Fragments of linear unintegrated Rous sarcoma virus DNA, resulting from digestion with SmaI restriction endonuclease]

Linear unintegrated DNA of Schmidt-Ruppin Rous sarcoma virus, subgroup D (SR-RSV-D), was digested with SmaI restriction endonuclease, analyzed by agarose gel electrophoresis, "blotted" by Southern's method and hybridized with a viral 32P cDNA. SmaI cleaves this DNA at five sites, two of which are localized at the ends of the provirus. Src and env genes seem not to be restrictied by SmaI cleavage.     

An increase in Sendai virus-induced cell fusion of erythrocytes infected withPlasmodium chabaudi

Summary The extent of cell fusion induced by Sendai virus was examined in erythrocytes infected withPlasmodium chabaudi. An increase in cell fusion of erythrocytes with Ehrlich tumor cells and of erythrocytes with erythrocytes was observed wit the infected erythrocytes. However, agglutination by the virus was not changed between erythrocytes of normal and malarial mice. These results indicate that the increase in cell fusion occurred in the process of membrane fusion, suggesting that some membrane property ofPlasmodium-parasitized erythrocytes is changed in terms of Sendai virus-induced cell fusion.We thank Drs George L. Gerton, T. Matsuyama and M. Niwa for their comments on this work and Mr I. Kimata for preparing photographs.     

An increase in Sendai virus-induced cell fusion of erythrocytes infected with Plasmodium chabaudi

The extent of cell fusion induced by Sendai virus was examined in erythrocytes infected with Plasmodium chabaudi. An increase in cell fusion of erythrocytes with Ehrlich tumor cells and of erythrocytes with erythrocytes was observed with the infected erythrocytes. However, agglutination by the virus was not changed between erythrocytes of normal and malarial mice. These results indicate that the increase in cell fusion occurred in the process of membrane fusion, suggesting that some membrane property of Plasmodium-parasitized erythrocytes is changed in terms of Sendai virus-induced cell fusion.     

Novel AIDS therapies based on gene editing

HIV/AIDS remains a major public health issue. In 2014, it was estimated that 36.9 million people are living with HIV worldwide, including 2.6 million children. Since the advent of combination antiretroviral therapy (cART), in the 1990s, treatment has been so successful that in many parts of the world, HIV has become a chronic condition in which progression to AIDS has become increasingly rare. However, while people with HIV can expect to live a normal life span with cART, lifelong medication is required and cardiovascular, renal, liver, and neurologic diseases are still possible, which continues to prompt research for a cure for HIV. Infected reservoir cells, such as CD4+?T cells and myeloid cells, allow persistence of HIV as an integrated DNA provirus and serve as a potential source for the re-emergence of virus. Attempts to eradicate HIV from these cells have focused mainly on the so-called “shock and kill” approach, where cellular reactivation is induced so as to trigger the purging of virus-producing cells by cytolysis or immune attack. This approach has several limitations and its usefulness in clinical applications remains to be assessed. Recent advances in gene-editing technology have allowed the use of this approach for inactivating integrated proviral DNA in the genome of latently infected cells or knocking out HIV receptors. Here, we review this strategy and its potential to eliminate the latent HIV reservoir resulting in a sterile cure of AIDS.     

Interferon production by sendai virus-treated hamster and monkey cells

Résumé On présente une étude comparée sur la production d'interféron par les cellules non-transformées et tumorales de hamster ainsi que par les cellules de singe, après leur traîtement par le virus Sendai inactivé. Les cellules tumorales ne produisent pas l'interféron sous l'action du virus sendai. Les cellules non-transformées cessent de produire ou de libérer l'interféron, dans leur milieu de culture, en moins de 24 h après l'enlèvement du virus Sendai de ce milieu.

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Supported by a Fellowship from The Medical Research Council of Canada. We thank Mr.L. Guerin for supplying some of the culture material used, Dr.E. Perry for his support and discussions, and Dr.J. C. N. Westwood for his support. Our present address is: Department of Tumor Biology, Karolinska Institutet, 104 01 Stockholm 60 (Sweden).     

Human spleen: a new and potent source of human interferon for experimental and therapeutic use

Cells of a single human spleen, cultivated in vitro and induced by Sendai virus, produce constantly more than 10(8) units of interferon per spleen. This interferon can be used for therapeutic assays and new basic investigations.     

Cellular fusion induced in vitro by the mouse scrapie agent]

A method is described to demonstrate and measure the cell-fusion in vitro induced by viruses. This technique has been established using Sendai virus. It has been used to study the fusing ability of the Scrapie agent which is responsible of a slowly progressing spongiform encephalopathy in Mice. In Vero cells, the Scrapie agent induces a fusion which appears slowly and remains moderate. This test can help to detect the human spongiform encephalopathies.     

Induction of active immune state by multinucleate tumour cells in mice

Résumé Une immunité active au sarcome, produit par le cholanthrène de méthyle, a été obtenue en utilisant des cellules tumorales multinucléées, associées au virus de Sendai; les réactions ont été spécifiques à la tumeur. A la suite de ces injections, plus de 70% des souris furent résistantes aux trois inoculations, avec 10⁵ cellules vivantes tumorales.     

Cardiotonic activities of some new type of bufadienolide- and cardenolide-conjugates

Summary Some new type of bufadienolide-and cardenolide-conjugates, including bufotoxins and 3-sulfates recently isolated from the toad skin, were tested for their cardiotonic activities by using isolated frog hearts and guinea-pig atria. The relative potencies were obtained and reported.This study was reported at the 49th General Meeting of theJapanese Pharmacological Society, 31 March 1976, Osaka. The authors are grateful to Prof. T. Nambara, Pharmaceutical Institute, Tohoku University, Sendai, for his kind supply of the compounds used in this study.     

Isolation and characterization of fukujusonorone,a 18-norpregnane derivative fromAdonis amurensis Regel et Radd

Zusammenfassung Aus den Wurzeln des japanischen Adonisröschens (Adonis amurensis Regel et Radd) wurde Fukujusonoron, ein neues Aglykon, das sich von 18-nor-Pregnan ableitet, isoliert.

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Abstracted from the Symposium Papers of the 12th Symposium on the Chemistry of Natural Products, Sendai 1968, p. 174. Y.Shimizu, Y.Sato and H.Mitsuhashi, Chem. pharm. Bull., Tokyo15, 2005 (1967); Chem. pharm. Bull., Tokyo, submitted for publication.     

How order and disorder within paramyxoviral nucleoproteins and phosphoproteins orchestrate the molecular interplay of transcription and replication

In this review, we summarize computational and experimental data gathered so far showing that structural disorder is abundant within paramyxoviral nucleoproteins (N) and phosphoproteins (P). In particular, we focus on measles, Nipah, and Hendra viruses and highlight both commonalities and differences with respect to the closely related Sendai virus. The molecular mechanisms that control the disorder-to-order transition undergone by the intrinsically disordered C-terminal domain (N_TAIL) of their N proteins upon binding to the C-terminal X domain (XD) of the homologous P proteins are described in detail. By having a significant residual disorder, N_TAIL–XD complexes are illustrative examples of “fuzziness”, whose possible functional significance is discussed. Finally, the relevance of N–P interactions as promising targets for innovative antiviral approaches is underscored, and the functional advantages of structural disorder for paramyxoviruses are pinpointed.     

DNA fragmentation in leukocytes following repeated low dose sarin exposure in guinea pigs

The objective of this study was to determine levels of DNA fragmentation in blood leukocytes and parietal cortex from guinea pigs following repeated lowlevel exposure to the chemical warfare nerve agent (CWNA) sarin. Guinea pigs were injected (s.c.) once a day for 10 days with saline, or 0.1, 0.2, or 0.4 LD₅₀ (50% mean lethal dose) sarin dissolved in sterile physiological saline. Blood and parietal cortex was collected after injection at 0, 3, and 17 days recovery and evaluated for DNA fragmentation using single-cell gel electrophoresis (Comet assay). Cells were imaged using comet analysis software and three parameters of DNA fragmentation measured: tail length, percent DNA in the tail, and tail moment arm. Repeated low-dose exposure to sarin produced a dose-dependent response in leukocytes at 0 and 3 days post-exposure. There was a significant increase in all measures of DNA fragmentation at 0.2 and 0.4 LD₅₀, but not at 0.1 LD₅₀. There was no significant increase in DNA fragmentation in any of the groups at 17 days post-exposure. Sarin did not produce a systematic dose-dependent response in parietal cortex at any of the time points. However, significant increases in DNA fragmentation at 0.1 and 0.4 LD₅₀ were observed at 0 and 3 days post-exposure. All measures of DNA fragmentation in both leukocytes and neurons returned to control levels by 17 days post-exposure, indicating a small and non-persistent increase in DNA fragmentation following repeated low-level exposure to sarin. Received 23 July 2007; received after revision 23 August 2007; accepted 3 September 2007 Research was conducted in compliance with the Animal Welfare Act, and other Federal statutes and regulation relating to animals and experiments involving animals and adheres to the principles stated in the Guide for the Care and Use of Laboratory Animals, NIH publication 85-23. The views of the authors do not purport to reflect the position of the Department of the Army or the Department of Defense (para 4-3), AR 360–365.     

Effects of araC and aphidicolin on DNA chain elongation rate in HeLa S3 cells

Summary The effects of araC and aphidicolin on DNA chain elongation rate were tested. The rate was markedly reduced at low concentrations. Total DNA synthesis was more inhibited, indicating a role of DNA polymerasea in replicon initiation.     

Relationship between DNA and cholesterol synthesis in kidney after refeeding

Summary DNA and cholesterol synthesis were investigated in the kidneys of fasted-refed rats. Refeeding resulted in an increase in kidney DNA synthesis, as measured by³H-thymidine incorporation, starting at 72 h. The increase in DNA synthesis was accompanied by a stimulation of cholesterol synthesis, as measured by¹⁴C-acetate incorporation into cholesterol.     

Eukaryotic DNA damage checkpoint activation in response to double-strand breaks

Double-strand breaks (DSBs) are the most detrimental form of DNA damage. Failure to repair these cytotoxic lesions can result in genome rearrangements conducive to the development of many diseases, including cancer. The DNA damage response (DDR) ensures the rapid detection and repair of DSBs in order to maintain genome integrity. Central to the DDR are the DNA damage checkpoints. When activated by DNA damage, these sophisticated surveillance mechanisms induce transient cell cycle arrests, allowing sufficient time for DNA repair. Since the term “checkpoint” was coined over 20 years ago, our understanding of the molecular mechanisms governing the DNA damage checkpoint has advanced significantly. These pathways are highly conserved from yeast to humans. Thus, significant findings in yeast may be extrapolated to vertebrates, greatly facilitating the molecular dissection of these complex regulatory networks. This review focuses on the cellular response to DSBs in Saccharomyces cerevisiae, providing a comprehensive overview of how these signalling pathways function to orchestrate the cellular response to DNA damage and preserve genome stability in eukaryotic cells.     

A possibility to achieve genetic transformation in the platyfish-swordtail system (Platypoecilus maculatus — Xiphophorus helleri)

Summary In order to determine how informative homologous donor DNA might be made available to propigment cells of the recipientXiphophorus helleri for transformation, labelled heterologous DNA fromE. coli was injected into the neural crest region or the yolk sac of embryos of the recipient. On the basis of the degradation rate of the donor DNA and the incorporation rate of radioactivity into the recipient DNA, it is concluded that injection into the neural crest region may be a suitable method to make available informative homologous donor DNA for transformation.Supported by DFG through SFB No. 103, and by Stiftung Volkswagenwerk.     

The biochemistry of ancient DNA in bone

The amount of DNA in ancient bone was determined by ethidium bromide staining after the removal of the potent Taq inhibitor, fulvic acid. A complete decalcification and a perfusion protocol were used to recover DNA from bone. A variety of purification techniques including molecular sieve, hydroxyapatite binding and Magic preparations yielded DNA that spanned from 3.4g/g of bone to below detectable limits. Fulvic acid was shown to interfere with the quantification of DNA derived from ancient human skeletal material one hundred to over seven thousand years old. Scanning UV in the 300 to 230 nm range is a simple and sensitive technique for documenting fulvic acid contamination in ancient bone extracts.     

DNA fragmentation in leukocytes following subacute lowdose nerve agent exposure

The objective of the present study was to determine levels of DNA fragmentation in blood leukocytes from guinea pigs by single-cell gel electrophoresis (comet assay) after exposure to the chemical warfare nerve agent (CWNA), soman, at doses ranging from 0.1 LD₅₀ to 0.4 LD₅₀, once per day for either 5 or 10 days. Post-exposure recovery periods ranged from 0 to 17 days. Leukocytes were imaged from each animal, and the images analyzed by computer. Data obtained for exposure to soman demonstrated significant increases in DNA fragmentation in circulating leukocytes in CWNA-treated guinea pigs compared with saline-injected control animals at all doses and time points examined. Notably, significantly increased DNA fragmentation was observed in leukocytes 17 days after cessation of soman exposure. Our findings demonstrate that leukocyte DNA fragmentation assays may provide a sensitive biomarker for low-dose CWNA exposure.Received 29 July 2003; accepted 14 August 2003     

DNA synthesis in exocrine and endocrine pancreas after partial hepatectomy in Syrian golden hamsters

Summary ³H-thymidine autoradiography showed an enhanced DNA synthesis, in acinar and islet cells of pancreas after partial hepatectomy in syrian golden hamsters. A significant nuclear labeling index of acinar cells was observed between 48 and 84 h and reached control levels by 120 h. An increased labeling index of islet cells was also observed, however, this increase was not statistically significant. These results indicate growth factor(s) produced after partial hepatectomy is capable of inducing DNA synthesis in pancreas.This work is supported by NIH grant CA 36043.