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Approaches for Identification and Validation of Antimicrobial Compounds of Plant Origin: A Long Way from the Field to the Market
Published in Mahendra Rai, Chistiane M. Feitosa, Eco-Friendly Biobased Products Used in Microbial Diseases, 2022
Lívia Maria Batista Vilela, Carlos André dos Santos-Silva, Ricardo Salas Roldan-Filho, Pollyanna Michelle da Silva, Marx de Oliveira Lima, José Rafael da Silva Araújo, Wilson Dias de Oliveira, Suyane de Deus e Melo, Madson Allan de Luna Aragão, Thiago Henrique Napoleão, Patrícia Maria Guedes Paiva, Ana Christina Brasileiro-Vidal, Ana Maria Benko-Iseppon
The microdilution test is performed on flat-bottomed 96-well microtiter plates and each row of a microplate corresponds to a line. Successive dilutions (1:1) of the antimicrobial agent are performed in the culture medium. Then the same volume of inoculum is added. A well corresponding to 100% growth is prepared without the presence of the antimicrobial agent and a sterility control that has the culture medium and the antimicrobial agent. Then the plates should be incubated in the same conditions as described for the macrodilution test. Dissolved Oxygen (DO) should be measured at the beginning (zero time) and the end of the test using a microplate reader. The IMC will be determined as the lowest concentration of the sample capable of promoting a certain minimum value of OD reduction compared to the growth control. This value can be 50 or 90%, for example.
Determination of Antiviral Activity
Published in Adorjan Aszalos, Modern Analysis of Antibiotics, 2020
The quantity of the test antibiotic available for antiviral testing will seriously influence the kind and quantity of antiviral testing being planned: researchers have tended to use those systems requiring the minimum quantity of test substance. This usually involves the use of disposable 96 well microplates for in vitro experiments [10] and mice for initial in vivo experiments.
Buriti (Mauritia flexuosa L.) Oil Supplementation: Effects on Oxidative Stress and Hormonal Concentrations in Male Wistar Rats
Published in Megh R. Goyal, Hafiz Ansar Rasul Suleria, Ademola Olabode Ayeleso, T. Jesse Joel, Sujogya Kumar Panda, The Therapeutic Properties of Medicinal Plants, 2019
Boitumelo Rosemary Mosito, Nicole Lisa Brooks, Yapo Guillaume Aboua
The BCA assay was used. Five albumin standards of different concentrations were prepared, and 25µL of each standard or sample was added in duplicate to a microplate well. The 200µL of working reagent was added to each well and mixed thoroughly on a plate shaker for 30 seconds then incubated at 37°C for 30 minutes. The plate was cooled at room temperature, and absorbance was measured at 562 nm on a plate reader (Thermo Electron Corporation, Multiskan spectrum, USA). The protein concentrations were quantified by using the standard curve and expressed as µg/ml.
Short-term administration of tibolone reduces inflammation and oxidative stress in the hippocampus of ovariectomized rats fed high-fat and high-fructose
Published in Nutritional Neuroscience, 2023
Norma A. Estrada-Cruz, Leticia Manuel-Apolinar, Julia J. Segura-Uribe, Julio C. Almanza-Pérez, Ángeles Fortis-Barrera, Sandra Orozco-Suárez, Guadalupe Bautista-Poblet, Angélica Coyoy-Salgado, Christian Guerra-Araiza
We determined superoxide dismutase (SOD) activity with a commercial kit (CAYMAN Chemical Company, Michigan, USA) that allows detection of Cu/Zn-, Mn-, and Fe-SOD. The reaction was performed in the wells of microplates. It consisted of the following: 200 μL of tetrazolium salt (radical detector) were dissolved in 19.95 mL of 50 mM Tris-HCl buffer pH 8.0, 0.1 mM diethylenetriaminepentaacetic acid (DTPA), and 0.1 mM hypoxanthine. A volume (10 μL) of either the enzymatic extract or a standard of SOD was added. The reaction began with the addition of 20 μL xanthine oxidase—prepared as a dilution of 50 μL xanthine oxidase in 1.95 mL of 50 mM Tris-HCl, pH 8.0. The tetrazolium salt then captured the superoxide radicals, and we quantified them at 450 nm. Radical concentrations were inversely proportional to SOD activity in the samples.
Antimicrobial activity of flavonoids glycosides and pyrrolizidine alkaloids from propolis of Scaptotrigona aff. postica
Published in Toxin Reviews, 2023
T. M. Cantero, P. I. Silva Junior, G. Negri, R. M. Nascimento, R. Z. Mendonça
To determine the MIC, the technique of inhibiting microbial growth in liquid medium was used with modifications (Riciluca et al. 2012). The test was carried out in 96-well microplates with a final volume of 100μL. The samples were evaluated at concentrations of 31.3 to 200μg/mL, in the culture medium containing 105 CFU/mL for bacteria and/or 104 CFU/mL for fungi. The growth media used were PB (Peptones 10g/L; NaCl 5g/L; pH 7.4) for bacteria and PDB (Potato Dextrose Broth; 1.2g Potato dextrose; 100ml of H2O; pH 5.0) for fungi. In this experiment, 10mg/mL positive control and 20µL ultrapure water (Direct-Q 5UV) were used as a negative control. A solvent control, for all microorganisms, was performed with the maximum concentration of DMSO used to solubilize the samples from the MEP (0.5%). The experiments were carried out in triplicate. Culture growth controls were carried out, as well as appropriate sterility controls of the materials, adding 100µL of ultrapure water (Direct-Q 5UV) and 100µL of the culture media. Microbial growth was evaluated by turbidity of the medium by absorbance measurement (λ: 595nm) in a Victor³ microplate reader (1420 Multilabel Counter/Victor³ – Perkin Elmer), after 24h of incubation at 37° C.
Biodegradable gemcitabine-loaded microdevice with sustained local drug delivery and improved tumor recurrence inhibition abilities for postoperative pancreatic tumor treatment
Published in Drug Delivery, 2022
Xiangming Kong, Miao Feng, Lihuang Wu, Yiyan He, Hongli Mao, Zhongwei Gu
The nuclear magnetic resonance (NMR) spectra of all materials was recorded with an NMR spectrometer (AVANCE III HD 600 MHz, Bruker, Rheinstetten, Germany). The molecular weights of poly(l-lactic-co-glycolic acid) (P(L)LGA) were measured by Gel Permeation Chromatography (GPC, Breeze2, Waters, Milford, MA). The rheological curves of P(L)LGA, PEG, and GEM·HCl mixtures were determined by a rheometer (DHR-2, TA, New Castle, DE). The electrospray ionization (ESI) mass spectrum of GEM·HCl was determined by a mass spectrometer (QExactive, Thermo Fisher Scientific, Waltham, MA). The drug loading (DL) and drug release behavior of all microdevices were measured by a microplate reader (Nivo, PerkinElmer, Waltham, MA). The surface and section morphology of microdevices were observed by a scanning electron microscope (SEM, JSM-IT200, Japan Electronics, Minato, Japan). All histological sections of mice were recorded with inverted fluorescence microscope (IFM, TE2000, Nikon, Minato, Japan). Blood routine tests were measured by an automatic hematology analyzer (BC-2800vet, Mindray, Shenzhen, China). Blood biochemical parameters were determined by an automatic biochemical analyzer (Chemray 800, Rayto, Shenzhen, China).