Investigation of microRNA expression changes in HepG2 cell line in presence of URG4/URGCP and in absence of URG4/URGCP suppressed by RNA interference

Hepatocellular carcinoma (HCC) originates from liver cells and is one of the most common malignant cancers in the world. microRNAs (miRNA), are single strand non-coding RNA molecules with the length of 18–25 nucleotides. miRNAs play an important role in the development of HCC, i.e., miRNAs have a significant impact on multistep hepatocellular carcinogenesis including cellular migration and invasion. URG4/URGCP (up-regulated gene-4/upregulator of cell proliferation) is up-regulated in the presence of HBxAg and has been identified and characterized by Satiroglu-Tufan et al. The full-length URG4/URGCP is 3.607?kb. Overexpression of URG4/URGCP in the presence of HBV X protein may function as a putative oncogene that significantly contributes to multi-step hepatocarcinogenesis. In this study, we aimed to investigate potential miRNA expression changes in HepG2 cell line model system in the presence of URG4/URGCP and in the absence of URG4/URGCP, which was suppressed by RNA interference. To functionally characterize URG4/URGCP, independent cultures of HepG2 cells were stably transfected with pcDNA3 or pcDNA3-URG4/URGCP. Relative quantification of whole genome miRNAs was analyzed by RT-PCR using human whole genome miRNA qPCR profiling kits. Among the 1,034 human miRNAs investigated by the arrays, 77 miRNAs were up-regulated and nine miRNAs were down-regulated in the presence of URG4/URGCP. In conclusion, we have analyzed miRNA profiles in HepG2 cells in presence or absence of URG4/URGCP gene using RNA interference. Some of these miRNAs may play roles in URG4/URGCP gene related disease development through the regulation of different signaling pathways.     

Antioxidant Enzyme and Element Status in Heroin Addiction or Heroin Withdrawal in Rats: Effect of Melatonin and Vitamin E Plus Se

Heroin use, withdrawal syndrome, and heroin-related deaths are still the most serious public health problems. Antioxidants and bio-elements are essential for metabolism in living organisms. To our knowledge, there are no data about the effect of antioxidant therapy on the levels of bio-elements and antioxidant enzymes in the naloxone (NX)-induced heroin withdrawal syndrome. Therefore, in the present study for the first time, we have investigated the role of antioxidant therapy, melatonin, and vitamin E plus Se, on the trace and major elements and antioxidant enzymes in the heroin addiction or heroin withdrawal in rats. Glutathione peroxidase levels were increased and catalase levels were decreased in the all study groups when compared to the sham group. The level of superoxide dismutase (SOD) in the fixed dose of heroin (FDH) given group was lower; however, in the variable doses of heroin (VDH) given group SOD level was higher. Furthermore, in withdrawal syndrome, Fe, Mg, Mn, and Ti levels were diminished and Al, Ca, and Cu levels were increased in the FDH+NX group. Moreover, Mg, Mn, and Se levels were also diminished and Al level was increased in the VDH+NX group. In conclusion, our results obviously indicated that heroin effected both bio-element status and antioxidant enzyme activities and, exogenous melatonin or vE+Se therapy might relieve on the element and antioxidant enzyme the destructive activity caused by heroin.     

In vitro genotoxicity assessment of the synthetic plant growth regulator, 1-naphthaleneacetamide

1-Naphthaleneacetamide (NAAm) is a synthetic plant growth regulator in the auxin family that is widely used in agriculture to promote the growth of numerous fruits, for root cuttings and as a fruit thinning agent. The potential genotoxic effects of NAAm were investigated in vitro by the chromosome aberrations (CAs), and cytokinesis-block micronucleus assays in human peripheral blood lymphocytes (PBLs) for the first time. The human PBLs were treated with 20, 40, 80, and 160 µg/mL of NAAm for 24 and 48 h. The results of this study showed that NAAm significantly induced the formation of structural CA and MN for all concentrations (20, 40, 80 and 160 µg/mL) and treatment periods (24 and 48 h) when compared with the negative and the solvent control. In addition, the higher concentrations of NAAm (80 and 160 µg/mL) caused a statistically significant increase in nuclear bud (NBUD) formation for both 24 and 48 h treatment times. With regard to the cell cycle kinetics, at all the tested concentrations, NAAm caused a statistically significant reduction in the mitotic index (MI) only for 48 h treatment period and also in the nuclear division index (NDI) for both 24 and 48 h treatment periods as compared to the control groups. The reductions in the MI and NDI occured in a concentration-dependent manner for both treatment times. In conclusion, the present results indicate that in the tested experimental conditions, NAAm was genotoxic and cytotoxic on human PBLs in vitro.     

Investigation of Antimicrobial,Anti-Quorum Sensing,and Cytotoxic Activities of Flavonoids Isolated from Pulicaria armena Boiss. & Kotschy ex Boiss. (Asteraceae)

This is the first report on the separation and biological assessment of all metabolites derived from Pulicaria armena (Asteraceae) which is an endemic species narrowly distributed in the eastern part of Turkey. The phytochemical analysis of P. armena resulted in the identification of one simple phenolic glucoside together with eight flavon and flavonol derivatives whose chemical structures were elucidated by NMR experiments and by the comparison of the spectral data with the relevant literature. The screening of all molecules for their antimicrobial, anti-quorum sensing, and cytotoxic activities revealed the biological potential of some of the isolated compounds. Additionally, quorum sensing inhibitory activity of quercetagetin 5,7,3’ trimethyl ether was supported by molecular docking studies in the active site of LasR which is the primary regulator of this cell-to-cell communication system in bacteria. Lastly, the critical molecular properties indicating drug-likeness of the compounds isolated from P. armena were predicted. As microbial infections can be a serious problem for cancer patients with compromised immune systems, this comprehensive phytochemical research on P. armena with its anti-quorum sensing and cytotoxic compounds can provide a new approach to the treatment.     

Biosynthesis of isoprenoids, polyunsaturated fatty acids and flavonoids in Saccharomyces cerevisiae

Industrial biotechnology employs the controlled use of microorganisms for the production of synthetic chemicals or simple biomass that can further be used in a diverse array of applications that span the pharmaceutical, chemical and nutraceutical industries. Recent advances in metagenomics and in the incorporation of entire biosynthetic pathways into Saccharomyces cerevisiae have greatly expanded both the fitness and the repertoire of biochemicals that can be synthesized from this popular microorganism. Further, the availability of the S. cerevisiae entire genome sequence allows the application of systems biology approaches for improving its enormous biosynthetic potential. In this review, we will describe some of the efforts on using S. cerevisiae as a cell factory for the biosynthesis of high-value natural products that belong to the families of isoprenoids, flavonoids and long chain polyunsaturated fatty acids. As natural products are increasingly becoming the center of attention of the pharmaceutical and nutraceutical industries, the use of S. cerevisiae for their production is only expected to expand in the future, further allowing the biosynthesis of novel molecular structures with unique properties.     

Theoretical modeling and experimental studies on N-n-Decyl-2-oxo-5-nitro-1-benzylidene-methylamine

The Schiff base compound, N-n-Decyl-2-oxo-5-nitro-1-benzylidene-methylamine, has been -synthesized and characterized by IR, electronic spectroscopy, and X-ray single-crystal determination. Molecular geometry from X-ray experiment of the title compound in the ground state have been compared using the Hartree-Fock (HF) and density functional method (B3LYP) with 6-31G(d) basis set. Calculated results show that density functional theory (DFT) at B3LYP/6-31G(d) level can well reproduce the structure of the title compound. To investigate the solvent effect for the atomic charge distributions of the title compound, self-consistent reaction field theory with Onsager reaction field model was used. In addition, DFT calculations of the title compound, molecular electrostatic potential and thermodynamic properties were performed at B3LYP/6-31G(d) level of theory.