کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2568243 1128430 2015 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A fluorescence high throughput screening method for the detection of reactive electrophiles as potential skin sensitizers
ترجمه فارسی عنوان
روش غربالگری کارایی فلورسانس برای تشخیص الکتروفیل های واکنشی به عنوان حساسیت کننده های پوستی بالقوه
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم محیط زیست بهداشت، سم شناسی و جهش زایی
چکیده انگلیسی


• A novel fluorescence-based method to detect electrophilic sensitizers is proposed.
• A model fluorescent thiol was used to directly quantify the reaction products.
• A discussion of the reaction workflow and critical parameters is presented.
• The method could provide a useful tool to complement existing chemical assays.

Skin sensitization is an important toxicological end-point in the risk assessment of chemical allergens. Because of the complexity of the biological mechanisms associated with skin sensitization, integrated approaches combining different chemical, biological and in silico methods are recommended to replace conventional animal tests. Chemical methods are intended to characterize the potential of a sensitizer to induce earlier molecular initiating events. The presence of an electrophilic mechanistic domain is considered one of the essential chemical features to covalently bind to the biological target and induce further haptenation processes. Current in chemico assays rely on the quantification of unreacted model nucleophiles after incubation with the candidate sensitizer. In the current study, a new fluorescence-based method, ‘HTS-DCYA assay’, is proposed. The assay aims at the identification of reactive electrophiles based on their chemical reactivity toward a model fluorescent thiol. The reaction workflow enabled the development of a High Throughput Screening (HTS) method to directly quantify the reaction adducts. The reaction conditions have been optimized to minimize solubility issues, oxidative side reactions and increase the throughput of the assay while minimizing the reaction time, which are common issues with existing methods. Thirty-six chemicals previously classified with LLNA, DPRA or KeratinoSens™ were tested as a proof of concept. Preliminary results gave an estimated 82% accuracy, 78% sensitivity, 90% specificity, comparable to other in chemico methods such as Cys-DPRA. In addition to validated chemicals, six natural products were analyzed and a prediction of their sensitization potential is presented for the first time.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Toxicology and Applied Pharmacology - Volume 289, Issue 2, 1 December 2015, Pages 177–184
نویسندگان
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