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2.
Braz. dent. j ; 28(2): 148-151, mar.-Apr. 2017. tab, graf
Artigo em Inglês | LILACS (Américas) | ID: biblio-839142

RESUMO

Chromosomal instability, leading to aneuploidy, is one of the hallmarks of human cancers. USP44 (ubiquitin specific peptidase 44) is an important molecule that plays a regulatory role in the mitotic checkpoint and USP44 loss causes chromosome mis-segregation, aneuploidy and tumorigenesis in vivo. In this study, it was investigated the immunoexpression of USP44 in 28 malignant salivary gland neoplasms and associated the results with DNA ploidy status assessed by image cytometry. USP44 protein was widely expressed in most of the tumor samples and no clear association could be established between its expression and DNA ploidy status or tumor size. On this basis, it may be concluded that the aneuploidy of the salivary gland cancers included in this study was not driven by loss of USP44 protein expression.


Resumo Instabilidade cromossômica acarretando aneuploidia é um dos fatores marcantes de neoplasias malignas humanas. USP44 (peptidase específica de ubiquitina 44) é uma importante molécula que exerce um papel regulador no ciclo celular e sua perda pode acarretar em segregação cromossômica deficiente, aneuploidia e desenvolvimento de tumores in vivo. Neste estudo, investigou-se a expressão imuno-histoquímica da proteína USP44 em 28 neoplasias malignas de glândulas salivares, associando-se os resultados com o estado de ploidia do DNA avaliado por citometria de fluxo. A proteína USP44 apresentou ampla expressão na maioria das amostras avaliadas e não foi observada associação entre a expressão protéica e o estado de ploidia do DNA ou extensão do tumor. Baseando-se nos resultados, concluiu-se que a aneuploidia das neoplasias malignas de glândulas de salivares incluídas neste estudo não foi influenciada pela perda de expressão da proteína USP44.


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Aneuploidia , DNA/genética , Neoplasias das Glândulas Salivares/genética , Proteases Específicas de Ubiquitina/metabolismo
3.
Rev. cuba. invest. bioméd ; 35(2): 184-194, abr.-jun. 2016.
Artigo em Espanhol | LILACS (Américas), CUMED | ID: lil-783764

RESUMO

Desde hace varias décadas el ensayo Cometa, o electroforesis alcalina de células individuales, se ha convertido en un método establecido para el estudio del daño de ácido desoxirribonucleico, con múltiples aplicaciones en ensayos de genotoxicidad, estudios de biomonitoreo en humanos, epidemiologia molecular y ecotoxicología; así como una herramienta fundamental para investigaciones sobre daño y reparación del ácido desoxirribonucleico. Este ensayo se distinguió por su simplicidad, sensibilidad, versatilidad, rapidez y economía. Es una poderosa técnica que se basa en la visualización microscópica de las imágenes del ácido desoxirribonucleico después que las células son embebidas en agarosa, lisadas y sometidas a una electroforesis alcalina. Esta metodología básica ha sido ampliada, y permite ahora, detectar con alta sensibilidad una gran variedad de daños del ácido desoxirribonucleico en cualquier tipo de células. La inclusión en este ensayo, de enzimas capaces de producir lesiones específicas en la hebra de ácido desoxirribonucleico, ha incrementado su rango de detección y sensibilidad. Pero es importante tener claro que su especificidad no es absoluta. El propósito es destacar algunos aspectos útiles de este método y sus ventajas; describir la experiencia en algunos aspectos técnicos del proceder, normalizado según las condiciones del laboratorio en el instituto para ampliar su utilización en el país.


For several decades now the comet assay (single cell gel electrophoresis assay) has been the method used for the study of deoxyribonucleic acid damage, with multiple applications in genotoxicity assays, biomonitoring studies in humans, molecular epidemiology and ecotoxicology, and a fundamental tool for research into deoxyribonucleic acid damage and repair. The comet assay has stood out for its simplicity, sensitivity, versatility, rapidity and economy. It is a powerful technique based on microscopic visualization of deoxyribonucleic acid images after the cells have been embedded in agarose, lysed and subjected to alkaline electrophoresis. This basic methodology has been broadened, and may now detect with great sensitivity a large variety of deoxyribonucleic acid damage in any type of cell. Inclusion in the assay of enzymes capable of producing specific lesions on the deoxyribonucleic strand has broadened its detection range and sensitivity. However, it is important to bear in mind that its specificity is not absolute. The purpose of the present study is to point out some useful aspects and advantages of the method, and describe the experience with some technical aspects of the procedure, standardized in keeping with the conditions in the laboratory at the institute to extend its use in the country.


Assuntos
Humanos , DNA/genética , Ensaio Cometa/métodos
4.
São Paulo; s.n; s.n; 2015. 133 p. tab, graf, ilus.
Tese em Português | LILACS (Américas) | ID: biblio-847321

RESUMO

O tratamento padrão para pacientes com câncer de reto localmente avançado consiste no uso de quimioradioterapia neoadjuvante (QRTn), seguida por cirurgia. Uma fração significativa dos pacientes responde completamente ao tratamento e no momento da reavaliação não apresenta evidência clínica nem radiológica de doença. Uma abordagem alternativa, Watch and Wait, propõe não operar imediatamente esses pacientes e submetê-los a um protocolo de observação frequente, a fim de evitar as morbidades associadas à cirurgia. No entanto, a avaliação da resposta ao tratamento ainda é um desafio, devido à subjetividade da avaliação clínica e a ausência de exames radiológicos suficientemente sensíveis e específicos para garantir a ausência de células tumorais residuais ou capazes de detectar a recorrência precoce da doença. DNA circulante contendo alterações genéticas específicas do tumor (ctDNA) pode ser encontrado na fração livre de células do sangue e tem sido utilizado para monitorar a dinâmica tumoral em tumores sólidos. Avanços recentes das tecnologias de sequenciamento permitem a identificação eficiente e rápida e a um custo relativamente baixo de alterações genéticas em tumores individuais, superando o problema imposto pela ausência de alterações genéticas recorrentes nesses tumores. Essas alterações podem ser utilizadas como biomarcadores personalizados para monitorar a resposta ao tratamento, detectar doença residual e a recidiva precoce do tumor. O objetivo deste trabalho foi identificar e estudar biomarcadores personalizados em pacientes com câncer de reto localmente avançado tratados com QRTn e avaliar a capacidade desses biomarcadores para monitorar a dinâmica tumoral, e auxiliar na definição da conduta cirúrgica e na detecção da recidiva precoce da doença. Biópsias de seis pacientes com adenocarcinoma de reto distal (cT2- 3N0-1M0), foram coletadas prospectivamente pré-tratamento. O DNA genômico extraído a partir das biópsias foi usado para construir bibliotecas tipo mate-pair para o sequenciamento do genoma completo, utilizando a plataforma SOLiD. Rearranjos inter e intracromossômicos foram identificados utilizando programas computacionais desenvolvidos pelo nosso grupo de pesquisa e em seguida foram validados utilizando PCR e sequenciamento Sanger. Foram validadas, pelo menos, três variações estruturais para cada paciente. Amostras de plasma foram coletadas no momento do diagnóstico, depois da QRTn e durante o seguimento. DNA circulante total foi extraído a partir das amostras de plasma e ensaios personalizados foram desenvolvidos para monitorar a presença de variações estruturais através de PCR Digital. ctDNA foi detectado em todas amostras de plasma pré-tratamento de pacientes com tumores T3. A detecção desses biomarcadores apresentou boa correlação com a resposta ao tratamento, no entanto, esta abordagem não foi sensível o suficiente para detectar doença residual. Para dois pacientes que desenvolveram doença metastática foi verificado um aumento nos níveis de ctDNA com pelo menos 36 semanas antes do diagnóstico clínico de doença metastática, sendo possível correlacionar os níveis de ctDNA detectados em coletas subsequentes com a resposta ao tratamento sistêmico de segunda linha. Este estudo, embora de caráter exploratório, gerou dados relevantes e suficientes para justificar a realização de estudos adicionais para avaliar a aplicação dos biomarcadores personalizados na definição da conduta cirúrgica e no acompanhamento de pacientes com câncer de reto tratados com QRTn


The standard treatment for patients with locally advanced rectal cancer comprises in neoadjuvant chemo radiotherapy (nCRT), followed by surgery. A significant fraction of these patients show complete response to the treatment and at the time of reassessment, there are no clinical and nor radiological evidence of residual tumor. An alternative approach, Watch and Wait, proposes not to immediately operate these patients, but to submit them to a protocol of frequent observation in order to avoid the morbidities associated with radical surgery. However, assessment of treatment response remains a significant challenge due to the subjectivity of the clinical examination and to the lack of sufficiently sensitive tools to ensure the absence of tumor cells or to detect early disease recurrence. Circulating DNA carrying tumor-specific genetic alterations (circulating tumor DNA - ctDNA) can be found in the cell-free fraction of the blood and has been successfully used to monitor the tumor dynamics in solid tumors. Recent advances in sequencing technologies have enabled the rapid and cost effective identification of genetic alterations in individual tumors, overcoming the problem imposed by the absence of recurrent genetic alterations in these tumors. These alterations can be used as personalized biomarkers to monitor treatment response, detect residual disease and early tumor recurrence. The purpose of this work was to identify and validate the use of personalized biomarkers for patients with locally advanced rectal cancer treated with nCRT and to evaluate the ability of these biomarkers to monitor the tumor dynamics, to define surgical approach and to detect early recurrence of the disease. Pre-treatment biopsies from 6 patients with cT2-3N0-1M0 distal rectal adenocarcinoma were prospectively collected. Genomic DNA extracted from the biopsies was used to construct mate-pair libraries for whole genome sequencing using SOLiD platform. Inter and intrachromosomal rearrangements were identified using an in-house bioinformatics pipeline and validated using PCR amplification and Sanger sequencing. At least three structural variations were validated for each patient. Plasma samples were collect at diagnosis, after nCRT and follow-up. Circulating DNA was obtained from the plasma samples and personalized assays were designed to monitor the presence of structural variations using Droplet Digital PCR. ctDNA was detected in all pre-treatment plasma samples for patients with T3 tumors. The detection of these biomarkers showed a good correlation with the treatment response, nonetheless, the approach was not sensitive enough to detect residual disease. In two patients who developed metastatic disease, an increase in ctDNA levels was observed at least 36 weeks before clinical detection of metastatic disease, and it was possible to correlate the level of ctDNA in subsequent plasma samples with response to the second-line treatment. This study, although exploratory, generated relevant and sufficient data to support additional studies to evaluate the use of personalized biomarkers in the surgical management and follow-up of rectal cancer patients treated with nCRT


Assuntos
Masculino , Feminino , Biomarcadores Tumorais/análise , DNA/genética , Terapia Neoadjuvante/classificação , Neoplasias Retais/patologia , Biblioteca Gênica , Células Neoplásicas Circulantes/metabolismo , Reação em Cadeia da Polimerase/métodos
5.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-89401

RESUMO

PURPOSE: To describe clinical findings in a Korean family with Axenfeld-Rieger syndrome. METHODS: A retrospective review of clinical data about patients with diagnosed Axenfeld-Rieger syndrome. Five affected members of the family underwent a complete ophthalmologic examination. We screened the forkhead box C1 gene and the pituitary homeobox 2 gene in patients. Peripheral blood leukocytes and buccal mucosal epithelial cells were obtained from seven members of a family with Axenfeld-Rieger syndrome. DNA was extracted and amplified by polymerase chain reaction, followed by direct sequencing. RESULTS: The affected members showed iris hypoplasia, iridocorneal adhesions, posterior embryotoxon, and advanced glaucoma in three generation. None had systemic anomalies. Two mutations including c.1362_1364insCGG and c.1142_1144insGGC were identified in forkhead box C1 in four affected family members. CONCLUSIONS: This study may help to understand clinical findings and prognosis for patients with Axenfeld-Rieger syndrome.


Assuntos
Idoso de 80 Anos ou mais , Segmento Anterior do Olho/anormalidades , DNA/genética , Análise Mutacional de DNA , Anormalidades do Olho/diagnóstico , Feminino , Fatores de Transcrição Forkhead/genética , Testes Genéticos , Proteínas de Homeodomínio/genética , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Linhagem , Estudos Retrospectivos , Fatores de Transcrição/genética , Adulto Jovem
6.
Indian J Exp Biol ; 2014 Oct; 52(10): 1011-1016
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-153801

RESUMO

DNA from molted feathers is being increasingly used for genetic studies on birds. However, the DNA obtained from such non-invasive sources is often not of enough quantity and quality for isolation of new microsatellite markers. The present study examined the potential of shed feathers of near threatened Painted Stork as a source of its DNA for cross-species amplification of microsatellites. Thirty-one shed feathers of varying conditions (‘good’ and ‘deteriorated’) and sizes (‘large’, ‘intermediate’ and ‘small’) collected in a north Indian population were used to isolate DNA by a standard isopropanol method and 11 microsatellite markers already developed in the Wood Stork were screened for amplification. Nine plucked feathers from two dead Painted Storks were also used to compare the DNA yield and amplification success. The DNA yield of feathers varied significantly in relation to the calamus size and condition. Among molted feathers, ‘good’ and ‘large’ samples provided more DNA than ‘deteriorated’ and ‘small’ ones, respectively. ‘Large’ plucked feathers yielded more DNA than ‘large’ molted feathers. DNA was almost degraded in all the samples and ratio of absorbance at 260/280 nm varied from 1.0 to 1.8, indicating impurity in many samples. Independent of DNA yields, all microsatellites were cross-amplified in all kinds of feathers, with >80% success in different feather categories. It is concluded that the shed feathers can be successfully used to isolate DNA in the Painted Stork and for cross-species amplification of microsatellites.


Assuntos
Animais , Aves/genética , DNA/genética , Plumas/química , Genética Populacional/métodos , Repetições de Microssatélites , Especificidade da Espécie
7.
Indian J Biochem Biophys ; 2014 Apr; 51(2): 115-120
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-154247

RESUMO

White matter disease refers to a set of diseases that affect the white matter of the brain and all of which have different consequences on brain function. Most of the studies have shown that it results from the defects during protein synthesis, with the gene defects in EIF2B1–5, encoding the five subunits of eukaryotic translation initiation factor 2B (eIF2B) α, β, γ, δ and ε, respectively. eIF2B plays a crucial role in protein translation and its regulation under different conditions. The previous studies have shown that mutations in five subunits of eIF2B cause white matter disease of the brain and thus EIF2B is the main culprit in development of white matter disease. In this study, the mutational screening of EIF2B5 gene encoding eIF2Bε was performed for the first time in 12 Kashmiri patients, each having a unique white matter disease condition. We found two novel missense mutations in EIF2B5: c.580A>G, p.Thr194Ala and c.611C>T, p.Ala204Val among the patients with demyelinating disease (multiple sclerosis), but no mutation was found in other patients. In conclusion our study suggests involvement of the EIF2B5 gene in MS development, thus suggesting p.Thr194Ala to be a susceptibility factor for the development of multiple sclerosis.


Assuntos
Estudos de Casos e Controles , DNA/sangue , DNA/genética , Fator de Iniciação 2B em Eucariotos/química , Fator de Iniciação 2B em Eucariotos/genética , Éxons/genética , Predisposição Genética para Doença , Humanos , Índia , Leucoencefalopatias/genética , Esclerose Múltipla/sangue , Esclerose Múltipla/genética , Mutação de Sentido Incorreto/genética , Conformação Proteica
8.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-143101

RESUMO

An 87-year-old woman visited our clinic for a scheduled cataract surgery. At the time of preoperative evaluation, slit lamp examination showed lattice-shaped and granular deposits with asymmetrical patterns in the stroma of both corneas. Genomic DNA samples of the patient, amplified by polymerase chain reaction, showed a single nucleotide substitution, c. 1580T>G (p.L527R), in the transforming growth factor-beta-induced TGFBI gene. We also found two additional SNP mutations, c.1620T>C (p.F540F) and c.1678+23G>A, along with the well-known L527R mutation. This is the first report of lattice corneal dystrophy type IV with an L527R mutation outside of Japan, and could challenge the idea that L527R is caused by a mutation from a single Japanese ancestor.


Assuntos
Idoso de 80 Anos ou mais , Distrofias Hereditárias da Córnea/diagnóstico , DNA/genética , Análise Mutacional de DNA , Proteínas da Matriz Extracelular/genética , Feminino , Humanos , Mutação , Linhagem , Reação em Cadeia da Polimerase , Fator de Crescimento Transformador beta/genética
9.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-143096

RESUMO

An 87-year-old woman visited our clinic for a scheduled cataract surgery. At the time of preoperative evaluation, slit lamp examination showed lattice-shaped and granular deposits with asymmetrical patterns in the stroma of both corneas. Genomic DNA samples of the patient, amplified by polymerase chain reaction, showed a single nucleotide substitution, c. 1580T>G (p.L527R), in the transforming growth factor-beta-induced TGFBI gene. We also found two additional SNP mutations, c.1620T>C (p.F540F) and c.1678+23G>A, along with the well-known L527R mutation. This is the first report of lattice corneal dystrophy type IV with an L527R mutation outside of Japan, and could challenge the idea that L527R is caused by a mutation from a single Japanese ancestor.


Assuntos
Idoso de 80 Anos ou mais , Distrofias Hereditárias da Córnea/diagnóstico , DNA/genética , Análise Mutacional de DNA , Proteínas da Matriz Extracelular/genética , Feminino , Humanos , Mutação , Linhagem , Reação em Cadeia da Polimerase , Fator de Crescimento Transformador beta/genética
10.
Indian J Exp Biol ; 2013 Dec; 51(12): 1130-1136
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-150302

RESUMO

The genomic and cDNA sequences of BnSUT1C were isolated from B. napus. Combination of cDNA and genomic DNA sequences revealed that the BnSUT1C gene contained three exons and two introns. The cDNA encodes a protein of 513 amino acids with a calculated molecular mass of 54.7 kDa and an isoelectric point of 9.12. It exhibits typical features of sucrose transporter with 12 trans-membranes spanning domains. BnSUT1C showed highly homologous with AtSUC1 and AtSUC5. A histidine residue, which is conserved across all functional sucrose transporter proteins in higher plants, is located at position 66 of the BnSUT1C. Two putative pollen-specific cis-elements, AGAAA and GTGA motifs, are located in 5′-upstream of BnSUT1C. The spatial and temporal expression patterns carried out by semi-quantitative RT-PCR and Real-Time PCR, which indicated that BnSUT1C predominantly expressed in later developmental stages of anther, as tapetal cells began to shrink and collapse. BnSUT1C could mediate the uptake of sucrose in the pollen and retrieval of tapetal degenerated products during pollen maturation.


Assuntos
Sequência de Aminoácidos , Arabidopsis/genética , Sequência de Bases , Brassica napus/genética , Clonagem Molecular , DNA/genética , DNA Complementar/genética , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/isolamento & purificação , Proteínas de Plantas/genética , Proteínas de Plantas/isolamento & purificação , RNA Mensageiro/genética , Homologia de Sequência de Aminoácidos , Transcriptoma
11.
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-159949

RESUMO

Background: Fine needle aspiration cytology (FNAC) is most commonly used in first line investigation for tuberculous lymphadenopathy (TBLAP). Real-time polymerase chain reaction (PCR) an extremely versatile technique is being used for diagnosis and follow up of patients with infectious diseases. It can also be used for detecting Mycobacterium tuberculosis (Mtb) DNA in FNAC samples of TBLAP for rapid diagnosis and treatment. Aim: Detection of Mtb DNA on FNAC samples of tuberculous lymphadenopathy using Real-time PCR. Material and Methods: Twelve clinico-cytologically diagnosed TBLAP cases and five controls were included in the study. FNA samples were used for studying morphology, AFB demonstration, culture and for detecting Mtb DNA using Real-time PCR. Results: Mtb DNA was detected in ten cases (83.33 %) by Real-time PCR. ZN stain was positive in eight cases and culture in six cases. Conclusion: Detection of Mtb DNA in FNAC samples using Real-time PCR is a time saving, logical, economical approach over the culture based method.


Assuntos
Adolescente , Adulto , Criança , DNA/análise , DNA/genética , Humanos , Pessoa de Meia-Idade , Mycobacterium/genética , Reação em Cadeia da Polimerase em Tempo Real/estatística & dados numéricos , Tuberculose dos Linfonodos/citologia , Tuberculose dos Linfonodos/genética
12.
Indian J Hum Genet ; 2013 Jan; 19(1): 78-83
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-147640

RESUMO

CONTEXT: Amplification of Guanine-Cytosine (GC) -rich sequences becomes important in screening and diagnosis of certain genetic diseases such as diseases arising due to expansion of GC-rich trinucleotide repeat regions. However, GC-rich sequences in the genome are refractory to standard polymerase chain reaction (PCR) amplification and require a special reaction conditions and/or modified PCR cycle parameters. AIM: Optimize a cost effective PCR assay to amplify the GC-rich DNA templates. SETTINGS AND DESIGN: Fragile X mental retardation gene (FMR 1) is an ideal candidate for PCR optimization as its GC content is more than 80%. Primers designed to amplify the GC rich 5’ untranslated region of the FMR 1 gene, was selected for the optimization of amplification using DNA extracted from buccal mucosal cells. MATERIALS AND METHODS: A simple and rapid protocol was used to extract DNA from buccal cells. PCR optimization was carried out using three methods, (a) substituting a substrate analog 7-deaza-dGTP to dGTP (b) in the presence of a single PCR additive and (c) using a combination of PCR additives. All PCR amplifications were carried out using a low-cost thermostable polymerase. RESULTS: Optimum PCR conditions were achieved when a combination of 1M betaine and 5% dimethyl sulfoxide (DMSO) was used. CONCLUSIONS: It was possible to amplify the GC rich region of FMR 1 gene with reproducibility in the presence of betaine and DMSO as additives without the use of commercially available kits for DNA extraction and the expensive thermostable polymerases.


Assuntos
Bochecha/citologia , Citosina/análogos & derivados , DNA/genética , Elementos Facilitadores Genéticos/genética , Síndrome do Cromossomo X Frágil/genética , Guanina/análogos & derivados , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase/métodos
13.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-19707

RESUMO

PURPOSE: To assess whether the expression of heat shock protein 72 (Hsp72) protects rat retinal ganglion cells (RGC-5) from apoptotic cell death. METHODS: Hsp72 expression in RGC-5 cells transduced with replication-deficient recombinant adenovirus was analyzed by Western blot analysis and immunofluorescence. The effect of Hsp72 expression on etoposide-induced apoptotic cell death was examined by microscopic analysis and confirmed by cell proliferation assay. RESULTS: Western blot analysis and immunofluorescence clearly showed adenovirus-mediated Hsp72 expression in RGC-5 cells. Treatment with etoposide resulted in the death of a proportion of the cells by apoptosis. However, this apoptotic cell death was significantly reduced in cells expressing Hsp72, with the reduction in cell death correlating to the level of Hsp72 expression. CONCLUSIONS: Over-expression of Hsp72 alone is sufficient to rescue neuronal cells from apoptotic cell death, suggesting that fine-tuning its expression may be an effective neuroprotective approach in retinal degenerative disease.


Assuntos
Animais , Western Blotting , Morte Celular/genética , Sobrevivência Celular , Células Cultivadas , DNA/genética , Modelos Animais de Doenças , Etoposídeo/toxicidade , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP72/biossíntese , Imuno-Histoquímica , Ratos , Degeneração Retiniana/genética , Células Ganglionares da Retina/efeitos dos fármacos
14.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-205012

RESUMO

To report a novel mutation within the CHST6 gene, as well as describe light and electron microscopic features of a case of macular corneal dystrophy. A 59-year old woman with macular corneal dystrophy in both eyes who had decreased visual acuity underwent penetrating keratoplasty. Further studies including light and electron microscopy, as well as DNA analysis were performed. Light microscopy of the cornea revealed glycosaminoglycan deposits in the keratocytes and endothelial cells, as well as extracellularly within the stroma. All samples stained positively with alcian blue, colloidal iron, and periodic acid-Schiff. Electron microscopy showed keratocytes distended by membrane-bound intracytoplasmic vacuoles containing electron-dense fibrillogranular material. These vacuoles were present in the endothelial cells and between stromal lamellae. Some of the vacuoles contained dense osmophilic whorls. A novel homozygous mutation (c.613 C>T [p.Arg205Trp]) was identified within the whole coding region of CHST6. A novel CHST6 mutation was detected in a Korean macular corneal dystrophy patient.


Assuntos
Distrofias Hereditárias da Córnea/diagnóstico , Ceratócitos da Córnea/ultraestrutura , DNA/genética , Análise Mutacional de DNA , Feminino , Humanos , Microscopia Eletrônica , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Linhagem , Reação em Cadeia da Polimerase , República da Coreia , Sulfotransferases/genética
15.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-174792

RESUMO

Bovine spongiform encephalopathy (BSE) is one of the fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs) caused by infectious prion proteins. Genetic variations correlated with susceptibility or resistance to TSE in humans and sheep have not been reported for bovine strains including those from Holstein, Jersey, and Japanese Black cattle. Here, we investigated bovine prion protein gene (PRNP) variations in Hanwoo cattle [Bos (B.) taurus coreanae], a native breed in Korea. We identified mutations and polymorphisms in the coding region of PRNP, determined their frequency, and evaluated their significance. We identified four synonymous polymorphisms and two non-synonymous mutations in PRNP, but found no novel polymorphisms. The sequence and number of octapeptide repeats were completely conserved, and the haplotype frequency of the coding region was similar to that of other B. taurus strains. When we examined the 23-bp and 12-bp insertion/deletion (indel) polymorphisms in the non-coding region of PRNP, Hanwoo cattle had a lower deletion allele and 23-bp del/12-bp del haplotype frequency than healthy and BSE-affected animals of other strains. Thus, Hanwoo are seemingly less susceptible to BSE than other strains due to the 23-bp and 12-bp indel polymorphisms.


Assuntos
Animais , Sequência de Bases , Bovinos , DNA/genética , Encefalopatia Espongiforme Bovina/genética , Variação Genética , Haplótipos , Príons/genética , República da Coreia
16.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-214940

RESUMO

PURPOSE: The purpose of this study was to determine the pharmacogenetic effects of complement factor H (CFH) Y402H, LOC387715 and high-temperature requirement factor A1 (HTRA1) genotypes on the treatment of exudative age-related macular degeneration (AMD) by intravitreal bevacizumab injection in a Korean population. METHODS: Seventy-five patients diagnosed with exudative AMD were treated with intravitreal bevacizumab (2.5 mg) monotherapy. All patients received three initial intravitreal bevacizumab injections every four weeks and were then treated "as needed" based on clinical findings, optical coherence tomography and fluorescein angiography during the 12 month follow-up period after the third injection. RESULTS: The difference in visual acuity improvement among the three genotypes of LOC387715 were statistically significant at six months post-treatment (logarithm of the minimum angle of resolution; TT, 0.346; GT, 0.264; GG, 0.188; p = 0.037). Among the LOC387715 genotypes, the number of additional injections was lower in patients who had the risk T allele (GG, 2.143; GT, 2.000; TT, 1.575; p = 0.064). There was no significant difference between visual acuity and central macular thickness change in the CFH Y402H polymorphism group during the 12 month follow-up period. However, the TC group of CFH Y402H required more additional bevacizumab injections than the TT group (TT, 1.517; TC, 3.363; p = 0.020). CONCLUSIONS: This study demonstrated that different LOC387715/HTRA1 genotypes resulted in different bevacizumab treatment responses on exudative AMD. Patients with the risk allele had an improved treatment response and less need for additional injections. However, patients with the CFH Y402H risk allele needed more additional injections of bevacizumab in order to improve visual acuity. This study illustrates how pharmacogenetic factors may help determine treatment modality and dosing. This could ultimately provide basic data for 'personalized medicine' in AMD.


Assuntos
Idoso , Alelos , Inibidores da Angiogênese/administração & dosagem , Anticorpos Monoclonais Humanizados/administração & dosagem , DNA/genética , Feminino , Seguimentos , Genótipo , Humanos , Injeções Intravítreas , Degeneração Macular/tratamento farmacológico , Masculino , Farmacogenética/métodos , Polimorfismo Genético , Estudos Retrospectivos , Serina Endopeptidases/genética , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Acuidade Visual
17.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-214939

RESUMO

PURPOSE: To investigate whether the G6721T polymorphism (rs.7003908) of the non-homologous end-joining DNA repair XRCC7 gene contributes to the development of exudative age-related macular degeneration (ARMD). METHODS: The present case-control study consisted of 111 patients with exudative ARMD and 112 sex frequency-matched healthy controls that were randomly selected from unrelated volunteers in the same clinic. Genotypes were determined by the Restriction Fragment Length Polymorphism (PCR-RFLP) based method. Logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for ARMD risk associated with polymorphism of XRCC7. In all analysis the GG genotype was considered to be the reference genotype. RESULTS: There was no significant association between genotypes of XRCC7 and susceptibility to ARMD. Considering the significant difference in age distribution between cases and controls, age was used as a covariate in further analysis. After ORs were adjusted for age, the same result was observed. In the next step we stratified our subjects into outdoor and indoor groups according to their job titles. The outdoor and indoor patients were occupationally exposed to sunlight and not exposed to sunlight, respectively. Our present study showed that among indoor subjects there was no association between XRCC7 polymorphism and susceptibility to ARMD. However, among outdoor subjects, the GT + TT genotypes compared to the GG genotype increased the risk of ARMD (OR, 3.13; 95% CI, 1.04-9.39; p = 0.042). CONCLUSIONS: Our study revealed that the T allele of the G6721T polymorphism of XRCC7 increased the risk of ARMD among outdoor subjects.


Assuntos
Idoso , DNA/genética , Proteína Quinase Ativada por DNA/genética , Exposição Ambiental , Exsudatos e Transudatos , Feminino , Seguimentos , Predisposição Genética para Doença , Genótipo , Humanos , Degeneração Macular/genética , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares/genética , Reação em Cadeia da Polimerase , Polimorfismo Genético , Estudos Retrospectivos , Fatores de Risco
18.
Indian J Cancer ; 2011 Apr-Jun; 48(2): 223-229
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-144457

RESUMO

Aims : The aim of the present study is to investigate the association of polymorphism in cytochrome P450 2C9 (CYP2C9) with head and neck squamous cell carcinoma (HNSCC) and response in patients receiving chemoradiotherapy. Materials and Methods : One hundred ten males suffering from locally advanced head and neck squamous cell carcinoma and an equal number of healthy controls were genotyped for CYP2C9*2 and CYP2C9*3, leading to poor metabolizers (PMs) by PCR-based RFLP. Each case was assessed thoroughly for treatment response following WHO criteria. Results : The frequency of heterozygous genotypes of both CYP2C9*2 (27.3%) and CYP2C9*3 (20.1%) were found to be significantly higher in the HNSCC cases as compared to the healthy controls. Tobacco intake in the form of chewing or smoking and alcohol intake resulted in several fold increase in the risk to HNSCC in the cases carrying variant genotypes of CYP2C9*2 or CYP2C9*3. Further, majority of the cases assessed for response (134) carrying variant alleles of both CYP2C9*2 (65.3%) or CYP2C9*3 (70.58%) were found to respond poorly to the radio-chemotherapy. Conclusions : The data suggests a significant association of the CYP2C9 polymorphism with HNSCC and treatment outcome underlining the importance of pretherapeutic genotyping in determining the treatment schedule.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Hidrocarboneto de Aril Hidroxilases/genética , Braquiterapia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/terapia , Estudos de Casos e Controles , Quimiorradioterapia , DNA/genética , Seguimentos , Genótipo , Neoplasias de Cabeça e Pescoço/genética , Neoplasias de Cabeça e Pescoço/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Fumar , Taxa de Sobrevida , Resultado do Tratamento
19.
Artigo em Inglês | IMSEAR (Sudeste Asiático), GHL | ID: sea-135663

RESUMO

Background & objectives: Cyclin D1 has been strongly implicated in cell proliferation particularly in the G1/S checkpoint of the cell cycle, and prognoses in human malignancies. We investigated the correlation between cyclin D1 overexpression and clinicopathological features as well as cell cycle parameters to understand its clinical significance in patients with tobacco-related oral squamous cell carcinoma (OSCC). Methods: Immunohistochemistry for cyclin D1 and DNA flowcytometry for cell cycle parameters was done on paraffin embedded tumour samples from 45 patients with OSCC Results: Higher expression of cyclin D1 was observed only in 30 (66.6%) of 45 cases that correlated with advanced age (P <0.02), higher tumour stage ((P<0.01), histological differentiation and lymph node metastasis (P <0.01). Analysis of nuclear DNA pattern revealed cyclin D1 immunoreactivity in tumours with aggressive DNA pattern such as aneuploidy ((P<0.05) and higher S phase fraction ((P<0.04). Interpretation & conclusions: Higher expression of cyclin D1 in oral cancer appears to be closely linked to cell proliferation, differentiation and lymph node invasion. Pre-operative evaluation of cyclin D1 in biopsy specimen may be useful in planning the most appropriate treatment strategies in patients with tobacco-related OSCC.


Assuntos
Adulto , Idoso , Aneuploidia , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/secundário , Ciclo Celular , Ciclina D1/análise , Ciclina D1/biossíntese , Ciclina D1/genética , DNA/genética , Diploide , Feminino , Humanos , Imuno-Histoquímica , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/química , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos , Tabaco sem Fumaça/efeitos adversos
20.
Journal of Guilan University of Medical Sciences. 2011; 19 (76): 15-21
em Fa | IMEMR (Mediterrâneo Oriental) | ID: emr-110044

RESUMO

Various frequencies of the mtDNA mutations have been reported from different population world wild. Three mitochondrial DNA [mtDNA] mutations including A1555G, A 3243G, and A7445G which occurred in MTRNR1, MTTL1 and MTTS1 genes were considered as the main causes of mitochondrial hearing loss in some populations. To determine the frequency of the A1555G, A3243G, and A7445G mutations in nonsyndromic sensorineural hearing loss subjects in Gilan. Forty six subjects with nonsyndromic sensorineural hearing loss were screened by provided questionnaire and audiogram from Gillan Welfare Organization. PCR-RFLP procedure was used in order to presence the MtDNA of A1555GA 3243G and A7445G mutations and was confirmed by subsequent direct sequencing. There was no MtDNA of A1555G, A3243G and A7445G mutation in the cohort study of 46 deaf individuals. Investigation of PCR-RFLP of the MTTL1 gene for existence A3243G mutation lead to identification a G3316A variant that destroyed other restriction site, in the other site of PCR fragment. Our finding indicated that possibility the association of mitochondrial mutations with deafness is very low in deaf subjects in north of Iran. According to existence the G3316A that its pathogenesis in relation to hearing loss phenotype has not stabilized, the frequency of G3316A is 1.46% that can be had highlights role of mitochondrial mutation in deafness


Assuntos
Humanos , Surdez/genética , Mitocôndrias/genética , Predisposição Genética para Doença , Análise Mutacional de DNA , Programas de Rastreamento , Inquéritos e Questionários , Testes Auditivos , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , DNA/genética , Fenótipo
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