Effects of Complement Regulators and Chemokine Receptors in Type 2 Diabetes.

CD55 and CD59 are complement regulatory proteins suggested to be related with progression of diabetes and its complications. The stromal cell-derived factor 1 (SDF-1) and C-X-C chemokine receptor type 4 (CXCR-4) are chemokine proteins. We aimed to investigate the relation of CD55 and CD59 expression levels and polymorphisms of SDF-1 and CXCR-4 with type 2 diabetes mellitus (T2DM) and its complications. Seventy-five T2DM patients and 73 controls were enrolled. Expression levels of CD55 and CD59 were measured by FACS Calibur; qRT-PCR was used to determine SDF-1 and CXCR-4 gene polymorphisms. CD55 and CD59 expressions in patients with nephropathy, retinopathy and cardiovascular disease were significantly lower than controls. Frequency of CXCR-4 T allele carrying was high in patients and created 1.6 fold risk for the disease (p = .07). CXCR-4 a allele carriers had decreased nephropathy; although there was no statistical significance in carrying CXCR-4 T allele, presence of nephropathy was approximately 2 times higher (p = .254). The nephropathy risk increased 10-fold in CXCR-4 TT genotype carriers (p = .02). All SDF-1 CC genotype carriers had retinopathy, so, it was considered that the CC genotype was effective in retinopathy development (p = .031). For the presence of cardiovascular disease, significant difference was observed for SDF-1 genotypes. Increased cardiovascular risk of 5- and 1.9-fold in SDF-1 T (p = .007) and CXCR-4 T (p = .216) allele carriers, respectively, was observed. We suggest that CD55 and CD59 protein levels and SDF-1 and CXCR-4 have predictive importance in process, complications and tendency of T2DM.

miRNA and mRNA expression profiling in rat brain following alcohol dependence and withdrawal.

Long- lasting alterations in brain gene expression in alcohol addiction have been determined although no clear mechanism has yet been elucidated. There exist many factors regulating the mechanism of gene expression. We aimed in this study to detect miRNA (microRNA) and mRNA expression profile at the specific brain regions regarding ethanol exposure and withdrawal. Rats were exposed to liquid alcohol consumption for 21 days. Oligonucleotides microarrays and bioinformatics analyses were used to identify gene expression, miRNA expression and their functions in the Prefrontal cortex, Hippocampus and Corpus striatum of wistar rats.  A bioinformatics strategy with microarray analysis, quantitative real time PCR, bioinformatics and mRNA (messenger RNA) miRNA- miRNA integrative analyses revealed that expression models interact with neuroplasticity and synaptic processes. Those significantly changed after ethanol exposure and withdrawal processes included 160 mRNAs and 29 rat-miRNAs at prefrontal cortex, 142 mRNAs and 26 rat-miRNAs at hippocampus, and 143 mRNAs and 30 rat-miRNAs at corpus striatum. Gene ontology and ingenuity pathway analyses revealed that most of the altered genes were responsible for synaptic plasticity, neuron differentiation, chromatin organization and some certain important signaling pathways. In conclusion, consistent and integrated variations in miRNA expression and in their focus mRNAs in rat brain were noted after alcohol exposure and withdrawal. Besides, understanding the molecular mechanisms of alcohol abuse will no doubt guide to development of significant cure methods for addiction. We are of the opinion that our findings may shed light on classification of novel biomarkers.

Serum concentrations of Cyclophilin A in patients with Nonalcoholic Fatty Liver Disease.

AIM: Nonalcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver disease and its incidence is rising worldwide. Cyclophilin A (CyPA) is a protein, which is secreted under the presence of oxidative stress and hyperglycemia, and it plays role in proinflammatory signal reduction. In this study we investigated serum levels of CyPA in patients with biopsy proven NAFLD and examined their association with clinical and histological phenotypes. METHODS: In this study, we identified serum levels of CyPA in patients with NAFLD (n=52) and healthy controls without evidence of any liver disease (n=44). The levels of CyPA were measured by enzyme-linked immunosorbent assay and were compared between two study groups. Furthermore, serum levels of CyPA were assessed in relation to the clinical characteristics of the study participants. RESULTS: Serum levels of CyPA were significantly higher in patients with NAFLD (3,8±2,6 µg/ml, P=0.03) compared to healthy controls (2,8±1,8 µg/ml). Moreover, concentrations of CyPA were 2,8±1,8, 3,4±2,3, and 4,2±2,9 µg/ml in control group, non-diabetic and diabetic NAFLD patients, respectively. The difference between the groups was statistically significant (P=0.04). There was significant correlation between the serum concentrations of CyPA and glucose levels (P=0.01), but there was no significant correlation with other clinical and histologic parameters. CONCLUSIONS: Our data suggest that CyPA levels are elevated in patients with NAFLD, especially in patients with diabetes. (Acta gastroenterol. belg., 2017, 80, 3-7).

A novel AVP gene mutation in a Turkish family with neurohypophyseal diabetes insipidus.

PURPOSE: Familial neurohypophyseal diabetes insipidus (FNDI) is a rare, autosomal dominant, inherited disorder which is characterized by severe polydipsia and polyuria generally presenting in early childhood. In the present study, we aimed to analyze the AVP gene in a Turkish family with FNDI. METHODS: Four patients with neurohypophyseal diabetes insipidus and ten healthy members of the family were studied. Diabetes insipidus was diagnosed by the water deprivation test in affected family members. Mutation analysis was performed by sequencing the whole coding region of AVP-NPII gene using DNA isolated from peripheral blood samples. RESULTS: Urine osmolality was low (<300 mOsm/kg) during water deprivation test, and an increase more than 50 % in urine osmolality and recovery of the symptoms were observed by the administration of desmopressin in all patients. Plasma copeptin levels were lower than expected according to plasma osmolality. Pituitary MRI revealed partial empty sella with a bright spot in index patient and a normal neurohypophysis in the other affected subjects. Genetic screening revealed a novel, heterozygous mutation designated as c.-3A>C in all patients. CONCLUSION: c.-3A>C mutation in 5'UTR of AVP gene in this family might lead to the truncation of signal peptide, aggregation of AVP in the cytoplasm instead of targeting in the endoplasmic reticulum, thereby could disrupt AVP secretion without causing neuronal cytotoxicity, which might explain the presence of bright spot. The predicted effect of this mutation should be investigated by further in vitro molecular studies.

Investigation of some DNA repair genes association in non small cell lung cancer.

Ribonucleoside-diphosphate reductase subunit M2, also known as ribonucleotide reductase small subunit, is an enzyme that in humans is encoded by the RRM2 gene and also Ribonucleoside-diphosphate reductase large subunit is an enzyme that in humans is encoded by the RRM1 gene. RRM1 is a gene important in determining tumor phenotype, but also induced the expression of PTEN tumor suppressor gene, cell migration, invasion and metastasis formation, and play a preventive role. ERCC2 DNA repair mechanism is associated in more than 20 genes involved in the NER pathway. The aim of this study is to investigate rs13181 ERCC2 (T>G) (Lys751Gln), rs12806698 RRM1 (-269C>A) and rs6759180 (located in the 5'UTR) RRM2 (10126436G>A) gene polymorphisms by using real time PCR technique in patients with NSCLC. 193 NSCLC cases and 141 healthy control cases were included in this study. A significant difference was found between rs12806698 RRM1 genotype distributions (*p: 0.034) and were determined increases the risk of disease approximately 3.044 times AA genotype having (*p: 0.014 OR: 3.044, 95%CI: 1.205-7,688). A significant difference was found between rs6759180 RRM2 genotype distributions (*p: 0.033) and were determined increases the risk of disease approximately 3.49 times GG genotype having (p: 0,009 OR: 3, 49, %95CI:1.291-9,482). It was found significant difference in serum 8-OHdG levels between patients and controls (*p: 0001).

Determination of effective miRNAs in wound healing in an experimental Rat Model.

The larvae of Lucilia sericata have been used for centuries as medicinal maggots in the healing of wounds. The present study aimed to screen potential microRNAs related to ES-induced wound healing in rat skin wounds and to investigate the potential mechanisms contributing to accelerated wound healing. Healthy, male, 12 weeks old Wistar albino rats weighing 250-300 g were supplied by the Animal Experimental Center. All animal studies were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals. Wistar albino rats were treated by ES after post wounding and the differentially expressed miRNAs in wound biopsies were screened by microarray analysis at the end of treatments for 4,7 and 10 days. In addition, bioinformatics approaches were used to identify the potential target genes of differentially expressed miRNAs and the functions of their target genes. We found a significant up-regulation of rno-miR-99a* and rno-mir-877 in response to ES treatment. Further investigation of rno-miR-99a* and rno-mir-877 and their target genes (TGFa, TNF, TAGLN, MAPK1, MMP-9) implicated in present study could provide new insight for an understanding lead to the development of new treatment strategies. The identified miRNAs can be new biomarkers for ES- induced wound healing.

Investigation of NF-κB1 and NF-κBIA gene polymorphism in non-small cell lung cancer.

Lung cancer is a complex, multifactorial disease which is the leading cause of cancer death in both men and women. NF-κB is a transcription factor which is known to affect the expression of more than 150 genes related to inflammation, lymphocyte activation, cell proliferation, differentiation, and apoptosis, as well as contributing to cell apoptosis and survival. However, NF-κBIA (IκBα) is the inhibitor of the transcription factor. The--94ins/delATTG polymorphism of the NF-κB1 gene promoter region which causes a functional effect and NF-κBIA 3'UTR A → G polymorphism has been shown to be related to various inflammatory diseases and cancer. Ninety-five NSCLC patients and 99 healthy controls were included in study. The NF-κB1-94ins/delATTG and NF-κBIA 3'UTR A → G polymorphism have been studied by using PCR-RFLP method. It was found that the NF-κB1 -94ins/delATTG DD genotype and D allele frequencies were higher in patients than healthy controls and the presence of the DD genotype has a 3.5-fold increased risk of the disease (P: 0.014). This study is the first to investigate the NF-κB1-94ins/delATTG and NF-κBIA 3'UTR A → G polymorphism together in the Turkish population. According to the results, the NF-κB1-94ins/del ATTG promoter polymorphism may have a role in lung carcinogenesis and prognosis.

Identification of a differential expression signature associated with tumorigenesis and metastasis of laryngeal carcinoma.

OBJECTIVES: Metastasis is the most significant prognostic factor for laryngeal carcinoma which necessitates the identification of molecular alterations associated with metastasis. The identification of such molecular alterations will not only prove useful in treatment but also provide insight into mechanisms of cancer metastasis. The studies conducted so far have not specifically focused on metastasis or invasion pathways. Therefore we investigated the expression profiles with a pathway focused approach. MATERIALS AND METHODS: Total RNA was extracted from 36 laryngeal tumors and paired cancer free tissue. Expression levels of 88 genes were determined using a PCR array system following cDNA synthesis. Obtained data was used for the calculation of altered expression levels, facilitating relevant algorithms. Significant alterations were determined according to their p-value obtained by Student's t-test. RESULTS: Sixteen genes have shown altered expression when compared with adjacent cancer-free tissue. 2 of these 16 genes have shown differential expression in tumors with neck metastasis in respect to non-metastatic tumors. CONCLUSION: We found that TGFB1, TIMP1, c-Myc, SPARC, COL4A2 and SOX4 show altered expression in laryngeal tumors. c-Myc and SOX4 expression is decreased as laryngeal tumors switch to metastatic phenotype.

Association between survivin gene promoter -31G/C and -644C/T polymorphisms and non-small cell lung cancer.

Lung cancer is the most common cancer worldwide. Survivin is one of the first reported inhibitors of apoptosis proteins, which is an important family of proteins that regulate apoptosis. The survivin gene is located on human chromosome 17q25, which is composed of 142 amino acids. A common polymorphism of the survivin gene promoter -31G/C has been shown to influence cancer risk. This genetic variant has been associated with overexpression of survivin at both protein and mRNA levels in cancer cells. We examined promoter (-31G/C) genotype frequency in a patient group (N = 146), 77.4% GG, 18.5% GC, 4.1% CC, and in a control group (N = 98), 57.1% GG, 34.7% GC, 8.2% CC. These distributions were significantly different. Promoter (-644C/T) genotype frequency in the patient group was 40.4% TT, 48.6% TC, 11% CC, and in the control group it was 55.1% TT, 40.8% TC, 4.1% CC; these distributions were also significantly different. Individuals carrying the survivin 31 GC genotype and those carrying the survivin 644 CC genotype had a significantly decreased risk of having non-small cell lung cancer.

Circulating resistin is elevated in patients with non-alcoholic fatty liver disease and is associated with steatosis, portal inflammation, insulin resistance and nonalcoholic steatohepatitis scores.

AIM: We aimed to investigate the association between serum levels of resistin and the biochemical and histological features of patients with nonalcoholic fatty liver disease (NAFLD) to determine the usefulness of this relationship in the clinical practice. METHODS: A total of 97 patients with NAFLD and 66 age- and sex-matched healthy controls were recruited. Detailed epidemiological, anthropometric and laboratory data were recorded. Serum levels of resistin were measured with ELISA. RESULTS: Serum levels of resistin were significantly higher in patients with NAFLD (32.10±10.0 ng/mL and 26.57±13.60 ng/mL, respectively) compared with healthy controls (P=0.003). Serum resistin levels were associated with AST, ALT, HOMA-IR, histological steatosis, portal inflammation and nonalcoholic steatohepatitis (NASH) scores. The serum levels of resistin were significantly higher in patients with definite NASH compared to patients with simple steatosis (29±13 and 20±10 ng/mL, respectively, P=0.03). There was no association between the serum resistin levels and the liver fibrosis stages. CONCLUSION: Our data suggest that resistin levels are elevated in patients with NAFLD and could discriminate simple steatosis from definite NASH.