Measurement of nitrate and nitrite in biological samples using nitrate reductase and Griess reaction

Granger, Donald L.; Taintor, Read R.; Boockvar, Kenneth S.; Hibbs, John B.

doi:10.1016/s0076-6879(96)68016-1

Cited by 459 publications

(294 citation statements)

References 23 publications

Supporting

Mentioning

268

Contrasting

Unclassified

Order By: Relevance

“…Macrophage release of NO was measured in vitro by colorimetric assessment of NO 2 À , a stable metabolite (Granger et al 1996). In brief, after cell stimulation for 24 h with 1 lg LPS/ml alone or in the presence of increasing concentrations of pesticides, 50 ll culture supernatant was transferred from triplicate wells and placed into 96-well plates.…”

Section: Macrophage Nitrite Assaymentioning

confidence: 99%

Modulation of macrophage functionality induced in vitro by chlorpyrifos and carbendazim pesticides

Helali

Ferchichi

Maaouia

et al. 2016

Journal of Immunotoxicology

View full text Add to dashboard Cite

The immune response is the first defense against pathogens; however, it is very sensitive and can be impacted on by agrochemicals such as carbamate and organophosphate pesticides widely present in the environment. To understand how pesticides can affect immune cell function in vitro, this study investigated the effects of chlorpyrifos (CPF) and carbendazim (CBZ), the most commonly used pesticides worldwide, on murine immune cell (i.e. macrophage) functions, including lysosomal enzyme activity and proinflammatory cytokines (IL-1b and TNFa) and nitric oxide (NO) production by isolated mouse peritoneal macrophages. This study showed for the first time that CPF and CBZ dose-relatedly reduced macrophage lysosomal enzyme activity and LPS-induced production of IL-1b, TNFa and NO. In general, the effects caused by CPF appeared more pronounced than those by CBZ. Collectively, these results demonstrated that CPF and CBZ exhibited marked immunomodulatory effects and could act as potent immunosuppressive factors in vitro. This inhibition of macrophage pro-inflammatory function may be an integral part of the underlying mode of action related to pesticide-induced immunosuppression.ARTICLE HISTORY

show abstract

Section: Macrophage Nitrite Assaymentioning

confidence: 99%

Modulation of macrophage functionality induced in vitro by chlorpyrifos and carbendazim pesticides

Helali

Ferchichi

Maaouia

et al. 2016

Journal of Immunotoxicology

View full text Add to dashboard Cite

show abstract

“…To examine the amount of NO produced in the culture, we measured the concentration of nitrite, a stable NO metabolite, using Griess reagent as previously described (Granger et al, 1996). Briefly, 50 μl of the medium supernatant was mixed with 100 μl of Griess reagent, and the absorbance was measured at 550 nm using Fusion™ universal microplate analyser.…”

Section: Measurement Of Nitrite Concentrationmentioning

confidence: 99%

Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro

Kawakita

et al. 2006

Experimental Eye Research

View full text Add to dashboard Cite

Amniotic membrane (AM) used as a temporary or permanent graft for ocular surface reconstruction has a potent anti-inflammatory effect. We would like to investigate the mechanism whereby AM induces macrophage apoptosis in vitro. Mouse macrophages, Raw 264.7 cells, were cultured on plastic, type I collagen, corneal stromal slice or AM stromal matrix in serum-free medium with or without interferon-γ (IFN-γ). Cells were stained by LIVE/DEAD assay, Hoechst-33342, and TUNEL assay for cell death and apoptosis. Cell lysates and conditioned media were analysed by Cell Death Detection ELISA assay for quantitation of apoptosis. Conditioned media were also analysed by Griess assay for the nitrite concentration and ELISA assay for tumour necrosis factor alpha (IFN-α) concentration. Lysates of cells were subjected to Western blot analyses of IKK-α, IKK-β, p65 (RelA) subunit of nuclear factor κB (NF-κB), total Akt, phospho-Akt (Ser473), and phospho-FKHR (Thr24)/phosphor-FKHRL1 (Thr32). At 48 hr after cultivation, cells showed a low level of apoptosis when cultured on plastic, type I collagen and corneal stromal slice with or without IFN-γ and on AM without IFN-γ. Nevertheless, cells showed a significant increase of apoptosis when cultured on AM with IFN-γ activation, and this phenomenon became apparent only after 48 hr. IFN-γ-activated macrophages on plastic continuously produced nitric oxide (NO) and IFN-α during 72 hr culturing. In contrast, there was no NO and IFN-α production after 48 hr culture on AM. NO inhibitors, L-NMMA and L-NIL, attenuated NO production of IFN-γ-activated macrophages on AM, while apoptosis was not decreased accordingly. Expression of IKK-α, IKK-β, p65 (RelA) subunit of NF-κB total Akt, phosopho-Akt (Ser473), and phospho-FKHR (Thr24)/FKHRL1 (Thr32) was all down-regulated in IFN-γ-activated macrophages cultured on AM. In conclusion, AM stromal matrix induces apoptosis of IFN-γ activated, but not non-activated macrophages, not through the generation of NO, but instead by down-regulating anti-apoptotic NF-κB and Akt-FKHR signalling pathways.

show abstract

“…The production of nitrite by peritoneal macrophages was measured in culture supernatants at 24 h by the Griess assay (22). One hundred microliters of culture supernatants was mixed with 100 l of modified Griess reagent (Sigma) and incubated for 10 min.…”

Section: Nitrite Productionmentioning

confidence: 99%

TNF-α Controls Intracellular Mycobacterial Growth by Both Inducible Nitric Oxide Synthase-Dependent and Inducible Nitric Oxide Synthase-Independent Pathways

Bekker

Freeman

Murray

et al. 2001

The Journal of Immunology

140

View full text Add to dashboard Cite

The role of TNF-α in the control of mycobacterial growth in murine macrophages was studied in vitro. Infection of macrophages from TNF-α gene disrupted (TNF-knockout (KO)) mice with recombinant Mycobacterium bovis bacillus Calmette Guérin (BCG) expressing the vector only (BCG-vector) resulted in logarithmic growth of the intracellular bacilli. Infection with BCG-secreting murine TNF-α (BCG-TNF) led to bacillary killing. Killing of BCG-TNF was associated with rapid accumulation of inducible NO synthase (iNOS) protein and the production of nitrite. The uncontrolled growth of BCG-vector was associated with low iNOS expression but no nitrite production. Thus, iNOS expression appears to be TNF-α independent but iNOS generation of NO requires TNF-α. In cultures of TNF-KO macrophages infected with BCG-TNF, inhibition of iNOS by aminoguanidine (AMG) abolished the killing of the bacilli. However, the growth of the organisms was still inhibited, suggesting an iNOS-independent TNF-α-mediated growth inhibition. To confirm this, macrophages from iNOS-KO mice were infected with either BCG-vector or BCG-TNF. As expected, no nitrite was detected in the culture medium. TNF-α was detected only when the cells were infected with BCG-TNF. In the iNOS-KO macrophages, the growth of BCG was inhibited only in the BCG-TNF infection. These results suggest that in the absence of iNOS activity, TNF-α stimulates macrophages to control the growth of intracellular BCG. Thus, there appears to be both a TNF-α-dependent-iNOS-dependent killing pathway as well as a TNF-α-dependent-iNOS-independent growth inhibitory pathway for the control of intracellular mycobacteria in murine macrophages.

show abstract

Measurement of nitrate and nitrite in biological samples using nitrate reductase and Griess reaction

Cited by 459 publications

References 23 publications

Modulation of macrophage functionality induced in vitro by chlorpyrifos and carbendazim pesticides

Modulation of macrophage functionality induced in vitro by chlorpyrifos and carbendazim pesticides

Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro

TNF-α Controls Intracellular Mycobacterial Growth by Both Inducible Nitric Oxide Synthase-Dependent and Inducible Nitric Oxide Synthase-Independent Pathways

Contact Info

Product

Resources

About