scholarly journals Gene Expression of Putative Pathogenicity-Related Genes in Verticillium dahliae in Response to Elicitation with Potato Extracts and during Infection Using Quantitative Real-Time PCR

Pathogens ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 510
Author(s):  
Xiaohan Zhu ◽  
Arbia Arfaoui ◽  
Mohammad Sayari ◽  
Lorne R. Adam ◽  
Fouad Daayf
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Functional Characterization ◽  
Regulatory Protein ◽  
Phosphorus Acquisition ◽  
Quantitative Real Time Pcr ◽  
Potato Leaf ◽  
Nadh Ubiquinone Oxidoreductase ◽  
Ubiquitin Conjugating Enzyme ◽  
New Ideas

Quantitative real-time PCR was used to monitor the expression of 15 Verticillium dahliae’s genes, putatively involved in pathogenicity, highly (HAV) and weakly aggressive (WAV) V. dahliae isolates after either (i) elicitation with potato leaf, stem, or root extracts, or (ii) inoculation of potato detached petioles. These genes, i.e., coding for Ras-GAP-like protein, serine/threonine protein kinase, Ubiquitin-conjugating enzyme variant-MMS2, NADH-ubiquinone oxidoreductase, Thioredoxin, Pyruvate dehydrogenase E1 VdPDHB, myo-inositol 2-dehydrogenase, and HAD-superfamily hydrolase, showed differential upregulation in the HAV versus WAV isolate in response to plant extracts or after inoculation of potato leaf petioles. This suggests their potential involvement in the observed differential aggressiveness between isolates. However, other genes like glucan endo-1,3-alpha-glucosidase and nuc-1 negative regulatory protein VdPREG showed higher activity in the WAV than in the HAV in response to potato extracts and/or during infection. This, in contrast, may suggest a role in their lower aggressiveness. These findings, along with future functional analysis of selected genes, will contribute to improving our understanding of V. dahliae’s pathogenesis. For example, expression of VdPREG negatively regulates phosphorus-acquisition enzymes, which may indicate a lower phosphorus acquisition activity in the WAV. Therefore, integrating the knowledge about the activity of both genes enhancing pathogenicity and those restraining it will provide a guild line for further functional characterization of the most critical genes, thus driving new ideas towards better Verticillium wilt management.

scholarly journals ABCC5 Gene Variant c.1146A>G Reduces MRP5 Expression and Can be a Potential Marker to Manage Cilostazol Induced Headache

2020 ◽  
Author(s):  
Dong-Hwan Lee ◽  
Woo-Dae Bang ◽  
Md. Hasanuzzaman ◽  
So Myoung Kim ◽  
Dong-Jin Kim ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Gene Transfection ◽  
Regulatory Protein ◽  
Genetic Variations ◽  
Severe Form ◽  
Intracellular Camp ◽  
Quantitative Real Time Pcr ◽  
Protective Allele ◽  
Potential Marker

Abstract Background: Patients taking cilostazol, a representative phosphodiesterase type III inhibitor used for vasodilation, occasionally complain of headaches. Cerebral arteriolar relaxation, due to an increase in intracellular cAMP or cGMP, is believed to be associated with a severe form of cilostazol-induced headaches. Multidrug resistance protein 5 (MRP5) is an important regulator of cAMP and cGMP could be a regulatory protein of this cilostazol induced headache.Methods: The response to cilostazol on the basis of MRP5 genetic variations was studied in phase-I clinical trial including 101 healthy Korean individuals. Quantitative real time PCR, Western blot, confocal analysis and drug transporter assay was performed to detect and evaluate the activity of MRP5. Statistical analysis was performed by using SPSS 18.0 and GraphPad Prism 4.0 software. Results: Population genetic and pharmacokinetic analyses indicated that a group of MRP5 genetic variations in linkage disequilibrium with c.1146A>G (rs7636910) is associated with the no or mild forms of cilostazol-induced headaches, but did not affect the systemic distribution of cilostazol or its metabolites. Quantitative real-time PCR and pyrosequencing assays of blood cells revealed that individuals with the c.1146G allele, a protective allele against cilostazol-induced headaches, had a 0.68-fold lower mRNA expression of MRP5 than that of individuals with c.1146A allele. In addition, in a gene transfection experiment using MDCKII cells, the G variant was found to be reduced the mRNA and protein expression of MRP5.Conclusion: These results suggest that MRP5 has an important role in the regulation of cilostazol-induced chronic headache and the c.1146A>G variation could be a potential marker to manage/treat cilostazol-induced headaches.Trial registration: The trial is registered at www.clinicaltrials.gov on (October 20, 2011) and the number is NCT01455558.

scholarly journals KLK11 promotes the activation of mTOR and protein synthesis to facilitate cardiac hypertrophy

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yi Wang ◽  
Hongjuan Liao ◽  
Yueheng Wang ◽  
Jinlin Zhou ◽  
Feng Wang ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Angiotensin Ii ◽  
Cardiac Hypertrophy ◽  
Western Blot ◽  
Real Time ◽  
Real Time Pcr ◽  
Mtor Signaling ◽  
Heart Weight ◽  
Cardiomyocyte Hypertrophy ◽  
Quantitative Real Time Pcr

Abstract Background Cardiovascular diseases have become the leading cause of death worldwide, and cardiac hypertrophy is the core mechanism underlying cardiac defect and heart failure. However, the underlying mechanisms of cardiac hypertrophy are not fully understood. Here we investigated the roles of Kallikrein 11 (KLK11) in cardiac hypertrophy. Methods Human and mouse hypertrophic heart tissues were used to determine the expression of KLK11 with quantitative real-time PCR and western blot. Mouse cardiac hypertrophy was induced by transverse aortic constriction (TAC), and cardiomyocyte hypertrophy was induced by angiotensin II. Cardiac function was analyzed by echocardiography. The signaling pathway was analyzed by western blot. Protein synthesis was monitored by the incorporation of [3H]-leucine. Gene expression was analyzed by quantitative real-time PCR. Results The mRNA and protein levels of KLK11 were upregulated in human hypertrophic hearts. We also induced cardiac hypertrophy in mice and observed the upregulation of KLK11 in hypertrophic hearts. Our in vitro experiments demonstrated that KLK11 overexpression promoted whereas KLK11 knockdown repressed cardiomyocytes hypertrophy induced by angiotensin II, as evidenced by cardiomyocyte size and the expression of hypertrophy-related fetal genes. Besides, we knocked down KLK11 expression in mouse hearts with adeno-associated virus 9. Knockdown of KLK11 in mouse hearts inhibited TAC-induced decline in fraction shortening and ejection fraction, reduced the increase in heart weight, cardiomyocyte size, and expression of hypertrophic fetal genes. We also observed that KLK11 promoted protein synthesis, the key feature of cardiomyocyte hypertrophy, by regulating the pivotal machines S6K1 and 4EBP1. Mechanism study demonstrated that KLK11 promoted the activation of AKT-mTOR signaling to promote S6K1 and 4EBP1 pathway and protein synthesis. Repression of mTOR with rapamycin blocked the effects of KLK11 on S6K1 and 4EBP1 as well as protein synthesis. Besides, rapamycin treatment blocked the roles of KLK11 in the regulation of cardiomyocyte hypertrophy. Conclusions Our findings demonstrated that KLK11 promoted cardiomyocyte hypertrophy by activating AKT-mTOR signaling to promote protein synthesis.

Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum)

2008 ◽  
Vol 375 (1) ◽  
pp. 150-152 ◽  
Author(s):  
Cheng Xin Yi ◽  
Jun Zhang ◽  
Ka Man Chan ◽  
Xiao Kun Liu ◽  
Yan Hong
Keyword(s):  
Gossypium Hirsutum ◽  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay ◽  
Transgene Copy Number

scholarly journals Virological Diagnosis of Herpes Simplex Virus 1 Esophagitis by Quantitative Real-Time PCR Assay

2011 ◽  
Vol 50 (3) ◽  
pp. 948-952 ◽  
Author(s):  
J.-F. Jazeron ◽  
C. Barbe ◽  
E. Frobert ◽  
F. Renois ◽  
D. Talmud ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay ◽  
Herpes Simplex Virus 1 ◽  
Virological Diagnosis ◽  
Simplex Virus

scholarly journals Regulation of virulence and β-lactamase gene expression in Staphylococcus aureus isolates: cooperation of two-component systems in bloodstream superbugs

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sanaz Dehbashi ◽  
Hamed Tahmasebi ◽  
Behrouz Zeyni ◽  
Mohammad Reza Arabestani
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Gene Expressions ◽  
Resistance Development ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Lactamase Gene

Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA)-bloodstream infections (BSI) are predominantly seen in the hospital or healthcare-associated host. Nevertheless, the interactions of virulence factor (VFs) regulators and β-lactam resistance in MRSA-BSI are unclear. This study aims to characterize the molecular relationship of two-component systems of VFs and the expression of the β-lactamase gene in MRSA-BSI isolates. In this study, 639 samples were collected from BSI and identified by phenotypic methods. We performed extensive molecular characterization, including SCCmec type, agr type, VFs gene profiles determinations, and MLST on isolates. Also, a quantitative real-time PCR (q-RT PCR) assay was developed for identifying the gene expressions. Results Ninety-one (91) S. aureus and 61 MRSA (67.0%) strains were detected in BSI samples. The presence of VFs and SCCmec genes in MRSA isolates were as follows: tst (31.4%), etA (18.0%), etB (8.19%), lukS-PVL (31.4%), lukF-PV (18.0%), lukE-lukD (16.3%), edin (3.2%), hla (16.3%), hlb (18.0%), hld (14.7%), hlg (22.9%), SCCmecI (16.3%), SCCmecII (22.9%), SCCmecIII (36.0%), SCCmecIV (21.3%), and SCCmecV (16.3%). Quantitative real-time PCR showed overexpression of mecRI and mecI in the toxigenic isolates. Moreover, RNAIII and sarA genes were the highest expressions of MRSA strains. The multi-locus sequence typing data confirmed a high prevalence of CC5, CC8, and CC30. However, ST30, ST22, and ST5 were the most prevalent in the resistant and toxigenic strains. Conclusion We demonstrated that although regulation of β-lactamase gene expressions is a significant contributor to resistance development, two-component systems also influence antibiotic resistance development in MRSA-BSI isolates. This indicates that resistant strains might have pathogenic potential. We also confirmed that some MLST types are more successful colonizers with a potential for MRSA-BSI.

Diagnosis of neuroschistosomiasis by antibody specificity index and semi-quantitative real-time PCR from cerebrospinal fluid and serum

2014 ◽  
Vol 63 (2) ◽  
pp. 309-312 ◽  
Author(s):  
Georg Härter ◽  
Hagen Frickmann ◽  
Sebastian Zenk ◽  
Dominic Wichmann ◽  
Bettina Ammann ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Symptoms ◽  
Diagnostic Procedures ◽  
Lower Limbs ◽  
Antibody Specificity ◽  
Quantitative Real Time Pcr ◽  
Specific Antibodies ◽  
Routine Diagnosis

We describe the case of a 16-year-old German male expatriate from Ghana who presented with obstipation, dysuria, dysaesthesia of the gluteal region and the lower limbs, bilateral plantar hypaesthesia and paraesthesia without pareses. A serum–cerebrospinal fluid (CSF) Schistosoma spp. specific antibody specificity index of 3.1 was considered highly suggestive of intrathecal synthesis of anti-Schistosoma spp. specific antibodies, although standardization of this procedure has not previously been described. Diagnosis was confirmed by detection of Schistosoma DNA in CSF by semi-quantitative real-time PCR at 100-fold concentration compared with serum. Accordingly the two diagnostic procedures, which have not previously been applied for routine diagnosis, appear to be useful for the diagnosis of neuroschistosomiasis. Clinical symptoms resolved following anthelmintic and anti-inflammatory therapy.

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