RESUMO
We compared glyphosate, glufosinate and metsulfuron-methyl to control Ambrosia artemisiifolia under non-crop conditions. A laboratory study showed that A. artemisiifolia is an easy-to-wet species and that glufosinate and glyphosate are quickly absorbed by its leaves (nearly 100% in 24 h). Metsulfuron-methyl absorption was slower (about 50% in 24 h) but was strongly promoted by terpenic alcohol and esterified rapeseed oil. In the greenhouse, all three herbicides were efficacious against A. artemisiifolia, with ED50s of <23, 23 and 0.8 g ha(-1) for glufosinate, glyphosate and metsulfuron-methyl, respectively. These results were confirmed on a non-crop area for glufosinate and glyphosate, which at half the registered dose reached high efficacies at both the 4 to 6-node and flowering stages of A. artemisiifolia. By contrast, metsulfuron-methyl showed no efficacy. However, after treatment at the 4- to 6-node stage, new emergence of A. artemisiifolia led to the presence of vigorous plants that bore numerous flowers and produced high levels of pollen. After treatment at the flowering stage, flower production by A. artemisiifolia was not significantly affected, but achene weight was decreased by 60 to 70% and seed viability was only 8 to 13% for the treated plants, as compared to 85% for the control. No significant difference was observed between the two herbicides and between the doses. It is concluded that glufosinate can be an alternative to glyphosate for the chemical control of A. artemisiifolia on non-crop areas. However, with both herbicides, it is difficult to attain the two objectives of reducing seed production and pollen production by means of only one treatment.
Assuntos
Ambrosia/efeitos dos fármacos , Herbicidas/farmacologia , Herbicidas/farmacocinética , Controle de Pragas/métodos , Absorção , Ambrosia/crescimento & desenvolvimento , Aminobutiratos/farmacocinética , Aminobutiratos/farmacologia , Sulfonatos de Arila/farmacocinética , Sulfonatos de Arila/farmacologia , Relação Dose-Resposta a Droga , Flores/efeitos dos fármacos , Glicina/análogos & derivados , Glicina/farmacocinética , Glicina/farmacologia , Concentração Inibidora 50 , Folhas de Planta/química , Pólen/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Fatores de Tempo , GlifosatoRESUMO
In lower and higher eukaryotes, a family of tightly related proteins designated eRF1 (for eukaryotic release factor 1) catalyses termination of protein synthesis at all three stop codons. The human genome contains four eRF1 homologous sequences localised on chromosomes 5, 6, 7 and X. We report here the cloning and the structural analysis of the human eRF1 gene family. It appears that the gene located on chromosome 5 alone is potentially functional, whereas the other three sequences resemble processed pseudogenes. This is the first description of the structural organisation of the human eRF1 gene, which has been remarkably conserved during evolution and which is essential in the translation termination process.
Assuntos
Cromossomos Humanos Par 5 , Fatores de Terminação de Peptídeos/química , Fatores de Terminação de Peptídeos/genética , Pseudogenes , Sequência de Bases , Cromossomos Humanos Par 6 , Cromossomos Humanos Par 7 , Clonagem Molecular , Cosmídeos , Éxons , Biblioteca Gênica , Humanos , Íntrons , Modelos Genéticos , Dados de Sequência Molecular , Saccharomyces cerevisiae/genética , Cromossomo XRESUMO
Haemochromatosis is an inherited disorder of iron metabolism characterized by a general iron over loading. Without diagnosis and early treatment, it is a serious and potentially fatal disease by cardiac failure or hepatocellular carcinoma in particular. Gene prevalence was estimated at 0.06 in Brittany, so that haemochromatosis may be the most common genetic disease in this area. The biochemical defect of the disease is unknown; only one fact is well established: the iron absorption through duodenal mucosa is excessive. However we don't know if it is a primary event. The gene is also unknown but in 1975 it was located on the short arm of chromosome 6, closely linked to the HLA class I region, less than 1 cM from HLA-A. None of the genes coding for the known iron proteins could be the haemochromatosis gene because of their chromosomal localization. In order to locate this gene with precision, we have used a reverse genetic approach now called positional cloning. Characterization of new polymorphic markers and linkage disequilibrium analysis, have led us to locate the gene within a 350 kb region around HLA-A. We have then searched for all the structural genes in this region. Seven new genes have been so identified and located with precision. A structural analysis of these genes was undertaken to find an eventual abnormality in patients.
Assuntos
Hemocromatose/genética , HumanosAssuntos
Homozigoto , Deficiência da Lecitina Colesterol Aciltransferase/genética , Fosfatidilcolina-Esterol O-Aciltransferase/genética , Mutação Puntual , Adulto , Sequência de Bases , Éxons/genética , Feminino , Humanos , Leucina/genética , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Prolina/genéticaAssuntos
Cromossomos Humanos Par 6 , Genes MHC Classe I , Antígenos HLA-A/genética , Hemocromatose/genética , Mapeamento Cromossômico , Clonagem Molecular , Sondas de DNA , DNA Complementar , Feminino , Expressão Gênica , Biblioteca Gênica , Humanos , Mucosa Intestinal/imunologia , Pulmão/imunologia , Músculo Esquelético/metabolismo , Placenta/imunologia , Gravidez , Pseudogenes , RNA Mensageiro/análise , RNA Mensageiro/biossíntese , Mapeamento por Restrição , Transcrição GênicaRESUMO
Ambrosia artemisiifolia L. (common ragweed), an annual invasive plant, was introduced more than 100 years ago from North America to Europe. Like the majority of other invasive plants in Europe, it develops in open, disturbed areas such as fields, wastelands, roadsides, and riverbanks. Recently, arbuscular mycorrhizal fungi (AMF) have been suspected to play a role in some plant invasion processes. As the common ragweed is known to be colonized by AMF in its native range, the intensity of mycorrhizal root colonization was studied in 35 natural populations in eastern France. About 94% of the A. artemisiifolia populations sampled were mycorrhizal. Root colonization levels varied from 1 to 40% depending on the ecological sites, with lower levels for agricultural habitats and higher levels in disturbed sites, such as wastelands or roadsides. A subsequent greenhouse experiment showed positive impacts of AMF on the growth and development of A. artemisiifolia. It is proposed that the spread of this invasive plant species could be facilitated by AMF, underlining the need to integrate symbiotic interactions in future work on invasive plant processes.
Assuntos
Ambrosia/microbiologia , Ambrosia/fisiologia , Ecossistema , Micorrizas/metabolismo , França , Brotos de Planta , Dinâmica Populacional , Fatores de TempoRESUMO
Numerical simulation is a valuable tool for the study of magnetic susceptibility artifacts from metallic implants. A major difficulty in the simulation lies in the computation of the magnetic field induced by the metallic implant. A new method has been designed and implemented to compute the magnetic field induced by metallic objects of arbitrary shape. The magnetic field is expressed pointwise in terms of a surface integral. Efficient quadrature schemes are proposed to evaluate this integral. Finally, the method is linked to an artifact reconstruction model to simulate the images. Magn Reson Med 45:724-727, 2001.
Assuntos
Imageamento por Ressonância Magnética/métodos , Metais , Próteses e Implantes , Artefatos , Magnetismo , Matemática , Modelos TeóricosRESUMO
A mathematical model of dual-component paramagnetic and diamagnetic material to cancel metal-induced MRI artifacts was developed and verified experimentally. The magnetization produced by metallic material and then the gradient linearity distortion can be cancelled by using such materials with opposing paramagnetic and diamagnetic properties. This concept of dual-component materials provides a novel solution to the problem of MRI artifacts.
Assuntos
Artefatos , Simulação por Computador , Imageamento por Ressonância Magnética/métodos , Magnetismo , Metais , Próteses e Implantes , Imageamento por Ressonância Magnética/instrumentação , Imageamento por Ressonância Magnética/estatística & dados numéricosRESUMO
The authors describe a cystic fibrosis family genotype analysis showing that the R297Q amino acid change is a rare polymorphism rather than a deleterious mutation as previously reported. Indeed in this family two healthy subjects have the following genotypes: delta F508/R297Q and N1303K/R297Q.
Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/genética , Fibrose Cística/genética , Polimorfismo Genético , Adulto , Feminino , Genótipo , Humanos , Masculino , Mutação , LinhagemRESUMO
As part of an effort to characterize the hemochromatosis gene, we selected three non-chimeric yeast artificial chromosomes (YACs) overlapping with the YAC B30 previously described and forming an 800 kilobase contig covering the HLA-A/HLA-F region. The precise physical map of these YACs and of the corresponding genomic region were established. Nine concentrated sites of CpG cutter elements, potentially HTF islands, were mapped. In addition, several probes have been generated as tools for mapping and examining transcripts produced in the region. This allowed for the characterization and localization of two new coding sequences, provisionally named HCG (for hemochromatosis candidate gene) and numbered VIII and IX.
Assuntos
Cromossomos Humanos Par 6 , Hemocromatose/genética , Antígenos de Histocompatibilidade Classe I/genética , Fases de Leitura Aberta , Mapeamento por Restrição , Northern Blotting , Cromossomos Artificiais de Levedura , Clonagem Molecular , Impressões Digitais de DNA , DNA Complementar/genética , Eletroforese em Gel de Campo Pulsado , Biblioteca Gênica , Antígenos HLA/genética , Antígenos HLA-A/genética , Humanos , Dados de Sequência Molecular , Análise de Sequência de DNA , Transcrição GênicaRESUMO
The cystic fibrosis (CF) gene has been cloned and a major mutation identified (delta F508). This 3-bp deletion has been found in approximately 70% of CF chromosomes. We have used the strategy of denaturing gradient gel electrophoresis followed by direct sequencing of the polymerase chain reaction products, in order to detect other mutations in exons 10, 11 and 20 of the CF transmembrane conductance regulator gene. A new mutation, F1286-S, was found in exon 20. It involves a nucleotide change of T-->C at nucleotide 3989 and changes a phenylalanine into serine at position 1286 of the protein.
Assuntos
Fibrose Cística/genética , Éxons , Mutação , Adolescente , Sequência de Bases , Cromossomos Humanos , DNA de Cadeia Simples , Humanos , Lactente , Dados de Sequência Molecular , Polimorfismo de Fragmento de RestriçãoRESUMO
Haemochromatosis is an inherited disorder of iron metabolism characterized by a general iron over loading. Without diagnosis and early treatment, it is a serous and potentially fatal disease by cardiac failure or hepatocellular carcinoma in particular. Gene prevalence was estimated at 0.06 in Brittany, so that haemochromatosis may be the most common genetic disease in this area. The biochemical defect of the disease is unknown; only one fact is well established: the iron absorption through duodenal mucosa is excessive. However, we don't know if it is a primary event. The gene is also unknown but in 1975 it was located on the short arm of chromosome 6, closely linked to the HLA class I region, less than 1 cM from HLA-A. None of the genes coding for the known iron proteins could be the haemochromatosis gene because of their chromosomal localization. In order to locate this gene with precision, we have used a reverse genetic approach now called positional cloning. Characterization of new polymorphic markers and linkage disequilibrium analysis have led us to locate the gene within a 350 kb region around HLA-A. We have then searched for all the structural genes in this region. Seven new genes have been so identified and located with precision. A structural analysis of these genes was undertaken to find an eventual abnormality in patients.
Assuntos
Hemocromatose/genética , Inversão Cromossômica , Mapeamento Cromossômico , Clonagem Molecular/métodos , Frequência do Gene , Triagem de Portadores Genéticos , Antígenos HLA/genética , Antígeno HLA-A1/genética , Hemocromatose/epidemiologia , Hemocromatose/metabolismo , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Desequilíbrio de Ligação , Biologia Molecular , Reação em Cadeia da Polimerase , Polimorfismo Genético , Antígenos HLA-ERESUMO
A yeast artificial chromosome (YAC B30) with a 320 kb insert of genomic DNA which includes the HLA-A gene was used to screen a cDNA library of human duodenal mucosa. Seven cDNA clones were isolated which correspond to seven new non-HLA class I structural genes. These new genes are located within a region that may well contain the gene responsible for hemochromatosis and have therefore been named HCG I-VII (Hemochromatosis Candidate Gene). HCG I, III, V and VI are probably single copy genes, situated at 180, 155, 140 and 230 kb centromeric to HLA-A, respectively. HCG II, IV and VII present several copies: one copy of HCG II, one of HCG IV and one of HCG VII are centromeric to HLA-A (at 30, 70 and 100 kb respectively). Another copy of HCG IV is 20 kb telomeric to HLA-A. Each of the genes localized on the YAC B30 is associated with an CpG/HTF island.
Assuntos
DNA/genética , Genes MHC Classe I , Genes , Antígenos HLA-A/genética , Southern Blotting , Linhagem Celular , Cromossomos Fúngicos , Duodeno , Biblioteca Gênica , Hemocromatose/genética , Humanos , Mucosa Intestinal/imunologia , Especificidade de Órgãos , Mapeamento por Restrição , Saccharomyces cerevisiae/genéticaRESUMO
Positional cloning strategies for the hemochromatosis gene have previously concentrated on a target area restricted to a maximum genomic expanse of 400 kb around the HLA-A and HLA-F loci. Recently, the candidate region has been extended to 2-3 Mb on the distal side of the MHC. In this study, 10 coding sequences [hemochromatosis candidate genes (HCG) I to X] were isolated by cDNA selection using YACs covering the HLA-A/HLA-F subregion. Two of these (HCG II and HCG IV) belong to multigene families, as well as other sequences already described in this region, i.e., P5, pMC 6.7, and HLA class 1. Fingerprinting of the four YACs overlapping the region was performed and allowed partial localization of the different multigene family sequences on each YAC without defining their exact positions. Fingerprinting on cosmids isolated from the ICRF chromosome 6-specific cosmid library allowed more precise localization of the redundant sequences in all of the multigene families and revealed their apparent organization in clusters. Further examination of these intertwined sequences demonstrated that this structural organization resulted from a succession of complex phenomena, including duplications and contractions. This study presents a precise description of the structural organization of the HLA-A/HLA-F region and a determination of the sequences involved in the megabase size polymorphism observed among the A3, A24, and A31 haplotypes.
Assuntos
Antígenos HLA/genética , Antígenos HLA-A/genética , Antígenos de Histocompatibilidade Classe I/genética , Família Multigênica , Linhagem Celular , Mapeamento Cromossômico , Cromossomos Artificiais de Levedura , Clonagem Molecular , Cosmídeos , Impressões Digitais de DNA , Hemocromatose/genética , HumanosRESUMO
The hemochromatosis gene (HFE) maps to 6p21.3 and is less than 1 cM from the HLA class I genes; however, the precise physical location of the gene has remained elusive and controversial. The unambiguous identification of a crossover event within hemochromatosis families is very difficult; it is particularly hampered by the variability of the phenotypic expression as well as by the sex- and age-related penetrance of the disease. For these practical considerations, traditional linkage analysis could prove of limited value in further refining the extrapolated physical position of HFE. We therefore embarked upon a linkage-disequilibrium analysis of HFE and normal chromosomes from the Brittany population. In the present report, 66 hemochromatosis families yielding 151 hemochromatosis chromosomes and 182 normal chromosomes were RFLP-typed with a battery of probes, including two newly derived polymorphic markers from the 6.7 and HLA-F loci located 150 and 250 kb telomeric to HLA-A, respectively. The results suggest a strong peak of existing linkage disequilibrium focused within the i82-to-6.7 interval (approximately 250 kb). The zone of linkage disequilibrium is flanked by the i97 locus, positioned 30 kb proximal to i82, and the HLA-F gene, found 250 kb distal to HLA-A, markers of which display no significant association with HFE. These data support the possibility that HFE resides within the 400-kb expanse of DNA between i97 and HLA-F. Alternatively, the very tight association of HLA-A3 and allele 1 of the 6.7 locus, both of which are comprised by the major ancestral or founder HFE haplotype in Brittany, supports the possibility that the disease gene may reside immediately telomeric to the 6.7 locus within the linkage-disequilibrium zone. Additionally, hemochromatosis haplotypes possessing HLA-A11 and the low-frequency HLA-F polymorphism (allele 2) are supportive of a separate founder chromosome containing a second, independently arising mutant allele. Overall, the establishment of a likely "hemochromatosis critical region" centromeric boundary and the identification of a linkage-disequilibrium zone both significantly contribute to a reduction in the amount of DNA required to be searched for novel coding sequences constituting the HFE defect.
Assuntos
Marcadores Genéticos , Antígenos HLA-A/genética , Haplótipos , Hemocromatose/genética , Desequilíbrio de Ligação , Alelos , Mapeamento Cromossômico , Cromossomos Humanos Par 6 , Feminino , Humanos , MasculinoRESUMO
Congenital bilateral absence of the vas deferens (CBAVD) is found in most males with cystic fibrosis (CF), but this malformation can be observed without any pulmonary or digestive features. We have analyzed 13 exons of the CF gene in a cohort of 25 CBAVD patients. Among the 50 chromosomes studied, 24 mutations were identified: delta F508 (14 cases), R117H (7 cases), R1070W (2 cases), 621 + 1 G --> T (1 case), and A1067V (1 case). Except for delta F508, the most frequent mutations (R117H, R1070W) were not observed in the CF group (109 patients) studied in our laboratory. We discuss the significance of these results.
Assuntos
Fibrose Cística/genética , Proteínas de Membrana/genética , Ducto Deferente/anormalidades , Adulto , Sequência de Bases , Regulador de Condutância Transmembrana em Fibrose Cística , DNA/análise , DNA/química , Éxons , Deleção de Genes , Genótipo , Humanos , Masculino , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Mutação , Reação em Cadeia da Polimerase , Splicing de RNARESUMO
The hemochromatosis gene (HFE) maps to 6p21.3, in close linkage with the HLA Class I genes. Linkage disequilibrium (LD) studies were designed to narrow down the most likely candidate region for HFE, as an alternative to traditional linkage analysis. However, both the HLA-A and D6S105 subregions, which are situated 2-3 cM and approximately 3 Mb apart, have been suggested to contain HFE. The present report extends our previous study based upon the analysis of a large number of HFE and normal chromosomes from 66 families of Breton ancestry. In addition to the previously used RFLP markers spanning the 400-kb surrounding HLA-A, we examined three microsatellites: D6S510, HLA-F, and D6S105. Our combined data not only confirm a peak of LD at D6S105, but also reveal a complex pattern of LD over the i82 to D6S105 interval. Within our ethnically well-defined population of Brittany, the association of HFE with D6S105 is as great as that with HLA-A, while the internal markers display a lower LD. Fine haplotype analysis enabled us to identify two categories of haplotypes segregating with HFE. In contrast to the vast majority of normal haplotypes, 50% of HFE haplotypes are completely conserved over the HLA-A to D6S105 interval. These haplotypes could have been conserved through recombination suppression, selective forces and/or other evolutionary factors. This particular haplotypic configuration might account for the apparent inconsistencies between genetic linkage and LD data, and additionally greatly complicates positional cloning of HFE through disequilibrium mapping.