Profiling and Imaging of Phospholipids in Brains of Abcd1‐Deficient Mice

ABCD1 is a gene responsible for X‐linked adrenoleukodystrophy (X‐ALD), and is critical for the transport of very long‐chain fatty acids (VLCFA) into peroxisomes and subsequent β‐oxidation. VLCFA‐containing lipids accumulate in X‐ALD patients, although the effect of ABCD1‐deficiency on each lipid species in the central nervous system has not been fully characterized. In this study, each phospholipid and lysophospholipid species in Abcd1‐deficient mice brains were profiled by liquid chromatography‐mass spectrometry. Among the phospholipid and lysophospholipid species that are significantly more enriched in Abcd1‐deficient mice brains, VLCFA were present in 75, 15, 5, 4, and 1 species of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, lysophosphatidylcholine, and lysophosphatidylethanolamine, respectively. Most VLCFA were incorporated at the sn‐1 position of phosphatidylcholine and phosphatidylethanolamine. Among the phospholipid species that are significantly less enriched in Abcd1‐deficient mice brains, odd‐numbered saturated or mono‐unsaturated fatty acyl moieties are contained in all phosphatidylcholine species. In addition, a number of phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine species contained highly unsaturated fatty acyl moieties. Intriguingly, 44:1 phosphatidylcholine with VLCFA was mainly distributed in the gray matter, such as the cortex, but not in the white matter in the cerebrum and cerebellum. These results show that ABCD1‐deficiency causes metabolic alternation of long‐chain fatty acids and VLCFA. Moreover, our results imply a molecular mechanism for the incorporation of saturated or monounsaturated VLCFA into the sn‐1 position of phospholipids, and also indicate that the distribution of phospholipids with VLCFA may correlate with the development of X‐ALD.     

Simultaneous design of the sensory morphology and controller of mobile robots

This paper proposes a method for automatic design of the sensory morphology of a mobile robot. The proposed method employs two types of adaptations, ontogenetic and phylogenetic, to optimize the sensory morphology of the robot. In ontogenetic adaptation, reinforcement learning searches for the optimal policy, which is highly dependent on the sensory morphology. In phylogenetic adaptation, a genetic algorithm is used to select morphologies with which the robot can learn tasks faster. Our proposed method was applied to the design of the sensory morphology of a line‐following robot. We performed simulation experiments to compare the design solution with a hand‐coded robot. The results of the experiments revealed that our robot outperformed the hand‐coded robot in terms of the following accuracy and learning speed, although our robot had fewer sensors than the hand‐coded one. We also built a physical robot using the design solution. The experimental results revealed that this physical robot used its morphology effectively and outperformed the hand‐coded robot. © 2010 Wiley Periodicals, Inc. Electr Eng Jpn, 172(1): 48–57, 2010; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20965     

Denatured-jacalin derivatives with selective recognition for O-linked glycosides (ST,T, Tn,and STn Type) on IgA1 hinge region

Immunoglobulin A1 (IgA1) concentration in the plasma of patients with IgA nephropathy (IgAN) as the cause of renal failure is higher than that in the plasma of normal controls. IgA1 with abnormal sugars is considered to deposit in the glomerular mesangium, aggravating nephritis in IgAN. Jacalin is a lectin that recognizes sugars on IgA1. However, its selective-recognition for normal-type (ST type, NeuAc-α(2,3)-Gal-β(1,3)-GalNAc) and abnormal-type (T type, Gal-β(1,3)-GalNAc; Tn type, GalNAc; STn type, NeuAc-α(2,6)-GalNAc) sugars α-O-linked to serine/threonine in IgA1 is weak. Therefore, jacalin cannot be used for recognizing specific sugar types on IgA1. We attempted to develop a new recognition method for specific sugar types on IgA1 by utilizing the multirecognition capability of jacalin. Its binding abilities were regulated by heat denaturation with suitable template sugar (galactose or N-acetylgalactosamine). Further, we successfully prepared denatured-jacalin derivatives, which recognized ST-/T-type sugars on IgA1, by sugar-immobilized affinity chromatography. Enzyme-linked immunosorbent assay of denatured-jacalin derivatives, showed the ratios of abnormal sugars on IgA1 in the plasma of IgAN patients and normal controls to be approximately 60% and 20%, respectively. The results proved that profiling of sugar types in IgAN can successfully be performed by solely using jacalin derivatives.     

Correlation of the structures of agricultural fungicides to gene expression in Saccharomyces cerevisiae upon exposure to toxic doses

Correlations between the chemical structures of agricultural fungicides and mRNA expression levels following exposure of Saccharomyces cerevisiae to toxic doses of thiuram, zineb, maneb, TPN, and PCP were examined. Structurally, thiuram, zineb, and maneb are dithiocarbamate fungicides, whereas TPN and PCP are not. To characterize chemical toxicity, genes expression was classified according to the functional groups used by the MIPS database. However, no correlations between the classification scheme and chemical structures were found. Hierarchical clustering of gene expression profiles was performed to characterize the effects of the five chemicals. According to this analysis the similarity of gene expression profiles depended on the similarity of chemical structures. These results suggest that DNA microarray technology has potential for predicting the major chemicals which will cause environmental toxicity and will provide information on new biomonitoring methods.     

Determination of benzimidazole anthelmintics in livestock foods by HPLC

A simple and rapid liquid chromatographic method was developed for the simultaneous determination of twelve benzimidazole anthelmintics in livestock foods using reversed-phase high-performance liquid chromatography with photodiode array detection (PDA). A sample was homogenized with acetonitrile and n-hexane, and centrifuged. The acetonitrile phase was isolated and evaporated. The residue was dissolved in 0.1 mol/L carbonate buffer solution (pH = 9.1), sonicated, and then subjected to clean-up on a Bond Elut LRC-C18 cartridge. The benzimidazole compounds were separated isocratically on a Capcell Pak C18 UG 120 (5 microns, 150 x 4.6 mm i.d.) column and detected by PDA at 295 and 313 nm. Mixtures of acetonitrile and 0.05 mol/L ammonium acetate in mixing ratios of (20:80) and (40:60) were used as the mobile phase, and the flow-rate was 1.0 mL/min at 40 degrees C. The mean recoveries (n = 3) from 0.1-0.5 microgram/g added samples were 72.6-97.2% with coefficients of variation of 0.3-8.5%. The detection limits were 0.01-0.05 microgram/g.     

Affinity capture and elution/electrospray ionization mass spectrometry assay of phosphomannomutase and phosphomannose isomerase for the multiplex analysis of congenital disorders of glycosylation types Ia and Ib

We report a new application of affinity capture-elution electrospray mass spectrometry (ACESI-MS) to assay the enzymes phosphomannomutase (PMM) and phosphomannose isomerase (PMI), which when deficient cause congenital disorders of glycosylation CDG-type Ia and type Ib, respectively. The novel feature of this mass-spectrometry-based assay is that it allows one to distinguish and quantify enzymatic products that are isomeric with their substrates that are present simultaneously in complex mixtures, such as cultured human cell homogenates. This is achieved by coupled assays in which the PMM and PMI primary products are in vitro subjected to another enzymatic reaction with yeast transketolase that changes the mass of the products to be detected by mass spectrometry. The affinity purification procedure is fully automated, and the mass spectrometric analysis is multiplexed in a fashion that is suitable for high-throughput applications.     

Acid-dyeable acrylic fibres copolymerised with amino alkyl methacrylates for differential cross-dyeing

The equilibrium adsorption of acid dyes on acrylic fibres copolymerised with dimethyl amino ethyl methacrylate has been investigated. A typical characteristic of this type of fibre is its excellent stable dyeability over a wide pH range. This is due to the strong basicity of alkylamino groups in the copolymerised component, which makes it possible to dye the fibre thoroughly even in a weakly acidic dyebath using a normal dyeing processes of acrylic fibre with cationic dye. Consequently it is useful for differential cross-dyeing in a single dyebath.