Band 44, Heft 4

Objectives Since studies regarding the effect of polysomy 17 (P17) in breast cancer cases with some specific clinical findings are few in number and are in small sample sizes, meta-analysis was implemented to exhibit the effects of P17 in patients with Human Epidermal growth factor Receptor 2 (HER2) amplification on lymph node involvement and tumor grade. Materials and methods Pubmed literature database was scanned up to June 2017 by using the keywords “polysomy 17 breast cancer” and 141 studies were accessed. Ultimately four of the reviewed papers have been found to be appropriate for examining the effect of P17 on lymph node involvement and tumor grade. Prior to meta-analysis, publication bias and heterogeneity of the studies was examined. Results Meta-analysis in the examining the effect of polysomy 17 on lymph node involvement (OR=1.708, 95% CI: 1.068–2.733), on grade [3]/[3,1] (OR=3.402, 95% CI: 1.726–6.707), on grade [3]/[3, 2] (OR=2.581, 95% CI: 0.778–8.559) and on grade [2]/[2,1] (OR=1.854, 95% CI: 0.531–6.468) was determined in those with HER2 amplification. Conclusion It was observed that in terms of lymph node involvement, P17 was a risk factor in patients and with regard to tumor grade, P17 was a risk factor when grade increased in patients with amplification.

Background In this study, the objective was to evaluate the diagnostic performance of some miRNAs, which were shown to have a diagnostic value for prostate cancer (PCa), and the effect of chronic prostatitis in distinguishing benign prostatic hyperplasia (BPH) and PCa. Materials and methods Serum levels of 11 miRNAs were investigated in BPH, chronic prostatitis and PCa patients. Measurements were performed using qRT-PCR. Results In the analysis, serum levels of miR-375, -125b-5p, -30c-5p, -26b-5p, and let-7c-5p were downregulated in cancer compared with non-cancer group and AUCs of these miRNAs in distinguishing PCa group from non-cancer group were calculated as 0.781, 0.782, 0.762, 0.874, and 0.845, respectively. AUC of the combination of miR-375 and miR-26b-5p in distinguishing PCa group from non-cancer group was 0.891, AUC of these two miRNAs in distinguishing PCa group from BPH group was 0.944. Conclusion In our study, 11 miRNAs were studied and 5 of these miRNAs were considered as biomarker candidates as these miRNAs, individually or combined, could be used to distinguish PCa from benign conditions. Furthermore, a higher specificity and sensitivity were obtained in distinguishing BPH and PCa when data for diagnostic potential of miRNAs were analyzed without including chronic prostatic group.

Background Cytotoxic, antiproliferative, cell cycle inhibitive, oxidative and apoptotic effects of cetuximab [antibody for epidermal growth factor receptor (EGFR)] alone and together with stabilized silver ion solution (St-Ag) on P-H1299, R-H1299, A-431 and A-549 cells were investigated. Materials and methods Cytotoxic effects of cetuximab alone and together with St-Ag on cells were determined by Cell Titer-Blue ® Cell Viability and Lactate Dehydrogenase Activity tests. Cell cycle distributions and apoptosis were detected by reverse transcription polymerase chain reaction (RT-PCR). Results St-Ag enhanced cetuximab cytotoxic effect on all cells. LDH activity, as a result of cell death, was found the highest level at treatment of cetuximab with St-Ag in all cells. Both treatment increased caspase-3/7 activity which is apoptotic enzyme was found higher in A-549 cells than other cells. Also, treatment of cetuximab with St-Ag caused increasing Bax/Bcl-2 ratio in all cells. Cetuximab with St-Ag treatment increased glutathione peroxidase activity in all cells generating oxidative stress. Proliferating Cell Nuclear Antigen (PCNA), topoisomerase II-alpha (except R-H1299), cyclin D1 and D2 genes expression were decreased in all cells which explain the cell cycle inhibition effect. Conclusion These findings suggest that treatment of cetuximab combined with St-Ag exhibit more carcinogenesis reducing potential than cetuximab alone.

Objective Cancer Antigen 125 (CA125) and Risk of Ovarian Malignancy Algorithm (ROMA) score are used for classification of ovarian masses (benign/malign) in preoperative stage. However, their discrimination capacity are considered insufficient, and greatly effected by histological subtype and menopausal status. This study aimed to investigate diagnostic performance of Human epididymis protein 4 (HE4), Y (tyrosine), K (lysine), and L (leucine)-40 (YKL-40), Mesothelin, Rho GDP dissociation inhibitor ß (LyGDI), CA125 or their combinations in discrimination of benign/malign ovarian diseases in preoperative stage. Materials and methods The study groups were comprised sera of 31 epithelial ovarian cancer (EOC), 30 benign ovarian tumor patients, and 32 healthy women. The diagnostic performance of the biomarkers were evaluated based on ROC-AUC values and logistic regression analysis incorporating menopausal status and clinical diagnosis of the subjects. Results Our data demonstrates that “CA125-HE4-Mesothelin-YKL-40” had the highest sensitivity at 80%, 90%, 95% specificity 96.8%, 93.6%, 93.6%, respectively. Conclusion This study provides the first evidence for the combinational uses of “CA125-HE4-Mesothelin-YKL-40” as a panel in distinguishing malign from benign ovarian tumor, not affected by menopausal status unlike ROMA. However, higher patient number may also provide the evaluation of this panel in malign group in terms of tumor stages.

Objective This study aimed to investigate the renoprotective effects of paricalcitol, a synhetic vitamin D analog, through its possible roles on p38 MAPK and PI3K/Akt signaling pathways to prevent oxidative stress, inflammation and apoptosis during renal I/R. Materials and methods Total 20 kidney tissues of sham (n = 6), subjected to renal I/R bilaterally for 45 min ischemia followed by 24 h reperfusion (n = 7) and paricalcitol (0.3 μg/kg, ip) pretreated Wistar albino rats (n =7) were used in this study. Interstitial inflammation and active caspase-3 expression were evaluated histologically. TNF-α, IL-1β, kidney injury molecule-1 (KIM-1), MDA and SOD activity in kidneys were analysed biochemically. Furthermore, activation of p38 MAPK, PI3K/Akt signaling pathways and NFκB p65 were evaluated by western blot. Results Paricalcitol pretreatment significantly reduced interstitial inflammation during renal I/R, which was consistent with decreased tumor TNF-α, IL-1β, active caspase-3 and KIM-1 expression. Paricalcitol also reduced MDA level and attenuated the reduction of SOD activity in the kidney during I/R. Moreover, paricalcitol could suppress the p38 MAPK and NFκB p65, and also activate PI3K/Akt signaling pathway during renal I/R. Conclusion All these findings indicate that paricalcitol may be an effective practical strategy to prevent renal I/R injury.

Background Cytoplasmic sialidase (NEU2) plays an active role in removing sialic acids from oligosaccharides, glycopeptides, and gangliosides in mammalian cells. NEU2 is involved in various cellular events, including cancer metabolism, neuronal and myoblast differentiation, proliferation, and hypertrophy. However, NEU2-interacting protein(s) within the cell have not been identified yet. Objective The aim of this study is to investigate NEU2 interacting proteins using two-step affinity purification (TAP) strategy combined with mass spectrometry analysis. Methods In this study, NEU2 gene was cloned into the pCTAP expression vector and transiently transfected to COS-7 cells by using PEI. The most efficient expression time of NEU2- tag protein was determined by real-time PCR and Western blot analysis. NEU2-interacting protein(s) were investigated by using TAP strategy combined with two different mass spectrometry experiment; LC-MS/MS and MALDI TOF/TOF. Results Here, mass spectrometry analysis showed four proteins; α-actin, β-actin, calmodulin and histone H1.2 proteins are associated with NEU2. The interactions between NEU2 and actin filaments were verified by Western blot analysis and immunofluorescence analysis. Conclusions Our study suggests that association of NEU2 with actin filaments and other protein(s) could be important for understanding the biological role of NEU2 in mammalian cells.

Background Tyrosinase plays a central role in the biosynthesis pathway of melanin pigment. Melanin protects human skin against radiation and its unusual levels cause some skin disorders such as pregnancy scar, oldness spots and melanoma. Tyrosinase has also been linked to Parkinson’s and other neurodegenerative diseases. In addition, melanin plays a critical role as a defense molecule for insects during wound healing and is important for their life. Therefore, determination of inhibitor molecules for tyrosinase has a promising potential for therapies of some diseases and is an alternative method for keeping insects under control. Material and methods In this study, 1-hepthyl-3-(4-methoxybenzyl)-4 H -1,2,4-triazole-5-one derivative (A6, A8, A15) and 3-(4-chlorophenyl)- 5-(4-methoxybenzyl)-4 H -1,2,4-triazole (B5, B9, B13) derivative compounds were evaluated in terms of their potential for mushroom tyrosinase inhibition. IC 50 values of these six molecules were determined. Results It was seen that B9 molecule was the most effective inhibitor. Docking studies also nearly supported this end result. Tyrosinase inhibition type and K i value were found to be uncompetitive and 370.7±0.3 μM, respectively, in the presence of B9 compound. Conclusion These results suggest that B9 compound is a potential tyrosinase inhibitor.

Background Biotinidase deficiency is a treatable metabolic disease that can be seen with various neurological and dermatological complications. Biomarkers such as alanine, propionylcarnitine (C3) and 3-hydroxyisovalerylcarnitine (C5-OH), which are used to diagnose biotinidase deficiency, are also present. Materials and methods In cases with partial biotinidase deficiency and normal biotinidase activity, alanine, C3 and C5-OH levels were compared in the field by liquid chromatography-tandem mass spectrometry. Results There was no significant difference between subjects with partial biotinidase deficiency and those with normal biotinidase activity between C3 and C5-OH levels. The mean alanine levels in heel blood and plasma were significantly higher than those with normal biotinidase activity in patients with partial biotinidase deficiency. Conclusion In cases with partial biotinidase deficiency, the heel blood alanine level that can be detected in the neonatal screening program may be a leading marker in diagnosis.

Background Gastric cancer, one of the most common cancers in the world, is a multifactorial disease in which environmental and genetic factors play a role. In our study, we aimed to determine the expression levels of four miRNAs (miR127-5p, miR-544a, miR-369-3p and miR-655-3p) on chromosome 14q32 in gastric cancer. Materials and methods Total RNA was isolated from blood samples taken from 66 gastric cancer and 66 healthy individuals. The gene expression levels determined by cDNA and quantitative real-time polymerase chain reaction were analyzed according to the 2 −∆∆Ct method. SPSS 22 were used for statistical analysis and p < 0.05 was considered as statistically significant. Results and discussion miR-655-3p (fold change: 100, p = 0.026), miR-127-5p (fold change: 48, p < 0.001) and miR-369-3p (fold change: 1.6, p > 0.05) was less expressed in the gastric cancer group than control group. miR-544a was found 15.5-fold more expressed in the patient group than control group (fold change: 15.47, p < 0.001). Conclusion miR127-5p, miR-544a, and miR-655-3p may be evaluated as biomarkers in gastric cancer.

Objective Detection of JAK2 V617F in myeloproliferative neoplasms (MPNs) is very important in both diagnosis and disease progression. In our study, we investigated the frequency of JAK2 V617F mutation in patients with myeloproliferative disorders. Methods We retrospectively reviewed the records of 720 patients (174 females and 546 males) who were tested for JAK2 V617F mutation from January 2007 to December 2017. Results In our patients were determined 22.6% JAK2 V617F mutation. 33.3% in women, 19.2% in men have been positive for JAK2 V617F mutation. In our study JAK2 V617F present in 48.6% of essential thrombocythemia, 80.5% of polycythemia rubra vera (PV), 47.5% of primary myelofibrosis, 10% of MPNs, unclassifiable, 0.8% of others. We also investigated the difference in hematological parameters [white blood cell, hemoglobin (Hb), hematocrit (HCT), red blood cell distribution widths (RDW) and platelets count (PLT)] between JAK2 V617F positive and JAK2 V617F negative patients. Conclusions Investigation of the JAK2 V617F mutation is very important in cases of MPNs. In our study JAK2 V617F mutation was higher in PV, essential thrombocythemia, and primary myelofibrosis patients. However, there were significant differences in Hb, HCT, RDW and PLT levels in mutation-positive patients.

Objectives Rituximab is a monoclonal antibody that targets the B-lymphocyte surface antigen CD20. It is used in the treatment of some diseases including B-cell chronic lymphocytic leukemia (B-CLL). There are a lot of data regarding effect of Rituximab on lymphoma cells. But, there is no satisfactory information about the effect of Rituximab on the signaling pathways in leukemia cells. In this study, it was aimed to understand the effect of Rituximab on JAK-STAT and NF-κB signaling pathways in B-cell acute lymphoblastic leukemia (B-ALL) and B-CLL. Material and methods Apoptotic effect of Rituximab in the TANOUE (B-ALL) and EHEB (B-CLL) cell lines were evaluated by using the Annexin V method. mRNA expression levels of STAT3 and RelA were analysed by quantitative RT-PCR (Q-PCR). Alterations in STAT3 and RelA protein expressions were detected by using a chromogenic alkaline phosphatase assay after Western Blotting. Results Rituximab had no apoptotic effect on both cell lines. Complement-mediated cytotoxicity was only detected in EHEB cells. mRNA and protein expressions of STAT3 and RelA genes were decreased following Rituximab treatment. Conclusion Our preliminary results suggest that the use of Rituximab might be effective in B-ALL though both signaling pathways.

Objective DEK is ubiquitously expressed and encodes a nuclear protein, which is also released from some cells. Overexpression of DEK suppresses proliferation of some blood cell progenitors whereas it increases proliferation of epithelial tumors. We showed that DEK is overexpressed in BM cells of 12% of multiple myeloma (MM) patients. Here, we aimed to test if DEK overexpression effects the proliferation and viability of BM stromal cells or MM cells co-cultured with DEK-overexpressing stromal cells, mimicking the BM microenvironment. Methods DEK is stably overexpressed in the BM stromal cell line HS27A. Periodic growth curve and fluorescent activated cell sorting (FACS) analysis was performed to determine the effect of DEK overexpression on HS27A cells and MM cell lines (RPMI-8226 and U266) that are co-cultured with these HS27A cells. Results We showed that, on the contrary to blood progenitors or ephitelial cells, DEK overexpression doesn’t alter the viability or proliferation of the HS27A cells, or the MM cell lines which are co-cultured with DEK-overexpressing HS27A cells. Conclusions Our results suggest that effect of DEK overexpression on the proliferation is cell type and context dependent and increased DEK expression is tolerable by the stromal cells and the co-cultured MM cell lines without effecting proliferation and viability.

Aim The objective of the current study is to examine the association between serum fetuin-A concentrations and some other inflammation markers neutrophil to lymphocyte ratio (NLR), mean platelet volume (MPV) and C reactive protein (CRP) in patients with chronic kidney disease (CKD) and hemodialysis. Methods The study subjects are composed of healthy volunteers (n = 47) and two patient groups; CKD patients (n = 26) and hemodialysis patients (n = 33). We measured serum glucose, urea, creatinine, total protein, albumin, sodium, potassium, calcium, phosphorus, iron, alkaline phosphatase, parathyroid hormone, ferritin and CRP levels by auto-analyzer and fetuin-A levels by ELISA method. Also, complete blood count parameters were analyzed and NLR was calculated. Results There were significant differences in serum fetuin-A concentrations, NLR and MPV values among three groups (p < 0.001, p < 0.001, p < 0.001). The correlation analyses revealed that fetuin-A negatively correlated with urea, creatinine, ferritin, and CRP concentrations (r: 0.349, 0.367, 0.399, 0.550, respectively, p < 0.05). Conclusion Fetuin-A is lower in CKD and hemodialysis patients than the control group; supporting as a negative acute phase reactant. Determination of serum fetuin-A, NLR and MPV might be useful to assess inflammation in CKD and hemodialysis patients.

Background Human serum albumin (HSA) is often selected as a subject of any study because albumin is the most abundant protein in human blood plasma. NMR is recognized as a valuable method to determine the structure of proteins-ligand and protein-drug complexes. Objective – Aim of the study In this study, protein drug interactions were investigated using 5-Fluorouracil anti-cancer drug and human serum albumin protein. Materials and methods In this context 400 MHz NMR spectrometry was used and NMR relaxation rates in drug-albumin complex were investigated with respect to increase albumin concentration and increase in 5-Fluorouracil (5-FU)-albumin solution temperature. Results The results of this study indicated that 5-FU had a weak association with albumin, and it easily dissociated from the protein to which it was attached. Conclusion The obtained results also gave us useful information about molecular dynamics of drug-albumin interactions.

Background Limited research is available concerning the relationship between oxidative stress and inflammation parameters, and simultaneously the effects of rosuvastatin on these markers in patients with hypercholesterolemia. We aimed to investigate the connection between cytokines and oxidative stress markers in patients with hypercholesterolemia before and after rosuvastatin treatment. Methods The study consisted of 30 hypercholesterolemic patients diagnosed with routine laboratory tests and 30 healthy participants. The lipid parameters, interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), paraoxonase-1 (PON1) and malondialdehyde (MDA) levels in controls and patients with hypercholesterolemia before and after 12-week treatment with rosuvastatin (10 mg/kg/day), were analyzed by means of enzyme-linked immunosorbent assay. Results It was found that a 12-week cure with rosuvastatin resulted in substantial reductions in IL-1β, IL-6 and TNF-α and MDA levels as in rising activities of PON1 in patients with hypercholesterolemia. Before treatment, the PON1 levels were significantly negatively correlated with TNF-α and IL-6 in control group, while it was positively correlated with TNF-α in patients. Conclusion Our outcomes provide evidence of protected effect of rosuvastatin for inflammation and oxidative damage. It will be of great interest to determine whether the correlation between PON1 and cytokines has any phenotypic effect on PON1.

Objective The effects of urapidil in intestinal ischemia-reperfusion (IR) model were investigated using histopathological and biochemical methods. Materials and methods Forty Wistar albino rats were subjected to sham operation (Group 1), IR (Group 2), IR+dimethyl sulfoxide (Group 3), IR+urapidil 0.5 mg/kg (Group 4), and IR+urapidil 5 mg/kg (Group 5). Levels of MDA, TAS, TOS, SOD, MPO, NF-κB, caspase-3, and LC3B were measured. Results and discussion The groups 2 and 3 had significantly higher TOS and MPO levels than the sham group had (p < 0.001), whereas the TAS and SOD levels were significantly lower in Group 2 than in the sham group. In treatment groups, TAS and SOD levels increased, whereas TOS, MPO, and MDA levels decreased compared to Group 2. Caspase-3 and LC3B immunopositivities were seen at severe levels in Group 2 and 3. However, Group 4 and 5 were found to have lower levels of immunopositivity. Immunopositivity was observed in interstitial areas, peribronchial region, and bronchial epithelial cells. A moderate level of NF-κB immunopositivity was seen in Group 2 and 3. Conclusion Our results show that urapidil is one of the antioxidant agents and protects lung tissue from oxidant effects of intestinal IR injury.

Background Scavenger receptor class B, type I (SR-BI), involved in reverse cholesterol pathway, is a multilipoprotein receptor and capable of binding HDL, LDL and VLDL. SR-BI may contribute to the development of hypertension due to accumulation of cholesterol in the vessel wall via transporting lipoproteins. Therefore, it was aimed to investigate the relationship between SR-BI rs5888 and rs4238001 variants in the patient with hypertension. Materials and methods Seventy three subjects diagnosed with hypertension and 76 healthy subjects constituted the patient and control group, respectively. Genomic DNA was isolated from peripheral blood samples and a real-time quantitative polymerase chain reaction protocol was performed to detect variations of rs5888 and rs4238001. The results were analyzed with the SPSS 22 program and p < 0.05 was considered statistically significant. Results and discussion SR-BI rs4238001 variation did not show significant difference between patient and control group (p > 0.05). In the SR-BI rs5888 variation; normal homozygous CC and heterozygous CT carriers had an average 2-fold lower risk of hypertension than those carrying the TT genotype (p < 0.05). Conclusion SR-BI rs5888 TT variant may increase hypertension risk by reducing lipid transport to the liver from the vessel wall.

Background Marine-derived fungi are appraised as a favorable source for discovering new bioactive secondary metabolites. In the last few decades researchers have concentrated on marine-derived fungi to obtain new and pharmaceutically active bioactive secondary metabolites with therapeutic potential. Objective In this study three marine-derived fungi were isolated and identified from marine invertebrates and investigated with regard to their antioxidant and cytotoxic activities. Materials and methods DPPH, SO, NO, and ABTS assays were used for monitoring free radical scavenging activity, and the MTT assay was used for testing cytotoxic activity against HCT-116 colon cancer cells. Results According to the obtained results Malassezia restricta extract was shown to have the highest antioxidant and cytotoxic activities compared to the other tested fungi strains. Conclusion This study is the first report about the antioxidant and cytotoxicity activity of Acremonium sclerotigenum , Aspergillus flavus , and M. restricta . This serves as a valuable preliminary study for activity-guided isolation of secondary metabolites.

Aim Our study was aimed to compare two different laboratory methods used to measure serum sodium and serum potassium level i.e. between the indirect Ion Selective Electrode (ISE) based and Enzymatic based method in Beckman Coulter AU-480, USA, in serum samples. Materials and methods This was a cross sectional single center study carried out from September’16 to January’17 on 130 serum samples, to compare the result of serum sodium (S. Na + ) and serum potassium (S. K + ) level by indirect ISE and Enzymatic method in fully automated biochemistry analyzer (Beckman Coulter AU-480, USA). Results To validate new enzymatic method bias, CV% and correlation coefficient were measured for S. Na + and S. K + . The study has drawn a considerable agreement for measurement of S. K + level between two methods with mean ± SD of difference was 0.14 ± 0.22 (p = 0.2242), CV% 2.7% and r 2 value of 0.94, whereas disagreement for measurement of S. Na + level with mean ± SD of difference was 2.38 ± 2.36 (p < 0.01), CV% 0.5% and r 2 value of 0.82. Bias, CV% and correlation coefficient are acceptable for S. K + , whereas nonacceptable for S. Na + level measurement as per Clinical Laboratory Improvement Amendments (CLIA) guidelines. Conclusion There was a good correlation in S. K + between both methods whereas values were not correlated well with S. Na + level. So, enzymatic method can be used as an alternative to indirect ISE method for estimation of S. K + in small laboratories due to its cost effectiveness, but still intense workout is required for estimation of S. Na + .

Background To be able to prevent morbid obesity in the long-term, laparoscopic sleeve gastrectomy (LSG) is one of the most effective surgical interventions. However, leakage and bleeding from the stapler line are significant complications. The aim of this study was to determine the role of the levels of plasma presepsin in the detection of stapler leakage. Materials and methods The study included 300 patients with LSG due to morbid obesity and 40 control subjects. Before any medical treatment was applied, blood samples were taken from patients at 12 h preoperatively and on days 1, 3, and 5 postoperatively. Evaluation was made of plasma presepsin levels, white blood count (WBC), C-reactive protein (CRP) and neutrophil-lymphocyte ratio (NLR), in all patients with sleeve gastrectomy line leakage. Results The WBC, CRP, NLR and presepsin values measured on days 1, 3 and 5 postoperatively were determined to be higher in patients with leakage compared to those without. The predictive value of presepsin (p = 0.001), CRP (p = 0.001) and NLR (p = 0.001) was determined to be statistically significantly higher than that of WBC (p = 0.01). Conclusion The results of the study suggest that presepsin levels could have a role in the detection and follow-up of stapler line leaks after LSG. Elevated presepsin levels, on postoperative day 1 in particular, could have a key role in the early detection of possible complications which are not seen clinically.