scholarly journals Epigenetic characterization of the H19/IGF2 locus in calf clones placenta

2020 ◽  
Vol 40 (12) ◽  
pp. 1063-1072
Author(s):  
Natalia S. Costa ◽  
Márcia M. Silveira ◽  
Luna N. Vargas ◽  
Alexandre R. Caetano ◽  
Rodolfo Rumpf ◽  
...  
Keyword(s):  
Methylation Pattern ◽  
Significant Difference ◽  
Pcr Products ◽  
H19 Gene ◽  
Imprinting Control Region ◽  
Low Efficiency ◽  
Genomic Regions ◽  
Methylation Patterns ◽  
Exon 10

ABSTRACT: Somatic Cell Nuclear Transfer (SCNT-Cloning) is a promising technique in many areas and is based on genetically identical individuals. However, its efficiency is low. Studies suggest that the leading cause is inadequate epigenetic reprogramming. This study aimed to characterize the methylation pattern of the exon 10 regions of the IGF2 gene and the Imprinting Control Region (ICR) of the H19 gene in the placenta of cloned calves. For this study, female and male cloned calves presenting different phenotypes were used. Genomic DNA from these animals’ placenta was isolated, then treated with sodium bisulfite and amplified to the ICR/H19 and IGF2 loci. PCR products were cloned into competent bacteria and finally sequenced. A significant difference was found between controls and clones with healthy phenotypes for the ICR/H19 region. In this region, controls showed a hemimethylated pattern, as predicted in the literature due to this region has an imprinted control, while clones were showed less methylated. For the IGF2, no significant differences were found between controls and clones. These results suggest that different genomic regions in the genome may be independently reprogrammed and that failures in reprogramming the DNA methylation patterns of imprinted genes may be one of the causes of the low efficiency of SCNT.

Characterization of expression and DNA methylation patterns of H19 gene in human sertoli cells (HSEC LINE)

2012 ◽  
Vol 98 (3) ◽  
pp. S143
Author(s):  
M.A. Ribeiro ◽  
M.B. dos Reis ◽  
F.D. Oliveira ◽  
C. Briton-Jones ◽  
C.A. Rainho ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Sertoli Cells ◽  
H19 Gene ◽  
Methylation Patterns

scholarly journals A 4-Year-Old Boy with Beckwith Wiedemann Syndrome (BWS)

PRILOZI ◽  
2018 ◽  
Vol 39 (2-3) ◽  
pp. 131-135
Author(s):  
Aleksandra Janchevska ◽  
Velibor Tasic ◽  
Nevenka Laban ◽  
Momir Polenakovic ◽  
Zoran Gucev ◽  
...  
Keyword(s):  
Clinical Presentation ◽  
Molecular Genetic ◽  
Methylation Pattern ◽  
Molecular Genetic Analysis ◽  
Differentially Methylated Region ◽  
Embryonal Tumors ◽  
Left And Right ◽  
Imprinting Control Region ◽  
Chromosome 11P15.5

Abstract Objectives: Molecular characterization of a patient with BWS. Clinical presentation and intervention: A 4-year-old boy with overgrowth (weight above 99th and height at 99th percentile) had longitudinal hemihypertrophy of the tongue and left cheek. In addition, there was a difference of one centimeter in the circumference of the left and right leg. Molecular genetic analysis revealed hypomethylation of KvDRM1 (LIT1) in the imprinting control region-2 (ICR2) on chromosome 11p15.5 and a normal methylation pattern of the H19-differentially methylated region (H19-DMR) in the ICR1. The estimated tumor risk was 1-5%. Conclusion: This patient with clinical characteristics of BWS has an imprinting defect associated with a low risk of embryonal tumors.

scholarly journals A model for the aberrant DNA methylomes in aging cells and cancer cells

2019 ◽  
Vol 47 (4) ◽  
pp. 997-1003 ◽  
Author(s):  
Huiming Zhang ◽  
Kang Zhang ◽  
Jian-Kang Zhu
Keyword(s):  
Dna Methylation ◽  
Cancer Cells ◽  
Developmental Stages ◽  
Methylation Pattern ◽  
Epigenetic Mark ◽  
Dna Hypomethylation ◽  
Genome Wide ◽  
Abnormal Cells ◽  
Genomic Regions ◽  
Methylation Patterns

Abstract DNA methylation at the fifth position of cytosine is a major epigenetic mark conserved in plants and mammals. Genome-wide DNA methylation patterns are dynamically controlled by integrated activities of establishment, maintenance, and removal. In both plants and mammals, a pattern of global DNA hypomethylation coupled with increased methylation levels at some specific genomic regions arises at specific developmental stages and in certain abnormal cells, such as mammalian aging cells and cancer cells as well as some plant epigenetic mutants. Here we provide an overview of this distinct DNA methylation pattern in mammals and plants, and propose that a methylstat, which is a cis-element responsive to both DNA methylation and active demethylation activities and controlling the transcriptional activity of a key DNA methylation regulator, can help to explain the enigmatic DNA methylation patterns in aging cells and cancer cells.

scholarly journals Morphological, Molecular and Genomic Characterization of Two Inter-Subspecific Hybrids between Olive Cultivars and Olive Subspecies

Horticulturae ◽  
2021 ◽  
Vol 7 (6) ◽  
pp. 138
Author(s):  
Jinhua Li ◽  
Xinyue Ji ◽  
Zhaoshan Wang ◽  
Yanfei Zeng ◽  
Jianguo Zhang
Keyword(s):  
Southern China ◽  
Single Copy ◽  
F1 Hybrids ◽  
Ssr Analysis ◽  
Significant Difference ◽  
Olive Cultivars ◽  
Pcr Products ◽  
Genomic Characterization ◽  
Alignment Analysis

Two inter-subspecific F1 hybrids have been obtained by crossing olive cultivars (‘Frantoio’ and ‘Coratina’) with pollen donors from olive subspecies (Olea europaea subsp. cuspidata) to enrich the germplasm of cultivated olive in southern China. This study aimed to investigate the characterization of morphological traits and molecular markers in the two hybrids and their parents of crosses. The morphological study showed a significant difference between genotypes according to the main discriminative parameters on qualitative and quantitative traits of leaf, fruit, and endocarp. A set of six co-dominant polymorphic simple sequence repeats (SSRs) were used for molecular identification, and SSR analysis confirmed that two progenies were the offspring of their cited parents based on the presence of parental specific SSR alleles. Three single-copy nuclear loci (SCNL) primer pairs were used for amplification of single-copy genes in the two progenies and their parents and after then PCR products were sequenced. Sequence alignment analysis on the effective data showed a total of 15 different base sites between two progenies, which were confirmed as true inter-specific hybrids between olive cultivars and subsp. cuspidata.

Examination of the dynamics of global DNA methylation pattern establishment during spermatogenesiss

2007 ◽  
Vol 30 (4) ◽  
pp. 90
Author(s):  
Kirsten Niles ◽  
Sophie La Salle ◽  
Christopher Oakes ◽  
Jacquetta Trasler
Keyword(s):  
Dna Methylation ◽  
Germ Cell ◽  
Germ Cells ◽  
Epigenetic Modification ◽  
Methylation Pattern ◽  
Chromosome 9 ◽  
Specific Gene ◽  
Global Methylation ◽  
Dna Methylation Pattern ◽  
Methylation Patterns

Background: DNA methylation is an epigenetic modification involved in gene expression, genome stability, and genomic imprinting. In the male, methylation patterns are initially erased in primordial germ cells (PGCs) as they enter the gonadal ridge; methylation patterns are then acquired on CpG dinucleotides during gametogenesis. Correct pattern establishment is essential for normal spermatogenesis. To date, the characterization and timing of methylation pattern acquisition in PGCs has been described using a limited number of specific gene loci. This study aimed to describe DNA methylation pattern establishment dynamics during male gametogenesis through global methylation profiling techniques in a mouse model. Methods: Using a chromosome based approach, primers were designed for 24 regions spanning chromosome 9; intergenic, non-repeat, non-CpG island sequences were chosen for study based on previous evidence that these types of sequences are targets for testis-specific methylation events. The percent methylation was determined in each region by quantitative analysis of DNA methylation using real-time PCR (qAMP). The germ cell-specific pattern was determined by comparing methylation between spermatozoa and liver. To examine methylation in developing germ cells, spermatogonia from 2 day- and 6 day-old Oct4-GFP (green fluorescent protein) mice were isolated using fluorescence activated cell sorting. Results: As compared to liver, four loci were hypomethylated and five loci were hypermethylated in spermatozoa, supporting previous results indicating a unique methylation pattern in male germ cells. Only one region was hypomethylated and no regions were hypermethylated in day 6 spermatogonia as compared to mature spermatozoa, signifying that the bulk of DNA methylation is established prior to type A spermatogonia. The methylation in day 2 spermatogonia, germ cells that are just commencing mitosis, revealed differences of 15-20% compared to day 6 spermatogonia at five regions indicating that the most crucial phase of DNA methylation acquisition occurs prenatally. Conclusion: Together, these studies provide further evidence that germ cell methylation patterns differ from those in somatic tissues and suggest that much of methylation at intergenic sites is acquired during prenatal germ cell development. (Supported by CIHR)

Linkage between H. pylori Infection and TNF-α polymorphism in The Pregnant Women

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Ghaidaa Raheem Lateef ◽  
Azhar Omaran Al-Thahab
Keyword(s):  
Pregnant Women ◽  
Serum Antibody ◽  
Rapid Test ◽  
Negative Group ◽  
Positive Group ◽  
Gynecology And Obstetrics ◽  
Tnf Α ◽  
Significant Difference ◽  
Pcr Products ◽  
H Pylori

A study was performed on 100 pregnant women in the outpatient department of gynecology and obstetrics of Maternity and Children Hospital in Al-Diwaniya City during the period between (March to September 2016). One hundred blood samples (50 for patients and 50 for control) were collected under the supervision of the treating gynecologist. The detection of Helicobacter. pylori was done by the use of the serum antibody Rapid test. The results showed that 50 (100%) were positive and 50 (100%) were negative for H. pylori in above method.All blood of patients and control samples were used for the extraction of genomic DNA,where the 107 bp PCR product size. Genotyping of the TNF-α-308 SNP (G/A)was performed by restriction fragment length polymorphism PCR (RFLP-PCR). PCR products were digested with restr NcoI iction enzyme. Individuals with the TNF-α-308(GG) homozygote produced digested DNA bands at 80,and 20 bp bp. A heterozygous genotype ofTNF-α-308 (GA)produced 107 bp,80 bp,and 20 bp bands. Individuals with the TNF-α-308 (AA) homozygote genotype had no amplicon digested and generated only one band of 107 bp. There was a significant difference in the frequency of the TNF-α-308(GG)genotype between H. pylori positive group and H. pylori negative group(72%,78% respectively). Also for GA genotype,there was a significant difference between H. pylori positive group and H. pylori negative group(24%,18% respectively). Concerning the frequency of the TNF-α-308 (AA)genotype between H. pylori positive group and H. pylori negative group,there was no significant difference between the two groups.

MICROSCOPIC AND MOLECULAR DIAGNOSIS OF Ascaridia spp. IN DOMESTIC PIGEONS(Columba livia domestica) IN BAGHDAD CITY,IRAQ

2020 ◽  
Vol 51 (4) ◽  
pp. 1220-1225
Author(s):  
Faraj & Al- Amery
Keyword(s):  
Molecular Diagnosis ◽  
Columba Livia ◽  
Ascaridia Galli ◽  
Molecular Assay ◽  
Infection Rates ◽  
Conventional Pcr ◽  
Significant Difference ◽  
Pcr Products ◽  
Males And Females ◽  
Domestic Pigeons

Ascaridiosis is a very important parasitic disease of birds, it is caused by Ascaridia. This study was conducted to identify the Ascaridia species by microscopic and molecular assay in Baghdad city. One hundred and sixty fecal samples were collected from domestic pigeons during the period from 1/1/ 2019 to 31/3/ 2019.  Results showed that the rate of infection for Ascaridia spp. 15.62% by microscopic examination.  Significant difference was observed in infection rates between males and females pigeons. Fifty samples randomly selected and subjected to molecular diagnosis of Ascaridia  spp.. Molecular examination results, the total infection rate showed 16%(8/50). The eight  positive PCR products were sequenced and deposited in Gene bank data base, phylogenic analysis demonstrated that 4 sequences belongs to Ascaridia galli ( MK918635.1, MK918636.1, MK918847.1, MK919081.1), while 2 (MK919199.1, MK919200.1) belong to  Ascaridia nymphii and 2 (MK919207.1, MK919264.1)  belong to Ascaridia numidae. It is the first study in Iraq to diagnosis of  Ascaridia nymphii and Ascaridia numidae  in domesticed pigeons by using conventional PCR.

Caracterização da Peneira Média em Clones de Coffea canephora

REVISTA FIMCA ◽  
2018 ◽  
Vol 5 (2) ◽  
pp. 28-31
Author(s):  
Darlan Darlan Sanches Barbosa Alves ◽  
Victor Mouzinho Spinelli ◽  
Marcos Santana Moraes ◽  
Carolina Augusto De Souza ◽  
Rodrigo da Silva Ribeiro ◽  
...  
Keyword(s):  
Genetic Improvement ◽  
Block Design ◽  
Coffea Canephora ◽  
Coffee Production ◽  
Randomized Block Design ◽  
Significant Difference ◽  
The Mean ◽  
Four Blocks ◽  
Selection Of

Introdução: O estado de Rondônia se destaca como tradicional produtor de café, sendo o segundo maior produtor brasileiro de C. canephora. No melhoramento genético de C. canephora, a seleção de plantas de elevada peneira média está associada à bebida de qualidade superior. Objetivos: O objetivo desse estudo foi avaliar a variabilidade genética de clones de C. canephora para o tamanho dos grãos, mensurado a partir da avaliação da peneira média (PM). Materiais e Métodos: Para isso, foi conduzido ao longo de dois anos agrícolas experimento no campo experimental da Embrapa no município de Ouro Preto do Oeste-RO, para a avaliação da peneira média de 130 genótipos (clones) com características das variedades botânicas Conilon, Robusta e híbridos intervarietais. O delineamento experimental utilizado foi de blocos ao acaso, com quatro repetições de quatro plantas por parcela. Resultados: Não houve resultados significativos para a interação clones X anos, indicando uma maior consistência no comportamento das plantas ao longo do tempo. Porém foram observadas diferenças significativas para o tamanho dos grãos entre os genótipos avaliados, possibilitando selecionar genótipos superiores. Conclusão: Os genótipos agruparam-se em cinco classes de acordo com o teste de média, subsidiando a caracterização de um gradiente de variabilidade da característica avaliada ABSTRACTIntroduction: Coffea canephora accounts for approximately 35% of the world's coffee production. The state of Rondônia stands out as a traditional coffee producer, being the second largest Brazilian producer of C. canephora. In the classical genetic improvement of C. anephora, the selection of plants of high average sieve is associated with a drink of superior quality. Objectives: The objective of this udy was to evaluate the genetic variability of Coffea canephora clones for the agronomic medium sieve (PM). Materials and Methods: The experiment was conducted in the experimental field of Embrapa, municipality of OuroPreto do Oeste-RO, located at coordinates 10º44'53 "S and 62º12'57". One hundred thirty genotypes (clones) of botanical characteristics Conilon, Robusta and intervarietal hybrids were evaluated in the agricultural years 2013-2014 and 2014-2015. The experimental design was a randomized block design with four blocks and four plants per plot, spacing 3.5 x 1.5 meters between plants. Results: Significant difference was found for the grain size. According to the F test, at 5% probability, the genotypes were grouped into five classes according to the mean test. Conclusion: The results obtained subsidized the characterization of a variability gradient of the evaluated trait.

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