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1.
Rev. int. androl. (Internet) ; 18(1): 7-13, ene.-mar. 2020. tab, graf
Artigo em Inglês | IBECS | ID: ibc-193832

RESUMO

OBJECTIVES: The aim of this study was to evaluate polymorphisms of sperm protamine genes and their effects on the result of CMA3 staining in varicocele men. MATERIAL AND METHODS: In a case control study, 128 patients with male infertility due to varicocele and 128 controls were recruited. Polymorphisms of PRM1 and PRM2 genes in extracted DNA samples were assessed by PCR-SSCP and sequencing. Protamine deficiency was also indirectly estimated by CMA3 staining. RESULT: Nine different variants including six variants in PRM1 gene and three variants in PRM2 gene were found among varicocele patients. The results showed that sperm count, motility and morphology were significantly different between control group without gene variations and varicocele group who had several variations in their protamine genes (P<0.05). CONCLUSION: Therefore, PRM1 and PRM2 variations in varicocele patients are associated with the production of spermatozoa with more protamine deficiency and this is one of the possible causes of infertility due to varicocele


OBJETIVOS: El objetivo de este estudio fue evaluar los polimorfismos de los genes de la protamina espermática y sus efectos sobre el resultado de la tinción CMA3 en varones con varicocele. MATERIAL Y MÉTODOS: En un estudio de casos y controles, se reclutó a 128 varones con infertilidad por varicocele y 128 controles. Los polimorfismos de los genes PRM1 y PRM2 en muestras de ADN extraídas se evaluaron mediante PCR-SSCP y se realizó su secuenciación. La deficiencia de protamina también se estimó indirectamente mediante tinción CMA3. RESULTADO: Se encontraron 9 variantes diferentes, incluidas 6 variantes en el gen PRM1 y 3 variantes en el gen PRM2 entre los pacientes con varicocele. Los resultados mostraron que el recuento de espermatozoides, la movilidad y la morfología eran considerablemente diferentes entre el grupo de control sin variaciones genéticas y el grupo de varicocele que presentaba algunas variaciones en sus genes de protamina (p < 0,05). CONCLUSIÓN: Por tanto, las variaciones de PRM1 y PRM2 en los pacientes con varicocele están asociadas con la producción de espermatozoides con más deficiencia de protamina y esta es una de las posibles causas de infertilidad debida al varicocele


Assuntos
Humanos , Masculino , Adulto , Varicocele/complicações , Varicocele/genética , Polimorfismo Genético , Infertilidade Masculina/etiologia , Infertilidade Masculina/genética , Protaminas/genética , Estudos de Casos e Controles , Polimorfismo Conformacional de Fita Simples , Coloração e Rotulagem
2.
Trop Anim Health Prod ; 52(4): 2091-2099, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32052250

RESUMO

Goat production under migratory system is foremost meat resource in Western Himalayan region of India. Thus, selection of goats for superior growth rate is rewarding. Growth hormone (GH) gene is identified as main regulator of post-natal growth and development. The objective of this study was to identify GH gene variants in Gaddi goats reared under migratory system via polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single-strand conformation polymorphism (SSCP). Blood samples from 63 animals from different migratory flocks registered under All India Coordinated Research Project, Himachal Pradesh Agricultural University (HPAU), Palampur, were subjected to DNA isolation. A total of 422, 116, 389 and 181-bp amplicons were generated on amplification of four targeted regions of GH gene. GH1 and GH2 fragments were analysed using PCR-RFLP (HaeIII RE) that revealed three variants (AA, AB and BB) for GH1 having frequency as 0.27, 0.52 and 0.31, respectively whereas, two variants (AB and BB) were revealed for GH2 fragment having frequency of 0.24 and 076, respectively. GH3 and GH4 fragments were subjected to PCR-SSCP that detected three genotypes (AB, BB and AA) for GH3 with respective genotype frequency as 0.57, 0.21 and 0.22 respectively; however, GH4 was found to be monomorphic. The polymorphism information content values for GH1, GH2 and GH3 were 0.37, 0.36 and 0.34, respectively, which suggested the median level of polymorphism at studied loci and also indicated the effectiveness of the studied marker for population genetic studies. Significant associations (P ≤ 0.05) were detected for GH1 with 9-month body weight, GH2 with 9 and 12-month heart girth and GH3 with 6-month body weight, body height and body length, respectively. From the present study, it was concluded that SNPs and their association with some body measurements may be employed as useful markers for ongoing phenotypic selection programme.


Assuntos
Criação de Animais Domésticos , Cabras/genética , Hormônio do Crescimento/genética , Polimorfismo de Nucleotídeo Único , Animais , Frequência do Gene , Genótipo , Cabras/crescimento & desenvolvimento , Índia , Fenótipo , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único/fisiologia , Polimorfismo Conformacional de Fita Simples
3.
Gene ; 740: 144400, 2020 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-31987910

RESUMO

Myostatin (MSTN) is a circulating factor that is secreted by muscle cells, and that acts upon those cells to inhibit the proliferation of muscle fibres during pre-natal muscle growth. The Polymerase Chain Reaction (PCR) coupled with Single Strand Conformational Polymorphism (SSCP) analysis, was used to reveal variation in the bovine MSTN gene (MSTN) in 722 cattle from a variety of breeds farmed in New Zealand (NZ). These included Hereford, Angus, Charolais, Simmental, Red Poll, South Devon, Shorthorn, Murray Grey, cross-bred Holstein-Friesian × Jersey cattle, and other composite breeds of cattle. Sequence analysis of five regions of MSTN that encompassed coding and non-coding regions of the gene, revealed a total of twelve single-nucleotide substitutions (7 in intron 1 and 5 in a region spanning the intron 2 - exon 3 boundary), and a single nucleotide deletion. Of these 12 substitutions, five are reported here for the first time, whereas seven have been previously described. The deletion c.748-78del, was located in the intron 2 - exon 3 boundary region, and has been reported previously. No nucleotide variation was identified in exons 1, 2 and 3. A total of 18 extended haplotypes were resolved spanning two variable regions (intron 1 and the intron 2 - exon 3 boundary), some of which were common across the breeds, while others were peculiar to particular breeds. The genetic variations identified provide insight into the conserved and polymorphic nature of the coding and non-coding sequences of bovine MSTN respectively, and thus provides a baseline for further study into how variation in the gene might affect growth and carcass traits in NZ cattle.


Assuntos
Bovinos/genética , Variação Genética , Miostatina/genética , Animais , Nova Zelândia , Polimorfismo Conformacional de Fita Simples
4.
Methods Mol Biol ; 2102: 373-394, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31989568

RESUMO

Sputum and plasma can provide noninvasive materials to investigate biomarkers for cancer detection and diagnosis. Mutations in the K-ras oncogene and p53 suppressor gene have been frequently found in sputum and plasma samples collected not only from lung cancer patients but also in those of patients prior to presenting clinical symptoms of lung cancer, suggesting that they may also provide useful biomarkers from early lung cancer diagnosis. However, the detection of these mutations has been complicated by the fact that they often occur in only a small fraction of epithelial cells among sputum cells, and of cell-free DNA present in plasma. This chapter describes methods to isolate low fraction epithelial cells from sputum and cell-free DNA from plasma samples obtained from lung cancer patients and to identify low fraction K-ras and p53 mutations in these samples.


Assuntos
DNA Tumoral Circulante/sangue , Análise Mutacional de DNA/métodos , DNA de Neoplasias/genética , Genes p53/genética , Neoplasias Pulmonares/genética , Proteínas Proto-Oncogênicas p21(ras)/genética , Escarro/química , DNA de Neoplasias/análise , DNA de Neoplasias/química , DNA de Neoplasias/isolamento & purificação , Células Epiteliais/química , Genes ras , Humanos , Mutação , Mutação Puntual , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Escarro/citologia , Fluxo de Trabalho
5.
Exp Appl Acarol ; 80(2): 247-256, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31953633

RESUMO

PCR-based single-strand conformation polymorphism (SSCP) analyses combined with DNA sequencing of the prokaryotic 16S ribosomal (r) RNA gene encompassing the hypervariable V4 region was used to determine the bacterial composition of Rocky Mountain wood ticks (Dermacentor andersoni) attached to Richardson's ground squirrels (Urocitellus richardsonii) and questing on vegetation in southern Saskatchewan, Canada. The bacteria present in questing adult ticks from Saskatchewan Landing Provincial Park included Rickettsia peacockii, a Francisella-like endosymbiont (FLE) and an Arsenophonus-like endosymbiont. Bacteria in the adult and nymphal ticks attached to U. richardsonii collected from Beechy included R. peacockii, a FLE, and several other genera (e.g., Ralstonia, Sphingobium, Comamonas and Pseudomonas). The bacteria detected in D. andersoni in the present study are consistent with the findings of other studies that have characterized the microbiome of this tick species in the USA using next generation sequencing. This result demonstrates that the SSCP-based approach used in this study is cost- and time-effective for examining bacterial composition in ticks.


Assuntos
Dermacentor/microbiologia , Polimorfismo Conformacional de Fita Simples , Rickettsia/classificação , Animais , Canadá , DNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
6.
Immunogenetics ; 71(10): 647-663, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31761978

RESUMO

The classical class I and class II molecules of the major histocompatibility complex (MHC) play crucial roles in immune responses to infectious pathogens and vaccines as well as being important for autoimmunity, allergy, cancer and reproduction. These classical MHC genes are the most polymorphic known, with roughly 10,000 alleles in humans. In chickens, the MHC (also known as the BF-BL region) determines decisive resistance and susceptibility to infectious pathogens, but relatively few MHC alleles and haplotypes have been described in any detail. We describe a typing protocol for classical chicken class I (BF) and class II B (BLB) genes based on a hybridization method called reference strand-mediated conformational analysis (RSCA). We optimize the various steps, validate the analysis using well-characterized chicken MHC haplotypes, apply the system to type some experimental lines and discover a new chicken class I allele. This work establishes a basis for typing the MHC genes of chickens worldwide and provides an opportunity to correlate with microsatellite and with single nucleotide polymorphism (SNP) typing for approaches involving imputation.


Assuntos
Genes MHC da Classe II/genética , Genes MHC Classe I/genética , Hibridização de Ácido Nucleico/métodos , Polimorfismo Genético , Análise de Sequência de DNA/normas , Animais , Galinhas , Polimorfismo Conformacional de Fita Simples , Padrões de Referência , Análise de Sequência de DNA/métodos
9.
Growth Factors ; 37(3-4): 153-163, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31500477

RESUMO

The objective of this study was to test the association of the variation in a 360 bp region in exon 2 of the ovine bone morphogenetic protein 4 (BMP4) gene with growth performance (birth weight, pre-weaning average daily gain, weaning weight, post-weaning average daily gain and marketing weight) and body conformational traits (height at withers, height at hips, body length, heart girth, thigh circumference, body mass index, skeletal muscle index, body index and relative body index) in 242 Barki lambs using polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP). Two variants (A and B) and three genotypes (AA, AB and BB) were detected. The BMP4 genotype significantly affected (p < .05 or p < .01) post-weaning daily gain, marketing weight, height at hips, thigh circumference, body mass index and skeletal muscle index. The results provided valuable information indicating selection for the BMP4 genotype might increase growth and muscularity in Barki lambs.


Assuntos
Composição Corporal/genética , Tamanho Corporal/genética , Proteína Morfogenética Óssea 4/genética , Ovinos/crescimento & desenvolvimento , Ovinos/genética , Animais , Sequência de Bases , Éxons/genética , Feminino , Variação Genética/genética , Genótipo , Masculino , Fenótipo , Polimorfismo Conformacional de Fita Simples/genética , Análise de Sequência de DNA , Ganho de Peso/genética
11.
Methods Mol Biol ; 2015: 79-104, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31222698

RESUMO

Single-strand conformation polymorphism (SSCP) is a popular method used to study the genetic heterogeneity and population variability of Citrus tristeza virus (CTV) isolates. It is a simple, low-cost, and highly specific method for mutation detection of specific genes, mostly of the CTV major coat protein gene (p25). The technique is based on a comparison on polyacrylamide gel of electrophoretic profiles of single-stranded (ss) DNA sequences in terms of their spatial conformation. SSCP involves cDNA synthesis and amplification of the target gene, denaturation of single strands, and electrophoresis in non-denaturing conditions. The ssDNAs can be afterward visualized by staining the polyacrylamide gel. Alternatively, using fluorescently labeled primers, the procedure can be performed in automated sequencers equipped with an appropriate capillary (CE-SSCP), which increases the potential of high-throughput analysis, precision, and the reproducibility of results. CE-SSCP can be also directly applied to the virus particles obtained by elution from ELISA plates or tissue-print membranes.


Assuntos
Closterovirus/genética , Polimorfismo Conformacional de Fita Simples/genética , Eletroforese em Gel de Poliacrilamida , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes
12.
Poult Sci ; 98(10): 4327-4337, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31111951

RESUMO

Consumer preference for slow-growing broiler chickens is rising because of increased demand for high-quality poultry products. Korat chicken (KRC) is a slow-growing chicken generated in Thailand. A goal of the KRC breeding program is to produce meat with a low purine content to benefit an aging population, without interfering with growth performance. Thus, this study aimed to investigate the effects of genes encoding melanocortin 4 receptor (MC4R), calpain 1 (CAPN1), and adenylosuccinate lyase (ADSL) on body weight, muscle fiber, and content of purine and its derivatives (i.e., adenine, guanine, hypoxanthine, and xanthine), to develop molecular markers for breeding programs. Genotypes of MC4R, CAPN1, and ADSL were obtained from 583 KRCs by PCR-single-strand conformation polymorphism. The body weight and purine contents of the KRCs were measured every 2 wk until the KRCs reached market weight at 10 wk of age. A significant association between the MC4R genotype and body weight at 2, 4, and 10 wk of age was detected. KRC possessing the BB genotype of CAPN1 showed significantly heavier body weight at 6 wk of age and guanine content at 4 wk of age, and a smaller muscle fiber diameter in the breast muscle at 10 wk of age, compared with those of the other genotypes. In addition, high expression levels of the CAPN1 and ADSL genes were detected in the breast muscle at 2 wk of age. Although higher purine contents were detected at a young age, no significant associations with the MC4R, CAPN1, and ADSL genes were detected. Our results indicate that MC4R and CAPN1 could be used as genetic markers for growth and meat quality in the slow-growing chicken breeding program.


Assuntos
Proteínas Aviárias/genética , Peso Corporal/genética , Galinhas/fisiologia , Purinas/metabolismo , Adenilossuccinato Liase/genética , Adenilossuccinato Liase/metabolismo , Animais , Proteínas Aviárias/metabolismo , Calpaína/genética , Calpaína/metabolismo , Galinhas/genética , Galinhas/crescimento & desenvolvimento , Feminino , Marcadores Genéticos , Genótipo , Masculino , Carne/análise , Polimorfismo Conformacional de Fita Simples , Receptor Tipo 4 de Melanocortina/genética , Receptor Tipo 4 de Melanocortina/metabolismo
13.
Asian Pac J Cancer Prev ; 20(5): 1339-1343, 2019 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-31127887

RESUMO

Background: The use of targeted specific genes in therapeutic and treatment decisions has been considered for lung cancer. The epidermal growth factor receptor (EGFR) gene, which is over expressed in non-small cell lung cancer (NSCLC), was considered as one of the targeted specific genes. EGFR mutations in exons 18­21, which encode a portion of the EGFR kinase domain, were found in NSCLC patients and were associated with the response of EGFRtyrosine kinase inhibitors (EGFR-TKIs). Therefore, a molecular technique for EGFR mutation detection has important benefits for therapy in NSCLC patients. This study aims to determine the EGFR mutations in patients with NSCLC using polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) in exons 18-21. Methods: DNA samples were extracted from formalin fixed paraffin embedded tissues of NSCLC patients who attended hospital. The extracted DNA was used as a template for the EGFR gene amplification. Results: Occurrence of EGFR mutations were found in 29 out of 50 cases (58%).The frequency of EGFR mutations by first PCR at exon 18, 19, 20 and 21 were 6 (12%), 19 (38%) 20 (40%) and at 21 (42%), respectively. By PCR-SSCP, the frequencies of EGFR mutations at exon 18, 19, 20 and 21 were 3(6%), 18(36%), 23(46%) and 13(26%), respectively. All of the mutations found were in agreement with DNA sequencings. Conclusion: The high frequency of EGFR mutations in NSCLC suggests that PCR-SSCP is a efficient screening method and useful for treatment plan.


Assuntos
Biomarcadores Tumorais/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/genética , Mutação , Reação em Cadeia da Polimerase/métodos , Polimorfismo Conformacional de Fita Simples , Adenocarcinoma/epidemiologia , Adenocarcinoma/genética , Adenocarcinoma/patologia , Adenocarcinoma Bronquioloalveolar/epidemiologia , Adenocarcinoma Bronquioloalveolar/genética , Adenocarcinoma Bronquioloalveolar/patologia , Adulto , Idoso , Sequência de Bases , Carcinoma Pulmonar de Células não Pequenas/epidemiologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma de Células Escamosas/epidemiologia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Estudos de Casos e Controles , Receptores ErbB/genética , Feminino , Seguimentos , Formaldeído/química , Humanos , Neoplasias Pulmonares/epidemiologia , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Inclusão em Parafina/métodos , Prognóstico , Homologia de Sequência , Tailândia/epidemiologia
14.
Gene ; 701: 131-138, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30905811

RESUMO

MicroRNAs (miRNAs) play an important role in animal growth and disease development, and sequence variation in microRNAs can alter their functions. Herein, we explored the effects of mutations in the miRNA-215 precursor sequence on the miRNA-215 regulatory network and resistance to Escherichia coli (E. coli). Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to detect sequence variations in Sutai and Meishan pigs. The miR-192 precursor sequence was not mutated, but the miR-215 precursor included an AT insertion mutation at position 6 (start from the first base of the miR-215 precursor) and a C/T mutation at position 43. Wild-type (WT) and mutant miR-215 precursor expression vectors were constructed to investigate the effects of sequence variation on expression of miR-215 and its target genes DLG5 and ALCAM, cytokine levels and E. coli adhesion. Compared with the WT control group, cells harbouring the C/T mutant vector displayed reduced miR-215 expression, increased target gene expression, elevated cytokine levels and rising E. coli adhesion, whereas cells harbouring the AT insertion mutant vector were not significantly changed. The sequence variation in the miRNA-215 precursor may affect the miRNA-215 regulatory network, and alter the stability of intestinal epithelial cells (IPEC-J2 cells) and resistance to E. coli. Our findings provide guidance for future research on the regulatory mechanisms of miR-215 in porcine resistance to E. coli F18, and identifying effective genetic markers against this organism.


Assuntos
Resistência à Doença/genética , Infecções por Escherichia coli/genética , Escherichia coli , MicroRNAs/genética , Mutagênese Insercional , Polimorfismo Conformacional de Fita Simples , Doenças dos Suínos/genética , Suínos/genética , Animais , Linhagem Celular , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , MicroRNAs/imunologia , Reação em Cadeia da Polimerase , Suínos/imunologia , Suínos/microbiologia , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia
15.
Transl Psychiatry ; 9(1): 91, 2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30770787

RESUMO

Phosphodiesterases (PDE) are key modulators of signal transduction and are involved in inflammatory cell activation, memory and cognition. There is a two-fold decrease in the expression of phosphodiesterase 8A (PDE8A) in the temporal cortex of major depressive disorder (MDD) patients. Here, we studied PDE8A mRNA-editing profile in two architectonically distinct neocortical regions in a clinically well-characterized cohort of age- and sex-matched non-psychiatric drug-free controls and depressed suicide decedents. By using capillary electrophoresis single-stranded conformational polymorphism (CE-SSCP), a previously validated technique to identify A-to-I RNA modifications, we report the full editing profile of PDE8A in the brain, including identification of two novel editing sites. Editing of PDE8A mRNA displayed clear regional difference when comparing dorsolateral prefrontal cortex (BA9) and anterior cingulate cortex (BA24). Furthermore, we report significant intra-regional differences between non-psychiatric control individuals and depressed suicide decedents, which could discriminate the two populations. Taken together, our results (i) highlight the importance of immune/inflammatory markers in major depressive disorder and suicide and (ii) establish a direct relationship between A-to-I RNA modifications of peripheral markers and A-to-I RNA editing-related modifications in brain. This work provides the first immune response-related brain marker for suicide and could pave the way for the identification of a blood-based biomarker that predicts suicidal behavior.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/genética , Transtorno Depressivo Maior/genética , Córtex Pré-Frontal/metabolismo , Edição de RNA/genética , RNA Mensageiro/metabolismo , Suicídio Consumado , Adolescente , Adulto , Autopsia , Estudos de Casos e Controles , Giro do Cíngulo/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo Conformacional de Fita Simples , Adulto Jovem
16.
Br Poult Sci ; 60(3): 187-194, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30686025

RESUMO

1. Two candidate genes, namely, Gonadotropin releasing hormone I (GnRHI) and Gonadotropin releasing hormone II (GnRHII) play pivotal roles in ovulation and egg production in chicken. The objective of this study was to explore polymorphism in these genes and to estimate the effects of polymorphism of these two genes on egg production and egg quality traits in White Leghorn laying hens. 2. Single strand conformation polymorphism followed by sequencing was performed to detect polymorphism in these genes. 3. The coding regions of the GnRHI and GnRHII genes were found to be polymorphic. In the GnRH1 gene, 12 haplotypes were determined, of which the h1 haplotype was predominant and the h5, h9 and h11 haplotypes were the least frequent ones. In the GnRHII gene, eight haplotypes were found, of which the h1 haplotype was the most frequent and the h6 was the least frequent haplotype in the White Leghorn population. 4. The haplogroups of GnRHI had a significant effect on body weight and egg production up to 64 weeks of age, yolk content, Haugh units and egg shell parameters. The h1h2 haplogroup of the GnRHI gene showed the highest egg production, with 211.0 ± 24.3 eggs up to 64 weeks of age, while the highest yolk content and Haugh unit was found in h3h10 haplogrouped birds. The haplogroups of GnRHII had a significant effect on age at sexual maturity (ASM) where the shortest ASM was found in the h1h4 birds (147.3 ± 5.9 d) and the longest ASM was observed in the h1h3 birds (160.6 ± 23.4 d). 5. It was concluded that GnRHI and GnRHII genes are polymorphic and have a significant effect on body weight, egg production and egg quality traits in White Leghorn laying hens.


Assuntos
Proteínas Aviárias/genética , Galinhas/fisiologia , Ovos/análise , Hormônio Liberador de Gonadotropina/análogos & derivados , Óvulo/fisiologia , Ácido Pirrolidonocarboxílico/análogos & derivados , Animais , Proteínas Aviárias/metabolismo , Galinhas/genética , Feminino , Hormônio Liberador de Gonadotropina/genética , Hormônio Liberador de Gonadotropina/metabolismo , Fases de Leitura Aberta , Polimorfismo Conformacional de Fita Simples , Ácido Pirrolidonocarboxílico/metabolismo , Análise de Sequência de DNA/veterinária
17.
Rev Gastroenterol Mex ; 84(2): 143-148, 2019.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-29898862

RESUMO

INTRODUCTION AND AIMS: Wilson's disease is characterized by the accumulation of copper in different organs, mainly affecting the liver, brain, and cornea, and is caused by mutations in the ATP7B gene. More than 120 polymorphisms in the ATP7B gene have been reported in the medical literature. The aim of the present study was to identify the conformational changes in the exon 3 region of the ATP7B gene and detect the p.L456V polymorphism in Cuban patients clinically diagnosed with Wilson's disease. MATERIAL AND METHODS: A descriptive study was conducted at the Centro Nacional de Genética Médica and the Instituto Nacional de Gastroenterología within the time frame of 2007-2012 and included 105 patients with a clinical diagnosis of Wilson's disease. DNA extraction was performed through the salting-out method and the fragment of interest was amplified using the polymerase chain reaction technique. The conformational shift changes in the exon 3 region and the presence of the p.L456V polymorphism were identified through the Single-Strand Conformation Polymorphism analysis. RESULTS: The so-called b and c conformational shift changes, corresponding to the p.L456V polymorphism in the heterozygous and homozygous states, respectively, were identified. The allelic frequency of the p.L456V polymorphism in the 105 Cuban patients that had a clinical diagnosis of Wilson's disease was 41% and liver-related symptoms were the most frequent in the patients with that polymorphism. CONCLUSION: The p.L456V polymorphism was identified in 64 Cuban patients clinically diagnosed with Wilson's disease, making future molecular study through indirect methods possible.


Assuntos
Degeneração Hepatolenticular/epidemiologia , Degeneração Hepatolenticular/genética , Adolescente , Adulto , Criança , ATPases Transportadoras de Cobre/genética , Cuba/epidemiologia , Éxons/genética , Feminino , Frequência do Gene , Humanos , Hepatopatias/epidemiologia , Hepatopatias/etiologia , Masculino , Pessoa de Meia-Idade , Mutação , Polimorfismo Genético/genética , Polimorfismo Conformacional de Fita Simples , Adulto Jovem
18.
Iran Biomed J ; 23(3): 220-7, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30053768

RESUMO

Background: KDM3A is a key epigenetic regulator expressed in the testis and is required for packaging and condensation of sperm chromatin. To this point, the association of the KDM3A gene with infertility has not been studied in human. The aim of this study was to screen any new mutation in KDM3A gene to explore more details of human male infertility. Methods: In this work, 150 infertile men (oligozoospermia and azoospermia) and 150 normal healthy fathers were studied. Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and sequencing were used to screen any mutation in exons 12, 22, and 24 of KDM3A. Results: The infertile men showed various SSCP patterns for the exons 12 and 24, but not for exon 22. A transversion point mutation in exon 12 and a single nucleotide deletion in exon 24 were detected using sequencing analysis. The transversion mutation was located in the preceding exon of lysine-specific demethylase1 and Jumonji (Jmj)-C domain and the later one (deletion) in the cupin-like motif of KDM3A protein. Neither Y chromosome microdeletions nor partial azoospermia factor deletion was found in these patients. Conclusion: The mutations found in infertile men with otherwise unexplained severe spermatogenic failure could be considered as the origin of their abnormalities.


Assuntos
Predisposição Genética para Doença , Infertilidade Masculina/genética , Histona Desmetilases com o Domínio Jumonji/genética , Mutação/genética , Adulto , Azoospermia/genética , Sequência de Bases , Deleção Cromossômica , Cromossomos Humanos Y/genética , Éxons/genética , Marcadores Genéticos , Humanos , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , Polimorfismo Conformacional de Fita Simples , Aberrações dos Cromossomos Sexuais , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/genética , Adulto Jovem
19.
J Anim Sci ; 97(2): 587-595, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30535023

RESUMO

The keratin-associated proteins (KAPs) are important constituents of wool fibers. Of the many mammalian KAP genes (KRTAPs) identified, KRTAP20-1 has been described in humans, but it has not been described in any other species. A search of the sheep genome using the human KRTAP20-1 sequence revealed a homologous open reading frame on chromosome 1, which would encode a high glycine-tyrosine KAP. PCR-single-stranded conformational polymorphism (PCR-SSCP) analysis identified 8 different banding patterns representing 8 unique DNA sequences (named A to H). The sequences had highest similarity to the human KRTAP20-1 sequence, and this suggests that they are variants of ovine KRTAP20-1. Among these variants, a 12-bp insertion/deletion and 6 single nucleotide poly- morphisms (SNPs), including one 5' untranslated region (UTR) SNP, one 3' UTR SNP, and 2 nonsynonymous SNPs, were detected. Variant A was found to be associated with a decrease in mean fiber diameter, fiber diameter standard deviation, and prickle factor, whereas variant C was associated with increased greasy fleece weight and decreased wool yield. These associations persisted after adjusting for the effect of 2 nearby KRTAPs (KRTAP6-3 and KRTAP22-1) that have also been reported to associate with these wool traits. This suggests that variation in KRTAP20-1 affects wool yield and mean fiber diameter-associated traits, and that this effect is unlikely to be the result of the clustering of these KRTAPs on chromosome 1.


Assuntos
Genoma/genética , Queratinas/genética , Ovinos/genética , Animais , Sequência de Bases , Glicina/genética , Fenótipo , Filogenia , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Nucleotídeo Único , Polimorfismo Conformacional de Fita Simples , Alinhamento de Sequência/veterinária , Ovinos/fisiologia , Tirosina/genética ,
20.
Colomb Med (Cali) ; 50(3): 163-175, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-32284662

RESUMO

Introduction: Breast cancer is the most common neoplasia of women from all over the world especially women from Colombia. 5%-10% of all cases are caused by hereditary factors, 25% of those cases have mutations in the BRCA1/BRCA2 genes. Objective: The purpose of this study was to identify the mutations associated with the risk of familial breast and/or ovarian cancer in a population of Colombian pacific. Methods: 58 high-risk breast and/or ovarian cancer families and 20 controls were screened for germline mutations in BRCA1 and BRCA2, by Single Strand Conformation Polymorphism (SSCP) and sequencing. Results: Four families (6.9%) were found to carry BRCA1 mutations and eight families (13.8%) had mutations in BRCA2. In BRCA1, we found three Variants of Uncertain Significance (VUS), of which we concluded, using in silico tools, that c.81-12C>G and c.3119G>A (p.Ser1040Asn) are probably deleterious, and c.3083G>A (p.Arg1028His) is probably neutral. In BRCA2, we found three variants of uncertain significance: two were previously described and one novel mutation. Using in silico analysis, we concluded that c.865A>G (p.Asn289Asp) and c.6427T>C (p.Ser2143Pro) are probably deleterious and c.125A>G (p.Tyr42Cys) is probably neutral. Only one of them has previously been reported in Colombia. We also identified 13 polymorphisms (4 in BRCA1 and 9 in BRCA2), two of them are associated with a moderate increase in breast cancer risk (BRCA2 c.1114A>C and c.8755-66T>C). Conclusion: According to our results, the Colombian pacific population presents diverse mutational spectrum for BRCA genes that differs from the findings in other regions in the country.


Assuntos
Proteína BRCA1/genética , Proteína BRCA2/genética , Neoplasias da Mama/genética , Neoplasias Ovarianas/genética , Adulto , Idoso , Estudos de Casos e Controles , Colômbia , Simulação por Computador , Feminino , Predisposição Genética para Doença , Mutação em Linhagem Germinativa , Humanos , Pessoa de Meia-Idade , Polimorfismo Conformacional de Fita Simples
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