Introduction of Yeast Metallothionein Gene (CUP1) into Plant and Evaluation of Heavy Metal Tolerance of Transgenic Plant at the Callus Stage

For phytoremediation, it is necessary to use plants with a large biomass and ability to absorb and accumulate heavy metals. Attempts were made to produce heavy metal-tolerant plants by introducing a metallothionein gene, CUP 1, into plants with a large biomass. In the present study, we evaluated the heavy metal tolerance of sunflower plants at the callus stage in terms of 2,3,5-triphenyl-tetrazolium chloride (TTC)-reducing activity, measured the mRNA expression level of the introduced gene, and selected tolerant clones. The transgenic calli of several strains showed a high TTC-reducing activity even after treatment with Cd 200 μM for 10 d. In the transgenic calli of a few strains, which showed a low TTC-reducing activity, the amount of metallothionein was lower than that in the other transgenic strains. Conversely, some transgenic calli with a high TTC-reducing activity expressed a much higher amount of mRNA of the introduced gene.     

Intrinsic Biodegradation of Heavy Oil from Nakhodka and the Effect of Exogenous Fertilization at a Coastal Area of the Sea of Japan

We performed a field experiment in thebiodegradation of heavy oil spilled from the Russian tankerNakhodka on a beach in the Sea of Japan. We collectedoil-contaminated cobbles and treated half with nitrogen andphosphorus slow-release fertilizers to stimulate microbialdegradation of the oil; the other half acted as unfertilizedcontrols. The cobbles were placed in porous acrylic vessels andsubmerged. We monitored changes in the oils, macronutrients,microbial community structure and amount of chlorophyll a. There were no significant differences in these criteriabetween the fertilized and unfertilized vessels, apart from anincrease in chlorophyll a in the fertilized vessels.However, there was a major intrinsic degradation of semi-volatile oil compounds in the unfertilized vessels; this occurred at a rate similar to that in the fertilized vessels, despite the low concentration of macronutrients in the seawater at the site.     

Induction of short-term pseudopregnancy in gilts by the administration of estradiol benzoate or estradiol dipropionate to achieve ovulatory synchronization for embryo collection

A study was conducted to investigate whether ovulation in gilts could be synchronized for embryo collection by the administration of estradiol benzoate (EB) or estradiol dipropionate (EDP) to induce pseudopregnancy, followed by the treatment with prostaglandin F_2α (PGF_2α) on 10 days after. Ten gilts each received a total of 20 mg of EB or EDP on Day 10 or EB on Day 10 and 14 to induce pseudopregnancy (Day 0 = onset of estrus). Donors received PGF_2α 10 or 15 days (as a control) after the first administration of estrogens and subsequently eCG and hCG, and were then inseminated artificially. The embryos were collected 7 days after the administration of hCG, and assessed for embryo yield and their developmental stages. All protocols resulted in good embryo yield (9.8–13.2 embryos in average), and the embryos showed average ability to develop to the expanded blastocyst stage (3.29–4.03 as developmental scores) without any significant differences among the protocols. These results suggest that the administration of PGF_2α 10 days after the treatment of gilts with EB or EDP would allow synchronization of ovulation and embryo collection, as well as shortening the period from estrus detection to embryo collection, thus improving embryo collection efficiency.     

Excess polyspermy reduces the ability of porcine oocytes to promote male pronuclear formation after in vitro fertilization

Male pronucleus (MPN) formation is a very important physiological event during fertilization, which affects in vitro production of transferrable embryos. The aim of this study was to find out the correlation between the number of penetrated sperm and the occurrence of failure of MPN formation in porcine oocytes. In vitro matured porcine oocytes were fertilized in vitro with frozen epididymal sperm. Two different frozen sperm lots were tested in this study, which were different in terms of polyspermy rates. The numbers and the status of penetrated sperm in oocytes were evaluated 10 h after insemination. Under high polyspermy condition, the polyspermy rate was 83.5% with an average mean of 3.5 sperms per penetrated oocyte, whereas the percentage of polyspermy was 65.5% with an average mean of 2.4 sperms per penetrated oocyte under moderate polyspermic condition. Correlation analysis revealed a negative correlation between the number of penetrated sperm and their MPN formation percentage both in the sperm lot of high polyspermy (R = −0.560, p < 0.05) and in the sperm lot of moderate polyspermy (R = −0.405, p < 0.05) which suggests that penetration of excessive spermatozoa disables the oocyte cytoplasm to promote MPN formation.     

Absorption of 6-O-caffeoylsophorose and its metabolites in Sprague-Dawley rats detected by electrochemical detector-high-performance liquid chromatography and electrospray ionization-time-of-flight-mass spectrometry methods

Absorption and metabolism of a natural compound, 6-O-caffeoylsophorose (CS) from acylated anthocyanins in a red vinegar fermented with purple sweet potato, were clarified. The absorption of CS and conjugated CS in blood from orally administrated Sprague-Dawley rats at a dose of 400 mg/kg was investigated by electrochemical detection-high performance liquid chromatography. As a result, CS was successfully detected in rat plasma (AUC(0-6h), 108.6 ± 8.1 nmol h/mL) and was found to be an intact absorbable polyphenol. In addition, half of the absorbed CS was detected as its conjugates (AUC(0-6h), 50.7 ± 5.7 nmol h/mL) as well as caffeic and ferulic acids from CS. By a time-of-flight-mass spectrometric analysis of CS-administered plasma sample, glucuronide and methylated conjugates of CS were identified, in addition to glucuronide, methylated, or sulfate conjugates of caffeic and ferulic acids. Consequently, CS was absorbed in intact form into rat blood and partly degraded to caffeic and ferulic acids or metabolized by glucuronidation, methylation, or sulfatation.     

Recombinant interferon enhances monoclonal antibody-targeting of carcinoma lesions in vivo

Heterogeneity in the expression of tumor-associated antigens, as defined by the binding of monoclonal antibodies, is a characteristic common to most, if not all, human carcinoma cell populations. Antigen-negative cells within the population can escape detection and therapy by their failure to bind the appropriate antibody. Therefore, the extent of antigenic heterogeneity is an important consideration when designing protocols for the management of cancer by administration of monoclonal antibodies. One approach to counteracting the effect of antigenic heterogeneity is the use of clone A of recombinant human leukocyte interferon (Hu-IFN-alpha A). Administration of Hu-IFN-alpha A in vivo effectively increased the amount of tumor antigen expressed by a human colon xenograft in situ and augmented the localization of a radiolabeled monoclonal antibody to the tumor site. Concomitant administration of Hu-IFN-alpha A and monoclonal antibody may thus be effective in overcoming the antigenic heterogeneity of carcinoma cell populations and in enhancing the efficacy of monoclonal antibodies in the detection and treatment of carcinoma lesions.     

A mechanism of lithium storage in disordered carbons

High-resolution electron microscopy and lithium-7 nuclear magnetic resonance measurements were carried out for a disordered carbon material, prepared by heat treatment of polyphenylene, in which lithium was stored electrochemically. The nuclear magnetic resonance spectrum suggests the existence of Li(2) covalent molecules in the carbon material. This extra covalent site of lithium storage promises extraordinarily high energy density for secondary batteries.     

Soil hardness regulates the root penetration by trees planted on anthropogenic growing bases in coastal forests in Japan: new endeavors to reforest the coastal disaster prevention forests with high resilience for tsunami

Journal of Soils and Sediments - To restore the disaster resilience of coastal forests in tsunami-prone areas, growing bases have been artificially constructed with embanking before afforestation...     

Studies on nitrogen metabolism in the withered leaves of sugar beets in warmer region of Japan

We have many studies (1, 2) on the growing process of sugar beets. The results 3) of the examinations in the past have hown that among the three clements nitrogen tas the greatest influence on the growth and ield. Therefore many studies (4, 5) of nutrient metabolism arc carried out laying tress on nitrogen.     

Isolation and characterization of a novel Arabidopsis thatiana mutant that requires a high concentration of boron

We isolated a novel Arabidopsis thaliana mutant line that requires high levels of boron (B) for normal growth. Line 8–21 was identified from ethyl methanesulfonate-mutagenized M2 population. When grown in medium containing 3 μm or less B in the form of boric acid, the fresh weights of aerial portions of the mutant were about a half of those of the wild type, but in that containing 300 μm B, the growth appeared normal. The mutant plants did not shown any difference in root growth from the wild-type plants in the range of B concentrations tested. When grown with 30 μm B, the B concentration in shoots of the line 8–21 was similar to that in the wild-type, suggesting that the mutant could not utilize B efficiently. Line 8–21 was not allelic to bor1-1 (Noguchi et al., Plant Physiol., 115, 901–906, 1997). A significant portion of F2 plants from the crosses between the wild-type and the mutant grew poorly on a low B media, suggesting segregation of the mutation.