scholarly journals Disruption of Tomato TGS Machinery by ToLCNDV Causes Reprogramming of Vascular Tissue Specific TORNADO1 Gene Expression

2021 ◽  
Author(s):  
Shreya Chowdhury ◽  
Shrabani Basak ◽  
Rohit Das ◽  
Arunava Mandal ◽  
Pallob Kundu
Keyword(s):  
Vascular Tissue ◽  
Leaf Growth ◽  
Methylation Status ◽  
Dna Methyltransferases ◽  
Dna Demethylation ◽  
Growth Stages ◽  
Developmental Defects ◽  
Leaf Venation ◽  
Leaf Curl

Abstract Leaf curl disease of tomato caused by Tomato Leaf Curl New Delhi Virus (ToLCNDV) inflicts huge economical loss. Disease symptoms resemble leaf developmental defects including abnormal vein architecture. Leaf vein patterning related TORNADO1 gene’s ( SlTRN1 ) transcript level is augmented in virus-infected leaves. To elucidate the molecular mechanism of the upregulation of SlTRN1 in vivo we have deployed SlTRN1 promoter-reporter transgenic tomato plants and investigated the gene’s dynamic expression pattern in leaf growth stages and infection. Expression of the gene was delimited in the vascular tissues and oppressed in growing leaves. Methylation-sensitive PCR analyses confirmed the accumulation of CHH methylation at multiple locations in the SlTRN1 promoter in older leaves. However, ToLCNDV infection reverses the methylation status and restores expression level in the leaf vascular bundle. The virus dampens the level of key maintenance and de novo DNA methyltransferases SlDRM5 , SlMET1 , SlCMT2 with concomitant augmentation of two DNA demethylases, SlDML1 and SlDML2 levels in SlTRN1 promoter-reporter transgenics. Transient overexpression of SlDML2 mimics the virus-induced hypomethylation state of the SlTRN1 promoter in mature leaves. Further, in line with the previous studies, we confirm the crucial role of viral suppressors of RNA silencing AC1 and AC4 proteins in promoting DNA demethylation and directing it to reinstate activated transcription of SlTRN1 in silenced tissues for possible modification of leaf venation architecture, leaf curling and easy vector acquisition of viral particles.

MO031KLOTHO GENE PROMOTER METHYLATION IN THE VASCULATURE IS RELATED TO INFLAMMATION IN HUMAN ATHEROSCLEROSIS

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Ernesto Martín-Núñez ◽  
Javier Donate-Correa ◽  
Carla Ferri ◽  
Víctor G Tagua ◽  
Ainhoa González-Luis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Promoter Region ◽  
Vascular Tissue ◽  
Methylation Status ◽  
Percent Level ◽  
Gene Promoter ◽  
Dna Methyltransferases ◽  
Case Group ◽  
Atherosclerotic Vascular Disease ◽  
Neutrophil Lymphocyte Ratio

Abstract Background and Aims Atherosclerosis, a vascular pathological process with an important inflammatory component, underlies most of the cardiovascular diseases (CVD). Deficiencies in the antiaging factor α-Klotho (KL) have been related to the appearance and progression of atherosclerotic damage associated with CVD. In addition, some studies have demonstrated that KL gene is expressed in human arteries and that low vascular KL expression levels are related with clinical atherosclerotic disease. Among the epigenetic mechanisms that regulate the expression of the KL gene, methylation of the CpG islands in the promoter region is one of the most studied. In this pilot study, we propose that this mechanism participates in the regulation of vascular KL expression and contributes to the deficiency in vascular KL levels observed in CVD. Moreover, we determined the influence of different inflammatory components in the methylation status of KL promoter. Method We developed a case-control study that included 25 patients with diagnosis of clinical atherosclerotic vascular disease undergoing elective vascular surgery and 15 cadaveric organ donors with no medical history of CVD, respectively. Vascular fragments were retrieved from all subjects, and the degree of methylation of the KL gene promoter was assessed by bisulfite sequencing. Additionally, vascular gene expression in both groups was assessed by qPCR for KL, two DNA methyltransferases (DNMT1 and DNMT3A), and three inflammatory-related loci (TNF, IL10 and NFKB1). Serum concentrations of TNFα and IL10 were measured by ELISA immunoassay, and complementary inflammatory parameters (CRP and neutrophil/lymphocyte ratio) by standardized routine methods. Results The vascular tissue of patients in the case group presented a higher percent level of methylated positions in the KL promoter region (71.4±8.3% vs. 39.4±12.3%, P<0.05), as well as a significant decrease in the expression of KL gene (a 40% reduction as compared to controls, P<0.05). Considering the whole study population, the methylation degree was inversely related to the endogenous transcript levels of KL gene (r=-0.66, P<0.0001). Moreover, the expression of DNMT1 was significantly increased in the case group (P<0.05), as well as those of the proinflammatory genes TNF and NFKB1 (P<0.0001 and P<0.05, respectively), being all of them directly associated with the vascular methylation levels of KL gene (r=0.51, P<0.0001 for DNMT1; r=0.45, P<0.001 for TNF; r= 0.37, P<0.05 for NFKB1). Furthermore, vascular expression of DNMT1 and DNMT3A directly correlated with both inflammatory markers in this tissue (P<0.0001, for all). Systemic levels of IL10 presented a direct association with vascular KL gene expression (r=0.28, P<0.05), while inverse associations were observed with the methylation degree of KL gene (r=-0.38, P<0.05) and DNMT1 expression (r=-0,29, P<0.05). Conclusion These results suggest that the methylation of KL gene promoter in the vasculature is a mechanism associated with the inflammatory response that would partly explain its reduction during atherosclerotic injury.

scholarly journals Genome-wide analysis of DNA methylation identifies the apoptosis-related gene UQCRH in renal cancer

2021 ◽  
Author(s):  
Kosuke Miyakuni ◽  
Jun Nishida ◽  
Daizo Koinuma ◽  
Genta Nagae ◽  
Hiroyuki Aburatani ◽  
...  
Keyword(s):  
Renal Cancer ◽  
Cancer Progression ◽  
Dna Methyltransferases ◽  
Tumor Formation ◽  
Dna Demethylation ◽  
Sequencing Data ◽  
Dna Hypermethylation ◽  
Cell Renal Cell Carcinoma ◽  
Methylation Array

Abstract Background DNA hypermethylation is frequently observed in clear cell renal cell carcinoma (ccRCC) and correlates with poor clinical outcomes. However, the detailed function is not fully uncovered. Methods A target for DNA methyltransferases (DNMTs) was explored in ccRCC cells. Highly malignant derivatives of human ccRCC cells were established via serial orthotopic inoculation in mice. Then, DNA methylated sites were genome-widely analyzed using methylation array in reference to RNA-sequencing data. Results We found that DNMT3B upregulation contributes to renal cancer progression and identified the ubiquinol cytochrome c reductase hinge protein (UQCRH) as a methylation target in advanced ccRCC. The expression of UQCRH in human ccRCC tissues was lower than that in normal adjacent tissues. Furthermore, its silencing attenuated cytochrome c release in response to apoptotic stimuli, resulting in enhanced primary tumor formation in vivo. Moreover, DNA demethylation enhanced the therapeutic efficiency of the mammalian target of rapamycin (mTOR) inhibitor everolimus in vivo. Conclusions These findings suggest that the DNMT3B-induced UQCRH methylation may contribute to renal cancer progression and that DNMT inhibitors exhibit potential clinical significance for ccRCC treatment.

Surface Modification by Nanobiomaterials for Vascular Tissue Engineering Applications

2020 ◽  
Vol 27 (10) ◽  
pp. 1634-1646 ◽  
Author(s):  
Huey-Shan Hung ◽  
Shan-hui Hsu
Keyword(s):  
Tissue Engineering ◽  
Vascular Tissue ◽  
Thrombus Formation ◽  
Vascular Grafts ◽  
Vascular Tissue Engineering ◽  
Great Success ◽  
Natural Polymers ◽  
Vascular Biomaterials

Treatment of cardiovascular disease has achieved great success using artificial implants, particularly synthetic-polymer made grafts. However, thrombus formation and restenosis are the current clinical problems need to be conquered. New biomaterials, modifying the surface of synthetic vascular grafts, have been created to improve long-term patency for the better hemocompatibility. The vascular biomaterials can be fabricated from synthetic or natural polymers for vascular tissue engineering. Stem cells can be seeded by different techniques into tissue-engineered vascular grafts in vitro and implanted in vivo to repair the vascular tissues. To overcome the thrombogenesis and promote the endothelialization effect, vascular biomaterials employing nanotopography are more bio-mimic to the native tissue made and have been engineered by various approaches such as prepared as a simple surface coating on the vascular biomaterials. It has now become an important and interesting field to find novel approaches to better endothelization of vascular biomaterials. In this article, we focus to review the techniques with better potential improving endothelization and summarize for vascular biomaterial application. This review article will enable the development of biomaterials with a high degree of originality, innovative research on novel techniques for surface fabrication for vascular biomaterials application.

scholarly journals Effective decellularisation of human saphenous veins for biocompatible arterial tissue engineering applications: Bench optimisation and feasibility in vivo testing

2021 ◽  
Vol 12 ◽  
pp. 204173142098752
Author(s):  
Nadiah S Sulaiman ◽  
Andrew R Bond ◽  
Vito D Bruno ◽  
John Joseph ◽  
Jason L Johnson ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Mechanical Strength ◽  
Vascular Tissue ◽  
Sodium Dodecyl ◽  
Host Cells ◽  
Replacement Model ◽  
In Vivo Testing ◽  
Vascular Scaffolds

Human saphenous vein (hSV) and synthetic grafts are commonly used conduits in vascular grafting, despite high failure rates. Decellularising hSVs (D-hSVs) to produce vascular scaffolds might be an effective alternative. We assessed the effectiveness of a detergent-based method using 0% to 1% sodium dodecyl sulphate (SDS) to decellularise hSV. Decellularisation effectiveness was measured in vitro by nuclear counting, DNA content, residual cell viability, extracellular matrix integrity and mechanical strength. Cytotoxicity was assessed on human and porcine cells. The most effective SDS concentration was used to prepare D-hSV grafts that underwent preliminary in vivo testing using a porcine carotid artery replacement model. Effective decellularisation was achieved with 0.01% SDS, and D-hSVs were biocompatible after seeding. In vivo xeno-transplantation confirmed excellent mechanical strength and biocompatibility with recruitment of host cells without mechanical failure, and a 50% patency rate at 4-weeks. We have developed a simple biocompatible methodology to effectively decellularise hSVs. This could enhance vascular tissue engineering toward future clinical applications.

scholarly journals Assessment of Electrospun Pellethane-Based Scaffolds for Vascular Tissue Engineering

Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3678
Author(s):  
Vera Chernonosova ◽  
Alexandr Gostev ◽  
Ivan Murashov ◽  
Boris Chelobanov ◽  
Andrey Karpenko ◽  
...  
Keyword(s):  
Vascular Tissue ◽  
Ex Vivo ◽  
Vascular Grafts ◽  
Wistar Rat ◽  
Graft Occlusion ◽  
Suitable Candidate ◽  
Large Animals ◽  
Cell Adhesion And Proliferation

We examined the physicochemical properties and the biocompatibility and hemocompatibility of electrospun 3D matrices produced using polyurethane Pellethane 2363-80A (Pel-80A) blends Pel-80A with gelatin or/and bivalirudin. Two layers of vascular grafts of 1.8 mm in diameter were manufactured and studied for hemocompatibility ex vivo and functioning in the infrarenal position of Wistar rat abdominal aorta in vivo (n = 18). Expanded polytetrafluoroethylene (ePTFE) vascular grafts of similar diameter were implanted as a control (n = 18). Scaffolds produced from Pel-80A with Gel showed high stiffness with a long proportional limit and limited influence of wetting on mechanical characteristics. The electrospun matrices with gelatin have moderate capacity to support cell adhesion and proliferation (~30–47%), whereas vascular grafts with bivalirudin in the inner layer have good hemocompatibility ex vivo. The introduction of bivalirudin into grafts inhibited platelet adhesion and does not lead to a change hemolysis and D-dimers concentration. Study in vivo indicates the advantages of Pel-80A grafts over ePTFE in terms of graft occlusion, calcification level, and blood velocity after 6 months of implantation. The thickness of neointima in Pel-80A–based grafts stabilizes after three months (41.84 ± 20.21 µm) and does not increase until six months, demonstrating potential for long-term functioning without stenosis and as a suitable candidate for subsequent preclinical studies in large animals.

scholarly journals In Vivo and In Vitro Assays Evaluating the Biological Activity of Taurine, Glucose and Energetic Beverages

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2198
Author(s):  
Marcos Mateo-Fernández ◽  
Fernando Valenzuela-Gómez ◽  
Rafael Font ◽  
Mercedes Del Río-Celestino ◽  
Tania Merinas-Amo ◽  
...  
Keyword(s):  
Dna Damage ◽  
Drosophila Melanogaster ◽  
Biological Activity ◽  
Biological Effects ◽  
Methylation Status ◽  
Repetitive Sequences ◽  
Energy Drinks ◽  
Red Bull

Taurine is one of the main ingredients used in energy drinks which are highly consumed in adolescents for their sugary taste and stimulating effect. With energy drinks becoming a worldwide phenomenon, the biological effects of these beverages must be evaluated in order to fully comprehend the potential impact of these products on the health due to the fact nutrition is closely related to science since the population consumes food to prevent certain diseases. Therefore, the aim of this study was to evaluate the biological effects of taurine, glucose, classic Red Bull® and sugar-free Red Bull® in order to check the food safety and the nutraceutical potential of these compounds, characterising different endpoints: (i) Toxicology, antitoxicology, genotoxicology and life expectancy assays were performed in the Drosophila melanogaster model organism; (ii) The in vitro chemopreventive activity of testing compounds was determined by assessing their cytotoxicity, the proapoptotic DNA-damage capability to induce internucleosomal fragmentation, the strand breaks activity and the modulator role on the methylation status of genomic repetitive sequences of HL-60 promyelocytic cells. Whereas none tested compounds showed toxic or genotoxic effect, all tested compounds exerted antitoxic and antigenotoxic activity in Drosophila. Glucose, classic Red Bull® and sugar-free Red Bull® were cytotoxic in HL-60 cell line. Classic Red Bull® induced DNA internucleosomal fragmentation although none of them exhibited DNA damage on human leukaemia cells. In conclusion, the tested compounds are safe on Drosophila melanogaster and classic Red Bull® could overall possess nutraceutical potential in the in vivo and in vitro model used in this study. Besides, taurine could holistically be one of the bioactive compounds responsible for the biological activity of classic Red Bull®.

scholarly journals Integrated Physiological and Transcriptomic Analyses Reveal a Regulatory Network of Anthocyanin Metabolism Contributing to the Ornamental Value in a Novel Hybrid Cultivar of Camellia japonica

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1724
Author(s):  
Liqin Pan ◽  
Jiyuan Li ◽  
Hengfu Yin ◽  
Zhengqi Fan ◽  
Xinlei Li
Keyword(s):  
Regulatory Network ◽  
Leaf Growth ◽  
Expression Patterns ◽  
Differential Expression Analysis ◽  
Growth Stages ◽  
Potential Candidate ◽  
Leaf Color ◽  
Camellia Japonica ◽  
Leaf Phenotype ◽  
Hybrid Cultivar

Camellia japonica is a plant species with great ornamental and gardening values. A novel hybrid cultivar Chunjiang Hongxia (Camellia japonica cv. Chunjiang Hongxia, CH) possesses vivid red leaves from an early growth stage to a prolonged period and is, therefore, commercially valuable. The molecular mechanism underlying this red-leaf phenotype in C. japonica cv. CH is largely unknown. Here, we investigated the leaf coloration process, photosynthetic pigments contents, and different types of anthocyanin compounds in three growth stages of the hybrid cultivar CH and its parental cultivars. The gene co-expression network and differential expression analysis from the transcriptome data indicated that the changes of leaf color were strongly correlated to the anthocyanin metabolic processes in different leaf growth stages. Genes with expression patterns associated with leaf color changes were also discussed. Together, physiological and transcriptomic analyses uncovered the regulatory network of metabolism processes involved in the modulation of the ornamentally valuable red-leaf phenotype and provided the potential candidate genes for future molecular breeding of ornamental plants such as Camellia japonica.

Methylation status of oestrogen receptor α-A: a predictor of prognosis in leukaemias

2010 ◽  
Vol 30 (4) ◽  
pp. 217-222 ◽  
Author(s):  
Jie Yao ◽  
Xiao-Bing Zhang ◽  
Xiao-li Zhang ◽  
Wei-Ling Fu
Keyword(s):  
Protein Expression ◽  
Cell Apoptosis ◽  
Thymidine Incorporation ◽  
Symptomatic Relief ◽  
Methylation Status ◽  
Dna Demethylation ◽  
Blue Staining ◽  
Chemotherapy Treatment ◽  
Propidium Iodide Staining ◽  
Epigenetic Biomarker

Many studies have shown that epigenetic regulation of ERs (oestrogen receptors) plays a key role in the pathogenesis of leukaemia. In the present study, it was found that the methylated status of ERα-A might serve as an epigenetic biomarker of leukaemias. In this study, the protein expression and cell apoptosis, cycle, proliferation and viability with and without 5-aza-dC (5-aza-2′-deoxycytidine) were evaluated with Western blotting, 3H-TdR (3H-thymidine) incorporation, propidium iodide staining and Trypan Blue staining respectively. The protein expression of ERα was significantly enhanced in all leukaemic cell lines using treatment with the DNA demethylation reagent 5-aza-dC. However, no obvious change in the protein expression of ERβ takes place with 5-aza-dC. And with 5-aza-dC, cell apoptosis, cell cycle, cell proliferation and viability were all inhibited significantly. We also tracked 40 cases of leukaemias with ERα-A methylation (95%; 38 of 40) to observe the prognosis 1 year after chemotherapy treatment. The patients with ERα-A methylation have no obvious symptomatic relief; however, two patients without ERα-A methylation have obtained effective relief. This result suggested that ERα plays a significant role in leukaemogenesis, and the methylated status of ERα-A not only might serve as an epigenetic biomarker of leukaemias for diagnosis, but also has the potential to serve as a predictor of prognosis in leukaemias.

The growth and development of the leaf in tomato (Lycopersicon esculentum). I. The plastochron index, a suitable basis for description

1976 ◽  
Vol 54 (21) ◽  
pp. 2421-2428 ◽  
Author(s):  
Warren K. Coleman ◽  
Richard I. Greyson
Keyword(s):  
Lycopersicon Esculentum ◽  
Seasonal Variability ◽  
Growth And Development ◽  
Growth Analysis ◽  
Growth Habit ◽  
Leaf Growth ◽  
Growth Stages ◽  
Flower Buds ◽  
Morphological Status ◽  
Plastochron Index

Growth analysis indicates that the plastochron index (PI) is consistently applicable for describing the morphological status of the vegetative tomato shoot in quantitative terms, and pronounced seasonal variability in the growth habit is minimized. However, the PI is not applicable to tomato shoots after flower buds are produced at the vegetative apex. True leaf no. 3 goes through four growth stages which can be characterized in terms of the leaf plastochron index (LPI3). A basipetal trend in leaf growth and development is evident.

scholarly journals Polyamine Metabolism and Gene Methylation in Conjunction with One-Carbon Metabolism

2018 ◽  
Vol 19 (10) ◽  
pp. 3106 ◽  
Author(s):  
Kuniyasu Soda
Keyword(s):  
Dna Methylation ◽  
Carbon Metabolism ◽  
Methylation Status ◽  
Significant Risk ◽  
Significant Risk Factor ◽  
Dna Methyltransferases ◽  
Polyamine Metabolism ◽  
Methyl Group ◽  
Wide Range ◽  
One Carbon Metabolism

Recent investigations have revealed that changes in DNA methylation status play an important role in aging-associated pathologies and lifespan. The methylation of DNA is regulated by DNA methyltransferases (DNMT1, DNMT3a, and DNMT3b) in the presence of S-adenosylmethionine (SAM), which serves as a methyl group donor. Increased availability of SAM enhances DNMT activity, while its metabolites, S-adenosyl-l-homocysteine (SAH) and decarboxylated S-adenosylmethionine (dcSAM), act to inhibit DNMT activity. SAH, which is converted from SAM by adding a methyl group to cytosine residues in DNA, is an intermediate precursor of homocysteine. dcSAM, converted from SAM by the enzymatic activity of adenosylmethionine decarboxylase, provides an aminopropyl group to synthesize the polyamines spermine and spermidine. Increased homocysteine levels are a significant risk factor for the development of a wide range of conditions, including cardiovascular diseases. However, successful homocysteine-lowering treatment by vitamins (B6, B12, and folate) failed to improve these conditions. Long-term increased polyamine intake elevated blood spermine levels and inhibited aging-associated pathologies in mice and humans. Spermine reversed changes (increased dcSAM, decreased DNMT activity, aberrant DNA methylation, and proinflammatory status) induced by the inhibition of ornithine decarboxylase. The relation between polyamine metabolism, one-carbon metabolism, DNA methylation, and the biological mechanism of spermine-induced lifespan extension is discussed.

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