Simple method to reduce interference from excess magnesium in cadmium immunoassays

In order to develop a rapid inexpensive test for cadmium in rice, we identified an antibody specific for cadmium-EDTA complexes; this antibody binds to cadmium-EDTA with a Kd of approximately 10(-8) M. Although the antibody's cross reactivity to magnesium was minimal (Kd approximately 10(-5) M), the high toxicity of cadmium coupled with the high natural occurrence of magnesium in rice resulted in a situation where magnesium interfered with cadmium determination and resulted in falsely elevated estimates of cadmium. Fortunately, the formation constant of EDTA for cadmium is approximately 5 x 10(7) times higher (at pH 7) than the formation constant of EDTA for magnesium, and we were able to eliminate the magnesium interference by judicious selection of the EDTA concentration used in the assay. The resulting equilibria are complex, but we show that a relatively simple two-step model in which cadmium and magnesium compete for EDTA followed by cadmium-EDTA and magnesium-EDTA competing for antibody provided a good fit to the measured data. These analyses enabled appropriate selection of the optimum EDTA concentration for an immunoassay with improved selectivity.     

Ontogenetic habitat shift in <Emphasis Type="Italic">Pugettia quadridens</Emphasis> on the coast of Sagami Bay,Japan

Population dynamics of the kelp crab Pugettia quadridens, a potential predator of juvenile abalone, were investigated on the coast of Nagai, Sagami Bay, to examine its cryptic ontogenetic habitat shift. Monthly quantitative sampling over 2 years, which was carried out in 11 different subtidal habitats (< 0.1–8 m deep), revealed that P. quadridens changes ontogenetically according to its habitat: P. quadridens occurred in shallow lower subtidal (1–4 m) small red algal turfs (mainly Grateoloupia cornea) almost all year around, when the population was dominated by juveniles and stage I (immature) individuals; on the other hand, in the shallow upper subtidal (0.1–1 m) habitat (mainly Sargassum fusiforme beds near the low tidal mark), P. quadridens occurred only during winter to early spring, when the population was dominated by individuals of senior ontogenetic stages. These finding suggest that P. quadridens inhabit subtidal small red algal turfs during their early benthic phase, and then, the majority of larger individuals of stages II and III migrate to Sargassum beds. Our results suggest that the impact of P. quadridens predation on abalone stocks is limited in the present study site, primarily because the habitat of P. quadridens is segregated from that of juvenile abalone.     

A pubescence color gene enhances tolerance to cold-induced seed cracking in yellow soybean

In yellow soybean, severe cold weather causes seed cracking on the dorsal side. Yellow soybeans carry the I or iⁱ allele of the I locus and have a yellow (I) or pigmented (iⁱ) hilum. We previously isolated an additional allele, designated as Ic, of the I locus, and reported that yellow soybeans with the IcIc genotype may be tolerant to cold-induced seed cracking. The Ic allele by itself, however, does not confer high tolerance. The association of a pubescence color gene (T) with suppression of low-temperature-induced seed coat deterioration has been previously reported. In the present study, we tested whether T is effective for the suppression of cold-induced seed cracking using two pairs of near-isogenic lines for the T locus in the iⁱiⁱ or IcIc background. In both backgrounds, the cracked seed rate of the near-isogenic line with the TT genotype was significantly lower than that with the tt genotype, which indicates that T has an inhibitory effect on cold-induced seed cracking. Furthermore, we also showed that gene pyramiding of Ic and T can improve tolerance to cold-induced seed cracking. Our findings should aid the development of highly SC-tolerant cultivars in soybean breeding programs.     

Inhibition of hepatitis C virus replication and viral helicase by ethyl acetate extract of the marine feather star Alloeocomatella polycladia

Hepatitis C virus (HCV) is a causative agent of acute and chronic hepatitis, leading to the development of hepatic cirrhosis and hepatocellular carcinoma. We prepared extracts from 61 marine organisms and screened them by an in vitro fluorescence assay targeting the viral helicase (NS3), which plays an important role in HCV replication, to identify effective candidates for anti-HCV agents. An ethyl acetate-soluble fraction of the feather star Alloeocomatella polycladia exhibited the strongest inhibition of NS3 helicase activity, with an IC(50) of 11.7 μg/mL. The extract of A. polycladia inhibited interaction between NS3 and RNA but not ATPase of NS3. Furthermore, the replication of the replicons derived from three HCV strains of genotype 1b in cultured cells was suppressed by the extract with an EC(50) value of 23 to 44 μg/mL, which is similar to the IC(50) value of the NS3 helicase assay. The extract did not induce interferon or inhibit cell growth. These results suggest that the unknown compound(s) included in A. polycladia can inhibit HCV replication by suppressing the helicase activity of HCV NS3. This study may present a new approach toward the development of a novel therapy for chronic hepatitis C.     

Current status of tropical fruit breeding and genetics for three tropical fruit species cultivated in Japan: pineapple,mango, and papaya

Tropical fruit crops are predominantly produced in tropical and subtropical developing countries, but some are now grown in southern Japan. Pineapple (Ananas comosus), mango (Mangifera indica) and papaya (Carica papaya) are major tropical fruits cultivated in Japan. Modern, well-organized breeding systems have not yet been developed for most tropical fruit species. Most parts of Japan are in the temperate climate zone, but some southern areas such as the Ryukyu Islands, which stretch from Kyushu to Taiwan, are at the northern limits for tropical fruit production without artificial heating. In this review, we describe the current status of tropical fruit breeding, genetics, genomics, and biotechnology of three main tropical fruits (pineapple, mango, and papaya) that are cultivated and consumed in Japan. More than ten new elite cultivars of pineapple have been released with improved fruit quality and suitability for consumption as fresh fruit. New challenges and perspectives for obtaining high fruit quality are discussed in the context of breeding programs for pineapple.     

Locally synchronized synaptic inputs

Synaptic inputs on dendrites are nonlinearly converted to action potential outputs, yet the spatiotemporal patterns of dendritic activation remain to be elucidated at single-synapse resolution. In rodents, we optically imaged synaptic activities from hundreds of dendritic spines in hippocampal and neocortical pyramidal neurons ex vivo and in vivo. Adjacent spines were frequently synchronized in spontaneously active networks, thereby forming dendritic foci that received locally convergent inputs from presynaptic cell assemblies. This precise subcellular geometry manifested itself during N-methyl-D-aspartate receptor-dependent circuit remodeling. Thus, clustered synaptic plasticity is innately programmed to compartmentalize correlated inputs along dendrites and may reify nonlinear synaptic integration.     

Spontaneous Adenosquamous Carcinoma with Rapid Growth and EMT-like Changes in the Mammary Gland of a Young Adult Female BALB/c Mouse

This study histopathologically and immunohistochemically investigated a spontaneously occurring single mass subcutaneously located in the left lower abdomen of a female BALB/cAJcl−nu/+ mouse at 10 weeks of age. The mass was about 20 × 15 × 10 mm in size after formalin fixation; nevertheless, it was not detected by clinical observations at 9 weeks of age. H&E staining revealed the tumor origin was epithelial and probably arose from the mammary gland, and the tumor cells demonstrated a squamous, acinar or polyhedral/basal pattern. A cell kinetics analysis revealed that many of the tumor cells of the squamous, acinar or polyhedral/basal component were positive for PCNA and cyclin D1, although there were a few of TUNEL-positive tumor cells in all of the components. An epithelial/mesenchymal analysis demonstrated that most of the tumor cells of the squamous and acinar components contained keratin and E-cadherin; however, most of the tumor cells of the polyhedral/basal component were less or very weakly positive for these markers. The tumor cells of the squamous component were negative for vimentin and SMA; however, many of the tumor cells of the polyhedral/basal component exhibited vimentin. In addition, expression of SMA was confirmed in some tumor cells of the acinar and basal components. Based on the microscopic and immunohistochemical characterizations, the tumor was diagnosed to be adenosquamous carcinoma that originated from the mammary gland with rapid growth, and the tumor cells demonstrated epithelial-mesenchymal transition-like changes.     

Pole and global shape of 25143 Itokawa

The locations of the pole and rotation axis of asteroid 25143 Itokawa were derived from Asteroid Multiband Imaging Camera data on the Hayabusa spacecraft. The retrograde pole orientation had a right ascension of 90.53 degrees and a declination of -66.30 degrees (52000 equinox) or equivalently 128.5 degrees and -89.66 degrees in ecliptic coordinates with a 3.9 degrees margin of error. The surface area is 0.393 square kilometers, the volume is 0.018378 cubic kilometers with a 5% margin of error, and the three axis lengths are 535 meters by 294 meters by 209 meters. The global Itokawa revealed a boomerang-shaped appearance composed of two distinct parts with partly faceted regions and a constricted ring structure.     

Efficient digestion and structural characteristics of cell walls of coffee beans

Screening of effective food-processing cellulase for digestion of cell walls of coffee beans was carried out, and the cellulase from Trichoderma sp. was selected. The digestion of the cell walls of green and roasted coffee beans was carried out by sequential procedures of alkali boiling (0.1 M Na2CO3 buffer, pH 10, and 0.1 M NaOH), cellulase digestion, autoclaving with 0.1 M NaOH, and cellulase redigestion. The total digestion yields were >95 and >96%, respectively. The cell walls became thin, and the final residues of the cell walls were easily broken into small pieces. The neutral sugar analysis of the digestion or the extract and the residues and the microscopy observations with staining with toluidine blue O, Yariv reagent, and calcofluor for the residue in each step were investigated. Four structures, the galactomannan-cellulose (center part), the membrane of the arabinogalactan protein, the cellulose-rich galactomannan layer, and the arabinogalactan protein-rich layers (outer part), were found in the cell walls.     

Groundwater Deterioration Caused by Induced Recharge: Field Survey and Verification of the Deterioration Mechanism by Stochastic Numerical Simulation

Our field survey showed that thequality of shallow groundwater around the KatsuraRiver in the Kyoto Basin was strongly affected by theinfiltration of river water. Furthermore, that thedeterioration of the groundwater in the southern areato the west of the Katsura River may be related to theincrease in groundwater extraction. To clarify themechanism of groundwater deterioration, we havedeveloped a stochastic method to simulate groundwaterflow. The results showed that there was a largereduction in the groundwater level where groundwaterextraction was intense and recharge flowed from theKatsura River to the high extraction areas in thesouthern region. Another simulation showed that if thegroundwater extraction was 10% of the present removalrate, there would be little recharge from the KatsuraRiver into the groundwater and the quality of thegroundwater would be improved. Thus, we conclude thatthe cause of groundwater deterioration is probably dueto the induced recharge of deteriorated river waterfrom the Katsura River.