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Access Microbiology 2019Honey is a natural product with many beneficial properties including antimicrobial action. Production of hydrogen peroxide (HO) in diluted honey is central to this...
Honey is a natural product with many beneficial properties including antimicrobial action. Production of hydrogen peroxide (HO) in diluted honey is central to this action. Here, we describe an optimized method for measuring levels of HO in honey. This method is based on established methods, with the level of dilution, the time between dilution and reading the assay, and aeration of the samples during the assay identified as critical points for ensuring reliability and reproducibility. The method is cost-effective and easy to perform using common laboratory equipment. Using this method, we quantified the hydrogen peroxide content of five different, unprocessed polyfloral honeys collected in NC, USA. Our results show that HO production by these honeys varies greatly, with some samples producing negligible levels of HO. We assessed the effect of colour on the assay by measuring the recovery of spiked HO from light and dark honey and from serially diluted dark corn syrup, and found the amount of HO that could be detected was lower in dark corn syrup and darker honey samples.
PubMed: 32974499
DOI: 10.1099/acmi.0.000065 -
Frontiers in Cell and Developmental... 2022Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development...
Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development and circulatory failure, suggesting additional roles for TF in embryonic development beyond coagulation. In this study, we characterized the role of one of the TF paralogs () using a zebrafish model. The expression of during embryonic developmental stages was determined by RT-PCR. Spatiotemporal expression pattern of revealed (high expression from 28 to 36 hpf) the role of in the development of the yolk sac, circulation, and fins. Morpholinos (MO), an antisense-based oligonucleotide strategy, was used to knockdown and examined for defects in morphological appearance, bleeding, and vascular patterning. MO-injected embryos showed morphological abnormalities, including shorter body lengths and crooked tails. O-dianisidine staining showed MO-injected embryos exhibited bleeding in the trunk (5.44%) and head (9.52%) regions. Imaging of endothelial-specific transgenic lines () showed a 3-fold decreased caudal vein plexus (CVP) in morphants versus controls at 48 hpf, suggesting a potential role for in angiogenesis. These findings confirm that is essential for angiogenesis, in addition to its involvement in hemostasis.
PubMed: 35386206
DOI: 10.3389/fcell.2022.852989 -
The Science of the Total Environment Nov 2020Surfactants are widely used in the industry of detergents, household products, and cosmetics. SAPDMA is a cationic surfactant that is used mostly in cosmetics,...
Surfactants are widely used in the industry of detergents, household products, and cosmetics. SAPDMA is a cationic surfactant that is used mostly in cosmetics, conditioning agents and has recently gained attention as a corrosion inhibitor in the sea pipelines industry. In this regard, literature concerning the ecotoxicological classification of SAPDMA on aquatic animals is lacking. This study aims to evaluate the potential ecotoxicity of SAPDMA using the aquatic zebrafish embryo model. The potential toxic effects of SAPDMA were assessed by different assays. This includes (i) mortality/survival assay to assess the median lethal concentration (LC); (ii) teratogenicity assay to assess the no observed effect concentration (NOEC); (iii) organ-specific toxicity assays including cardiotoxicity, neurotoxicity (using locomotion assay), hematopoietic toxicity (hemoglobin synthesis using o-dianisidine staining), hepatotoxicity (liver steatosis and yolk retention using Oil Red O (ORO) stain); (iv) cellular cytotoxicity (mitochondrial membrane potential) by measuring the accumulation of JC-1 dye into mitochondria. Exposure of embryos to SAPDMA caused mortality in a dose-dependent manner with a calculated LC of 2.3 mg/L. Thus, based on the LC value and according to the Fish and Wildlife Service (FWS) Acute Toxicity Rating Scale, SAPDMA is classified as "moderately toxic". The No Observed Effect Concentration (NOEC) concerning a set of parameters including scoliosis, changes in body length, yolk, and eye sizes was 0.1 mg/L. At the same NOEC concentration (0.1 mg/L), no organ-specific toxicity was detected in fish treated with SAPDMA, except hepatomegaly with no associated liver dysfunctions. However, higher SAPDMA concentrations (0.8 mg/L) have dramatic effects on zebrafish organ development (eye, heart, and liver development). Our data recommend a re-evaluation of the SAPDMA employment in the industry setting and its strictly monitoring by environmental and public health agencies.
Topics: Animals; Dimethylamines; Embryo, Nonmammalian; Lethal Dose 50; Surface-Active Agents; Water Pollutants, Chemical; Zebrafish
PubMed: 32886985
DOI: 10.1016/j.scitotenv.2020.140450 -
Molecules (Basel, Switzerland) Dec 2023Porous covalent organic frameworks (COFs) have been widely used for the efficient removal of iodine from solution due to their abundance of electron-rich sites. In this...
Porous covalent organic frameworks (COFs) have been widely used for the efficient removal of iodine from solution due to their abundance of electron-rich sites. In this study, two kinds of ketoenamine-based COFs, TpBD-(OMe) and TpBD-Me, are successfully synthesized via Schiff base reaction under solvothermal conditions using 1, 3, 5-triformylphoroglucinol as aldehyde monomer, o-tolidine and o-dianisidine as amino monomers. The ability of TpBD-(OMe) and TpBD-Me to adsorb iodine in cyclohexane or aqueous solutions has been quantitatively analyzed and interpreted in terms of adsorption sites. TpBD-Me possesses two adsorption sites, -NH- and -C=O, and exhibits an adsorption capacity of 681.67 mg/g in cyclohexane, with an initial adsorption rate of 0.6 g/mol/min with respect to COF unit cell. The adsorption capacity of TpBD-(OMe) can be as high as 728.77 mg/g, and the initial adsorption rate of TpBD-(OMe) can reach 1.2 g/mol/min in the presence of oxygen atoms between the methyl group and the benzene ring. Compared with TpBD-Me, the higher adsorption capacity and adsorption rate of TpBD-(OMe) towards iodine are not only reflected in organic solvents, but also in aqueous solutions. It is proven through X-ray photoelectron spectroscopy and Raman spectroscopy that iodine exists in the form of I, I, and I within TpBD-(OMe) and TpBD-Me after adsorption. This work not only expands the application of COFs in the field of iodine adsorption, but also provides research ideas and important an experimental basis for the optimization of iodine adsorption sites.
PubMed: 38138639
DOI: 10.3390/molecules28248151 -
Bioprocess and Biosystems Engineering Mar 2023Dye-contaminated wastewater discharge from textile and dye manufacturing industries is reported as a world worse water polluter due to the toxic and mutagenic behavior...
Dye-contaminated wastewater discharge from textile and dye manufacturing industries is reported as a world worse water polluter due to the toxic and mutagenic behavior of dyes. Peroxidase, one of the key enzymes of oxidoreductases, is widely distributed in nature and has been currently exploited in industries for various applications. Widespread applications of peroxidases are associated with their nonspecific nature towards a wide spectrum of substrates such as phenols, aromatic amines, pesticides, antibiotics, and synthetic dyes. The present study explored the potential of ammonium sulfate precipitated partially purified Brassica oleracea L. var. botrytis leaves peroxidase for degradation of reactive textile dyes Remazol Turquoise Blue 133 G and Drim Red CL4BN. Various physico-chemical parameters such as pH (2-9), temperature (20-70 ℃), enzyme activity (3-24 U/mL), concentrations of HO (0.4-1.4 Mm) and dye (10-100 mg/L) were optimized for enzymatic decolorization of both dyes' solution. Studies revealed that maximum degradation (95%) of Remazol Turquoise Blue 133 G with peroxidase was achieved with 25 mg/L of initial dye concentration, in the presence of 0.8 mM hydrogen peroxide with 45 min of incubation time, at pH 3, 4, and 5, and 70 °C. Maximal decolorization (97%) of Drim Red CL4BN was obtained at pH 2.0, in 10 min of incubation time at 45 ℃ using o-dianisidine hydrochloride as a redox mediator. In conclusion, the findings illustrate the prospect of Brassica oleracea peroxidase to remediate dye pollutants and dye-based industrial effluents in a green technology theme.
Topics: Peroxidase; Botrytis; Hydrogen Peroxide; Peroxidases; Coloring Agents; Textile Industry; Textiles; Plant Leaves; Brassica; Biodegradation, Environmental
PubMed: 36454313
DOI: 10.1007/s00449-022-02820-x -
Methods and Protocols Feb 2020A simple method for the identification of brush-border membrane α-glucosidases is described. The proteins were first solubilized and separated in a gel under native,...
A simple method for the identification of brush-border membrane α-glucosidases is described. The proteins were first solubilized and separated in a gel under native, non-denaturing, conditions. The gel was then incubated in substrate solutions (maltose or sucrose), and the product (glucose) exposed in situ by the oxidation of o-dianisidine, which yields a brown-orange color. Nano-liquid chromatography coupled to mass spectrometry analyses of proteins (nano LC-MS/MS) present in the colored bands excised from the gels, was used to confirm the presence of the enzymes. The stain is inexpensive and the procedure permits testing several substrates in the same gel. Once enzymes are identified, their abundance, relative to that of other proteins in the brush border, can be semi-quantified using nano LC-MS/MS.
PubMed: 32050538
DOI: 10.3390/mps3010015 -
Bio-protocol Jul 2023Myeloperoxidase (MPO) is an enzyme contained in lysosomal azurophilic granules of neutrophils. MPO activity has been shown to correlate with the number of neutrophils in...
Myeloperoxidase (MPO) is an enzyme contained in lysosomal azurophilic granules of neutrophils. MPO activity has been shown to correlate with the number of neutrophils in histological sections of the gastrointestinal tract and is therefore accepted as a biomarker of neutrophil invasion in the gut. This protocol describes an easy, cost-effective kinetic colorimetric assay to quantify myeloperoxidase activity in intestinal tissue samples. It is explained using tissue collected in mice but can also be used for other laboratory animals. In a first step, tissue specimens are homogenized using a phosphate buffer containing 0.5% hexadecyltrimethylammonium bromide (HTAB), which extracts MPO from neutrophils. The obtained supernatant is added to a reagent solution containing o-dianisidine dihydrochloride, which is a peroxidase substrate. Finally, the change in absorption is measured via spectrophotometry and converted to a standardized unit of enzyme activity. The assay is illustrated and compared to a commercially available enzyme-linked immunoassay (ELISA), demonstrating that MPO activity does not necessarily correlate with MPO protein expression in tissue samples. Key features Optimized for use in mice and rats but can also be used for samples of other species. Measures enzymatic activity instead of mRNA or protein expression. Requires a spectrophotometer. Can be performed in duplo using 10 mg of (dry-blotted) gut tissue or more. Graphical overview.
PubMed: 37456337
DOI: 10.21769/BioProtoc.4758