Influenza infection induces host DNA damage and dynamic DNA damage responses during tissue regeneration

Influenza viruses account for significant morbidity worldwide. Inflammatory responses, including excessive generation of reactive oxygen and nitrogen species (RONS), mediate lung injury in severe influenza infections. However, the molecular basis of inflammation-induced lung damage is not fully understood. Here, we studied influenza H1N1 infected cells in vitro, as well as H1N1 infected mice, and we monitored molecular and cellular responses over the course of 2 weeks in vivo. We show that influenza induces DNA damage to both, when cells are directly exposed to virus in vitro (measured using the comet assay) and also when cells are exposed to virus in vivo (estimated via γH2AX foci). We show that DNA damage, as well as responses to DNA damage persist in vivo until long after virus has been cleared, at times when there are inflammation associated RONS (measured by xanthine oxidase activity and oxidative products). The frequency of lung epithelial and immune cells with increased γH2AX foci is elevated in vivo, especially for dividing cells (Ki-67-positive) exposed to oxidative stress during tissue regeneration. Additionally, we observed a significant increase in apoptotic cells as well as increased levels of DNA double strand break (DSB) repair proteins Ku70, Ku86 and Rad51 during the regenerative phase. In conclusion, results show that influenza induces DNA damage both in vitro and in vivo, and that DNA damage responses are activated, raising the possibility that DNA repair capacity may be a determining factor for tissue recovery and disease outcome.     

High nuclear DNA content in nervus terminalis ganglion cells ofScorpaena porcus

Zusammenfassung Gewisse Ganglien-Zellen vonScorpaena porcus weisen einen DNA-Gehalt auf, der 32 bis 64 mal höher ist als derjenige von normalen Neuronen.

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Research supported by a CNR grant.     

Cation inhibition of DNA synthesis in mammary epithelial cells in vitro

Zusammenfassung Lithium- und Ammoniumionen verhindern den Anfang der DNA-Synthese im Milchdrüsenepithel in vitro. Die Replikation von DNA, nach dem Anfang derS-Periode, wird jedoch nicht verhindert. Diese spezifische Verhinderung der Dauer derG ₁-Periode durch die ionale Umgebung zeigt, dass Mechanismen, die für die Regulierung der Zellproduktion wichtig sind, während derG ₁-Periode intervenieren.

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This work was supported in part by grant No. CA 10268 from the U.S. Public Health Service.     

Reaction step requiring protein synthesis in DNA synthesis in sea urchin embryos

Summary In the acid-soluble fraction extracted from embryos treated with cycloheximide, thymidine triphosphate was largely accumulated and DNA polymerase activity decreased, while both activities of thymidine kinase and thymidylate kinase remained unchanged.     

Peripheral inhibition in sustained and transient on-center ganglion cells in cat retina

Zusammenfassung Die Resultate deuten darauf hin, dass nicht, wie bisher vermutet wurde, die Stützzellen schwach auf Diffuslicht reagieren, sondern dass unter bestimmten Umständen die Inhibition stärker bzw. schwächer sein kann.

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This research was supported by Public Health Research Grant Nos. EY00376 and EY00701.     

The number of neurons,glial and perineurium cells in an insect ganglion

Zusammenfassung Im zweiten Abdominalganglion der Indischen Stabheuschrecke (Carausius morosus Br.) wurden 600 ± 50 Neurone, 1000 ± 100 Gliazellen und 500 ± 30 Perineuriumzellen, zusammen etwa 2100 Zellen gezählt. Die Volumina des Ganglions, des Neuropilems und des Zellcortex betrugen etwa: 140 · 10^–4 mm³, 75 · 10^–4 mm³ und 65 · 10^–4 mm³.     

Inhibition by carbaryl of DNA,RNA and protein synthesis in cultured rat lung cells

Summary The carbamate pesticide carbaryl rapidly inhibited DNA, RNA and protein synthesis in L-2 cells from rat lung. the inhibiton was partly reversible and was not accompanied by inhibiton of transport of tritiated precursors into intracellular pools or destruction of the integrity of the cell membrane.We thank W. H. J. Douglas for his gift of L-2 cells.This work was supported by contract 22140 of the Kentucky Tobacco Research Board.     

Nucleolar hypertrophy and nuclear DNA replication in liver

Summary Hypertrophy of nucleoli is associated with DNA synthesis in the hepatocytes of untreated rats just as it is in stimulated animals. Nucleolar enlargement is not a sufficient change to ensure that the parenchymal liver cell can make DNA.This work was supported by a grant from the National Cancer Institute.     

Transport and diffusion of Tau protein in neurons

In highly polarized and elongated cells such as neurons, Tau protein must enter and move down the axon to fulfill its biological task of stabilizing axonal microtubules. Therefore, cellular systems for distributing Tau molecules are needed. This review discusses different mechanisms that have been proposed to contribute to the dispersion of Tau molecules in neurons. They include (1) directed transport along microtubules as cargo of tubulin complexes and/or motor proteins, (2) diffusion, either through the cytosolic space or along microtubules, and (3) mRNA-based mechanisms such as transport of Tau mRNA into axons and local translation. Diffusion along the microtubule lattice or through the cytosol appear to be the major mechanisms for axonal distribution of Tau protein in the short-to-intermediate range over distances of up to a millimetre. The high diffusion coefficients ensure that Tau can distribute evenly throughout the axonal volume as well as along microtubules. Motor protein-dependent transport of Tau dominates over longer distances and time scales. At low near-physiological levels, Tau is co-transported along with short microtubules from cell bodies into axons by cytoplasmic dynein and kinesin family members at rates of slow axonal transport.     

Ethanol and liver protein synthesis in vivo

Summary Ethanol was administered i.p. to adult roosters during hormonally induced vitellogenin synthesis. At moderate doses, ethanol had no influence on the synthesis of vitellogenin nor did it cause alterations in the size distribution of liver polyribosomes.This was supported in part by a grant from the Foundation for Alcohol Studies, Finland.     

Expanding the roles of chromatin insulators in nuclear architecture,chromatin organization and genome function

Of the numerous classes of elements involved in modulating eukaryotic chromosome structure and function, chromatin insulators arguably remain the most poorly understood in their contribution to these processes in vivo. Indeed, our view of chromatin insulators has evolved dramatically since their chromatin boundary and enhancer blocking properties were elucidated roughly a quarter of a century ago as a result of recent genome-wide, high-throughput methods better suited to probing the role of these elements in their native genomic contexts. The overall theme that has emerged from these studies is that chromatin insulators function as general facilitators of higher-order chromatin loop structures that exert both physical and functional constraints on the genome. In this review, we summarize the result of recent work that supports this idea as well as a number of other studies linking these elements to a diverse array of nuclear processes, suggesting that chromatin insulators exert master control over genome organization and behavior.     

Checkpoint kinase 1 in DNA damage response and cell cycle regulation

Originally identified as a mediator of DNA damage response (DDR), checkpoint kinase 1 (Chk1) has a broader role in checkpoint activation in DDR and normal cell cycle regulation. Chk1 activation involves phosphorylation at conserved sites. However, recent work has identified a splice variant of Chk1, which may regulate Chk1 in both DDR and normal cell cycle via molecular interaction. Upon activation, Chk1 phosphorylates a variety of substrate proteins, resulting in the activation of DNA damage checkpoints, cell cycle arrest, DNA repair, and/or cell death. Chk1 and its related signaling may be an effective therapeutic target in diseases such as cancer.     

Differential sensitivity to ethidium bromide of replicative DNA synthesis and bleomycin-induced unscheduled DNA synthesis in permeable mouse sarcoma cells

Replicative DNA synthesis in permeable mouse sarcoma cells was more sensitive to ethidium bromide (EtBr) than bleomycin-induced unscheduled DNA synthesis (UDS). A similar difference in sensitivity to EtBr was observed between DNA polymerases alpha and beta. The difference in sensitivity to EtBr of replicative DNA synthesis and UDS in the present system seems to reflect mainly the sensitivity difference between DNA polymerases alpha and beta.     

Differential sensitivity to ethidium bromide of replicative DNA synthesis and bleomycin-induced unscheduled DNA synthesis in permeable mouse sarcoma cells1

Summary Replicative DNA synthesis in permeable mouse sarcoma cells was more sensitive to ethidium bromide (EtBr) than bleomycin-induced unscheduled DNA synthesis (UDS). A similar difference in sensitivity to EtBr was observed between DNA polymerases and . The difference in sensitivity to EtBr of replicative DNA synthesis and UDS in the present system seems to reflect mainly the sensitivity difference between DNA polymerases and .Acknowledgments. The authors wish to thank Nippon Kayaku Co. (Tokyo, Japan) for providing copper-free bleomycin A₂. This research was supported in part by a grant from the Japan Ministry of Education, Science and Culture.