Purification of gonadotropins (PmGTH I and II) from red seabream (Pagrus major) and development of a homologous radioimmunoassay for PmGTH II

Two gonadotropic glycoproteins (PmGTH I and II) were purified by ion-exchange chromatography, gel filtration and preparative SDS-PAGE, from pituitaries of red seabream, a marine teleost which has an asynchronous-type ovary and spawns almost daily during the spawning season. The glycoproteins were composed of distinct subunits and the molecular weights were estimated to be 32 and 38 kDa for PmGTH I and PmGTH II, respectively. Both PmGTH I and II were active in two homologous bioassays: in vitro oocyte maturation and/or in vitro estradiol-17 production assays. These two GTHs were distinct in electrostatic properties, molecular weight, stability and yields from pituitaries during the spawning season. These properties suggest that PmGTH I and II correspond to salmon GTH I and II, respectively.A homologous radioimmunoassay with which to measure PmGTH II was developed using a rabbit antiserum against the subunit of PmGTH II and intact PmGTH II as standards and radioactive competitors. Competition curves for red seabream plasma and pituitary extract were parallel to the standard curve, while PmGTH I had low cross-reactivity (3.1 %) with the antibody. This specific RIA system showed an in vivo LHRHa induced GTH surge in the plasma of female red seabream.     

Discrimination of excellent-grade ‘nori’, the dried laver Porphyra spp., with analytical methods for volatile compounds

Analytical methods were investigated for the discrimination of ‘nori’, the dried laver Porphyra spp. sheet products, which were organoleptically judged as excellent-grade for commercial distribution. Volatile compounds of nori were analyzed with an electronic nose and a gas chromatography-mass spectrometer (GC/MS) with headspace solid-phase micro-extraction. Each chopped dry nori sample was sealed in a vial and the headspace gas at 40 °C was analyzed. Excellent-grade nori product of ‘Akime’, first autumn shoots, of one season could be discriminated from other products by using principal component analyses (PCA) for the electronic nose response. PCA using a peak area of 32 compounds detected in the GC/MS analysis clearly discriminated all excellent-grade samples from all other samples. Excellent-grade products were considered to be characterized by higher total contents of 6-methyl-5-hepten-2-one, octanal and trimethylamine, and by lower total volatile contents, especially 1-pentanol, 2-pentenal, (E)-2-hexenal, acetoin, 2-methyl-2-butenal, 3-methyl butanal and others. These findings revealed the possibility of analytical discrimination of excellent-grade nori products as a substitution for organoleptic testing.     

Spontaneous hemangiosarcoma in the spleen and liver of a young rat

Spontaneous hemangiosarcoma in young rats is rare. In this report, we describe a case of a spontaneous hemangiosarcoma in the spleen and liver of young rats. At necropsy, multiple pale red masses were observed in the spleen. Histopathologically, solid growth and haphazardly arranged neoplastic cells were observed, although no characteristic growth pattern was observed. In contrast, irregularly sized small slit-shaped spaces containing erythrocytes were found among the neoplastic cells. Reticular fibers incompletely surrounding the neoplastic cells were observed by silver staining. Immunohistochemistry revealed that the neoplastic cells were positive for vWF and CD34. Electron microscopic examination revealed that the neoplastic cells had erythrocytes in the lumen and Weibel-Palade bodies in the cytoplasm and were arranged along a discontinuous basal lamina. These features indicate that the tumor originated from vascular endothelial cells. Based on these results, the tumor was diagnosed as a hemangiosarcoma in the spleen and liver.     

Occurrence of Pacific bluefin tuna Thunnus orientalis larvae off the Pacific coast of Tohoku area,northeastern Japan: Possibility of the discovery of the third spawning ground

Recently, based on the histological studies it is suggested that Pacific bluefin tuna (PBF) spawns in the Kuroshio–Oyashio transition, off the Pacific coast of northeastern Japan, in addition to the previously identified two spawning areas of the northwestern Pacific around the Nansei Islands and the Sea of Japan. Distributional surveys for PBF larvae have not been conducted in the Kuroshio–Oyashio transition, and thus, we conducted PBF larval surveys at twenty stations in this area in early August 2018 to obtain evidence for the spawning of PBF. Twelve PBF larvae (3.9–7.2 mm in body length) were collected at six stations where sea surface temperature ranged from 27.2 to 28.2°C. Otolith microstructure analysis indicated that age of these larvae ranged from 4 to 11 days after hatching and the larvae hatched in late July. Growth of the collected larvae was comparable to those in the other two spawning grounds. Therefore, PBF spawns, hatches, and at least survives to the postflexion stage, 11 days after hatching in the Kuroshio–Oyashio transition. This fact potentially has a large impact on recruitment processes of PBF if they survive to recruitment in this third spawning ground.     

Validation of daily increment formation in otoliths of immature and adult Japanese anchovy <Emphasis Type="Italic">Engraulis japonicus</Emphasis>

In order to validate daily increment formation in otoliths of immature and adult Japanese anchovy Engraulis japonicus, three rearing experiments using chemical marking of otoliths were conducted on adult anchovy in summer 2004 and immature anchovy in summer 2005 and in winter 2006. In the two experiments conducted in summer, the number of otolith microincrements between alizarin complexone (ALC) marks showed that microincrements were formed daily. In the summer 2005 experiment, immature anchovy under conditions of reduced daily food rations also showed daily microincrement formation. Average increment width was 0.9 μm in adults and 1.8–3.1 μm in immature anchovy. In contrast, no clear increments were observed between ALC marks on the otoliths from the experiment in winter 2006, and scanning electron microscope (SEM) observations failed to confirm clear increment formation. We consider that low water temperatures (<13–14°C) restricted otolith growth and lowered the contrast between the discontinuous and the incremental zones of the otolith increments. For age estimation of Japanese anchovy, clear increments wider than about 1 μm in the otolith can be regarded as daily increments. However, daily age estimation of immature and adult anchovy that experience low water temperatures in winter may be difficult due to the obscurity of the increments.     

Surface analysis of paper containing polymer additives by X-ray photoelectron spectroscopy I: Application to paper containing dry strength additives

The surfaces of handsheets containing polydiallyl-dimethylammonium chloride (PDADMAC) and anionic polyacrylamide (A-PAM) were analyzed using X-ray photoelectron spectroscopy (XPS). The observed N1s chemical shifts were compared with those of the strength additives. Chemical covalences on paper surfaces due to a small amount of polymer additives were clarified and determined by an XPS curve-fitting technique. Some of the problems associated with surface analysis of paper by XPS are discussed. The effects of strength additives on the fiber surface or between fibers are illustrated. This technique appears promising as a tool to analyze paper surfaces treated with small amounts of polymer additives.Part of this paper was presented at the 64th Pulp and Paper Research Conference, Tokyo, Japan, June 19–20, 1997     

Spatial structures of N2O, CO2, and CH4 fluxes from Acacia mangium plantation soils during a relatively dry season in Indonesia

We investigated spatial structures of N₂O, CO₂, and CH₄ fluxes during a relatively dry season in an Acacia mangium plantation stand in Sumatra, Indonesia. The fluxes and soil properties were measured at 1-m intervals in a 1 × 30-m plot (62 grid points) and at 10-m intervals in a 40 × 100-m plot (55 grid points) at different topographical positions of the upper plateau, slope, and valley bottom in the plantation. Spatial structures of each gas flux and soil property were identified using geostatistical analysis. The means (±SD) of N₂O, CO₂, and CH₄ fluxes in the 10-m grids were 0.54 (±0.33) mg N m⁻² d⁻¹, 2.81 (±0.71) g C m⁻² d⁻¹, and −0.84 (±0.33) mg C m⁻² d⁻¹, respectively. This suggests that A. mangium soils function as a larger source of N₂O than natural forest soils in the adjacent province on Sumatra during the relatively dry season, while CO₂ and CH₄ emissions from the A. mangium soils were less than or consistent with those in the natural forest soils. Multiple spatial dependence of N₂O fluxes within 3.2 m (1-m grids) and 35.0 m (10-m grids), and CO₂ fluxes within 1.8 m (1-m grids) and over 65 m (10-m grids) was detected. From the relationship among N₂O and CO₂ gas fluxes, soil properties, and topographic elements, we suggest that the multiple spatial structures of N₂O and CO₂ fluxes are mainly associated with soil resources such as readily mineralizable carbon and nitrogen in a relatively dry season. The soil resource distributions were probably controlled by the meso- and microtopography. Meanwhile, CH₄ fluxes were spatially independent in the A. mangium soils, and the water-filled pore space appeared to mainly control the spatial distribution of these fluxes.     

Site-level model–data synthesis of terrestrial carbon fluxes in the CarboEastAsia eddy-covariance observation network: toward future modeling efforts

Based on the model–data comparison at the eddy-covariance observation sites from CarboEastAsia datasets, we report the current status of the terrestrial carbon cycle modeling in monsoon Asia. In order to assess the modeling performance and discuss future requirements for both modeling and observation efforts in Asia, we ran eight terrestrial biosphere models at 24 sites from 1901 to 2010. By analyzing the modeled carbon fluxes against the CarboEastAsia datasets, the strengths and weaknesses of terrestrial biosphere modeling over Asia were evaluated. In terms of pattern and magnitude, the carbon fluxes (i.e., gross primary productivity, ecosystem respiration, and net ecosystem exchange) at the temperate and boreal forest sites were simulated best, whereas the simulation results from the tropical forest, cropland, and disturbed sites were poor. The multi-model ensemble mean values showed lower root mean square errors and higher correlations, suggesting that composition of multiple terrestrial biosphere models would be preferable for terrestrial carbon budget assessments in Asia. These results indicate that the current model-based estimation of terrestrial carbon budget has large uncertainties, and future research should further refine the models to permit re-evaluation of the terrestrial carbon budget.     

Labile pools of organic matter and microbial biomass in the surface soils under shifting cultivation in northern Thailand

In order to analyze the N mineralization process under shifting cultivation in northern Thailand, labile pools of soil organic matter were studied, which were considered to be the factors contributing to the N mineralization process. Organic C, (organic + NH₄ ⁺)-N, and hexose-C were extracted from fresh soils in the surface 0–5 cm layers with a 0.5 M K₂S₀. solution at 110°C in an autoclave (fraction A) or at room temperature with a reciprocal shaker (fraction B), and analyzed as labile pools of organic matter. In the traditional shifting cultivation system, the content of organic C in fraction A in the fallow fields for 8 to 15 y was 3,710 mg kg^-1 while that in the fallow fields for 1 y and 3 to 5 y was 2,640 and 2,600 mg kg^-1, respectively. A high correlation was observed between the contents of the labile pool in fraction A and total soil organic matter. The ratio of the pool in fraction A to total soil organic matter apparently remained constant through the input-output balance in the pool. The content of the labile pool in fraction B was the highest among the fields cultivated for 1 y after the slash and burn practice and it decreased in the course of the fallow period. The content of organic C was 548 mg kg^-1 in the fields cultivated for 1 y and 235 mg kg^-1 in the fallow fields for 8-15 y, respectively. There was a reverse relation between the contents of the pool in fraction B and microbial biomass. Therefore, the origin of the pool in fraction B was attributed to the microbial debris associated mainly with a decrease in the soil moisture content in the dry season. On the other hand, in the relatively intensive cultivation system, there was no significant difference in the contents of the labile pools both in fractions A and B among the land use stages, suggesting that the preservation mechanism of these pools, which was observed in the traditional cultivation system, did not operate well in the intensive system. In alternative farming systems in future, it will be essential to apply organic materials to soils to supply organic matter and to maintain the microbial biomass.