scholarly journals Redox-Dependent Structural Modification of Nucleoredoxin Triggers Defense Responses against Alternaria brassicicola in Arabidopsis

2020 ◽  
Vol 21 (23) ◽  
pp. 9196
Author(s):  
Chang Ho Kang ◽  
Joung Hun Park ◽  
Eun Seon Lee ◽  
Seol Ki Paeng ◽  
Ho Byoung Chae ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Structural Modification ◽  
Quantitative Polymerase Chain Reaction ◽  
Defense Responses ◽  
Alternaria Brassicicola ◽  
Low Molecular Weight ◽  
Biological Processes ◽  
Wild Type ◽  
Molecular Sensor ◽  
Polymerase Chain

In plants, thioredoxin (TRX) family proteins participate in various biological processes by regulating the oxidative stress response. However, their role in phytohormone signaling remains largely unknown. In this study, we investigated the functions of TRX proteins in Arabidopsis thaliana. Quantitative polymerase chain reaction (qPCR) experiments revealed that the expression of ARABIDOPSIS NUCLEOREDOXIN 1 (AtNRX1) is specifically induced by the application of jasmonic acid (JA) and upon inoculation with a necrotrophic fungal pathogen, Alternaria brassicicola. The AtNRX1 protein usually exists as a low molecular weight (LMW) monomer and functions as a reductase, but under oxidative stress AtNRX1 transforms into polymeric forms. However, the AtNRX1M3 mutant protein, harboring four cysteine-to-serine substitutions in the TRX domain, did not show structural modification under oxidative stress. The Arabidopsisatnrx1 null mutant showed greater resistance to A. brassicicola than wild-type plants. In addition, plants overexpressing both AtNRX1 and AtNRX1M3 were susceptible to A. brassicicola infection. Together, these findings suggest that AtNRX1 normally suppresses the expression of defense-responsive genes, as if it were a safety pin, but functions as a molecular sensor through its redox-dependent structural modification to induce disease resistance in plants.

scholarly journals Andrographis paniculata and Its Bioactive Diterpenoids Against Inflammation and Oxidative Stress in Keratinocytes

Antioxidants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 530 ◽  
Author(s):  
Eugenie Mussard ◽  
Sundy Jousselin ◽  
Annabelle Cesaro ◽  
Brigitte Legrain ◽  
Eric Lespessailles ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Quantitative Polymerase Chain Reaction ◽  
Andrographis Paniculata ◽  
Ros Production ◽  
Inflammatory Conditions ◽  
Tnf Α ◽  
Dichlorofluorescein Diacetate ◽  
Polymerase Chain ◽  
Factor Α ◽  
Necrosis Factor

Andrographis paniculata was widely used in traditional herbal medicine to treat various diseases. This study explored the potential anti-aging activity of Andrographis paniculata in cutaneous cells. Human, adult, low calcium, high temperature (HaCaT) cells were treated with methanolic extract (ME), andrographolide (ANDRO), neoandrographolide (NEO), 14-deoxyandrographolide (14DAP) and 14-deoxy-11,12-didehydroandrographolide (14DAP11-12). Oxidative stress and inflammation were induced by hydrogen peroxide and lipopolysaccharide/TNF-α, respectively. Reactive oxygen species (ROS) production was measured by fluorescence using a 2′,7′-dichlorofluorescein diacetate (DCFH-DA) probe and cytokines were quantified by ELISA for interleukin-8 (IL-8) or reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for tumor necrosis factor-α (TNF-α). Hyaluronic acid (HA) secretion was determined by an ELISA. Our results show a decrease in ROS production and TNF-α expression by ME (5 µg/mL) in HaCaT under pro-oxidant and pro-inflammatory conditions, respectively. ME protected HaCaT against oxidative stress and inflammation. Our findings confirm that ME can be used for the development of bioactive compounds against epidermal damage.

scholarly journals Evaluation of toxicity and response to oxidative stress generated by orthodontic bands in human gingival fibroblasts

2019 ◽  
Vol 90 (2) ◽  
pp. 285-290 ◽  
Author(s):  
Alexandre Marcos Bandeira ◽  
Elizabeth Ferreira Martinez ◽  
Ana Paula Dias Demasi
Keyword(s):  
Oxidative Stress ◽  
Quantitative Polymerase Chain Reaction ◽  
Conditioned Media ◽  
Human Gingival Fibroblasts ◽  
Gingival Fibroblasts ◽  
Trypan Blue Exclusion ◽  
Antioxidant Genes ◽  
Glutathione Peroxidase 1 ◽  
Viable Cells ◽  
Polymerase Chain

ABSTRACT Objective: To evaluate the cytotoxicity of stainless-steel orthodontic bands and their influence on the expression of the antioxidant genes in human gingival fibroblasts. Materials and Methods: Ten bands of each brand (Dentsply-Sirona, Dentaurum, TP Orthodontics, and Morelli) were conditioned in 0.2 g/mL culture medium at 37°C for 14 days, and the corresponding conditioned media were applied over the fibroblasts. Cell viability was assessed after 24, 48, and 72 hours of exposure to the conditioned media by trypan blue exclusion assay. Expression of the antioxidant defense genes peroxiredoxin 1 (PRDX1), superoxide dismutase 1 (SOD1), and glutathione peroxidase 1 (GPX1) were evaluated by quantitative polymerase chain reaction after 24 hours of exposure. These parameters were compared to those of the cells not exposed to the conditioned media of the bands (control). Results: All bands promoted a reduction in the number of viable cells in the periods of 48 and 72 hours (P < .01). Analysis of gene expression showed a significant increase in the levels of PRDX1 transcripts caused by the conditioned media of the Dentsply-Sirona, TP Orthodontics, and Morelli bands (P < .01) as well as induction of SOD1 by the conditioned media of the Dentaurum and Morelli (P < .01). Expression of GPX1 was not influenced by the conditioned media. Conclusions: The orthodontic bands showed toxicity to fibroblasts and increased the expression of PRDX1 and SOD1 antioxidant genes, indicating induction of oxidative stress in the cells.

scholarly journals Controlled Human Infection With Bordetella pertussis Induces Asymptomatic, Immunizing Colonization

2019 ◽  
Vol 71 (2) ◽  
pp. 403-411 ◽  
Author(s):  
Hans de Graaf ◽  
Muktar Ibrahim ◽  
Alison R Hill ◽  
Diane Gbesemete ◽  
Andrew T Vaughan ◽  
...  
Keyword(s):  
Bordetella Pertussis ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Infection ◽  
Wild Type ◽  
Immunological Responses ◽  
Inoculum Dose ◽  
Colony Forming Units ◽  
Community Transmission ◽  
Polymerase Chain ◽  
Immunoglobin G

Abstract Background Bordetella pertussis is among the leading causes of vaccine-preventable deaths and morbidity globally. Human asymptomatic carriage as a reservoir for community transmission of infections might be a target of future vaccine strategies, but has not been demonstrated. Our objective was to demonstrate that asymptomatic nasopharyngeal carriage of Bordetella pertussis is inducible in humans and to define the microbiological and immunological features of presymptomatic infection. Methods Healthy subjects aged 18–45 years with an antipertussis toxin immunoglobin G (IgG) concentration of <20 international units/ml were inoculated intranasally with nonattenuated, wild-type Bordetella pertussis strain B1917. Safety, colonization, and shedding were monitored over 17 days in an inpatient facility. Colonization was assessed by culture and quantitative polymerase chain reaction. Azithromycin was administered from Day 14. The inoculum dose was escalated, aiming to colonize at least 70% of participants. Immunological responses were measured. Results There were 34 participants challenged, in groups of 4 or 5. The dose was gradually escalated from 103 colony-forming units (0% colonized) to 105 colony-forming units (80% colonized). Minor symptoms were reported in a minority of participants. Azithromycin eradicated colonization in 48 hours in 88% of colonized individuals. Antipertussis toxin IgG seroconversion occurred in 9 out of 19 colonized participants and in none of the participants who were not colonized. Nasal wash was a more sensitive method to detect colonization than pernasal swabs. No shedding of Bordetella pertussis was detected in systematically collected environmental samples. Conclusions Bordetella pertussis colonization can be deliberately induced and leads to a systemic immune response without causing pertussis symptoms. Clinical Trials Registration NCT03751514.

scholarly journals Expression of Putative Defense Responses in Cannabis Primed by Pseudomonas and/or Bacillus Strains and Infected by Botrytis cinerea

2020 ◽  
Vol 11 ◽  
Author(s):  
Carole Balthazar ◽  
Gabrielle Cantin ◽  
Amy Novinscak ◽  
David L. Joly ◽  
Martin Filion
Keyword(s):  
Botrytis Cinerea ◽  
Cannabis Sativa ◽  
Molecular Level ◽  
Quantitative Polymerase Chain Reaction ◽  
Defense Responses ◽  
Gray Mold ◽  
Control Measures ◽  
Effective Control ◽  
Defense Genes ◽  
Polymerase Chain

Cannabis (Cannabis sativa L.) offers many industrial, agricultural, and medicinal applications, but is commonly threatened by the gray mold disease caused by the fungus Botrytis cinerea. With few effective control measures currently available, the use of beneficial rhizobacteria represents a promising biocontrol avenue for cannabis. To counter disease development, plants rely on a complex network of inducible defense pathways, allowing them to respond locally and systemically to pathogens attacks. In this study, we present the first attempt to control gray mold in cannabis using beneficial rhizobacteria, and the first investigation of cannabis defense responses at the molecular level. Four promising Pseudomonas (LBUM223 and WCS417r) and Bacillus strains (LBUM279 and LBUM979) were applied as single or combined root treatments to cannabis seedlings, which were subsequently infected by B. cinerea. Symptoms were recorded and the expression of eight putative defense genes was monitored in leaves by reverse transcription quantitative polymerase chain reaction. The rhizobacteria did not significantly control gray mold and all infected leaves were necrotic after a week, regardless of the treatment. Similarly, no systemic activation of putative cannabis defense genes was reported, neither triggered by the pathogen nor by the rhizobacteria. However, this work identified five putative defense genes (ERF1, HEL, PAL, PR1, and PR2) that were strongly and sustainably induced locally at B. cinerea’s infection sites, as well as two stably expressed reference genes (TIP41 and APT1) in cannabis. These markers will be useful in future researches exploring cannabis defense pathways.

scholarly journals Ergothioneine Is a Secreted Antioxidant in Mycobacterium smegmatis

2013 ◽  
Vol 57 (7) ◽  
pp. 3202-3207 ◽  
Author(s):  
Carine Sao Emani ◽  
Monique J. Williams ◽  
Ian J. Wiid ◽  
Nicholas F. Hiten ◽  
Albertus J. Viljoen ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Mycobacterium Smegmatis ◽  
Double Mutant ◽  
Culture Media ◽  
Growth Cycle ◽  
Low Molecular Weight ◽  
Deficient Mutant ◽  
Wild Type ◽  
Content Type

ABSTRACTErgothioneine (ERG) and mycothiol (MSH) are two low-molecular-weight thiols synthesized by mycobacteria. The role of MSH has been extensively investigated in mycobacteria; however, little is known about the role of ERG in mycobacterial physiology. In this study, quantification of ERG at various points in the growth cycle ofMycobacterium smegmatisrevealed that a significant portion of ERG is found in the culture media, suggesting that it is actively secreted. A mutant ofM. smegmatislackingegtD(MSMEG_6247) was unable to synthesize ERG, confirming its role in ERG biosynthesis. Deletion ofegtDfrom wild-typeM. smegmatisand an MSH-deficient mutant did not affect their susceptibility to antibiotics tested in this study. The ERG- and MSH-deficient double mutant was significantly more sensitive to peroxide than either of the single mutants lacking either ERG or MSH, suggesting that both thiols play a role in protectingM. smegmatisagainst oxidative stress and that ERG is able to partly compensate for the loss of MSH.

scholarly journals Dimethyl itaconate alleviates the pyroptosis of macrophages through oxidative stress

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Shan-Shan Huang ◽  
Dong-Yang Guo ◽  
Bing-Bing Jia ◽  
Guo-Long Cai ◽  
Jing Yan ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Quantitative Polymerase Chain Reaction ◽  
Enrichment Analysis ◽  
Go Enrichment ◽  
Polymerase Chain ◽  
Stress Related Genes ◽  
Go Enrichment Analysis ◽  
Necrosis Factor ◽  
Differential Genes ◽  
Dimethyl Itaconate

AbstractMacrophages are involved in the pathophysiology of many diseases as critical cells of the innate immune system. Pyroptosis is a form of macrophage death that induces cytokinesis of phagocytic substances in the macrophages, thereby defending against infection. Dimethyl itaconate (DI) is an analog of itaconic acid with anti-inflammatory effects. However, the effect of dimethyl itaconate on macrophage pyroptosis has not been elucidated clearly. Thus, the present study aimed to analyze the effect of DI treatment on a macrophage pyroptosis model (Lipopolysaccharide, LPS + Adenosine Triphosphate, ATP). The results showed that 0.25 mM DI ameliorated macrophage pyroptosis and downregulated interleukin (IL)-1β expression. Then, real-time quantitative polymerase chain reaction (RT-qPCR) was used to confirm the result of RNA-sequencing of the upregulated oxidative stress-related genes (Gclc and Gss) and downregulated inflammation-related genes (IL-12β and IL-1β). In addition, Gene Ontology (GO) enrichment analysis showed that differential genes were associated with transcript levels and DNA replication. Kyoto encyclopedia of genes and genomes (KEGG) enrichment showed that signaling pathways, such as tumor necrosis factor (TNF), Jak, Toll-like receptor and IL-17, were altered after DI treatment. N-acetyl-L-cysteine (NAC) reversed the DI effect on the LPS + ATP-induced macrophage pyroptosis and upregulated the IL-1β expression. Oxidative stress-related protein Nrf2 is involved in the DI regulation of macrophage pyroptosis. Taken together, these findings suggested that DI alleviates the pyroptosis of macrophages through oxidative stress.

scholarly journals Tissue-Specific Profiling of Oxidative Stress-Associated Transcriptome in a Healthy Mouse Model

2018 ◽  
Vol 19 (10) ◽  
pp. 3174 ◽  
Author(s):  
Jung Kim ◽  
Hyeong Kim ◽  
Chang Son
Keyword(s):  
Oxidative Stress ◽  
Quantitative Polymerase Chain Reaction ◽  
Specific Gene ◽  
Tissue Specific ◽  
Age Related ◽  
Wide Range ◽  
Liver And Kidney ◽  
Specific Variation ◽  
Polymerase Chain ◽  
Lung And Kidney

Oxidative stress is a common phenomenon and is linked to a wide range of diseases and pathological processes including aging. Tissue-specific variation in redox signaling and cellular responses to oxidative stress may be associated with vulnerability especially to age-related and chronic diseases. In order to provide a basis for tissue-specific difference, we examined the tissue-specific transcriptional features of 101 oxidative stress-associated genes in 10 different tissues and organs of healthy mice under physiological conditions. Microarray analysis results, which were consistent with quantitative polymerase chain reaction (qPCR) results, showed that catalase, Gpx3, and Gpx4 were most highly regulated in the liver, kidney, and testes. We also found the tissue-specific gene expression of SOD1 (liver and kidney), SOD2 (heart and muscle), and SOD3 (lung and kidney). The current results will serve as a reference for animal models and help advance our understanding of tissue-specific variability in oxidative stress-associated pathogenesis.

scholarly journals Identification of 118 Arabidopsis Transcription Factor and 30 Ubiquitin-Ligase Genes Responding to Chitin, a Plant-Defense Elicitor

2007 ◽  
Vol 20 (8) ◽  
pp. 900-911 ◽  
Author(s):  
Marc Libault ◽  
Jinrong Wan ◽  
Tomasz Czechowski ◽  
Michael Udvardi ◽  
Gary Stacey
Keyword(s):  
Transcription Factor ◽  
Plant Defense ◽  
Ubiquitin Ligase ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Defense Responses ◽  
Plant Defense Responses ◽  
Genome Array ◽  
Polymerase Chain ◽  
Specific Regulation

Chitin, found in the cell walls of true fungi and the exoskeleton of insects and nematodes, is a well-established elicitor of plant defense responses. In this study, we analyzed the expression patterns of Arabidopsis thaliana transcription factor (TF) and ubiquitin-ligase genes in response to purified chitooctaose at different treatment times (15, 30, 60, 90, and 120 min after treatment), using both quantitative polymerase chain reaction and the Affymetrix Arabidopsis whole-genome array. A total of 118 TF genes and 30 ubiquitin-ligase genes were responsive to the chitin treatment. Among these genes, members from the following four TF families were overrepresented: APETALA2/ethylene-reponsive element binding proteins (27), C2H2 zinc finger proteins (14), MYB domain-containing proteins (11), and WRKY domain transcription factors (14). Transcript variants from a few of these genes were found to respond differentially to chitin, suggesting transcript-specific regulation of these TF genes.

scholarly journals Laser Irradiation Alters the Expression Profile of Genes Involved in the Extracellular MatrixIn Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-17 ◽  
Author(s):  
Sandra M. Ayuk ◽  
Nicolette N. Houreld ◽  
Heidi Abrahamse
Keyword(s):  
Laser Irradiation ◽  
Continuous Wave ◽  
Quantitative Polymerase Chain Reaction ◽  
Extracellular Proteins ◽  
Biological Processes ◽  
Cell Models ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Intensity Laser

The extracellular matrix (ECM) forms the basis of every phase in wound healing. Healing may be impaired if some of these components are destroyed. Photobiostimulation has demonstrated a stimulatory response in biological processes. This study aimed to evaluate various genes involved in the ECM, in response to laser irradiation. Isolated human skin fibroblasts were used in three different cell models, namely, normal, normal wounded, and diabetic wounded. Cells were irradiated with 5 J/cm2using a continuous wave diode laser emitting at a wavelength of 660 nm and incubated for 48 h. Nonirradiated (0 J/cm2) normal and diabetic wounded cells served as the control. Real-time reverse transcription (RT) quantitative polymerase chain reaction (qPCR) was used to determine the expression of 84 genes in a PCR array. There was a significant upregulation of 29 genes in the normal cells, 32 genes in the normal wounded cells, and 18 genes in the diabetic wounded cells as well as a downregulation of 19 genes (normal), 6 genes (normal wounded), and 31 genes (diabetic wounded). Low intensity laser irradiation (LILI) stimulates gene expression in various cell adhesion molecules (CAMs) and extracellular proteins at 660 nm in wounded fibroblastsin vitro.

scholarly journals Identification of Gene Candidates Associated with Huanglongbing Tolerance, Using ‘Candidatus Liberibacter asiaticus’ Flagellin 22 as a Proxy to Challenge Citrus

2018 ◽  
Vol 31 (2) ◽  
pp. 200-211 ◽  
Author(s):  
Qingchun Shi ◽  
Vicente J. Febres ◽  
Shujian Zhang ◽  
Fahong Yu ◽  
Greg McCollum ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Citrus Canker ◽  
Defense Responses ◽  
Candidatus Liberibacter Asiaticus ◽  
Tolerance Mechanisms ◽  
Molecular Pattern ◽  
Number Of Genes ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Polymerase Chain

The 22–amino acid (flg22) pathogen-associated molecular pattern from the flagellin of Xanthomonas citri subsp. citri has been shown to induce defense responses correlated with citrus canker resistance. Here, flg22 of ‘Candidatus Liberibacter asiaticus’, the putative causal agent of Huanglongbing (HLB), elicited differential defense responses that were weaker than those from Xcc-flg22, between those of the HLB-tolerant mandarin cultivar Sun Chu Sha and susceptible grapefruit cultivar Duncan. Transcriptomics was used to compare the effect of CLas-flg22 and Xcc-flg22 between the citrus genotypes and identified 86 genes induced only by CLas-flg22 in the tolerant mandarin. Expression of 16 selected genes was validated, by reverse transcription-quantitative polymerase chain reaction, and was evaluated in citrus during ‘Ca. L. asiaticus’ infection. Differential expression of a number of genes occurred between tolerant and susceptible citrus infected with ‘Ca. L. asiaticus’, suggesting their involvement in HLB tolerance. In addition, several genes were similarly regulated by CLas-flg22 and ‘Ca. L. asiaticus’ treatments, while others were oppositely regulated in the tolerant mandarin, suggesting similarity and interplay between CLas-flg22 and ‘Ca. L. asiaticus’–triggered defenses. Genes identified are valuable in furthering the study of HLB tolerance mechanisms and, potentially, for screening for HLB-tolerant citrus using CLas-flg22 as a pathogen proxy.

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