A field study on serum vitellogenin levels in male Reeves' pond turtles (Chinemys reevesii) from estrogen-contaminated sites and a reference site

To ascertain whether wild male turtles were influenced by environmental estrogens, we examined serum vitellogenin (VTG) levels of male Reeves' pond turtles (Chinemys reevesii) collected from four study sites (A-D) in Kyoto, Japan. Sites A-C, which were impacted by domestic or industrial wastewater and effluents from sewage treatment plants, were chosen as contaminated sites, and site D was intended as a reference site. This contaminated/reference site characterization was confirmed by measuring estrogenic activities of the water samples collected at each site for over a year. Serum VTG levels in the turtles were quantified by an enzyme-linked immunosorbent assay established previously. Estrogenic activities of the water samples were measured using a previously validated yeast two-hybrid assay and expressed as the estradiol-17beta equivalent. Estrogenic activity was observed at high levels at sites A-C, but was almost undetected at site D throughout the sampling period: the mean and range were 0.74 (<0.07-2.1), 0.52 (0.17-1.6), 1.7 (<0.07-7.3), and 0.07 (<0.07-0.62) ng/l at sites A-D, respectively. Significant differences were found only in site D versus sites A, B, and C. Therefore, site D and sites A-C were confirmed to be a reference site and contaminated ones, respectively. Overall, 320 male turtles were captured and examined. The majority of the turtles showed normal VTG values (0.10-0.74 microg/ml). Although only five turtles from sites A-C showed unusually high VTG values (1.1-5.9 microg/ml, nearly one order of magnitude higher than normal values but much lower than values in adult females), there was no significant difference in the incidence of these high values between sites A-C and site D. Moreover, among the five turtles, one turtle was captured again 2 months later, but its VTG value dropped to the normal level. The unusually high VTG values may therefore be transient elevation caused by incidental and/or individually specific agents. Excluding the unusually high values, the mean serum VTG (accompanied with the range) was 0.22 (0.10-0.74), 0.27 (0.11-0.62), 0.27 (0.17-0.68), and 0.23 (0.10-0.57) microg/ml at sites A-D, respectively. There were no significant differences in the mean VTG values among sites A-D. Although it should be clarified how and why the unusually high VTG values occurred at sites A-C, our results suggested that wild male C. reevesii turtles would not be significantly affected by xenobiotic estrogens at environmentally relevant levels in terms of serum VTG elevation.     

Relationships between Bacteroides 16S rRNA genetic markers and presence of bacterial enteric pathogens and conventional fecal indicators

Occurrence and prevalence of different bacterial enteric pathogens as well as their relationships with conventional (total and fecal coliforms) and alternative fecal indicators (host-specific Bacteroides 16S rRNA genetic markers) were investigated for various water samples taken from different sites with different degrees of fecal contamination. The results showed that a wide range of bacterial pathogens could be detected in both municipal wastewater treatment plant samples and in surface water samples. Logistic regression analysis revealed that total and human-specific Bacteroides 16S rRNA genetic markers showed significant predictive values for the presence of Escheriachia coli O-157, Salmonella, heat-labile enterotoxin (LT) of enterotoxigenic E. coli (ETEC), and heat-stable enterotoxin for human (STh) of ETEC. The probability of occurrence of these pathogenic bacteria became significantly high when the concentrations of human-specific and total Bacteroides 16S rRNA genetic markers exceeded 10(3) and 10(4) copies/100 mL. In contrast, Clostridium perfringens was detected at high frequency regardless of sampling sites and levels of Bacteroides 16S rRNA genetic markers. No genes related to Shigella spp., Staphylococcus aureus and Vibrio cholerae were detected in any samples analyzed in this study. Conventional indicator microorganisms had low levels of correlation with the presence of pathogens as compared with the alternative fecal indicators. These results suggested that real-time PCR-based measurement of alternative Bacteroides 16S rRNA genetic markers was a rapid and sensitive tool to identify host-specific fecal pollution and probably associated bacterial pathogens. However, since one fecal indicator might not represent the relative abundance of all pathogenic bacteria, viruses and protozoa, combined application of alternative indicators with conventional ones could provide more comprehensive pictures of fecal contamination, its source and association with pathogenic microorganisms.     

Effect of operating factors on liquid–liquid mass transfer and dispersion pattern of sedimentary liquid in a mechanically stirred vessel

Liquid–liquid dispersion and mass transfer were investigated in mechanically stirred vessels without baffles by changing operation factors such as an impeller rotation speed, off-bottom clearance, volumetric liquid ratio, etc. The dispersion regime was categorized into five groups: the sedimentary liquid was kept at the vessel bottom (I), partially elevated without any collision (II), partially dispersed by colliding with the impeller bottom (III), both liquids were partially dispersed by collisions with impeller blades (III’), and the sedimentary liquid was completely dispersed (IV). The dispersion switched to I → II → III → IV with the increasing rotation speed and decreasing off-bottom clearance. The liquid–liquid mass transfer rate was significantly enhanced with the collision of the sedimentary liquid with the impeller bottom, and subsequently increased with the increasing rotation speed, volumetric liquid ratio, and vessel diameter and with the decreasing off-bottom clearance. A multiple regression analysis method was applied to determine the mass transfer rates of III and III’.     

CONTENTS OF CYTOCHROMES IN YEAST

The contents of cytochromes in yeast were determined quantitatively from the absorption spectra, using a solid cell paste of intact yeast. During the industrial production of baker's yeast, the contents of the cytochromes, particularly of cytochrome aa₃ at successive stages, increased gradually with increasing aeration. In semi-aerobically grown baker's yeast, the contents of cytochromes aa₃, b and c were 0·9, 2·9 and 2·9 × 10^?5 moles/litre of fresh yeast (total amount 6·7 × 10^?5 moles/litre), while in vigorously aerated commercial baker's yeast the respective values were 2·3, 4·8 and 5·2 × 10^?5 moles/litre (total amount 12·3 × 10^?5 moles/litre). In brewer's yeasts separated after the brewing process, the contents of cytochromes were markedly lower than in baker's yeast grown with limited aeration, whereas in top-fermenting yeast the total cytochrome content, aa₃ + b + c, was in some samples markedly higher, 7·1 × 10^?5 moles/litre, than in bottom-fermenting brewer's yeast, 2·4 × 10^?5 moles/litre. When brewer's bottom yeast was grown on a laboratory scale under increasing aeration, a maximum appeared in the cytochrome contents when aeration was moderate, and increased aeration inhibited the formation of cytochromes. The cytochrome contents in brewer's bottom yeast may exceed the amounts found in commercial baker's yeast. In addition to aeration, the type of metabolism influences the amounts of cytochromes in yeast.     

FREE NUCLEOTIDES IN RESTING AND METABOLIZING BAKER'S YEAST*

The free nucleotides of semi-aerobically and aerobically cultured baker's yeast in different industrial stages have been investigated under different conditions: during growth and at rest, and during respiration and fermentation of different substrates. The amounts of protein and nucleic acids diminish progressively during industrial propagation, while the amount of free nucleotides falls only during the last stage of propagation. In yeasts cultured under different conditions of aeration and nutrient addition, the nucleotide spectra were rather similar, but the proportions of the free nucleotides change considerably when the yeast passes from the resting state to growth or metabolism. On transfer from respiration to fermentation the nucleotide spectrum remained practically unchanged. However, a small addition of ethanol to a respiring yeast suspension caused clear alterations in the spectrum, the most obvious being the reduced amount of nucleotides with energy-rich phosphate bonds.     

Characterization of endogenous enzymes of milled rice and its application to rice cooking

The relationship between the activities of endogenous enzymes in milled rice and accumulation of chemical components in the rice grains during cooking was investigated. Maltose and soluble starch were optimally hydrolyzed around 60 °C by both crude extracts and purified α-glucosidases of milled rice. Gelatinization onset temperature of rice flour was determined to be 63 °C with DSC analysis under usual cooking condition. Hydrolytic activities on carbohydrates and proteolytic activities of milled rice were enhanced at pH 5 compared to pH 7 at the range of 4–80 °C. When rice was cooked at pH 5, glucose and amino acids highly accumulated with soaking for 16 h before heating. We propose a new method of cooking with prolonged soaking at acidic pH, which is effective for increasing the amounts of chemical components in cooked rice.     

Powdered formulation of liquid oil terpenoid included in cyclodextrins for chemical stabilization and improved handling

Teprenone (Tp) is a widely used pharmaceutically active ingredient in terpene anti-ulcer agents. We investigated Tp and cyclodextrin (CD) complexes for the objective of forming stable Tp powders. Tp was combined with each CD at a molar ratio of 1:2, kneaded together while adding an appropriate amount of water, then dried under reduced pressure and lightly milled into a powder to produce a kneaded mixture (KM). Powder X-ray diffraction of KM containing Tp and β- and γ-CD revealed new diffraction peaks. On infrared spectroscopy, the peak near 1718 cm^?1 corresponding to the carbonyl group of Tp had shifted to around 1701 cm^?1. Solid-state NMR spectroscopy showed signs of inhibited molecular mobility of Tp, which is liquid at room temperature, confirming that Tp had formed complexes with β- and γ-CD. Photostability analysis of Tp showed improved stability in the order of γ-CD > β-CD ? α-CD.     

Structural changes in carbon aerogels with high temperature treatment

The structural change of carbon aerogels at high temperatures up to 2800°C has been investigated. Change in microtexture of fine particles, which constitute carbon aerogels derived from phenolic resin, was of a typical non-graphitized carbon. The microporosity decreased with an increase of heat-treatment temperature, and disappeared at 2000°C. The mesoporosity still remained even after heat-treatment up to 2800°C, though 50% of mesopore volume was lost because of the fusion of the particles with the change of carbon microtexture.     

Modelling and prediction of phyto- and zooplankton dynamics in Lake Kasumigaura by artificial neural networks

An artificial neural network model was developed for Lake Kasumigaura to predict timing and magnitudes for chlorophyll a, five species of blue-green algae and three zooplankton groups. The model was trained by 8 years of limnological time series and validated by two independent years. The validation showed the potential of neural networks as predictive tools for highly non-linear phenomena such as blue-green algal blooms in freshwater lakes.