Anti-cancer efficiency of natural killer cells differentiated from human adipose tissue-derived mesenchymal stem cells and transfected with miRNA150.

AIM: The aim of this study is to investigate the effects of miR150 transfection on NK-like cells differentiated from adipose tissue derived mesenchymal stem cells (AD-MSCs). METHODS: NK-like cells were differentiated from AD-MSCs and activated by miR150 transfection. Transfected/non-transfected NK-like cells were characterized by immunohistochemical and RT-PCR analyzes. Apoptotic efficiency of the transfected/non-transfected NK-like cells on pancreatic cancer cells PANC1 were determined by TUNEL and RT-PCR. RESULTS: In miR150-transfected cells, the increased expression of NK cell-specific genes such as GKMB, KIR2DL2, CD16, CD56, NKG2D, NKp46 and increased immunoreactivity of NK cell-specific surface marker CD314 (NKG2D) were evident. TUNEL assays showed that NK-like cells with/without transfection induced apoptosis in PANC1 cells in the same manner. The decrease in oncogene expression and the increase in the tumor suppressor gene expression in PANC1 cells upon co-culture with NK-like cells differentiated from AD-MSCs were more prominent following miRNA150 transfection. CONCLUSION: It was shown in vitro that NK-like cells could be obtained by differentiation from AD-MSCs and their efficiency could be increased via miR150 transfection. The results are encouraging for further clinical studies in improvement of immunotherapeutic approaches for cancer therapy.

The Effect of Long Term Pre/postnatal Low/high Dose Nicotine Exposure on Tissue Oxidant/antioxidant Status and DNA Damage in Rats.

BACKGROUND: Most women do not stop smoking either during pregnancy or in the lactation period. This study was carried out to investigate the effect of long term per oral pre/postnatal low/high dose nicotine exposure on fetal plasma/tissue oxidant-antioxidant status in rats. METHODS: The study groups were composed of pups whose parents used or did not use nicotine in pregnancy and lactation period. The pups were divided into 3 groups, each consisting of 10 rats; the control group (normal drinking water), low and high dose nicotine groups according to the dose of nicotine (0.4 mg/kg and 6.0 mg/kg BW/day, respectively) given per oral in drinking water. At the end of the 12(th) month, tissue/hemolysate/plasma oxidant-antioxidant status parameters and 8-hydroxy-2-deoxy-guanosine levels were measured. RESULTS: Plasma cotinine levels were higher in nicotine groups compared to controls (p<0.01). A significant increase in liver malonyldialdehyde levels (p<0.05) and a significant decrease in kidney superoxide dismutase activities (p<0.05) were determined in both nicotine groups compared to controls while no statistically significant difference was found in the other parameters. CONCLUSION: This investigation showed that long term nicotine exposure during-after pregnancy may have an adverse effect on vital organs of the offspring via impairing tissue oxidant/antioxidant balance. Liver and kidney seem to be the mostly affected organs possibly due to their major roles in nicotine metabolism.

DNA damage and its relationship with other oxidative stress parameters in Behcet's disease.

Behçet's disease (BD) is a multisystemic, chronic inflammatory, relapsing disorder that is characterized by oral/genital ulcerations, ocular, arthritic, vascular, and neurologic involvements. Recent findings suggest the role of increased oxidative stress and insufficient antioxidant defence system in BD pathogenesis. It has been proposed that the increase in phagocytic cell activity by triggering oxidative reactions in various targets such as lipids, proteins, and DNA leads to severe inflammatory and degenerative pathologies seen in BD In this study, oxidant/antioxidant status of patients with BD was evaluated in comparison with controls and in respect to disease activity by measuring serum nitrite/nitrate, vitamin A, malondialdehyde (MDA), 8-hydroxy deoxyguanosine (8-OHdG), and total sulfhydryl levels (T-SH). The increase in serum MDA and 8-OHdG levels (respectively 30.04 vs. 17.93 nmol/ml, P = 0.0004 and 1.60 vs. 1.03 ng/ml, P = 0.0019) and the decrease in T-SH levels of patients with BD in comparison with controls (0.69 vs. 0.76 mmol/l, P = 0.0085) all indicate the impaired oxidant/antioxidant status in BD. The positive correlation found between MDA/8-OHdG levels (P = 0.02), and the negative correlations both between T-SH/8-OHdG levels (P = 0.031) and T-SH/MDA levels (P = 0.009) show the concordance between the parameters evaluating oxidant-antioxidant status. Among the parameters used for evaluating oxidant/antioxidant status, serum 8-OHdG was the only one showing significantly higher levels in patients with clinically active disease in comparison (P = 0.004) to patients in inactive period. Therefore, 8-OHdG that is assessed for the fist time in BD with this study can be proposed as a more reliable indicator of oxidant stress in evaluating disease activity.

Increased oxidative DNA damage in lean normoglycemic offspring of type 2 diabetic patients.

OBJECTIVE: Several studies have shown increased oxidative stress in patients with pre-diabetes and newly diagnosed Type 2 diabetes mellitus (T2DM). It has been proposed that oxidative stress initiates insulin resistance in genetically predisposed individuals. The aim of this study was to evaluate the markers of oxidative stress in the offspring of patients with T2DM. MATERIAL AND METHODS: We examined 60 lean normoglycemic offspring of Type 2 diabetics, and 52 age, sex and body mass index matched subjects without family history of T2DM as controls. Anthropometric, biochemical and carotid intima media thickness (IMT) measurements and oral glucose tolerance test (OGTT) were performed. Erythrocyte superoxide dismutase and glutathione peroxidase activities, serum nitric oxide, plasma total sulfhydryl (tSH) groups, plasma total antioxidant status, plasma malondialdehyde and serum 8-hydroxydeoxy-guanosine (8-OHdG) levels were compared between 2 groups. RESULTS: 2 groups were similar for the measurements of anthropometric, blood pressure, lipids, fasting glucose, HOMA-IR and carotid IMT. Glucose levels during OGTT were significantly higher in the offspring of Type 2 diabetics than controls (p=0.035). The offspring of Type 2 diabetics showed a significant increase in serum 8-OHdG level (p=0.005) and plasma tSH groups (p=0.032) when compared to the controls. Significant differences were not obtained in other oxidative stress marker levels between 2 groups. CONCLUSION: Main finding of our study was the presence of increased oxidative DNA damage in lean normoglycemic offspring of Type 2 diabetic patients. There is a need for further clinical studies in order to explain whether oxidative stress is present in genetically predisposed subjects and induces the insulin resistance.

Response to growth hormone with respect to pubertal status on increased dose in idiopathic growth hormone deficiency: an analysis of Turkish children in the KIGS database (Pfizer International Growth Study).

AIM: To compare the growth response to growth hormone (GH) treatment in patients with idiopathic GH deficiency (IGHD) who were prepubertal with the response of those who were pubertal at the onset of GH therapy on an increased GH dose. PATIENTS AND METHODS: Among the Turkish patients enrolled in the Pfizer International Growth Study (KIGS) database with the diagnosis of IGHD, the growth data over 2 years of GH therapy were analyzed longitudinally of 113 (79 M) prepubertal (Group 1) and 44 (33 M) pubertal (Group 2) patients. Pubertal signs were reported to be present initially or to have appeared within 6 months of GH therapy in Group 2. Mean +/- SD age at onset of therapy was 8.7 +/- 3.5 and 13.5 +/- 1.8 years; height SDS -4.2 +/- 1.4 and -3.2 +/- 1.1 (p < 0.05) in Groups 1 and 2, respectively. Mid-parental height (MPH) SDS did not show a significant difference between the two groups (-1.5 +/- 1.1 vs -1.7 +/- 1.1). RESULTS: Delta height SDS over 2 years of therapy was significantly higher in Group 1 (1.1 +/- 1.0) than in Group 2 (0.7 +/- 0.6) (p <0.05) in spite of a significantly lower dose of GH (14.6 +/- 3.3 in Group 1 vs 17.0 +/- 3.1 IU/m2/week in Group 2, p < 0.05). Ht--MPH SDS showed an increase from -2.4 +/- 1.7 to -1.4 +/- 1.5 in Group 1 and from -1.5 +/- 1.5 to -0.8 +/- 1.3 in Group 2. Overall delta height SDS showed negative correlations with age (r = -0.32), height SDS (r = -0.41) and height--MPH SDS (r = -0.40) at onset of therapy (p < 0.001). CONCLUSIONS: These data show that in IGHD the slight increase (15-20%) in the dose of GH during puberty was not adequate to maintain height velocity at the same magnitude as in prepuberty, and thus was not cost effective.

A Case with Gaucher-Like Cells.

Gaucher-like cells (GLC) are sometimes indistinguishable from real Gaucher cells. GLC can be detected in various diseases. We present a 4.5 year old boy with massive cervical lympadenopathy and an intraabdominal mass mimicking lymphoma. Many GLC were seen in the fine needle aspiration material of an enlarged lymph node. Ziehl-Neelsen stain of the aspirate revealed many acid-fast bacteria in the GLC. Fine needle aspiration might provide valuable information in the evaluation of enlarged lymph nodes.