Effect of methotrexate exposure at late gestation on development of telencephalon in rat fetal brain

Pregnant rats were treated with 30 mg/kg of methotrexate (MTX) on gestation day (GD) 16, and fetal brains were examined time-dependently. On GD 20, the appearance of the telencephalon in the MTX group was different from that in the control group, and the major axis of the telencephalon of the MTX group was shortened, compared to that of the control group. In the sagittal section of the telencephalon in the MTX group on GD 20, histopathological findings of deformation and narrowing of the cerebral ventricle, the disturbance of the arrangement of the marginal cell layer of subventricular zone (SVZ) and thickening of telencephalic wall, cortical plate and ventricular zone (VZ)/SVZ were possibly attributable to neuronal migration disorders by MTX. Through all the experimental period, few pyknotic cells or TUNEL-positive cells were observed in the VZ/SVZ of the telencephalic wall and striatum in the control group. On the other hand, in the VZ/SVZ of the telencephalic wall and striatum in the MTX group, pyknotic cells or TUNEL-positive cells were observed on GD 17, and they increased significantly on GD18 and then decreased to the control levels from GD 19 onward. The phospho-Histone H3-positive rate decreased remarkedly in the VZ/SVZ of the telencephalic wall and striatum of the MTX group on GDs 17 and 18, compared to the control group, but they recovered on and after GD 19. These results suggested that there was a high possibility that development of the telencephalon in this period required strong folic acid.     

Effects of estrogen on estrogen receptor expression in the bursal cells of chick embryos and steroidogenic enzymes gene expression in the bursa: Relevance of estrogen receptor and estrogen synthesis in the bursa of chick embryos

Estrogen has been reported to act on B cell genesis in the bursa of Fabricius of chick embryos. In this study, we attempted to demonstrate the hypothesis that B cell genesis is controlled by estrogen receptor (ER) in the bursal cells and steroidogenic enzymes synthesized in the bursa. We previously reported the presence of estrogen receptor α (ERα) in the bursa during the late stage of embryogenesis and an increase in the expression of ERα messenger RNA (mRNA) between the 13th day and 16th day. The number of ER-positive cells was maximal on the 16th day. In the present study, ER-positive cells in the bursa during the late stage of embryogenesis increased 4 h after β-estradiol treatment on the 14th to 18th day. The concentration of β-estradiol in the embryonic bursa increased. These results suggest that this stage of embryogenesis is critical in B cell development in the bursa in connection with the effect of estrogen treatment. Our findings also showed that the mRNA expression of five steroidogenic enzymes occurred in the bursa of chick embryos. These results suggest that estrogen is synthesized in the embryonic bursa and estrogen acts on the bursal cells in a paracrine fashion.     

The effect of bark decortication for hiwada production on xylem and phloem formation in Chamaecyparis obtusa

Of all plant materials used to cover the roofs of traditional Japanese buildings, Japanese cypress (Chamaecyparis obtusa) bark, hiwada, has the longest service life and has been used from ancient times. However, wood and bark properties after hiwada harvest have not been evaluated in detail. We studied whether decortication for hiwada production in winter affected xylem and phloem formation. Decorticated trees still preserved all inner bark and part of the outer bark, and both decorticated and control trees had similar annual ring structures at all stem heights in the xylem and phloem. In both xylem and inner bark, no significant difference in ring width at any stem height was found between annual rings before and after decortication. Thus, this study revealed that the decortication of bark for hiwada production does not affect the formation of xylem and the inner and outer bark if decortication is carried out by highly skilled workers in winter.     

转型中的亚洲林业(续二)

在过去的20年里,亚洲林业经历了引人注目的转型,包括从计划经济到市场经济的转变,从天然林供给木材到人工林供给木材的转变,木材替代品与非木质林产品的增加,地方分权与当地群众的参与,生态时代的发展与生态系统管理。在促进亚洲可持续森林资源管理中,市场工具、社区参与和环境服务都得到了应用。亚洲林业的转型表现出多样化,其森林、社会与环境之间的联系是丰富多彩的。     

Experimental studies on rain splash erosion of forest soils after clearing in Okinawa using an artificial rainfall apparatus

The main island of Okinawa, in southwestern Japan, is characterized by a subtropical climate, coral reefs, and reddish soils, which contain Acrisols and Cambisols (FAO/UNESCO classifications). Recently, due to soil erosion on hilly lands, the coral reefs have been damaged by an inflow of reddish soil. Forest clearing is thought to one of the factors increasing rain splash erosion rates on hilly lands, because it is believed that clearing disturbs the forest floor and causes the supply of litter to stop. After clearing, A0, A and B horizons will be denuded, one after the other, by rain splash. In this study, we measured rain splash erosion rates of undisturbed samples of A0, A and B horizons using an artificial rainfall apparatus. The results of experiments on rain splash erosion clarified several aspects of the erosion process in a clearing site. First, it was found that the rain splash erosion rates were higher in fresh litter than in decomposed litter, in decomposed litter than in the A horizon, and in the A horizon than in the B horizon. Thus, the erosion rate increases with soil depth. Secondly, surface-gleyed red and yellow soils are mostly vulnerable to rain splash erosion. The erosion rates of these soils are two to three times higher than those of the red soil and the yellow soils. Thirdly, the erosion rates are affected mainly by bulk density and saturated hydraulic conductivity, not by clay ratio and dispersion ratio. Lastly, the difference between erosion rates of surface-gleyed red and yellow soils and other soils will be even greater if a soil crust has formed, because the formation of a soil crust increases the rain splash erosion rate.     

Potassium distribution in black heartwood of sugi (Cryptomeria japonica) I: Localization in axial parenchyma cells

This is the first report to ascertain potassium location in black heartwood of sugi (Cryptomeria japonica). The objective of this study was to understand the distribution of potassium in sugi black heartwood in connection with anatomical morphology. Scanning electron microscopy with energy dispersive X-ray analysis (SEM-EDXA) was used to investigate the distribution of inorganic elements, especially potassium. In black heartwood, potassium was detected in all tissues (tracheids, ray parenchyma, and axial parenchyma), but its concentrations were different in each xylem tissue. Potassium was particularly accumulated in the droplets in axial parenchyma cells. In addition, calcium was also detected in all tissues. Phosphorus sometimes existed in the granules in ray parenchyma cells. From optical microscopy, the axial parenchyma cells were observed to be arranged in a tangential direction on transverse section and the droplets in these cells were distributed in areas of high ash and green moisture content. For red heartwood, potassium detected by SEM-EDXA was far lower in all tissues than in the black heartwood, while the trend for calcium was the converse. Potassium in axial parenchyma cells was more abundant in black heartwood than in typical pale red heartwood.     

Temporal water deficit and wood formation in Cryptomeria japonica

Cell behavior in the cambium and developing xylem of 3-year-old Japanese cedar (Cryptomeria japonica D. Don.) trees, during and after an 11-day suspension of irrigation, was analyzed. Leaf xylem pressure potential and tangential strain of the stem surface were monitored throughout the experiment. Anatomical features and numbers of developing tracheids and cambial cells were observed in four trees, sampled on Days 0, 4, 8 and 11 after irrigation was suspended. Daytime xylem pressure potential decreased to -1.9 MPa on Day 7 and remained the same until irrigation was resumed on Day 11. The transverse dimensions of the tracheids, which began to form secondary walls, began to decrease on Day 4. The number of cells in the cambial zone and cell expansion zone decreased abruptly on Day 8. Tangentially aligned developing tracheids with collapsed cell walls were observed in samples harvested on Days 8 and 11. Secondary wall formation was recognized in these tracheids. After the resumption of irrigation, xylem pressure potential recovered rapidly to the same value as before the suspension of irrigation. Tangential strain increased within 30 min after the resumption of irrigation, and continued to increase until the onset of light the next day. Eighteen days after the resumption of irrigation, anatomical features of cells in the cambium and cell-expansion zone were similar to those observed before suspension of irrigation.     

Cellular localization of IL-18 and IL-18 receptor in pig anterior pituitary gland

Pro-inflammatory cytokine interleukin 18 (IL-18) has been proposed to have a role in modulating immuno-endocrine functions. Our previous study showed that IL-18 and IL-18 receptor (IL-18R) colocalized in somatotrophs of the bovine anterior pituitary gland, and the possibility that IL-18 acts on somatotrophs as an autocrine factor. In the present study, we investigated the localization of IL-18 and IL-18R in the pig anterior pituitary gland. RT-PCR analysis showed the expression of IL-18 and IL-18R mRNAin the pig anterior pituitary gland. Immunohistochemistry of IL-18 and specific hormones revealed the presence of IL-18 in somatotrophs, mammotrophs, thyrotrophs and gonadotrophs. IL-18R was localized in somatotrophs and thyrotrophs. Furthermore, the somatotrophs immunoreactive for IL-18 did not contain IL-18R. Thus, IL-18R and IL-18 were not colocalized in an identical somatotroph. These findings suggest that the localization of IL-18 in pig somatotrophs is different from that in bovine somatotrophs, although IL-18 closely associates with somatotrophs in the anterior pituitary glands in both species.     

Prevention of immune responses to human erythropoietin in cynomolgus monkeys (Macaca fascicularis)

Genes and proteins of human origin are often administered to monkeys for research purposes, however, it can be difficult to obtain sufficient levels of the products in vivo due to immunological clearance. In this study, we showed that human erythropoietin (hEPO) induces generation of anti-hEPO antibody in cynomolgus macaques (n=2), although 92% of amino acid residues are common between the human and macaque EPO. The administered hEPO was thus eliminated from the animals. On the other hand, when an immunosuppressant, cyclosporin A (CyA), was administered (6 mg/kg) intramuscularly every other day in combination with hEPO (n=2), no anti-hEPO antibody was generated and high serum levels of hEPO were obtained during administration of hEPO, resulting in an increase in serum hemoglobin levels. No adverse effects associated with CyA were observed. Thus, CyA treatment is useful for prevention of immune responses associated with the administration of human proteins in monkeys.     

Characterization of Bovine herpesvirus type 4 isolated from cattle with mastitis and subclinical infection by the virus among cattle

An outbreak of contagious mastitis occurred among cattle on a farm, and bovine herpesviruses were isolated from the affected mammary tissues, scabs and abscess discharge of the cattle. A bovine herpesvirus type 4 (BoHV-4)-specific fragment was amplified from the isolates by polymerase chain reaction (PCR). Restriction endonuclease analyses demonstrated that the isolates were related to Movar-like European type BoHV-4. To determine the ratio of BoHV-4 subclinical infection in the cattle, a genomic survey was performed by PCR for cattle that were moved to the animal hygiene service station in Ibaraki prefecture. The BoHV-4 genome was occasionally detected in peripheral blood leukocytes, lymph nodes and nervous tissues. The rate of BoHV-4 subclinical infection was relatively high in the cattle.