Assuntos
Animais , Autofagia/fisiologia , Doenças das Aves/parasitologia , Galinhas/parasitologia , Eimeria tenella/fisiologia , Coccidiose/veterinária , Família da Proteína 8 Relacionada à Autofagia/química , Autofagia/genética , Doenças das Aves/prevenção & controle , Marcadores Genéticos/fisiologia , China , Reação em Cadeia da Polimerase , Eimeria tenella/genética , Clonagem Molecular/métodos , Coccidiose/prevenção & controle , Oocistos/isolamento & purificação , Oocistos/fisiologia , Esporozoítos/isolamento & purificação , Esporozoítos/fisiologia , Microscopia Eletrônica de Transmissão , Merozoítos/isolamento & purificação , Merozoítos/fisiologia , Família da Proteína 8 Relacionada à Autofagia/genéticaRESUMO
Abstract Autophagy plays an important role in maintaining cell homeostasis through degradation of denatured proteins and other biological macromolecules. In recent years, many researchers focus on mechanism of autophagy in apicomplexan parasites, but little was known about this process in avian coccidia. In our present study. The cloning, sequencing and characterization of autophagy-related gene (Etatg8) were investigated by quantitative real-time PCR (RT-qPCR), western blotting (WB), indirect immunofluorescence assays (IFAs) and transmission electron microscopy (TEM), respectively. The results have shown 375-bp ORF of Etatg8, encoding a protein of 124 amino acids in E. tenella, the protein structure and properties are similar to other apicomplexan parasites. RT-qPCR revealed Etatg8 gene expression during four developmental stages in E. tenella, but their transcriptional levels were significantly higher at the unsporulated oocysts stage. WB and IFA showed that EtATG8 was lipidated to bind the autophagosome membrane under starvation or rapamycin conditions, and aggregated in the cytoplasm of sporozoites and merozoites, however, the process of autophagosome membrane production can be inhibited by 3-methyladenine. In conclusion, we found that E. tenella has a conserved autophagy mechanism like other apicomplexan parasites, and EtATG8 can be used as a marker for future research on autophagy targeting avian coccidia.
Resumo A autofagia desempenha um papel importante na manutenção da homeostase celular através da degradação de proteínas desnaturadas e outras macromoléculas biológicas. Nos últimos anos, muitos pesquisadores se concentraram no mecanismo da autofagia em parasitas apicomplexos, mas pouco se sabe sobre esse processo na coccidia aviária. No presente estudo, a clonagem, sequenciamento e caracterização de gene relacionado à autofagia Etatg8 foram investigados pela PCR quantitativa em tempo real (RT-qPCR), mancha ocidental (WB), ensaios indiretos de imunofluorescência (IFAs) e microscopia eletrônica de transmissão (TEM), respectivamente. Os resultados mostraram que o gene Etatg8 de E. tenella possui uma ORF de 375 bp, codificando uma proteína de 124 aminoácidos com estrutura e propriedades semelhantes à de outros apicomplexos. RT-qPCR revelou que Etatg8 é expresso durante os quatro estágios de desenvolvimento de E. tenella. Entretanto, seus níveis transcricionais foram significativamente mais elevados na fase de oocisto não esporulados. Os ensaios de manchas ocidental (WB) e de imunofluorescência (IFA) mostraram que a proteína EtATG8 foi lipidada para ligar-se à membrana do autofagossomo sob condições de deficiência nutritiva (em presença de rapamicina) e se agregar no citoplasma de esporozoítas e merozoítas. No entanto, o processo de produção de membrana do autofagossomo pode ser inibido por um inibidor de autofagia (3-meetiladeninatiladenina, 3-MA). Em conclusão, foi demonstrado que E. tenella tem um mecanismo de autofagia conservado, semelhante ao de outros parasitas apicomplexos, e que EtATG8 pode ser usado como um marcador para futuras pesquisas sobre autofagia direcionada à coccidiose aviária.
Assuntos
Animais , Autofagia/fisiologia , Doenças das Aves/parasitologia , Galinhas/parasitologia , Eimeria tenella/fisiologia , Coccidiose/veterinária , Família da Proteína 8 Relacionada à Autofagia/química , Autofagia/genética , Doenças das Aves/prevenção & controle , Marcadores Genéticos/fisiologia , China , Reação em Cadeia da Polimerase , Eimeria tenella/genética , Clonagem Molecular/métodos , Coccidiose/prevenção & controle , Oocistos/isolamento & purificação , Oocistos/fisiologia , Esporozoítos/isolamento & purificação , Esporozoítos/fisiologia , Microscopia Eletrônica de Transmissão , Merozoítos/isolamento & purificação , Merozoítos/fisiologia , Família da Proteína 8 Relacionada à Autofagia/genéticaRESUMO
Protein coding sequences represent only 2% of the human genome. Recent advances have demonstrated that a significant portion of the genome is actively transcribed as non-coding RNA molecules. These non-coding RNAs are emerging as key players in the regulation of biological processes, and act as "fine-tuners" of gene expression. Neurological disorders are caused by a wide range of genetic mutations, epigenetic and environmental factors, and the exact pathophysiology of many of these conditions is still unknown. It is currently recognized that dysregulations in the expression of non-coding RNAs are present in many neurological disorders and may be relevant in the mechanisms leading to disease. In addition, circulating non-coding RNAs are emerging as potential biomarkers with great potential impact in clinical practice. In this review, we discuss mainly the role of microRNAs and long non-coding RNAs in several neurological disorders, such as epilepsy, Huntington disease, fragile X-associated ataxia, spinocerebellar ataxias, amyotrophic lateral sclerosis (ALS), and pain. In addition, we give information about the conditions where microRNAs have demonstrated to be potential biomarkers such as in epilepsy, pain, and ALS.
Assuntos
Humanos , MicroRNAs/fisiologia , RNA Longo não Codificante/fisiologia , Doenças do Sistema Nervoso/genética , Marcadores Genéticos/fisiologia , Regulação da Expressão Gênica , Doenças Neurodegenerativas/genética , MicroRNA Circulante , Doenças Neuromusculares/genéticaRESUMO
OBJECTIVE@#To explore the mutation of Y-STR loci in meiotic allelic transmission in a large pedigree.@*METHODS@#The oral swabs of 163 male individuals were collected from a Lin pedigree. Twenty-two Y-STR genetic markers were typed with AGCU Y24 fluorescent detection kit (AGCU Y24 system), which also contained 16 Y-STR markers included in Yfiler multiple amplification kit (Yfiler system). The genotyping results of Y-STR loci were compared between each two males in the pedigree.@*RESULTS@#There were 20 and 30 kinds of haplotypes obtained with Yfiler and AGCU Y24 systems in 163 male individuals from the Lin pedigree, respectively. The rates referred to haplotype differences (RRHD) of these two typing systems between male pairs were 0.910 5 and 0.922 7, respectively. The average number of marker differences were 6.582 1 and 9.824 8, respectively. The RRHD increased along with the incidents of meiosis.@*CONCLUSION@#Y-STR mutation leads to different Y-STR haplotypes among the male members in a paternal pedigree and the rate of difference increases along with the incidents of meiosis.
Assuntos
Humanos , Masculino , Alelos , Cromossomos Humanos Y/genética , Impressões Digitais de DNA , Ligação Genética , Marcadores Genéticos/fisiologia , Genótipo , Haplótipos , Mutação/genética , LinhagemRESUMO
Barrett's esophagus (BE) is the only known precursor to esophageal adenocarcinoma (EAC), whose incidence has increased sharply in the last 4 decades. The annual conversion rate of BE to cancer is significant, but small. The identification of patients at a higher risk of cancer therefore poses a clinical conundrum. Currently, endoscopic surveillance is recommended in BE patients, with the aim of diagnosing either dysplasia or cancer at early stages, both of which are curable with minimally invasive endoscopic techniques. There is a large variation in clinical practice for endoscopic surveillance, and dysplasia as a marker of increased risk is affected by sampling error and high interobserver variability. Screening programs have not yet been formally accepted, mainly due to the economic burden that would be generated by upper gastrointestinal endoscopy. Screening programs have not yet been formally accepted, mainly due to the economic burden that would be generated by widespread indication to upper gastrointestinal endoscopy. In fact, it is currently difficult to formulate an accurate algorithm to confidently target the population at risk, based on the known clinical risk factors for BE and EAC. This review will focus on the clinical and molecular factors that are involved in the development of BE and its conversion to cancer and on how increased knowledge in these areas can improve the clinical management of the disease.
Assuntos
Animais , Humanos , Camundongos , Adenocarcinoma/etiologia , Esôfago de Barrett/complicações , Diagnóstico por Imagem/métodos , Modelos Animais de Doenças , Epigênese Genética/fisiologia , Neoplasias Esofágicas/diagnóstico , Esofagoscopia/métodos , Previsões , Marcadores Genéticos/fisiologia , Guias de Prática Clínica como Assunto , Fatores de RiscoRESUMO
Maytenus ilicifolia Mart. ex Reis., popularmente conhecida como espinheira-santa, é espécie autóctone pertencente à família Celastraceae, usada para tratamento de úlceras gástricas e gastrites. Devido à importância medicinal, houve aumento no extrativismo das populações naturais, tornando-a uma espécie prioritária para a conservação, a fim de evitar a erosão genética. Buscou-se com este trabalho analisar a diversidade genética de 20 acessos de M. ilicifolia coletados em diferentes localidades no Rio Grande do Sul. Utilizando marcadores moleculares do tipo AFLP, foram testadas oito combinações de primers, que geraram 455 bandas eletroforéticas, 100 por cento polimórficas. As combinações de primers E-ACC/M-CAA, E-ACG/M-CTA, E-ACG/M-CTC apresentaram o maior número de bandas eletroforéticas, 71 cada, totalizando 46,80 por cento do polimorfismo total. Os valores de similaridade genética calculada pelo coeficiente simple matching foram utilizados para gerar o dendrograma de similaridade pelo método UPGMA. Foi obtido alto coeficiente de correlação cofenética (r=0,94), demonstrando elevada representatividade dos dados de similaridade genética no dendrograma. Pela AMOVA verificou-se que 89,33 por cento da diversidade total ocorreram entre indivíduos dentro das populações. A caracterização molecular de acessos de Maytenus ilicifolia por meio de AFLP foi eficiente para identificar diversidade genética. Através da análise de similaridade genética o banco de germoplasma poderia ser composto com os acessos que apresentaram menor similaridade e maior número de alelos, permitindo com que estes fornecessem ampla cobertura do genoma que compõem Maytenus ilicifolia. Os acessos que ficaram agrupados em mesmo cluster e com número reduzido de alelos podem ser descartados deste banco. A diversidade genética intrapopulacional identificada por esse marcador foi muito maior do que aquela entre populações.
Maytenus ilicifolia Mart. ex Reis., popularly known as "espinheira-santa", is an autochthonous species belonging to the Celastraceae family. This species is used to treat ulcers and gastritis. Due to its medicinal importance, the exploitation of natural populations has increased, making the conservation of this species essential to prevent genetic erosion. The aim of this work was to analyze the genetic diversity of 20 M. ilicifolia accessions collected in different localities of Rio Grande do Sul State. Using AFLP-type molecular markers, eight primer combinations were tested, producing 455 electrophoretic profiles, with 100 percent polymorphism. The primer combinations E-ACC/M-CAA, E-ACG/M-CTA and E-ACG/M-CTC presented the largest number of electrophoretic profiles, 71 each, totaling 46.80 percent of the total polymorphism. The values of genetic similarity estimated by Simple Matching Coefficient were used to produce the dendrogram of similarity by the UPGMA method. A high cophenetic correlation coefficient (r = 0.94) was obtained, demonstrating high representativeness of the data of genetic similarity in the dendrogram. Using AMOVA, 89.33 percent of the total diversity were observed among individuals from the same population. The molecular characterization of Maytenus ilicifolia accessions by AFLP allowed the identification of genetic diversity. The genetic similarity analysis indicated that the germplasm bank could be composed of the accessions presenting the lowest similarity and the largest numbers of alleles, providing a comprehensive coverage of Maytenus ilicifolia genome. The accessions that were grouped into one same cluster and with a reduced number of alleles could be disposed of this bank. The genetic diversity identified by this marker within populations was much greater than that between populations.
Assuntos
Brasil , Maytenus/genética , Variação Genética/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Análise de Variância , Modelos Moleculares , Marcadores Genéticos/fisiologia , Marcadores Genéticos/genéticaRESUMO
The garlic cultivars grown in Brazil evolved from somatic mutations and clone selection by breeding programs and by the introduction of germplasm from other countries. Morphological characters have been used to differentiate these cultivars. Two hundred and six random amplified polymorphic DNA markers were utilized for a diversity analysis of the 17 most planted garlic cultivars in Brazil. Bootstrap analysis showed that the number of markers was efficient and sufficient to obtain a coefficient of variation of 10%. Similarity varied between 16 and 98% and cluster analysis showed that, in general, genetic similarities correlate with morphological characters of the cultivars and production cycle variation. High bootstrap values at most of the nodes supported the dendrogram stability. The grouping of most varieties agreed well with previous reports based on morphological characters. As a vegetative-propagated species, viral diseases are a key problem regarding production and quality of the bulbs, causing gradual loss of yield and decrease in storage capacity. To improve the health quality of garlic seed, a virus-free stock of garlic cloves of the Amarante cultivar was obtained. The ability to distinguish garlic cultivars to detect varietal mixing after in vitro multiplication is extremely important, since correct identification is not possible until bulbs are produced. Random amplified polymorphic DNA markers were also used to differentiate cultivars while they are in vitro and not amenable to morphological discrimination. No difference was identified between the fingerprints of the virus-free or of the infected bulks of Amarante, showing that there was no clove mixing in the handling of material in the clonal multiplication phase.
Assuntos
Alho/citologia , Alho/genética , Variação Genética , Produção Agrícola , Alho/classificação , Brasil , Eficiência , Genes de Plantas , Marcadores Genéticos/fisiologia , Fotoperíodo , Filogenia , Controle de Qualidade , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
Hasta tiempos recientes, la periodoncia estuvo orientada a determinar las noxas externas, lo que determinó un oscurecimiento de la participación de otros factores, tales como el genético. El objetivo del presente artículo es brindar al lector una visión de la combinación de la perspectiva molecular de la genética con la de la patogenia de las enfermedades periodontales
Assuntos
Regulação da Expressão Gênica , Doenças Periodontais/genética , Citocinas/fisiologia , Dinoprostona/fisiologia , Predisposição Genética para Doença , Cadeias gama de Imunoglobulina/fisiologia , Imunoglobulina G/imunologia , Interleucina-1/fisiologia , Macrófagos/fisiologia , Marcadores Genéticos/fisiologia , Monócitos/fisiologia , Neutrófilos/fisiologia , Periodontite/genética , Receptores Fc/fisiologia , Fatores de Risco , Fator de Necrose Tumoral alfa/fisiologiaRESUMO
Genetic and environmental factors influence the process of aging. Longevity is extraordinarily constant along species and there are several genes that regulate it. Special consideration deserve apoprotein E alleles, specially allele eee4 that is associated with development of Alzheimer disease, atheroesclerosis and a shorter life. However environmental influences on longevity are of utmost importance. In rats, mice and non human primates, caloric restriction prolongs life and retards the appearance of several conditions associated with aging. This effect of caloric restriction is probably due mainly to a reduction in the generation of primary and secondary reactive oxygen species. Noteworthy is the reduction of DNA oxidative damage and the consequent reduction in transcriptional defects. Telomeres, special structures located in the extremes of chromosomes, are specially susceptible to oxidative damage. This structures have been postulated to act as biological clocks of cells, since their progressive shortening is a signal to stop replication. In humans, telomeres shorten throughout life and in some diseases associated with premature aging such as trisomy 21 and Werner syndrome, the loss of telomeric bases occurs at a higher rate. The knowledge about environmental influences on aging will allow us to increase our life span and to reduce the disabilities associated with aging
Assuntos
Humanos , Animais , Envelhecimento/fisiologia , Necessidades Nutricionais , Ingestão de Energia , Primatas , Envelhecimento/genética , Meio Ambiente , Longevidade , Apolipoproteínas E , Marcadores Genéticos/fisiologia , TelômeroRESUMO
Three highly informative markers genetically linked to Huntington's Disease (HD) were used for diagnosis of HD in Mexican patients, two polymorphic HindIII sites located at D4S10 locus and one VNTR marker at D4S111 locus (VNTR-111). Forty chromosomes from healthy sybjects were tested in order to evaluate the informativeness of the probes. The RFLP HindIII 1 and 2 and the VNTR-111 probes showed a heterozygosity of 0 percent, 45 percent, and 60 percent, respectively. Five families were analyzed, of these, only in two the markers used were informative. In one of them, six membrers showed a decreased risk of inheritance of the mutant gene for Huntington's Didease with 95 percent accuracy
Assuntos
Humanos , DNA , Doenças Genéticas Inatas/terapia , Genética Médica/métodos , Genética Populacional , Doença de Huntington/diagnóstico , Marcadores Genéticos/fisiologia , México , Biologia Molecular , Reação em Cadeia da Polimerase , Fatores de RiscoRESUMO
In order to improve carrier detection of Duchenne and Becker muscular dystrophy, denucleotide sequences repeats (CA) of introns 44, 45, 49 and 50 were used as well as two markers located at the 5' and 3' ends of the dystrophin gene. Haplotypes of the unaffected and affected persons of ten DMD/BMD Mexican families were determined. Fifty eight females were studied, 30 of whom were at-risk STR haplotypes. Furthermore, it was possible to identify a recombination event in the dystrophin gene in one family, and a gonadal mosaicism was found in another family