In vivo visual reporter system for detection of estrogen-like substances by transgenic medaka

Detection of endocrine disrupting chemicals, in particular, environmental estrogens with living organisms, has many advantages if compared to chemical analysis. The screening of novel pollutants with meaningful endpoints, the integration of uptake, bioconcentration, and excretion as well as the evaluation of endocrine disrupting effects with respect to toxicity require in vivo biotests for estrogen-like substances (ELSs). Critical disadvantages of whole organism biotests are their low sensitivity and the need for laborious and time-consuming work. To overcome these problems, we have developed a transgenic medaka strain harboring the green fluorescence protein (GFP) gene driven by choriogenin H gene regulatory elements. Choriogenin H is an egg envelope protein induced by estrogens in the liver. With yolk sac larvae of this strain, GFP induction in liver was observed 24 h after onset of aqueous exposure to 0.63 nM 17beta-estradiol (E2), 0.34 nM ethynylestradiol, or 14.8 nM estrone. Furthermore, concentrated sewage treatment effluent induced GFP expression. Comparison of E2 equivalents estimated by GFP-induction in transgenic medaka, a YES assay, and GC/MS showed detection limits in the same order of magnitude. These results indicated that the sensitivity of the transgenic medaka strain was sufficient for application as an alternative model in monitoring environmental water samples for ELSs.     

Homogeneous Tritylation of Cellulose in A Sulfur Dioxide – Diethylamine – Dimethyl Sulfoxide Medium

The present investigation was undertaken to see if a practical method could be developed for homogeneous tritylation of cellulose in a non-aqueous solvent of cellulose. Our new procedure of tritylation of cellulose can easily be carried out under homogeneous conditions by dissolving cellulose in a sulfur dioxide(SO₂)-diethylamine (DEA)-dimethyl sulfoxide(DMSO) solvent system, one or the non-aqueous cellulose solvents, followed by addition of trityl chloride and pyridine. This new method can avoid the time consuming pretreatment for the decrystallization of cellulose which has been necessary in the traditional procedure and can lower the reaction temperature. IR spectra of the products indicated the formation of trityl cellulose. Measurements of dielectric properties of the products confirmed that trityl groups were selectively introduced at the primary hydroxyl groups in cellulose. This conclusion was also confirmed by a H-NMR study in which the tritylated products was first acetylated, detritylated and then trideuterioacetylated and H-NMR spectra were taken at each stage and examined comparatively.     

The preparation,swelling characteristics,and albumin adsorption and release behaviors of a novel chitosan-based polyampholyte hydrogel

A novel chitosan (Ch)-based polyampholyte hydrogel was prepared from Ch dissolved in a 1:1 (v/v) mixture of 10% aqueous acetic acid and N-methyl-2-pyrrolidinone (NMP) by simple crosslinking using 1,2,3,4-butanetetracarboxylic dianhydride (BTCA). The detailed structure of the hydrogel was determined via FTIR and solid-state ¹³C NMR spectroscopic analyses. The swelling behavior of the hydrogels was strongly dependent on the BTCA feed ratio, and the hydrogels exhibited a pH-responsive swelling ratio that was influenced by the presence of both cationic NH₃⁺ and anionic COO^? groups within their molecular structures. The Ch hydrogels also exhibited bovine serum albumin (BSA) adsorption capacity, which was maximal at pH 4.5, consistent with the isoelectric point of BSA (4.7). In addition, the BSA adsorption capacity of the hydrogel decreased with the increasing ionic strength of the adsorption medium, indicating that the capacity of the hydrogel to adsorb BSA is facilitated by hydrophobic as well as electrostatic interactions between the hydrogels and the BSA molecules. In addition, a high desorption ratio (89%) of BSA was achieved in aqueous solutions at pH 2.0.     

Phase stability among the α(A1), β(A2), and γ(D83) phases in the Cu-Al-X system

The phase equilibria among the α(A1), β(A2), and γ(D8₃) phases in the Cu-Al-X systems (X: Ti, V, Mn, Fe, Co, Ni, Zn, Sn, and Sb) at 700 and 800 °C were investigated, and the effects of these alloying elements on the phase boundaries, homogeneous solid solution ranges, and tie-lines among the α, β, and γ phases were accurately determined by the diffusion couple method. The phase stabilities among α, β, and γ phases are discussed in terms of the partition coefficient. In addition, a modified Guillet’s method, which is used to estimate the effect of alloying elements on microstructure, is proposed by taking into account the change of equivalent coefficient with the phase fraction, and applied to predict the effect of alloying elements on the microstructure of the Cu-Al base alloy.     

Phase equilibria of Sn-In based micro-soldering alloys

The phase equilibria of Sn-In-X (X=Ag, Bi, Sb, Zn), the most basic information necessary for the development of Pb-free micro-soldering alloys, were studied using the CALPHAD method. Thermodynamic analyses for describing the Gibbs energies of the constituent phases were made by optimizing the obtained data on the experimental phase diagrams, and such data in the literature, including data on thermochemical properties. The present results combined with the thermodynamic database which was recently developed by our group [I. Ohnuma et al., J. Electron. Mater. 28, 1164 (1999)] provide various information on phase equilibria such as liquidus and solidus surfaces, isothermal and vertical section diagrams, mole fractions of the phase constitutions, etc., and thermodynamic properties such as activity, heat of mixing, surface energy, viscosity, etc., in multi-component soldering alloy systems including the elements of Pb, Bi, Sn, Sb, Cu, Ag, Zn, and In. Typical examples for the phase diagrams and thermodynamic properties of Sn-In-X ternary systems are shown. The application of the database to the alloy design for Pb-free solders is also presented.     

Effects of dietary defatted squid on cholesterol metabolism and hepatic lipogenesis in rats

Male Sprague-Dawley rats were fed a cholesterol-free (Exp. 1) or cholesterol-supplemented (Exp. 2) diet containing 20% casein (control group) or 15% defatted squid and 5% casein (defatted squid group), as protein, for 14 d. Serum and hepatic cholesterol concentrations were lower in rats fed defatted squid than in those fed casein in both cholesterol-free (−20%, P<0.05 and −15%, P<0.05, respectively) and cholesterol-supplemented (−25%, P<0.05 and −15%, P<0.05, respectively) diets. Hepatic triglyceride concentration was lower in the defatted squid than in the control groups in both cholesterol-free (−51%, P<0.05) and cholesterol-supplemented diets (−38%, P<0.01). The activities of cytosolic fatty acid synthase and the NADPH-generating enzymes, malic enzyme and glucose-6-phosphate dehydrogenase, in the liver were lower in the defatted squid than in the control groups in both cholesterol-free (−21%, P<0.01, −33%, P<0.05, and −33%, P<0.01, respectively) and cholesterol-supplemented diets (−34%, P<0.05, −57%, P<0.05, and −67%, P<0.05, respectively). The activity of mitochondrial carnitine palmitoyltransferase in the liver was comparable between the control and defatted squid groups. The activity of Mg²⁺-dependent phosphatidate phosphohydrolase in the liver cytosol was lower in the defatted squid (−9%, P<0.05) than in the control groups only in the cholesterol-free diet. Fecal excretion of total steroids was stimulated by the feeding of defatted squid in both cholesterol-free (+77%, P<0.005) and cholesterol-supplemented diets (+29%, P<0.01). These results suggest that the nonlipid fraction of squid exerts a hypocholesterolemic effect by increasing the excretion of total steroids in feces. The fraction also induces a triglyceride-lowering activity in the liver by decreasing hepatic lipogenesis.     

The Mucus Binding Factor Is Not Necessary for Lacticaseibacillus rhamnosus CRL1505 to Exert Its Immunomodulatory Activities in Local and Distal Mucosal Sites

Both viable and non-viable orally administered Lacticaseibacillus rhamnosus CRL1505 modulate immunity in local (intestine) and distal (respiratory) mucosal sites. So, intestinal adhesion and colonization are not necessary for this probiotic strain to exert its immunomodulatory effects. In this work, a mucus-binding factor knockout CRL1505 strain (ΔmbfCRL1505) was obtained and the lack of binding ability to both intestinal epithelial cells and mucin was demonstrated in vitro. In addition, two sets of in vivo experiments in 6-week-old Balb/c mice were performed to evaluate ΔmbfCRL1505 immunomodulatory activities. (A) Orally administered ΔmbfCRL1505 prior to intraperitoneal injection of the Toll-like receptor 3 (TLR3) agonist poly(I:C) significantly reduced intraepithelial lymphocytes (CD3⁺NK1.1⁺CD8αα⁺) and pro-inflammatory mediators (TNF-α, IL-6 and IL-15) in the intestinal mucosa. (B) Orally administered ΔmbfCRL1505 prior to nasal stimulation with poly(I:C) significantly decreased the levels of the biochemical markers of lung tissue damage. In addition, reduced recruitment of neutrophils and levels of pro-inflammatory mediators (TNF-α, IL-6 and IL-8) as well as increased IFN-β and IFN-γ in the respiratory mucosa were observed in ΔmbfCRL1505-treated mice when compared to untreated control mice. The immunological changes induced by the ΔmbfCRL1505 strain were not different from those observed for the wild-type CRL1505 strain. Although it is generally accepted that the expression of adhesion factors is necessary for immunobiotics to induce their beneficial effects, it was demonstrated here that the mbf protein is not required for L. rhamnosus CRL1505 to exert its immunomodulatory activities in local and distal mucosal sites. These results are a step forward towards understanding the mechanisms involved in the immunomodulatory capabilities of L. rhamnosus CRL1505.     

Performance improvement of the filter-type ‘transXend’ energy-resolving detector by considering noise sensitivity

The filter-type ‘transXend’ detector is an energy-resolving X-ray detector consisting of an energy-integrating flat-panel detector and multiple filters. In our previous studies, we have shown the effectiveness of the transXend detector, but the filters used were not optimized. To obtain better performance, the filters, especially their thickness and material, should be considered. In this paper, we present a method that can preliminarily estimate filter performance by comparing their noise sensitivity before carrying out numerous experiments. Two kinds of filter sets, Cu–Sn and W–Ag, with various thicknesses were evaluated. The results suggest that to image a 20-cm-thick object with 120-kV X-rays, an unfiltered channel and a channel filtered with 0.5-mm-thick Sn or 0.4-mm-thick Ag may be the best combination. The optimal filter thickness will be smaller if the object is thinner and the tube voltage is lower. For applications that require a wide dynamic range of detector response, a channel with a W filter may be better than an unfiltered channel. To verify the calculation results, computed tomography imaging experiments with a 3-cm-diameter phantom were also performed, and the experimental results showed good agreement with the calculation results.