Purification and characterisation of chicken liver monomeric glutathione peroxidase

1. A novel glutathione peroxidase, which is distinct from tetrameric glutathione peroxidase, was purified to homogeneity from a broiler chick liver cytosolic fraction using 5 different column chromatographic methods.

2. The enzyme in cytosol was separated from ‘classic’ tetrameric glutathione peroxidase and glutathione S‐transferases by DEAE‐Sephacel and Sephadex G‐100 chromatographies and further purified by Mono Q, hydroxylapatite and sulphobro‐mophthalein‐S‐glutathione‐agarose chromatographies.

3. The molecular weight of the purified enzyme determined by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis was 19,500 and that found by gel filtration chromatography was comparable. This indicates that the enzyme protein is a single polypeptide. The isoelectric point of the enzyme was determined as 7.0 by polyacrylamide gel isoelectric focusing.

4. The purified enzyme catalysed the reduction of hydrogen peroxide, cumene hydroperoxide, tert‐butyl hydroperoxide and linoleic acid hydroperoxide. Furthermore, it reduced phosphatidylcholine hydroperoxide in the absence of phospholi‐pase A2. The optimum pH for the enzyme reaction was 7.0. The antiserum against the purified enzyme reacted with the 19.5 kDa polypeptide in the liver cytosol of duck and quail.     

Suppression of Verticillium Wilt in Eggplant by Some Fungal Root Endophytes

One hundred and twenty-three fungal isolates were obtained from 225 root segments of eggplants, melon, tomato, strawberry and Chinese cabbage, grown as bait plants in a mixed soil made up of samples from different fields in Shizuoka, Japan. Isolates belonging to Mycelium radicis atrovirens (MRA), including Phialocephala fortinii, were the most prevalent in all the five bait plants. Eleven of the 123 isolates, after being inoculated onto axenically reared eggplant seedlings, almost completely suppressed the pathogenic effects of a post-inoculated, virulent strain of Verticillium dahliae. Seven of these 11 isolates had come from the roots of eggplant and included Heteroconium chaetospira, P. fortinii, and unidentified species of Fusarium, Penicillium, Trichoderma and MRA. P. fortinii, H. chaetospira, a non-sporulating isolate with white mycelium (SWM) and MRA were easily reisolated from root segments. Hyphae of H. chaetospira, P. fortinii and SWM colonized the root tissues of eggplant without causing apparent pathogenic symptoms. The mechanisms by which these endophytes confer resistance to infection by V. dahliae are unknown but the effectiveness of these fungi in a laboratory setting indicates that they have potential as biocontrol agents and merit further investigation.     

Oral administration of L-citrulline changes the concentrations of plasma hormones and biochemical profile in heat-exposed broilers

We examined the effects of oral administration of L-citrulline (L-Cit) on plasma metabolic hormones and biochemical profile in broilers. Food intake, water intake, and body temperature were also analyzed. After dual oral administration (20 mmol/head/administration) of L-Cit, broilers were exposed to a high ambient temperature (HT; 30 ± 1°C) chamber for 120 min. Oral administration of L-Cit reduced (p < .001) rectal temperature in broilers. Food intake was increased (p < .05) by heat stress, but it was reduced (p < .05) by L-Cit. Plasma levels of 3,5,3′-triiodothyronine, which initially increased (p < .0001) due to heat stress, were reduced (p < .01) by oral administration of L-Cit. Plasma insulin levels were increased by heat exposure (p < .01) and oral L-Cit (p < .05). Heat stress caused a decline (p < .05) in plasma thyroxine. Plasma lactic acid (p < .05) and non-esterified fatty acids (p < .01) were increased in L-Cit-treated heat-exposed broilers. In conclusion, our results suggest that oral L-Cit can modulate plasma concentrations of major metabolic hormones and reduces food intake in broilers.     

Suppression of lipopolysaccharide and galactosamine-induced hepatic inflammation by red grape pomace

Grape pomace is generated in the production process of wine and grape juices and is an industrial waste. This study investigated whether an intake of grape pomace was able to suppress chronic inflammation induced by lipopolysaccharide (LPS) and galactosamine (GalN) in vivo. When Sprague-Dawley rats were orally given methanolic extracts from red and white grape pomace, the extracts inhibited the LPS/GalN-evoked activation of nuclear factor-κB (NF-κB) dose-dependently, and red grape pomace exerted a stronger effect than white grape one. Next, rats were fed an AIN93 M-based diet containing 5% red grape pomace for 7 days, followed by the intraperitoneal injection of LPS and GalN. The intake of the red grape pomace-supplemented diet was found to suppress the LPS/GalN-induced activation of NF-κB and expression of inducible nitric oxide synthase and cyclooxygenase-2 proteins. These results suggest that red grape pomace may contain an abundance of effective compound(s) for anti-inflammatory action.     

Effects of contact with wood on blood pressure and subjective evaluation

This study examined the effects of contact with wood on the living human body using a physiological index and subjective evaluation. Consecutive blood pressure measurements were used as the physiological index, and sensory evaluation using the semantic differential (SD) method was used for subjective evaluation. Consideration was also given to cases in which materials were cooled and heated as well as kept at room temperature, to eliminate the effects of heat flux due to differences in thermal conductivity between wood and other materials. It was found that contact with wood produced coarse/natural sensations, with no associated increase in systolic blood pressure. Contact with cold wood created subjectively dangerous/uncomfortable but still coarse/natural sensations, also with no associated increase in blood pressure; therefore, there was no correspondence between subjective evaluation and physiological responses. Contact with aluminum kept at room temperature and cold acrylic plastic created flat/artificial and dangerous/uncomfortable sensations, with an associated significant increase in blood pressure; thus, there was a close correlation between subjective evaluation and physiological responses. It was therefore concluded that contact with wood, unlike artificial materials such as aluminum, induces no physiological stress even when kept at room temperature or cooled. Part of this report was presented at the 48th Annual Meeting of the Japan Wood Research Society in Shizuoka, April 1998     

Dual effects of reversed winter–spring temperatures on year‐to‐year variation in the recruitment of chub mackerel (Scomber japonicus)

To clarify the effects of temperature on the recruitment of chub mackerel (Scomber japonicus) in the North Pacific, we investigated the influence of winter surface temperature (WST) on spawners at the time of maturity around the spawning grounds and the influence of ambient spring temperature on larvae using estimated temperature (ET) obtained from particle tracking experiments. We found a significant positive correlation between ET approximately 10 days following hatching and the recruitment per spawning stock biomass (RPS) after 2000. The closer (more meandering) the Kuroshio Current (KC) axis was in relation to the spawning ground, the higher (lower) the spring surface temperature and the higher (lower) RPS was in the spawning ground. In contrast, WST inside KC near the maturity/spawning ground was significantly negatively correlated with RPS. A significant negative correlation between the temperatures in winter and spring was detected in the area after 2000, when the conditions of the Pacific decadal oscillation index and the stability of the Kuroshio Extension were synchronous, indicating that KC shifted northward during this time. The reversed temperature pattern was consistent with the winter–spring movement of KC axis in the offshore direction and was correlated with the winter–spring difference in the intensity of the Aleutian low. These results suggest that the annual variation in chub mackerel recruitment after 2000 was strongly affected by the combined effects of ambient temperature because of the reversal of conditions that occurred between winter and spring around the maturity/spawning ground, which was related to the KC path.     

Census and effective population sizes of white‐spotted charr (Salvelinus leucomaenis) in a fragmented landscape

Several habitat models have been proposed to predict population size for stream fishes and to guide habitat assessment and monitoring techniques. However, most models do not incorporate the potential advantage of molecular genetic markers. We conducted a field survey and microsatellite DNA analyses to quantify the relationships among genetic diversity, census/effective population size and habitat variables in fragmented populations of white‐spotted charr (Salvelinus leucomaenis). The census population size significantly increased with the stream length, the number of pools and a pool‐riffle sequence index, a proxy for channel‐unit habitat type complexity within reaches. Population density was correlated with the pool‐riffle sequence index only. Genetic diversity and effective population size were not correlated with the habitat variables or census population size. There was a lack of isolation‐by‐distance population structure in the studied populations. Our results suggest that stream length and the number of pools within reaches associated with habitat complexity are the habitat variables that explain the majority of variation in population size of white‐spotted charr. Our findings provide further evidence that census population size per se is a poor indicator of the inclusive genetic diversity within populations in a fragmented landscape.