Mass Spectrometric Analyses of Organophosphate Insecticide Oxon Protein Adducts

OBJECTIVE: Organophosphate (OP) insecticides continue to be used to control insect pests. Acute and chronic exposures to OP insecticides have been documented-to cause adverse health effects, but few OP-adducted proteins have been correlated with these illnesses at the molecular level. Our aim was to review the literature covering the current state of the art in mass spectrometry (MS) used to identify OP protein biomarkers. DATA SOURCES AND EXTRACTION: We identified general and specific research reports related to OP insecticides, OP toxicity, OP structure, and protein MS by searching PubMed and Chemical Abstracts for articles published before December 2008. DATA SYNTHESIS: A number of OP-based insecticides share common structural elements that result in predictable OP-protein adducts. The resultant OP-protein adducts show an increase in molecular mass that can be identified by MS and correlated with the OP agent. Customized OP-containing probes have also been used to tag and identify protein targets that can be identified by MS. CONCLUSIONS: MS is a useful and emerging tool for the identification of proteins that are modified by activated organophosphate insecticides. MS can characterize the structure of the OP adduct and also the specific amino acid residue that forms the key bond with the OP. Each protein that is modified in a unique way by an OP represents a unique molecular biomarker that with further research can lead to new correlations with exposure

Distinct nonequilibrium plasma chemistry of C2 affecting the synthesis of nanodiamond thin films from C2H2 (1%)/H2/Ar-rich plasmas

6 pages, 5 figures, 6 tables.We show that the concentrations of the species C2 (X 1Σg+), C2 (a 3Πu), and C2H exhibit a significant increase when the argon content grows up to 95% in medium pressure (0.75 Torr) radio frequency (rf) (13.56 MHz) produced C2H2 (1%)/H2/Ar plasmas of interest for the synthesis of nanodiamond thin films within plasma enhanced chemical vapor deposition devices. In contrast, the concentrations of CH3 and C2H2 remain practically constant. The latter results have been obtained with an improved quasianalytic space–time-averaged kinetic model that, in addition, has allowed us to identify and quantify the relative importance of the different underlying mechanisms driving the nonequilibrium plasma chemistry of C2. The results presented here are in agreement with recent experimental results from rf CH4/H2/Ar-rich plasmas and suggest that the growth of nanodiamond thin films from hydrocarbon/Ar-rich plasmas is very sensitive to the contribution of C2 and C2H species from the plasma.This work was partially funded by CICYT (Spain) under a Ramón y Cajal project and under Project No. TIC2002- 03235. One of the authors (F.J.G.V.) acknowledges a Ramón y Cajal contract from the Spanish Ministry of Science and Technology (MCYT). One of the authors (J.M.A.) acknowledges partial support from CICYT (Spain) under Project No. MAT 2002-04085-C02-02.Peer reviewe

Eukaryotic Y-family polymerases bypass a 3-methyl-2′-deoxyadenosine analog in vitro and methyl methanesulfonate-induced DNA damage in vivo

N3-methyl-adenine (3MeA) is the major cytotoxic lesion formed in DNA by SN2 methylating agents. The lesion presumably blocks progression of cellular replicases because the N3-methyl group hinders interactions between the polymerase and the minor groove of DNA. However, this hypothesis has yet to be rigorously proven, as 3MeA is intrinsically unstable and is converted to an abasic site, which itself is a blocking lesion. To circumvent these problems, we have chemically synthesized a 3-deaza analog of 3MeA (3dMeA) as a stable phosphoramidite and have incorporated the analog into synthetic oligonucleotides that have been used in vitro as templates for DNA replication. As expected, the 3dMeA lesion blocked both human DNA polymerases α and δ. In contrast, human polymerases η, ι and κ, as well as Saccharomyces cerevisiae polη were able to bypass the lesion, albeit with varying efficiencies and accuracy. To confirm the physiological relevance of our findings, we show that in S. cerevisiae lacking Mag1-dependent 3MeA repair, polη (Rad30) contributes to the survival of cells exposed to methyl methanesulfonate (MMS) and in the absence of Mag1, Rad30 and Rev3, human polymerases η, ι and κ are capable of restoring MMS-resistance to the normally MMS-sensitive strain

Treatable childhood neuronopathy caused by mutations in riboflavin transporter RFVT2.

Childhood onset motor neuron diseases or neuronopathies are a clinically heterogeneous group of disorders. A particularly severe subgroup first described in 1894, and subsequently called Brown-Vialetto-Van Laere syndrome, is characterized by progressive pontobulbar palsy, sensorineural hearing loss and respiratory insufficiency. There has been no treatment for this progressive neurodegenerative disorder, which leads to respiratory failure and usually death during childhood. We recently reported the identification of SLC52A2, encoding riboflavin transporter RFVT2, as a new causative gene for Brown-Vialetto-Van Laere syndrome. We used both exome and Sanger sequencing to identify SLC52A2 mutations in patients presenting with cranial neuropathies and sensorimotor neuropathy with or without respiratory insufficiency. We undertook clinical, neurophysiological and biochemical characterization of patients with mutations in SLC52A2, functionally analysed the most prevalent mutations and initiated a regimen of high-dose oral riboflavin. We identified 18 patients from 13 families with compound heterozygous or homozygous mutations in SLC52A2. Affected individuals share a core phenotype of rapidly progressive axonal sensorimotor neuropathy (manifesting with sensory ataxia, severe weakness of the upper limbs and axial muscles with distinctly preserved strength of the lower limbs), hearing loss, optic atrophy and respiratory insufficiency. We demonstrate that SLC52A2 mutations cause reduced riboflavin uptake and reduced riboflavin transporter protein expression, and we report the response to high-dose oral riboflavin therapy in patients with SLC52A2 mutations, including significant and sustained clinical and biochemical improvements in two patients and preliminary clinical response data in 13 patients with associated biochemical improvements in 10 patients. The clinical and biochemical responses of this SLC52A2-specific cohort suggest that riboflavin supplementation can ameliorate the progression of this neurodegenerative condition, particularly when initiated soon after the onset of symptoms

A Historical Review of Impressed Current Cathodic Protection of Steel in Concrete

This paper reviews the history of the development of impressed current cathodic protection of atmospherically exposed reinforced concrete from the first trials in 1959 on bridges to recently installed systems on a wide range of structures around the world. The paper covers the research efforts, anode developments, control systems and monitoring sensors which are reviewed and their evolution explained. The research into the potential and actual side effects of cathodic protection currents in concrete are summarised. The development of standards and guidance on impressed current cathodic protection is also reviewed