Determination of genotoxic phenylhydrazine agaritine in mushrooms using liquid chromatography-electrospray ionization tandem mass spectrometry

A new method with good sensitivity and specificity for detecting and quantifying genotoxic hydrazines, agaritine and 4-(hydroxymethyl)phenylhydrazine (HMPH), was developed using liquid chromatography-electrospray tandem mass spectrometry (MS). Synthetic agaritine and HMPH were structurally assigned by ¹H-, ¹³C- and two-dimensional nuclear magnetic resonance (NMR) analysis (HMBC and HMQC), high-resolution fast-atom bombardment (HR-FAB) MS and time of flight (TOF) MS. The polar molecule agaritine was separated on an ODS column using 0.01% AcOH-MeOH (99:1, v/v) as an eluent with a simple solid-phase extraction cleanup. There were no interference peaks for any of the mushrooms. Agaricus spp. contained 1247 and 2017 µg g^-1 agaritine. Other species of mushroom had no agaritine. Recoveries of agaritine from spiked mushroom samples were 60.3-114%. Intra-day precision values were 5.5 and 4.2%, and the inter-day precision values were acceptable (15.0 and 23.0%), as agaritine is unstable. The limit of quantification was 0.003 µg g^-1. Even a trace amount of agaritine in mushrooms can, therefore, be determined using this method. We also directly analysed HMPH, an active free hydrazine form of genotoxic agaritine, and obtained direct evidence of its absence from mushrooms. A precursor ion scan confirmed that agaritine derivatives, which could exert similar toxicity, were absent. The results indicate that this specific and sensitive analytical method for detecting and quantifying agaritine and its derivatives could help evaluate the risk of mushroom hydrazines to humans.     

Renal Cyclooxygenase Products are Higher and Lipoxygenase Products are Lower in Early Disease in the pcy Mouse Model of Adolescent Nephronophthisis

Nephronophthisis (NPHP) is a pediatric form of hereditary polycystic kidney disease (PKD), and is the leading cause of end stage renal disease in children. The pcy mouse is an orthologous model of human NPHP, with a mutation in the Nphp3 gene. Renal phospholipase A₂, cyclooxygenase (COX) 1 and cyclic AMP are elevated in this model, suggesting that eicosanoid formation may be altered. In another type of PKD observed in the Han:SPRD-Cy rat, inhibition of eicosanoid production slows disease progression. If renal eicosanoids are similarly altered in NPHP, potential for pharmacologic intervention also may exist for this disorder. Therefore, renal fatty acids and eicosanoids were determined in pcy and normal mice at 15, 30 and 60 days of age by gas chromatography and HPLC–tandem mass spectrometry, respectively. Renal cysts in enlarged kidneys were observed in pcy mice by 15 days of age and increased over time. Renal phospholipid ARA levels were higher in pcy compared to normal mice at 15 and 30 days. Eicosanoid differences were observed starting at 30 days, when the COX products 6-keto-prostaglandin (PG) F_1α, thromboxane B₂ and PGE₂ were higher in pcy compared to normal kidneys. Overall, total COX products were elevated at 30 and 60 days. In contrast, the levels of the lipoxygenase (LOX) products were not altered until 60 days of age and these were lower in pcy kidneys compared to normal. These findings suggest that altered eicosanoids play a role in NPHP, and that manipulating these levels with pharmacologic agents may have therapeutic potential.     

Realization of anti-slip/skid re-adhesion control for electric commuter train based on disturbance observer

Recently, disturbance observer has been used in many system and industry applications. This paper focus on the fine motion control technology based on disturbance observer for electric commuter train. The improvement of adhesion characteristics is important in electric commuter train. We propose the anti-slip/skid re-adhesion control system based on disturbance observer and sensor-less vector control. The effectiveness of the proposed method is confirmed by the experiment based on the actual electric commuter train, which is Series 205-5000 of East Japan Railway Company. Moreover, in order to extend the anti-slip/skid re-adhesion control considering the bogie vibration phenomenon, we propose a new anti-slip re-adhesion control based on the high order disturbance observer considering the resonant frequency of bogie system. Copyright © 2009 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.     

Effect of non-genetically modified (non-GM) soy varieties on the measured value of GM soy by a quantitative PCR method

It is very important to examine the effect of non-genetically modified (non-GM) soy varieties, which constitute the matrix of the testing sample used to quantify GM soy (RRS), on the measured value of RRS by quantitative PCR methods. Therefore, we quantified the amount of RRS in powder-mixed samples containing 1 or 5% RRS prepared by using 10 different varieties of non-GM soy as the matrix. The results revealed that the measured values were not in agreement with the powder-mixing levels and that the extent of the difference depended on the variety of non-GM soy used as the matrix. The yields of DNA extracted differed among the soy varieties. On the other hand, analysis of DNA-mixed samples, that were prepared with the DNAs extracted from RRS and non-GM soy varieties, showed that the measured values of RRS were in agreement with the DNA-mixing levels. These results strongly suggest that the proportions of DNA derived from RRS and non-GM soy were not consistent with the powder-mixing ratio in the case of some non-GM soy varieties used as a matrix, resulting in the discrepancy between the measured values and the powder-mixing levels.     

Laboratory-performance study of the notified methods to detect genetically modified maize (CBH351) and potato (NewLeaf Plus and NewLeaf Y)

To investigate the key factors affecting the reliability of the analytical results, a laboratory-performance study was attempted for the notified methods to detect genetically modified (GM) maize (CBH351) and GM potato (NewLeaf Plus and NewLeaf Y). The test samples were designed as three pairs of blind duplicates, which included 0%, 0.1% and 1.0% GM maize (CBH351) or GM potato (NewLeaf Plus or NewLeaf Y). Fourteen laboratories participated in the study. The test samples were sent to the participating laboratories along with the protocol. The data were collected from all laboratories and statistically analyzed. For the 0% sample of the CBH351 maize, one laboratory reported a false-positive result. It was considered that contamination could have occurred via the common use of equipment or tools for the test. For the 0.1% samples of the NewLeaf Plus potato or NewLeaf Y potato, on the other hand, three laboratories reported false-negative results. It was presumed that these results were due to changes of the conditions of the electrophoresis and agarose-gel staining. The other laboratories reported appropriate results. It was considered that the method employed in this study was suitable for the assessment of laboratory performance.     

Influences of metal ions on the formation of γ-FeOOH and magnetite rusts

To elucidate the role of anti-corroding alloying metals in weathering steels, the γ-FeOOH and Fe₃O₄ particles were prepared by air-oxidation of FeSO₄ solutions containing different amount of Ti(IV), Cr(III), Cu(II) and Ni(II). The crystallinity and particle size of γ-FeOOH were lowered by adding Cr(III), Cu(II) and Ni(II) but not influenced by Ti(IV). All the metal ions impeded the crystallization and particle growth of Fe₃O₄ in a similar manner. The results on γ-FeOOH and Fe₃O₄ were compared with those on α- and β-FeOOHs previously reported to know the comprehensive effects of metal ions on the formation of rusts.     

Electronic structure of boron-doped diamond with B–H complex and B pair

The electronic structure of boron–hydrogen complex and boron pair in diamond are studied by first-principles density-functional calculations with supercell models. The electronic structure calculated for the B–H complexes with C_2v or C_3v symmetry and the nearest-neighbor B pair is used to interpret recent experimental results such as B 1s x-ray photoemission spectroscopy, ¹¹B nuclear quadruple resonance and B K-edge x-ray absorption spectroscopy, which cannot be explained solely by the isolated substitutional boron.