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11.
Activation of human ribosomal genes by 5-azacytidine 总被引:2,自引:0,他引:2
Cultured human fibroblasts were exposed to 5-azacytidine which inhibits methylation of newly synthesized DNA. A significant increase in the mean number of Ag-stained active nucleolus organizers has been observed in treated cells. This suggests that DNA methylation is involved in modulation of human rRNA synthesis. 相似文献
12.
Nicoletta zini Patrizia Sabatelli Irene Faenza Andrea Ognibene Nadir M. Maraldi 《The Histochemical journal》1996,28(7):495-504
Summary Some key elements of signal transduction have been identified within the nucleus and demonstrated to be responsive to specific
agonists in numerous cell types. In particular, mitogenic stimuli have been reported to induce, a transient increase of the
nuclear phospholipase C β1 activity, causing the release of inositide-derived second messengers, whereas differentiating stimuli induced a decrease
of the enzyme activity and an increase of nuclear phosphatidylinositol 4,5-bisphosphate (PIP2). Recently, we reported evidence, in human osteosarcoma Saos-2 cell lines, on the presence of specific nuclear phospholipase
C isoforms and on the activation of phospholipase C β1 in the nucleus following the exposure to interleukin-1α. In this study we report immunocytochemical ultrastructural evidence on quantitative variations of PIP2 and phospholipase C β1 amounts in the nucleus of Saos-2 cells at different times of exposure to interleukin-1α. After short periods of culture in the presence of the agonist, the intranuclear amount of PIP2 is decreased, while a translocation of phospholipase C β1 occurs from the cytoplasm to the nucleus, in correspondence with the increased hydrolyzing activity of the enzyme. After
longer periods of incubation with interleukin-1α, on the other hand, the intranuclear amount of PIP2 is restored to initial level, while the amount of phospholipase C β1 is increased both at the nuclear and cytoplasmic level, when its activation is no longer effective. The results, compared
with those obtained in other cell types responsive to given agonists, account for a cell-specific modulation of signal transduction
based on polyphosphoinositide breakdown at the nuclear level. 相似文献
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Sara Divari Francesca Valetti Patrizia Caposio Enrica Pessione Maria Cavaletto Ersilia Griva Giorgio Gribaudo Gianfranco Gilardi Carlo Giunta 《European journal of biochemistry》2003,270(10):2244-2253
Phenol hydroxylase (PH) from Acinetobacter radioresistens S13 represents an example of multicomponent aromatic ring monooxygenase made up of three moieties: a reductase (PHR), an oxygenase (PHO) and a regulative component (PHI). The function of the oxygenase component (PHO), here characterized for the first time, is to bind molecular oxygen and catalyse the mono-hydroxylation of substrates (phenol, and with less efficiency, chloro- and methyl-phenol and naphthol). PHO was purified from extracts of A. radioresistens S13 cells and shown to be a dimer of 206 kDa. Each monomer is composed by three subunits: alpha (54 kDa), beta (38 kDa) and gamma (11 kDa). The gene encoding PHO alpha (named mopN) was cloned and sequenced and the corresponding amino acid sequence matched with that of functionally related oxygenases. By structural alignment with the catalytic subunits of methane monooxygenase (MMO) and alkene monooxygenase, we propose that PHO alpha contains the enzyme active site, harbouring a dinuclear iron centre Fe-O-Fe, as also suggested by spectral analysis. Conserved hydrophobic amino acids known to define the substrate recognition pocket, are also present in the alpha-subunit. The prevalence of alpha-helices (99.6%) as studied by CD confirmed the hypothized structural homologies between PHO and MMO. Three parameters (optimum ionic strength, temperature and pH) that affect kinetics of the overall phenol hydroxylase reaction were further analyzed with a fixed optimal PHR/PHI/PHO ratio of 2/1/1. The highest level of activity was evaluated between 0.075 and 0.1 m of ionic strength, the temperature dependence showed a maximum of activity at 24 degrees C and finally the pH for optimal activity was determined to be 7.5. 相似文献
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Long single-stranded DNA molecules have been observed at electron microscope in DNA preparations from synchronized Chinese hamster cells. The amount of single strandedness in parental DNA increases following a prolonged block of DNA synthesis by hydroxyurea as judged by the results obtained using an improved hydroxyapatite chromatography (Hanania et al., 1975). As far as newly replicated DNA is concerned, an increase of the single strand amount has been observed in DNA preparations from cells actively synthesizing DNA. 相似文献
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Effect of Retreatment with Interferon Alone or Interferon plus Ribavirin on Hepatitis C Virus Quasispecies Diversification in Nonresponder Patients with Chronic Hepatitis C 总被引:6,自引:0,他引:6 下载免费PDF全文
Martina Gerotto Daniel G. Sullivan Stephen J. Polyak Liliana Chemello Luisa Cavalletto Patrizia Pontisso Alfredo Alberti David R. Gretch 《Journal of virology》1999,73(9):7241-7247
Alpha interferon (IFN-alpha) treatment is effective on a long-term basis in only 15 to 25% of patients with chronic hepatitis C. The results of recent trials indicate that response rates can be significantly increased when IFN-alpha is given in combination with ribavirin. However, a large number of patients do not respond even to combination therapy. Nonresponsiveness to IFN is characterized by evolution of the hepatitis C virus (HCV) quasispecies. Little is known about the changes occurring within the HCV genomes when nonresponder patients are retreated with IFN or with IFN plus ribavirin. In the present study we have examined the genetic divergence of HCV quasispecies during unsuccessful retreatment with IFN or IFN plus ribavirin. Fifteen nonresponder patients with HCV-1 (4 patients with HCV-1a and 11 patients with HCV-1b) infection were studied while being retreated for 2 months (phase 1) with IFN-alpha (6 MU given three times a week), followed by IFN plus ribavirin or IFN alone for an additional 6 months (phase 2). HCV quasispecies diversification in the E2 hypervariable region-1 (HVR1) and in the putative NS5A IFN sensitivity determining region (ISDR) were analyzed for phase 1 and phase 2 by using the heteroduplex tracking assay and clonal frequency analysis techniques. A major finding of this study was the relatively rapid evolution of the HCV quasispecies observed in both treatment groups during the early phase 1 compared to the late phase 2 of treatment. The rate of quasispecies diversification in HVR1 was significantly higher during phase 1 versus phase 2 both in patients who received IFN plus ribavirin (P = 0.017) and in patients who received IFN alone (P = 0. 05). A trend toward higher rates of quasispecies evolution in the ISDR was also observed during phase 1 in both groups, although the results did not reach statistical significance. However, the NS5A quasispecies appeared to be rather homogeneous and stable in most nonresponder patients, suggesting the presence of a single well-fit major variant, resistant to antiviral treatment, in agreement with published data which have identified an IFN sensitivity determinant region within the NS5A. During the entire 8 months of retreatment, there was no difference in the rate of fixation of mutation between patients who received combination therapy and patients who were treated with IFN alone, suggesting that ribavirin had no major effects on the evolution of the HCV quasispecies after the initial 2 months of IFN therapy. 相似文献
20.
Giovanni Bacci Patrizia Paganin Loredana Lopez Chiara Vanni Claudia Dalmastri Cristina Cantale Loretta Daddiego Gaetano Perrotta Daniela Dolce Patrizia Morelli Vanessa Tuccio Alessandra De Alessandri Ersilia Vita Fiscarelli Giovanni Taccetti Vincenzina Lucidi Annamaria Bevivino Alessio Mengoni 《PloS one》2016,11(6)
Chronic airway infection is a hallmark feature of cystic fibrosis (CF) disease. In the present study, sputum samples from CF patients were collected and characterized by 16S rRNA gene-targeted approach, to assess how lung microbiota composition changes following a severe decline in lung function. In particular, we compared the airway microbiota of two groups of patients with CF, i.e. patients with a substantial decline in their lung function (SD) and patients with a stable lung function (S). The two groups showed a different bacterial composition, with SD patients reporting a more heterogeneous community than the S ones. Pseudomonas was the dominant genus in both S and SD patients followed by Staphylococcus and Prevotella. Other than the classical CF pathogens and the most commonly identified non-classical genera in CF, we found the presence of the unusual anaerobic genus Sneathia. Moreover, the oligotyping analysis revealed the presence of other minor genera described in CF, highlighting the polymicrobial nature of CF infection. Finally, the analysis of correlation and anti-correlation networks showed the presence of antagonism and ecological independence between members of Pseudomonas genus and the rest of CF airways microbiota, with S patients showing a more interconnected community in S patients than in SD ones. This population structure suggests a higher resilience of S microbiota with respect to SD, which in turn may hinder the potential adverse impact of aggressive pathogens (e.g. Pseudomonas). In conclusion, our findings shed a new light on CF airway microbiota ecology, improving current knowledge about its composition and polymicrobial interactions in patients with CF. 相似文献