Screening of Actinobacterial Extracts/Compounds for Antimycobacterial Activity by Luciferase Reporter Phage (LRP) Assay

2022 ◽  
pp. 395-399
Author(s):  
Shuai Wei ◽  
Shucheng Liu ◽  
Ramachandran Chelliah ◽  
Deog-Hwan Oh
Keyword(s):  
Antimycobacterial Activity ◽  
Luciferase Reporter

SYNTHESIS AND ANTIMYCOBACTERIAL ACTIVITY OF BENZIMIDAZOLE DERIVATIVES

INDIAN DRUGS ◽  
2012 ◽  
Vol 49 (02) ◽  
pp. 17-25
Author(s):  
P. S Mathapati ◽  
◽  
S. A. Durgad ◽  
K. C. Singh ◽  
S. D. Bhinge
Keyword(s):  
Aromatic Aldehydes ◽  
Antimycobacterial Activity ◽  
Luciferase Reporter ◽  
Benzimidazole Derivatives ◽  
Dilution Method ◽  
Preliminary Screening ◽  
Spectral Techniques ◽  
Antibacterial And Antifungal ◽  
Diffusion Assay

In the present study, the objective is to synthesize novel compounds from the benzimidazole moiety,recognized for its various biological potentials. Chalcone being a very good synthon was utilized to combine with the benzimidazole moiety to design agents with significant antimycobacterial activity. Benzimidazole is formed when o-phenylene diamine is heated with organic acid i.e. formic acid; in presence of strong alkali sodium hydroxide it will gives benzimidazole. Benzimidazole was acetylated with acetyl chloride, the intermediate substituted chalcones were synthesized from acetylated benzimidazole, in order to synthesize different substituted aromatic aldehydes of the basic moiety, 1-sub. benzimidazole to obtain the different benzimidazole derivatives. Their structures were characterized by various spectral techniques like IR, NMR and MS. For the preliminary screening (antibacterial and antifungal)for antimicrobial activity, the test was performed by disc diffusion assay and determination of minimum inhibitory concentration (MIC) by broth dilution method. The antimycobacterial screening was performed against Mycobacterium tuberculosis by Almar blue assay (ABA) and Luciferase reporter phase (LRPA) techniques using isoniazid as the standard.

scholarly journals Antimycobacterial activity of certain mangrove plants against multi-drug resistant Mycobacterium tuberculosis

2014 ◽  
Vol 5 (3) ◽  
pp. 54-57 ◽  
Author(s):  
Amudha Prabu ◽  
Prabu Seenivasan ◽  
Vanaja Kumar
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Tamil Nadu ◽  
Medical Science ◽  
Antimycobacterial Activity ◽  
Luciferase Reporter ◽  
Phytochemical Analysis ◽  
Sesuvium Portulacastrum ◽  
Mangrove Plant ◽  
Mangrove Plants ◽  
Study Results

Objectives: The present study was conducted aiming to identify a potent mangrove plant against Mycobacterium tuberculosis using rapid method, luciferase reporter phage (LRP) assay. Methods: Seven mangrove plants viz., Ceriops decandra, Aegiceras corniculatum, Excoecaria agollacha,Avicennia officinalis, Rhizophora mucronata, Suaeda monoica and Sesuvium portulacastrum were collected from Pichavaram mangrove forest in the east coast of, Tamil Nadu. Hexane and methanol extracts were obtained and antimycobacterial activity was done by LRP assay at the concentration of 500 μg/ml. Mycobacterial strains viz. reference strain M. tuberculosis H37Rv and two clinical isolates of M. tuberculosis, one sensitive and the other resistant to Streptomycin, Isoniazid, Rifampicin and Ethambutol used for this study. Results: Methanol extract of E. agollacha showed maximum antimycobacterial activity followed by A. corniculatum and A. officinalis at 500 μg/ml concentration. The remaining four plants (Suaeda monoica, Sesuvium portulacastrum, R. mucronata and C. decandra) showed less or no activity. However, hexane extracts of all the tested plants failed to inhibit the mycobacterial strains. Conclusions: Among the seven plants tested, E. agollacha was found to have better anti-TB activity. Further studies based on bioassay guided fractionation and phytochemical analysis may useful to identify the potent antimycobacterial molecule(s) from this mangrove plant. Asian Journal of Medical Science, Volume-5(3) 2014: 54-57 http://dx.doi.org/10.3126/ajms.v5i3.9171 

scholarly journals LUCIFERASE REPORTER MYCOBACTERIOPHAGES FOR EVALUATING NORBORNENE-BASED ANTITUBERCULOSIS DRUG SUSCEPTIBILITY TESTING ON MYCOBACTERIUM TUBERCULOSIS

2017 ◽  
Vol 10 (9) ◽  
pp. 406
Author(s):  
Kalaiselvi K ◽  
Mangayarkarasi V ◽  
Gomathi Ns ◽  
Balaji S ◽  
Shivshankar R Mane ◽  
...  
Keyword(s):  
Resistant Strain ◽  
Antimycobacterial Activity ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Luciferase Reporter ◽  
Concentration Method ◽  
Poly Ethylene ◽  
Low Dosage

  Objective: In 2015, 9.6 million people around the world became sick with tuberculosis (TB) disease and 1.5 million TB-related deaths worldwide. Recent increasing incidence of multidrug-resistant (MDR; resistance to at least rifampicin (RIF) and isoniazid [INH]) and extensively drug-resistant (MDR resistance plus resistance to a fluoroquinolone and an aminoglycoside) makes TB a serious concern. Lot of research is needed to deal with this infectious disease for a better alternative in treatment or modification of these older TB drugs. The present study aimed at evaluating antimycobacterial activity of norbornene (NOR) derived INH copolymer with poly ethylene glycol (NOR- polyethylene glycol [PEG]-INH) a novel nanocarrier along with the anti-TB drug using luciferase reporter phages (LRPs).Methods: NOR derived INH accounts for 74% of INH, 24% of NOR, and 2% of PEG. H37Rv control strain, a sensitive, and a resistant strain of Mycobacterium TB (MTB) used in this study. The in vitro activity of the drug was evaluated using absolute concentration method. The resistant strain was evaluated using LRP assay to observe the minimum inhibitory concentration of the drug.Results: Reduction in light units was observed for the resistant strain exposed to plain INH and NOR-PEG-INH separately. 24% of reduction was observed in strains exposed to plain INH whereas 37% of reduction was observed in strains exposed to NOR-PEG-INH.Conclusion: NOR-based INH had better antimycobacterial activity compared to plain INH and RIF. Antimycobacterial activity of INH and RIF increases even with very low dosage with NOR conjugate.

scholarly journals 1828. Bedaquiline Resistance in Mycobacterium intracellulare Is Mediated by the Transcriptional Repressor MmpT5

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S41-S41
Author(s):  
David Alexander ◽  
Joshua Yoneda ◽  
Supriya Bhat ◽  
Teagan Oleksyn ◽  
Jeffrey Chen ◽  
...  
Keyword(s):  
Antimycobacterial Activity ◽  
Luciferase Reporter ◽  
Drug Efflux ◽  
Bacterial Survival ◽  
Wild Type ◽  
Luciferase Reporter Gene ◽  
Low Level ◽  
Pre Treatment ◽  
High Level ◽  
Level Resistance

Abstract Background Bedaquiline (BDQ) is an FDA approved antibiotic with antimycobacterial activity. BDQ resistance has been observed in several Mycobacterium species. High-level resistance is due to mutations in ATP synthase. Low -level resistance is attributed to drug efflux. Previously, we suggested that the MmpSL5 efflux system mediates BDQ resistance in M. intracellulare. Here, we examine the role of MmpT5 in transcriptional regulation of mmpSL5 and BDQ resistance. Methods In this study, mmpSL5-mmpT5 genes were cloned from 2 pre-treatment (wild-type mmpT5) and 2 relapse (mutant mmpT5) isolates of M. intracellulare and transformed into M. smegmatis. BDQ MICs were determined as well as cell survival after 24 hours exposure to an inhibitory concentration (0.07 µg/mL) of BDQ. Transcription of the M. intracellulare mmpT5 and mmpSL5 promoters was monitored with luciferase reporter gene fusions in the presence of wild-type and mutant alleles of mmpT5. Single and multigene constructs were created using the MoClo system, and transformed into E. coli DH5α. Constructs containing the M. tuberculosis rv0678 gene, which mediates low-level BDQ resistance in M. tuberculosis, were also examined. Results The BDQ MIC for the M. smegmatis control strain, and all strains containing mmpSL5-mmpT5 constructs, was 0.007 µg/mL. Even so, strains containing mutant mmpT5 alleles showed enhanced survival after 24 hours exposure to 0.007 µg/mL BDQ. Bacterial colonies associated with mutant mmpT5 alleles exhibited altered morphology relative to wild-type strains. Transcription of mmpSL5 was repressed by wild-type mmpT5, but neither mutant mmpT5 nor rv0678 repressed transcription. The mmpT5 luciferase reporter was not active. Conclusion MmpT5 represses transcription of mmpSL5 whereas the operon is dysregulated by mmpT5 mutations. Although Rv0678 regulates mmpSL expression in M. tuberculosis, it cannot repress the M. intracellulare mmpSL5 genes. The mmpSL5-mmpT5 genes have no impact on the BDQ MIC for M. smegmatis, but constructs containing mutant mmpT5 alleles do enhance bacterial survival. The altered morphology of these colonies suggests that BDQ resistance is mediated by cell wall changes in combination with drug efflux. Disclosures All Authors: No reported Disclosures.

The Anti-Atherogenic Properties of Sesamin are Mediated via Improved Macrophage Cholesterol Efflux Through PPARγ1-LXRα and MAPK Signaling

2014 ◽  
Vol 84 (1-2) ◽  
pp. 79-91 ◽  
Author(s):  
Amin F. Majdalawieh ◽  
Hyo-Sung Ro
Keyword(s):  
Cholesterol Efflux ◽  
Transcriptional Activity ◽  
Molecular Mechanisms ◽  
Foam Cell ◽  
Mapk Signaling ◽  
Luciferase Reporter ◽  
Foam Cell Formation ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Macrophage Cholesterol Efflux

Background: Foam cell formation resulting from disrupted macrophage cholesterol efflux, which is triggered by PPARγ1 and LXRα, is a hallmark of atherosclerosis. Sesamin and sesame oil exert anti-atherogenic effects in vivo. However, the exact molecular mechanisms underlying such effects are not fully understood. Aim: This study examines the potential effects of sesamin (0, 25, 50, 75, 100 μM) on PPARγ1 and LXRα expression and transcriptional activity as well as macrophage cholesterol efflux. Methods: PPARγ1 and LXRα expression and transcriptional activity are assessed by luciferase reporter assays. Macrophage cholesterol efflux is evaluated by ApoAI-specific cholesterol efflux assays. Results: The 50 μM, 75 μM, and 100 μM concentrations of sesamin up-regulated the expression of PPARγ1 (p< 0.001, p < 0.001, p < 0.001, respectively) and LXRα (p = 0.002, p < 0.001, p < 0.001, respectively) in a concentration-dependent manner. Moreover, 75 μM and 100 μM concentrations of sesamin led to 5.2-fold (p < 0.001) and 6.0-fold (p<0.001) increases in PPAR transcriptional activity and 3.9-fold (p< 0.001) and 4.2-fold (p < 0.001) increases in LXR transcriptional activity, respectively, in a concentration- and time-dependent manner via MAPK signaling. Consistently, 50 μM, 75 μM, and 100 μM concentrations of sesamin improved macrophage cholesterol efflux by 2.7-fold (p < 0.001), 4.2-fold (p < 0.001), and 4.2-fold (p < 0.001), respectively, via MAPK signaling. Conclusion: Our findings shed light on the molecular mechanism(s) underlying sesamin’s anti-atherogenic effects, which seem to be due, at least in part, to its ability to up-regulate PPARγ1 and LXRα expression and transcriptional activity, improving macrophage cholesterol efflux. We anticipate that sesamin may be used as a therapeutic agent for treating atherosclerosis.

Influence of chlorophyll and tannins in plant extracts on cell-based luciferase reporter gene assays

Planta Medica ◽  
2009 ◽  
Vol 75 (09) ◽  
Author(s):  
S Vogl ◽  
P Picker ◽  
N Fakhrudin ◽  
A Atanasov ◽  
E Heiß ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Plant Extracts ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Reporter Gene Assays

Antimycobacterial activity of medicinal plants from Mozambique

Planta Medica ◽  
2009 ◽  
Vol 75 (09) ◽  
Author(s):  
G Da Silva ◽  
A Macedo ◽  
I Famba ◽  
M Taniça ◽  
R Serrano ◽  
...  
Keyword(s):  
Medicinal Plants ◽  
Antimycobacterial Activity

Post-Transcriptional Regulation of Cardiac Sarcomere Protein Titin Through 3’ Untranslated Regions

2013 ◽  
Vol 9 ◽  
Author(s):  
Tara A Shrout
Keyword(s):  
Gene Expression ◽  
Transcriptional Control ◽  
Striated Muscle ◽  
Binding Capacity ◽  
Structural Features ◽  
Neonatal Rat ◽  
Regulatory Control ◽  
Untranslated Regions ◽  
Luciferase Reporter ◽  
Regulatory Effects

Titin is the largest known protein in the human body, and forms the backbone of all striated muscle sarcomeres. The elastic nature of titin is an important component of muscle compliance and functionality. A significant amount of energy is expended to synthesize titin, thus we postulate that titin gene expression is under strict regulatory control in order to conserve cellular resources. In general, gene expression is mediated in part by post-transcriptional control elements located within the 5’ and 3’ untranslated regions (UTRs) of mature mRNA. The 3’UTR in particular contains structural features that affect binding capacity to other RNA components, such as MicroRNA, which control mRNA localization, translation, and degradation. The degree and significance of the regulatory effects mediated by two determined variants of titin’s 3’ UTR were evaluated in Neonatal Rat Ventricular Myocyte and Human Embryonic Kidney cell lines. Recombinant plasmids to transfect these cells lines were engineered by insertion of the variant titin 3’UTR 431- and 1047-base pairs sequences into luciferase reporter vectors. Expression due to an unaltered reporter vector served as the control. Quantitative changes in luciferase activity due to the recombinants proportionally represented the effect titin’s respective 3’UTR conferred on downstream post-transcriptional expression relative to the control. The effect due to titin’s shorter 3’UTR sequence was inconclusive; however, results illustrated that titin’s longer 3’UTR sequence caused a 35 percent decrease in protein expression. Secondary structural analysis of the two sequences revealed differential folding patterns that affect the stability and degree of MicroRNA-binding within titin’s variant 3’UTR sequences.

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