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  1. Article ; Online: Episodic overt hepatic encephalopathy after transjugular intrahepatic portosystemic shunt does not increase mortality in patients with cirrhosis.

    Nardelli, Silvia / Riggio, Oliviero / Marra, Fabio / Gioia, Stefania / Saltini, Dario / Bellafante, Daniele / Adotti, Valentina / Guasconi, Tomas / Ridola, Lorenzo / Rosi, Martina / Caporali, Cristian / Fanelli, Fabrizio / Roccarina, Davide / Bianchini, Marcello / Indulti, Federica / Spagnoli, Alessandra / Merli, Manuela / Vizzutti, Francesco / Schepis, Filippo

    Journal of hepatology

    2023  Volume 80, Issue 4, Page(s) 596–602

    Abstract: Background & aims: Overt hepatic encephalopathy (OHE) is a major complication of transjugular intrahepatic portosystemic shunt (TIPS) placement, given its high incidence and possibility of refractoriness to medical treatment. Nevertheless, the impact of ...

    Abstract Background & aims: Overt hepatic encephalopathy (OHE) is a major complication of transjugular intrahepatic portosystemic shunt (TIPS) placement, given its high incidence and possibility of refractoriness to medical treatment. Nevertheless, the impact of post-TIPS OHE on mortality has not been investigated in a large population.
    Methods: We designed a multicenter, non-inferiority, observational study to evaluate the mortality rate at 30 months in patients with and without OHE after TIPS. We analyzed a database of 614 patients who underwent TIPS in three Italian centers and estimated the cumulative incidence of OHE and mortality with competitive risk analyses, setting the non-inferiority limit at 0.12.
    Results: During a median follow-up of 30 months (IQR 12-30), 293 patients developed at least one episode of OHE. Twenty-seven (9.2%) of them experienced recurrent/persistent OHE. Patients with OHE were older (64 [57-71] vs. 59 [50-67] years, p <0.001), had lower albumin (3.1 [2.8-3.5] vs. 3.25 [2.9-3.6] g/dl, p = 0.023), and had a higher prevalence of pre-TIPS OHE (15.4% vs. 9.0%, p = 0.023). Child-Pugh and MELD scores were similar. The 30-month difference in mortality between patients with and without post-TIPS OHE was 0.03 (95% CI -0.042 to 0.102). Multivariable analysis showed that age (subdistribution hazard ratio 1.04, 95% CI 1.02-1.05, p <0.001) and MELD score (subdistribution hazard ratio 1.09, 95% CI 1.05-1.13, p <0.001), but not post-TIPS OHE, were associated with a higher mortality rate. Similar results were obtained when patients undergoing TIPS for variceal re-bleeding prophylaxis (n = 356) or refractory ascites (n = 258) were analyzed separately. The proportion of patients with persistent OHE after TIPS was significantly higher in the group of patients who died. The robustness of these results was increased following propensity score matching.
    Conclusion: Episodic OHE after TIPS is not associated with mortality in patients undergoing TIPS, regardless of the indication.
    Impact and implications: Overt hepatic encephalopathy (OHE) is a common complication in patients with advanced liver disease and it is particularly frequent following transjugular intrahepatic portosystemic shunt (TIPS) placement. In patients with cirrhosis outside the setting of TIPS, the development of OHE negatively impacts survival, regardless of the severity of cirrhosis or the presence of acute-on-chronic liver failure. In this multicenter, non-inferiority, observational study we demonstrated that post-TIPS OHE does not increase the risk of mortality in patients undergoing TIPS, irrespective of the indication. This finding alleviates concerns regarding the weight of this complication after TIPS. Intensive research to improve patient selection and risk stratification remains crucial to enhance the quality of life of patients and caregivers and to avoid undermining the positive effects of TIPS on survival.
    MeSH term(s) Humans ; Hepatic Encephalopathy/epidemiology ; Hepatic Encephalopathy/etiology ; Portasystemic Shunt, Transjugular Intrahepatic/adverse effects ; Quality of Life ; Liver Cirrhosis/complications ; Liver Cirrhosis/surgery ; Hemorrhage/etiology ; Treatment Outcome ; Gastrointestinal Hemorrhage/etiology ; Esophageal and Gastric Varices/etiology
    Language English
    Publishing date 2023-12-13
    Publishing country Netherlands
    Document type Multicenter Study ; Observational Study ; Journal Article
    ZDB-ID 605953-3
    ISSN 1600-0641 ; 0168-8278
    ISSN (online) 1600-0641
    ISSN 0168-8278
    DOI 10.1016/j.jhep.2023.11.033
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Chromatin: the interface between extrinsic cues and the epigenetic regulation of muscle regeneration.

    Guasconi, Valentina / Puri, Pier Lorenzo

    Trends in cell biology

    2009  Volume 19, Issue 6, Page(s) 286–294

    Abstract: Muscle regeneration provides a paradigm by which to study how extrinsic signals coordinate gene expression in somatic stem cells (satellite cells) by directing the genome distribution of chromatin-modifying complexes. Understanding the signal-dependent ... ...

    Abstract Muscle regeneration provides a paradigm by which to study how extrinsic signals coordinate gene expression in somatic stem cells (satellite cells) by directing the genome distribution of chromatin-modifying complexes. Understanding the signal-dependent control of the epigenetic events underlying the transition of muscle stem cells through sequential regeneration stages holds the promise to reveal new targets for selective interventions toward repairing diseased muscles. This review describes the latest findings on how regeneration cues are integrated at the chromatin level to build the transcription network that regulates progression of endogenous muscle progenitors throughout the myogenic program. In particular, we describe how specific epigenetic signatures can confer responsiveness to extrinsic cues on discrete regions of the muscle stem cell genome.
    MeSH term(s) Animals ; Chromatin/physiology ; Epigenesis, Genetic ; Humans ; Muscles/cytology ; Muscles/physiology ; Regeneration
    Chemical Substances Chromatin
    Language English
    Publishing date 2009-04-23
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 30122-x
    ISSN 1879-3088 ; 0962-8924
    ISSN (online) 1879-3088
    ISSN 0962-8924
    DOI 10.1016/j.tcb.2009.03.002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Epigenetic drugs in the treatment of skeletal muscle atrophy.

    Guasconi, Valentina / Puri, Pier Lorenzo

    Current opinion in clinical nutrition and metabolic care

    2008  Volume 11, Issue 3, Page(s) 233–241

    Abstract: Purpose of review: A dynamic network of anabolic and catabolic pathways regulates skeletal muscle mass in adult organisms. Muscle atrophy is the detrimental outcome of an imbalance of this network. The purpose of this review is to provide a critical ... ...

    Abstract Purpose of review: A dynamic network of anabolic and catabolic pathways regulates skeletal muscle mass in adult organisms. Muscle atrophy is the detrimental outcome of an imbalance of this network. The purpose of this review is to provide a critical evaluation of different forms of muscle atrophy from a mechanistic and therapeutic point of view.
    Recent findings: The identification and molecular characterization of distinct pathways implicated in the pathogenesis of muscle atrophy have revealed potential targets for therapeutic interventions. However, an effective application of these therapies requires a better understanding of the relative contribution of these pathways to the development of muscle atrophy in distinct pathological conditions.
    Summary: We propose that the decline in anabolic signals ('passive atrophy') and activation of catabolic pathways ('active atrophy') contribute differently to the pathogenesis of muscle atrophy associated with distinct diseases or unfavorable conditions. Interestingly, these pathways might converge on common transcriptional effectors, suggesting that an optimal intervention should be directed to targets at the chromatin level. We provide the rationale for the use of epigenetic drugs such as deacetylase inhibitors, which target multiple signaling pathways implicated in the pathogenesis of muscle atrophy.
    MeSH term(s) Epigenesis, Genetic ; Gene Expression Regulation ; Genetic Therapy ; Humans ; Muscle Proteins/biosynthesis ; Muscle, Skeletal/enzymology ; Muscle, Skeletal/metabolism ; Muscular Atrophy/prevention & control ; Muscular Atrophy/therapy ; Signal Transduction ; Transcription, Genetic
    Chemical Substances Muscle Proteins
    Language English
    Publishing date 2008-04-10
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 1460178-3
    ISSN 1473-6519 ; 1363-1950
    ISSN (online) 1473-6519
    ISSN 1363-1950
    DOI 10.1097/MCO.0b013e3282fa1810
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Nuclear positioning, gene activity and cancer.

    Guasconi, Valentina / Souidi, Mouloud / Ait-Si-Ali, Slimane

    Cancer biology & therapy

    2005  Volume 4, Issue 2, Page(s) 134–138

    Abstract: In the interphasic nucleus, chromosomes are non-randomly arranged within the nuclear space. Indeed, chromosomes are thought to be organised into "chromosome territories". The size of a chromosome territory is roughly determined by its DNA content, but is ...

    Abstract In the interphasic nucleus, chromosomes are non-randomly arranged within the nuclear space. Indeed, chromosomes are thought to be organised into "chromosome territories". The size of a chromosome territory is roughly determined by its DNA content, but is also influenced by other factors, such as their transcriptional status. Chromatin modifications and positioning of genetic loci in the nucleus play a critical role in the control of gene expression. Emerging evidence suggests that the nucleus is structurally and functionally compartimentalized and desorganization of such a structure might play a major role in the emergence of human diseases such as cancer.
    MeSH term(s) Cell Nucleus/genetics ; Chromosomes, Human/genetics ; Gene Expression Regulation ; Humans ; Neoplasms/genetics ; Neoplasms/metabolism
    Language English
    Publishing date 2005-02-08
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2146305-0
    ISSN 1538-4047
    ISSN 1538-4047
    DOI 10.4161/cbt.4.2.1435
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Chromatin dynamics and cancer.

    Guasconi, Valentina / Ait-Si-Ali, Slimane

    Cancer biology & therapy

    2004  Volume 3, Issue 9, Page(s) 825–830

    Abstract: Chomatin remodeling multi-protein complexes play a key regulatory role in many physiological processes such proliferation and differentiation. The alteration in the recruitment or in the function of one of these chromatin remodeling factors can lead to a ...

    Abstract Chomatin remodeling multi-protein complexes play a key regulatory role in many physiological processes such proliferation and differentiation. The alteration in the recruitment or in the function of one of these chromatin remodeling factors can lead to a proliferation defect, that might result from the aberrant activation or repression of key genes that regulate cell proliferation. In fact, aberrant chromatin remodeling leads to severe human diseases such as leukemia, epithelial cancers and Rubinstein-Taybi syndrome.
    MeSH term(s) Adenosine Triphosphate/metabolism ; Cell Proliferation ; Chromatin/metabolism ; Gene Expression Regulation ; Histones/genetics ; Humans ; Leukemia, Myeloid, Acute/genetics ; Retinoblastoma Protein/genetics
    Chemical Substances Chromatin ; Histones ; Retinoblastoma Protein ; Adenosine Triphosphate (8L70Q75FXE)
    Language English
    Publishing date 2004-09-23
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 2146305-0
    ISSN 1538-4047
    ISSN 1538-4047
    DOI 10.4161/cbt.3.9.1102
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: siRNA as a route to new cancer therapies.

    Ameyar-Zazoua, Maya / Guasconi, Valentina / Ait-Si-Ali, Slimane

    Expert opinion on biological therapy

    2005  Volume 5, Issue 2, Page(s) 221–224

    Abstract: The RNA interference (RNAi) gene-silencing mechanism is induced by double-stranded RNA (dsRNA) and is highly sequence-specific. It is an extremely powerful tool for silencing gene expression in vitro, and might be used as therapy in human pathologies ... ...

    Abstract The RNA interference (RNAi) gene-silencing mechanism is induced by double-stranded RNA (dsRNA) and is highly sequence-specific. It is an extremely powerful tool for silencing gene expression in vitro, and might be used as therapy in human pathologies such as cancer, viral infections and genetic disorders. RNAi was initially discovered in plants, but it has become clear that it is also conserved in animal species. Triggering of RNAi by the introduction of small dsRNA (or small interfering RNA) into living cells as a tool to inhibit the expression of specific genes holds the promise to selectively extinguish the expression of disease-associated genes in humans. On the other hand, RNAi technology will serve to elucidate the functions and interactions of the thousands of human genes in high-throughput systems and help in target validation technology.
    MeSH term(s) Animals ; Genetic Therapy/methods ; Genetic Therapy/trends ; Humans ; Neoplasms/genetics ; Neoplasms/therapy ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/genetics
    Chemical Substances RNA, Small Interfering
    Language English
    Publishing date 2005-02-23
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2052501-1
    ISSN 1744-7682 ; 1471-2598
    ISSN (online) 1744-7682
    ISSN 1471-2598
    DOI 10.1517/14712598.5.2.221
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article: Gamma-ray and hydrogen peroxide induction of gene amplification in hamster cells deficient in DNA double strand break repair.

    Mondello, Chiara / Guasconi, Valentina / Giulotto, Elena / Nuzzo, Fiorella

    DNA repair

    2002  Volume 1, Issue 6, Page(s) 483–493

    Abstract: To investigate the role of DNA double strand breaks (DSBs) and of their repair in gene amplification, we analyzed this process in the V3 Chinese hamster cell line and in the parental line AA8, after exposure to gamma-rays and to hydrogen peroxide (H2O2). ...

    Abstract To investigate the role of DNA double strand breaks (DSBs) and of their repair in gene amplification, we analyzed this process in the V3 Chinese hamster cell line and in the parental line AA8, after exposure to gamma-rays and to hydrogen peroxide (H2O2). V3 is defective in DSB repair because of a mutation in the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) gene, a gene involved in the non-homologous end-joining pathway. As a measure of gene amplification we used the frequency of colonies resistant to N-(phosphonacetyl)-L-aspartate (PALA), since in rodent cells PALA resistance is mainly achieved through the amplification of the CAD (carbamyl-P-synthetase, aspartate transcarbamylase, dihydro-orotase) gene. After treatment with different doses of gamma-rays and of H2O2, we found a dose related increase in the frequency of gene amplification and of chromosome aberrations. When the same doses of damaging agents were used, these increments were higher in V3 than in AA8. These results indicate that DSBs that are not efficiently repaired can be responsible for the induction of gene amplification. H2O2 stimulates gene amplification as well as gamma-rays, however, at similar levels of amplification induction, chromosome damage was about 50% lower. This suggests that gene amplification can be induced by H2O2 through pathways alternative to a direct DNA damage. Stimulation of gene amplification by H2O2, which is one of the products of the aerobic metabolism, supports the hypothesis that cellular metabolic products themselves can be a source of genome instability.
    MeSH term(s) Animals ; Antimetabolites, Antineoplastic/pharmacology ; Aspartic Acid/analogs & derivatives ; Aspartic Acid/pharmacology ; Cell Line ; Chromosome Aberrations/drug effects ; Chromosome Aberrations/radiation effects ; Cricetinae ; Cricetulus ; DNA/biosynthesis ; DNA Damage ; DNA Repair/genetics ; DNA-Activated Protein Kinase ; DNA-Binding Proteins ; Drug Resistance, Neoplasm ; Gamma Rays ; Gene Amplification/drug effects ; Gene Amplification/radiation effects ; Hydrogen Peroxide/pharmacology ; Multienzyme Complexes/genetics ; Oxidants/pharmacology ; Phosphonoacetic Acid/analogs & derivatives ; Phosphonoacetic Acid/pharmacology ; Protein-Serine-Threonine Kinases/deficiency ; Protein-Serine-Threonine Kinases/genetics
    Chemical Substances Antimetabolites, Antineoplastic ; DNA-Binding Proteins ; Multienzyme Complexes ; Oxidants ; Aspartic Acid (30KYC7MIAI) ; sparfosic acid (78QVZ7RG8L) ; DNA (9007-49-2) ; Hydrogen Peroxide (BBX060AN9V) ; DNA-Activated Protein Kinase (EC 2.7.11.1) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; Phosphonoacetic Acid (N919E46723)
    Language English
    Publishing date 2002-12-31
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2071608-4
    ISSN 1568-7856 ; 1568-7864
    ISSN (online) 1568-7856
    ISSN 1568-7864
    DOI 10.1016/s1568-7864(02)00035-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article: Inhibition de l'expression génique par l'ARN interférence : applications dans le domaine du cancer.

    Ait-Si-Ali, Slimane / Guasconi, Valentina / Harel-Bellan, Annick

    Bulletin du cancer

    2004  Volume 91, Issue 1, Page(s) 15–18

    Abstract: Post-transcriptional gene silencing (PTGS) or RNA interference (RNAi) is a powerful tool for silencing gene expression. This mechanism was initially considered as a strange phenomenon limited to few plant species. It has become clear that PTGS occurs in ... ...

    Title translation RNA interference and its possible use in cancer therapy.
    Abstract Post-transcriptional gene silencing (PTGS) or RNA interference (RNAi) is a powerful tool for silencing gene expression. This mechanism was initially considered as a strange phenomenon limited to few plant species. It has become clear that PTGS occurs in both plants and animals and has roles in viral defense and transposon silencing mechanisms. However, the use of RNA interference triggered by the introduction of small double-stranded RNA (dsRNA or siRNA) into mammalian cells as a tool to knock down expression of specific genes holds the promise to selectively inhibit expression of disease-associated genes in humans. On the other hand, there are about 40,000 protein-coding genes in the human genome, but the function of most of them remains unknown. RNAi technology has now been developed for systematically deciphering the functions and interactions of these thousands of genes.
    MeSH term(s) Animals ; Gene Silencing/physiology ; Genetic Therapy/methods ; Humans ; Neoplasm Proteins/genetics ; Neoplasms/therapy ; Plants/genetics ; RNA Interference/physiology ; RNA, Double-Stranded/physiology ; RNA, Small Interfering/physiology
    Chemical Substances Neoplasm Proteins ; RNA, Double-Stranded ; RNA, Small Interfering
    Language French
    Publishing date 2004-01
    Publishing country France
    Document type English Abstract ; Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 213270-9
    ISSN 1769-6917 ; 0007-4551
    ISSN (online) 1769-6917
    ISSN 0007-4551
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Preferential association of irreversibly silenced E2F-target genes with pericentromeric heterochromatin in differentiated muscle cells.

    Guasconi, Valentina / Pritchard, Linda-Louise / Fritsch, Lauriane / Mesner, Larry D / Francastel, Claire / Harel-Bellan, Annick / Ait-Si-Ali, Slimane

    Epigenetics

    2010  Volume 5, Issue 8, Page(s) 704–709

    Abstract: The heterochromatin-associated H3K9 tri-methylase Suv39h1 is involved in the permanent silencing of E2F target genes in differentiating but not in quiescent cells. Here, we tested the hypothesis that permanent silencing of E2F target genes is associated ... ...

    Abstract The heterochromatin-associated H3K9 tri-methylase Suv39h1 is involved in the permanent silencing of E2F target genes in differentiating but not in quiescent cells. Here, we tested the hypothesis that permanent silencing of E2F target genes is associated with their subnuclear positioning close to the pericentromeric heterochromatin compartment, enriched in Suv39h1. Using fluorescence in situ hybridization, we analyzed the subnuclear localization of three E2F target genes relative to the pericentromeric heterochromatin, in cycling fibroblasts or differentiating myoblasts. We observed that all three E2F-target genes have a tendency to relocate closer to the pericentromeric heterochromatin, only when cells differentiate and undergo an irreversible cell cycle withdrawal. These data suggest that repression of E2F target genes in cycling or in differentiating cells is achieved through distinct mechanisms. In differentiating cells, permanent silencing is driven by a Suv39h1-dependent H3K9 tri-methylation and positioning close to the heterochromatin compartment, whereas repression in cycling cells seems independent from subnuclear positioning and requires distinct H3K9 methylation levels.
    MeSH term(s) Cell Differentiation/physiology ; Cell Line ; E2F Transcription Factors/genetics ; E2F Transcription Factors/metabolism ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Gene Silencing/physiology ; Heterochromatin/genetics ; Heterochromatin/metabolism ; Histones/metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Methylation ; Methyltransferases/genetics ; Methyltransferases/metabolism ; Muscle Cells/cytology ; Muscle Cells/metabolism ; Myoblasts/cytology ; Myoblasts/metabolism ; Repressor Proteins/genetics ; Repressor Proteins/metabolism
    Chemical Substances E2F Transcription Factors ; Heterochromatin ; Histones ; Repressor Proteins ; SUV39H1 protein, human (EC 2.1.1.) ; Methyltransferases (EC 2.1.1.-)
    Language English
    Publishing date 2010-11
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1559-2308
    ISSN (online) 1559-2308
    DOI 10.4161/epi.5.8.13025
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Chromatin modification and muscle differentiation.

    Yahi, Hakima / Philipot, Ophélie / Guasconi, Valentina / Fritsch, Lauriane / Ait-Si-Ali, Slimane

    Expert opinion on therapeutic targets

    2006  Volume 10, Issue 6, Page(s) 923–934

    Abstract: Skeletal muscle differentiation is a multistep process, which begins with the commitment of multi-potent mesodermal precursor cells to the muscle fate. These committed cells, the myoblasts, then differentiate and fuse into multinucleated myotubes. The ... ...

    Abstract Skeletal muscle differentiation is a multistep process, which begins with the commitment of multi-potent mesodermal precursor cells to the muscle fate. These committed cells, the myoblasts, then differentiate and fuse into multinucleated myotubes. The final step of muscle differentiation is the maturation of differentiated myotubes into myofibres. Skeletal muscle development requires the coordinated expression of various transcription factors like the members of the myocyte enhancer binding-factor 2 family and the muscle regulatory factors. These transcription factors, in collaboration with chromatin-remodelling complexes, act in specific combinations and within complex transcriptional regulatory networks to achieve skeletal myogenesis. Additional factors involved in the epigenetic regulation of this process continue to be discovered. In this review, the authors discuss the recent discoveries in the epigenetic regulation of myogenesis. They also summarise the role of chromatin-modifying enzymes regulating muscle gene expression. These different factors are often involved in multiple steps of muscle differentiation and have redundant activities. Altogether, the recent findings have allowed a better understanding of myogenesis and have raised new hopes for the pharmacological development of new therapies aimed at muscle degeneration diseases, such as myotonic dystrophy or Duchenne muscular dystrophy.
    MeSH term(s) Animals ; Chromatin/chemistry ; Chromatin/genetics ; Chromatin/metabolism ; Humans ; Muscle Development/drug effects ; Muscle Development/genetics ; Muscle Development/physiology ; Muscle, Skeletal/cytology ; Muscle, Skeletal/drug effects ; Muscle, Skeletal/growth & development ; Muscle, Skeletal/metabolism
    Chemical Substances Chromatin
    Language English
    Publishing date 2006-11-09
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2055208-7
    ISSN 1744-7631 ; 1472-8222
    ISSN (online) 1744-7631
    ISSN 1472-8222
    DOI 10.1517/14728222.10.6.923
    Database MEDical Literature Analysis and Retrieval System OnLINE

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