Orhan Adalı

Background and Objective: Ensuring colon homeostasis is of significant influence on colon cancer and delicate balance is maintained by a healthy human gut microbiota. Probiotics can modulate the diversity of the gut microbiome and prevent colon cancer. Metabolites/byproducts generated by microbial metabolism significantly impact the healthy colonic environment. Hesperidin is a polyphenolic plant compound well known for its anticancer properties. However, low bioavailability of hesperidin after digestion impedes its effectiveness. CYP2W1 is a newly discovered oncofetal gene with an unknown function. CYP2W1 gene expression peaks during embryonic development and is suddenly silenced immediately after birth. Only in the case of some types of cancer, particularly colorectal and hepatocellular carcinomas, this gene is reactivated and its expression is correlated with the severity of the disease. This study aimed to investigate the effects of hesperidin-treated Lacticaseibacillus rhamnosus GG (LGG) cell-free supernatants on CaCo2 colon cancer cell viability and CYP2W1 gene expression. Methods: Alamar Blue cell viability assay was used to investigate the cytotoxic effect of cell-free supernatant of LGG grown in the presence of hesperidin on CaCo2 cells. To observe the effect of cell-free supernatants of LGG on the expression of CYP2W1 gene, qRT-PCR was performed. Results: Five times diluted hesperidin treated cell-free supernatant (CFS) concentration considerably reduced CaCo2 colon cancer cell viability. Furthermore, CYP2W1 gene expression was similarly reduced following CFS treatments and nearly silenced under probiotic bacteria CFS treatment. Conclusion: The CYP2W1 gene expression was strongly reduced by cell-free supernatants derived from LGG culture, with or without hesperidin. This suggests that the suppression may be due to bacterial byproducts rather than hesperidin. Therefore, the CYP2W1 gene in the case of deregulation of these metabolites may cause CYP2W1-related colon cancer cell proliferation.

Objective: Therapeutic properties of Sanguisorba L. genus plants are supported by numerous in vivo and in vitro studies showing the anti-viral, anti-ulcerogenic, anti-cancer, anti-acetylcholinesterase, radioprotective, anti-allergic, and anti-inflammatory activities. The present study was designed to investigate anti-arthritic activity of the “Sanguisorba minor subsp. balearica (Bourg. ex Nyman) Muñoz Garm. & C.Navarro” (Smb), a subspecies of Sanguisorba L. genus, on Complete Freund’s Adjuvant induced (CFA-induced) arthritic rat model. Material and Method: The extract of aerial parts of the Smb was obtained by the consecutive steps of maceration process. The phytochemical content of the extract was analyzed by the High-Performance Liquid Chromatography (HPLC) method. Complete Freund’s Adjuvant-induced arthritic rats were used to examine the anti-arthritic activity of the plant extract. Result and Discussion: HPLC analysis revealed a high amount of phenolic compounds in the Smb extract. The identified phenolics are ellagic acid, gallic acid, quercetin dehydrate and p-coumaric acid with the concentrations of 4.1288; 2.6342; 0.0871 and 0.0633 ppm, respectively. Injection of Smb extract caused a decrease in paw thickness in the CFA-induced arthritic animals. The decrease in paw thicknesses of animals treated by quercetin and Smb 70 mg/kg groups was higher than diclofenac sodium group. Smb 34 mg/kg group also showed a decrease in paw thickness, but it was lower compared to quercetin and Smb 70 mg/kg groups and higher than the diclofenac sodium group. According to the histopathological evaluations of the joint tissues, both 34 mg/kg and 70 mg/kg Smb extract treatment were improved the inflammatory deformations related to arthritis on the rats.

Background Patients with multiple sclerosis (MS) have significantly lower vitamin D levels. Cholesterol is known to be the precursor for vitamin D synthesis, and cholesterol removal is regulated by cholesterol 7α-hydroxylase (CYP7A1) in the liver and cholesterol 24S-hydroxylase (CYP46A1) in the brain. In this study, single nucleotide polymorphisms (SNPs) within the genes CYP7A1 (rs3808607) and CYP46A1 (rs754203) were investigated for their effects on serum lipid profiles, vitamin D levels, and the risk of developing MS. Methods Patients with MS (n = 191) and controls (n = 100) were tested using the PCR–RFLP method to determine their genotypes for rs3808607 and rs754203 SNPs. Results The minor (C) allele frequency for CYP7A1 rs3808607 variation was 0.380 in patients with MS and 0.305 in control subjects (P = .074). For CYP46A1 rs754203, the frequencies of the minor (C) allele were 0.272 and 0.250 in patients and control subjects, respectively (P = .563). Serum vitamin D (25(OH)D3) concentrations were significantly lower in patients than in control subjects (P = .002). The CYP46A1 rs754203 SNP was associated with total cholesterol levels in patients, whereas the CYP7A1 rs3808607 variant was not associated with serum lipid parameters or vitamin D levels in patients or control subjects. Conclusion CYP7A1 rs3808607 and CYP46A1 rs754203 variations are not likely to confer an independent risk for MS development in the Turkish population. To the best of our knowledge, this is the first study to investigate the association between CYP46A1 rs754203 and MS risk.

Purpose: Soluble epoxide hydrolase (sEH) and microsomal epoxide hydrolase (mEH) both catalyze the metabolism of epoxyeicosatrienoic acids (EETs), lipid signaling molecules that are protective against ischemic brain injury owing to their participation in the regulation of vascular tone and cerebral blood flow. In addition, mEH metabolizes polycyclic aromatic hydrocarbons, one of the causative factors of atherosclerotic lesion development. In this study, we aimed to investigate the association of enzyme activity-modifying missense single nucleotide polymorphisms (SNPs) of the sEH gene (EPHX2) and mEH gene (EPHX1) and ischemic stroke risk in a Turkish population. Patients and Methods: Genomic DNA of patients with large artery atherosclerotic ischemic stroke (n=237) and controls (n=120) was isolated from blood samples, and genotypes for Tyr113His (rs1051740) and His139Arg (rs2234922) SNPs of EPHX1 and Arg287Gln (rs751141) SNP of EPHX2 were attained by the PCR/RFLP method. Results: Minor allele frequency and genotype distributions for Arg287Gln, Tyr113His and His139Arg SNPs did not differ significantly between stroke patients and controls. However, hypertension- and diabetes-associated ischemic stroke risk was decreased by EPHX1 and increased by EPHX2 variants in stratification analyses. Conclusion: This study has shown for the first time that the polymorphic alleles of EPHX1 were unlikely to be associated with large artery atherosclerotic ischemic stroke susceptibility; however, protective effects were evident within subgroups of hypertension and diabetes. In addition, EPHX2 Arg287Gln polymorphism, which has been studied for the first time in a Turkish population, was not significantly related to ischemic stroke, but increased the stroke risk in subgroup analysis.

Functional foods are groups of foods that have potentially positive effects on health beyond basic nutrition. Probiotics are beneficial microorganisms, being as a group of functional foods, and when ingested in adequate amounts, they possess positive health effects to host organisms. Lactobacillus acidophilus and Lactobacillus rhamnosus are two of the most studied and very well-known probiotic bacteria. They can have anti-carcinogenic functions, yet their exact mechanisms how they possess anti-carcinogenic activities are still under investigation. Another group of functional foods is plant-based phenolic compounds. They also have potential to possess anti-carcinogenic activities. Among them, oleuropein is very interesting and well-known phenolic compound naturally present in olive fruits and olive oil. As the phenolic compounds and probiotic bacteria can be present at the same time in human gastrointestinal tract, this study aims to investigate the in vitro cytotoxic effects of Lactobacillus acidophilus LA-5 (LA-5) and Lactobacillus rhamnosus GG (GG) grown in the presence of oleuropein on human prostate cancer cell line. To achieve this purpose, oleuropein was added to media of these bacteria and effects of cell-free supernatants of these combinations (phenolic+probiotic) on cytotoxicity of prostate cancer cell line PC-3 were investigated using MTT Method. Concentrations of 0, 50, 100, and 250 µg/mL of oleuropein were added to bacterial cultures and no inhibition on growths of LA-5 or GG were observed. Cell surface hydrophobicity (i.e., adhesion to hydrophobic solvents) is an important feature for bacteria to be retained in gastrointestinal tract (GIT). MATS tests showed all concentrations of oleuropein decreased the surface hydrophobicity of GG, while a dose-dependent increase or decrease in surface hydrophobicity was observed in LA-5. Bacterial aggregation is defined as clustering of bacteria. Auto-aggregation studies showed oleuropein affected the aggregation of bacteria in a dose-dependent manner. Oleuropein added to the growth medium of LA-5 had additional effects on the cytotoxicity of cell-free supernatants of this bacterium. However, addition of oleuropein to the growth medium of GG did not show significant changes on the cytotoxicity of cell-free supernatants of this bacterium. Thus, the results indicate that new combinations of functional foods have potential to formulate new nutraceuticals.

Objectives: Oxidative stress is a known risk factor for the pathogenesis of atherosclerosis, the main cause of ischemic stroke. Glutathione S-transferase (GST) omega-1 and omega-2, members of phase II enzymes, play a role in the antioxidant system. The single nucleotide polymorphisms (SNPs), C419A and A424G in GST omega genes can cause a decrease in enzyme activity. The aim of this study was to investigate the possible association between these polymorphisms and ischemic stroke risk in a Turkish population. Methods: The genotypes and allele frequencies for 239 patients and 130 controls were determined by the PCR/RFLP method. No significant differences were found between patients and controls in terms of genotype and allele frequencies. Results: The frequency of the polymorphic ‘A’ allele was 0.358 in patients and 0.342 in controls for the C419A polymorphism in the GSTO1 gene. The frequency of the polymorphic ‘G’ allele for GSTO2 A424G SNP was 0.370 in patients and 0.404 in controls. The combined homozygous wild type genotype ‘CCAG’ was significantly higher in control group than in the patients. Conclusion: No significant difference was observed between the stroke patients and controls in terms of genotypes and allele distributions. Double combine haplotype CCAA was found to be protective against ischemic stroke when compare to other haplotypes. However, different genotypes of GSTO1 and GSTO2 were observed to have effects on stroke risk in subgroups of diabetics and smokers. In conclusion, the current study is the first to report this finding.

Stroke, a major cause of death and disability, is described as interruption or severe reduction of blood flow in cerebral arteries. Oxidative stress plays an important role in the pathogenesis of atherosclerosis and carotid atherosclerosis is a risk factor for stroke. Combination of multiple environmental and genetic risk factors is thought to increase stroke. Therefore, investigation of the polymorphisms of enzymes is of crucial importance to determine the molecular etiology of the disease. To test this hypothesis, we performed a case–control study in which we compared the distribution of CYP2E1 and NQO1 genotypes between 245 large artery atherosclerotic ischemic stroke patients and 145 controls, using PCR–RFLP. A significant difference was observed between stroke patients and controls with respect to the CYP2E1*5B genotype (odds ratio; OR 8.069, P = 0.011) and allele (OR 7.876, P = 0.011) distribution. However, this polymorphism was not a significant predictor of disease status in logistic regression analysis. NQO1*2 polymorphism genotype distribution was significantly different between patients and controls (P = 0.027) and heterozygote *1*2 genotype was found to be a protective factor against large artery atherosclerotic ischemic stroke in logistic regression analysis (OR 0.562, P = 0.018). This is the first study conducted regarding the association of CYP2E1 and NQO1 genetic polymorphisms and large artery atherosclerotic ischemic stroke risk in Turkish population.

In the present study, the possible role of ellagic acid (EA) on antioxidant potential of Epilobium hirsutum (EH) in rat liver was investigated. Wistar rats were intraperitoneally treated with 37.5 mg/kg of EH and 10 mg/kg of EA for 9 days. Effects of EH and EA on antioxidant [glutathione peroxidase (GPx) and superoxide dismutases (SOD)] and Phase II [NADPH quinone oxidoreductase 1 (NQO1) and glutathione S-transferases (GSTs)] enzyme activities, as well as protein and mRNA expressions of those, were investigated. Polyphenolic content of EH was determined by LC-MS/MS analysis. EH and EA injection to rats resulted in a significant increase of NQO1 (3.6-fold and 4.7-fold), GPx (1.45-fold), and SOD (1.34-fold and 1.27-fold) enzyme activities, whereas total GST (46% and 57%) and its isoforms,and GST mu (57% and 72%), and GST theta (60% and 68%) activities were significantly decreased. Western-blot and qRT-PCR analysis showed that NQO1 and GPx protein and mRNA expressions were increased significantly (P < 0.0001), whereas GST mu and GST theta were significantly decreased (P < 0.0001).