Inflammation of the cardiac coronary artery in ICR mice

Inflammation of the cardiac coronary artery in ICR mice is occasionally observed in toxicity studies; however, this has not been well explored histologically. Herein, we investigated the detailed histology of the associated lesions in 6–8-week-old ICR mice. Coronary artery inflammation in the right ventricular wall was observed in 10 of 142 mice (7.0%). Histopathological examination revealed hypertrophy of the vascular smooth muscle cells and perivascular infiltration of macrophages in mild cases. In moderate to marked cases, single-cell necrosis of vascular smooth muscle cells, hemorrhage of the tunica media, and fibrinoid necrosis of the vessel wall were observed, in addition to the changes seen in mild cases. Electron microscopic examination of moderate cases revealed a discontinuous internal elastic lamina suggestive of rupture, and vascular smooth muscle cells beneath the elastic lamina showed degeneration and necrosis. These findings suggest that the lesions developed as a rupture of the internal elastic lamina and necrosis of vascular smooth muscle cells, while leaked plasma components caused vascular and perivascular inflammation. In ICR mice, dystrophic calcinosis (DCC) is known to occur rarely in the right ventricle. DCC is defined as focal calcification in necrotic myocardial fibers, the pathogenesis of which is considered to involve ectopic calcification. Since calcification was not observed in any part of the heart, including the inflammation region, the pathophysiology of cardiac arterial inflammation seen in our ICR mice was considered to differ from that of DCC.     

Pathological study of lead poisoning in whooper swans (Cygnus cygnus) in Japan.

During spring 1989, thirty-three whooper swans (Cygnus cygnus) died at Lake Miyajima in Hokkaido, Japan; 15 were examined. The birds were diagnosed as having subacute lead poisoning due to ingestion of spent lead shot. The main gross findings were bile-stained liver, edematous or gelatinous bone marrow, bile-stained lining with hyperkeratosis and lead pellets in the gizzard, and proventricular impaction. Histopathologically, there was lead-hemolytic jaundice of the liver, hemosiderosis in the liver and spleen, and hypoplasia of the bone marrow with increased numbers of polychromatic erythroblasts. Acid-fast intranuclear inclusion bodies were seen in kidneys of seven swans. Under electron microscopy, inclusion bodies had frayed contours and consisted of high-electron-dense fine granules. The lead concentration of the liver ranged from 5.5 to 44.3 mg/kg wet weight. It was suggested that these changes resulted from excess breakdown of erythrocytes, inhibition of heme synthesis, and impaired erythropoiesis caused by lead shot.     

The cumulative methane production from dairy cattle slurry can be explained by its volatile solid,temperature and length of storage

In order to refine the national estimate of methane emission from stored cattle slurry, it is important to comprehend the basic characteristics of methane production. Two dairy cattle slurries were obtained from livestock farms located in Hokkaido (a northern island) and Kyushu (a southern island). The slurries were diluted with water into three levels: undiluted, three times diluted, and 10 times diluted. Three hundred mL of the slurries were put into a bottle with a headspace volume of 2.0 L, which was filled with nitrogen gas and then sealed by butyl rubber. Four levels of temperature were used for incubation: 35, 25, 15 and 5 °C. The time course of the cumulative methane production per volatile solid (VS) was satisfactorily expressed by an asymptotic regression model. The effect of dilution on the methane production per VS was not distinctive, but that of temperature was of primary importance. In particular, higher temperature yields a higher potential production and a shorter time when the cumulative production reaches half of the potential production. The inclusive and simple models obtained in this study indicate that the cumulative methane production from stored cattle slurry can be explained by VS, temperature and length of storage.     

Inhibiting factors of symptom development in several Japanese red pine (<Emphasis Type="Italic">Pinus densiflora</Emphasis>) families selected as resistant to pine wilt

Pinus densiflora and P. thunbergii, native to Japan, are highly susceptible to pine wilt disease caused by infection with a pine wood nematode (Bursaphelenchus xylophilus). Trees of these susceptible species have occasionally been found surviving in forests that are extensively damaged by this disease. Seedlings from a part of surviving trees that were selected as resistant families indicate lower mortality rates after the infection. The factors that prevent the symptoms from developing in resistant families of a susceptible species, P. densiflora, as based on the analysis of the pathogens behavior in the tree tissue and the anatomy of the resistant families, are presented in this paper. Nematode populations remained lower in the stems of seedlings from resistant families of P. densiflora than in the stems of non-resistant families. Areas dysfunctional in water transport developed in the stems of resistant families, but did not reach a size large enough to seriously block the ascent of sap. These results suggest that there are systems within the seedlings that prevent nematode migration and reproduction. The 2-year-old seedlings from resistant families of P. densiflora, however, did not suppress the pathogen activity. Numerous branches are a visible characteristic in the seedlings of some resistant families. The arrangement of the resin canals, the only channels in the tree to the pathogen migration, was disoriented at the joints between the branches and the main stem. Such a structure may be effective as a barrier to nematode dispersal.     

Effects of impregnation of simple phenolic and natural polycyclic compounds on physical properties of wood

The impregnation of various simple phenolic and natural polycyclic compounds into wood was investigated from the viewpoints of vibrational property and dimensional stabilizing effect. When simple phenolic compounds were impregnated, the loss tangent (tan ) in the longitudinal direction increased linearly with increasing weight gain. Meanwhile, among the natural polycyclic compounds hematoxylin decreased the tan drastically by impregnation. It was suggested that the five hydroxyl groups and the pyran ring oxygen in the hematoxylin molecule contribute to formation of the crosslinkage-type hydrogen bonds between wood components. The rigidity of hematoxylin molecules may also be important. By impregnation of about 10% catechol, resorcinol, and saligenin, a 40% level of antiswelling efficiency (ASE) was attained, although a significant dimensional stabilizing effect was not observed after impregnation of natural polycyclic compounds.Part of this work has been published as a Rapid Communication inMokuzai Gakkaishi 43(12). It was also presented at the 48th annual meeting of the Japan Wood Research Society, Shizuoka, April 1998     

Identification of marker genes for intestinal immunomodulating effect of a fructooligosaccharide by DNA microarray analysis

Prebiotic fructooligosaccharides are noted for their intestinal immunodulating effects, and the identification of markers for the effects is a matter of great concern. This study aimed to identify marker genes for physiological effects of a particular fructooligosaccharide (FOS) on a host animal and also to define the target of its function in the small intestine. DNA microarray technology was used to screen candidate marker genes, and comprehensive changes in gene expressions in the ileum of mice fed with FOS were investigated. One of the major physiological effects of FOS was intestinal immunomodulation. Marker genes were then identified for major histocompatibility complex classes I and II, interferon, and phosphatidylinositol metabolites. Also, the ileum was segmented into Peyer's patch (PP) and the other ileal organ (DeltaPP), and these were analyzed by quantitative RT-PCR method, with the result that the site for recognizing the FOS function was the DeltaPP rather than the PP. This is the first paper showing the markers for the physiological effects of FOS in the small intestine at gene expression level. Applying these marker genes would make it possible to clarify the mechanisms of how the administration of dietary FOS and associated changes in the intestinal environment are recognized by host organisms as well as how its immunomodulating effects are expressed in the body.     

Occurrence of orally administered mulberry 1-deoxynojirimycin in rat plasma

1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, is a characteristic constituent of the mulberry leaf. Dietary mulberry DNJ may be beneficial for the suppression of abnormally high blood glucose levels, thereby preventing diabetes mellitus. Although there is considerable interest in the effects of mulberry DNJ, the intestinal absorption and pharmacokinetic profile of orally administered mulberry DNJ have never been characterized. In this study, we developed a method for determining the level of plasma DNJ by hydrophilic interaction chromatography coupled to a mass spectrometric detector (HILIC-MS) to investigate the absorption and metabolism of orally administered mulberry DNJ in rats. DNJ was separated from plasma extract on a TSK gel Amide-80 column, a representative column for HILIC. At postcolumn, DNJ was concurrently detected and identified by MS. The plasma DNJ concentration in fasted rats was below the detection limit [<1 microg (6 nmol)/mL]; however, the concentration reached a maximum [15 microg (92 nmol)/mL] 30 min after the administration of mulberry DNJ (110 mg/kg of body weight), and the DNJ concentration decreased rapidly thereafter. When the rats received different amounts of mulberry DNJ (1.1, 11, and 110 mg/kg of body weight), dose-dependent incorporation of DNJ into the plasma was confirmed. We did not detect any DNJ metabolites in the plasma. These findings indicate that orally administered mulberry DNJ is absorbed as an intact form from the alimentary tract and then is quickly excreted from the body. The developed HILIC-MS method could be applied in determining levels of DNJ in urine and tissues, and therefore, the method would be a powerful tool for studying the metabolic fate of mulberry DNJ as well as its bioavailability.     

Isolation and characterization of eight microsatellite loci in the Otton frog, Babina subaspera

The Otton frog, Babina subaspera, is an endangered forest frog species with unusual morphological and ecological features compared with other frogs. Eight microsatellite markers were isolated from B. subaspera and characterized with 24 individuals from Amami Islands, Kagoshima prefecture, Japan. The number of alleles observed for each locus ranged from 9 to 19 (average 13.9). The observed heterozygosity (H _O) and the expected heterozygosity (H _E) ranged from 0.71 to 0.92 and 0.83 to 0.92, respectively. These microsatellites will be useful for future genetic studies of B. subaspera, which will be essential in the conservation of this species.     

Identification of small molecule activators of cryptochrome

Impairment of the circadian clock has been associated with numerous disorders, including metabolic disease. Although small molecules that modulate clock function might offer therapeutic approaches to such diseases, only a few compounds have been identified that selectively target core clock proteins. From an unbiased cell-based circadian phenotypic screen, we identified KL001, a small molecule that specifically interacts with cryptochrome (CRY). KL001 prevented ubiquitin-dependent degradation of CRY, resulting in lengthening of the circadian period. In combination with mathematical modeling, our studies using KL001 revealed that CRY1 and CRY2 share a similar functional role in the period regulation. Furthermore, KL001-mediated CRY stabilization inhibited glucagon-induced gluconeogenesis in primary hepatocytes. KL001 thus provides a tool to study the regulation of CRY-dependent physiology and aid development of clock-based therapeutics of diabetes.     

Toxicity of Nicotine by Repeated Intratracheal Instillation to F344 Rats

In vivo, nicotine in cigarette smoke induces various effects not only on the respiratory system but also the central and peripheral nerve systems, circulatory organs and digestive organs, and there is a possibility of promotion of lung tumorigenesis. The present experiment was conducted to examine histopathological changes caused by nicotine in the lung with repeated intratracheal instillation (i.t.). Six-week-old male F344 rats were administered nicotine by i.t. at doses of 0.05, 0.1 and 0.2 mg nicotine/rat every 3 weeks beginning at week 4, for up to a total of 9 times and were then sacrificed at week 30. The total number of administrations, total dose of nicotine and effective number of rats were 9 times, 0.45 mg and 5 rats and 4 times, 0.20 mg and 5 rats for the 0.05 mg nicotine/rat group; 3 times, 0.30 mg and 5 rats and 4 times, 0.40 mg and 3 rats for the 0.1 mg group; and 3 times, 0.60 mg and 3 rats for the 0.2 mg group, respectively. As a control group, 5 rats were administered 0.2 ml saline/rat 9 times. Some rats administered 0.1 and 0.2 mg nicotine suffered convulsions just after administration. Histopathologically, though proliferative changes were not observed, neutrophil infiltration, edema and fibrosis in the lung were induced by nicotine. In conclusion, repeated treatment of nicotine promoted neurologic symptoms in the acute phase, and strong inflammation in the lungs in the chronic phase, even at a low dose. Toxicity of nicotine is suggested to depend not on total dose of nicotine in the experiment but rather on repeated injury with consecutive administration.