Recombinant human AhR-mediated GUS reporter gene assays for PCB congeners in transgenic tobacco plants in comparison with recombinant mouse and guinea pig AhRs

Four expression plasmids for recombinant human aryl hydrocarbon receptor (hAhR) consisting of a ligand binding domain of hAhR, a DNA-binding domain of LexA and a transactivation domain of VP16 as well as β-glucuronidase (GUS) reporter genes were constructed. All the expression plasmids were transformed into tobacco plants. The selected transgenic tobacco plants were used to assay. PCB congeners showed GUS activity in a TEF-dependent manner. The selected transgenic tobacco plant XhD4V17 was compared with the transgenic tobacco plants XmD4V26 and XgD2V23 containing recombinant mouse (m) AhR-mediated GUS reporter gene expression cassette and recombinant guinea pig (g) AhR-mediated GUS reporter gene expression cassette for PCB congener-inducible GUS activity. The data revealed that the tobacco plant XgD2V23 was the most active in PCB congener-inducible GUS activity. In a 1:1 mixture of PCB126 and PCB80 a reduced PCB126-induced GUS activity was observed in plant XgD2V23, which could possibly be due to interaction between PCB126 and PCB80.     

Gasification of waste plastics by steam reforming in a fluidized bed

The process of producing synthetic gas from waste plastics by steam reforming was investigated. To evaluate this process, the steam reforming of the oils derived from low-density polyethylene and polystyrene were carried out using a laboratory-scale fluidized bed of Ni-Al₂O₃ catalysts. The performance of gasification in terms of carbon conversion, gas yield, and gas compositions was examined. Although oils derived from plastics contain many kinds of heavy hydrocarbons and aromatics, they were well gasified at temperatures above 1023 K with a steam/carbon ratio of 3.5 and a weight hourly space velocity of 1 h⁻¹. The hydrogen content of the product gas was very high at approximately 72 vol% for polyethylene-derived oil and 68 vol% for polystyrene-derived oil. These compositions agreed well with the values calculated from chemical equilibrium.     

Body color change and serum steroid hormone levels throughout the process of sex change in the adult wrasse, <Emphasis Type="Italic">Pseudolabrus sieboldi</Emphasis>

Gonadal sex steroid hormones are the principal factors that directly control the gonadal and morphological alterations during sex change in hermaphrodite fish; however, the physiological mechanism of action by which these hormones govern body coloration is poorly understood. The protogynous wrasse Pseudolabrus sieboldi is a good model for understanding the physiological mechanisms of gonadal and body color change during sex change in hermaphrodite fish. To obtain information on the relationship between sex steroids and body color change during the process of gonadal sex change, we analyzed body color, gonadal histology, and serum levels of sex steroids. Body color was analyzed using a quantitative analytical method based on the hue value. Compared to other body parts of the fish, the anal fin changed color the most, becoming increasingly redder in association with gonadal changes that converted ovaries to testes. Levels of serum 11-ketotestosterone (11KT) increased as the gonadal sex change proceeded, whereas no significant change was observed in estradiol-17β (E2) levels. Moreover, we found a significant correlation between the hue value of the anal fin and serum 11KT levels, but not E2 levels. These results suggest that androgen, but not estrogen, plays a principle role in the changes in both gonadal morphology and body color in the transformation from female to male in this species. To our knowledge, this is the first quantitative demonstration of the relationship between body color and serum steroid levels during sex change in fish.     

Annual reproductive cycle of the captive female Japanese sardineSardinops melanostictus: Relationship to ovarian development and serum levels of gonadal steroid hormones

Gonad and blood samples were taken from the captive female Japanese sardineSardinops melanostictus between 1988 and 1989, and changes in serum levels of gonadal steroids were correlated with the annual gonadal cycle. Under captive conditions, female fish did not mature and spawn spontaneously, although oocytes developed up to the end of vitellogenic growth. Based on evidence from ovarian histology, the annual gonadal cycle of the Japanese sardine was divisible into four periods, i.e., immature (June to October), vitellogenesis (November to December), spawning (January to March), and post-spawning (April to May). The pattern of seasonal change in the gonadosomatic index (GSI) showed an inverse correlation to change in water temperature and reflected the degree of ovarian maturity. The serum estradiol-17 level increased from its lowest concentration (0.12 ng ml^–1) in September to a peak (1.14 ng ml^–1) in March. Serum 17,20-dihydroxy-4-pregnen-3-one (17,20-P) was detectable at low levels (<0.3 ng ml^–1) between October and February, but was below the assay detection limit (0.06 ng ml^–1) at all other times. Testosterone was not detectable (<0.06 ng ml^–1) in the serum of any fish throughout the year. The effects of several steroids on the maturation of follicle-enclosed oocytes of sardine were examined in vitro, and 17,20-P was found to be the most potent inducer of maturation. This suggests that post-vitellogenic oocytes of the Japanese sardine in captivity have an ability to respond to an appropriate hormonal effector and subsequently to resume meiotic maturation.     

Occurrence and detrimental effects of the bivalve-inhabiting hydroid <Emphasis Type="Italic">Eutima japonica</Emphasis> on juveniles of the Japanese scallop <Emphasis Type="Italic">Mizuhopecten yessoensis </Emphasis>in Funka Bay,Japan: relationship to juvenile massive mortality in 2003

In November 2003, we first observed prevalent occurrence of a hydroid, Eutima japonica, on soft body tissues of age zero Japanese scallop (Mizuhopecten yessoensis) juveniles cultured in large areas of Funka Bay, Hokkaido. The occurrence coincided with massive death of juvenile scallops. A major objective was to clarify ecological relationships between the symbionts, and to infer the relationship between symbiosis and the massive mortality. To do this, we investigated distributions of association rates of hydroids with juvenile scallops at 15–34 sites over 3 years (2003–2005), with age one adult scallops at 24 sites in 2003, and with mussels at 13 sites in 2004. We studied seasonal changes in association rates with juvenile scallops, and numbers of polyps per juvenile scallop at three sites from November 2003 to June 2004. We also quantified the hydroid impacts on juvenile scallop shell length growth and triglyceride accumulation in the digestive gland. The association rate of E. japonica polyps with juvenile scallops was high in large areas of Funka Bay in 2003, and overlapped the distribution of mussels bearing polyps. Association rates with age one adult scallops were very low in November 2003, even at the sites where polyps were very common on juvenile scallops. Levels of hydroid occurrence in juvenile scallops varies by year. We found that hydroids presence in juvenile scallops declined drastically in 2004 and 2005. The association rates with juvenile scallops, and numbers of polyps per juvenile scallop declined during winter, until they disappeared completely in the following June. Since polyps were rare in adult scallops, we believe that infection of juvenile scallops was probably initiated from the planulae produced by medusae released from polyps growing on Mytilus spp., especially M. galloprovincialis. Subsequently, the inhabitation spread intraspecifically and interspecifically. In juvenile scallops, inhabitation of polyps reduced shell length growth by 43%, and triglyceride accumulation in digestive glands by 24–47%. Inhabitation of E. japonica on juvenile scallop is best regarded as parasitism, rather than inquilinism or commensalism. Occurrence of polyps was probably not a direct lethal factor for juvenile scallops, because there were some sites where association rates were high, but mortalities were low. Massive mortalities in 2003 may have resulted from simultaneous impacts of heavy polyp load and stresses caused by the way in which the animals were handled (transferred from cages for pre-intermediate culture to cages for intermediate culture), because the massive mortality occurred within a month of the transfer. The presence of polyps in juvenile scallops does not affect the quality of the product in Funka Bay, because market size scallops are hydroid-free.     

A method for diagnosis of plant environmental stresses by gene expression profiling using a cDNA macroarray

Plants in the field are subjected to numerous environmental stresses. Lengthy continuation of such environmental stresses or a rapid increase in their intensity is harmful to vegetation. Assessments of the phytotoxicity of various stresses have been performed in many countries, although they have largely been based on estimates of leaf injury. We developed a novel method of detecting plant stresses that is more sensitive and specific than those previously available. This method is based on the detection of mRNA expression changes in 205 ozone-responsive Arabidopsis expressed sequence tags (ESTs) by cDNA macroarray analysis. By using this method, we illustrated shifts in gene expression in response to stressors such as drought, salinity, UV-B, low temperature, high temperature, and acid rain, as distinct from those in response to ozone. We also made a mini-scale macroarray with 12 ESTs for diagnosis of the above environmental stresses in plants. These results illustrate the potential of our cDNA macroarray for diagnosis of various stresses in plants.     

Enzymatic treatment of estrogens and estrogen glucuronide

Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers.In this article we investigated the enzymatic treatment of three estrogens (estrone,17β-estradiol,and 17α-ethynyletstradiol) by a fungal laccase which oxidize phenolic compounds with dissolved oxygen.The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay.In addition,we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-(β-D-glucuronide) degradation.The two enzymes eliminated 17β-estradiol 3-(β-D-glucuronide) and the intermediate,17β-estradiol,efficiently.     

Time-resolved measurements of aerosol elemental concentrations in indoor working environments

We have measured the elemental concentrations in aerosols with a 2-h time resolution in two different types of working environment: a chemistry laboratory dealing with the processing of advanced nanoparticulate materials and a medium-sized machine workshop. Non-stop 10-day and 12-day samplings were performed at each location in order to determine the concentration trends during the non-working/working and weekday/weekend periods. Supplementary measurements of PM10 aerosols with a 2-day sample collection time were performed with a standard Gent PM10 sampler to compare the elemental concentrations with the time-averaged concentrations detected by the 2D step-sampler. The concentrations were determined a posteriori by analyzing the x-ray spectra of aerosol samples emitted after 3-MeV proton bombardment. The PM10 samples collected in the chemistry laboratory were additionally inspected by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) to determine the chemical compositions of the individual particles. In the workshop, a total PM10 mass sampling was performed simultaneously with a minute resolution to compare the signal with typical outdoor PM10 concentration levels. A factor analysis of the time-resolved dataset points to six and eight factors in the chemistry laboratory and the machine workshop, respectively. These factors describe most of the data variance, and their composition in terms of different elements can be related to specific indoor activities and conditions. We were able to demonstrate that the elemental concentration sampling with hourly resolution is an excellent tool for studying the indoor air pollution. While sampling the total PM10 mass concentration with a minute resolution may lack the potential to identify the emission sources in a “noisy” environment, the time averaging on a day time scale is too coarse to cope with the working dynamics, even if elemental sensitivity is an option.