expression of nitric oxide synthase, which increased after aspiration in the HCl group, showed a statistically significant decrease after the QC treatment. After the treatment with QC, an increase in the serum SOD level was observed, whereas a significant decrease was determined in the serum CC-16 level relative to that of the aspiration group (HCl). The antioxidant QC is effective in the treatment of lung injury following acid aspiration and can be used as a serum CC-16 biomarker in predicting the severity of oxidative lung injury.

OZET Roziglitazon; Tip II diabet tedavisinde kullanilan tiazolidinedion grubu bir ilactir. Bu grup ilaclar, karbonhidrat ve lipid metabolizmasini kontrol eden genlerin ekspresyonu¬nun duzenlenmesinde gorev alan peroksizom proliferator aktive reseptor-gama (PPAR-gama)’nin ligandi olarak rol oynarlar. Telomer; kromozom uclarinda bulunan ozellesmis DNA ve protein kompleksinden olusmaktadir. Omurgalilarda telomerik DNA, TTAGGG gibi, guaninden zengin dizelerden olusur. Normal hucrelerde, her DNA replikasyon per¬yodunda, telomer 50–150 baz kisalir. Tamir edilemeyen oksidatif DNA hasari da bu te¬lomer kisalmasina katkida bulunabilir. Literaturde, roziglitazonun telomer uzunluguna etkisini gosteren bir calisma bulunmamaktadir. Bu nedenle, calismamizda roziglitazonun telomer uzunluguna etkisini arastirmayi amacladik. Calismada 7–8 haftalik, 248-275 g arasinda, 16 tane erkek Sprague-Dawley cinsi rat kullanildi. Ratlar; her grupta dort tane olacak sekilde dort gruba ayrildi ve 14 gun boyunca 0, 0,5, 1,0 ve 2,0 mg/kg dozlarinda roziglitazon oral yoldan uygulandi. Rat karaciger, ince bagirsak, pankreas dokulari ve lenfositlerinden genomik DNA izolasyonu yapildi. Numunelerde genom sayisini belir¬leyebilmek icin tek kopya gen olarak 36B4 geni secildi. 36B4 genine spesifik primerle¬rin kullanildigi Q-PCR ile bir rat dokusundaki genom kopya sayisi bulundu. Rat genom standardi olarak kullanilan bu numune tum numunelerdeki genom sayisini bulmak icin kullanildi. Tum numunelerde telomer (T) ve 36B4 (S) icin kantitatif PCR yapildi. Iki ca¬lismada ile elde edilen urunler birbirine oranlarak her numune icin T/S orani daha sonra da bir kalibrator numune kullanilarak relatif T/S orani hesaplandi. Ayrica 10,2 kb telo¬mer standardi kullanarak numunelerin telomer uzunluklari kilobaz olarak da hesaplandi. Farkli roziglitazon dozlari verilen numuneleri kontrol grublari ile karsilastirildigimizda telomer uzunlugu acisindan istatistiksel olarak anlamli bir fark bulamadik (p>0,05). Bu sonuc, roziglitazonun farkli rat dokularinda telomer kisalmasina yol acmadigini gostermektedir. Research on the effect of rosiglitazon on telomere dynamics with molecular methods in rats ABSTRACT Rosiglitazone that is a drug for the treatment of type II diabetes mellitus, belong to the class of thiazolidinediones. Drugs of this class act as ligands for the peroxisome prolif¬erator-activated receptor-gamma (PPAR-gamma), which is involved in the regulation of genes controlling carbohydrate and lipid metabolism. Telomeres are composed of spe¬cialized DNA and protein complexes. Telomeric DNA consist of guanine-rich sequences, such as TTAGGG in vertebrates. In normal cells, telomeres shorten by ~50–150 bp with each round of DNA replication. In addition to this, unrepaired oxidative DNA damage can also contribute to telomere shortening. There is no study that shows the effect of rosiglitazone on telomere length in literature. Because of this, we aimed to research on the effect of rosiglitazone on the telomere length in our study. We used sixteen male Sprague- Dawley rats that at 7-8 weeks of age and weighing 248–275 g in our study. The rats were divided into four groups, four rats in each group, and dosed once a day for 14 days by oral gavage with doses of 0.0, 0.5, 1.0, and 2.0 mg/kg rosiglitazone. Genomic DNA was extracted from rat liver, small intestine, pancreas tissue and lymphocytes. The 36B4 genewas chosen as single copy gene for determine the number of genom in the samples. The number of genome copies were found in a rat tissue by qPCR that is used the 36B4 gene-specific primers. This sample, that is used as rat genom standarts, was used for find the number of genome in all samples. The qPCR was performed for telomere (T) and 36B4 (S) in all samples. The T / S ratio was calculated by the ratio of the products derived from two studies to each other, and then the relative T / S ratio was calculated by using a calibrator sample for each sample. In addition, telomere lengths of the samples were also calculated as kilobase by using 10,2 kb long telomere standard. We did not found statistically any significant difference between doses of rosiglitazone and control groups for telomere length (p>0.05). This result has showed that rosiglitazone doesn’t lead to telomere shortening. J. Exp. Clin. Med., 2011; 28:162-167

Guven, Mustafa/0000-0001-8643-9775; WOS: 000347840000038 PubMed: 25790559 Background/aim: Brain ischemia and treatment are important topics in neurological science. Free oxygen radicals and inflammation formed after ischemia are accepted as the most significant causes of damage. Currently there are studies on many chemopreventive agents to prevent cerebral ischemia damage. Our aim is to research the preventive effect of the active ingredient in syringic acid, previously unstudied, on oxidative damage in cerebral ischemia. Materials and methods: The rats were randomly divided into 4 groups: control group (no medication or surgical procedure), sham group (artery occlusion), artery occlusion + syringic acid group sacrificed at 6 h, and artery occlusion + syringic acid group sacrificed at 24 h. Obtained brain tissue from the right hemisphere was investigated histopathologically and for tissue biochemistry. Results: Superoxide dismutase and nuclear respiratory factor 1 values decreased after ischemia and they increased after syringic acid treatment, while increased malondialdehyde levels after ischemia were reduced after treatment. Caspase-3 and caspase-9 values increased after ischemia and decreased after treatment; this reduction was more pronounced at 24 h. Conclusion: Our study revealed that syringic acid treatment in cerebral ischemia reduced oxidative stress and neuronal degeneration. In the light of the biochemical and histopathologic results of the present study, we think that syringic acid treatment may be an alternative treatment method.

WOS: 000344075800003 PubMed: 25335733 N-myc downstream-regulated gene 1 (NDRG1) is defined as metastasis suppressor and can be downregulated in many types of cancers, and reported to be an indication of tumor progression in hepatocellular carcinomas. Several in-vivo and in-vitro studies have demonstrated that iron chelators such as Desferrioxamine (DFO) and 1-10 Phenanthroline (PHEN) are effective antitumor agents. It is suggested that these chelators deliver their antitumor activity by acting on the NDRG1 gene expression. It remains unclear why NDRG1 gene expression affects the tumors differently, or becomes affected differently. We consider that this different effect might be caused by variants. Based on this information, we developed specific primers and probes for NDRG1 mRNA variants using bioinformatics analysis, and investigated how DFO and PHEN affected the dynamics of NDRG1 variant on the cell lines of Human Breast Adenocarcinoma (MCF-7) and Hepatocellular Carcinoma (HepG2) that demonstrate opposite action for the relationship NDRG1-metastasis. We administrated various doses of DFO and PHEN into the cells to monitor cell vitality and proliferation with Real time Cell Analyzer. We analyzed the gene expression levels of study groups with Quantitative RT-PCR as well as relative gene expression. Variants of NDRG1 mRNA were transcriptionally regulated after HepG2 and MCF-7 cells were treated by iron chelators, resulting in domination of NDRG1 mRNA Variant 1 (V1) in the HepG2 calls and domination of NDRG1 mRNA Variant 2 (V2) in the MCF-7 cells. Anti-proliferative and cytotoxic effects were observed in the MCF-7 cells whereas an increased proliferation was present in the HepG2 cells. Ondokuz Mayis UniversityOndokuz Mayis University; [OMU PYO. TIP.1904.11.028.26] This study was supported by Ondokuz Mayis University. The Project number was OMU PYO. TIP.1904.11.028.26.

AYGUN, CANAN/0000-0002-7955-5943 WOS: 000339860800004 PubMed: 25008013 Background: Previous studies have shown the relationship between in utero lung development and vitamin D [25(OH)D], but there have been no studies to investigate whether vitamin D deficiency is a risk factor for respiratory distress syndrome (RDS) in preterm babies. Objectives: In this study, we investigated if 25(OH)D deficiency is a risk factor for RDS. Methods: One hundred fifty-two preterm newborns, born at 29-35 weeks gestational age, were included in the study following informed consent from the parents. Peripheral blood samples were collected within the first 24 hours of life and 25(OH)D levels were measured by liquid chromatography-tandem mass spectrometry. Demographic characteristics of the babies and the diagnosis of RDS were recorded. Results: In 64% of preterm infants, 25(OH)D levels were compatible with severe deficiency (

Background: A nationwide multicenter study was organized to establish reference intervals (RIs) in the Turkish population for 25 commonly tested biochemical analytes and to explore sources of variation in reference values, including regionality. Methods: Blood samples were collected nationwide in 28 laboratories from the seven regions (?400 samples/region, 3066 in all). The sera were collectively analyzed in Uludag University in Bursa using Abbott reagents and analyzer. Reference materials were used for standardization of test results. After secondary exclusion using the latent abnormal values exclusion method, RIs were derived by a parametric method employing the modified Box-Cox formula and compared with the RIs by the non-parametric method. Three-level nested ANOVA was used to evaluate variations among sexes, ages and regions. Associations between test results and age, body mass index (BMI) and region were determined by multiple regression analysis (MRA). Results: By ANOVA, differences of reference values among seven regions were significant in none of the 25 analytes. Significant sex-related and age-related differences were observed for 10 and seven analytes, respectively. MRA revealed BMI-related changes in results for uric acid, glucose, triglycerides, high-density lipoprotein (HDL)-cholesterol, alanine aminotransferase, and ?-glutamyltransferase. Their RIs were thus derived by applying stricter criteria excluding individuals with BMI >28 kg/m2. Ranges of RIs by non-parametric method were wider than those by parametric method especially for those analytes affected by BMI. Conclusions: With the lack of regional differences and the well-standardized status of test results, the RIs derived from this nationwide study can be used for the entire Turkish population. © by De Gruyter 2014., Applied Scientific Research Fund: :24256003:2012-2014 Japan Society for the Promotion of Science, Acknowledgments: This study was supported by Research Fund of Uludag Universty (UAP(T)-2011/48), IFCC C-RIDL, Scientific Research Fund (No:24256003:2012-2014) provided by the Japan Society for the Promotion of Science, Abbott Diagnostics and Becton Dickinson. We had invaluable input from the IFCC C-RIDL and the Turkish National Biochemical Association. We are very grateful to Uludag University Central Laboratory, We are thankful to David Ambruster from Abbott Diagnostics for his kind cooperation to realize this study. The final manuscript was edited with generous support by Prof. A. Myron Johnson.

WOS: 000344349600003 PubMed: 24505052 The main purpose of this study was to assess the role of S100B protein, neuron-specific enolase (NSE), and glial fibrillary acidic protein (GFAP) in the evaluation of hypoxic brain injury in acute carbon monoxide (CO)-poisoned patients. This cross-sectional study was conducted among the patients with acute CO poisoning who referred to the emergency department in a 1-year period. Serum levels of S100B protein, NSE, and GFAP were determined on admission. A total of 55 CO-poisoned patients (mean age +/- standard deviation, 45 +/- 20.3 years; 60% women) were included in the study. The control group consisted of 25 healthy adults. The patients were divided into two groups according to whether they were conscious or unconscious. The serum levels of S100B, NSE, and GFAP were higher in patients than that in the control group. There was no significant difference between unconscious and conscious patients with respect to these markers. There was a statistically significant difference between the conscious and unconscious patients and the control group in terms of S100B and NSE levels. There was also a statistically significant difference between the unconscious patients and the control group in terms of GFAP levels. Increased serum S100B, NSE, and GFAP levels are associated with acute CO poisoning. These biomarkers can be useful in assessing the clinical status of patients with CO poisoning. Scientific Research Council of Ondokuz Mayis UniversityOndokuz Mayis University [PYO.TIP.1901.12.036] This study was supported by the Scientific Research Council of Ondokuz Mayis University with the number of PYO.TIP.1901.12.036.

WOS: 000337094900112 PubMed: 24622883 Histone deacetylase (HDAC) inhibitors, such as trichostatin A (TSA), and iron chelators, including deferoxamine (DFO) and phenanthroline (PHEN), appear to have anticancer effects. We hypothesized that the HDAC inhibitors and iron chelators would be synergistic with their effect on breast cancer cell line MCF7, because the HDAC inhibitors increase glucose-regulated protein 78 (Grp78) and the iron chelators reduce its expression. Although the administration of TSA alone resulted in a dose-related decrease in the cell index, it did not have an antiproliferative effect except the 62.5 and 500 nM of TSA. However, all doses of TSA produced a cytotoxic effect from the initial hours when combined with 150 mu M of DFO and 25 mu M of PHEN. DFO and PHEN downregulated Grp78, Grp94, and MRP1 expressions and upregulated CHOP and HO-1 expressions. TSA upregulated all the genes in various rates when used alone but resulted in decreased expression levels when combined with DFO and PHEN. Increased HDAC-1 levels in the Grp78 promoter region indicated that DFO and PHEN either promoted binding of HDAC-1 to this region or inhibited its detachment. We determined that the reduction of increased Grp78, Grp94, HO-1, and MRP1 expressions, which appears to inhibit the chemotherapeutic effect of TSA, through the combination with DFO or PHEN will contribute to the anticancer effect. Ondokuz Mayis UniversityOndokuz Mayis University; [PYO.TIP. 1901 09 001] This study was supported by the Ondokuz Mayis University. The project number was PYO.TIP. 1901 09 001.

WOS: 000347538100004 PubMed: 25347557 Fluvastatin (FLU) prevents the conversion of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) to mevalonic acid by inhibiting HMG-CoA reductase and decreases cholesterol level. Although the effects of FLU treatment on several cancer types through many mechanisms have been identified, its relationship with unfolded protein response and apoptosis has not been clearly understood. In this recent study, we aimed to investigate the cytotoxic effect of Fluvastatin on MCF-7 cells and define the transcriptional regulation of specific genes during the occurrence of this cytotoxic effect. We administered 0.62, 2.5, 5, and 40 mu M FLU on MCF-7 cells singly and in combination with 2-deoxyglucose (2-DG), and we monitored cell viability and proliferation for 48 hours using real-time cell analyzer system (xCELLigence). At the same time, we measured the mRNA expression levels of glucose-regulated protein 78 (GRP78), CCAAT/enhancer binding protein, homologous protein (CHOP), caveolin-1 (CAV1), NDRG1 Variant 1 and Variant 2, HMOX1, SGK1, and prostate apoptosis response-4 (PAR4) genes using quantitative real-time polymerase chain reaction (LightCycler 480 II). We accepted GAPDH gene and control groups as the reference gene and calibrator, respectively. We performed relative gene expression analyses of the study groups using the QIAGEN 2009 Relative Expression Software Tool (REST). FLU revealed an antiproliferative and cytotoxic effect on MCF-7 cells, while causing the transcriptional regulation of many genes. Of these genes, the mRNA expressions of CHOP, heme oxygenase 1 (HMOX1), N-myc downstream-regulated gene 1 (NDRG1) V1, and NDRG1 V2 increased. On the other hand, the mRNA expression levels of SGK1 and CAV1 decreased. The antiproliferative effects of FLU may be related to the decreased expression levels of SGK1 and CAV1.

Yuksel, Serkan/0000-0001-9501-4568 WOS: 000325413800004 PubMed: 23301552 Hypertension terms "dipper" and "non-dipper" are propounded by the change that occurs during ambulatory blood pressure (BP) monitoring. The purpose of this study is to present whether the serum urotensin II levels are different in patients with dipper and non-dipper hypertension and to put forward the effects causing this difference, if there are any. Patients recently diagnosed with hypertension were included in the study. With ambulatory BP monitoring, 81 patients with high BP were divided into two groups, dipper (n = 40) and non-dipper (n = 41). Serum urotensin II levels were analyzed by ELISA method. Serum urotensin II levels were higher in patients with non-dipper hypertension than in patients with dipper hypertension (204 [106-533] vs. 140 [96-309], P = .004). There was a positive correlation between total systolic BP and serum urotensin II levels (r = 0.408 and P = .009), but the relation in the non-dipper hypertension group was not significant (r = 0.194 and P = .2). In conclusion, serum urotensin II levels were higher in non-dipper HT patients than dipper HT patients. This higher urotensin II level might be responsible for poor prognoses.

Caprinuloidea rudists are locally abundant and widespread in Lower Cretaceous (Albian Stage) Edwards Formation in Texas. Landward of the shelf margin on the shallow marine Comanche Shelf rudists built circular and elongate bioherms with coarse-grained flank deposits. Two caprinid morphotypes suggest that some lived as elevators above the substrate and others were recumbent upon mobile grain flats. Elevators have elongated attached valves and weakly coiled free valves and recumbents have arcuate attached valves and strongly coiled free valves. Detailed morphologic studies are not possible on the many molds and casts, but a few specimens are silicified. Their internal structures can be seen by X-ray computed tomographic scanning (CT), which provides three-dimensional representations of internal features. This technique enables the specific identification of caprinid rudists that otherwise could only be identified by sectioning the specimen. The abundant Edwards species is identified as Caprinuloidea perfecta because it has only two rows of polygonal canals on its ventral and anterior margins. X-ray CT images reveal ontogenetic stages of these unusual gregarious bivalves. Allometric to isometric growth characterizes the left-free valve (LV). Although the prodissoconch is unknown, the plots suggest that the initial length was greater than the width, which is like the D-shaped prodissoconch of Cardiacea. The LV has the morphology of loosely coiled gastropods and the right-attached valves are elongated and are unlike most Bivalvia.

Hucre icerisinde bir cok onemli gorevi bulunan endoplazmik retikulumun (ER) bir diger onemli gorevi de protein katlanmasinin saglanmasidir. ER’de protein katlanmasinin gerceklesmesini saglayan glukoz duzenleyici protein 78, glukoz duzenleyici protein 94, lektin benzeri proteinler ve foldazlar bulunmaktadir. ER fonksiyon kapasitesini asan fizyolojik veya patolojik durumlarda lumende katlanmamis ya da yanlis katlanmis protein birikimi meydana gelir ve bu durum ER stresi olarak isimlendirilmektedir. ER stresi hucre icerisinde olustuktan sonra hucrenin hayati fonksiyonlarini devam ettirebilmek icin katlanmamis protein cevabi (UPR) adi verilen sinyal yolaklari devreye girmektedir. Protein kinaz RNA (PKR) benzeri ER kinaz (PERK), aktive edici transkripsiyon faktoru (ATF6), inozitol gerektiren kinaz 1 (IRE1) yolaklari UPR sinyal yolaklarinin baslicalaridir. Endoplasmic reticulum stress and unfolded protein response Another important function of the endoplasmic reticulum (ER), which has many important tasks in the cell, is to ensure the protein folding. In ER, there are glucose regulated protein 78, glucose regulated protein 94, lectin like proteins and foldases, which enable the protein folding. In case of physiological and pathological situations that exceed the function capacity of ER, unfolded or misfolded protein accumulation occurs in lumen and this case is named as ER stress. After the ER stress occurs in the cell, the signal pathways named unfolded protein response (UPR) step in for the continuance of the vital functions of the cell. Major UPR signal pathways include protein kinase RNA (PKR) like ER kinase (PERK), activating transcription factor 6 (ATF6), j requiring kinase 1 (IRE1). J. Exp. Clin. Med., 2012; 29:95-100

OZET Telomer, kromozom uclarinda TTAGGG gibi, guaninden zengin kisa dizelerin ardisik tekrarlarindan ve buna bagli bazi proteinlerden olusan bir kompleksdir. Farkli turdeki canlilarda ve ayni turde farkli hucre ve dokularda farkli uzunluklara sahiptir. DNA replikasyonu esnasinda telomer 50-150 baz kadar kisalarak kromozomal stabilitenin korunmasina katkida bulunur. Telomerin yaslanma, kanser ve normal hucre biyolojisindeki onemini anlayabilmek icin yapilan calismalarda; telomer uzunlugu, altin standart olarak kabul edilen Southern Blot yaninda birkac farkli metodla olculmustur. Bu metodlarin kendine gore ustun ve eksik yonleri bulunmaktadir. Biz de bu calismada, ratlarin telomer uzunlugu olcumunde kullanabilecegimiz ve daha sonraki calismalara da uygulanabilir bir kantitatif PCR protokolu olusturmayi ve bu protokolle farkli rat dokularindaki telomer uzunluklari hakkinda bilgi edinmeyi amacladik. Dort erkek Sprague-Dawley cinsi ratin karaciger, ince bagirsak ve pankreas dokusu ve lenfositlerinden DNA izolasyonu yapildi. Numunelerde genom sayisini belirleyebilmek icin tek kopya gen olarak 36B4 geni secildi. 36B4 genine spesifik primerlerin kullanildigi Q-PCR ile bir rat dokusundaki genom kopya sayisi bulundu. Rat genom standardi olarak kullanilan bu numune tum numunelerdeki genom sayisini bulmak icin kullanildi. Tum numunelerde telomer (T) ve 36B4 (S) icin kantitatif PCR yapildi. Iki calismada ile elde edilen urunler birbirine oranlarak her numune icin T/S orani daha sonra da bir kalibrator numune kullanilarak relatif T/S orani hesaplandi. Ayrica 10,2 kb telomer standardi kullanarak numunelerin telomer uzunluklari kilobaz olarak da hesaplandi. Relatif kantitasyon icin mutlak gerekli olan Telomer ve 36B4 PCR calismalarinda % 100 verimlilige ulastik. Bunu logaritmik baslangic konsantrasyona karsi ΔCT grafigi slope degerinin 0,1’den kucuk olmasi ile de gosterdik. Ayrica telomer uzunlugu acisindan rat dokulari arasinda anlamli bir fark bulundu (p < 0,05). Sonuc olarak rat dokularinda telomer uzunlugu olcumu icin hassas, kolay ve hizli bir Q-PCR protokolu olusturdugumuzu dusunmekteyiz. The measurement of telomere length in sprague-dawley rats by quantitative PCR ABSTRACT Telomere consist of sequential repetitions of guanine-rich short sequences, such as TTAGGG, and some related proteins complexes in the ends of chromosomes. The dif¬ferent organisms and different cells and tissues of same organisms have different telom¬ere lengths. Telomere contributes to the protection of chromosomal stability by shorten ~50–150 base during DNA replication. Telomere length was measured by several differ¬ent methods as well as Southern blot, which is considered the gold standard in the studies to understand the importance of telomere in aging, cancer and normal cell biology. These methods has specific superior and limitations to himself. In this study, we aimed to create a quantitative PCR protocol, we can use to measurement of telomere length of rats and can also be applied in subsequent studies, and to obtain information about the telomere lengths in the different tissues of rats by this protocol. Genomic DNA was extracted from liver, small intestine, pancreas tissue and lymphocytes of four male Sprague-Dawley rat. The 36B4 gene was chosen as single copy gene for determine the number of genom in the samples. The number of genome copies were found in a rat tissue by qPCR that is used the 36B4 gene-specific primers. This sample, that is used as rat genom standarts, was used for find the number of genome in all samples. The qPCR was performed for telomere (T) and 36B4 (S) in all samples. The T / S ratio was calculated by the ratio of the products derived from two studies to each other, and then the relative T / S ratio was calculated by using a calibrator sample for each sample. In addition, telomere lengths of the samples were also calculated as kilobase by using 10,2 kb long telomere standard. We reached to 100% of efficiency in the studies of telomere and 36B4 PCR, it is necessary to the relative quantitation. We also demonstrated by the value of slope in ΔCt graphics versus logaritmic initial concentration is less than 0,1. In addition, a statistically any significant difference found between tissues of rat for telomere length (p