An application of the 2-nitrobenzenesulfenyl method to proteomic profiling of human colorectal carcinoma: A novel approach for biomarker discovery

In the development of novel biomarkers, the proteomic approach is advantageous because using it the cancer-associated proteins can be directly identified. We previously developed a 2-nitrobenzenesulfenyl (NBS) method to improve quantitative proteome analysis. Here, we applied this method to proteomic profiling of colorectal carcinoma (CRC) to identify novel proteins with altered expression in CRC. Each pair of tumor and normal tissue specimens from 12 CRC patients was analyzed, and approximately 5000 NBS-labeled paired peaks were quantified. Peaks with altered signal intensities (>1.5-fold) and occurring frequently in the samples (>70%) were selected, and 128 proteins were identified by MS/MS analyses as differentially expressed proteins in CRC tissues. Many proteins were newly revealed to be CRC related; 30 were reported in earlier studies of CRC. Six proteins that were up-regulated in CRC (ZYX, RAN, RCN1, AHCY, LGALS1, and VIM) were further characterized and validated by Western blot and immunohistochemistry. All six were found to be CRC-localized, either in cancer cells or in stroma cells near the cancer cells. These results indicate that the proteins identified in this study are novel candidates for CRC markers, and that the NBS method is useful in proteome mining to discover novel biomarkers.     

Interannual variations in vegetation activities and climate variability caused by ENSO in tropical rainforests

Interannual variations in terrestrial carbon cycle over tropical rainforests affect the global carbon cycle. Terrestrial ecosystem models show the interannual relationship between climate changes due to El Niño‐Southern Oscillation (ENSO) and net primary production over tropical rainforests. However, we need an independent analysis using satellite‐based vegetation index and climate parameters. In the present study, we extracted the ENSO‐related interannual variations from time‐series in Normalized Difference Vegetation Index (NDVI) and climate data from 1981 to 2000, and analysed their relevance. We detected relationships among NDVI, ENSO, and climate parameters from long‐term data with negative NDVI–ENSO, NDVI–temperature, and positive NDVI–precipitation relations. These correlations suggest that interannual variability in vegetation activities over tropical rainforests could be extracted from NDVI time‐series despite noise components in NDVI data, and that interannual changes in precipitation and temperature caused by ENSO play a more important role in vegetation activities over tropical rainforests than in incoming surface solar radiation.     

Investigation of pathways of advanced glycation end-products accumulation in macrophages

Advanced glycation end-products (AGE) play a role in the pathogenesis of several diseases, including diabetic complications and atherosclerosis. In atherosclerotic lesions of human aortas, AGE are localized in the extracellular matrix and intracellularly in foam cells. Two interpretations are possible for AGE accumulation inside macrophages, one is endocytic uptake of extracellular AGE-proteins by scavenger receptors; the other is intracellular AGE formation inside the macrophages. In the present study, we determined the pathways involved in AGE accumulation inside macrophages. RAW 264.7 cells, a murine macrophage cell line, incubated with BSA and 1600 mM glucose for 40 weeks, recognized heavily modified AGE- BSA. In contrast, the cells showed no ligand activity for mildly modified AGE-BSA, prepared by incubating BSA with 50 mM glucose for 24 weeks. Nepsilon-(carboxymethyl)lysine (CML)-modified proteins of about 65 kDa were detected in human monocyte-derived macrophages incubated for 7 days with 30 mM glucose and phorbol myristate acetate. Furthermore, CML was generated when glycated protein was incubated with hypochloric acid. Taken together, our results indicate that AGE detected inside foam cells in atherosclerotic lesions are generated intracellularly rather than representing endocytic uptake of extracellular AGE-proteins by scavenger receptors.     

Hepatotoxin (microcystin) and neurotoxin (anatoxin-a) contained in natural blooms and strains of cyanobacteria from Japanese freshwaters

Amounts of hepatotoxic microcystin and neurotoxic anatoxin-a were estimated in natural blooms and strains of cyanobacteria from freshwaters in Japan. A simultaneous analysis method of anatoxin-a and microcystin was applied to natural bloom samples, which has been dominated by several species and the strains of cyanobacteria which produced simultaneously both toxins. The natural blooms examined in the present study were mainly composed of Anabaena and Oscillatoria, but most also contained Microcystis and other cyanobacteria. Only one sample was almost unialgal, Anabaena spiroides, collected from Lake Sagami. The toxins in 14 samples collected from nine different natural blooms during 1988-1992 were identified as microcystins-RR, -YR, and -LR; desmethyl-7-microcystin-LR (7-DMLR); and anatoxin-a. Microcystins were the main toxins contained in these natural blooms, with anatoxin-a not being detected or of very little quantity. 7-DMLR was detected in samples only from Lake Kasumigaura. Five strains of Anabaena isolated from waters in Japan produced a small amount of anatoxin-a, but no microcystins. One half of the strains of Microcystis produced microcystins and/or anatoxin-a. This is the first study showing Microcystis producing both anatoxin-a and microcystins.     

A study on a porous residential building model in hot and humid regions: Part 1—the natural ventilation performance and the cooling load reduction effect of the building model

This study targets environmental load reduction in hot and humid regions. It reveals the effects that porous residential buildings have on the natural ventilation performance and, consequently, the cooling load reduction. Two residential building models, namely a model with a void ratio of 0% and a model with a void ratio of 50%, are evaluated using computational fluid dynamics (CFD) analysis and thermal and airflow network analysis. The analysis on components of the heat load indicates that improvements in the natural ventilation performance would significantly reduce the cooling load.     

Simultaneous Perturbation Stochastic Approximation with Norm‐Limited Update Vector

This paper addresses the convergence of simultaneous perturbation stochastic approximation (SPSA) with a norm‐limited update vector. We first illustrate an unstable solution of the standard SPSA algorithm which motivates the consideration of a modified version, where the norm of the update vector is limited to a certain value. Next, a result on the almost‐sure convergence is presented by reducing the modified algorithm into the standard SPSA algorithm and restricting the probability distribution for the perturbation to a Bernoulli distribution. Finally, we apply the modified algorithm to a system identification problem to demonstrate its performance.     

The effect of small dose of topiroxostat on serum uric acid in patients receiving hemodialysis

Introduction : Topiroxostat, a recently developed xanthine oxidase inhibitor, is expected to have fewer adverse effects than allopurinol because it has different mechanism of action from alloprinol. However, its dosage, usage and safety have not been established in patients with impaired renal function or those undergoing dialysis at the development since no studies was conducted in these patients. Methods : Cross over clinical trial using 3 months of allopurinol and topiroxostat on 27 maintain Japanese HD patients were carried out. The effects on oxidative stress status of both drugs were also evaluated by measuring oxidation reduction potential. Findings : Twenty‐five of twenty‐seven patients completed study. The mean serum uric acid levels in the topiroxostat‐treated arm was significantly lower than it in the allopurinol‐treated arm time‐dependently (P < 0.0001). Corrected oxidative stress ratio defined as biological antioxidant potential/diacron reactive oxygen metabolites was significantly increased in topiroxostat‐arm (*P = 0.0035), but not in allopurinol‐arm (P = 0.1429). No significant difference was seen in diacron reactive oxygen metabolites, biological antioxidant potential, static oxidation‐reduction potential, and capacity oxidation‐reduction potential between pre and post treatment of both drugs. Discussion : It is suggested that a low dose of topiroxostat decreased serum uric acid sufficiently to maintain it below 7.0 mg/dL in patients receiving hemodialysis.     

Simultaneous determination of five antioxidants in food by HPLC with fluorescence detection

An HPLC method with fluorescence detection was developed for the determination of propyl gallate, nordihydroguaiaretic acid, butylated hydroxyanisole (2- and 3-tert-butyl-4-hydroxyanisole), tert-butylhydroquinone and octyl gallate in edible oils and foods. The antioxidants in edible oil were isolated directly with acetonitrile saturated with n-hexane. The antioxidants in food were extracted with ethyl acetate and the extract was concentrated under vacuum. They were isolated from the residue with acetonitrile saturated with n-hexane. The acetonitrile layer was centrifuged at 5,000 rpm for 10 min. The HPLC separation was performed on a Symmetry C18 column (3.5 microns, 4.6 mm i.d. x 150 mm) using a mixture of 5% acetic acid-acetonitrile-methanol (4:3:3, v/v/v) as the mobile phase and monitored by using a fluorescence detector with time programming. Sample peaks were identified by comparison of the fluorescence spectra with those of antioxidant standards. Average recoveries of fortified antioxidants at 100 micrograms/g were 72.1-99.6%. Coefficients of variation were 0.7-7.2%.     

The effect of lutein supplementation on visual fatigue: A psychophysiological analysis

We used psychophysiological technology to examine the effect of an oral supplement, a combination of lutein, zeaxanthin and blackcurrant extract (LUT), on visual fatigue, within the context of a randomized, double-blind, placebo-controlled cross-over trial. The LUT supplement and placebo samples were randomly assigned to thirteen participants, who took the samples for two LUT (and vice versa) for another 2 week. Each participant completed visual proof reading tasks for 2 h during each of four testing sessions. Saccade tests were administered before and after the proof reading task, during which the participants moved their eyes back and forth between two targets positioned in the center of two checkerboards. We recorded EEG, EOG, heart rate, and facial muscle potential/performance during the saccade tests. Blood pressure was measured and subjective fatigue and stress scores were collected before and after the proof reading task. We averaged EEG starting at saccade offset in order to analyze eye fixation related potentials (EFRP). Our results suggested that the proof reading task induced visual fatigue. An analysis of EFRP and other psychophysiological data revealed significant differences between the LUT and placebo conditions. These results suggest that supplementation with LUT could help to reduce symptoms of visual fatigue.