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The ratio of CO2 efflux to O2 influx (ARQ, apparent respiratory quotient) in tree stems is expected to be 1.0 for carbohydrates, the main substrate supporting stem respiration. In previous studies of stem fluxes, ARQ values below 1.0 were observed and hypothesized to indicate retention of respired carbon within the stem. Here, we demonstrate that stem ARQ < 1.0 values are common across 85 tropical, temperate, and Mediterranean forest trees from nine different species. Mean ARQ values per species per site ranged from 0.39 to 0.78, with an overall mean of 0.59. Assuming that O2 uptake provides a measure of in situ stem respiration (due to the low solubility of O2), the overall mean indicates that on average 41 % of CO2 respired in stems is not emitted from the local stem surface. The instantaneous ARQ did not vary with sap flow. ARQ values of incubated stem cores were similar to those measured in stem chambers on intact trees. We therefore conclude that dissolution of CO2 in the xylem sap and transport away from the site of respiration cannot explain the low ARQ values. We suggest refixation of respired CO2 in biosynthesis reactions as possible mechanism for low ARQ values.

Estimations of ecosystem-level evapotranspiration (ET) and CO2 uptake in water-limited environments are scarce and scaling up ground-level measurements is not straightforward. A biophysical approach using remote sensing (RS) and meteorological data (RS–Met) is adjusted to extreme high-energy water-limited Mediterranean ecosystems that suffer from continuous stress conditions to provide daily estimations of ET and CO2 uptake (measured as gross primary production, GPP) at a spatial resolution of 250 m. The RS–Met was adjusted using a seasonal water deficit factor (fWD) based on daily rainfall, temperature and radiation data. We validated our adjusted RS–Met with eddy covariance flux measurements using a newly developed mobile lab system and the single active FLUXNET station operating in this region (Yatir pine forest station) at a total of seven forest and non-forest sites across a climatic transect in Israel (280–770 mm yr−1). RS–Met was also compared to the satellite-borne MODIS-based ET and GPP products (MOD16 and MOD17, respectively) at these sites.Results show that the inclusion of the fWD significantly improved the model, with R = 0.64–0.91 for the ET-adjusted model (compared to 0.05–0.80 for the unadjusted model) and R = 0.72–0.92 for the adjusted GPP model (compared to R = 0.56–0.90 of the non-adjusted model). The RS–Met (with the fWD) successfully tracked observed changes in ET and GPP between dry and wet seasons across the sites. ET and GPP estimates from the adjusted RS–Met also agreed well with eddy covariance estimates on an annual timescale at the FLUXNET station of Yatir (266 ± 61 vs. 257 ± 58 mm yr−1 and 765 ± 112 vs. 748 ± 124 gC m−2 yr−1 for ET and GPP, respectively). Comparison with MODIS products showed consistently lower estimates from the MODIS-based models, particularly at the forest sites. Using the adjusted RS–Met, we show that afforestation significantly increased the water use efficiency (the ratio of carbon uptake to ET) in this region, with the positive effect decreasing when moving from dry to more humid environments, strengthening the importance of drylands afforestation. This simple yet robust biophysical approach shows promise for reliable ecosystem-level estimations of ET and CO2 uptake in extreme high-energy water-limited environments.

Obesity is a metabolic disorder associated with many diseases. WW domain-containing E3 ubiquitin protein ligase 1 (WWP1) is a HECT-type E3 ubiquitin ligase involved in several diseases. Recently, we found that the level of WWP1 is increased in white adipose tissue in a mouse model of obesity and that obese Wwp1 knockout (KO) mice exhibit improved whole-body glucose metabolism. Here, to determine which insulin-sensitive tissues contribute to this phenotype, we investigated the levels of several insulin signaling markers in white adipose tissue, liver, and skeletal muscle of Wwp1 KO mice, which were fed a normal or high-fat diet and transiently treated with insulin. In obese Wwp1 KO mice, phosphorylated Akt levels were increased in the liver but not in white adipose tissue or skeletal muscle. Moreover, the weight and triglyceride content of the liver of obese Wwp1 KO mice were decreased. These results suggest that systemic deletion of WWP1 improves glucose metabolism via enhanced hepatic insulin signaling and suppressed hepatic fat accumulation. In summary, WWP1 participates in obesity-related metabolic dysfunction and pathologies related to hepatic steatosis via suppressed insulin signaling., (© 2023 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

The scanning electron microscope (SEM) has been reassembled into a new type of cryo-electron microscope (cryo-TSEM) by installing a new cryo-transfer holder and anti-contamination trap, which allowed simultaneous acquisition of both transmission images (STEM images) and surface images (SEM images) in the frozen state. The ultimate temperatures of the holder and the trap reached - 190 °C and - 210 °C, respectively, by applying a liquid nitrogen slush. The STEM images at 30 kV were comparable to, or superior to, the images acquired with conventional transmission electron microscope (100 kV TEM) in contrast and sharpness. The unroofing method was used to observe membrane cytoskeletons instead of the frozen section and the FIB methods. Deep sublimation of ice surrounding unroofed cells by regulating temperature enabled to emerge intracellular fine structures in thick frozen cells. Hence, fine structures in the vicinity of the cell membrane such as the cytoskeleton, polyribosome chains and endoplasmic reticulum (ER) became visible. The ER was distributed as a wide, flat structure beneath the cell membrane, forming a large spatial network with tubular ER., (© 2021. The Author(s).)

Temporal changes in carrier relaxations, magnetic switching, and biological structures are known to be in the order of ns. These phenomena can be typically measured by means of an optical-pump & electron-probe method using an electron microscope combined with a pulsed electron source. A photoemission-type pulsed electron gun makes it possible to obtain a short-pulsed electron beam required for high temporal resolution. On the other hand, spatial resolution is restricted by the brightness of the pulsed electron gun used in electron microscopes when a low brightness electron source is used and an irradiation current larger than a certain value is required. Thus, we constructed a prototype pulsed electron gun using a negative electron affinity (NEA) photocathode for time-resolved measurement using a scanning electron microscope (SEM) with high spatiotemporal resolution. In this study, a high-speed detector containing an avalanche photodiode (APD) was used to directly measure waveforms of the pulsed electron beam excited by a rectangular-shape pulsed light with a variable pulse duration in the range of several ns to several μs. The measured waveforms were the same rectangular shape as incident pulsed excitation light. The maximum peak brightness of the pulsed electron beam was 4.2×10 7 A/m 2 /sr/V with a pulse duration of 3 ns. This value was larger than that of the continuous electron beam (1.6 × 10 7 A/m 2 /sr/V). Furthermore, an SEM image with image sharpness of 6.2 nm was obtained using an SEM equipped with a prototype pulsed electron gun at an acceleration voltage of 3 kV., (Copyright © 2021. Published by Elsevier B.V.)

Eight structural components of humic substances were ozonated. Mutagenic activity was found using TA100 with and without S9 mix for all ozonated components. p-Hydroxybenzaldehyde was chosen as an important component and ozonation products were determined by gas chromatography-mass spectrometry (GC-MS). Aldehydes, ketones and carboxylic acids were identified as the ozonation products. Among these products, acetaldehyde, formaldehyde, glyoxal, methylglyoxal and glyoxylic acid were recognized to be mutagenic. Furthermore, p-hydroxybenzaldehyde was first ozonated and then chlorinated. A great variety of chlorinated organic compounds, many of which are known mutagens, have been identified by GC-MS in the ether extract. The same compounds have previously been reported as chlorination products of humic substances. Aldehydic products by ozonation were identified from ozonation followed by chlorination of humic substances and p-hydroxybenzaldehyde.

Components of humic substances, such as vanillin, syringaldehyde, vanillic acid and di-n-butylphtalate, were ozonated and subjected to the mutagenicity assay using Salmonella typhimurium TA 98 and 100 with and without S9 mix. The strong mutagenic activity was found on all components except di-n-butylphtalate by strain TA 100 with and without S9 mix. Substances with strong mutagenic activity in ozonated vanillin were water-soluble and were slightly extracted with benzene, dichloromethane and ethyl acetate. Following gel chromatography on Sephadex G-10, the strong mutagens generated by ozonation were found with molecular weights greater than 300.

Humic substances and p-hydroxybenzaldehyde, one of their components, were ozonated and quantitative analysis of the mutagenic aldehydes (formaldehyde, acetaldehyde, glyoxal, glyoxylic acid and methylglyoxal) was performed. Glyoxal and glyoxylic acid were the main mutagenic compounds. The ozone-treated solutions were flowed through a granular activated carbon (GAC) column and the KMnO4 consumed of the effluent decreased to about 40-50%. Most of the aldehydes formed by ozonation reduced, but glyoxal increased.

The modulating effects of the Chinese medicinal plant 'Tan-shen', the radix of Salvia miltiorrhiza Bunge, on the mutagenic activities of Trp-P-1 (3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole) and B(a)P (benzo[a]pyrene) were investigated using Salmonella typhimurium TA98. Ether- and hot water-extracted 'Tan-shen' enhanced both mutagens at low concentrations, but suppressed them at high concentrations. Extracts by ether treatment were more effective than those extracted by hot water. Dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone IIA were isolated from the ether extract by high performance liquid chromatography (HPLC) and were recognized to be the mutagenic modulators. 4 tanshinones enhanced the mutagenicity of Trp-P-1 by 8-24-fold at 20 micrograms/plate and the enhancement was reduced at the higher concentration. Dihydrotanshinone I suppressed Trp-P-1 activity completely at 100 micrograms/plate.

p-Hydroxybenzaldehyde, a component of soil humic substances, was ozonated and chlorinated. The ether extract and the residue were subjected to the Ames assay; mutagenic activities were identified. The non-ionic resin CSP800 and the anion exchange resin CHPA25 were used for separation of mutagenic compounds. The compounds in the water layer were not adsorbed on CSP800 or CHPA25 and exhibited strong mutagenic activity. Mutagenic activity was reduced as the added chlorine was increased. Ether extracts were analyzed by gas chromatography-mass spectrometry (GC-MS) and chloral, 1,3-dichloro-2-propanone, 1,2,3-trichloro-1-propene, tetrachloroethylene and 1,1,1,3,3-pentachloro-2-propanone were identified as mutagenic compounds.

Eight components of humic substances were ozonated. Mutagenic activity was found with TA100 with and without S9 mix for all ozonated components. Ozonated products of p-hydroxybenzaldehyde were separated into five fractions by silica gel chromatography and each fraction subjected to mutagenicity assay. Mutagenic activity was found in the chloroform and chloroform-acetone (1:1) fractions. The compounds in these fractions were identified, and aldehydes such as formaldehyde, acetaldehyde, glyoxal, glyoxylic acid and methyl glyoxal were found to be mutagenic. Mutagenic compounds are present in the polar fraction.

The SOS chromotest was applied for the detection of antimutagens. To raise SOS induction, the bacteria were treated with the mutagens, UV, 4-nitroquinoline N-oxide (4NQO), N-methyl-N'-nitro-N-nitroso-guanidine (MNNG), or benzo[a]pyrene (B[a]p). The inhibitory effects of L-ascorbic acid, glutathione, vanillin, 5-fluorouracil (5-FU), 5-chlorouracil (5-CU), cobaltous chloride, sodium selenite and sodium arsenite, which are known as antimutagens, were investigated with their addition either simultaneously or post treatment time. It became clear that the SOS chromotest was very useful for the detection of antimutagens.

The mechanism of antimutagenicity of water extracts of grass-wrack pondweed (Potamogeton oxyphylus Miquel), curled pondweed (Potamogeton crispus L.) and smartweed (Polygonum hydropiper L.) towards benzo[a]pyrene mutagenicity in Salmonella typhimurium was investigated. The antimutagenic components in the aquatic plants were water-soluble, heat-resistant and had a high molecular weight; chlorophyll did not play an important role.

Boiling-water extract of Korean-Saiko (Bupleuri Radix, from South Korea, Bupleurum falcatum L.) enhanced the mutagenic activity of Trp-P-1, Trp-P-2 and benzo[a]pyrene with S9mix. The boiling-water extract was fractionated with ether and then n-BuOH. Both the ether and the n-BuOH fractions also enhanced mutagenicity of Trp-P-1, respectively. The n-BuOH fraction was separated into seven fractions by silica gel chromatography and the chloroform eluate had the strongest enhancing effect on the mutagenic activity of Trp-P-1 with S9mix. The chloroform eluate fraction was further separated into five spots by thin-layer chromatography. Two of the spots had the strongest enhancing effect on the mutagenic activity of Trp-P-1. Since saikosaponin is a well known component in Bupleuri Radix, the effects of its existence were tested and saikosaponins a and c were found. The enhancement activity of saikosaponin a was very weak. The effective components are now being studied.

The inhibitory effects of peony root extract on the mutagenicity of benzo[a]pyrene (B [a]p) have been investigated in the Salmonella typhimurium reversion test. Four kinds of experiments were performed: direct chemical reaction (1) between peony root extract and B [a]p, and (2) between peony root extract and active metabolite(s) of B [a]p, (3) inhibition of metabolic processes of B[a]p with S9 mix, and (4) inhibition of activation on mutagenicity. Peony root extract interfered with the action of enzymes in the S9 mix, and inactivated the activity of B[a]p metabolites. The bio-antimutagenic effect was assayed by reversion in Salmonella typhimurium TA98 and TA100.

Biological tests using Orizias latipes (LC50 and oxygen uptake test), Moina macrocopa (LC50), and Dugesia japonica (head regeneration test and LC50) were carried out in order to clarify the mutual relationship of these test methods. The oxygen uptake rate of O. latipes was not effective to assess chemical toxicity. Adding the results of the growth inhibition test of Tetrahymena pyriformis (Yoshioka, Y., Ose, Y., and Sato, T. (1985). Sci. Total Environ. 43, 149-157), the correlation coefficients between each two test methods were calculated. The test results except EC50 and LC50 of D. japonica showed a good relation to each other. We determined the solubility and the n-octanol/water partition coefficient (P) of some chemicals used in the test. Log P interpreted the toxicity in mol/liter unit but not in mg/liter. Solubility was not a useful descripter neither in mol/liter nor in mg/liter unit.

The effects of medicinal plants on the mutagenicity of benzo[a]pyrene were studied with Salmonella typhimurium tester strains. The chosen medicinal plants are very frequently used as Chinese herbal medicines. Each medicinal plant was extracted with hot water, which is similar to the method used in Chinese medicinal treatment. Cinnamomi cortex, Rhei rhizoma, Scutellariae radix and Rehmanniae radix were found to decrease the mutagenic activity of benzo[a]pyrene. Atractylodis rhizoma also reduced the mutagenicity of benzo[a]pyrene, but this was not certain, because it showed a killing effect on the cell survival test. Bupleuri radix and Aurantii nobilis pericarpium had an enhancing effect, but then neither of these extracts is itself mutagenic. Each medicinal plant extract showed a different effect on the mutagenicity of benzo[a]pyrene. These effects were classified into 5 types: (I) decreasing effect, (II) killing effect, (III) enhancing effect, (IV) enhancing and decreasing effect and (V) inactive.

Forty preparations of the extracts from 28 kinds of Asian herbs were tested for ability to inhibit the activities of murine retroviral reverse transcriptase and human deoxyribonucleic acid (DNA) polymerases. Among the 40 extracts, 35 inhibited reverse transcriptase activity and 29 inhibited DNA polymerase alpha activity. The inhibitory potencies of these extracts were expressed as the 50% inhibition concentrations (IC50), at which the enzyme activities were inhibited by 50%. Very strong inhibitions were observed with the extracts from Millettia pachycarpa (Leguminosae) and Mallotus apelta (Euphorbiaceae) as shown by their low IC50 values for reverse transcriptase (0.4-0.5 micrograms/ml) and DNA polymerase alpha (0.9-1.4 micrograms/ml). Enzyme kinetic analysis revealed that the mode of inhibition of reverse transcriptase by these two extracts was competitive with respect to the template.primer [poly(rA).oligo(dT)] and noncompetitive with respect to deoxythymidine triphosphate (dTTP) substrate. Besides reverse transcriptase and DNA polymerase alpha, DNA polymerase I and ribonucleic acid (RNA) polymerase from E. coli were inhibited by these two extracts. These results indicate that the herb extracts contain as yet unidentified substance(s) which inhibit the activities of reverse transcriptase and cellular DNA polymerases.

The water solubilities (S), octanol/water partition coefficients (KOW) and bioconcentration factors (BCF) of four polydimethylsiloxane (PDMS) fluids covering a wide range of molecular weight were measured. It is shown that a previously described correlation between S and KOW for organic chemicals may be invalid for PDMS fluids; an alternative correlation is proposed. Some PDMS fluids tend to have a bioconcentration potential in silver carp.

When carp (Cyprinus carpio) were exposed to 5 and 30 ppm Cd in the water, the contents of Cd-binding protein, which has low molecular weight, increased in the hepatopancreas, kidney, gills and gastrointestinal tract with the duration of exposure. This Cd-binding protein was purified from hepatopancreas, kidney, gills, and spleen of carp administered 2 mg/kg Cd (as CdCl2), intraperitoneally for 6 days. Two Cd-binding proteins were separated by DEAE-Sephadex A-25 column chromatography. These proteins had Cd-mercaptide bond, high cysteine contents (ca. 29-34%), but no aromatic amino acids or histidine. From these characteristics the Cd-binding proteins were identified as metallothionein. By using antiserum obtained from a rabbit to which carp hepatopancreas MT-II had been administered, immunological characteristics between hepatopancreas MT-I, II and kidney MT-II were studied, and a slight difference in antigenic determinant was observed among them. By immunological staining techniques with horseradish peroxidase, the localization of metallothionein was investigated. In the nontreated group, metallothionein was present in the acinar cells of hepatopancreas and renal convoluted tubules. In the Cd-treated group (2 mg/kg IP daily for 3 days), metallothionein was present in the nuclei, sinusoids, and extracellular space of hepatopancreas, in addition to the acinar cells. Carp were bred in 1 ppm Cd, 5 ppm Zn solution, and tap water for 14 days, following transfer to 15 ppm Cd solution, respectively. The survival ratio was the highest in the Zn group followed by Cd-treated and control groups. The metallothionein contents increased in hepatopancreas and kidney in the order: Zn greater than Cd greater than control group.

Soil humic substances were chlorinated in solution, extracted with ether and subjected to mutagenicity assay. Mutagenicity was detected using Salmonella typhimurium TA 100, with or without S9 mix; mutagenic activity was found to be less with added S9 mix. Ether extracts were analyzed by gas chromatography-mass spectrometry (GC-MS) and various chlorinated and non-chlorinated compounds were detected, e.g. chlorinated esters, acetones and carboxylic acids, as well as non-chlorinated aromatics. The mutagenicity of the major chlorination products was examined. Chloral and 1,1,1,3,3-pentachloro-2-propanone (pentachloroacetone) were found to be mutagenic, and 1,1-dichloro-2-propanone (1,1-dichloroacetone) was a possible mutagen. The yields of these mutagens increased with an increase in chlorine concentration.

The antimutagenicity of extracts from crude drugs was studied by the Ames bioassay system. The crude drugs chosen were medical plants used very frequently as Chinese medicines. Each crude drug was extracted with hot water similar to the method of Chinese medical treatment. Antimutagenicity of the extract was found with 4 kinds of crude drugs, Paeoniae radix, Bupleuri radix, Hoelen and Glycyrrhizae radix. Each extract of the crude drug showed a different type of antimutagenic action from the others.

In a previous paper, direct-acting mutagenicity was reported in emission gas from incomplete municipal incineration using Salmonella typhimurium TA-98 and TA-100. This paper reports the detection of dinitropyrene (DNP) as a direct-acting mutagen using nitrogen selective gas chromatography. The gas-phase photochemical reaction of pyrene with nitrogen dioxide was examined in a quartz vessel with various reaction times and temperatures. 1-Nitropyrene (1-NP) was readily formed from pyrene in the absence of light irradiation and at low temperature, but DNP was not formed under similar conditions. DNP formation was catalyzed by nitric acid. The formation of DNP is dependent on light irradiation, temperature and HNO3 as catalyst. In a combustion source these factors affect the formation of DNP.

The mutagenicity of refuse leachate from a municipal incinerator was studied by liquid rec-assay and Ames assay. Volatile components were found to be negative, and nonvolatile components positive, in the Ames assay and the leachate was found to have DNA-damaging capacity in the liquid rec-assay with S-9 mix. PAHs derived from tobacco ash and carbonyl compounds generated by the putrefaction of foods were confirmed to be main contributors to DNA-damaging capacity and mutagenicity in refuse leachate.

An investigation into the adsorption and desorption of polydimethylsiloxane, PCBs, cadmium nitrate, copper sulfate, nickel nitrate and zinc nitrate by river sediments was carried out using either a flow-through system or a semi-static system. The material balance in the sediment compartment could be explained by the equation, dCs/dT= K1Cw-K2Cs. The adsorption rate constants (K1), desorption rate constants (K2) and concentration factors (K1/K2) were calculated. For hydrophobic chemicals, the K1's were independent of water solubility, but the K2's were relatively related to water solubility. For both hydrophobic chemicals and heavy metals, the concentration factors per fraction organic carbon were similar to the soil sorption coefficients (Koc), expressed on a organic carbon basis.

We have investigated the mutagenic activity of extracts from the wastewater of sewage treatment plants in municipal waste incinerators and evaluated the relative contribution of various routes of emission from the incinerator to the total output of mutagens. The mutagenicity of wastewater extracts from a complete combustion incinerator was 10% of that from an incomplete combustion unit. About 90% of all the mutagens produced in a municipal incinerator are discharged into the atmosphere as emission gases, and 10% are disposed of in the wastewater treatment plants. Most of the mutagens in wastewater treatment plants are not decomposed by normal aeration times, but are removed by adsorption onto suspended solids.

The genotoxic potential of 6 disinfectants and their 9 metabolites was investigated by umu test. In the tested disinfectants, glutaraldehyde showed positive genotoxicity independent of metabolic activation system and acrinol was positive only in the presence of S9 mixture. Alkylaminoethylglycine, benzalkonium chloride, chlorhexidine digluconate and methylrosaniline chloride were negative in the presence or absence of S9 mixture. In some metabolites of benzalkonium chloride, chlorhexidine digluconate or glutaraldehyde, only pyrogallol showed positive genotoxicity in the absence of S9 mixture and the activity was not affected by the metabolic activation system. Aniline, p-chloroacetoanilide, p-chloroaniline, p-chlorophenol, decabutyldimethylamine, glutaric acid, phenol and pyrocatechol did not induce umu gene expression independently of the presence of S9 mixture. The results in the umu test of these compounds were compared with their findings in the liquid rec-assay and Ames test. The umu test is a more useful and simplified method for the detection of genotoxicity of the compounds with killing effects on tester bacteria.

Twenty kinds of coal-tar dyes were subjected to median tolerance limit (TLm) test by use of Himedaka (Oryzias latipes) for the comparision of their acute toxicities. It became clear that 4 kinds of halogens substituted xanthene compounds dyes showed strong acute toxicities. From the fact that uranine had the lower acute toxicity than halogens substituted compounds and the toxicities of these 4 dyes increased through irradiation, it was assumed that halogen atoms in dyes might be responsible for these strong acute toxicities to fish.

Mutagenic activity was observed in sediments of the Suimon River bed with and without S9 mix. The direct-acting mutagens in the sediment were investigated. The sediment was extracted with methanol and fractionated on a Silica gel column. The benzene fraction from the Silica gel column exhibited mutagenic activity without S9 mix in strain TA98, while it failed to show mutagenic activity in nitroreductase-deficient strain TA98NR. This observation led to the suspicion that nitro compounds were the direct-acting mutagens of these samples. The benzene fraction was treated by heptafluorobutyric anhydride (HFBA) and investigated with gas chromatography equipped with an electron capture detector (GC-ECD). 2-Nitrofluorene, 4,4'-dinitrobiphenyl, 2,7-dinitrofluorene and 1-nitropyrene were detected and measured quantitatively. The mutagenic activity of a mixture of these compounds was compared with that of the original fraction and the direct-acting mutagenicity of Suimon River sediment can be explained by these nitroarenes, especially 1-nitropyrene.

Humic acid inhibited the mutagenicity of various mutagens. The inhibitory effect was desmutagenic, heat-resistant and increased with an increase of the humic acid molecular weight. Typical monomeric components of humic acids had no desmutagenic effect. The desmutagenic effect of humic acid was caused by adsorption of mutagen, not by decomposition of mutagen. The adsorption activity was largest at its critical micelle concentration and the adsorbed mutagen was released by ultrasonication. Humic acids exist in natural environment in large amounts and may play an important role for natural purification by adsorption of mutagens.

Reason of acute toxicity to fish caused by coal-tar dyes and their industrial effluents were studied from the three points of view. 1) Partition coefficient (n-octanol/water) was determined on several coal-tar dyes, and it was found that the larger the ratio was, the stronger was toxicity to fish. 2) As a result of determination about existent form of xanthene dyes under photo-irradiation, it was clarified that dyes in solution were in the form of excited triplet state and halogen in dye molecule was liberated in free form. 3) When coal-tar dyes were treated with sodium hypochlorite, harmful chloroform was detected from dye solution. Chloroform and its homologous compounds showed the low TLm value and had the serious effect on fish.

The DNA-damaging capacity and the mutagenicity of 6 disinfectants were studied by liquid rec-assay and Ames test. 5 disinfectants were found to be positive in DNA-damaging capacity while only one of them showed clear mutagenicity in the Ames test. Liquid rec-assay by direct incubation with S9 mix was the most sensitive method and gave the best correlation between the growth ratio (R 50) and the time lag, both of which compared Rec+ and Rec-. Liquid rec-assay may be useful for detecting the DNA-damaging capacity of chemicals with a strong killing effect.

A new analytical method for assaying polyorganosiloxanes (silicones) in environmental samples is presented. The method utilizes petroleum for solvent extraction of the sample (water, sediment, biological tissue) together with inductively coupled plasma detection of the extracted organic silicones. The detection limit for silicones in the final methyl isobutyl ketone sample/extract is approximately 0.01 ppm, and the method is applied to samples from various Japanese rivers to quantify silicones in several environmental materials. Silicones are reported from river waters (up to approximately 50 ppb), riverine sediments (up to 6 ppm), and as an extractable component of fish tissue (up to approximately 0.9 ppm).

A test was developed using Tetrahymena pyriformis in order to determine the toxicity of various chemicals. Pre-cultured T. pyriformis was exposed for 24 h at 30 degrees C to various concentrations of chemicals, and the number of T. pyriformis surviving were then counted. The concentration of the chemical, at which the proliferation of T. pyriformis was restricted to one-half of the blank test (EC50), was determined. The method, applied to 57 chemicals, demonstrated that it could be used to detect the chemicals at low concentrations rapidly and with ease. The EC50 values showed a good relationship with 48 h LC50 values for Himedaka (Orizias latipes), and could be explained on the basis of the partition coefficient between water and n-octanol.

It was found that the water extract of grass-wrack pondweed (Potamogeton oxyphylus Miquel) had inhibitory activity for reverse mutations induced by benzo[a]pyrene, 2-aminoanthracene and 2-nitrofluorene with Salmonella typhimurium TA100 and TA98. The active substance was heat-resistant, and the molecular weight was above 300 000. The substance acted as a desmutagen rather than an antimutagen or inhibitor of metabolic activation.

Although the rates of removal of BOD and other indicators have been studied for the evaluation of night soil treatment plant, there is no report about organic compounds. In order to identify trace amounts of organic compounds in night soil treatment plant at each process stage, the organic compounds in the night soil and in the treated water were subjected to anaerobic methods and analyzed by gas chromatography--mass spectrometry. Cholesterol and coprostanol as fecal sterols, indoles as odor substances and dichlorobenzene, cresol and phenol as disinfectants used in toilets were identified in the sample of night soil. As a result of anaerobic treatment the fecal sterols were degraded, oxindole remained, and phenylacetic acid was identified as a metabolite from tryptophan. These organic compounds were completely degraded by the aerobic treatment when phthalates and adipate were identified in the effluent.

Organosiloxanes (silicones) are being released to the environment in increasing quantities, and there are questions concerning the reactivity of silicones as methylating agents under environmental conditions. Specifically, the interaction of inorganic mercury and polydimethylsiloxanes, and other silicones, to give methylmercury is important in this content. To this end a series of laboratory experiments, in which inorganic mercury, short and long chain silicones, cyclic siloxanes, oxidized silicone gums, and industrial formulations containing silicones, were conducted. In all cases, there was evidence of the production of methylmercury, a finding of potential significance. Analyses for methylmercury and related silicon-containing degradation products were conducted using a combination of gas chromatography, mass spectrometry, and head-space gas analysis.

Phenotypes of alpha-L-fucosidase (Fu) were demonstrated from tissue samples of spleen, liver, lung and kidney stored for a few weeks at room temperature. Activity was fairly low in pancreas, heart, muscle, skin and brain, and typing was not reliable after 1 week storage. Fu types were detectable from dental pulp tissue of up to 1 week storage. Activity was present in hair root cells, but was extremely low. The results show that the Fu typing may be applicable to individual identification of organ tissues and teeth.

1. The protective effect against Cd toxicity of prior exposure to Cd or Zn solutions at low concentration was studied. 2. Carp were bred in tap water (A), 1 ppm Cd solution (B) and 5 ppm Zn solution (C) for 14 days and then transferred into 15 ppm Cd solution. The survival ratio of carp decreased in the order: (C):(B):(A). 3. Binding capacity of Cd to high molecular and metallothionein fractions in the cytoplasmic solutions of the hepato-pancreas was studied and the binding capacity to the metallothionein fraction was stronger than that to the high molecular fraction. The authors recognized that Zn in the metallothionein fraction is substituted by Cd.

The toxicity of xanthene dyes were studied by various interaction between the dyes and the components in vital body. (1) An increase in the amount of Rose Bengale adsorbed on the gill of fish was followed by the increase in red corpuscle number, and it was assumed that the death of fish in xanthene dye solution was due to anoxemia. (2) Binding capacity of xanthene dyes with bovine serum albumin decreased in the following; Rose Bengale, Phloxine, Erythrosine, Eosine and Uranine. This order was quite coincident with the toxicity compared by TLm values. (3) From the results of rec-assay test by use of Bacillus subtilis, it was confirmed that Phloxine and Rose Bengale had DNA-damaging capacity related to the mutagenecity.

Biochemical degradation test of food coal-tar dyes using sludge was studied in order to evaluate their safety. It was found that some dyes were little decomposed under aerobic condition but four azo dyes were readily decomposed under anaerobic condition. These were Food Yellow No. 4, No. 5, and Food Red No. 2, No. 102. Decomposed products of these four azo dyes by sludge under anaerobic condition were identified as sulfonilic, naphthionic acids and so on.

The mechanism of an apparent desmutagenic effect of humic acid was investigated. Firstly, components of humic acid (resorcinol, vanillin, vanillic acid, ferulic acid, protochatechuic acid and benzoic acid) were tested and were not found to show a desmutagenic effect. By contrast, lignin did show a desmutagenic effect. The desmutagenic effect of humic acid was decreased by ozone treatment, and the degree of decrease corresponded with a decrease in KMnO4 consumption. Benzo[a]pyrene and humic acid were incubated at 37 degrees C for 1 h and extracted by ethyl acetate and the extract was investigated by gas chromatography (GC). The peak of the decomposition product did not appear, but the amount of benzo[a]pyrene was decreased. This suggests that the desmutagenic effect of humic acid was caused by adsorption of benzo[a]pyrene by humic acid rather than by decomposition of benzo[a]pyrene. Humic acid had the largest adsorption activity at its critical micelle concentration (CMC), while adsorbed benzo[a]pyrene could be released by ultrasonication. Fulvic acid and water-soluble humic substance showed a slight inhibitory effect on the mutagenicity of benzo[a]pyrene.

Twenty-four strains with high dimethyl disulfide (DMDS)-forming ability were isolated from activated sludge and identified to the genus level. These bacteria were classified into four groups (A, B, C, and D) by the API ZYM System (API System S.A., Montalieu, France). Group A (three strains) was identified as genus Lactobacillus by the API 20B System, by the method of Cowan and Steel, and by production of lactic acid as confirmed by gas-liquid chromatography. Group B (eight strains) was identified as genus Corynebacterium by API 20B and the Cowan and Steel method. Group C (one strain) was suggested to belong to genus Corynebacterium by the API 20B System. Group D (12 strains) was identified as genus Pseudomonas or Alcaligenes by the API 20B System, as genus Alcaligenes by the Cowan and Steel method, and as Achromobacter group Vd by the API 20NE System. However, on the basis of guanine-plus-cytosine contents in DNA and form of flagella, these strains were identified as genus Pseudomonas. Formation of DMDS from DL-methionine and S-methyl-L-cysteine was tested. DMDS-forming bacteria isolated from activated sludge formed DMDS from both precursors. In genus Pseudomonas, P. aeruginosa could not form DMDS from either precursor, but P. acidovorans, P. alcaligenes, P. pseudoalcaligenes, and P. testosteroni formed DMDS. In genus Alcaligenes, A. denitrificans subsp. xylosoxydans, A. denitrificans subsp. denitrificans, A. faecalis, and A. odorans formed DMDS from both precursors. Achromobacter group Vd formed DMDS from S-methyl-L-cysteine, but could not from DL-methionine.

The acute toxicity of photodecomposed products of Erythrosine, Eosine, Phloxine and Rose Bengale were studied, since it was found that toxicity of these dyes to fish increased after the dyes had been photoirradiated. Photodecomposed products of the dyes were isolated and identified with UV, IR, NMR spectra and the acute toxicity of those compounds were determined by TLm test. As results of these studies, it became clear that the toxicity of photodecomposed organic products (dehalogenated compounds of dyes) were lower than the mother compounds. The increases in toxicity of the xanthene dyes by photo-irradiation were attributed to the liberated halogens by irradiation.

The sediments of Nagara river and its bystreams were extracted with ether. The mutagenicities of the extracts were determined by the Ames test with Salmonella typhimurium TA100 with S9 mix. The extracts were fractionated on an activated silica-gel column with four kinds of organic solvent, namely iso-octane, iso-octane:benzene (1:1), benzene:ethyl acetate (1:1) and benzene:methanol (1:1). The highest mutagenicity was observed in the iso-octane-benzene fraction of Arata river by using TA100 with S9 mix. Chemical substances in this fraction were identified by GC-MS and GC, and several kinds of polycyclic aromatic hydrocarbons (PAH) were found. Several peaks were unidentified, but these PAH, especially benzo [b] fluoranthene and benzo [a] pyrene, may contribute substantially to the mutagenicity of this fraction.

The genetic polymorphism of C6 was investigated in 329 unrelated Japanese individuals using isoelectric focusing in polyacrylamide gels followed by an electroimmunoblotting technique. Besides six common phenotypes C6 A, AB, B, AB2, BB2 and B2, six rare variants were observed. The allele frequencies were: C6*A = 0.4422, C6*B = 0.4757, C6*B2 = 0.0714, C6*A3 = 0.0015, C6*M1 = 0.0046 and C6*B3 = 0.0046. The population data confirmed that the C6*B2 allele is the third common allele characterizing Japanese. The present electroimmunoblotting technique was applied to demonstrate C6 types in dried bloodstains. The C6 types were determined from bloodstains stored at 4 degrees C for up to 10 weeks, at room temperature for up to 2 weeks and at 37 degrees C for up to 4 days. The results show that this component system offers a new powerful means for the medico-legal grouping of bloodstains.