High levels of mitochondrial heteroplasmy modify the development of ovine - bovine interspecies nuclear transferred embryos

2012 ◽  
Vol 24 (3) ◽  
pp. 501 ◽  
Author(s):  
Song Hua ◽  
Chenglong Lu ◽  
Yakun Song ◽  
Ruizhe Li ◽  
Xu Liu ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Control Group ◽  
Bisulfite Genomic Sequencing ◽  
Mitochondrial Suspension ◽  
Mitochondrial Heteroplasmy ◽  
Polymerase Chain ◽  
Early Developmental ◽  
Bovine Oocytes

To investigate the effect of mitochondrial heteroplasmy on embryo development, cloned embryos produced using bovine oocytes as the recipient cytoplasm and ovine granulosa cells as the donor nuclei were complemented with 2 pL mitochondrial suspension isolated from ovine (BOOMT embryos) or bovine (BOBMT embryos) granulosa cells; cloned embryos without mitochondrial injection served as the control group (BO embryos). Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) and sodium bisulfite genomic sequencing were used to analyse mRNA and methylation levels of pluripotency genes (OCT4, SOX2) and mitochondrial genes (TFAM, POLRMT) in the early developmental stages of cloned embryos. The number of mitochondrial DNA copies in 2 pL ovine-derived and bovine-derived mitochondrial suspensions was 960 ± 110 and 1000 ± 120, respectively. The blastocyst formation rates were similar in BOBMT and BO embryos (P > 0.05), but significantly higher than in BOOMT embryos (P < 0.01). Expression of OCT4 and SOX2, as detected by RT-qPCR, decreased significantly in BOOMT embryos (P < 0.05), whereas the expression of TFAM and POLRMT increased significantly, compared with expression in BOOMT and BO embryos (P < 0.05). In addition, methylation levels of OCT4 and SOX2 were significantly greater (P < 0.05), whereas those of TFAM and POLRMT were significantly lower (P < 0.01), in BOOMT embryos compared with BOBMT and BO embryos. Together, the results of the present study suggest that the degree of mitochondrial heteroplasmy may affect embryonic development.

Quantitative PCR Analysis for Bcl-2/IgH in a Phase III Study of Yttrium-90 Ibritumomab Tiuxetan As Consolidation of First Remission in Patients With Follicular Lymphoma

2009 ◽  
Vol 27 (36) ◽  
pp. 6094-6100 ◽  
Author(s):  
Lindsey Goff ◽  
Karin Summers ◽  
Sameena Iqbal ◽  
Jens Kuhlmann ◽  
Michael Kunz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Follicular Lymphoma ◽  
Quantitative Polymerase Chain Reaction ◽  
Phase Iii ◽  
Pcr Analysis ◽  
Control Group ◽  
Ibritumomab Tiuxetan ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Yttrium 90

Purpose The randomized First-Line Indolent Trial (FIT) was conducted in patients with advanced follicular lymphoma (FL), to evaluate the safety and efficacy of yttrium-90 (90Y) ibritumomab tiuxetan given as consolidation of complete or partial remission. This study of minimal residual disease was undertaken in parallel, to determine the rate of conversion from bcl-2 polymerase chain reaction (PCR) –detectable to –undetectable status and the corresponding effect on progression-free survival (PFS). Patients and Methods Blood samples from 414 patients (90Y-ibritumomab, n = 208; control, n = 206) were evaluated using real-time quantitative polymerase chain reaction (RQ-PCR); 186 were found to have the bcl-2 rearrangement and were thus eligible for inclusion in the RQ-PCR analysis. Results Overall, 90% of treated patients converted from bcl-2 PCR–detectable to –undetectable disease status, compared with 36% in the control group. Treatment significantly prolonged median PFS in patients converting to bcl-2 PCR-undetectable status (40.8 v 24.0 months in the control group; P < .01, hazard ratio [HR], 0.399). In patients who had bcl-2 PCR-detectable disease at random assignment, treatment significantly prolonged median PFS (38.4 v 8.2 months in the control group; P < .01, HR, 0.293). Conclusion Eradication of PCR-detectable disease occurred more frequently after treatment with 90Y-ibritumomab tiuxetan and was associated with prolongation of PFS.

scholarly journals Supraphysiological Concentrations of Bisphenol A Alter the Expression of Extracellular Vesicle-Enriched miRNAs From Human Primary Granulosa Cells

2019 ◽  
Vol 169 (1) ◽  
pp. 5-13 ◽  
Author(s):  
Rodosthenis S Rodosthenous ◽  
Andrea A Baccarelli ◽  
Abdallah Mansour ◽  
Michal Adir ◽  
Ariel Israel ◽  
...  
Keyword(s):  
Bisphenol A ◽  
Granulosa Cells ◽  
Target Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Extracellular Vesicle ◽  
Biological Mechanisms ◽  
Expression Levels ◽  
Biologically Relevant ◽  
Reproductive Effects ◽  
Polymerase Chain

Abstract Bisphenol A (BPA) is a widely used chemical that has been detected in follicular fluid and associated with adverse reproductive effects. Granulosa cells have an important role in follicular growth and oocyte maturation, however, little is known about the biological mechanisms of BPA toxicity on human granulosa cells. In this study, we exposed primary granulosa cells to different concentrations of BPA (0, 20, 200, 2000, and 20 000 ng/ml) and used quantitative polymerase chain reaction to measure the expression levels of miRNAs enriched in extracellular vesicles (EV-enriched miRNAs), and cellular levels of selected target genes of differentially expressed EV-enriched miRNAs. We found that exposure to 20 000 ng/ml BPA was associated with decreased levels of EV-miR-27b-3p (FC = 0.58, p = .04) and increased levels of its biologically relevant target genes FADD (FC = 1.22, p = .01), IGF1 (FC = 1.59, p = .06), and PPARG (FC = 1.73, p = .001) as compared with the control. In addition, we observed that under the same exposure conditions, the expression levels of miR-27b-3p in granulosa cells were also downregulated (FC = 0.65, p = .03) as compared with the control. Our findings suggest that both cellular and extracellular changes in gene expression may mediate BPA toxicity in granulosa cells.

scholarly journals Expression and distribution of EPHA4 and Ephrin A3 in Aohan fine-wool sheep skin

2022 ◽  
Vol 65 (1) ◽  
pp. 11-19
Author(s):  
Yu Cui ◽  
Chunliang Wang ◽  
Lirong Liu ◽  
Nan Liu ◽  
Jianning He
Keyword(s):  
Hair Follicle ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Dermal Papilla ◽  
Hair Follicles ◽  
Fine Hair ◽  
Dermal Papilla Cells ◽  
Temporal Specificity ◽  
Polymerase Chain ◽  
Hair Follicle Development

Abstract. The objective of this study was to identify the expression and distribution of EPHA4 and Ephrin A3 genes in the development and morphogenesis of hair follicles in fine-wool sheep. The results could lay a theoretical basis for understanding the molecular mechanism that regulates hair follicle development. The skin of Aohan fine-wool sheep at different developmental stages (embryonic day 90, E90d, and 120, E120d, and postnatal day 1, B1d, and 30, B30d) were selected. Real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry were used to study the levels of mRNA and proteins, respectively. The RT-qPCR results showed that the mRNA expression level of EPHA4 at B1d was significantly lower than at E120d (p<0.01). The expression of Ephrin A3 at E120d was significantly higher than that at E90d and B1d (p<0.01). Immunohistochemical detection results showed that the level and localisation of EPHA4 and Ephrin A3 proteins had spatial and temporal specificity. EPHA4 expression in dermal papilla cells might be important for inducing Aohan fine-hair follicle regeneration and for controlling the properties of the hair. Ephrin A3 might play an important role in the redifferentiation of secondary hair follicles and might also be involved in the inhibition of apoptosis-related gene expression in hair follicles. The Ephrin A3 signalling pathway might accelerate the growth of fine-hair follicles and increase the density of hair follicles.

Effect of 2-(4-chlorophenylthio) ethylamine hydrochloride on lycopene biosynthesis in pepper fruits

2020 ◽  
pp. 1-6
Author(s):  
Li Li ◽  
Shi-Lin Tian ◽  
Shi-Lun Xu
Keyword(s):  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Inhibitory Effect ◽  
Development Stage ◽  
Metabolic Product ◽  
Normal Expression ◽  
Other Substances ◽  
Lycopene Content ◽  
Polymerase Chain

Lycopene is an intermediate metabolic product of the capsanthin biosynthesis pathway in pepper fruits and is one of the strongest antioxidants found in plants. During the ripening of pepper fruits, lycopene is almost completely transformed into the downstream metabolic product capsanthin as well as other substances. As a result, lycopene cannot be enriched in ripe pepper fruits; however, the lycopene content can be increased by 2-(4-chlorophenylthio) ethylamine hydrochloride (CPTA) treatment, using the optimal concentration at the optimal development stage of pepper fruits. The current study tested different CPTA concentrations and fruit developmental stages to increase the lycopene content in pepper fruits. The results showed that the lycopene content was significantly enriched in pepper fruits treated with 0.1% CPTA applied at the turning stage. Real-time quantitative polymerase chain reaction analysis showed that CPTA treatment significantly promoted the expression of the upstream genes (Psy and PDS) involved in the anabolic metabolism of lycopene; however, CPTA treatment had a significant inhibitory effect on the downstream gene (Lcyb) of lycopene synthesis. Therefore, in pepper fruits, CPTA inhibits the normal expression of the Lcyb gene downstream of lycopene, thus achieving notable lycopene enrichment.

scholarly journals Down-regulation of miR-10a-5p promotes proliferation and restricts apoptosis via targeting T-box transcription factor 5 in inflamed synoviocytes

2018 ◽  
Vol 38 (2) ◽  
Author(s):  
Nazim Hussain ◽  
Wenhua Zhu ◽  
Congshan Jiang ◽  
Jing Xu ◽  
Manman Geng ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Transcription Factor ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Counting ◽  
Control Group ◽  
Down Regulation ◽  
T Box ◽  
Proliferation And Apoptosis ◽  
Polymerase Chain

Synoviocytes from rheumatoid arthritis (RA) patients share certain features with tumor cells, such as over proliferation and invasion. Anomalous microRNA (miRNA) expression may participate in the pathogenesis of RA in different ways. The objective of the present study was to observe the role of miR-10a-5p targeting T-box transcription factor 5 (TBX5) gene on synoviocyte proliferation and apoptosis in RA. Human synovial sarcoma cell line, SW982 cells stimulating with interleukin-1β (IL-1β) were transfected with miR-10a-5p mimic and siRNA of TBX5. The real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting analysis were used to evaluate the expression level of miR-10a-5p and TBX5 in SW982 cells respectively. Further, the proliferation and apoptosis of SW982 cells after treatment were determined by cell counting kit (CCK-8) and flow cytometry analysis respectively. We found that the miR-10a-5p showed down-regulated while TBX5 showed up-regulated expression in synoviocytes after stimulation with IL-1β. The miR-10a-5p mimic treatment showed a decline in cell proliferation while the increased rate of cell apoptosis as compared with control. Moreover, knockdown of TBX5 favored the apoptosis and reduced the cell proliferation as compared with control group. We conclude that down-regulation of miR-10a-5p promotes proliferation and restricts apoptosis via targeting TBX5 in inflamed synoviocytes.

Decreased expression of fibulin-4 in aortic wall of aortic dissection

Vascular ◽  
2013 ◽  
Vol 22 (1) ◽  
pp. 35-41 ◽  
Author(s):  
P Huawei ◽  
C Qian ◽  
T Chuan ◽  
L Lei ◽  
W Liang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aortic Dissection ◽  
Aortic Wall ◽  
Quantitative Polymerase Chain Reaction ◽  
Artery Bypass ◽  
Elastic Fiber ◽  
Elastic Fibers ◽  
Control Group ◽  
Chain Reaction ◽  
Polymerase Chain

In this research, we will examine the expression of Fibulin-4 in aortic wall to find out its role in aortic dissection development. The samples of aortic wall were obtained from 10 patients operated for acute ascending aortic dissection and five patients for chronic ascending aortic dissection. Another 15 pieces of samples from patients who had coronary artery bypass were as controls. The aortic samples were stained with aldehyde magenta dyeing to evaluate the arrangement of elastic fibers. The Fibulin-4 protein and mRNA expression were both determined by Western blot and realtime quantitative polymerase chain reaction. Compared with the control group, both in acute and chronic ascending aortic dissection, elastic fiber fragments increased and the expression of fibulin-4 protein significantly decreased ( P = 0.045 < 0.05). The level of fibulin-4 mRNA decreased in acute ascending aortic dissection ( P = 0.034 < 0.05), while it increased in chronic ascending aortic dissection ( P = 0.004 < 0.05). The increased amounts of elastic fiber fragments were negatively correlated with the expression of fibulin-4 mRNA in acute ascending aortic dissection. In conclusion, in aortic wall of ascending aortic dissection, the expression of fibulin-4 protein decreased and the expression of fibulin-4 mRNA was abnormal. Fibulin-4 may play an important role in the pathogenesis of aortic dissection.

scholarly journals Genetic Polymorphisms of Metabolic Enzyme Genes Associated with Leukocyte Mitochondrial DNA Copy Number in PAHs Exposure Workers

2020 ◽  
Author(s):  
Xinling Li ◽  
Xiaoran Duan ◽  
Hui Zhang ◽  
Mingcui Ding ◽  
Yanbin Wang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Polymorphisms ◽  
Peripheral Blood ◽  
Quantitative Polymerase Chain Reaction ◽  
Metabolic Enzymes ◽  
Control Group ◽  
Peripheral Blood Leukocytes ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Polymerase Chain

Abstract Background: PAHs exposure had been reported to be a risk factor of mtDNAcn in our early study. However, the effect of metabolic enzymes’ genetic polymorphisms on mtDNAcn in PAHs-Exposure workers has not been fully evaluated.Methods: We investigated the effects of metabolic enzymes’ genetic polymorphisms on mtDNAcn among 544 coke oven workers and 238 office staffs. The mtDNAcn of peripheral blood leukocytes was measured using Real-time quantitative polymerase chain reaction method. Polymerase chain reaction and restriction fragment length was used to detect five polymorphisms in GSTT1, GSTM1, GSTP1 rs1695, CYP2E1 rs6413432, and CYP2E1 rs3813867.Results: The mtDNAcn in peripheral blood leukocytes was significantly lower in the exposure group than that in the control group (P < 0.001). The 1-OHPYR had an increasing trend with the genotypes AA→AG→GG of GSTP1 rs1695 in the control group. Generalized linear model indicated that the influencing factors of mtDNAcn were PAHs-exposure [b(95% CI) = -0.420 (-0.469, -0.372), P < 0.001], male [β(95% CI) = -0.058 (-0.103, -0.012), P = 0.013] ,and AA genotype for GSTP1 rs1695 [β(95% CI) = -0.051 (-0.095, -0.008), P = 0.020].Conclusions: The male was susceptibility to PAHs exposure. The AA genotype of GSTP1 rs1695 may influence the toxicity of PAHs and associated with the decreased of mtDNAcn.

scholarly journals Relationship between Expression of Plasma lncRNA-HEIH and Prognosis in Patients with Coronary Artery Disease

2021 ◽  
Vol 2021 ◽  
pp. 1-5
Author(s):  
Zhenying Zhang ◽  
Sushuang Nan ◽  
Xiujuan Duan ◽  
Lizhong Wang ◽  
Xiaojing Sun ◽  
...  
Keyword(s):  
Coronary Artery Disease ◽  
Coronary Artery ◽  
Adverse Events ◽  
Noncoding Rna ◽  
Quantitative Polymerase Chain Reaction ◽  
Control Group ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Artery Disease ◽  
Cardiac Adverse Events

Objective. We aimed to investigate the expression of long noncoding RNA- (lncRNA-) HEIH in patients with coronary artery disease (CAD) and its impact on patients’ prognosis. Patients and Methods. From July 2015 to December 2018, 250 patients who underwent coronary angiography, including 50 in the control group and 150 in the CAD group, were collected for detection of the expression of lncRNA-HEIH by real-time quantitative polymerase chain reaction (qPCR). The severity of CAD was evaluated through SYNTAX scoring system. In addition, these patients with CAD were followed up for 3 years, and the major cardiac adverse events such as myocardial infarction and revascularization were recorded. Results. The expression of lncRNA-HEIH in plasma of patients with CAD was remarkably higher than that in the control subjects and was verified to be relevant to the severity of CAD. Meanwhile, it was found that CAD patients with high expression of lncRNA-HEIH had higher rates of dyslipidemia as well as CAD family history and higher overall incidence of major cardiac adverse events than those with low expression of lncRNA-HEIH. Conclusions. lncRNA-HEIH expression is upregulated in the plasma of CAD patients, which is capable of affecting the prognosis of patients.

scholarly journals Identification and Characterization of Osmoregulation Related MicroRNAs in Gills of Hybrid Tilapia Under Three Types of Osmotic Stress

2021 ◽  
Vol 12 ◽  
Author(s):  
Huanhuan Su ◽  
Jiajia Fan ◽  
Dongmei Ma ◽  
Huaping Zhu
Keyword(s):  
Polymerase Chain Reaction ◽  
Osmotic Stress ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Protein Translation ◽  
Control Group ◽  
Alkali Stress ◽  
Chain Reaction ◽  
Polymerase Chain

Researchers have increasingly suggested that microRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression and protein translation in organs and respond to abiotic and biotic stressors. To understand the function of miRNAs in osmotic stress regulation of the gills of hybrid tilapia (Oreochromis mossambicus ♀ × Oreochromis urolepis hornorum ♂), high-throughput Illumina deep sequencing technology was used to investigate the expression profiles of miRNAs under salinity stress (S, 25‰), alkalinity stress (A, 4‰) and salinity–alkalinity stress (SA, S: 15‰, A: 4‰) challenges. The results showed that 31, 41, and 27 upregulated and 33, 42, and 40 downregulated miRNAs (P &lt; 0.05) were identified in the salt stress, alkali stress, and saline–alkali stress group, respectively, which were compared with those in the control group (C). Fourteen significantly differently expressed miRNAs were selected randomly and then validated by a quantitative polymerase chain reaction. On the basis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, genes related to osmoregulation and biosynthesis were enriched in the three types of osmotic stress. In addition, three miRNAs and three predicted target genes were chosen to conduct a quantitative polymerase chain reaction in the hybrid tilapia and its parents during 96-h osmotic stress. Differential expression patterns of miRNAs provided the basis for research data to further investigate the miRNA-modulating networks in osmoregulation of teleost.

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