Biosafety assessment of transgenic poplars overexpressing xyloglucanase (AaXEG2) prior to field trials

We performed biosafety assessments of transgenic poplars prior to field trials. Constitutive expression of the Aspergillus aculeatus xyloglucanase in Populus alba increased the cellulose content and specific gravity of its stem, the leaves of which were visibly greener, thicker, and smaller than those of the wild-type plant. Although the young transgenic poplars grew faster than the wild type in a growth chamber, there was no distinguishable difference in growth between the poplars when they were placed in a special screened greenhouse. Allelopathic tests showed that the transgenic poplars do not produce harmful substances. Based on all the biosafety assessments and the scientific literature on poplar species, we came to the conclusion that transgenic poplars probably do not disturb the biological diversity of the surrounding environment, even when they are submitted to field trials.     

Change in river water chemistry as affected by the confluence of forest drainage water in dairy farming area on Hokkaido Island, Northern Japan

We evaluated i) the difference in river water chemistry between a watershed mainly consisting of pasture and a watershed mainly consisting of forest, and ii) how the chemistry of river draining the pasture is influenced by that of the river draining the forest. We selected one river (designated as the T-river) draining the pasture (3,587 ha), and two rivers draining a forest (738 and 879 ha) in eastern Hokkaido, northern Japan. During higher river flow due to precipitation and thawing, the concentrations of NO ₃ ⁻ , SO ₄ ²⁻ , K⁺, Fe, and Al increased, suggesting the relative importance of the shallow soil layer as their source. On the other hand, Na⁺, Mg²⁺, Ca²⁺, and Si decreased, suggesting the relative importance of the source in a deep soil layer. The concentrations of NO ₃ ⁻ , Cl⁻, SO ₄ ²⁻ , K⁺, Na⁺, Mg²⁺, Ca²⁺, and Fe were higher in the T-river than in the forest drainage waters, suggesting the contribution of the excretion components from the milk cows. The Si concentration exhibited the opposite pattern. The concentrations of NO ₃ ⁻ , Cl⁻, SO ₄ ²⁻ , K⁺, Na⁺, Mg²⁺, Ca²⁺ (p<0.001), and Fe (p<0.05) in the T-river decreased after the confluence of the forest drainage waters, while Si concentration increased (p<0.001). The reason for the change in river chemistry was the confluence of the forest drainage waters. These findings suggested the environmental role of the forest in the dilution of the polluted river. Prof. S. Ohata, and Prof. H. Takeda, Graduate School of Agriculture, Kyoto University, facilitated this study. Prof. T. Sakai, Graduate School of Informatics, Kyoto University, offered this study some convenience. Dr. M. Sakimoto, and Dr. M. Katsuyama, Graduate School of Agriculture, Kyoto University, offered useful advise.     

Volatile organic compounds from wood and their influences on museum artifact materials II: Inference of causal substances of deterioration based on intercomparison of laser Raman spectra of deteriorated products

Volatile organic compounds (VOCs) from wood and their effects on artifacts are of intense interest to museum staff who must strictly manage the preservation environment. In this study, the causal substances of deterioration were inferred by comparison with laser Raman (LR) spectra of deterioration products. The artifact samples were deteriorated using VOCs from woods (WV test), and using specific substances that occur in wood VOCs (SV test). The deteriorated samples were analyzed by LR spectroscopy and the results of the WV and SV tests were compared. Hinokitiol and acetic acid were found to be two of the main causal substances of deterioration in western red cedar. Deterioration of iron and copper by spruce did not appear to be caused by acetic acid, as is generally assumed, but by some other unknown compound(s). Sensitivity to wood-based VOC components depended on the type of artifact. In western red cedar, mitsudasou (litharge) was very sensitive to acetic acid, while other artifacts were sensitive to hinokitiol. The LR method used in this study is very useful for the inference of causal substances of deterioration based on the detection of small amounts of deteriorated products such as those generated by deterioration due to wood VOCs. Part of this article was presented at the 25th Annual Meeting of the Japan Society for the Conservation of Cultural Property, Kyoto, June 2003     

Pericardial mesothelioma in a neonatal calf

We encountered an extremely rare tumor, a pericardial mesothelioma, in a neonatal calf. The patient calf showed severe abdominal distention, and died immediately after birth. The thoracic cavity was contained a huge heart with a large amount of pericardial fluid. A number of granular and cobblestone-like nodules were dispersed over the epicardium and pericardium. The nodules consisted of papillary proliferations of neoplastic cells, and the neoplasm occasionally showed mesenchymal proliferations. Immunohistochemistry revealed that they had the characteristics of mesothelial cells (cytokeratin-and vimentin-positive), and the neoplasm was diagnosed as mesothelioma.     

Parathyroid hormone-related protein (PTHrP) and Ca levels in the milk of lactating cows

To investigate the roles of mammary PTHrP in calcium uptake and/or release in the mammary gland of cows, plasma PTHrP and Ca levels, and their arterial-venous differences were examined in a Jersey cow during the periparturient period. Levels of Ca in both abdominal aorta and abdominal subcutaneous vein blood slightly decreased around the parturition and at 24 days after the parturition, however, no remarkable arterial-venous differences were observed. Plasma PTHrP levels in both arterial and venous samples were below the detection limit (0.57 pmol/l) during the experimental period. Milk PTHrP and Ca levels were measured in 9 Holstein dairy cows. Although plasma PTHrP levels in all arterial and venous samples were also below the detection limit, milk PTHrP and Ca levels were remarkably high, ranging from 14,900 pmol/l to 41,200 pmol/l and from 772 mg/l to 1,200 mg/l, respectively. In addition, a significant positive correlation (P<0.01) was observed between milk PTHrP and Ca levels. These results suggested that mammary PTHrP is closely related to Ca concentration in the milk.     

Analysis of neutral glycosphingolipids from Trypanosoma brucei

Neutral glycosphingolipids (GSLs) were isolated from Trypanosoma brucei and analyzed by thin-layer chromatography (TLC), TLC/secondary ion mass spectrometry (TLC/SIMS), and liposome immune lysis assay (LILA). Three species of neutral GSLs, designated as N-1, -2, and -3 were separated on TLC. N-1 GSL migrated very close to glucosylceramide (GlcCer) and N-2 GSL showed the same mobility as lactosylceramide (LacCer). On the other hand, the mobility of N-3 GSL on the TLC plate was slower than globotetraosylceramide (Gb4). In order to characterize the molecular species of neutral GSLs from T. brucei, N-1, -2 and -3 GSLs were analyzed by TLC/SIMS. The TLC/SIMS analysis of N-1 of the parasites revealed a series of (M–H)⁻ ions from m/z 698 to 825 representing the molecular mass range of ceramide monohexoside (CMH) (GlcCer or galactosylceramide). On the other hand, the TLC/SIMS spectra of N-2 GSL revealed a series of (M–H)⁻ ions from m/z 944–987 indicating the molecular mass range of LacCer. In the TLC/SIMS analysis of N-3 GSL, however, the characteristic molecular ions that can elucidate the structure of N-3 GSL were not obtained. In order to confirm the results obtained from TLC/SIMS, N-1, -2, and -3, GSLs were tested by LILA with specific antibodies against GlcCer, LacCer, and Gb4, respectively. N-1 GSL had reactivity to anti-GlcCer antibody and N-2 GSL reacted with the antibody against LacCer. However, N-3 GSL was not recognized by anti-Gb4 antibody. Using anti-GlcCer and anti-LacCer antibodies, furthermore, we studied the expression of GlcCer and LacCer in T. brucei parasites. Both GlcCer and LacCer were detected on the cell surface of T. brucei.     

Changes in circulating adiponectin and metabolic hormone concentrations during periparturient and lactation periods in Holstein dairy cows

Although our previous report demonstrated that adiponectin and AdipoR1 gene expressions changed among different lactation stages in the bovine mammary gland, its in vivo kinetics remain unclear in ruminant animals. In this study, we investigated the changes in circulating concentrations of adiponectin, as well as other metabolic hormones and metabolites, (i) during the periparturient period and (ii) among different lactation stages, in Holstein dairy cows. In experiment 1, serum adiponectin concentrations increased after parturition. Serum insulin concentrations were lower in the postpartum than prepartum period, whereas serum growth hormone (GH) concentrations increased in the postpartum period. Serum nonesterified fatty acids (NEFA) levels were increased during the postpartum period and were dependent on the parity. In experiment 2, there was no significant difference in plasma adiponectin concentrations among lactational stages. Plasma insulin concentrations tended to be lower in early lactation while plasma GH levels tended to be higher. Plasma NEFA concentrations were significantly lower in mid‐ and late‐lactation stages than non‐lactation stages. These findings indicate that elevation of serum adiponectin might be involved in energy metabolism just around parturition, and might exert its action through regulation of receptor expression levels in target tissues in each lactational stage in Holstein dairy cows.     

Japanese black cattle with ateliosis showed lower insulin responses during glucose tolerance test

In order to determine insulin secretability and glucose utilization, a glucose tolerance test was performed in ateliotic cattle of 2 paternal strains; MHO and HSK cattle. MHO and HSK cattle showed different endocrine patterns in our previous study. Area under the insulin concentration curves (insulin-AUC) in the ateliotic cattle were significantly lower (122.3 +/- 59.4 ng.min/ml and 99.2 +/- 24.8 ng.min/ml for MHO and HSK cattle, respectively) than the control cattle (420.2 +/- 175.2 ng.min/ml). These low insulin responses to GTT may have an influence on growth retardation in MHO and HSK cattle.     

Detection of fungi producing infection-inhibiting metabolites against <Emphasis Type="Italic">Alternaria alternata</Emphasis> Japanese pear pathotype from fungi inhabiting internal tissues of Japanese pear shoots

Fungi inhabiting Japanese pear were isolated from internal tissues of cv. Nijisseiki, and culture filtrates (CFs) of 100 isolates were evaluated for their inhibitory activity against infection by Alternaria alternata Japanese pear pathotype. CFs of 11 isolates inhibited lesion formation on the pear by the pathogen. Among these isolates, CFs of five isolates inhibited spore germination. CFs of the six other isolates inhibited appressorial formation, infection hypha formation, AK-toxin production, or a combination of these actions. Analysis of sequence homology in the rDNA ITS1 regions of these isolates showed that most isolates had high homology with some fungal endophytes.