84 Aging Model in Pig Oocytes as Viewed Through DNA Epigenetic Modification

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 108-109
Author(s):  
Haley A Arena ◽  
Kimberly Sprungl ◽  
Skyla Reynolds ◽  
Brian D Whitaker
Keyword(s):  
In Vitro Maturation ◽  
In Vitro Model ◽  
Time Window ◽  
Epigenetic Modification ◽  
Trichostatin A ◽  
Epigenetic Modifications ◽  
Optimal Time ◽  
Vitro Model ◽  
Effects Of Aging

Abstract Oocytes of older animals are less likely to be fertilized during the optimal time window post ovulation, resulting in the potential diminished fertilization and embryonic development success. The activity of the epigenetic modifications during this period is a possible target to reverse these damaging effects of aging. The objective of this study was to study the effects of aging during in vitro oocyte maturation in pigs on epigenetic modifications. Oocytes (n = 54) were matured with or without Trichostatin A (TSA; 100 ng/mL), a known meiotic inhibitor, for 24 h, then for an additional 16 h without TSA or hormones for a total of 40 h. At the end of maturation, oocytes were denuded and their zona pellucida’s removed. Oocytes were stained with anti-5-methylcytosine (5mC, 1:500). Fluorescent images of the oocytes were acquired, images were analyzed using ImageJ, and data analysis was performed using ANOVA and Tukey’s test. Oocytes matured with TSA had significantly greater (P < 0.05) levels of DNA methylation by the end of in vitro maturation compared to those not supplemented with TSA These results suggest that TSA can be used to develop an in vitro model to study the effects of epigenetic modifications in oocytes from aged livestock.

Gonadotropins-mediated bovine oocyte in vitro maturation and its genetic and epigenetic change- ART safety in vitro model

2013 ◽  
Vol 100 (3) ◽  
pp. S481
Author(s):  
C.-L. Lu ◽  
L.-Y. Yan ◽  
T.-R. Wang ◽  
R. Li ◽  
H.-L. Feng ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
In Vitro Model ◽  
Bovine Oocyte ◽  
Epigenetic Change ◽  
Vitro Model

scholarly journals Two-dimensional electrophoresis protein profiles of HL-60 and CCRF-CEM cell lines treated with epigenetic modification drugs

2019 ◽  
pp. 10-23
Author(s):  
Aziee Sudin ◽  
Haiyuni Mohd Yassim ◽  
Shafini Mohamed Yusoff ◽  
Shaharum Shamsuddin ◽  
Ridhwan Abdul Wahab ◽  
...  
Keyword(s):  
Cell Lines ◽  
Dna Methyltransferase ◽  
Epigenetic Modification ◽  
Trichostatin A ◽  
Lymphoblastic Leukemia ◽  
Genetic Alterations ◽  
Epigenetic Modifications ◽  
Hematopoietic Stem ◽  
2D Gel

Leukemia is classified as a malignant disease of hematopoietic stem cells (HSCs) that fails in cell differentiation but preserve their self-renewal. It is caused by genetic alterations and epigenetic modifications resulting in the activation or inactivation of particular genes for transcription. Epigenetic causes changes in gene expression without any alteration in the DNA sequence. The most common epigenetic modifications are DNA methylation and histone acetylation. 5-Azacitidine (5-Aza) is a DNA methytransferase inhibitor (DNMTi) that inhibits DNA methyltransferase enzymes resulting in hypomethylation. Trichostatin A (TSA) is a histone deacetylase inhibitor which inhibits deacetylation of both histone and non-histone proteins resulting in chromatin relaxation. This present study focused on the alteration of proteome profile on 2D gel electrophoresis (2-DE) induced by 5-Aza and TSA in HL-60 and CCRF-CEM cell lines as in vitro model to represent acute promyelocytic leukemia (APL) and T-lymphoblastic leukemia (T-ALL), respectively. Total proteins of untreated and 5-Aza/TSA-treated HL-60 and CCRF-CEM cell lines were extracted using urea/thiourea buffer and stained with Coomassie Blue. Comparative analysis of untreated and 5-Aza/TSA-treated HL-60 and CCRF-CEM was performed by PDQuest software. Qualitative analysis identified 190-659 protein spots detected in untreated, 5-Aza and TSA-treated HL-60 and CCRF-CEM. Quantitative comparison analysis was analyzed by over 2-fold change in 5-Aza/TSA-treated cells compared to untreated. One and eight upregulated proteins were detected in 5-Aza and TSA-treated HL-60, respectively. While five and one upregulated proteins were detected in 5-Aza and TSA-treated CCRF-CEM, respectively. These preliminary results suggested that 5-Aza and TSA induced proteome profiles alterations due to their inhibition effects in HL-60 and CCRF-CEM cell lines.

scholarly journals Testing of aerosolized ciprofloxacin nanocarriers on cystic fibrosis airway cells infected with P. aeruginosa biofilms

2021 ◽  
Author(s):  
Jenny Juntke ◽  
Xabier Murgia ◽  
Nazende Günday Türeli ◽  
Akif Emre Türeli ◽  
Chelsea R. Thorn ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
In Vitro Model ◽  
Time Window ◽  
Animal Experiments ◽  
Pulmonary Delivery ◽  
Barrier Properties ◽  
In Vitro Test ◽  
Safety And Efficacy ◽  
Vitro Model

AbstractThe major pathogen found in the lungs of adult cystic fibrosis (CF) patients is Pseudomonas aeruginosa, which builds antibiotic-resistant biofilms. Pulmonary delivery of antibiotics by inhalation has already been proved advantageous in the clinic, but the development of novel anti-infective aerosol medicines is complex and could benefit from adequate in vitro test systems. This work describes the first in vitro model of human bronchial epithelial cells cultivated at the air–liquid interface (ALI) and infected with P. aeruginosa biofilm and its application to demonstrate the safety and efficacy of aerosolized anti-infective nanocarriers. Such a model may facilitate the translation of novel therapeutic modalities into the clinic, reducing animal experiments and the associated problems of species differences. A preformed biofilm of P. aeruginosa PAO1 was transferred to filter-grown monolayers of the human CF cell line (CFBE41o-) at ALI and additionally supplemented with human tracheobronchial mucus. This experimental protocol provides an appropriate time window to deposit aerosolized ciprofloxacin-loaded nanocarriers at the ALI. When applied 1 h post-infection, the nanocarriers eradicated all planktonic bacteria and reduced the biofilm fraction of the pathogen by log 6, while CFBE41o- viability and barrier properties were maintained. The here described complex in vitro model approach may open new avenues for preclinical safety and efficacy testing of aerosol medicines against P. aeruginosa lung infection. Graphical abstract

scholarly journals Effects of microRNA-208a on inflammation and oxidative stress in ketamine-induced cardiotoxicity through Notch/NF-κB signal pathways by CHD9

2019 ◽  
Vol 39 (5) ◽  
Author(s):  
Hongjie Yuan ◽  
Shibin Du ◽  
Youliang Deng ◽  
Xiaoqing Xu ◽  
Qian Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
In Vitro Model ◽  
Epigenetic Modification ◽  
Heart Cell ◽  
Signal Pathways ◽  
Over Expression ◽  
Potential Biomarker ◽  
Vitro Model ◽  
And Oxidative Stress

Abstract Background: MicroRNA can regulate gene expression, and participate in multiple vital activities, such as inflammation, oxidative stress epigenetic modification, cell proliferation, and apoptosis. It plays an important role in the genesis and development of cardiovascular disease. Objective: To assess the role of microRNA-208a in ketamine-induced cardiotoxicity. Methods: All rats were randomly selected into two groups: sham and model groups. After fixed, all rats in the model group was intraperitoneally (IP) injected with 100 mg/kg of ketamine. Heart samples were stained with HE assay. Total RNAs from serum were used to hybridize with the SurePrint G3 Rat Whole Genome GE 8×60 K Microarray G4858A platform. Results: In the rat model with ketamine-induced cardiotoxicity, microRNA-208a expression was increased. Then, over-expression of microRNA-208a increased inflammation and oxidative stress in vitro model. However, down-regulation of microRNA-208a decreased inflammation and oxidative stress in vitro model. Over-expression of microRNA-208a suppressed CHD9 and Notch1, and induced p65 protein expression in vitro model. Overexpression of CHD9 reduced the effects of microRNA-208a on inflammation and oxidative stress in heart cell through Notch/p65 signal pathways. Notch1 activation reduced the effects of microRNA-208a on inflammation and oxidative stress in heart cell through p65 signal pathways. Conclusion: MicroRNA-208a may be a potential biomarker for ketamine-induced cardiotoxicity through inflammation and oxidative stress by Notch/NF-κB signal pathways by CHD9.

An in vitro model for investigation of vitamin A effects on wound healing

2021 ◽  
pp. 1-6
Author(s):  
Hoda Keshmiri Neghab ◽  
Mohammad Hasan Soheilifar ◽  
Gholamreza Esmaeeli Djavid
Keyword(s):  
Wound Healing ◽  
Endothelial Cell ◽  
Vitamin A ◽  
In Vitro Model ◽  
Cellular Proliferation ◽  
Endothelial Cell Migration ◽  
Normal Fibroblast ◽  
Anti Inflammatory ◽  
Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inflammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inflammation responses.

Sign in / Sign up

Export Citation Format

Share Document