The Clinical and Laboratory Efficacy of HA 330 Treatment Combined with Continuous Renal Replacement Therapy in Septic Shock Patients: A Case Series.

INTRODUCTION: Blood purification therapy is a method used to enable cytokine removal and to improve disturbed immune homeostasis in patients with sepsis or septic shock. This study aimed to evaluate the impact of HA 330 treatment on biochemical and hemodynamic parameters and cytokine levels in adult patients with septic shock. METHODS: Critically ill patients with septic shock who received continuous veno-venous hemodiafiltration and HA 330 treatment were included in this prospective observational study. Biochemical and hemodynamic parameters were followed throughout HA 330 treatment. Serum interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor (TNF)-α, high-mobility group box1 (HMGB-1) protein, IL-10 levels were analyzed by ELISA method, before and after each HA 330 session. RESULTS: A total of 18 critically ill patients were included in this study. The median APACHE 2 score was 22.2 ± 7.49 and median SOFA score 9.6 ± 5.44 on intensive care unit admission. SOFA scores were significantly decreased on the 3rd day of HA 330 treatment, compared to 2nd day scores (p = 0.017). Median leukocyte value was significantly decreased (p = 0.027 and p = 0.024), while hemodynamic parameters remained unchanged throughout the HA 330 treatment. Median CRP and procalcitonin levels were significantly reduced at day 3 of HA 330 treatment compared to the baseline (p = 0.015 and p = 0.033, respectively). Serum IL-1 ß, IL-6, IL-8, TNF-a, HMGB-1, and IL-10 levels decreased insignificantly by 11.5%, 26.4%, 11.4%, 37.9%, 0.02%, and 35.5%, respectively, at the end of the hemoperfusion treatment compared to the pre-treatment. CONCLUSION: The administration of HA 330-based hemoperfusion in septic shock patients revealed improvements in SOFA scores, leukocyte count, and CRP and procalcitonin levels. However, there was no statistically significant change in concentrations of inflammatory cytokines and hemodynamic parameters during HA 330 treatment.

SAHA induce hippo pathway in CCA cells without increasing cell proliferation.

BACKGROUND: Cholangiocarcinoma is a malignant tumor originating from bile duct epithelial cells. Since tumor metastasis is associated with poor prognosis and short-term survival of patients, there is an urgent need for alternative therapeutic approaches for CCA. Because of that reason, we aimed to investigate effect of SAHA which is known as HDAC inhibitor on extrahepatic cholangiocarcinoma cell line (TFK-1). METHODS: Cell cycle was measured by Muse Cell Analyzer. YAP, TAZ, TGF-ß protein levels were determined by western-blotting method. TEAD (1-3), TIMP2 and TIMP3 genes level were determined by real-time PCR analysis. RESULTS: We have seen the positive effects of SAHA on the TFK-1 cell line as it reduces cell viability and arresting cells in the G0/G1 phase. We also observed the negative effects of SAHA, as it increases the expression levels of YAP, TAZ, TGF-ß protein and TEAD (1-3) gene. We also found that SAHA reduced the expression levels of TIMP2 and TIMP3 in TFK-1 cells, but was not statistically significant. CONCLUSIONS: Although observing its antiproliferative effects, these negative effects may be related to the cells being resistant to the drug or the remaining cells having a more aggressive phenotype. Therefore, we think that caution should be exercised in the use of this drug for CCA treatment.

Plasma apelin level in patients with restless legs syndrome and its association with periodic leg movements.

OBJECTIVES: Apelin is an antioxidant and anti-inflammatory molecule secreted by adipose tissue and has a protective effect on cardiac and neuronal tissue. Recent studies have reported that the risk of vascular disease is increased in restless legs syndrome (RLS). We aimed to measure plasma levels of apelin in patients with RLS. Additionally, we wanted to determine if there is any relationship between apelin levels and RLS disease severity and the periodic leg movement index (PLMI). METHOD: A total of 14 RLS patients with moderate-to-severe symptoms and 14 age- and body mass index (BMI)-matched healthy controls participated in the study. All participants had no concomitant medical disorder nor took medications. The international RLS rating scale (IRLSS) was used to determine disease severity. Polysomnography (PSG) served to exclude other sleep disorders such as sleep-related breathing disorders and to measure sleep parameters. RESULTS: The mean plasma apelin level was significantly lower in the patient group compared to the control group independent of IRLSS score and PSG findings (p = 0.004). After comparison between the RLS patient group and control group, the patient group was divided into two subgroups based on a PLMI above or below 15 events per hour. A reduced mean apelin level was observed in the patient group having a PLMI above 15 compared to the patient group with PLMI below 15 and the control group (p = 0.003). There was no correlation between plasma apelin levels and disease severity and PLMI in the two patient subgroups. CONCLUSIONS: RLS patients especially those with a PLMI above 15 have low plasma apelin levels independent of disease severity and sleep parameters such as sleep duration and quality. Decreased apelin levels may explain the increased risk for vascular diseases in those patients.

The effects of iodine 131 treatment on chromosomal and oxidative DNA damage in papillary thyroid carcinoma.

Papillary thyroid carcinoma (PTC) is a common endocrine cancer with a good prognosis. Radioactive iodine is thought to be useful for individuals who have had a total or almost total thyroidectomy, but its effects are still controversial. The effects of radioactive iodine-131 (I-131) treatment on oxidative and chromosomal damage in PTC patients were examined in this study, which was carried out with 16 patients newly diagnosed with PTC and 20 healthy control subjects with similar age and gender. Blood samples were taken from patients with PTC at five sampling times (before total thyroidectomy, after total thyroidectomy, and seven days, six months, and one year after treatment) and from control subjects. The cytokinesis block micronucleus cytome (CBMN-cyt) assay parameters in peripheral blood lymphocytes of patients with PTC and controls were evaluated and plasma 8-hydroxydeoxyguanosine (8-OHdG) levels were measured. Furthermore, genome instability and oxidative DNA damage in peripheral blood lymphocytes and plasma of patients with PTC were evaluated before total thyroidectomy (n=16), after total thyroidectomy (before I-131 treatment) (n=16), seven days (n=10), six months (n=5), and one year after treatment (n=5). The numbers of CBMN-cyt assay parameters (micronucleus; MN and nucleoplasmic bridges; NPB) and 8-OHdG levels in patients with PTC were determined to be significantly higher than in those of the control subjects and these values significantly decreased after total thyroidectomy (before I-131 treatment). While the number of MN, apoptotic, and necrotic cells increased after I-131 treatment, it significantly decreased after six months and one year after treatment. The results achieved in this study suggest that I-131 treatment may pose a threat to cells and that radioactive iodine therapy should be avoided (if possible) for patients with PTC after total thyroidectomy.

Serum GAS6, sAXL, IL-10, NO, and BCL-2 levels are decreased in patients with Behçet's disease.

PURPOSE: Behçet's disease (BD) is an autoimmune chronic systemic inflammatory disease characterized by a versatile clinical spectrum. Growth arrest specific protein 6 (GAS6)/soluble AXL (sAXL) signaling pathway draws attention in the resolution of inflammation, and its deficiency is associated with chronic inflammatory, autoimmune diseases, as well as clearance of apoptotic cells by phagocytes - efferocytosis. In this study, it was aimed to investigate whether GAS6/sAXL, interleukin (IL)-10, nitric oxide (NO), and BCL-2 levels were associated with inflammation and efferocytosis contributes to the pathogenesis of BD. METHODS: A total of 37 Behçet patients with ocular involvement and 30 healthy control subjects were included in this study. GAS6, sAXL, IL-10, NO, and BCL-2 levels were quantified using enzyme-linked immunosorbent assay (ELISA) method. RESULTS: Serum GAS6, sAXL, IL-10, NO, and BCL-2 levels were significantly lower in patients with BD compared to the controls (P < 0.005, P < 0.001, P < 0.001, P < 0.001, and P < 0.001, respectively). In correlation analysis, research parameters decreased in patients with BD was significantly correlated with each other: GAS6-IL-10 (r = 0.585, P < 0.001), GAS6-BCL-2 (r = 0.541, P < 0.001), sAXL-BCL-2 (r = 0.696, P < 0.001), IL-10-NO (r = 0.717, P < 0.001), IL-10-BCL-2 (r = 0.759, P < 0.001), and NO-BCL-2 (r = 0.541, P < 0.001). CONCLUSION: In conclusion, decreased serum BCL-2 level may be an indicator of increased apoptosis in these patients and decreased levels of GAS6/sAXL, IL-10, and NO may indicate insufficient clearance of apoptotic bodies released as a result of increased apoptosis in BD patients.

Does dexmedetomidine provide cardioprotection in coronary artery bypass grafting with cardiopulmonary bypass? A pilot study.

OBJECTIVE: The purpose of this pilot study was to evaluate whether dexmedetomidine has a cardioprotective effect during coronary artery bypass graft surgery with cardiopulmonary bypass (CPB). DESIGN: A prospective, double-blind, randomized controlled trial. SETTING: A university hospital. PARTICIPANTS: Thirty-eight patients undergoing coronary artery bypass graft surgery. INTERVENTIONS: Patients were randomized into 2 groups: dexmedetomidine and placebo groups. In the dexmedetomidine group, dexmedetomidine infusion was started by a loading dose of 0.5 µg/kg/10 min, followed by a continuous infusion of 0.5 µg/kg/h. The placebo group received the same volume of saline. Measurements of central venous pressure, mean pulmonary artery pressure (MPAP) and cardiac index were performed before and after dexmedetomidine loading dose and 2, 24 and 48 hours after CPB. Simultaneously, arterial blood was sampled for CK-MB, cardiac troponin T, and N-terminal probrain natriuretic peptide. MEASUREMENTS AND MAIN RESULTS: CK-MB, cardiac troponin T and N-terminal probrain natriuretic peptide values were elevated in the periods after CPB in both groups (p<0.05) and there were no statistically significant differences between groups. MPAP was decreased in the dexmedetomidine group at the 2nd, 24th and 48th hour after CPB (p<0.001, p<0.001, p = 0.002, respectively). Higher cardiac index values were seen earlier in the dexmedetomidine group than in the placebo group (p< 0.05). CONCLUSIONS: Myocardial damage was not reduced by administration of 0.5 µg/kg loading dose and 0.5 µg/kg/h infusion of dexmedetomidine. However MPAP tended to be lower in the dexmedetomidine group. Large-scale clinical outcome studies are indicated to confirm the effect of dexmedetomidine.

Paraoxonase is reduced in patients with growth hormone deficiency: A novel risk factor for atherosclerosis.

BACKGROUND: Growth hormone deficiency (GHD) is associated with an increased cardiovascular mortality. Increased oxidative stress has been associated with development of cardiovascular and cerebrovascular diseases. In the present study, we aimed to evaluate oxidant and antioxidant status in patients with GHD by analyzing serum paraoxonase1 (PON1) activity, and malondialdehyde (MDA) and thiol levels. MATERIALS AND METHODS: This study was a case-control study. Thirty patients with GHD were included in the study and compared with 20 healthy controls. Serum PON1 activity, and MDA and thiol levels were measured according to an enzymatic spectrophotometric method. RESULTS: Serum MDA levels (2.8 ± 1.3 nmol/mL) were higher in GHD group than the controls (1.7 ± 0.5 nmol/mL) (P = 0.001). PON1 activity (149.9 ± 77.9 U/L) was lower in GHD group than the controls (286.3 ± 126.7 U/L) (P = 0.001). Thiol and high-density lipoprotein cholesterol (HDL-cholesterol) levels were lower in GHD group (218.6 ± 103.9 µmol/L and 32.6 ± 13.4 mg/dL, respectively) than the controls (289.6 ± 101.1 µmol/L and 54.3 ± 14.9 mg/dL, respectively) (P = 0.021 and P = 0.001, respectively). In GHD patients, serum MDA level was negatively correlated with serum HDL-cholesterol (r = -0.499, P = 0.001), and serum PON1 activity was positively correlated with serum thiol and HDL-cholesterol levels (r = 0.306, P = 0.032 and r = 0.303, P = 0.033, respectively). CONCLUSION: These data support that GHD is characterized by an imbalance between oxidant and antioxidant factors. This abnormality may contribute to the increased atherogenic risk in patients with GHD.

Gastric ultrasound, citrulline, and intestinal fatty acid-binding protein as markers of gastrointestinal dysfunction in critically ill patients: A pilot prospective cohort study.

BACKGROUND: Gastrointestinal (GI) dysfunction is common in the intensive care unit (ICU), although there is no consensus on biomarkers of GI dysfunction. We aimed to evaluate ultrasound-based gastric antrum measurements and serum intestinal fatty acid-binding protein (IFABP) and citrulline levels in relation to GI dysfunction in critically ill patients. METHODS: Adult critically ill patients receiving enteral nutrition and stayed for in the ICU for ≥48 h was included. GI dysfunction was described using Gastrointestinal Dysfunction Score (GIDS). Gastric antrum measurements, including craniocaudal (CC) diameter, anteroposterior diameter, and antral-cross sectional area (CSA), as well as serum levels for IFABP and citrulline, were prospectively recorded at baseline and on day 3 and day 5 of enteral nutrition. The receiver operating characteristic (ROC) analysis was performed to evaluate gastric ultrasound parameters, serum IFABP, and citrulline concentrations in predicting GI dysfunction. RESULTS: Thirty-nine participants with a median age of 60 years were recruited and 46.2% of participants had GI dysfunction. ROC analysis revealed that the cutoff value of CSA score to predict GI dysfunction was 4.48 cm2 , which provided 72.7% sensitivity and 77.2% specificity (area under the curve = 0.768, 95% CI: 0.555-0.980). At baseline, gastric residual volume was highly correlated with CC diameter and CSA (r = 0.764, P < 0.001 and r = 0.675, P < 0.001, respectively). Serum IFABP and citrulline levels had no correlation with GI dysfunction or gastric ultrasound parameters (P > 0.05). CONCLUSION: CSA was associated with GI dysfunction in critically ill patients. Serum IFABP and citrulline concentrations were poor in predicting GI dysfunction.

Assessment of paraoxonase 1, xanthine oxidase and glutathione peroxidase activities, nitric oxide and thiol levels in women with polycystic ovary syndrome.

OBJECTIVE: To investigate whether there is any relation between oxidative stress and the antioxidant system in the development of polycystic ovary syndrome (PCOS) by measuring serum nitric oxide (NO) levels and xanthine oxidase (XO) activity (a generator of reactive oxygen species) and antioxidant status by measuring serum thiol levels and glutathione peroxidase (GSHPx) and paraoxonase 1 (PON1) activities. DESIGN: Prospective case-control study. SETTING: University hospital in Turkey. SAMPLE: Thirty women with polycystic ovary syndrome and 20 age- and sex-matched healthy control subjects were included. METHODS: Serum XO, PON1 and GSHPx activity and NO and thiol levels were determined by spectrophotometric methods. MAIN OUTCOME MEASURES: Activity of serum XO, PON1 and GSH, as well as NO and thiol levels. RESULTS: Serum XO activities were higher in women with PCOS than in the control women (p<0.001). The PON1 activity was lower in women with PCOS than in the control women (p<0.001). No significant difference was found between NO and thiol levels and GSHPx activities of women with PCOS and the control women (p>0.05). Serum PON1 activities were negatively correlated with serum XO activities and NO levels. CONCLUSION: Increased oxidant XO activity and decreased lipid antioxidant PON1 activity, along with the observed negative correlation between these parameters, suggests that women with PCOS are under oxidative stress and that there is XO-mediated lipid peroxidation, which may be related to increased atherosclerosis seen in later life in such women.

Changes in central corneal thickness, intraocular pressure, and oxidation/antioxidation parameters at high altitude.

BACKGROUND: The authors aimed at investigating ophthalmological changes at high altitude and correlating this with blood oxidation/antioxidation parameters. METHODS: There were 40 volunteers who participated in the study. Initial ophthalmological examinations were performed at 3543 ft (1080 m) and repeated on the following day after the participants climbed to an altitude of 9186 ft (2800 m) on Mt. Erciyes, Turkey. Venous blood samples were taken at both altitudes to evaluate total oxidative system (TOS) and antioxidative system (TAS) levels. RESULTS: IOP-right eyes at 3543 ft (1080 m) was 13.23 +/- 0.43 mmHg and significantly increased to 14.45 +/- 0.56 mmHg at 9186 ft (2800 m). LOP-left eyes at 3543 ft (1080 m) was 13.50 +/- 0.44 mmHg and increased to 14.13 +/- 0.54 mmHg at 9186 ft (2800 m) (P = n.s.). Central corneal thickness (CCT) of the right eyes was 540.98 +/- 4.34 microm at 3543 ft (1080 m) and significantly increased to 549.73 +/- 4.59 microm at 9186 ft (2800 m). CCT of the left eyes was 542.13 +/- 29.01 microm at 3543 ft (1080 m) and significantly increased to 547.23 +/- 4.59 microm at 9186 ft (2800 m). Spherical equivalent refraction of right or left eyes did not show any significant changes. TOS at 3543 ft (1080 m) was 5.33 +/- 0.76 micromol H2O2 equiv/L and significantly increased to 7.55 +/- 0.82 micromol H2O2 equiv/L at 9186 ft (2800 m). TAS at 3543 ft (1080 m) was 2.45 +/- 0.12 micromol H2O2 equiv/L and decreased to 2.22 +/- 0.08 micromol H2O2 equiv/L (P = n.s.) at 9186 ft (2800 m). There was a positive correlation between TAS and LOP at 9186 ft (2800 m). CONCLUSION: Increased CCT can be related to stromal edema caused by hypoxia's effect on corneal endothelial function. Although TOS increased at high altitude, TAS did not show any parallel increase. Since this was nonacclimatized climbing, the antioxidant system could not have reached sufficient levels to counterbalance the observed oxidant stress.

If AFP is elevated, where is cancer? The case report on hereditary persistence of Alpha-fetoprotein.

Alpha-fetoprotein (AFP) is expressed by tumors with a high mitotic index such as hepatocellular carcinoma and germ cell tumors, therefore it is used as a tumor biomarker. Interestingly, although there is no underlying cause, elevated AFP has been reported in some genetically predisposed individuals. This is a very rare and benign condition called "hereditary persistence of AFP (HPAFP)" and an inherited in an autosomal dominant manner. To our knowledge, only 28 families have been reported to date. Some of the reported cases received inappropriate treatments such as chemotherapy and surgery. The possibility of HPAFP should be kept in mind in patients with high AFP in the absence of radiological evidence of hepatocellular carcinoma or germ cell tumor to avoid harmful procedures. It can be easily confirmed by analyzing AFP levels in other family members. We report a case of HPAFP with surprisingly higher AFP levels than previously reported cases and this is the first case reported from Turkey.

Measurement of serial serum total and acylated ghrelin levels in critically ill patients: A prospective and observational pilot study.

BACKGROUND: Ghrelin is a hormone that regulates appetite and energy metabolism. The change of serial serum total and acylated ghrelin levels during hospital stays of critical patients are unknown. In addition, the relationship of this change with the clinical results of patients in the intensive care unit (ICU) is also unknown. The aim of this study was to determine serum total and acylated ghrelin levels serially in critically ill patients. METHODS: This prospective study was performed in the ICU. Patients who were >18 years old and stayed in ICU for >48 h were included in the study. Serum total and acylated ghrelin concentrations were measured at baseline in all participants and serially on the 2nd, 5th, and 10th day after entry into the study in those who remained in the ICU. RESULTS: A total of 60 participants were included. The mean age was 56 ± 21 years. (Baseline, 2nd, 5th, and 10th day median serum total ghrelin levels were 3551 (1651-3995), 3485.20 (1379-4071), 3359 (1167-3919), and 3355 pg/ml (2207-3843), respectively. Baseline, 2nd, 5th, and 10th day acylated ghrelin levels were 47 (0-673), 50 (0-730), 73 (0-808), and 125 pg/ml (0-689), respectively. There was no significant difference between total ghrelin/acylated ghrelin levels and mortality (P > .05). ICU mortality was 30%. CONCLUSION: Ghrelin levels were decreased slightly and acylated ghrelin levels increased substantially over time in critically ill patients. There were no differences between serum total ghrelin/acylated ghrelin levels and ICU mortality .

New Avenues to Explore in SARS-CoV-2 Infection: Both TRIM25 and TRIM56 Positively Correlate with VEGF, GAS6, and sAXL in COVID-19 Patients.

The ongoing COVID-19 pandemic poses a significant threat to human health. Many hypotheses regarding pathogenesis have been proposed and are being tried to be clarified by experimental and clinical studies. This study aimed to reveal the roles of the innate immune system modulator GAS6/sAXL pathway, endothelial dysfunction markers vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF)-1α, and antiviral effective TRIM25 and TRIM56 proteins in pathogenesis of COVID-19. The study included 55 patients with COVID-19 and 25 healthy individuals. The serum levels of GAS6, sAXL, VEGF, HIF-1α, TRIM25, and TRIM56 were measured using commercial ELISA kits and differences between COVID-19 patients and healthy controls, and the relationship to severity and prognosis were evaluated. GAS6, sAXL, TRIM56, and VEGF were found to be higher, while TRIM25 was lower in patients. There were strong positive correlations between GAS6, sAXL, TRIM25, TRIM56, and VEGF. None of the research parameters other than HIF-1α was associated with severity or prognosis. However, HIF-1α was positively correlated with APACHE II. We speculate that the antiviral effective TRIM25 and TRIM56 proteins, as well as the GAS6/sAXL pathway, act together as a defense mechanism in COVID-19. We hope that our study will contribute to further studies to elucidate the molecular mechanism associated with TRIM56, TRIM25, GAS6, sAXL, and VEGF in COVID-19 patients.

Macrolipasemia secondary to colon cancer chemotherapy: a case report.

We reported macrolipasemia in a colon cancer patient during the chemotherapy period without any evidence of pancreatitis. A 52-year-old man formerly treated for papillary thyroid carcinoma had elevated a carcinoembryonic antigen (CEA) concentration in the latest control and was diagnosed with colon cancer. Xelox chemotherapy (oxaliplatin and capecitabine) protocol was planned for six months. Interestingly, the lipase activities gradually increased from 30 U/L to 434 U/L, and exceeded three times the upper limit of the reference range (13-60 U/L). There were no symptoms of pancreatitis, and the abdominal computed tomography (CT) scan was also normal. Polyethylene glycol (PEG) recovery % values of serum samples gradually decreased and were 27% in the recent sample before the end of chemotherapy. Interestingly, the serum lipase activity fell a month after chemotherapy, and PEG recovery % increased (39%). We considered the following possibilities: (1) macrolipasemia due to chemotherapy drugs, (2) macrolipasemia due to antibodies against chemotherapy drugs.

Suberoylanilide Hydroxamic Acid (SAHA) Reduces Fibrosis Markers and Deactivates Human Stellate Cells via the Epithelial-Mesenchymal Transition (EMT).

Hepatic fibrosis is known as the accumulation of connective tissue secondary to chronic damage to the liver. Epithelial-mesenchymal transition (EMT) corresponding increase in liver fibrogenesis was shown with immunohistochemistry and PCR-based studies. Suberoylanilide hydroxamic acid (SAHA), a synthetic compound approved as a histone deacetylase inhibitor (HDAC) by the FDA to treat cutaneous T-cell lymphoma is under investigation for the treatment of lung and renal fibrosis. Experimental modeling for hepatic fibrosis can be constructed with an LX2 cell line isolated from human hepatic stellate cells (HSCs). In this study, we aimed to investigate the modulation of SAHA in the pathogenesis of liver fibrosis by detecting the levels of proteins; (E-cadherin (E-cad), N-cadherin (N-cad), Vimentin (Vim), and genes; E-cad, N-cad, Vim, transforming growth factor-beta (TGF-ß), alpha-smooth muscle actin (α-SMA), type 1 collagen (COL1A1), type 3 collagen (COL3A1)) that play a significant role in EMT with the LX2 cell line. We also evaluated the action of SAHA with cell proliferation, clonogenic, and migration assay. Cell proliferation was performed by flow cytometry. All the protein levels were determined by Western blot analysis, and gene expression levels were measured by Real-Time PCR. Our study observed that SAHA treatment decreased cell viability, colony formation and migration in LX2 cells. We found that SAHA increased E-cad expression level, while it decreased N-cad, Vim, COL1A1, COL3A1, α-SMA TGF-ß genes expression levels. SAHA decreased the level of E-cad, N-cad, and Vim protein levels. We thought that SAHA possesses potent antifibrotic and anti-EMT properties in LX2.

The Effect of EZH2 Inhibition through DZNep on Epithelial-Mesenchymal Transition Mechanism.

Although the molecular pathogenesis of hepatocellular carcinoma (HCC) is uncertain, it is known that the epithelial-mesenchymal transition (EMT) mechanism and epigenetic changes have an important role. This study was focused on evaluating the relationship of 3-Deazaneplanocin A (DZNep) with the EMT mechanism, which is a histone methyltransferase inhibitor on HCC and is also known as an enhancer of zeste homolog 2 (EZH2) inhibitor. Cell viability of HepG2 cells (HCC cell line) assessed for DZNep over 72 hours with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Additionally, colony-forming assay, apoptosis assay, RNA isolation, cDNA synthesis, and real-time PCR (RT-PCR) were performed to see the effect of DZNep on HepG2 cells. DZNep reduced cell proliferation for 72 hours, also significantly reduced colony formation in addition it increased the total apoptosis. DZNep on EZH2, E-cadherin, N-cadherin, and Vimentin (Vim) gene expressions was given different results by either decreasing or increasing the expressions. In this study, we observed a positive effect of DZNep on apoptosis and TIMP3 expression level and decreased colony formation. However, it gave complicated results with the level of gene expression E-cadherin and TIMP2, increase the level of Vim and MMP2 expression. Therefore, we think that further studies are necessary to clarify the role of DZNep.

Chromosomal and oxidative DNA damage in non-functioning pituitary adenomas.

INTRODUCTION: Clinically non-functioning pituitary adenomas (NFPA) are common tumours of the pituitary gland and are mainly considered as benign. The primary aim of this study was to research the effects of NFPA on genome instability in patients with non-functioning pituitary adenoma by using the cytokinesis-block micronucleus cytome (CBMN-cyt) assay and 8-hydroxy- 2'-deoxyguanosine (8-OHdG) assay. The second objective of this study was to assess whether there is a relationship between age, pituitary adenoma diameters, 8-OHDG levels, CBMN site assay parameters, and tumour aggressiveness. MATERIAL AND METHODS: The study was performed on 30 patients who had been diagnosed with NFPA and were admitted to the Department of Endocrinology and Metabolism, and 20 healthy subjects of similar age and sex. RESULTS: Micronucleus (MN), nucleoplasmic bridges (NPBs), nuclear bud (NBUD) frequencies, and apoptotic and necrotic cell frequencies in patients with NFPA were found to be significantly higher than in control subjects, and plasma 8-OHdG levels in patients with NFPA were statistically significantly lower than control subjects in this study. CONCLUSIONS: It is believed that this is the first study to evaluate the aggressiveness of tumour with chromosome/oxidative DNA damage in patients with NFPA. However, further studies are needed in order to understand the cause of NFPA aggression and to evaluate these patients in terms of risk of cancer.

Micronucleus frequency in lymphocytes and 8-hydroxydeoxyguanosine level in plasma of women with polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) has recently been linked with genomic instability and DNA damage. The aim of this study was to test genomic damage in women PCOS, using two different methods for assessing damage in both chromosome and base level. The study was performed on 36 newly diagnosed women with PCOS and 29 healthy women as controls. The micronucleus (MN) analysis used as a biomarker of chromosomal/DNA damage was performed in peripheral lymphocytes by cytokinesis-block method. 8-hydroxydeoxyguanosine (8-OHdG) levels used as a reliable marker of oxidative DNA damage were measured in plasma using an ELISA kit. We found that MN frequencies obtained from lymphocytes of the women with PCOS were significantly higher than those of controls (4.1 +/- 1.0 vs. 2.1 +/- 0.6, P = 0.001), whereas, no differences in 8-OHdG level were found between the patients with PCOS and controls (0.5 +/- 0.3 vs. 0.5 +/- 0.2, P = 0.858). These findings indicate that women with PCOS seem to have increased genomic instability, but do not appear to have oxidative DNA damage despite the increased oxidative stress associated with PCOS.

Assessment of DNA damage and plasma catalase activity in healthy term hyperbilirubinemic infants receiving phototherapy.

Phototherapy (PT) is the most widely used form of treatment for unconjugated hyperbilirubinemia. One possible harmful consequence of PT is of a genetic nature. High levels of bilirubin may lead to oxidative damage in newborns: photochemical reactions may produce toxic photoproducts, probably peroxides. In order to investigate this hypothesis further under in vivo conditions, DNA strand-break frequency was examined by means of the comet assay in peripheral lymphocytes of icteric newborns undergoing PT treatment, and the levels of catalase, an antioxidant enzyme, were determined. We analyzed 20 term non-hemolitic hyperbilirubinemic jaundiced neonates before PT ('before PT' group) and just prior to ending PT ('after PT' group) and compared comet scores of these patients with those of 20 healthy term neonates who all had bilirubin levels in the physiological range. Comet scores (tail length, tail moment and %DNA in tail) of the group 'before phototherapy' were 23.5 +/- 16.3, 7.41 (0.97-40.7), 33.0 +/- 12.1, respectively and scores of after phototherapy group were 3.2 +/- 1.8, 0.29 (0.3-3.2), 10.7 +/- 3.7, respectively. Comet scores of the control group were 3.0 +/- 2.9, 0.25 (0.03-3.22), 10.9 +/- 4.5, respectively. Comet scores and plasma catalase activities in hyperbilirubinemic newborns were significantly higher before phototherapy, compared with the values after phototherapy and in the control groups (p < 0.001). There was no statistical difference between the 'after phototherapy' group and the controls (p > 0.05). These results indicate that high serum bilirubin level has genotoxic effects as is evident from the high rate of DNA strand-breaks in jaundiced newborns. Also PT does not cause an increase in DNA oxidation or induce the genotoxic effects of bilirubin. The counteracting effect of higher catalase activities in hyperbilirubinemic newborns may be responsible for the inactivating toxic and DNA-damaging effects of PT.

Systemic oxidative stress in non-arteritic anterior ischemic optic neuropathy.

PURPOSE: To investigate whether the serum total oxidant status (TOS), total antioxidant status (TAS), advanced oxidation protein products (AOPP), and thiol parameters could play a role in the pathogenesis of non-arteritic anterior ischemic optic neuropathy (NA-AION). METHODS: In this study, 18 newly diagnosed NA-AION patients and 17 healthy subjects (control group) were included. Serum plasma TOS and TAS, AOPP, and thiol parameters were measured by spectrophotometric method and the results were compared. RESULTS: Mean age of the patients and the controls were 60.8 ± 8.4 and 61.9 ± 9.4 years, respectively (p = 0.729). There were no significant differences between the patients and the control group with regard to the values of TAS, TOS, AOPP, and thiol (p = 0.869, p = 0.863, p = 0.040, p = 0.314; respectively). There was a positive correlation between TOS and thiol (p = 0.002, r = 0.681). CONCLUSION: We found no significant relationship between systemic oxidative parameters and NA-AION. However, this study should be accepted as a pilot investigation and needs to be validated.