Anti-neutrophil cytoplasmic antibody-associated (ANCA-associated) small vessel necrotizing vasculitis is caused by immune-mediated inflammation of the vessel wall and is diagnosed in some cases by the presence of myeloperoxidase-specific antibodies (MPO-ANCA). This multicenter study sought to determine whether differences in ANCA epitope specificity explain why, in some cases, conventional serologic assays do not correlate with disease activity, why naturally occurring anti-MPO autoantibodies can exist in disease-free individuals, and why ANCA are undetected in patients with ANCA-negative disease. Autoantibodies from human and murine samples were epitope mapped using a highly sensitive epitope excision/mass spectrometry approach. Data indicated that MPO autoantibodies from healthy individuals had epitope specificities different from those present in ANCA disease. Importantly, this methodology led to the discovery of MPO-ANCA in ANCAnegative disease that reacted against a sole linear sequence. Autoantibodies against this epitope had pathogenic properties, as demonstrated by their capacity to activate neutrophils in vitro and to induce nephritis in mice. The confounder for serological detection of these autoantibodies was the presence of a fragment of ceruloplasmin in serum, which was eliminated in purified IgG, allowing detection. These findings implicate immunodominant epitopes in the pathology of ANCA-associated vasculitis and suggest that autoantibody diversity may be common to other autoimmune diseases.

Background The present study was conducted among Chinese workers employed in glue‐ and shoe‐making factories who had an average daily personal benzene exposure of 31±26 ppm (mean±SD). The metabolites monitored were S‐phenylmercapturic acid (S‐PMA), trans, trans‐muconic acid (t,t‐MA), hydroquinone (HQ), catechol (CAT), 1,2,4‐trihydroxybenzene (benzene triol, BT), and phenol. Methods S‐PMA, t,t‐MA, HQ, CAT, and BT were quantified by HPLC‐tandem mass spectrometry. Phenol was measured by GC‐MS. Results Levels of benzene metabolites (except BT) measured in urine samples collected from exposed workers at the end of workshift were significantly higher than those measured in unexposed subjects (P < 0.0001). The large increases in urinary metabolites from before to after work strongly correlated with benzene exposure. Concentrations of these metabolites in urine samples collected from exposed workers before work were also significantly higher than those from unexposed subjects. The half‐lives of S‐PMA, t,t‐MA, HQ, CAT, and phenol were estimated from a time course study to be 12.8, 13.7, 12.7, 15.0, and 16.3 h, respectively. Conclusions All metabolites, except BT, are good markers for benzene exposure at the observed levels; however, due to their high background, HQ, CAT, and phenol may not distinguish unexposed subjects from workers exposed to benzene at low ambient levels. S‐PMA and t,t‐MA are the most sensitive markers for low level benzene exposure. Am. J. Ind. Med. 37:522–531, 2000. © 2000 Wiley‐Liss, Inc.

An analytical method based on reversed-phase microbore sensitive and highly selective element specific detector. HPLC HPLC coupled on-line with electrospray mass spectrometry coupled on-line with ICP-MS has greatly enhanced our ( ES-MS) is described. The method allows for the capabilities of speciating metals present in a variety of matedetermination of up to ten organoarsenicals in a single rials. 1,[8][9][10][11] This technique provides excellent sensitivity and chromatographic run. Excellent sensitivity and selectivity is detection limits, good selectivity, simplicity and short analysis achieved by operating the triple quadrupole mass spectrometer times. However, the technique has a fundamental limitation, in the selected-reaction monitoring mode. This is possible originating from the fact that identification of metal combecause the ions produced by collision-induced dissociation of pounds is based solely on the matching of retention times. As the protonated molecules or molecular ions of the ten a consequence, errors can result from using this approach, organoarsenicals are characteristic of each compound. The especially in cases of co-eluting metal species. Furthermore, best LODs, achieved in the positive-ion mode, were between 2 lack of appropriate standards could prevent identification and 21 pg of arsenic and corresponded to arsenicals which altogether.12 exist as cations in acidic solutions. The selectivity achieved by Electrospray mass spectrometry (ES-MS) is a relatively new using this method allows for successful determination of technique used with great success for the structural characterizarsenicals co-eluting during HPLC. This is a major ation of numerous compounds of biological and environmental improvement over other hyphenated techniques already used interest.13-15 The fact that ES is an extremely eÂective sample for arsenic speciation, e.g., HPLC-ICP-MS. Furthermore, the introduction technique for mass spectrometry, especially for method was used for the analysis of an undiluted urine SRM non-volatile and/or thermally labile species, has convinced in which arsenobetaine was determined to be present at the researchers to evaluate its potential use for the determination low mg l−1 level.of organometallic compounds, particularly those of tin and arsenic.16-21 For arsenic determinations only three studies have, Keywords: Arsenic ; speciation; microbore high-performance so far, evaluated the usefulness of ES-MS.16, 17, 21 Initially, Siu liquid chromatography ; electrospray mass spectrometry ; et al.16 reported on the atmospheric pressure chemical ioniztriple quadrupole; mass spectrometry-mass spectrometry;ation MS and ES-MS of three arsenic compounds [dimethylcollision-induced dissociation; selected-reaction monitoring arsinic acid ( DMA), arsenobetaine (AsB) and arsenocholine (AsC)] and presented their collisionally-induced dissociation Elemental speciation has become increasingly important in spectra. In later work, Siu et al.17 reported on the development making risk assessments of toxic elemen...

Oxidative stress is known to play important roles in engineered nanomaterial-induced cellular toxicity. However, the proteins and signaling pathways associated with the engineered nanomaterial-mediated oxidative stress and toxicity are largely unknown. To identify these toxicity pathways and networks that are associated with exposure to engineered nanomaterials, an integrated proteomic study was conducted using human bronchial epithelial cells, BEAS-2B and nanoscale titanium dioxide. Utilizing 2-DE and MS, we identified 46 proteins that were altered at protein expression levels. The protein changes detected by 2-DE/MS were verified by functional protein assays. These identified proteins include some key proteins involved in cellular stress response, metabolism, adhesion, cytoskeletal dynamics, cell growth, cell death, and cell signaling. The differentially expressed proteins were mapped using Ingenuity Pathway Analyses™ canonical pathways and Ingenuity Pathway Analyses tox lists to create protein-interacting networks and proteomic pathways. Twenty protein canonical pathways and tox lists were generated, and these pathways were compared to signaling pathways generated from genomic analyses of BEAS-2B cells treated with titanium dioxide. There was a significant overlap in the specific pathways and lists generated from the proteomic and the genomic data. In addition, we also analyzed the phosphorylation profiles of protein kinases in titanium dioxide-treated BEAS-2B cells for a better understanding of upstream signaling pathways in response to the titanium dioxide treatment and the induced oxidative stress. In summary, the present study provides the first protein-interacting network maps and novel insights into the biological responses and potential toxicity and detoxification pathways of titanium dioxide.

Atomic masses and isotopic abundances are independent and complementary properties for discriminating among ion compositions. The number of possible ion compositions is greatly reduced by accurately measuring exact masses of monoisotopic ions and the relative isotopic abundances (RIAs) of the ions greater in mass by +1 Da and +2 Da. When both properties are measured, a mass error limit of 6-10 mDa (< 31 ppm at 320 Da) and an RIA error limit of 10% are generally adequate for determining unique ion compositions for precursor and fragment ions produced from small molecules (less than 320 Da in this study). 'Inherent interferences', i.e., mass peaks seen in the product ion mass spectrum of the monoisotopic [M+H]+ ion of an analyte that are -2, -1, +1, or +2 Da different in mass from monoisotopic fragment ion masses, distort measured RIAs. This problem is overcome using an ion correlation program to compare the numbers of atoms of each element in a precursor ion to the sum of those in each fragment ion and its corresponding neutral loss. Synergy occurs when accurate measurement of only one pair of +1 Da and +2 Da RIAs for the precursor ion or a fragment ion rejects all but one possible ion composition for that ion, thereby indirectly rejecting all but one fragment ion-neutral loss combination for other exact masses. A triple-quadrupole mass spectrometer with accurate mass capability, using atmospheric pressure chemical ionization (APCI), was used to measure masses and RIAs of precursor and fragment ions. Nine chemicals were investigated as simulated unknowns. Mass accuracy and RIA accuracy were sufficient to determine unique compositions for all precursor ions and all but two of 40 fragment ions, and the two corresponding neutral losses. Interrogation of the chemical literature provided between one and three possible compounds for each of the nine analytes. This approach for identifying compounds compensates for the lack of commercial ESI and APCI mass spectral libraries, which precludes making tentative identifications based on spectral matches.