Metabolism and Toxicity of Thioacetamide and Thioacetamide SOxide in Rat Hepatocytes

“This document is the Accepted Manuscript version of a Published Work that appeared in final form in Chemical Research in Toxicology, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/tx3002719The hepatotoxicity of thioacetamide (TA) has been known since 1948. In rats, single doses cause centrilobular necrosis accompanied by increases in plasma transaminases and bilirubin. To elicit these effects TA requires oxidative bioactivation leading first to its S-oxide (TASO) and then to its chemically reactive S,S-dioxide (TASO2) which ultimately modifies amine-lipids and proteins. To generate a suite of liver proteins adducted by TA metabolites for proteomic analysis, and to reduce the need for both animals and labeled compounds, we treated isolated hepatocytes directly with TA. Surprisingly, TA was not toxic at concentrations up to 50 mM for 40 hr. On the other hand, TASO was highly toxic to isolated hepatocytes as indicated by LDH release, cellular morphology and vital staining with Hoechst 33342/propidium iodide. TASO toxicity was partially blocked by the CYP2E1 inhibitors diallyl sulfide and 4-methylpyrazole, and was strongly inhibited by TA. Significantly, we found that hepatocytes produce TA from TASO relatively efficiently by back-reduction. The covalent binding of [14C]-TASO is inhibited by unlabeled TA which acts as a “cold-trap” for [14C]-TA and prevents its re-oxidation to [14C]-TASO. This in turn increases the net consumption of [14C]-TASO despite the fact that its oxidation to TASO2 is inhibited. The potent inhibition of TASO oxidation by TA, coupled with the back-reduction of TASO and its futile redox cycling with TA may help explain phenomena previously interpreted as “saturation toxicokinetics” in the in vivo metabolism and toxicity of TA and TASO. The improved understanding of the metabolism and covalent binding of TA and TASO facilitates the use of hepatocytes to prepare protein adducts for target protein identification

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Identification of a need or a deficiency triggers conceptual system design. The first step in conceptual design is to analyze and translate the need or deficiency into specific qualitative and quantitative customer and design requirements. Design methods such as Quality Function Deployment (QFD), Parameter Taxonomies, and Input/Output Matrices (IOM) provide a useful framework for this translation. Well defined and unambiguous requirements enhance communications and can potentially reduce the number of “detours ” during subsequent design and development phases. However, imprecision and vagueness characterize the conceptual design phase. To accommodate imprecision, the QFD method and the concept selection methodology, initially proposed by Pugh [91], have been modified and extended by applying concepts from fuzzy set theory [Verma and Fabrycky, 1995; and Verma and Knezevic, 1996]. The extended approach provides a rigorous yet graceful mechanism for dealing with imprecise requirements, priorities, and correlations as prerequisites to concept selection. This technical paper presents an expert system based extension to the fuzzy QFD methodology. Emphasis is on the: a) identification of strategic market and product opportunities, b) identification of applied research focus areas, and c) isolation of inconsistencies between customer articulation o

An overview on applications of optical spectroscopy in cervical cancers

Despite advances in the treatment modalities, cervical cancers are one of the leading causes of cancer death among women. Pap smear and colposcopy are the existing screening methods and histopathology is the gold standard for diagnosis. However, these methods have been shown to be prone to reporting errors, which could be due to their subjective interpretation. Radiotherapy is the mainstay of treatment for the locally advanced stages of cervical cancers. The typical treatment regimen spans over 4 months, from the first fraction of radiation to clinical assessment of tumor response to radiotherapy. It is often noticed that due to intrinsic properties of tumors, patients with the same clinical stage and histological type respond differently to radiotherapy. Hence, there exists a need for the development of new methods for early diagnosis as well as for early prediction of tumor radioresponse. Optical spectroscopic methods have been shown to be potential alternatives for use in cancer diagnosis. In this review, we provide a brief background on the anatomy and histology of the uterine cervix and the etiology of cervical cancers; we briefly discuss the optical spectroscopic approach to cervical cancer diagnosis. A very brief discussion on radiation therapy and radiation resistance is also provided. We also share our experiences with the Raman spectroscopic methodologies in cervical cancer diagnosis as well as in the prediction of tumor radioresponse