A simple method for assaying extracellular hydroxyl radical activity and its application to natural and synthetic waters

2003 ◽  
Vol 60 (2) ◽  
pp. 203-213 ◽  
Author(s):  
L A Molot ◽  
S A Miller ◽  
P J Dillon ◽  
C G Trick

An assay has been developed to measure extracellular hydroxyl radical (OH*) activity in algal culture media and natural waters over a 4- to 5-day period. The first-order rate constant, k, for loss of absorbance at 590 or 620 nm was determined for erioglaucine, which is sensitive to OH*, insensitive to superoxide and hydrogen peroxide, and stable in the dark and under artificial radiation (280–750 nm) and solar radiation in the absence of oxidants. Variation in irradiance was accounted for by normalizing k with k for a ferric iron reference solution with dye (k/kfe). Trends in k/kfe for streams and lakes were consistent with previous data on photochemical oxidation rates of dissolved organic matter. Values for k/kfe were similar in filtered surface waters of eutrophic Heart Lake and nearby mesotrophic Lake St. George under artificial radiation. Hence, extracellular OH* did not appear to be a direct cause of the onset of a nuisance cyanobacterial bloom in Heart Lake, nor did OH* appear related to the absence of a bloom in Lake St. George. k/kfe was two orders of magnitude higher in algal culture media supplied with 8.8 mM nitrate than in lake waters.

2018 ◽  
Author(s):  
Joshua S. Sharp ◽  
Sandeep K. Misra ◽  
Jeffrey J. Persoff ◽  
Robert W. Egan ◽  
Scot R. Weinberger

AbstractHydroxyl radical protein footprinting (HRPF) is a powerful method for measuring protein topography, allowing researchers to monitor events that alter the solvent accessible surface of a protein (e.g. ligand binding, aggregation, conformational changes, etc.) by measuring changes in the apparent rate of reaction of portions of the protein to hydroxyl radicals diffusing in solution. Fast Photochemical Oxidation of Proteins (FPOP) offers an ultra-fast benchtop method for performing HRPF, photolyzing hydrogen peroxide using a UV laser to generate high concentrations of hydroxyl radicals that are consumed on roughly a microsecond timescale. The broad reactivity of hydroxyl radicals means that almost anything added to the solution (e.g. ligands, buffers, excipients, etc.) will scavenge hydroxyl radicals, altering their half-life and changing the effective radical concentration experienced by the protein. Similarly, minute changes in peroxide concentration, laser fluence, and buffer composition can alter the effective radical concentration, making reproduction of data challenging. Here, we present a simple method for radical dosimetry that can be carried out as part of the FPOP workflow, allowing for measurement of effective radical concentration in real time. Additionally, by modulating the amount of radical generated, we demonstrate that FPOP HRPF experiments carried out in buffers with widely differing levels of hydroxyl radical scavenging capacity can be normalized on the fly, yielding statistically indistinguishable results for the same conformer. This method represents a major step in transforming FPOP into a robust and reproducible technology capable of probing protein structure in a wide variety of contexts.


2021 ◽  
Vol 105 (5) ◽  
pp. 2139-2156
Author(s):  
Justine Sauvage ◽  
Gary H. Wikfors ◽  
Xiaoxu Li ◽  
Mark Gluis ◽  
Nancy Nevejan ◽  
...  

Abstract The efficiency of microalgal biomass production is a determining factor for the economic competitiveness of microalgae-based industries. N-acetylcysteine (NAC) and pluronic block polymers are two compounds of interest as novel culture media constituents because of their respective protective properties against oxidative stress and shear-stress-induced cell damage. Here we quantify the effect of NAC and two pluronic (F127 and F68) culture media additives upon the culture productivity of six marine microalgal species of relevance to the aquaculture industry (four diatoms-Chaetoceros calcitrans, Chaetoceros muelleri, Skeletonema costatum, and Thalassiosira pseudonana; two haptophytes-Tisochrysis lutea and Pavlova salina). Algal culture performance in response to the addition of NAC and pluronic, singly or combined, is dosage- and species-dependent. Combined NAC and pluronic F127 algal culture media additives resulted in specific growth rate increases of 38%, 16%, and 24% for C. calcitrans, C. muelleri, and P. salina, respectively. Enhanced culture productivity for strains belonging to the genus Chaetoceros was paired with an ~27% increase in stationary-phase cell density. For some of the species examined, culture media enrichments with NAC and pluronic resulted in increased omega-3-fatty acid content of the algal biomass. Larval development (i.e., growth and survival) of the Pacific oyster (Crassostrea gigas) was not changed when fed a mixture of microalgae grown in NAC- and F127-supplemented culture medium. Based upon these results, we propose that culture media enrichment with NAC and pluronic F127 is an effective and easily adopted approach to increase algal productivity and enhance the nutritional quality of marine microalgal strains commonly cultured for live-feed applications in aquaculture. Key points • Single and combined NAC and pluronic F127 culture media supplementation significantly enhanced the productivity of Chaetoceros calcitrans and Chaetoceros muelleri cultures. • Culture media enrichments with NAC and F127 can increase omega-3-fatty acid content of algal biomass. • Microalgae grown in NAC- and pluronic F127-supplemented culture media are suitable for live-feed applications.


2021 ◽  
Author(s):  
Christine Poon

AbstractArthroplasty implants e.g. hip, knee, spinal disc sustain relatively high compressive loading and friction wear, which lead to the formation of wear particles or debris between articulating surfaces. Despite advances in orthopaedic materials and surface treatments, the production of wear debris from any part of a joint arthroplasty implant is currently unavoidable. Implant wear debris induces host immune responses and inflammation, which causes patient pain and ultimately implant failure through progressive inflammation-mediated osteolysis and implant loosening, where the severity and rate of periprosthetic osteolysis depends on the material and physicochemical characteristics of the wear particles. Evaluating the cytotoxicity of implant wear particles is important for regulatory approved clinical application of arthroplasty implants, as is the study of cell-particle response pathways. However, the wear particles of polymeric materials commonly used for arthroplasty implants tend to float when placed in culture media, which limits their contact with cell cultures. This study reports a simple means of suspending wear particles in liquid medium using sodium carboxymethyl cellulose (NaCMC) to provide a more realistic proxy of the interaction between cells and tissues to wear particles in vivo, which are free-floating in synovial fluid within the joint cavity. Low concentrations of NaCMC dissolved in culture medium were found to be effective for suspending polymeric wear particles. Such suspensions may be used as more physiologically-relevant means for testing cellular responses to implant wear debris, as well as studying the combinative effects of shear and wear particle abrasion on cells in a dynamic culture environments such as perfused tissue-on-chip devices.


2016 ◽  
Vol 13 (4) ◽  
pp. 757 ◽  
Author(s):  
Jarod N. Grossman ◽  
Tara F. Kahan

Environmental contextReactions in natural waters such as lakes and streams are thought to be extremely slow in the absence of sunlight (e.g. at night). We demonstrate that in the presence of iron, hydrogen peroxide and certain bacteria (all of which are common in natural waters), certain reactions may occur surprisingly quickly. These findings will help us predict the fate of many compounds, including pollutants, in natural waters at night. AbstractDark Fenton chemistry is an important source of hydroxyl radicals (OH•) in natural waters in the absence of sunlight. Hydroxyl radical production by this process is very slow in many bodies of water, owing to slow reduction and low solubility of FeIII at neutral and near-neutral pH. We have investigated the effects of the iron-reducing bacteria Shewanella oneidensis (SO) on OH• production rates from Fenton chemistry at environmentally relevant hydrogen peroxide (H2O2) and iron concentrations at neutral pH. In the presence of 2.0 × 10–4M H2O2, OH• production rates increased from 1.3 × 10–10 to 2.0 × 10–10Ms–1 in the presence of 7.0 × 106cellsmL–1 SO when iron (at a concentration of 100μM) was in the form of FeII, and from 3.6 × 10–11 to 2.2 × 10–10Ms–1 when iron was in the form of FeIII. This represents rate increases of factors of 1.5 and 6 respectively. We measured OH• production rates at a range of H2O2 concentrations and SO cell densities. Production rates depended linearly on both variables. We also demonstrate that bacteria-assisted Fenton chemistry can result in rapid degradation of aromatic pollutants such as anthracene. Our results suggest that iron-reducing bacteria such as SO may be important contributors to radical formation in dark natural waters.


Minerals ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 74
Author(s):  
Chen Zhao ◽  
Pu Zhang ◽  
Xiangzhong Li ◽  
Youfeng Ning ◽  
Liangcheng Tan ◽  
...  

Four saline lakes in the northeast of Qaidam Basin were selected to explore the distribution characteristics and influencing factors of uranium isotopes in lake waters with high evaporation background. The 238U concentration and the activity ratios of 234U/238U ([234U/238U]AR) showed that there was no significant change in the same lake, but there was a certain degree of difference in the distribution between different lakes. We found that aqueous 238U concentration within a certain range increased with an increase in TDS (total dissolved solid) and salinity, as was also the case with pH. As in natural waters, the pH affects the speciation of 238U, but TDS and salinity affect the adsorption process of aqueous 238U. Further, the replenishment of water will also affect the uranium isotope concentration for lakes, but it is not the main influencing factor for saline lakes. Therefore, we suggest that pH is the dominant factor affecting changes in aqueous 238U concentration of the sampled saline lakes. The [234U/238U]AR in these saline lakes are closely related to the input water and the associated water–rock interactions involving sediments, atmosphere dust, and organic material, etc. during the evolution stage, metamorphous degree, and hydrochemistry of the saline lakes. Lake water samples collected in the maximum and minimum discharge water period, were used to evaluate the seasonal distribution characteristics of aqueous 238U, and we found that 238U concentration did not show an evident change with the seasons in these saline lakes. If the 238U concentration and [234U/238U]AR can remain consistent during a period of time, then the sediment ages and/or sedimentation rates could be determined by lake sediment and/or biogenic carbonate in future, thus allowing for the accurate reconstruction of the paleoclimate and paleoenvironment.


2020 ◽  
Vol 54 (21) ◽  
pp. 14096-14106
Author(s):  
Xiaohui Zhu ◽  
William L. Miller ◽  
Cédric G. Fichot

Oxidation of atmospheric sulphur dioxide can occur by homogeneous photochemically initiated gas-phase reactions as well as by heterogeneous reactions in cloud and fog droplets. Gas phase oxidation can result from reactions of excited SO 2 molecules formed by absorption of solar u.v. radiation by ground state SO 2 , from reactions of SO 2 with photochemically generated OH and RO 2 free radicals, and from its reaction with transient species produced in thermal ozone—alkene reactions. Evaluation of the available mechanistic and rate data reveals that, of these three processes, oxidation by free radicals, particularly OH, is likely to be the most important in the atmosphere. Oxidation rates of up to 4 % h -1 are predicted for a hydrocarbon-NO x polluted atmosphere under western European summertime conditions. This can lead to the formation of elevated concentrations of sulphuric acid and sulphate aerosol in polluted air. In the natural background troposphere oxidation rates are much less, ca . 0.3 % h -1 averaged over 24 h, but probably still significant as a source of atmospheric sulphates.


2014 ◽  
Vol 16 (4) ◽  
pp. 757-763 ◽  
Author(s):  
Luni Sun ◽  
Hongmei Chen ◽  
Hussain A. Abdulla ◽  
Kenneth Mopper

This study showed that hydroxyl radical (˙OH) production during long-term irradiation experiments is most accurately measured using instantaneous rates.


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