ABSTRACT
DNA repair pathways, cell cycle checkpoints, and redox protection systems are essential factors for securing genomic stability. The aim of the present study was to analyze the effect of Ilex paraguariensis (Ip) infusion and one of its polyphenolic components rutin on cellular and molecular damage induced by ionizing radiation. Ip is a beverage drank by most inhabitants of Argentina, Paraguay, Southern Brazil, and Uruguay. The yeast Saccharomyces cerevisiae (SC7Klys 2-3) was used as the eukaryotic model. Exponentially growing cells were exposed to gamma rays (γ) in the presence or absence of Ip or rutin. The concentrations used simulated those found in the habitual infusion. Surviving fractions, mutation frequency, and DNA double-strand breaks (DSB) were determined after treatments. A significant increase in surviving fractions after gamma irradiation was observed following combined exposure to γ+R, or γ+Ip. Upon these concomitant treatments, mutation and DSB frequency decreased significantly. In the mutant strain deficient in MEC1, a significant increase in γ sensitivity and a low effect of rutin on γ-induced chromosomal fragmentation was observed. Results were interpreted in the framework of a model of interaction between radiation-induced free radicals, DNA repair pathways, and checkpoint controls, where the DNA damage that induced activation of MEC1 nodal point of the network could be modulated by Ip components including rutin. Furthermore, ionizing radiation-induced redox cascades can be interrupted by rutin potential and other protectors contained in Ip.
Subject(s)
Antimutagenic Agents/pharmacology , Ilex paraguariensis/chemistry , Plant Extracts/pharmacology , Rutin/pharmacology , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/radiation effects , Cell Survival/drug effects , Cell Survival/radiation effects , Cells, Cultured , Chromatography, Liquid , DNA Breaks, Double-Stranded , DNA Repair , DNA, Fungal/radiation effects , Dose-Response Relationship, Radiation , Gamma Rays , Mass Spectrometry , Mutagenesis , Mutation Rate , Radiation Protection/methods , Reproducibility of ResultsABSTRACT
DNA repair pathways, cell cycle checkpoints, and redox protection systems are essential factors for securing genomic stability. The aim of the present study was to analyze the effect of Ilex paraguariensis (Ip) infusion and one of its polyphenolic components rutin on cellular and molecular damage induced by ionizing radiation. Ip is a beverage drank by most inhabitants of Argentina, Paraguay, Southern Brazil, and Uruguay. The yeast Saccharomyces cerevisiae (SC7Klys 2-3) was used as the eukaryotic model. Exponentially growing cells were exposed to gamma rays (γ) in the presence or absence of Ip or rutin. The concentrations used simulated those found in the habitual infusion. Surviving fractions, mutation frequency, and DNA double-strand breaks (DSB) were determined after treatments. A significant increase in surviving fractions after gamma irradiation was observed following combined exposure to γ+R, or γ+Ip. Upon these concomitant treatments, mutation and DSB frequency decreased significantly. In the mutant strain deficient in MEC1, a significant increase in γ sensitivity and a low effect of rutin on γ-induced chromosomal fragmentation was observed. Results were interpreted in the framework of a model of interaction between radiation-induced free radicals, DNA repair pathways, and checkpoint controls, where the DNA damage that induced activation of MEC1 nodal point of the network could be modulated by Ip components including rutin. Furthermore, ionizing radiation-induced redox cascades can be interrupted by rutin potential and other protectors contained in Ip.
Subject(s)
Rutin/pharmacology , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/radiation effects , Plant Extracts/pharmacology , Antimutagenic Agents/pharmacology , Ilex paraguariensis/chemistry , Radiation Protection/methods , Mass Spectrometry , DNA, Fungal/radiation effects , Cell Survival/drug effects , Cell Survival/radiation effects , Cells, Cultured , Reproducibility of Results , Chromatography, Liquid , Mutagenesis , DNA Repair , Dose-Response Relationship, Radiation , DNA Breaks, Double-Stranded , Mutation Rate , Gamma RaysABSTRACT
We report the results of the seventh edition of the GEP-ISFG mitochondrial DNA (mtDNA) collaborative exercise. The samples submitted to the participant laboratories were blood stains from a maternity case and simulated forensic samples, including a case of mixture. The success rate for the blood stains was moderate ( approximately 77%); even though four inexperienced laboratories concentrated about one-third of the total errors. A similar success was obtained for the analysis of mixed samples (78.8% for a hair-saliva mixture and 69.2% for a saliva-saliva mixture). Two laboratories also dissected the haplotypes contributing to the saliva-saliva mixture. Most of the errors were due to reading problems and misinterpretation of electropherograms, demonstrating once more that the lack of a solid devised experimental approach is the main cause of error in mtDNA testing.
Subject(s)
Artifacts , Clinical Laboratory Techniques/standards , DNA Fingerprinting/standards , DNA, Mitochondrial/genetics , DNA/isolation & purification , Blood Stains , Computer Simulation , DNA/analysis , DNA/genetics , DNA, Mitochondrial/blood , DNA, Mitochondrial/chemistry , Data Interpretation, Statistical , Databases, Factual , Female , Forensic Medicine , Genetic Markers , Hair/chemistry , Haplotypes , Humans , Phylogeny , Polymerase Chain Reaction , Polymorphism, Genetic , Pregnancy , Quality Control , Reference Standards , Saliva/chemistrySubject(s)
Chromosome Aberrations , Prenatal Diagnosis , Chromosomes, Human, Pair 11 , Chromosomes, Human, Pair 2 , Chromosomes, Human, Pair 3 , Chromosomes, Human, Pair 5 , Chromosomes, Human, Pair 9 , Diagnosis, Differential , Female , Genetic Counseling , Humans , In Situ Hybridization, Fluorescence , Infant, Newborn , Male , PregnancyABSTRACT
PURPOSE: We updated an Uruguayan family with hereditary nonpolyposis colorectal cancer first described in 1977, incorporating knowledge of how the hMLH1 germline mutation has been established and shown to segregate in accord with the expected autosomal dominant mode of genetic transmission. METHODS: DNA-based molecular genetic testing was performed in conjunction with genetic counseling. Individuals were provided with their genetic test results, so that at-risk family members would be able to benefit from targeted management programs. RESULTS: We counseled 19 members of this kindred, 13 of whom were positive for the hMLH1 germline mutation. Specific recommendations for surveillance and management were provided. We were able to describe follow-up, including anecdotal cancer survival and pathology findings extending from the initial 1977 report of this family to the present. A remarkable sibship within this kindred was comprised of eight siblings, six of whom underwent resections for colorectal carcinoma between 1963 and 1971. Colon carcinomas before 1977 in this sibship were treated with classic hemicolectomies. Of those who had hemicolectomies for their first primary colorectal cancers, two had a second colon cancer primary, and two had a third colon cancer primary. CONCLUSIONS: Attention given to this extended family with hereditary nonpolyposis colorectal cancer has had a positive impact on the physician community in Uruguay, leading to the identification of additional families with hereditary nonpolyposis colorectal cancer.
Subject(s)
Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , Adaptor Proteins, Signal Transducing , Adult , Aged , Carrier Proteins , Colorectal Neoplasms, Hereditary Nonpolyposis/pathology , Colorectal Neoplasms, Hereditary Nonpolyposis/surgery , Female , Genetic Carrier Screening , Genetic Counseling , Genetic Testing , Germ-Line Mutation/genetics , Humans , Male , Middle Aged , MutL Protein Homolog 1 , Neoplasm Proteins/genetics , Nuclear Proteins , Pedigree , Survival Rate , UruguayABSTRACT
Chinese hamster ovary (CHO) cells were exposed to the restriction endonuclease AluI in the presence of 1.1 M glycerol. Chromosomal aberrations and sister chromatid exchange (SCE) were scored in first post-treatment metaphases following recovery times of 6-18 h. At all recovery times chromosomal aberrations were induced by the enzyme. In AluI-treated damaged cells significant elevations of SCE frequencies were found after recovery times of 12-18 h. These results indicate that SCE, unlike chromosomal aberrations, are induced only in late G1 and early S phase of the cell cycle. The lesions producing SCE are postulated as DNA single-strand breaks and gaps induced by AluI in canonical structures of DNA and in DNA-RNA hybrids.
