Potent Activation of Human but Not Mouse TRPA1 by JT010

Transient receptor potential (TRP) ankyrin repeat 1 (TRPA1), which is involved in inflammatory pain sensation, is activated by endogenous factors, such as intracellular Zn²⁺ and hydrogen peroxide, and by irritant chemical compounds. The synthetic compound JT010 potently and selectively activates human TRPA1 (hTRPA1) among the TRPs. Therefore, JT010 is a useful tool for analyzing TRPA1 functions in biological systems. Here, we show that JT010 is a potent activator of hTRPA1, but not mouse TRPA1 (mTRPA1) in human embryonic kidney (HEK) cells expressing hTRPA1 and mTRPA1. Application of 0.3–100 nM of JT010 to HEK cells with hTRPA1 induced large Ca²⁺ responses. However, in HEK cells with mTRPA1, the response was small. In contrast, both TRPA1s were effectively activated by allyl isothiocyanate (AITC) at 10–100 μM. Similar selective activation of hTRPA1 by JT010 was observed in electrophysiological experiments. Additionally, JT010 activated TRPA1 in human fibroblast-like synoviocytes with inflammation, but not TRPA1 in mouse dorsal root ganglion cells. As cysteine at 621 (C621) of hTRPA1, a critical cysteine for interaction with JT010, is conserved in mTRPA1, we applied JT010 to HEK cells with mutations in mTRPA1, where the different residue of mTRPA1 with tyrosine at 60 (Y60), with histidine at 1023 (H1023), and with asparagine at 1027 (N1027) were substituted with cysteine in hTRPA1. However, these mutants showed low sensitivity to JT010. In contrast, the mutation of hTRPA1 at position 669 from phenylalanine to methionine (F669M), comprising methionine at 670 in mTRPA1 (M670), significantly reduced the response to JT010. Moreover, the double mutant at S669 and M670 of mTRPA1 to S669E and M670F, respectively, induced slight but substantial sensitivity to 30 and 100 nM JT010. Taken together, our findings demonstrate that JT010 potently and selectively activates hTRPA1 but not mTRPA1.     

Macrophage Activation Markers Predict Liver-Related Complications in Primary Biliary Cholangitis

Primary biliary cholangitis (PBC) has a wide variation in clinical presentation and course. There is no significant correlation between these symptoms and the disease stage, although patients with more advanced stages generally have more symptoms. It is important to develop biomarkers in order to identify patients with an increased risk of complications and end-stage liver disease. This study investigated surrogate markers for risk estimation of PBC-related complications, including a study population of 77 patients with PBC who underwent liver biopsy and were measured for serum levels of macrophage activation markers, soluble CD163 (sCD163), soluble mannose receptor (sMR), and zonulin. Patients with PBC were divided into symptomatic (Group S, n = 20) and asymptomatic (Group A, n = 57) groups. The correlations of histological stages based on both Scheuer and Nakanuma classifications with the three serum markers were investigated. The Nakanuma classification involves grading for liver fibrosis and bile duct loss. The three biomarkers were assessed for their diagnostic ability to identify patients with PBC having high risk of developing complications. The predictive factors of these complications were examined as well. Group S had significantly higher serum sMR (p = 0.011) and sCD163 (p = 0.048) levels versus Group A. A composite index of sMR and sCD163 measurements had significantly better prediction performance than sCD163 alone (p = 0.012), although not when compared to sMR alone (p = 0.129). Serum sMR was an independent factor for developing complications on both univariate (Odds ratio (OR) = 30.20, 95% confidence interval (95% CI): 3.410–267.0, p = 0.00220), and multivariate (OR = 33.70, 95% CI: 3.6600–311.0, p = 0.0019) analyses. Patients with PBC having sMR of ≥56.6 had a higher incidence of clinical complications versus those with a sMR of <56.6. Serum sMR predicts the development of complications in patients with PBC. sMR plus sCD163 showed better predictive power than either marker alone, although the addition of sCD163 did not improve the predictive power of sMR. Future prospective studies are required in order to validate the findings of the present study.     

Inhibition of xanthine oxidase activity by an oxathiolanone derivative of quercetin

An oxathiolanone derivative of rutin could be produced in the stomach after the ingestion of rutin containing foods, and the oxathiolanone derivative could be hydrolysed to an oxathiolanone derivative of quercetin (quercetin-oxathiolanone) in the intestine. Quercetin-oxathiolanone as well as quercetin inhibited xanthine oxidase. Approximately 0.05 μM quercetin-oxathiolanone inhibited the activity by 50%, whereas 50% inhibition by quercetin was observed at approximately 0.4 μM. The results suggested that quercetin-oxathiolanone can be used as an effective inhibitor of xanthine oxidase and that the ingestion of rutin-rich foods may be useful to prevent the increase in the blood concentration of uric acid.     

Multiresidue determination of quinolones in animal and fishery products by HPLC

A simple and rapid multiresidue method was developed for the determination of twelve quinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, flumequine, marbofloxacin, nalidixic acid, norfloxacin, ofloxacin, orbifloxacin, oxolinic acid and sarafloxacin) in muscle, liver, chicken eggs, milk, prawn and rainbow trout.The quinolones were extracted from a sample with acetonitrile-water (95 : 5). A fifth part of the filtered extract was diluted with water to keep the acetonitrile ratio at ca. 60%, and passed through a C18 mini-column. The eluate was evaporated to dryness, and the residues were dissolved in methanol-water (30 : 70) for HPLC analysis.The quinolones were separated on a Inertsil ODS-3V column (4.6 mm i.d.x250 mm) with a gradient system of 0.1% phosphoric acid-acetonitrile as the mobile phase, with fluorescence detection.No interfering peak was found on the chromatograms of animal and fishery products, except for milk. The recoveries of the quinolones were over 60% from the animal and fishery products fortified at 0.1 microg/g, and the quantification limits of the quinolones were 0.005 microg/g. This proposed method was found to be effective and suitable for the screening of the quinolones in animal and fishery products.     

A novel gene delivery system in plants with calcium alginate micro-beads

We have produced micrometer-sized calcium alginate beads referred to as "bio-beads" that encapsulate plasmid DNA molecules carrying a reporter gene. In order to evaluate the efficiency of the bio-beads in mediating genetic transfection, protoplasts isolated from cultured tobacco cells (BY-2) were transfected with bio-beads containing a plasmid that carries the modified green fluorescent protein gene CaMV35S-sGFP. With the bio-beads treatment, approximately ten-fold higher GFP expression was observed after 24 h incubation compared to that with the conventional method using a naked plasmid solution. Transfection was up to 0.22% efficient. These results indicate that bio-beads have a possibility for efficient transformation in plants.     

Iterative learning control of Hamiltonian systems: I/O based optimal control approach

In this note, a novel iterative learning control scheme for a class of Hamiltonian control systems is proposed, which is applicable to electromechanical systems. The proposed method has the following distinguished features. This method does not require either the precise knowledge of the model of the target system or the time derivatives of the output signals. Despite the lack of information, the tracking error monotonously decreases in L/sub 2/ sense and, further, perfect tracking is achieved when it is applied to mechanical systems. The self-adjoint related properties of Hamiltonian systems proven in this note play the key role in this learning control. Those properties are also useful for general optimal control. Furthermore, experiments of a robot manipulator demonstrate the effectiveness of the proposed method.     

Computing upper and lower bounds of rotation angles from digital images

Rotations in the discrete plane are important for many applications such as image matching or construction of mosaic images. We suppose that a digital image A is transformed to another digital image B by a rotation. In the discrete plane, there are many angles giving the rotation from A to B, which we call admissible rotation angles from A to B. For such a set of admissible rotation angles, there exist two angles that achieve the lower and the upper bounds. To find those lower and upper bounds, we use hinge angles as used in Nouvel and Rémila [Incremental and transitive discrete rotations, in: R. Reulke, U. Eckardt, B. Flash, U. Knauer, K. Polthier (Eds.), Combinatorial Image Analysis, Lecture Notes in Computer Science, vol. 4040, Springer, Berlin, 2006, pp. 199-213]. A sequence of hinge angles is a set of particular angles determined by a digital image in the sense that any angle between two consecutive hinge angles gives the identical rotation of the digital image. We propose a method for obtaining the lower and the upper bounds of admissible rotation angles using hinge angles from a given Euclidean angle or from a pair of corresponding digital images.     

Dietary CLA and DHA modify skin properties in mice

This study investigated the influence of PUFA on the properties of mouse skin. Mice (3 wk old) were given free access to oils high in linoleic acid, CLA, or DHA for 4 wk. At the end of the experiment, their skins were compared by both biochemical and histological methods. No significant differences in lipid and collagen contents were detected among treatments, although the FA composition in the skin was altered depending upon the FA composition of the supplemented oils. Electron microscopy revealed that the subcutaneous tissue layers in the CLA and DHA groups were significantly thinner than that in the high linoleic acid group, whereas no differences in the thickness of dermis layers were observed among the three groups. These results suggest that skin properties in mice are readily modified by dietary FA sources within 4 wk of dietary oil supplementation.     

Characteristics of polypyrrole chemically synthesized by various oxidizing reagents

Polypyrrole was chemically synthesized using various oxidizing reagents and examined as a positive electrode material. Physical properties, morphologies and electrochemical characteristics of polypyrrole were greatly influenced by the oxidizing reagent used for polymerization. In general, polypyrrole with a smaller particle size and a larger specific surface area showed better discharge performance than that with a larger particle size and a smaller specific surface area. Polypyrrole was also synthesized on various conducting and nonconducting substrates using Fe(C10₄)₃ as an oxidizing reagent. By using PP nonwoven fabric as a substrate material, a high discharge capacity of 72 mA h g^–1 was obtained. Polypyrrole synthesized on only one side of PP nonwoven fabric was able to be used as an electrode.