المؤلفون: Stefano Gambardella, Anna Maria Nardone, Maria Rosaria D'Apice, Giuseppe Novelli, Federica Sangiuolo, Silvia Russo, Mario Bengala, Vincenzina Lucidi

المصدر: BMC Medical Genetics, Vol 5, Iss 1, p 8 (2004)
BMC Medical Genetics

مصطلحات موضوعية: DNA Mutational Analysis, Cystic Fibrosis Transmembrane Conductance Regulator, Nucleic Acid Denaturation, Cystic fibrosis, Male infertility, Cohort Studies, Exon, Leukocytes, Missense mutation, Genetics(clinical), CFTR mutation screening, Polymorphism, Genetic, Exons, Humans, Child, Mutation, Missense, Chromatography, High Pressure Liquid, Italy, Genetic Testing, Cystic Fibrosis, DNA, Introns, Mutation, 5' Untranslated Regions, Sequence Deletion, Genetics (clinical), Genetics, education.field_of_study, Chromatography, Technical Advance, High Pressure Liquid, Allelic heterogeneity, lcsh:Internal medicine, lcsh:QH426-470, Genetic counseling, Population, Biology, Genetic, DHPLC, medicine, Polymorphism, education, lcsh:RC31-1245, medicine.disease, Human genetics, lcsh:Genetics, Settore MED/03 - Genetica Medica, Missense

الوصف: Background Cystic fibrosis (CF) is a multisystem disorder characterised by mutations of the CFTR gene, which encodes for an important component in the coordination of electrolyte movement across of epithelial cell membranes. Symptoms are pulmonary disease, pancreatic exocrine insufficiency, male infertility and elevated sweat concentrations. The CFTR gene has numerous mutations (>1000) and functionally important polymorphisms (>200). Early identification is important to provide appropriate therapeutic interventions, prognostic and genetic counselling and to ensure access to specialised medical services. However, molecular diagnosis by direct mutation screening has proved difficult in certain ethnic groups due to allelic heterogeneity and variable frequency of causative mutations. Methods We applied a gene scanning approach using DHPLC system for analysing specifically all CFTR exons and characterise sequence variations in a subgroup of CF Italian patients from the Lazio region (Central Italy) characterised by an extensive allelic heterogeneity. Results We have identified a total of 36 different mutations representing 88% of the CF chromosomes. Among these are two novel CFTR mutations, including one missense (H199R) and one microdeletion (4167delCTAAGCC). Conclusion Using this approach, we were able to increase our standard power rate of mutation detection of about 11% (77% vs. 88%).

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::648949dbdfe2b4e277a7068dd796e98bTest
http://www.biomedcentral.com/1471-2350/5/8Test

المؤلفون: Dieter C. Gruenert, Monica Lais, Emanuela Bonifazi, Anna Maria Nardone, Annalucia Serafino, Federica Sangiuolo, Emanuela M. Bruscia, Giuseppe Novelli

المصدر: BMC Medical Genetics
BMC medical genetics (Online) 3 (2002): 1–12.
info:cnr-pdr/source/autori:Sangiuolo F., Bruscia E., Serafino A., Nardone A.M., Bonifazi E., Lais M., Gruenert D.C., Novelli G./titolo:In vitro correction of cystic fibrosis epithelial cell lines by small fragment homologous replacement (SFHR) technique./doi:/rivista:BMC medical genetics (Online)/anno:2002/pagina_da:1/pagina_a:12/intervallo_pagine:1–12/volume:3
BMC Medical Genetics, Vol 3, Iss 1, p 8 (2002)

مصطلحات موضوعية: lcsh:Internal medicine, lcsh:QH426-470, Genetic enhancement, Biology, Cystic fibrosis, gene targeting, chemistry.chemical_compound, transmission electron microscopy (TEM), Genetics, medicine, Homologous chromosome, Genetics(clinical), lcsh:RC31-1245, Genetics (clinical), cystic fibrosis transmembrane conductance regulator (CFTR), Gene targeting, Transfection, medicine.disease, gene therapy, Human genetics, In vitro, Cell biology, lcsh:Genetics, transfection, Settore MED/03 - Genetica Medica, chemistry, DNA, Research Article

الوصف: Background SFHR (small fragment homologous replacement)-mediated targeting is a process that has been used to correct specific mutations in mammalian cells. This process involves both chemical and cellular factors that are not yet defined. To evaluate potential of this technique for gene therapy it is necessary to characterize gene transfer efficacy in terms of the transfection vehicle, the genetic target, and the cellular processing of the DNA and DNA-vehicle complex. Methods In this study, small fragments of genomic cystic fibrosis (CF) transmembrane conductance regulator (CFTR) DNA, that comprise the wild-type and ΔF508 sequences, were transfected into immortalized CF and normal airway epithelial cells, respectively. Homologous replacement was evaluated using PCR and sequence-based analyses of cellular DNA and RNA. Individual stages of cationic lipid-facilitated SFHR in cultured cell lines were also examined using transmission electron microscopy (TEM). Results We demonstrated that the lipid/DNA (+/-) ratio influences the mode of entry into the cell and therefore affects the efficacy of SFHR-mediated gene targeting. Lipid/DNA complexes with more negative ratios entered the cell via a plasma membrane fusion pathway. Transfer of the DNA that relies on an endocytic pathway appeared more effective at mediating SFHR. In addition, it was also clear that there is a correlation between the specific cell line transfected and the optimal lipid/DNA ratio. Conclusions These studies provide new insights into factors that underlie SFHR-mediated gene targeting efficacy and into the parameters that can be modulated for its optimization.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5bcd7c8f7c3debfdcbbf1cd5e7735ee2Test
https://doi.org/10.1186/1471-2350-3-8Test

المؤلفون: D'Apice, Maria Rosaria, Novelli, Antonio, di Masi, Alessandra, Biancolella, Michela, Antoccia, Antonio, Gullotta, Francesca, Licata, Norma, Minella, Daniela, Testa, Barbara, Nardone, Anna Maria, Palmieri, Giampiero, Calabrese, Emma, Biancone, Livia, Tanzarella, Caterina, Frontali, Marina, Sangiuolo, Federica, Novelli, Giuseppe, Pallone, Francesco

المصدر: BMC Medical Genetics; 2015, Vol. 16 Issue 1, p1-10, 10p

مصطلحات موضوعية: MALABSORPTION syndromes, MEDICAL genetics, DWARFISM, BODY dysmorphic disorder, DELETION mutation, GENETICS

مستخلص: Background: Copy number variations (CNVs) can contribute to genetic variation among individuals and/or have a significant influence in causing diseases. Many studies consider new CNVs' effects on protein family evolution giving rise to gene duplicates or losses. "Unsuccessful" duplicates that remain in the genome as pseudogenes often exhibit functional roles. So, changes in gene and pseudogene number may contribute to development or act as susceptibility alleles of diseases. Case presentation: We report a de novo heterozygous 271 Kb microdeletion at 8q21.2 region which includes the family of REXO1L genes and pseudogenes in a young man affected by global development delay, progeroid signs, and gastrointestinal anomalies. Molecular and cellular analysis showed that the REXO1L1 gene hemizygosity in a patient's fibroblasts induces genetic instability and increased apoptosis after treatment with different DNA damage-induced agents. Conclusions: The present results support the hypothesis that low copy gene number within REXO1L1 cluster could play a significant role in this complex clinical and cellular phenotype. [ABSTRACT FROM AUTHOR]

: Copyright of BMC Medical Genetics is the property of BioMed Central and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: D'Apice, Maria Rosaria, Gambardella, Stefano, Bengala, Mario, Russo, Silvia, Nardone, Anna Maria, Lucidi, Vincenzina, Sangiuolo, Federica, Novelli, Giuseppe

المصدر: BMC Medical Genetics; 2004, Vol. 5, p8-7, 7p, 3 Charts

مصطلحات موضوعية: CYSTIC fibrosis, GENETIC mutation, CELL membranes, SYMPTOMS, MEDICAL care

مستخلص: Background: Cystic fibrosis (CF) is a multisystem disorder characterised by mutations of the CFTR gene, which encodes for an important component in the coordination of electrolyte movement across of epithelial cell membranes. Symptoms are pulmonary disease, pancreatic exocrine insufficiency, male infertility and elevated sweat concentrations. The CFTR gene has numerous mutations (>1000) and functionally important polymorphisms (>200). Early identification is important to provide appropriate therapeutic interventions, prognostic and genetic counselling and to ensure access to specialised medical services. However, molecular diagnosis by direct mutation screening has proved difficult in certain ethnic groups due to allelic heterogeneity and variable frequency of causative mutations. Methods: We applied a gene scanning approach using DHPLC system for analysing specifically all CFTR exons and characterise sequence variations in a subgroup of CF Italian patients from the Lazio region (Central Italy) characterised by an extensive allelic heterogeneity. Results: We have identified a total of 36 different mutations representing 88% of the CF chromosomes. Among these are two novel CFTR mutations, including one missense (H199R) and one microdeletion (4167delCTAAGCC). Conclusion: Using this approach, we were able to increase our standard power rate of mutation detection of about 11% (77% vs. 88%). [ABSTRACT FROM AUTHOR]

: Copyright of BMC Medical Genetics is the property of BioMed Central and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Sangiuolo, Federica, Bruscia, Emanuela, Serafino, Annalucia, Nardone, Anna Maria, Bonifazi, Emanuela, Lais, Monica, Gruenert, Dieter C., Novelli, Giuseppe

المصدر: BMC Medical Genetics; 2002, Vol. 3, p1-12, 12p, 9 Diagrams, 1 Chart

مصطلحات موضوعية: CYSTIC fibrosis gene, EPITHELIAL cells, GENE targeting, DNA, GENE transfection, GENETIC transformation

مستخلص: Background: SFHR (small fragment homologous replacement)-mediated targeting is a process that has been used to correct specific mutations in mammalian cells. This process involves both chemical and cellular factors that are not yet defined. To evaluate potential of this technique for gene therapy it is necessary to characterize gene transfer efficacy in terms of the transfection vehicle, the genetic target, and the cellular processing of the DNA and DNA-vehicle complex. Methods: In this study, small fragments of genomic cystic fibrosis (CF) transmembrane conductance regulator (CFTR) DNA, that comprise the wild-type and ΔF508 sequences, were transfected into immortalized CF and normal airway epithelial cells, respectively. Homologous replacement was evaluated using PCR and sequence-based analyses of cellular DNA and RNA. Individual stages of cationic lipid-facilitated SFHR in cultured cell lines were also examined using transmission electron microscopy (TEM). Results: We demonstrated that the lipid/DNA (+/-) ratio influences the mode of entry into the cell and therefore affects the efficacy of SFHR-mediated gene targeting. Lipid/DNA complexes with more negative ratios entered the cell via a plasma membrane fusion pathway. Transfer of the DNA that relies on an endocytic pathway appeared more effective at mediating SFHR. In addition, it was also clear that there is a correlation between the specific cell line transfected and the optimal lipid/ DNA ratio. Conclusions: These studies provide new insights into factors that underlie SFHR-mediated gene targeting efficacy and into the parameters that can be modulated for its optimization. [ABSTRACT FROM AUTHOR]

: Copyright of BMC Medical Genetics is the property of BioMed Central and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Giampiero Palmieri, Anna Maria Nardone, Barbara Testa, D. Minella, Livia Biancone, Antonio Novelli, Francesco Pallone, Marina Frontali, Caterina Tanzarella, Michela Biancolella, Alessandra di Masi, Giuseppe Novelli, Emma Calabrese, Antonio Antoccia, Maria Rosaria D'Apice, Norma Licata, Federica Sangiuolo, Francesca Gullotta

المصدر: BMC medical genetics (Online) 16 (2015): 1. doi:10.1186/s12881-015-0164-3
info:cnr-pdr/source/autori:D'Apice M.R.; Novelli A.; di Masi A.; Biancolella M.; Antoccia A.; Gullotta F.; Licata N.; Minella D.; Testa B.; Nardone A.M.; Palmieri G.; Calabrese E.; Biancone L.; Tanzarella C.; Frontali M.; Sangiuolo F.; Novelli G.; Pallone F./titolo:Deletion of REXO1L1 locus in a patient with malabsorption syndrome, growth retardation, and dysmorphic features: A novel recognizable microdeletion syndrome?/doi:10.1186%2Fs12881-015-0164-3/rivista:BMC medical genetics (Online)/anno:2015/pagina_da:1/pagina_a:/intervallo_pagine:1/volume:16

مصطلحات موضوعية: Male, Letter, Adolescent, Apraxias, Pseudogene, Developmental Disabilities, Locus (genetics), Hemizygosity, Biology, Young Adult, Malabsorption Syndromes, Genetics, Humans, Child, Preschool, Gene Expression Regulation, Genetic Loci, Multigene Family, Phenotype, Pseudogenes, Sequence Deletion, Genetics(clinical), Copy-number variation, Allele, Child, Preschool, Gene, Genetics (clinical), cromosoma 8, Microdeletion syndrome, Human genetics, dismorfismo, Settore MED/03 - Genetica Medica, microdelezione

الوصف: Background: Copy number variations (CNVs) can contribute to genetic variation among individuals and/or have a significant influence in causing diseases. Many studies consider new CNVs’ effects on protein family evolution giving rise to gene duplicates or losses. “Unsuccessful” duplicates that remain in the genome as pseudogenes often exhibit functional roles. So, changes in gene and pseudogene number may contribute to development or act as susceptibility alleles of diseases. Case presentation: We report a de novo heterozygous 271 Kb microdeletion at 8q21.2 region which includes the family of REXO1L genes and pseudogenes in a young man affected by global development delay, progeroid signs, and gastrointestinal anomalies. Molecular and cellular analysis showed that the REXO1L1 gene hemizygosity in a patient’s fibroblasts induces genetic instability and increased apoptosis after treatment with different DNA damage-induced agents. Conclusions: The present results support the hypothesis that low copy gene number within REXO1L1 cluster could play a significant role in this complex clinical and cellular phenotype.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::249e481b7cd6f1707c467ca2dadc13deTest

المؤلفون: Nardone Anna, Russo Silvia, Bengala Mario, Gambardella Stefano, D'Apice Maria, Lucidi Vincenzina, Sangiuolo Federica, Novelli Giuseppe

المصدر: BMC Medical Genetics, Vol 5, Iss 1, p 8 (2004)

مصطلحات موضوعية: Cystic fibrosis, CFTR mutation screening, DHPLC, Internal medicine, RC31-1245, Genetics, QH426-470

الوصف: Abstract Background Cystic fibrosis (CF) is a multisystem disorder characterised by mutations of the CFTR gene, which encodes for an important component in the coordination of electrolyte movement across of epithelial cell membranes. Symptoms are pulmonary disease, pancreatic exocrine insufficiency, male infertility and elevated sweat concentrations. The CFTR gene has numerous mutations (>1000) and functionally important polymorphisms (>200). Early identification is important to provide appropriate therapeutic interventions, prognostic and genetic counselling and to ensure access to specialised medical services. However, molecular diagnosis by direct mutation screening has proved difficult in certain ethnic groups due to allelic heterogeneity and variable frequency of causative mutations. Methods We applied a gene scanning approach using DHPLC system for analysing specifically all CFTR exons and characterise sequence variations in a subgroup of CF Italian patients from the Lazio region (Central Italy) characterised by an extensive allelic heterogeneity. Results We have identified a total of 36 different mutations representing 88% of the CF chromosomes. Among these are two novel CFTR mutations, including one missense (H199R) and one microdeletion (4167delCTAAGCC). Conclusion Using this approach, we were able to increase our standard power rate of mutation detection of about 11% (77% vs. 88%).

وصف الملف: electronic resource

العلاقة: http://www.biomedcentral.com/1471-2350/5/8Test; https://doaj.org/toc/1471-2350Test

الوصول الحر: https://doaj.org/article/0fb26338a14448cda963101e178b319eTest

المؤلفون: Bonifazi Emanuela, Nardone Anna, Serafino Annalucia, Bruscia Emanuela, Sangiuolo Federica, Lais Monica, Gruenert Dieter C, Novelli Giuseppe

المصدر: BMC Medical Genetics, Vol 3, Iss 1, p 8 (2002)

مصطلحات موضوعية: gene therapy, cystic fibrosis transmembrane conductance regulator (CFTR), gene targeting, transmission electron microscopy (TEM), transfection, Internal medicine, RC31-1245, Genetics, QH426-470

الوصف: Abstract Background SFHR (small fragment homologous replacement)-mediated targeting is a process that has been used to correct specific mutations in mammalian cells. This process involves both chemical and cellular factors that are not yet defined. To evaluate potential of this technique for gene therapy it is necessary to characterize gene transfer efficacy in terms of the transfection vehicle, the genetic target, and the cellular processing of the DNA and DNA-vehicle complex. Methods In this study, small fragments of genomic cystic fibrosis (CF) transmembrane conductance regulator (CFTR) DNA, that comprise the wild-type and ΔF508 sequences, were transfected into immortalized CF and normal airway epithelial cells, respectively. Homologous replacement was evaluated using PCR and sequence-based analyses of cellular DNA and RNA. Individual stages of cationic lipid-facilitated SFHR in cultured cell lines were also examined using transmission electron microscopy (TEM). Results We demonstrated that the lipid/DNA (+/-) ratio influences the mode of entry into the cell and therefore affects the efficacy of SFHR-mediated gene targeting. Lipid/DNA complexes with more negative ratios entered the cell via a plasma membrane fusion pathway. Transfer of the DNA that relies on an endocytic pathway appeared more effective at mediating SFHR. In addition, it was also clear that there is a correlation between the specific cell line transfected and the optimal lipid/DNA ratio. Conclusions These studies provide new insights into factors that underlie SFHR-mediated gene targeting efficacy and into the parameters that can be modulated for its optimization.

وصف الملف: electronic resource

العلاقة: http://www.biomedcentral.com/1471-2350/3/8Test; https://doaj.org/toc/1471-2350Test

الوصول الحر: https://doaj.org/article/fd8727020bee45af8befcf80f2c2f0edTest