Genome-Wide Transcriptional Reorganization Associated with Senescence-to-Immortality Switch during Human Hepatocellular Carcinogenesis

Cataloged from PDF version of article.Senescence is a permanent proliferation arrest in response to cell stress such as DNA damage. It contributes strongly to tissue aging and serves as a major barrier against tumor development. Most tumor cells are believed to bypass the senescence barrier (become "immortal") by inactivating growth control genes such as TP53 and CDKN2A. They also reactivate telomerase reverse transcriptase. Senescence-to-immortality transition is accompanied by major phenotypic and biochemical changes mediated by genome-wide transcriptional modifications. This appears to happen during hepatocellular carcinoma (HCC) development in patients with liver cirrhosis, however, the accompanying transcriptional changes are virtually unknown. We investigated genome-wide transcriptional changes related to the senescence-to-immortality switch during hepatocellular carcinogenesis. Initially, we performed transcriptome analysis of senescent and immortal clones of Huh7 HCC cell line, and identified genes with significant differential expression to establish a senescence-related gene list. Through the analysis of senescence-related gene expression in different liver tissues we showed that cirrhosis and HCC display expression patterns compatible with senescent and immortal phenotypes, respectively; dysplasia being a transitional state. Gene set enrichment analysis revealed that cirrhosis/senescence-associated genes were preferentially expressed in non-tumor tissues, less malignant tumors, and differentiated or senescent cells. In contrast, HCC/immortality genes were up-regulated in tumor tissues, or more malignant tumors and progenitor cells. In HCC tumors and immortal cells genes involved in DNA repair, cell cycle, telomere extension and branched chain amino acid metabolism were up-regulated, whereas genes involved in cell signaling, as well as in drug, lipid, retinoid and glycolytic metabolism were down-regulated. Based on these distinctive gene expression features we developed a 15-gene hepatocellular immortality signature test that discriminated HCC from cirrhosis with high accuracy. Our findings demonstrate that senescence bypass plays a central role in hepatocellular carcinogenesis engendering systematic changes in the transcription of genes regulating DNA repair, proliferation, differentiation and metabolism

Cytotoxic and bioactive properties of different color tulip flowers and degradation kinetic of tulip flower anthocyanins

This study was conducted to determine the potential use of anthocyanin-based extracts (ABEs) of wasted tulip flowers as food/drug colorants. For this aim, wasted tulip flowers were samples and analyzed for their bioactive properties and cytotoxicity. Total phenolic contents of the extracts of the claret red (126.55. mg of gallic acid equivalent (GAE)/g dry extract) and orange-red (113.76. mg GAE/g dry extract) flowers were the higher than those of the other tulip flowers. Total anthocyanin levels of the violet, orange-red, claret red and pink tulip flower extracts were determined as 265.04, 236.49, 839.08 and 404.45. mg pelargonidin 3-glucoside/kg dry extract, respectively and these levels were higher than those of the other flowers. The extracts were more effective for the inhibition of Listeria monocytogenes, Staphylococcus aureus and Yersinia enterocolitica compared to other tested bacteria. Additionally, the cytotoxic effects of five different tulip flower extracts on human breast adenocarcinoma (MCF-7) cell line were investigated. The results showed that the orange red, pink and violet extracts had no cytotoxic activity against MCF-7 cell lines while yellow and claret red extracts appeared to be toxic for the cells. Overall, the extracts of tulip flowers with different colors possess remarkable bioactive and cytotoxic properties. © 2013 Elsevier Ltd

Effects of rootstock and training system on tree canopy, fruit quality and phytochemicals of ‘0900 Ziraat’ and ‘Regina’ sweet cherry cultivars

Both ‘0900 Ziraat’ and ‘Regina’ grafted on ‘Krymsk 5’, or ‘Piku 1’ rootstocks were trained to either Upright Fruiting Offshoots (UFO), Super Slender Axe (SSA) or Kym Green Bush (KGB) training systems. Vegetative growth of the tree, determined by measuring trunk cross-sectional area (TCSA), canopy volume and leaf area, differed significantly, depending on the cultivar x rootstock x training system combination. In general, ‘Krymsk 5’ rootstock resulted in trees with significantly thicker trunks (TCSA: 37.75 cm²) and increased leaf area (up to 86.97 cm²). Fruit weight and fruit quality parameters including Hunter a*, firmness, TSS and acidity were variable between rootstocks and training systems and often not significantly different between treatments. In some years however, significant differences were highly dependent on the training system and rootstock interactions. Higher concentrations of bioactive phytochemical concentrations for total monomeric anthocyanin and antioxidant concentrations were mostly associated with the UFO training system in conjunction with the ‘Krymsk 5’ rootstock suggesting that these are linked to increased tree vigour and increased leaf surface area

Lack of association between the MDM2-SNP309 polymorphism and breast cancer risk

Background: A T-to-G polymorphism (SNP309) at the promoter region of MDM2 has been recently reported to extend the Sp1 binding site that positively regulates the MDM2 transcription level and consequently, its expression level. MDM2 is the negative regulator of p53 tumor suppressor protein and elevated levels of MDM2 hamper the stress response driven by the p53 pathway. Whether MDM2-SNP309 was associated with breast cancer as a predisposing factor was investigated. Patients and Methods: A case-control study of 223 females diagnosed with breast cancer and 149 female controls sampled from the Turkish population was carried out and the T/G MDM2-SNP309 genotype of participants was determined. Results: There was no significant association of the G/G or G/T genotypes with breast cancer risk (odds ratio (OR) 1.14, 95% confidence interval (CI) 0.59-2.22, and OR 1.20, 95% CI 0.67-2.12, respectively). Stratification of the data for onset age or for menopausal status at the time of diagnosis also revealed no association for either group

Transforming growth factor-beta induces senescence in hepatocellular carcinoma cells and inhibits tumor growth

Senescence induction could be used as an effective treatment for hepatocellular carcinoma (HCC). However, major senescence inducers (p53 and p16Ink4a) are frequently inactivated in these cancers.We tested whether transforming growth factor-β (TGF-β) could serve as a potential senescence inducer in HCC. First, we screened for HCC cell lines with intact TGF-β signaling that leads to small mothers against decapentaplegic (Smad)-targeted gene activation. Five cell lines met this condition, and all of them displayed a strong senescence response to TGF-β1 (1-5 ng/mL) treatment. Upon treatment, c-myc was down-regulated, p21Cip1 and p15Ink4b were up-regulated, and cells were arrested at G1. The expression of p16Ink4a was not induced, and the senescence response was independent of p53 status. A short exposure of less than 1 minute was sufficient for a robust senescence response. Forced expression of p21 Cip1 and p15Ink4b recapitulated TGF-β1 effects. Senescence response was associated with reduced nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4) induction and intracellular reactive oxygen species (ROS) accumulation. The treatment of cells with the ROS scavenger N-acetyl-L-cysteine, or silencing of the NOX4 gene, rescued p21Cip1 and p15Ink4b accumulation as well as the growth arrest in response to TGF-β. Human HCC tumors raised in immunodeficient mice also displayed TGF-β1-induced senescence. More importantly, peritumoral injection of TGF-β1 (2 ng) at 4-day intervals reduced tumor growth by more than 75%. In contrast, the deletion of TGF-β receptor 2 abolished in vitro senescence response and greatly accelerated in vivo tumor growth. Conclusion: TGF-β induces p53-independent and p16Ink4a-independent, but Nox4-dependent, p21Cip1-dependent, p15Ink4b-dependent, and ROS-dependent senescence arrest in well-differentiated HCC cells. Moreover, TGF-β-induced senescence in vivo is associated with a strong antitumor response against HCC. Copyright © 2010 by the American Association for the Study of Liver Diseases

Searching for Extraterrestrial Amino Acids in a Contaminated Meteorite: Amino Acid Analyses of the Canakkale L6 Chondrite

Amino acids can serve as important markers of cosmochemistry, as their abundances and isomeric and isotopic compositions have been found to vary predictably with changes in parent body chemistry and alteration processes. Amino acids are also of astrobiological interest because they are essential for life on Earth. Analyses of a range of meteorites, including all groups of carbonaceous chondrites, along with H, R, and LL chondrites, ureilites, and a martian shergottite, have revealed that amino acids of plausible extraterrestrial origin can be formed in and persist after a wide range of parent body conditions. However, amino acid analyses of L6 chondrites to date have not provided evidence for indigenous amino acids. In the present study, we performed amino acid analysis on larger samples of a different L6 chondite, Canakkale, to determine whether or not trace levels of indigenous amino acids could be found. The Canakkale meteor was an observed fall in late July, 1964, near Canakkale, Turkey. The meteorite samples (1.36 and 1.09 g) analyzed in this study were allocated by C. Y. Ornek, along with a soil sample (1.5 g) collected near the Canakkale recovery site

A Convolutional Approach to Vertebrae Detection and Labelling in Whole Spine MRI

We propose a novel convolutional method for the detection and identification of vertebrae in whole spine MRIs. This involves using a learnt vector field to group detected vertebrae corners together into individual vertebral bodies and convolutional image-to-image translation followed by beam search to label vertebral levels in a self-consistent manner. The method can be applied without modification to lumbar, cervical and thoracic-only scans across a range of different MR sequences. The resulting system achieves 98.1% detection rate and 96.5% identification rate on a challenging clinical dataset of whole spine scans and matches or exceeds the performance of previous systems on lumbar-only scans. Finally, we demonstrate the clinical applicability of this method, using it for automated scoliosis detection in both lumbar and whole spine MR scans.Comment: Accepted full paper to Medical Image Computing and Computer Assisted Intervention 2020. 11 pages plus appendi

Phylogeny and S1 Gene Variation of Infectious Bronchitis Virus Detected in Broilers and Layers in Turkey

Citation: Yilmaz, H., Altan, E., Cizmecigil, U. Y., Gurel, A., Ozturk, G. Y., Bamac, O. E., . . . Turan, N. (2016). Phylogeny and S1 Gene Variation of Infectious Bronchitis Virus Detected in Broilers and Layers in Turkey. Avian Diseases, 60(3), 596-602. doi:10.1637/11346-120915-Reg.1The avian coronavirus infectious bronchitis virus (AvCoV-IBV) is recognized as an important global pathogen because new variants are a continuous threat to the poultry industry worldwide. This study investigates the genetic origin and diversity of AvCoV-IBV by analysis of the S1 sequence derived from 49 broiler flocks and 14 layer flocks in different regions of Turkey. AvCoV-IBV RNA was detected in 41 (83.6%) broiler flocks and nine (64.2%) of the layer flocks by TaqMan real-time RT-PCR. In addition, AvCoV-IBV RNA was detected in the tracheas 27/30 (90%), lungs 31/49 (62.2%), caecal tonsils 7/22 (31.8%), and kidneys 4/49 (8.1%) of broiler flocks examined. Pathologic lesions, hemorrhages, and mononuclear infiltrations were predominantly observed in tracheas and to a lesser extent in the lungs and a few in kidneys. A phylogenetic tree based on partial S1 sequences of the detected AvCoV-IBVs (including isolates) revealed that 1) viruses detected in five broiler flocks were similar to the IBV vaccines Ma5, H120, M41; 2) viruses detected in 24 broiler flocks were similar to those previously reported from Turkey and to Israel variant-2 strains; 3) viruses detected in seven layer flocks were different from those found in any of the broiler flocks but similar to viruses previously reported from Iran, India, and China (similar to Israel variant-1 and 4/91 serotypes); and 4) that the AVCoV-IBV, Israeli variant-2 strain, found to be circulating in Turkey appears to be undergoing molecular evolution. In conclusion, genetically different AvCoV-IBV strains, including vaccine-like strains, based on their partial S1 sequence, are circulating in broiler and layer chicken flocks in Turkey and the Israeli variant-2 strain is undergoing evolution. © 2016 American Association of Avian Pathologists

Improvement of breakdown characteristics in AlGaN/GaN/Al xGa 1-xN HEMT based on a grading Al xGa 1-xN buffer layer

To improve the breakdown characteristics of an AlGaN/GaN based high electron mobility transistor (HEMT) for high voltage applications, AlGaN/GaN/Al xGa 1-xN double heterostructure (DH-HEMTs) were designed and fabricated by replacing the semi-insulating GaN buffer with content graded Al xGa 1-xN (x=x 1 → x 2, x 1 > x 2), in turn linearly lowering the Al content x from x 1=90% to x 2=5% toward the front side GaN channel on a high temperature AlN buffer layer. The use of a highly resistive Al xGa 1-xN epilayer suppresses the parasitic conduction in the GaN buffer, and the band edge discontinuity limits the channel electrons spillover, thereby reducing leakage current and drain current collapse. In comparison with the conventional HEMT that use a semi-insulating GaN buffer, the fabricated DH-HEMT device with the same size presents a remarkable enhancement of the breakdown voltage. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Influence of intensive melt shearing on the microstructure and mechanical properties of an Al-Mg alloy with high added impurity content

The official published version can be accessed from the link below - Copyright @ The Minerals, Metals & Materials Society and ASM International 2011We have investigated the influence of melt conditioning by intensive shearing on the mechanical behavior and microstructure of Al-Mg-Mn-Fe-Cu-Si alloy sheet produced from a small book mold ingot with high added impurity content. The melt conditioned ingot has fine grains throughout its cross section, whereas a conventionally cast ingot, without melt shearing, has coarser grains and shows a wider variation of grain size. Both needle-shaped and coarse Chinese script iron bearing intermetallic particles are found in the microstructure at the center of the conventionally processed ingot, but for the melt conditioned ingot, only fine Chinese script intermetallic particles are observed. In addition to the iron bearing intermetallics, Mg2Si particles are also observed. The ingots were rolled to thin sheet and solution heat treated (SHT). During rolling, the iron-based intermetallics and Mg2Si particles are broken and aligned along the rolling direction. Yield strength (YS), ultimate tensile strength (UTS), and elongation of the intensively melt sheared and processed sheet are all improved compared to the conventionally cast and processed sheet. Fractographic analysis of the tensile fracture surfaces shows that the clustered and coarse iron bearing intermetallic particles are responsible for the observed reduction in mechanical properties of the conventionally cast sheet. We have shown that by refining the initial microstructure of the ingot by intensive shear melt conditioning, it is possible to achieve improved mechanical properties at the final sheet gage of an AlMgMn alloy with a high content of impurities.This study is under the Technology Strategy Board funded REALCAR projec