There are several theories for genesis of aging mechanisms, among them the error catastrophy theory is most acceptable for the progressive accumulation of changes in DNA with time associated with the ever-increasing likelihood of disease and death that accompanies advancing age. Free radical theory of aging may well chemically explain the changes including DNA, endocrine, immune response, and collagen bridges. Free radical reactions, ubiquitous in living systems, are of both enzymatic and non-enzymatic origin, both being related each other. The latter involves the copper and iron catalyzed reactions of oxygen with organic compounds, which from reactive intermediates serving to bind the multiplicity of further free radical reactions proceeding continuously throughout cells and tissues into a network of reactions with variable degrees of interaction between them. Biological scavenge systems and substances for protecting the progressive radical chain reactions exist in living systems. The relation of progress in the chain to protective mechanisms may decide aging phenomenon. If the biological scavenge system is progammed throughout life, nutrients for scavenging the chain, including vitamin E, may be available for maintaining healthy aging.

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Recent progress of forensic toxicology of morphine and heroin, and of the boundary area was reviewed. Topics included in this review were 1) opioid peptides, 2) opioid receptors, 3) endogenous morphine, 4) metabolism of morphine : glucuronidation, sulfate conjugation, N-demethylation and dehydrogenation, 5) pharmacological activity of morphine conjugates, 6) metabolism of heroin, 7) analytical methods of morphine and heroin for forensic toxicological purpose.

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Disinfectants frequently used in hospitals are presumed to be degraded by bacteria after discharge into the environment. In the present investigation, the degradabilities of several disinfectants such as glutaraldehyde (GA), benzalkonium chloride (BC) and chlorhexidine digluconate (CG) were examined by using three strains of Pseudomonas aeruginosa isolated from activated sludge and acclimatized to the disinfectants. It was found that GA was metabolized to glutaric acid, BC was metabolized to decabutyldimethylamine and toluene, and CG was converted to p-chloroaniline, p-chlorophenol, p-chloroacetoanilide, phenol, aniline, pyrocatechol and pyrogallol. Possible degradation pathways of CG are presented.

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Seven peaks showing glutathione S-transferase (GST) activity toward 1-chloro-2, 4-dinitrobenzene (CDNB) were separated from rabbit liver preparations by CM-52 cellulose column chromatography (pH 6.7). These peaks were designated as R1, R2, Rx, Ry, R3, Rz, and R4 in order of elution from the column. Among these peaks, R3 was found to have notable GST activity, which was responsible for the enzymatic formation of S-methylglutathione from fenitrothion [O, O-dimethyl O-(3-methyl-4-nitrophenyl)phosphorothioate] in the presence of glutathione. The GST (s) in peak R3 gave a characteristic zymogram after agar-gel electrophoresis as compared with other CM-52 column fractions.

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As a result of the study on the environmental pollution by benzo[α]pyrene (BP) from asphalt, the following results were obtained. 1) Acidic alumina (1% water) column treatment was effective to remove co-existing interferences in the determination of BP in asphalt, and enabled to determine ng order of BP by high-performance liquid chromatography-fluorescence detection procedure. 2) BP in the environmental air on asphalting road (about 10 cm above and asphalt temp. was about 100°C) ranged 2.12-17.88 ng/m³ (control 0.30-1.50 ng/m³), while BP in asphalt ranged 1.68-5.20μg/g. No correlation was observed between BP in the air and that in asphalt. 3) BP volatilized from asphalt (BP content 10.87μg/g, surface area 95 cm²) depended on asphalt temperature. The amount volatilized in creased exponentially at the temperature more than 200°C.

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Quantitative screening method for bicozamycin in chicken egg and meat by fluorescence densitometry with high performance thin layer chromatography (HPTLC) was developed. Bicozamycin was extracted with 2% phosphotungstic acid from chicken egg and meat. The extract was cleaned up by a XAD-4 resin column with 60% methanol as an eluent. The eluate was evaporated to dryness, and the residue was dissolved in methanol and acetone. The solution was spotted on a silica gel HPTLC plate. After development, the plate was dipped in 2% sulfuric acid-ether solution and heated. The spots of bicozamycin gave blue fluorescence under long wavelength ultraviolet (UV) light. The fluorescence intensity was determined by densitometry. The calibration curve was rectilinear in the range from 3 ng to 30 ng. The average recoveries of bicozamycin added to chicken egg and meat were 91.8% and 91.4%, respectively. The present method will be applicable to determine bicozamycin with the detection limit of 0.2μg/g.

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Normal phase and reversed-phase thin-layer chromatographic (NP-and RP-TLC) methods were developed for the simultaneous determination of 45 organophosphorus pesticides (OP ; 8 phosphates, 14 phosphorothioates, 15 phosphorodithioates and 8 miscellaneous OP), which were used or had been used in Japan. The pesticides were analysed by three TLC systems by using Merck Silica gel 60 F-254 plate (n-hexane-acetone 7 : 3, v/v) and Merck RP-8F₂₅₄ s and Whatman KC18F plates (methanol-water 7 : 3, v/v). The locations of the pesticides were detected by UV absorption and by colorations with palladium chloride and 4-(p-nitrobenzyl)pyridine (NBP). All of the pesticides tested could be detected with the NBP reagent, and its detection limit was less than 0.25-0.5μg of OP except 1μg of trichlorfon. The most effective discriminating powers (DP) for TLC analyses of OP were given by the combination of two TLC systems with silica gel and chemical bonded silica gels (DP=0.93 and 0.94, numbers of samples 45, and error factor 0.05). Together with our previous papers on analyses of OP by gas-liquid chromatography and directinlet and gas-chromatography-mass spectrometry (DI-and GC-MS), the most reliable procedure proposed for the examination of OP is as follows. At first, samples are applied to the TLC analyses for the screening test of OP, and then the pesticides are identified with GC-MS on column of OV-1 or QF-1 and/or DI-MS.

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The concentration of metallothionein in the mouse tissues was determined after starvation, and the effect of starvation against acute cadmium toxicity was examined. The concentration of metallothionein increased significantly in the liver (101±30μg/g) and pancreas (595±94μg/g) 24 h after starvation. The starved mice were injected s.c. with cadmium at the dose of 10, 8 or 7 mg/g, and observed for 3 or 2 d. The mortality of starved mice were not lower than that of control. The increase of the metallothionein concentration in the liver after sham-operation was also not effective against acute cadmium toxicity. At 8 h after the cadmium injection, the concentration of metallothionein in the liver of starved mice was significantly lower than that of mice pre-injected with olive oil. These results indicate that acute cadmium toxicity in starved or wounded mice can not be prevented even if the concentration of metallothionein in the liver increased.

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For simple and quick determination of paraquat [PQ] in biological material, availability of supernatant obtained by protein precipitation directly for the colorimetric determination after reduction of PQ with alkaline sodium hydrosulfite was examined. Interference of four protein precipitants commonly used were tested for alkaline sodium hydrosulfite reduction of PQ. The characteristic spectrum of reduced form of PQ changed in the presence of trichloracetic acid or perchloric acid. Tungstic acid, under acidic condition, reacted with alkaline sodium hydrosulfite and developed dark blue colour. When sample was first adjusted to alkaline pH by NaOH solution and then crystaline sodium hydrosulfite was added, formation of tungstic blue was, however, not seen. Use of these three protein precipitants might, therefore, lead to misunderstanding in PQ determination and should be avoided. In contrast, sulfosalicylic acid showed no interference in the following determination of PQ and PQ added to homogenate was recovered quantitatibly. Chosing a suitable precipitant, sulfosalicylic acid, post protein precipitate supernatant was available for the determination of PQ directly in tissue homogenate.

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Regression analysis was carried out in order to predict the effective concentration (EC₅₀) of 34 chemicals for inhibition of the growth of Tetrahymena pyriformis in 24 h. We examined 4 items (n-octanol/water partition coefficient (P), molecular weight (M_r), organic and inorganic characters, and molecular connectivity indices) and their derivatives. The best one-parameter equation was that involing M_r [log EC₅₀=-1.964 (M_r/100)+5.380 (r=-0.821, s=0.603, n=34)], and the second best involved the 3rd order valence molecular connectivity index of path type, ³χ^v_p[log EC₅₀=1.108 ³χ^v_p+3.944(r=-0.810, s=0.619, n=34)], while the n-octanol/water partition coefficient was least effective for the prediction of EC₅₀ values [log EC₅₀=-0.735 log P+4.233 (r=-0.780, s=0.660, n=34)]. The best combination for a two-parameter equation was ³χ^v_p and M_r [log EC₅₀=-1.105 (M_r/100)-0.638 ³χ^v_p+4.990 (R=0.866, s=0.516, n=34)]. Regression analysis with the same 4 items on median lethal concentration (LC₅₀) of 123 chemicals for Oryzias latipes (red killifish) was refered and the limitations in the application of the parameters and the equations botained are discussed.

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To study benzo[α]pyrene (BP) pollution on traffic-related environment, the contribution of gasoline engine car was investigated. BP was extracted by ultrasonication (only exhaust gas), liquid-liquid partitions, followed by a clean-up with Sep-Pak C₁₈, and determined by reversed phase high-performance liquid chromatography with a fluorescence detector. The BP content in lubricating oils increased with an increase in used periods of these oils (max. 5.32μg/ml), though no correlation was observed between the contents of BP in lubricating oils and those in exhaust gas. BP in exhaust gas was very low except one that has no catalytic converter. It was found that the catalytic converter decreased or degraded more than 90% of BP in exhaust gas. It is also suggested that the effect of BP adsorbed on soot on the contents of BP in exhaust gas might increase with an increase in used periods of cars. The experiment with new engine demonstrated that BP contents in exhaust gas had little correlation with those in lubricating oils or gasoline.

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A simplified method with Sep-Pak silica cartridge (Waters Associates) for the determination of polycyclic aromatic hydrocarbons (PAHs) (i.e. anthracene (An), pyrene (Py), benzo[a]anthracene (B[a]A), benzo[b]fluoranthene (B[b]F), benzo[k]fluoranthene (B[k]F), benzo[a]pyrene (B[a]P), benzo[g, h, i]perylene (B[g, h, i]P) in daily diet has been developed. The method consists of alkali digestion with sodium sulfide as an antioxidant, extraction with n-hexane, clean-up with Sep-Pak silica cartridge, determination with high-performance liquid chromatography. Recoveries (%) and coefficientsof variation (C.V.%) of 7 PAHs (50 ng-200 ng) added to 50 g of homogenized daily dist were 90.2% (An)-102.7% (Py) and 2.97% (B[a]A)-7.56% (B[a]P), respectively.

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Airborne particles, airborne microorganisms, microorganisms fallen down on agar plates and those on the surfaces were determined in the surgical operation rooms and preceding rooms of public and private hospitals in Tokai area. Microorganisms detected in these places were classified morphologically. In general, the number of airborne particles (≥0.5μm) determined by a light scattering particle counter was in a relatively good correlation with that of airborne fungi determined by the impinger method, which was closely related to the conditions of air-conditioning of these rooms. On the contrary, no significant correlations were observed between the number of airborne particles and the number of airborne bacteria determined by three different methods.

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