Green tea (GT) drinking showed chemopreventive effects on various cancers. In addition, inhibition of CYP1A activity by green tea components—polyphenols—has been suggested as a chemoprevention against carcinogens that were bioactivated by CYP1As. Therefore, any changes in hepatic CYP1As may be considered as a biomarker for GT chemoprevention and clarify whether whole GT is chemopreventive for the population who are exposed to CYP1A specifically-bioactivated carcinogens. In this study, we investigated the changes in CYP1A levels by pre- and concurrent GT drinking against a CYP1A-inducing carcinogen, 3-methylcholanthrene (MC), in aryl hydrocarbon receptor responsive C57 BL/6 mice. We found that GT drinking itself induced hepatic CYP1As and enhanced MC-induced ethoxyresorufin-O-demethylase (EROD) activity (P<0.05). However, our studies of CYP1A monoclonal antibody and western blots revealed that the enhanced hepatic EROD activity by GT did not come from CYP1As. Therefore, our results suggest that GT may work to biotransform CYP1A inducing carcinogens into non-carcinogenic metabolites by modulation of other microsomal enzymes rather than CYP1As. In addition, the mechanism of GT chemoprevention may be different from that of GT components, such as polyphenols that reduce CYP1As activity.

There are few reports regarding the effects of benzene and ethanol being administered simultaneously. In our experiments with 4 groups (controls, ethanol, benzene and ethanol plus benzene) Wistar male rats were treated with ethanol (20%) for 5 weeks, and then exposed to benzene (0.26 g/kg) for 5 days per week for 3 weeks. We also investigated the effects of benzene on the body weight, organ weight, peripheral hematology and hepatic drug metabolizing enzymes in the ethanol administrated rats. The liver weight increased significantly, but spleen weight decreased significantly in the benzene exposed group. Hematological examination showed a decrease of leukocyte in the two groups of benzene and ethanol plus benzene in comparison with the controls, but an effect promoted by ethanol was not found. Serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) values were not significantly different in the exposure groups when compared with the controls. The contents of microsomal cytochrome P450 in all the exposed groups showed a tendency to increase, but they were not significantly different in comparison with the controls. On the other hand, hepatic glutathione S-transferase activity in the exposed groups increased significantly.

We investigated the air pollution in the student smoking hall from February 1st in the winter season, and during 8 days in the middle of April in the spring season. The student smoking hall was in an open, draught location. Moreover, the concentration of pollutants were measured in three time periods: break, lecture and lunch times. The pollutants measured were nitrogen monoxide (NO), nitrogen dioxide (NO2), carbon monoxide (CO), carbon dioxide (CO2), suspended particulate matter (SPM) and polynuclear aromatic hydrocarbon (PAH). The concentrations of pollutants measured during the break times were of a relatively higher level than those during the lecture and lunch times. However, the concentration of pollutants were not influenced by ventilation operation in the smoking hall, which is not a closed place. SPM and CO2 concentrations during the break time were instantaneously investigated above Building Sanitation Standards Management, Japan (SPM: 0.15 mg/m3, CO2: 1,000 ppm). Especially, the SPM concentration was recognized to be strongly influenced by tobacco smoke.

t(11;14)(q13;q32) observed in B‐cell malignancies is associated with cyclin D1 (bcl‐1, PRAD1, CCND1) overexpression. We devised a simple competitive reverse transcription‐polymerase chain reaction (RT‐PCR) assay for rapid detection of cyclin D1 overexpression. Sharing a single upstream primer derived from a homologous sequence in cyclins D1, D2 and D3, each PCR product serves as a competitor and cyclin D1 overexpression is determined by comparing the intensities of the three amplified products. We analyzed cyclin D1 in clinical specimens from 104 patients with lymphoid malignancies. Cyclin D1 overexpression was evident in 13 of 104 (7/72 non‐Hodgkin's lymphomas, 0/6 adult T‐cell lymphoma/leukemias, 0/4 Hodgkin's diseases, 0/11 acute lymphoblastic leukemias, 3/4 multiple myelomas, 1/2 Waldenströmas, macroglobulinemias, 1/2 prolymphocytic leukemias and 1/3 chronic lymphocytic leukemias). Among 72 patients for whom cytogenetic studies had been done, all 7 patients with t(11;14) were positive. The relative expression levels of D‐type cyclins altered dramatically in the presence of t(11;14). Thus, this RT‐PCR assay can identify tumors with cyclin D1 overexpression. Cyclin D1 overexpression was frequent in extranodal specimens (11 out of 32 vs. 2 of 72 lymph nodes) and was restricted to specific types of lymphoid malignancies, as observed using other methods. This reliable assay should be suitable to provide clinical guidance for the diagnosis and management of lymphoid malignancies, especially in the case of extranodal involvement.

The retinoblastoma tumor suppressor (Rb) gene product plays an essential role in cell-cycle regulation. However, its role in terminal differentiation of hematopoietic cells is speculative. Here we show a model of 12-o-tetradecanoylphorbol-13-acetate (TPA)-induced hematopoietic differentiation and growth arrest with a defective Rb-mediated pathway. TPA treatment arrested the cell cycle of a human hematopoietic cell line, MEG-01s, at the G1-S boundary and induced expression of p21/SDI1/WAF1/CIP1 and p27/KIP1. Both of these proteins were present in cyclin E-associated complexes, the histone H1 and Rb kinase activities of which were then inactivated. However, MEG-01s cells lacked the intact Rb protein and the Rb-mediated pathway was defective. This model raises a question about the role for Rb in terminal differentiation of hematopoietic cells.

A rare outbreak of acute hepatic damage in workers exposed to dichloropropanols (DCPs) was reported recently. The purpose of the present study is to examine the effects of DCPs on various organs, the dose dependency and the pathogenetic potential of DCP hepatotoxicity. A single intraperitoneal injection was given to six groups of rats with 0.2 ml of 20% ethanol (control), or 1/8 x, 1/4 x, 1/2 x, 1 x, and 2 x LD50 (0.11 ml/kg) of 1,3-dichloro, 2-propanol (DC2P) diluted in 20% ethanol. After blood samplings, all organs were subjected to histologic examinations with light and electron microscopes. Fresh liver tissues from further 4 control and 4 experimental rats sacrificed 6 hours after the injection of 20% ethanol and 1 x LD50 of DC2P were homogenized and subjected to evaluate lipid peroxidation, glutathione S-transferase activity and reduced glutathione contents in the liver. The rats administered with only ethanol or 1/8 and 1/4 LD50 of DC2P showed no serological or histopathological abnormalities. Marked elevation of serum glutamate pyruvate transaminase (SGPT) with a diffuse massive necrosis of the liver cells were noted in all rats of both the 1 x and 2 x LD50 groups, and irregular zonal necroses were found in 3 of 4 rats injected with 1/2 LD50. There were no serious toxic changes in other organs. Hepatic malondialdehyde level was significantly increased, associated with decreases of liver glutathione S-transferase activity and reduced glutathione content. It was concluded that the serious DC2P-toxicity was mainly found in the livers, dose dependently, and one of the causative mechanisms of this hepatotoxicity might be the lipid peroxidation.

After the intravenous administration of 14C-D-aspartic acid (Asp) into Sprague-Dawley rats (male, 7-week-old), the distribution and elimination of radioactivity was investigated by the whole body autoradiography. High radioactivities were detected in pineal gland, pituitary gland and salivary gland at 30min after administration. The other tissues detected were liver, lung, adrenal gland, pancreas and spleen where D- Asp was reported to occur naturally. After 24hr, the radioactivities were still detected at high levels in the pineal, pituitary and salivary glands. The data suggested the natural occurrence of D-Asp in salivary gland. After careful examination utilizing fluorescent derivatization and chiral separation by high-performance liquid chromatography, the presence of D-Asp was, for the first time, demonstrated in salivary gland in situ, the concentration of which was 7.85±1.0nmol/g. The administration of 14C-L-Asp was also carried out. The data suggested that D-Asp in the circulating blood is one of the sources of the tissue D-Asp.

Specific polyclonal antibody was raised against D-aspartate (D-Asp) which had been conjugated to glutaraldehyde and was purified by affinity chromatography. Immunohistochemical staining of rat pineal gland with the antibody demonstrated the presence of D-Asp in the cytoplasm of pinealocytes, the predominant cell type in this gland. D-Asp immunoreactivity was more evident in the distal region than in the proximal region of the gland. Pinealocytes in the distal region are presumably involved in the synthesis and secretion of the pineal hormone, melatonin, and the results of staining may indicate some yet unknown role of D-Asp in the regulation of melatonin secretion.

The concentrations of indoor pollutants generated from types of heaters were measured in a model room of 20m2 in area and 45m3 in capacity. We used six different heaters: three kerosene heaters of different types, town and propane gas heaters, and an electric heater. Three ventilation conditions were introduced into each experiment: non-ventilation, fan-on ventilation with closed door and fan-off ventilation with half-opened door. The results obtained by heating under non-ventilation condition were as follows: The concentrations of NO2 and CO2 were comparatively high and the values obtained from all the heaters except the electric heater exceeded the 1-hr Environmental Quality Standards, Japan (EQS NO2: 0.04-0.06 ppm) and the Building Sanitation Management Standards, Japan (BSMS CO2: 1,000 ppm), respectively. The CO concentration emitted from reflection kerosene and town gas heaters slightly exceeded the BSMS (10 ppm). The concentrations of suspended particulate matter and polynuclear aromatic hydrocarbons showed an increasing tendency during the use of kerosene-fueled heaters. Under two ventilating conditions, NOx concentration decreased to less than a third in comparison with non-ventilating condition.

A simple analytical method was developed to determine p-nitrochlorobenzene (p-NCB) in working environmental air by reversed-phase high-performance liquid chromatography (HPLC). Nitrobenzene, nitrotoluene isomers, and NCB isomers were separated and p-NCB was determined by reversed-phase HPLC as follows: column; Hitachi gel #3056 (4.0 mm I.D. x 150 mm), mobile phase; 2-propanol: water (40:60, v/ v) with a flow rate of 0.8 ml/min, column temperature; 50 degrees C, detector; UV detector at 270 nm. Nitrobenzene, o-, p-nitrotoluene, and o-, m-NCB were separated from p-NCB, but m-nitrotoluene could not be completely separated from p-NCB. The limit of detection (signal-to-noise ratio of 3:1) of p-NCB was 0.3 ng. Repeatability of the HPLC method was excellent. The HPLC method enabled to determine one-tenth of the administrative level (1 mg/m3) of p-NCB, since the lower limit of determination was 0.05 mg/m3. Therefore, the present HPLC method proved to be applicable to the determination of p-NCB in working environmental air.

The relationship between occupational exposure to methyl ethyl ketone (MEK) and its concentration in urine and blood was studied in a group of 72 workers in a printing factory. Personal exposure monitoring was carried out with passive samplers during the workshifts. The time weighted average (TWA) concentration of MEK ranged from 1.3 to 223.7 ppm, with a mean concentration of 47.6 ppm. In addition to MEK, toleuene, xylene, isopropyl alcohol, and ethyl acetate were detected as the main contaminants in all samples.

In order to investigate the relationship between aryl hydrocarbon hydroxylase (AHH) activity and smoking or lung cancer, AHH activities in fresh lungs (normal tissue, tumorous tissue, and surrounding tissue of tumor) obtained from lung cancer patients and non-lung cancer patients were measured. There were no differences in lung AHH activity in the lung lobes. In the non-lung cancer patients, AHH activities ranged from 0.13 to 2.37 (pmol 3 hydroxybenzo[a]pyrene/20 min/mg protein), and whereas in the normal tissues of the lung cancer patients they ranged from 0.19 to 5.05. Lung AHH activities showed normal distribution, and a large variation (26 times) was observed in normal tissues in the lung cancer patients. In most cases, AHH activities in the tumorous tissues and the surrounding tissue of the tumor were lower than those in the normal tissues of the lung cancer patients. In the non-lung cancer group, the means of AHH activity of the nonsmoker subgroup (NN) and the smoker subgroup (SN) were 0.62 and 0.96, respectively. On the other hand, in the lung cancer group the means of AHH activity of the nonsmoker subgroup (NC) and smoker subgroup (SC) were 0.85 and 1.05, respectively. Statistically significant differences were observed between NN and SN, NN and NC, and NN and SC. These results suggest that human lung AHH activity was increased by cigarette smoke as in rodent lungs, and the distribution of basal AHH activity in lung tissue of the nonsmokers group in the lung cancer patients shifted toward high levels compared to the nonsmokers group in the non-lung cancer group. The effect of the histological cell types of the lung cancer on the AHH activity was not observed in this study.

アセトンは産業現場で広く使用されている有機溶剤である.生体内に吸収されたアセトンの一部は,代謝を受けずにそのまま尿中に排泄されることが知られている.アセトンの生物学的暴露指標として,尿中アセトン濃度の測定が広く行われている.尿中アセトンの測定法はこれまで吸光光度分析法,ガスクロマトグラフ分析法,ヘッドスペースガスクロマトグラス分析法が報告されているが,これらの分析法には特異性に欠ける点,種々の技術的問題点や高価な機器を必要とするなど,必ずしもルーチン分析に適しているとは言い難い.そこで著者らは2,4-ジニトロフェニルヒドラジン(DNPH)を用いたアセトンの誇導体化,有機溶媒抽出,逆相高速液体クロマトグラフィー(HPLC)から成る尿中アセトンの分析法を検討した.標準液または尿0.5 mlを共栓試験管にとり, 0.25% DNPH溶液0.4 mlを加えて5秒間撹拌した. 5分経過後, n-ヘキサン3.0 mlを加えてアセトン-DNPH誇導体を抽出した. n-ヘキサン層2.0 mlをとり,濃縮乾固し,アセトニトリルに溶解しHPLCに注入した. HPLCの条件は次のとおりである.分析カラムにはTSK gel ODS80TM (4.6 mm i. d.×150 mm),移動相にはアセトニトリル-水(80:20, v/v)を用い,流速は1.0 ml/minとした. DNPH誘導体の検出には365 nmを用いた.本測定法は尿中に存在し, DNPHと反応するヒドロキシアセトンやアルデヒド及び他のケトン類(メチルエチルケトン,メチルイソブチルケトン)とも完全に分離できることが分かった.検量線は少なくとも2.70 mmol/lまでは直線性を示し,検出下限は1 μmol/lであった.アセトン-DNPH誘導体の安定性を検討した結果, 4℃保存では少なくとも2日間は誘導体の濃度に変化は認められなかった.また,回収率,再現性とも良好な結果が得られた.したがって,プレラベル法を用いた本HPLC法は簡易,迅速で多量の検体を処理するルーチン分析に有用であると思われる.

The effect of one-shot or repeated treatment with methyl tertiary-butyl ether (MTBE) on lipid peroxidation and cytochrome P-450 in mouse liver was examined. One-shot treatment at the 500 mg/kg dose level caused a transient decrease in leukocyte number after 24 h. An increased level of lipid peroxide was observed in the liver homogenates 24 h after one-shot treatment and four weeks of repeated 200 mg/kg treatment. The hepatic cytochrome P-450 content increased after one and four weeks of repeated treatment (50, 200 mg/kg). On the other hand, the treatment did not affect glutathione content and glutathione-S-transferase activity. These results indicate that MTBE treatment caused lipid peroxidation in the liver homogenate and induction of hepatic microsomal cytochrome P-450 content.

A simple method for measuring cleaning solvent in air by gas detector tube for ethyl acetate was evaluated with regard to basic performance such as sensitivity, precision, discolor tone and effect of temperature. Satisfactory results were obtained with regard to sensitivity, precision, discolor tone in the indicator and stability of discolored length. Range of measurable concentration by the present method was about 25-800 mg/m3. It was found necessary in the present method to make temperature correction. It was concluded from these results that the present method is useful for measuring cleaning solvent in air. However, positive effect on discolored length was found by vapor of toluene and ethyl acetate when they coexisted. No effect on discolored length was found by vapor of acetone, methanol or trichloroethylene.

In this paper, we determined whether ethylene glycol monomethyl ether (EGME) and diethylene glycol monomethyl ether (diEGME) induce hepatic gamma-glutamyl transpeptidase activity. Male adult Wistar rats weighing 220 g were used as experimental animals. EGME (100, 300 mg/kg per day) and diEGME (500, 1000, 2000 mg/kg per day) were administered by gavage for 1, 2 or 5 days or 4 weeks. In the 4-week study, experimental animals were administered EGME or diEGME once a day orally, 5 days/week. EGME treatment increased the serum gamma-glutamyl transpeptidase (GGT) level significantly, however, diEGME did not. The activities of three other enzymes (SGOT, SGPT and ALP) in serum were not altered by EGME or diEGME treatment and thus there was no biochemical indices of hepatic damage by EGME or diEGME. EGME treatment increased the GGT activities in the liver and lungs. Of the organs examined, the induction of GGT was the greatest in the liver. The inducibility in the liver was 216% for the 5-day treatment and 460% for the 4-week treatment. A dose-dependent increase of hepatic microsomal GGT activity by EGME was observed. On the other hand, renal GGT activities were declined to 72% and 60% of control by the 5-day and 4-week EGME treatments, respectively. DiEGME did not affect the GGT activities in any of the tissues except those of the brain. In the histochemical study, most hepatocytes at the periportal zones were stained with GGT staining after the 4-week treatment. However, the hepatocytes at the central zones were negative.