Histone deacetylases in replicative senescence: evidence for a senescence-specific form of HDAC-2

AbstractTo analyze mechanisms of senescence-associated gene expression, we have investigated histone deacetylases (HDACs) in human fibroblasts undergoing replicative senescence. We found that the overall acetylation pattern of histones does not vary detectably with replicative senescence. By Northern blot and Western blot, we found a significant decrease in the abundance of HDAC-1 in senescent cells. Biochemical analysis of deacetylase activities in extracts from old and young cells revealed a striking difference. While by anion exchange chromatography we found a single peak of activity in extracts from young cells, which coincided with the elution of both HDAC-1 and HDAC-2, in senescent cells a second peak of activity was found. This second peak of activity is associated with HDAC-2 but does not contain HDAC-1. These results suggest that HDAC-2 is present in at least two distinct forms, one of which is specific for senescent cells. Further biochemical characterization of the enzyme activity revealed that addition of nicotinamide adenine dinucleotide (NAD) did not detectably influence the activity of any fraction, suggesting that NAD is not an essential co-factor for the analyzed HDACs from diploid human fibroblasts

Origin of positive magnetoresistance in small-amplitude unidirectional lateral superlattices

We report quantitative analysis of positive magnetoresistance (PMR) for unidirectional-lateral-superlattice samples with relatively small periods (a=92-184 nm) and modulation amplitudes (V_0=0.015-0.25 meV). By comparing observed PMR's with ones calculated using experimentally obtained mobilities, quantum mobilities, and V_0's, it is shown that contribution from streaming orbits (SO) accounts for only small fraction of the total PMR. For small V_0, the limiting magnetic field B_e of SO can be identified as an inflection point of the magnetoresistance trace. The major part of PMR is ascribed to drift velocity arising from incompleted cyclotron orbits obstructed by scatterings.Comment: 12 pages, 9 figures, REVTe

Planar cyclotron motion in unidirectional superlattices defined by strong magnetic and electric fields: Traces of classical orbits in the energy spectrum

We compare the quantum and the classical description of the two-dimensional motion of electrons subjected to a perpendicular magnetic field and a one-dimensional lateral superlattice defined by spatially periodic magnetic and electric fields of large amplitudes. We explain in detail the complicated energy spectra, consisting of superimposed branches of strong and of weak dispersion, by the correspondence between the respective eigenstates and the ``channeled'' and ``drifting'' orbits of the classical description.Comment: 11 pages, 11 figures, to appear in Physical Review

JC Virus T-Antigen Regulates Glucose Metabolic Pathways in Brain Tumor Cells

Recent studies have reported the detection of the human neurotropic virus, JCV, in a significant population of brain tumors, including medulloblastomas. Accordingly, expression of the JCV early protein, T-antigen, which has transforming activity in cell culture and in transgenic mice, results in the development of a broad range of tumors of neural crest and glial origin. Evidently, the association of T-antigen with a range of tumor-suppressor proteins, including p53 and pRb, and signaling molecules, such as β-catenin and IRS-1, plays a role in the oncogenic function of JCV T-antigen. We demonstrate that T-antigen expression is suppressed by glucose deprivation in medulloblastoma cells and in glioblastoma xenografts that both endogenously express T-antigen. Mechanistic studies indicate that glucose deprivation-mediated suppression of T-antigen is partly influenced by 5′-activated AMP kinase (AMPK), an important sensor of the AMP/ATP ratio in cells. In addition, glucose deprivation-induced cell cycle arrest in the G1 phase is blocked with AMPK inhibition, which also prevents T-antigen downregulation. Furthermore, T-antigen prevents G1 arrest and sustains cells in the G2 phase during glucose deprivation. On a functional level, T-antigen downregulation is partially dependent on reactive oxygen species (ROS) production during glucose deprivation, and T-antigen prevents ROS induction, loss of ATP production, and cytotoxicity induced by glucose deprivation. Additionally, we have found that T-antigen is downregulated by the glycolytic inhibitor, 2-deoxy-D-glucose (2-DG), and the pentose phosphate inhibitors, 6-aminonicotinamide and oxythiamine, and that T-antigen modulates expression of the glycolytic enzyme, hexokinase 2 (HK2), and the pentose phosphate enzyme, transaldolase-1 (TALDO1), indicating a potential link between T-antigen and metabolic regulation. These studies point to the possible involvement of JCV T-antigen in medulloblastoma proliferation and the metabolic phenotype and may enhance our understanding of the role of viral proteins in glycolytic tumor metabolism, thus providing useful targets for the treatment of virus-induced tumors

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Osteogenic sarcoma (osteosarcoma) in the elderly: Tumor delineation and predisposing conditions

Osteogenic sarcoma (OS), the most common primary bone cancer, is conventionally a primary intramedullary (conventional OS) high-grade malignant tumor characterized by malignant cells forming immature bone or osteoid. The age distribution data for primary bone sarcomas are bimodal. OS is largely a disease of the young but about one-third of OS occurs in patients over 40 years of age. Thus, though considered as rare occurrences, bone tumors occur also in the geriatric population. In this report, tumor delineation and the significance of predisposing conditions to the occurrence of OS are reviewed

PKM2 (pyruvate kinase isoenzyme type M2)

Review on PKM2 (pyruvate kinase isoenzyme type M2), with data on DNA, on the protein encoded, and where the gene is implicated