RESUMO
Lipoprotein lipase is essential for triglyceride hydrolysis. The polymorphisms S447X in exon 9 and HindIII in intron 8 have been associated with lower triglyceride levels and lower cardiovascular risk in adult men. We examined the association of these lipoprotein lipase polymorphisms with high-density lipoprotein (HDL) and triglyceride levels in elderly men. Blood samples were obtained from 87 elderly men, 48 of whom had cardiovascular disease and 39 (controls) had no history of cardiovascular events. The lipoprotein lipase polymorphisms were analyzed by PCR-RFLP. Allele frequencies were H- = 27.9% and X = 21.5%. There were no significant differences in allele frequencies or blood lipid levels between cardiovascular disease and control groups. However, the X allele was associated with a lower triglyceride/HDL ratio, 2.30 vs 3.02 for X allele absent (P = 0.03); the H-X haplotype was associated with lower triglyceride levels compared to the H+S haplotype (1.22 vs 1.58 mM, respectively) and a lower triglyceride/HDL ratio (2.29 vs 3.26, respectively). The X allele and H-X haplotype were associated with lower triglyceride/HDL ratios in these elderly men, independent of the history of cardiovascular events.
Assuntos
Lipase Lipoproteica/genética , Lipoproteínas HDL/sangue , Triglicerídeos/sangue , Idoso , Idoso de 80 Anos ou mais , Brasil , Desoxirribonuclease HindIII/química , Éxons , Frequência do Gene , Genes Ligados ao Cromossomo X , Haplótipos , Humanos , Íntrons , Masculino , Polimorfismo GenéticoRESUMO
Plasminogen activator inhibitor type 1 (PAI-1) is an inhibitor of plasmin production. Plasmin can directly or indirectly to degrade cartilage and bone matrix. The PAI-1 HindIII polymorphism has been associated with high PAI-1 plasma levels in myocardial infarction patients and control populations. Furthermore, it has been associated with the angiographic extent of coronary artery disease, but their involvement in other diseases is still uncertain. Here, we assessed the relationship between PAI-1 HindIII polymorphism and PAI-1 plasma levels in rheumatoid arthritis (RA). One hundred and twenty-five RA patients and 132 control subjects (CS) were included. Genotypes were identified by the polymerase chain reaction-restriction fragment length polymorphism technique and PAI-1 plasma levels were quantified using an ELISA kit. Not significant differences in genotype and allele frequencies between both studied groups were observed (P > 0.05). RA patients showed lower PAI-1 plasma levels (18.92 +/- 12.94 ng/ml) than CS (23.68 +/- 23.38 ng/ml), without significant difference (P = 0.299). However, in RA patients the C/G genotype carriers showed higher PAI-1 plasma levels (23.00 +/- 13.81 ng/ml) with respect to C/C (16.77 +/- 11.97 ng/ml) and G/G (10.47 +/- 7.07 ng/ml) genotype carriers (P = 0.036). The PAI-1 HindIII polymorphism was not associated with RA susceptibility. However, the C/G genotype is associated with high PAI-1 plasma levels in RA patients.
Assuntos
Artrite Reumatoide/patologia , Inibidor 1 de Ativador de Plasminogênio/sangue , Inibidor 1 de Ativador de Plasminogênio/genética , Polimorfismo Genético , Desoxirribonuclease HindIII/metabolismo , Suscetibilidade a Doenças , Ensaio de Imunoadsorção Enzimática/métodos , Frequência do Gene , Humanos , Plasma/química , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de RestriçãoRESUMO
Microsporum canis is the main zoophylic dermatophyte in dogs and cats, and it is also an important zoonotic agent. The literature showed that cats are asymptomatic carriers of M. canis. This is apparently due to host resistance and/or the presence of strains with lower virulence. This study was aimed to evaluate the keratinolytic, elastinolytic and collagenolytic activities of M. canis strains and their relationship with symptomatic and asymptomatic cats. In addition, these strains were analysed by RFLP. The strains isolated from cats with clinical dermatophytosis had higher keratinase and elastase activity than those isolated from asymptomatic animals (p minus than 0.05). There were not differences in RFLP patterns based on Hind III digestion.
Assuntos
Doenças do Gato/microbiologia , Gatos/microbiologia , Colagenases/isolamento & purificação , Proteínas Fúngicas/isolamento & purificação , Microsporum/isolamento & purificação , Elastase Pancreática/isolamento & purificação , Peptídeo Hidrolases/isolamento & purificação , Tinha/veterinária , Animais , Colagenases/metabolismo , DNA Fúngico/genética , Desoxirribonuclease HindIII , Líquido Extracelular/enzimologia , Microsporum/enzimologia , Microsporum/genética , Microsporum/patogenicidade , Elastase Pancreática/metabolismo , Peptídeo Hidrolases/metabolismo , Polimorfismo de Fragmento de Restrição , Especificidade da Espécie , Tinha/microbiologia , VirulênciaRESUMO
BACKGROUND: Lipoprotein lipase has an important role in lipid metabolism. Elevated levels of very-low-density lipoprotein (VLDL) and low-density lipoprotein (LDL) are associated with increased risk of coronary artery disease and low levels of high-density lipoprotein (HDL) are potentially atherogenic. The HindIII and S447X polymorphisms of the lipoprotein lipase (LPL) gene are associated with cardiovascular disease in some populations. METHODS: LPL HindIII and S447X polymorphisms were analyzed in 343 individuals of 66-97 years of age from a cohort of a Brazilian elderly longitudinal study. Allele frequencies, genotype distribution and allele association with major morbidities and with serum lipid, urea, creatinine and albumin levels were also investigated. The whole sample was genotyped by PCR-restriction fragment length polymorphism (RFLP). Descriptive statistics, logistic regression analysis and t-test were used. RESULTS: Allele frequencies were H(+)=0.652 and H(-)=0.348 for LPL HindIII and S=0.824 and X=0.176 for LPL S447X polymorphism. Both polymorphisms have frequencies similar to those in some European populations. LPL HindIII polymorphism showed significant association of the H(+) allele with myocardial infarction. The H(-) allele showed a tendency to associate with higher HDL levels. The LPL S447X S allele was associated with higher triglyceride levels. CONCLUSIONS: These findings may help to identify risk factors for subjects and families and clarify the physiopathological role of these polymorphisms in age-related diseases.
Assuntos
Lipídeos/sangue , Lipase Lipoproteica/genética , Infarto do Miocárdio/genética , Polimorfismo Genético/fisiologia , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Estudos de Coortes , Desoxirribonuclease HindIII , Frequência do Gene , Predisposição Genética para Doença , Humanos , Metabolismo dos Lipídeos/genética , Estudos Longitudinais , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/etiologia , Fatores de Risco , Triglicerídeos/sangueRESUMO
Autosomal dominant facioscapulohumeral muscular dystrophy (FSHD) is associated with contractions of D4Z4 repeat on 4q35. It displays a remarkable inter- and intra-familial clinical variability ranging from severe phenotype to asymptomatic carriers. Mosaicism for the contracted FSHD-sized allele is a recurrent finding, but only DNA from lymphocytes had been studied. It is currently not known if mosaicism is unequally distributed between different tissues and if muscle is relatively spared for the presence of the disease allele in mosaic asymptomatic carriers of a disease allele. Here we compare DNA extracted from peripheral blood lymphocytes (PBL), fibroblasts and muscle from a mosaic asymptomatic female carrier and mother of a FSHD patient. PFGE analysis showed a complex allelic segregation: two independent mitotic rearrangement episodes occurred, resulting in mosaicism for a contracted D4Z4 repeat on 4q35 in the mother and mosaicism for an expanded D4Z4 repeat on 10q26 in the affected daughter. The results show that the proportion of mosaicism in PBL and muscle were comparable, while in fibroblasts there was some variation in the mosaicism, which might be caused by culturing artefacts. This finding supports the hypothesis that a mitotic contraction of D4Z4 is an early embryonic event and indicates that the degree of mosaicism in PBL is representative for that of muscle.
Assuntos
Mosaicismo , Distrofia Muscular Facioescapuloumeral/patologia , Sequências Repetitivas de Ácido Nucleico/genética , Alelos , Cromossomos Humanos Par 4 , DNA/genética , DNA/metabolismo , Desoxirribonuclease EcoRI/metabolismo , Desoxirribonuclease HindIII/metabolismo , Eletroforese em Gel de Campo Pulsado , Saúde da Família , Feminino , Fibroblastos/metabolismo , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Músculos/metabolismo , Distrofia Muscular Facioescapuloumeral/sangue , Distrofia Muscular Facioescapuloumeral/genética , LinhagemRESUMO
Se mostró la caracterización genómica de la cepa utilizada en la producción del biolarvicida Bactivec. Se compararon los patrones de digestión de la cepa de referencia 266 de Bacillus thuringiensis de la colección del Instituto Pushkin del antiguo Leningrado y 2 cepas aisladas de 2 lotes del biolarvicida. Las digestiones del ADN cromosomal se realizaron con las enzimas HindIII y EcoRI. En todos los casos se observó el mismo patrón de restricción, lo que muestra la estabilidad genética del ingrediente activo del biolarvicida Bactivec
Assuntos
Bacillus thuringiensis , Controle de Doenças Transmissíveis , Desoxirribonuclease EcoRI , Desoxirribonuclease HindIII , Vetores de Doenças , Inseticidas , Controle de QualidadeRESUMO
Single-enzyme amplified fragment length polymorphism (SE-AFLP) analyses were used to differentiate 97 isolates of porcine Pasteurella multocida subsp. multocida. The strains, isolated from animals with pneumonia, rhinitis, and septicemia, were classified as capsular types A, D, and F. SE-AFLP showed a discriminatory index of 0.87 and identified 18 different profiles.
Assuntos
Desoxirribonuclease HindIII/metabolismo , Infecções por Pasteurella/veterinária , Pasteurella multocida/classificação , Polimorfismo de Fragmento de Restrição , Doenças dos Suínos/microbiologia , Animais , Bacteriemia/microbiologia , Bacteriemia/veterinária , Cápsulas Bacterianas/imunologia , Toxinas Bacterianas/genética , Técnicas de Tipagem Bacteriana , Infecções por Pasteurella/microbiologia , Pasteurella multocida/genética , Pasteurella multocida/isolamento & purificação , Pneumonia Bacteriana/microbiologia , Pneumonia Bacteriana/veterinária , Reação em Cadeia da Polimerase , Rinite Atrófica/microbiologia , Rinite Atrófica/veterinária , SuínosRESUMO
Outbreaks of an epidermic disease suggesting parapox virus infections have been observed in all major herds of sheep and goats from different geographical areas of Brazil. Clinical samples (dried scabs) were collected and orf virus was isolated and characterized by electron microscopy in previous work. In order to characterize these viruses at the molecular level, a modified methodology for genomic DNA extraction directly from scabs was used and such DNA was used to derive the restriction enzyme digestion patterns for clinical samples from three distinct geographic origins. Pulsed field gel electrophoresis was used to separate restriction enzyme DNA fragments and heterogeneity among isolates from different geographic areas could be observed on stained gels. The HindIII-G DNA fragment from orf-A virus genome was cloned and hybridized to DNA of other orf virus isolates. Further heterogeneity was confirmed by these hybridizations.
Assuntos
Ectima Contagioso/virologia , Doenças das Cabras/virologia , Vírus do Orf/genética , Animais , Brasil/epidemiologia , Clonagem Molecular , Sondas de DNA/química , DNA Viral/química , DNA Viral/isolamento & purificação , Desoxirribonuclease HindIII/química , Surtos de Doenças/veterinária , Ectima Contagioso/epidemiologia , Eletroforese em Gel de Ágar/veterinária , Eletroforese em Gel de Campo Pulsado/veterinária , Doenças das Cabras/epidemiologia , Cabras , Microscopia Eletrônica/veterinária , Hibridização de Ácido Nucleico , Vírus do Orf/química , Vírus do Orf/classificação , OvinosRESUMO
The folate-sensitive fragile site FRAXE is located in proximal Xq28 of the human X chromosome and lies approximately 600 kb distal to the fragile X syndrome (FRAXA) fragile site at Xq27.3. Although FRAXA and FRAXE are indistinguishable by means of conventional cytogenetics, they can now be delineated at the molecular level and provides the basis for a proper diagnosis. The screening for CGG amplifications in the FMR1 gene was based on standard protocols using EcoRI digests on Southern blots and hybridization with the StB12.3 probe. The FRAXE mutation was analyzed by digestion with HindIII and the filters were probed with OxE20. We present the results of 144 patients referred for fragile X testing but negative for the FMR1 gene trinucleotide expansion, that were also screened for the FMR2 expansion. For FRAXE mutation a molecular protocol for OxE18 probe was used, in the DNA samples digested with EcoRI on the same blots as those used for detection of FRAXA. None of the patients tested were positive for the FRAXE expansion. This technique was successfully established into our laboratory routine showing the practical use of testing for FRAXA and FRAXE in a large series of patients.
Assuntos
Sondas de DNA/genética , Síndrome do Cromossomo X Frágil/genética , Deficiência Intelectual/genética , Proteínas Nucleares , Proteínas de Ligação a RNA , Transativadores , Southern Blotting , Brasil , Desoxirribonuclease EcoRI , Desoxirribonuclease HindIII , Feminino , Proteína do X Frágil da Deficiência Intelectual , Síndrome do Cromossomo X Frágil/diagnóstico , Testes Genéticos , Humanos , Masculino , Mutação , Proteínas do Tecido Nervoso/genética , Proteínas/genética , Repetições de TrinucleotídeosRESUMO
In this paper, we report a fast, simple, and reproducible staining protocol for nucleic acids in agarose gels with a sensitivity in the order of 10 pg/mm2. It took only three steps: fixation, incubation with silver ions, and development of the gels (total time 50 min). The resulting calibration curves (area vs ng of loaded DNA) after a densitometric scanning of agarose gels stained with this procedure were linear up to 50 ng of double-stranded DNA. We found this method suitable for routine laboratory use and especially appropriate for densitometric analysis due to homogeneous background development. Furthermore, it avoids the pretreatment and/or drying steps proposed by other authors for agarose gels.