Influence of stem rot pathogen <Emphasis Type="Italic">Fomitiporia</Emphasis> sp. on “Sanbu-sugi” cultivar of the Japanese cedar <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

An unidentified Fomitiporia sp. causes severe white-rot on stems of a cultivar “Sanbu-sugi” of the Japanese cedar, Cryptomeria japonica. The influence of the fungus on tree health and wood properties was determined. Bark from dead branches and xylem from living branches contained more glucose than bark from living branches and xylem from dead branches. Tree heights at which annual rings were disconnected were 2, 4 and 6 m at ages 37, 15 and 24 years old, respectively. The pH values of damaged parts were lower than those of non-damaged parts, and the damaged parts were clearly identified using bromocresol green solution. Weight loss of sapwood during 60 days of fungal degradation was 1.4 %, which was less than that by a saprophyte, Trametes versicolor. The amount of polyphenols in the heartwood from damaged tree stems was less than that from non-damaged stems. Degraded parts were less stiff than the non-degraded sapwood as measured with a wood-decay tester, Pilodyn. Our observations indicate that damaged stems are chemically and physically inferior to non-damaged stems.     

Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)

We succeeded to establish cell lines from endemic fish species Honmoroko Gnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary of G. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function.     

Porcine MHC classical class I genes are coordinately expressed in superantigen-activated mononuclear cells

The expression of the major histocompatibility complex (MHC) classical class I genes is important for the adaptive immune response to target virus-infected cells and cancer cells. The up-regulation of the MHC is achieved by hormonal/cytokine signals including IFN-γ-inducible elements. The swine leukocyte antigen (SLA), the MHC class I region of pigs, consists of the duplicated classical class I genes, SLA-1, SLA-2 and SLA-3, but the molecular mechanisms involved in their up-regulation after T cell stimulation have not been fully elucidated. In order to better understand some of the putative regulatory mechanisms of SLA class I gene expression in activated T cells, we examined the coordinated expression of the SLA classical class I, IFN-γ and interferon regulatory factor-1 (IRF-1) genes in the peripheral blood mononuclear cells (PBMCs) of SLA homozygous Clawn miniature swine stimulated for 72h with either IFN-γ or an enterotoxin produced by Staphylococcus aureus. This enterotoxin, toxic shock syndrome-1 (TSST-1), is known to act as a superantigen (sAG) to activate the T cells in various vertebrate species. We showed by using mAbs and flow cytometry that the CD4(+)CD25(+) cell number of swine PBMCs was also increased by TSST-1 and to a lesser degree by IFN-γ. Time course analyses of the expression of the IFN-γ, IRF-1 and the three classical class I genes, SLA-1, SLA-2, and SLA-3, in PBMCs by quantitative real-time PCR revealed a transitory response to TSST-1 or IFN-γ stimulation. The IFN-γ mRNA levels in the PBMCs were continuously up-regulated over the first 48h by TSST-1 or IFN-γ. In contrast, SLA class I expression moderately increased at 24h and then decreased to a baseline level or less at 72h of IFN-γ or TSST-1 stimulation. The three classical SLA class I genes showed similar expression kinetics, although SLA-3 mRNA level was consistently lower than those of SLA-1 and -2. The expression of IRF-1, a modulator of SLA expression, showed similar kinetics to those of the three classical SLA class I genes. The expression profiles detected by flow cytometry of the SLA molecules on the cell surface of PBMCs were maintained at a consistently high level during cell stimulation with either TSST-1 or IFN-γ, which was distinct from the kinetics of mRNA expression. These results showed that miniature swine SLA class I mRNA expression was effectively and equally up-regulated among the three loci and coordinately with IRF-1 gene expression after stimulation of T cell activation by sAG or IFN-γ.     

Two Types of Grapevine Yellow Speckle Viroid 1 Isolated from Commercial Grapevine Had the Nucleotide Sequence of Yellow Speckle Symptom-inducing Type

Grapevine yellow speckle viroid 1 (GYSVd 1) was first detected in Japan from commercial grapevines. The viroid seems to be spreading in Japanese commercial vineyards but without causing noticeable disease symptoms. Two Japanese GYSVd 1 isolates from the cultivars Campbell and Steuben were sequenced. The Campbell isolate is included in the type 2 variant of GYSVd 1, and the Steuben isolate is included in the type 3. Both of the isolates were grouped into the yellow speckle-inducing variants in the nucleotide sequence ; however, the etiological importance of this viroid on the viticulture of Japan has not yet been determined. Received 14 June 1999/ Accepted in revised form 25 August 1999     

Lignin-degrading activity of edible mushroom <Emphasis Type="Italic">Strobilurus ohshimae</Emphasis> that forms fruiting bodies on buried sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) twigs

Strobilurus ohshimae is an edible mushroom, and it specifically forms its fruiting bodies on buried sugi (Cryptomeria japonica) twigs. In this research, we studied lignindegrading activity of S. ohshimae. We isolated 18 strains of S. ohshimae from various regions of Japan, and determined their lignin degradation rates on sugi wood meal medium. All the strains of S. ohshimae degraded approximately 6%–12% of sugi lignin in 30 days, and these lignin degradation rates were 1.5–3 times higher than those of Trametes versicolor, which is a typical lignin-degrading fungus. Among the three main lignin-degrading enzymes, activity of lignin peroxidase and manganese peroxidase was not observed, while 4340U/g of laccase was produced in 30 days. To investigate the effect of wood species on lignin degradation by S. ohshimae, the lignin degradation rate and laccase productivity on sugi wood meal medium were compared with those on beech (Fagus crenata). In T. versicolor, both lignin degradation rate and laccase productivity were higher on beech than on sugi. Conversely, in S. ohshimae, lignin degradation rate and laccase productivity were higher on sugi than on beech. Therefore, it was suggested that coniferous lignin is not always difficult to degrade for the fungi that inhabit softwood. Part of this article presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

Relationship between blood calcium level and ST peak interval of electrocardiographic variables in peripartum Holstein cows

The association between blood calcium levels and electrocardiographic variables was compared in 137 normal parturient and 36 peripartum recumbent Holstein cows to determine whether hypocalcemia in peripartum dairy cows can be rapidly diagnosed using electrocardiograph. Inverse of STc (ST peak interval/SS interval^0.5) and blood ionized calcium or serum calcium concentrations were strongly correlated, and both correlation coefficients were 0.81 (P<0.001). The 95% prediction interval indicated that cows with STc >0.385 ± 0.001 sec are very likely to be hypocalcemic (blood ionized or serum calcium concentrations of <0.9 mmol/l or <7.5 mg/dl, respectively). These findings indicate that hypocalcemia in parturient cows can be non-invasively estimated using the STc.     

Sardine and anchovy regime fluctuations of abundance in four regions of the world oceans: a workshop report

Regimes of high abundance of sardine (Sardinops sagax and Sardina pilchardus) have alternated with regimes of high abundance of anchovy (Engraulis spp.) in each of the five regions of the world where these taxa co-occur and have been extensively fished. When one taxon has been plentiful, the other has usually been at a reduced level of abundance, and vice versa. Changes in the four heavily fished regions that support S. sagax–the Japanese, Californian, Humboldt, and Benguela systems–from a regime dominated by one taxon to a high level of abundance of the other have occurred more or less simultaneously. In the Pacific Ocean, sardines have tended to increase during periods of increasing global air and sea temperatures and anchovies to decrease. The Japanese system is dominated by sardines to a greater extent than the other systems, and sardines off Japan appear to increase as the Kuroshio Current cools. At the eastern edge of the Pacific Ocean, sardines colonize cooler areas during periods of warming. The Benguela system is out of phase with the three Pacific systems. The four systems all appeared to be in a state of flux in the 1980s. Increased abundance of the subdominant taxon is often one of the first signs of change. Sardines are relatively sedentary in refuge areas when scarce but change behavior to become highly migratory and colonize cooler areas when abundant. Anchovies, by contrast, expand around a fixed geographic center.     

Possible mechanisms underlying fluctuations in the Far Eastern sardine population inferred from time series of two biological traits

A cycle of fluctuation in the Far Eastern sardine, Sardinops sagax, population is accompanied by an abrupt drop and recovery of fish size and lipid content of parent fish, which may reflect a phase variation from coastal to oceanic and its reverse. The cycle begins with the survival success of a smaller number of eggs produced by high-quality parents, which is triggered by a global climate change (density-independent process), and finally ends in the survival failure resulting from an overexpansion of an enormous number of fish produced by low-quality parents (density-dependent process).     

Effect of fatty acids added to the milk replacer on white scour and excretion of fatty acids in Holstein calves

In order to examine the relationship between white scour and fatty acids, we added fatty acids to the milk replacer. Twenty healthy Holstein calves were divided into 4 groups, five calves per group; a control group with no fortified fatty acid, and 3 groups fed either with oleic, stearic, or palmitic acid, respectively. The calves were fed milk replacer (5% of the calf's body weight) twice a day but the fatty acids (2 wt % of milk replacer) were added only once. The fecal and blood samples were obtained at 0, 12, 24, 36, and 48 h after feeding of the acids. All five calves in the palmitic acid group, and 3 out of 5 calves each in the stealic and the oleic acid groups had whitish feces after feeding fatty acid. The stearic acid group had a significantly elevated stearic acid concentration in the feces during 24–36 h compared to the pre-feeding level. The fecal concentration of palmitic acid increased significantly at 24–36 h in the palmitic acid group. We concluded that the long-chain saturated fatty acids are one of the causes of white scour in calves.