Re-evaluation of the colorimetric assay for cytidine deaminase activity

This study re-evaluated the colorimetric assay for cytidine deaminase (CTD), and showed that the optimum conditions were pH 7·5, 37°C, and up to 24 h. In addition, this method was found to require protein precipitation. Following these modifications, intra-assay and inter-assay coefficients of variation were below 5 per cent, indicating that the assay was highly reliable. CTD activity was determined in 282 serum samples from 206 normal pregnant women by the incubation of 100 μl of serum and 400 μl of 1·4 mmol/l cytidine substrate for 16 h at 37°C. Following protein precipitation, the ammonia liberated during conversion was measured by a colorimetric procedure. The mean (±SD) CTD activity was 7·31 ± 2·50 U at 3–12 weeks of gestation, 8·70 ± 2·12 U at 13–24 weeks, 7·59 ± 2·25 U at 25–36 weeks, and 7·29 ± 2·16 U at 37–42 weeks. High levels of CTD activity were found in patients with abruptio placentae and amnionitis associated with intrauterine fetal death (IUFD). The increase in CTD activity was noted from 3 days to 1 week before the confirmation of IUFD. The placenta contains extremely high levels of CTD, but cord serum does not. Thus, the excessive elevation of CTD activity was probably derived from progressive placental damage. This modified CTD assay was concluded to be simple and reliable, and may perhaps be useful in detecting pregnancy disorders.     

Evaluation of metals in the residue of paper sludge after recovery of pulp components using an ionic liquid

The quantity of sludge produced by the paper industry in Japan in 2011 was estimated to be 27.91 million tons wet weight. This amount is the third largest among all industrial wastes. To explore ways of reusing recovered paper pulp and safely disposing of the residue, we investigated the distribution of metals in the process of recovery of pulp from two types of paper sludge using an ionic liquid. 32 and 46 % of pulp from paper sludge A and B was recovered using ionic liquid, respectively; the resultant weight reduction of the solid mass was comparable to that of incineration. There were virtually no heavy metals but aluminum in the recovered pulp, which makes its reuse viable at present. The characteristics of the metals in the residues differed according to the treatment used and properties of the paper sludge. Copper accounted for ~10 % in the ionic liquid used for recovery of pulp from the paper sludges. This suggests that [bmIm]Cl has the potential to dissolve a specific amount of copper compared to other metals.     

Polychlorinated biphenyl (118) activates osteoclasts and induces bone resorption in goldfish

To analyze the effect of polychlorinated biphenyl (PCB) 118 on fish bone metabolism, we examined osteoclastic and osteoblastic activities, as well as plasma calcium levels, in the scales of PCB (118)-injected goldfish. In addition, effect of PCB (118) on osteoclasts and osteoblasts was investigated in vitro. Immature goldfish, in which the endogenous effects of sex steroids are negligible, were used. PCB (118) was solubilized in dimethyl sulfoxide at a concentration of 10 ppm. At 1 and 2 days after PCB (118) injection (100 ng/g body weight), both osteoclastic and osteoblastic activities, and plasma calcium levels were measured. In an in vitro study, then, both osteoclastic and osteoblastic activities as well as each marker mRNA expression were examined. At 2 days, scale osteoclastic activity in PCB (118)-injected goldfish increased significantly, while osteoblastic activity did not change significantly. Corresponding to osteoclastic activity, plasma calcium levels increased significantly at 2 days after PCB (118) administration. Osteoclastic activation also occurred in the marker enzyme activities and mRNA expressions in vitro. Thus, we conclude that PCB (118) disrupts bone metabolism in goldfish both in vivo and in vitro experiments.     

Trans-Pacific dispersal of loggerhead turtle hatchlings inferred from numerical simulation modeling

We used Lagrangian numerical simulations to examine the trans-Pacific dispersal processes of loggerhead turtle hatchlings. Ten thousand simulated particles were released from each of the three nesting regions in Japan and tracked for 5 years. Results showed many particles moving eastward, drifting in the Kuroshio Current followed by the Kuroshio Extension Current. However, no particles reached Baja California, a known feeding area, through passive processes, indicating that trans-Pacific transportation requires active swimming by turtles. The duration of the trans-Pacific dispersal was estimated to be at least 1.6–3.4 years, with some turtles drifting in the Kuroshio Countercurrent and remaining in the western Pacific even after 5 years. This indicates that as revealed by previous genetic studies, not all loggerheads always disperse along a trans-Pacific route. The findings showed that survival and expected growth rates varied widely according to ambient temperatures during drifting, which in turn depended on nesting location.     

Growth and grazing rates of the tintinnid ciliate<Emphasis Type="Italic"> Favella taraikaensis</Emphasis> on the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis>

Growth and feeding activities of the tintinnid ciliate Favella taraikaensis fed the toxic dinoflagellate Alexandrium tamarense were examined in laboratory experiments. Both growth and ingestion rates of F. taraikaensis as a function of the A. tamarense concentration were fitted to a rectangular hyperbolic equation. The maximum growth and ingestion rates of F. taraikaensis were 1.0 day^–1 and 2.8 cells ind. h^–1 (carbon specific ingestion rates: 3.5 day^–1), respectively, which are both included in the range of previous data reported for Favella spp. feeding on other algae. The gross growth efficiency (GGE) of F. taraikaensis ranged from 0.26 to 0.49 (mean value 0.40) at the concentration of 10–800 cells ml^–1, which is within the range of previous data on Favella spp. Also, the growth and ingestion rates and GGE of F. taraikaensis on A. tamarense were not significantly different from the values on another non-toxic dinoflagellate (Heterocapsa triquetra) at two different prey concentrations. This indicates that the toxicity of A. tamarense probably did not influence the feeding and growth activities of F. taraikaensis at concentrations of less than ca. 800 cells ml^–1. To evaluate the grazing by F. taraikaensis on A. tamarense blooms in the field, the population dynamics of A. tamarense were simulated based on the growth and ingestion parameters of F. taraikaensis. As a result, the grazing impact by F. taraikaensis was considered to potentially regulate the development of A. tamarense blooms. If the toxicity of A. tamarense does not influence the growth and feeding activities of F. taraikaensis, the occurrence of such grazer plankton are considered to be important for predicting the course of A. tamarense bloom dynamics under natural conditions.Communicated by T. Ikeda, Hakodate     

Recycling process for yttria-stabilized tetragonal zirconia ceramics using a hydrothermal treatment

The recycling process for 3 mol% yttria-stabilized tetragonal zirconia polycrystals (3Y-TZP) sintered at 1450°–1550°C was examined by applying low-temperature degradation of zirconia ceramics under hydrothermal conditions. Hydrothermal treatment at a temperature from 200° to 240°C can lead to the spontaneous disintegration of 3Y-TZP sintered bodies into powdery particles. The hydrothermally obtained zirconia powder was found to consist of primary particles and aggregated particles. Detailed X-ray diffraction measurement revealed the formation of a cubic zirconia phase in the 3Y-TZP sintered bodies, which seemed to inhibit the disintegration of aggregated particles toward the primary particle level. The reclaimed 3Y-TZP powder was sintered again through a conventional powder processing route. The mechanical properties and microstructure of recycled 3Y-TZP sintered specimens were examined by comparison with those of the original 3Y-TZP sintered bodies. Dense recycled 3Y-TZP sintered at a higher temperature exhibited higher fracture toughness to some degree than the original 3Y-TZP.     

Verification of the silica deficiency hypothesis based on biogeochemical trends in the aquatic continuum of Lake Biwa-Yodo River-Seto Inland Sea, Japan

The silica deficiency hypothesis holds that increases of still waters caused by hydraulic alterations and high nitrogen (N) and phosphorus (P) discharges enhance the growth of freshwater diatoms, which take up the dissolved silicate (DSi) supplied by natural weathering. The consequent decrease in the DSi supply to the sea is advantageous to flagellates (nonsiliceous and potentially harmful) but not to diatoms (siliceous and mostly benign) in coastal marine ecosystems. Verification of this hypothesis has been hampered by lack of relevant data, particularly in Asia. We investigated the aquatic continuum composed of Lake Biwa, the Yodo River, and the Seto Inland Sea, Japan, where the natural conditions make the silica deficiency less likely to emerge due to the inherently rich supply of DSi. The results showed that the silica was retained both in the lake and nearby the estuary. The relative dominance of diatom and flagellates could not be explained solely by the stoichiometric arguments but by the supportive discussion on the difference of their behavioral characteristics and the process nearby the estuary, where direct inputs of N and P and effluent Si enhanced diatom bloom, even though the Si/N ratio was lowered in the upstream reservoir. Thus the retention of DSi occurred in two places: in the lake and nearby the estuary, where the other N and P are loaded directly. The rate of DSi retention correlated with socio-economic changes, such as rapid economic growth in the 1960s and mitigations implemented after the 1980s. Sensitivity of this continuum to the Si processes suggests the global significance of this hypothesis.     

A spatially explicit empirical model of structural development processes in natural forests based on climate and topography

Stand structure develops with stand age. Old-growth forests with well-developed stand structure support many species. However, development rates of stand structure likely vary with climate and topography. We modeled structural development of 4 key stand variables and a composite old-growth index as functions of climatic and topographic covariates. We used a hierarchical Bayesian method for analysis of extensive snap-shot National Forest Inventory (NFI) data in Japan (n = 9244) to account for differences in stand age. Development rates of structural variables and the old-growth index exhibited curvilinear responses to environmental covariates. Flat sites were characterized by high rates of structural development. Approximately 150 years were generally required to attain high values (approximately 0.8) of the old-growth index. However, the predicted age to achieve specific values varied depending on environmental conditions. Spatial predictions highlighted regional variation in potential structural development rates. For example, sometimes there were differences of >100 years among sites, even in the same catchment, in attainment of a medium index value (0.5) after timber harvesting. The NFI data suggested that natural forests, especially old natural forests (>150 years), remain generally on unproductive ridges, steep slopes, or areas with low temperature and deep snow, where many structural variables show slow development rates. We suggest that maintenance and restoration of old natural forests on flat sites should be prioritized for conservation due to the likely rapid development of stand structure, although remaining natural forests on low-productivity sites are still important and should be protected.     

Polychlorinated-naphthalenes, -biphenyls, -dibenzo-p-dioxins, -dibenzofurans and p,p'-DDE in bluefin tuna,swordfish, cormorants and barn swallows from Italy

Concentrations of p,p'-DDE, polychlorinated biphenyl congeners (PCBs), polychlorinated-dibenzo-p-dioxins (PCDDs), -dibenzofurans (PCDFs) and -naphthalenes (PCNs) were measured in bluefin tuna, swordfish, common cormorants, and barn swallows collected from Italy. Average concentrations of PCBs in livers of tuna, swordfish, cormorant, and swallows were 930, 745, 1420 and 1230 ng PCBs g-1, w.w. respectively. p,p'-DDE was found in tuna, swordfish, cormorant, and swallow livers at mean concentrations of 82, 135, 166 and 95 ng DDE g-1, w.w. respectively. PCNs were found in all the samples analyzed, although at concentrations less than those reported for biota from the Baltic Sea. PCBs, particularly, non-ortho coplanar PCBs accounted for 80-90% of the total TEQs in tuna and swordfish. Relative contribution of PCDDs/DFs to TEQs was greater in cormorants and swallows compared to that in fishes. PCDD/DF congeners accounted for up to 80 and 45% of the total TEQs in cormorants and swallows, respectively.     

Concentrations of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like polychlorinated biphenyls in blood and breast milk collected from 60 mothers in Sapporo City, Japan

We measured the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), non-ortho polychlorinated biphenyls (non-ortho PCBs), and mono-ortho polychlorinated biphenyls (mono-ortho PCBs) in paired samples of blood and breast milk collected from 60 mothers in Sapporo City, Hokkaido Prefecture, Japan. The present study is one of the few studies in which PCDDs, PCDFs, and dioxin-like PCBs have been measured in blood and breast milk collected from 60 same mothers. Of these 60 mothers, 30 were primipara (mean: 30.1 years, median: 28.0 years) and 30 were multipara (mean: 32.2 years, median: 32.5 years). The arithmetic mean TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in blood and breast milk of the primiparous mothers were 9.0, 3.3, 5.7, and 0.4 pg TEQ g(-1) lipid, respectively, and 5.2, 2.2, 4.5, and 0.4 pg TEQ g(-1) lipid, respectively, with the total TEQ concentrations of these dioxin-like compounds being 9.3-42.9 (mean: 18.4, median: 17.3) and 7.0-41.1 (mean: 12.3, median: 11.4) pg TEQ g(-1) lipid, respectively. In the case of multiparous mothers, the arithmetic mean TEQ concentrations of these dioxin-like compounds in blood and breast milk were 7.1, 2.7, 5.3, and 0.4 pg TEQ g(-1) lipid, respectively, and 3.9, 1.7, 3.8, and 0.4 pg TEQ g(-1) lipid, respectively, with the total TEQ concentrations being 3.4-28.1 (mean: 15.5, median: 13.9) and 2.7-20.0 (mean: 9.8, median: 9.2)pg TEQ g(-1) lipid, respectively. The total TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in blood and breast milk of primiparous mothers in Sapporo City appeared to be generally lower than those recently surveyed throughout the greater area of Japan. Significant correlations were observed between age and the total TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in the blood of primiparae and multiparae, and significant correlations were also observed between age and the total TEQ concentrations of these dioxin-like compounds in the breast milk of primiparae and multiparae. The total TEQ concentrations of PCDDs, PCDFs, and dioxin-like PCBs in the blood of primiparous mothers showed a close correlation to those in their breast milk and also showed good correlations between the total TEQ concentrations of these dioxin-like compounds in the blood of multiparous mothers and those in their breast milk. The result of a comparison of the arithmetic mean concentrations of each congener of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in blood and breast milk indicated that the transfer of octachlorodibenzo-p-dioxin (OCDD) from the blood to the breast milk was lower than those of other congeners. In contrast, among mono-ortho PCBs congeners, the arithmetic mean concentrations of 2,3',4,4',5-pentachlorobiphenyl (PentaCB) (#118) and 2,3,3',4,4'-PentaCB (#105) in the breast milk were slightly higher than those in the blood, which suggested that breast-fed infants may be at higher risk from mono-ortho PCBs.