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Extraction and Chemistry of Rubber Allergens
Published in Robert N. Phalen, Howard I. Maibach, Protective Gloves for Occupational Use, 2023
The ASTM D 6499 titled “Standard Test Method for Immunological Measurement of Antigenic Protein in Hevea Natural Rubber (HNR) and its Products” employs an inhibition Enzyme-Linked ImmunoSorbent Assay (ELISA) which uses a rabbit polyclonal antiserum that reacts with NRL proteins. The NRL extracts and a reference standard NRL antigen are mixed with the antisera in a serial dilution 96 well plate. The extract/antibody mixture is then transferred to an ELISA plate containing immobilized standard NRL antigen. After an incubation period, the ELISA plate is washed, and a secondary horseradish peroxidase-conjugated anti-rabbit IgG is added. Following incubation and washing, hydrogen peroxide and an o-phenylenediamine substrate are added. After a development period, the enzymatic reaction is stopped, and the absorbance of wells containing the glove extracts and latex protein standards are read at 490 nm. The absorbance reading is inversely related to the protein antigen content of the NRL glove extract. An indirect Hev b antigen ELISA (LEAP assay) that uses the same rabbit anti-Hev b polyclonal antibodies and latex antigen standard as the ASTM method has been reported to correlate well (r = 0.89) with ASTM D 6449.16
Eosinophils in a Guinea Pig Model of Allergic Airways Disease
Published in Gerald J. Gleich, A. Barry Kay, Eosinophils in Allergy and Inflammation, 2019
M. G. Campos, M-C. Seminario, J. K. Shute, T. C. Hunt, S. T. Holgate, M. K. Church
Microtiter plates were coated with antibody 8A12 (2 μg/ml in coating buffer) overnight at 4°C. Standard MBP was applied in concentrations ranging from 0.1 to 10,000 ng/ml with overnight incubation at 4°C. Biotinylated antibody 8D12 was added and incubated for 2 h at room temperature, followed by avidin-biotin peroxidase complex for 30 min. Color was developed using O-phenylenediamine dihydrochloride, and absorbance was read at 490 nm after 30 min.
Biodiversity Bioprospection with Respect to Medicinal Plants
Published in Jayanta Kumar Patra, Gitishree Das, Sanjeet Kumar, Hrudayanath Thatoi, Ethnopharmacology and Biodiversity of Medicinal Plants, 2019
Abhishek Kumar Dwivedy, Vipin Kumar Singh, Somenath Das, Anand Kumar Chaudhari, Neha Upadhyay, Akanksha Singh, Archana Singh, Nawal Kishore Dubey
Perturbations in metabolic pathways are associated with the development of cancer and health-threatening diseases (De Flora et al., 1996). Use of plant-based products in the human diet as long practiced in Indian food system may be considered as one of the most effective approaches to avoid cancer. Essential oil of ginger (GEO) has been reported to possess potent antimutagenic activity (Jeena et al., 2014). Essential oil inhibited the mutation induced by sodium azide, extract of tobacco and 4-nitro-o-phenylenediamine in a dose-dependent manner. The study established the stimulatory action of GEO over phase II carcinogen-metabolizing enzymes and suggested its use in chemoprevention. The antimutagenic potential of essential oils from Citrus sinensis and Citrus latifolia was determined against mutation induced by Methyl-N nitro-N-nitrosoguanidine (MNNG) and Ethyl-N-nitro-N-nitrosoguanidine (ENNG) in Salmonella typhimurium TA100 (Toscano-Garibay et al., 2017). Essential oil from both species represented antimutagenic potential against MNNG as well as 2-amino-anthracene (2AA). However, antimutagenic effect against ENNG was shown by Citrus latifolia only. The observed antimutagenic activity was attributed to the presence of components like β-thujene, α-myrcene, R-(+)-limonene, and γ-terpinene. Antimutagenic efficacy of Foeniculum vulgare essential oil against cyclophosphamide (CP) induced genotoxicity in mice was documented by Tripathi et al. (2013). Foeniculum vulgare essential oil was effective in ameliorating the changes induced by CP such as decreased activities of enzymes like superoxide dismutase and catalase. Pretreatment with essential oil resulted in reduced chromosomal abnormality in bone marrow cells suggesting the antimutagenic activity.
IL-38 blockade induces anti-tumor immunity by abrogating tumor-mediated suppression of early immune activation
Published in mAbs, 2023
John P. Dowling, Pavel A. Nikitin, Fang Shen, Halley Shukla, James P. Finn, Nirja Patel, Cezary Swider, Jamie L. Bingaman-Steele, Chris Nicolescu, Eden L Sikorski, Evan J. Greenawalt, Michael J. Morin, Matthew K. Robinson, Karen Lundgren, Benjamin C. Harman
Recombinant full-length human (generated in-house) or mouse IL-38 (Adipogen #AG-40A-0191-C050) was diluted to a final concentration of 0.5 µg/mL in PBS, dispensed into high binding plates (Corning, #9018) and incubated overnight at 4°C. The plate was then washed three times with wash buffer (PBS+0.05% Tween) and blocked with blocking buffer (PBS+2% BSA) at room temperature (RT) for 1 h. Anti-IL-38 antibodies were incubated for 1 h at RT followed by three washes with wash buffer. The horseradish peroxidase (HRP)-conjugated rabbit anti-mouse secondary antibody (Jackson ImmunoResearch, #309-035-082) was incubated at RT for 1 h followed by three washes with wash buffer. O-phenylenediamine dihydrochloride (OPD) substrate (Sigma, #P8287) was incubated for 10–15 min in dark at RT prior to the addition of stop solution (2N sulfuric acid, R&D systems, #DY994). The amount of hu-IL-38 binding to sample/standard was determined by measuring absorbance at 450 nm using an EnSpire plate reader (model number: 2300–0000, PerkinElmer).
Engineering biosafe cisplatin loaded nanostructured lipid carrier: optimisation, synthesis, pharmacokinetics and biodistribution
Published in Journal of Microencapsulation, 2022
Disha Mittal, Archu Singh, Kanchan Kohli, Anita Kamra Verma
Cis and o-phenylenediamine (OPDA) were obtained from Sigma Aldrich. Tween-80, Span 20, Tween-20, Disodium hydrogen phosphate, sodium hydroxide and sodium dihydrogen phosphate were supplied by SD FINE CHEM. Dimethylformamide (DMF) was obtained from Merck & Co. Poloxamer 188 was procured from BASF. Cetyl alcohol, Gelol, GSM, Stearic acid, Compritol® 888 ATO and Precirol® ATO 5 were obtained as a gift sample from Gattefosse. Olive oil, Oleic acid Vitamin E and Soybean oil were purchased from Loba Chemie. Phenazine methosulfate, Nitroblue tetrazolium, Sodium Pyrophodspahte, NADH, NADPH, GSSG were procured from MP biochemicals. 5,5′-dithiobis-2-nitrobenzoic acid (DTNB), Tri-carboxylic Acid (TCA), Dimethyl sulfoxide (DMSO), H2O2, Sodium taurodeoxycholate, Triton-X, Haematoxylin, Eosin, DPX, Xylene were acquired from SRL. Urea, Creatinine, ALT, AST and ALP testing kits were purchased from Erba Manheim. Balb/c mice were purchased from Small Animal Facility (SAF) AIIMS, New Delhi.
Discovery of new VEGFR-2 inhibitors based on bis([1, 2, 4]triazolo)[4,3-a:3',4'-c]quinoxaline derivatives as anticancer agents and apoptosis inducers
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2021
Nawaf A. Alsaif, Mohammed S. Taghour, Mohammed M. Alanazi, Ahmad J. Obaidullah, Abdulrahman A. Al-Mehizia, Manal M. Alanazi, Saleh Aldawas, Alaa Elwan, Hazem Elkady
The general synthetic pathways adopted for the synthesis of the designed compounds are illustrated in Schemes 1–3. Scheme 1 depicts the synthesis of potassium bis[1, 2, 4]triazolo[4,3-a]quinoxaline-4-thiolate 14. Initially, o-phenylenediamine 6 was refluxed with oxalic acid 7 in the presence of 4 N HCl to afford 2,3-(1H,4H)-quinoxalinedione 849. Chlorination of compound 8 was done by refluxing with thionyl chloride yielding 2,3- dichloroquinoxaline 949. Subsequent treatment of the latter with hydrazine hydrate in absolute ethanol afforded 2-chloro-3-hydrazinylquinoxaline 1050. Heating of compound 10 with triethyl orthoformate gave 4-chloro[1, 2, 4]triazolo[4,3-a]quinoxaline 1150. The obtained compound 11 was heated with hydrazine hydrate to afford 4-hydrazinyl-[1, 2, 4] triazolo[4,3-a]quinoxaline 1251. Moreover, reflux of 12 in an alcoholic mixture of carbon disulphide and potassium hydroxide afforded bis[1, 2, 4]triazolo[4,3-a:3′,4′-c]quinoxaline-3-thiol 1344. Heating compound 13 with an alcoholic solution of potassium hydroxide gave the corresponding potassium salt 1444. (Scheme 1)