Prompt detection of endogenous hypochlorite (ClO−) in murine macrophages and zebrafish embryos facilitated by a distinctive chemodosimetric mode

2020 ◽  
Vol 18 (34) ◽  
pp. 6716-6723
Author(s):  
Shrabani Saha ◽  
Sujoy Das ◽  
Sriparna Das ◽  
Anwesha Samanta ◽  
Sudipta Maitra ◽  
...  
Keyword(s):  
Danio Rerio ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Zebrafish Embryos ◽  
Murine Macrophages ◽  
Naphthalene Diimide

A fluorescein appended naphthalene diimide based probe (FANDI) has been developed to selectively recognize exogenous as well as endogenous ClO− ions in RAW 264.7 cells (macrophages) and zebrafish embryos (Danio rerio).

scholarly journals Burkholderia pseudomallei Type III Secretion System Mutants Exhibit Delayed Vacuolar Escape Phenotypes in RAW 264.7 Murine Macrophages

2008 ◽  
Vol 76 (7) ◽  
pp. 2991-3000 ◽  
Author(s):  
Mary N. Burtnick ◽  
Paul J. Brett ◽  
Vinod Nair ◽  
Jonathan M. Warawa ◽  
Donald E. Woods ◽  
...  
Keyword(s):  
Type Iii Secretion ◽  
Burkholderia Pseudomallei ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Triple Mutant ◽  
Necrosis Factor Alpha ◽  
Murine Macrophages ◽  
Type Iii

ABSTRACT Burkholderia pseudomallei is a facultative intracellular pathogen capable of surviving and replicating within eukaryotic cells. Recent studies have shown that B. pseudomallei Bsa type III secretion system 3 (T3SS-3) mutants exhibit vacuolar escape and replication defects in J774.2 murine macrophages. In the present study, we characterized the interactions of a B. pseudomallei bsaZ mutant with RAW 264.7 murine macrophages. Following uptake, the mutant was found to survive and replicate within infected RAW 264.7 cells over an 18-h period. In addition, high levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF), and RANTES, but not IL-1α and IL-1β, were detected in culture supernatants harvested from infected monolayers. The subcellular location of B. pseudomallei within infected RAW 264.7 cells was determined, and as expected, the bsaZ mutant demonstrated early-vacuolar-escape defects. Interestingly, however, experiments also indicated that this mutant was capable of delayed vacuolar escape. Consistent with this finding, evidence of actin-based motility and multinucleated giant cell formation were observed between 12 and 18 h postinfection. Further studies demonstrated that a triple mutant defective in all three B. pseudomallei T3SSs exhibited the same phenotype as the bsaZ mutant, indicating that functional T3SS-1 and T3SS-2 did not appear to be responsible for the delayed escape phenotype in RAW 264.7 cells. Based upon these findings, it appears that B. pseudomallei may not require T3SS-1, -2, and -3 to facilitate survival, delayed vacuolar escape, and actin-based motility in activated RAW 264.7 macrophages.

scholarly journals Burkholderia mallei Cluster 1 Type VI Secretion Mutants Exhibit Growth and Actin Polymerization Defects in RAW 264.7 Murine Macrophages

2009 ◽  
Vol 78 (1) ◽  
pp. 88-99 ◽  
Author(s):  
Mary N. Burtnick ◽  
David DeShazer ◽  
Vinod Nair ◽  
Frank C. Gherardini ◽  
Paul J. Brett
Keyword(s):  
Fluorescence Microscopy ◽  
Actin Polymerization ◽  
Severe Disease ◽  
Critical Role ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Burkholderia Mallei ◽  
Type Vi Secretion System ◽  
Murine Macrophages ◽  
Type Vi Secretion

ABSTRACT Burkholderia mallei is a facultative intracellular pathogen that causes severe disease in animals and humans. Recent studies have shown that the cluster 1 type VI secretion system (T6SS-1) expressed by this organism is essential for survival in a hamster model of glanders. To better understand the role of T6SS-1 in the pathogenesis of disease, studies were initiated to examine the interactions of B. mallei tssE mutants with RAW 264.7 murine macrophages. Results obtained by utilizing modified gentamicin protection assays indicated that although the tssE mutants were able to survive within RAW 264.7 cells, significant growth defects were observed in comparison to controls. In addition, analysis of infected monolayers by differential interference contrast and fluorescence microscopy demonstrated that the tssE mutants lacked the ability to induce multinucleated giant cell formation. Via the use of fluorescence microscopy, tssE mutants were shown to undergo escape from lysosome-associated membrane protein 1-positive vacuoles. Curiously, however, following entry into the cytosol, the mutants exhibited actin polymerization defects resulting in inefficient intra- and intercellular spread characteristics. Importantly, all mutant phenotypes observed in this study could be restored by complementation. Based upon these findings, it appears that T6SS-1 plays a critical role in growth and actin-based motility following uptake of B. mallei by RAW 264.7 cells.

Valsartan, independently of AT1 receptor or PPARγ, suppresses LPS-induced macrophage activation and improves insulin resistance in cocultured adipocytes

2012 ◽  
Vol 302 (3) ◽  
pp. E286-E296 ◽  
Author(s):  
Misaki Iwashita ◽  
Hideyuki Sakoda ◽  
Akifumi Kushiyama ◽  
Midori Fujishiro ◽  
Haruya Ohno ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Angiotensin Ii ◽  
Insulin Signaling ◽  
Raw 264.7 Cells ◽  
Angiotensin Ii Receptor Blocker ◽  
Raw 264.7 ◽  
Peroxisome Proliferator ◽  
Angiotensin Ii Receptor ◽  
Murine Macrophages ◽  
At1a Receptor

Macrophages are integrated into adipose tissues and interact with adipocytes in obese subjects, thereby exacerbating adipose insulin resistance. This study aimed to elucidate the molecular mechanism underlying the insulin-sensitizing effect of the angiotensin II receptor blocker (ARB) valsartan, as demonstrated in clinical studies. Insulin signaling, i.e., insulin receptor substrate-1 and Akt phosphorylations, in 3T3-L1 adipocytes was impaired markedly by treatment with tumor necrosis factor-α (TNFα) or in the culture medium of lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages, and valsartan had no effects on these impairments. However, in contrast, when cocultured with RAW 264.7 cells using a transwell system, the LPS-induced insulin signaling impairment in 3T3-L1 adipocytes showed almost complete normalization with coaddition of valsartan. Furthermore, valsartan strongly suppressed LPS-induced productions of cytokines such as interleukin (IL)-1β, IL-6, and TNFα with nuclear factor-κB activation and c-Jun NH2-terminal kinase phosphorylation in RAW 264.7 and primary murine macrophages. Very interestingly, this effect of valsartan was also observed in THP-1 cells treated with angiotensin II type 1 (AT1) siRNA or a peroxisome proliferator-activated receptor-γ (PPARγ) antagonist as well as macrophages from AT1a receptor-knockout mice. We conclude that valsartan suppresses the inflammatory response of macrophages, albeit not via PPARγ or the AT1a receptor. This suppression appears to secondarily improve adipose insulin resistance.

scholarly journals Fabrication and characterization of Agarwood extract-loaded nanocapsules and evaluation of their toxicity and anti-inflammatory activity on RAW 264.7 cells and in zebrafish embryos

Drug Delivery ◽  
2021 ◽  
Vol 28 (1) ◽  
pp. 2618-2633
Author(s):  
Manar A. Eissa ◽  
Yumi Z. H.-Y. Hashim ◽  
Mohd Hamzah Mohd Nasir ◽  
Yusilawati Ahmad Nor ◽  
Hamzah Mohd. Salleh ◽  
...  
Keyword(s):  
Inflammatory Activity ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Zebrafish Embryos ◽  
Fabrication And Characterization ◽  
Anti Inflammatory

scholarly journals Cytotoxicity of Eurycomanone and Fargesin in RAW 264.7 Murine Macrophages

2021 ◽  
pp. 275-284
Author(s):  
Hanim Akmar Rosly ◽  
Amira Nabila Mat Roof ◽  
Salfarina Ramli ◽  
Visarut Buranasudja ◽  
Pornchai Rojsitthisak ◽  
...  
Keyword(s):  
Inhibitory Concentration ◽  
Morphological Changes ◽  
New Drugs ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Murine Macrophages ◽  
Eurycoma Longifolia ◽  
Anti Inflammatory ◽  
Inflammatory Effect

Bioactive natural compounds derived from plants are the source for the development of new drugs. Numerous in vitro studies have explored the anti-inflammatory effect of eurycomanone and fargesin, derived from Eurycoma longifolia and Flosmagnoliae, respectively. However, before anti-inflammatory investigation is conducted, it is important to obtain the safe doses of these compounds to ensure the validity of the anti-inflammatory results. Therefore, the present study was aimed to investigate the cytotoxicity of eurycomanone and fargesin towards macrophage RAW 264.7. cells to determine the safe doses of these compounds. Different concentrations of eurycomanone and fargesin were subjected to RAW 264.7 cells. The cytotoxicity of the compounds was evaluated by MTT assay and 50% inhibitory concentration (IC50) of these compounds was determined. Morphological changes of RAW 264.7 cells upon exposure to these compounds were also observed. Eurycomanone exhibited its cytotoxic effect by reducing RAW 264.7 cell viability dose-dependently with the IC50 of 94.17 µM. Meanwhile, fargesin had slight cytotoxicity towards RAW 264.7 cells with the IC50 of 173.5µM. Eurycomanone was more cytotoxic towards RAW 264.7 cells compared to fargesin. In conclusion, eurycomanone and fargesin at concentration up to 25 µM, was not toxic to the RAW 264.7 murine macrophages cells and the findings can be applied in the future anti-inflammatory study.

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