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In the present study, effectiveness of topical vidarabine or subsequent 5-fluorouracil (5-FU) administration was examined against persistent genital human papillomavirus (HPV) infection after local surgery. Thirty patients underwent local eradication treatment of uterine cervical intra-epithelial neoplasia (CIN) and stage Ia1 uterine cervical cancers. HPV typing was performed by PCR-RFLP analysis. HPV infection was detected pre-operatively in 29 of 30 patients. Of these, HPV was still present in the 20 patients within two months after the therapy. Topical administration of vidarabine or subsequent 5-FU once a week for four weeks was performed to the post-operative persistent HPV-positive cases. HPV infection was abolished in 1 of 10 (10%) with topical vidarabine, and in 2 of 4 vidarabine-resistant cases (50%) with topical 5-FU. Topical vidarabine or 5-FU treatment is beneficial for HPV-positive cases after local surgical excision.

A cultivation procedure for the preparation of a new tempeh-like fermented soybean containing a high level of gamma-aminobutyric acid was developed. Steamed soybeans were incubated aerobically with Rhizopus microsporus var. oligosporus IFO 8631 for 20 h, and then anaerobically incubated for 5 h by replacement of the atmosphere with nitrogen. The GABA content in the aerobically fermented soybeans was about 30 mg per 100 g dry fermented soybeans, while the anaerobically cultivation was about 370 mg/100 g dry fermented soybeans. The incubation with several strains of Rhizopus species showed that all of R. microsporus var. oligosporus and R. oryzae examined accumulated GABA in the anaerobically fermented soybeans. In particular, R. microsporus var. oligosporus IFO 32002 and IFO 32003 showed the highest content of GABA (1,740 mg/100 g dry fermented soybeans and 1,500 mg/100 g dry fermented soybeans, respectively). Moreover, the free protein amino acids increased greatly in the fermented soybeans during the anaerobic cultivation.

We previously reported the special herbal complex (Hoelen, Angelicae radix, Scutellariae radix and Glycyrrhizae radix) suppressed telomerase activity in chemo-endocrine-resistant cancer cell lines. The present study attempted to determine whether the above herbal complex induces apoptosis in endocrine-resistant AN3CA and adriamycin-resistant MCF7/ADR carcinoma cells. Exposure to the herbal complex decreased cell viability in a time- and dose-dependent manner in the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The agent induced cellular apoptosis was determined by DNA fragmentation and a nuclear staining assay. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) showed the decreased expression of apoptosis-related genes, bcl-2, c-myc and human telomerase catalytic subunit (hTERT). A decreased protein level of bcl-2 and c-myc was also determined by Western blot analysis. The data imply that the decreased expression of the genes via suppressing telomerase activity is involved in cellular apoptosis in endocrine-resistant AN3CA cells. Thus, it is suggested that the special herbal complex may be a promising candidate for the treatment of endocrine-resistant gynecologic carcinomas.

The purpose of this study was to investigate negative bias on self-referent processing in depression, focused on the mood congruent effects in a natural depressed state and an experimentally induced transient depressed mood state. In Experiment 1, autobiographical memories and self-relevant ratings of personality trait words were examined in a natural depressed state or non-depressed state, which were measured by Beck Depression Inventory. Results revealed the mood congruent effects on both tasks. In Experiment 2, the same tasks as Experiment 1 were conducted in a transient depressed mood state or non-depressed mood state, which were induced through listening music. Unlike Experiment 1, there were no effects in both tasks, and a positive bias was observed in both mood states. It was suggested that transient mood state did not bias self-referent processing in depression, and Beck's schema hypothesis was supported.

We have found that Juzen-taiho-to has a preventive effect on endometrial carcinogenesis in mice (Carcinogenesis 22 (2001) 587). In the present study, the constituents of Juzen-taiho-to responsible for this effect were explored using a short-term experiment. Thirty female ICR mice were divided into five groups: Group 1 was given a diet containing 0.2% of Juzen-taiho-to and 5ppm estradiol-17beta (E(2)); Group 2 was given a diet containing Shimotsu-to (0.07%) and E(2) (5ppm); Group 3 received Shikunshi-to (0.08%) and E(2) (5ppm) in the diet; Group 4 was given 5ppm E(2) in the diet; and Group 5 served as a control. Exposure of Juzen-taiho-to or Shimotsu-to decreased E(2)-stimulated expression of estrogen-related gene c-fos mRNA (P<0.05), and the cytokines interleukin-1alpha mRNA and tumor necrosis factor alpha mRNA P<0.01). A similar trend was not found upon treatment with Shikunshi-to. These findings suggest that Shimotsu-to is responsible for the inhibitory effects of Juzen-taiho-to on the estrogen-related endometrial carcinogenesis in mice.

Short- and long-term experiments were designed to determine the effects of toremifene (TOR) on estrogen-related endometrial carcinogenesis in mice. In the short-term experiment, a single low dose of TOR (0.2 mg / 30 g body weight) decreased expression of c-fos, interleukin (IL)-1alpha, estrogen receptor (ER)-alpha mRNAs and corresponding proteins induced by estradiol-17beta (E(2)), in the uteri of the ovariectomized mice. Expression of ER-beta mRNA was increased by the TOR treatment, compared with the control. In the long-term experiment, 106 female ICR mice were given N-methyl-N-nitrosourea (MNU) into their uterine corpora. The animals were divided into four groups as follows: group 1, E(2) diet (5 ppm) plus TOR (0.2 mg / 30 g body weight, subcutaneously, every four weeks); group 2, E(2) diet alone; group 3, basal diet plus TOR. Group 4 served as the control. TOR treatment decreased the incidence of MNU and E(2)-induced endometrial adenocarcinoma and atypical hyperplasia at the termination of the experiment (30 weeks after the start). These results suggest that TOR exerts preventive effects against estrogen-related endometrial carcinogenesis in mice, through the suppression of c-fos as well as IL-1alpha expression induced by E(2). Such suppressive effects of TOR may be related to the decreased ER-alpha and increased ER-beta expressions.

To evaluate the impact of the 1999 criticality accident in Tokai-mura on the U isotope composition in soils, U isotopes (235U and 238U) were determined with inductively coupled plasma-mass spectrometry (ICP-MS) for soil core samples collected on the JCO grounds after the accident. The 235U/238U ratios were higher than the natural ratio in most samples. The highest ratio observed was 0.0262. Although vertical profiles of the 235U/238U ratio differed among the soil cores, the ratios tended to be high at the surface and decreased with depth. The U concentration also changed with depth. The percentages of 235U in the excess U, estimated from the positive correlation between U concentration and the 235U/238U ratio in soil samples, were less than 4% by mass (mostly 1-3%) and were much lower than the enrichment of the U used in the uranium conversion building at the time of the criticality accident (18.8%). These findings indicate that enriched U had been released before the criticality accident during the U processing at JCO in connection with the reconversion of light water reactor fuel. Since the range of the U concentrations found was comparable to the range of uncontaminated Japanese surface soils, the amount of U added to the soil was judged negligible from a radiation protection viewpoint.

Nuclear weapons tests conducted by the United States in the Marshall Islands produced significant quantities of regional or tropospheric fallout contamination. Here we report on some preliminary inductively coupled plasma-mass spectrometry (ICP-MS) measurements of plutonium isolated from seven composite soil samples collected from Bikini, Enewetak and Rongelap Atolls in the northern Marshall Islands. These data show that 240Pu/239Pu isotopic signatures in surface soils from the Marshall Island vary significantly and could potentially be used to help quantify the range and extent of fallout deposition (and associated impacts) from specific weapons tests. 137Cs and 60Co were also determined on the same set of soil samples for comparative purposes.

In the Tokai-mura criticality accident, three workers were heavily exposed. Biological materials, such as blood, urine, vomit and hair, were collected from the workers and analyzed for radioactivities, produced by the neutron irradiation. Activation products. such as 24Na, -K and 82Br, were found in these materials by gamma-ray spectrometry. The radionuclide of the highest activity observed in biological materials was 24Na, e.g. the concentrations of this nuclide in the blood samples from the three patients at the accident time were 169, 92 and 23 Bq/ml, respectively. The concentrations of stable sodium in the same samples were determined by ICP-AES to obtain specific activities of 24Na (concentration ratio between the produced 24Na and stable 23Na), which are essential for estimating the neutron fluences and radiation doses. The specific activities of 24Na obtained for the three patients through the blood analysis were 8.2 x 10(4),4.3 x 10(4) and 1.2 x 10(4) Bq24Na/g23Na. Based on these values, individual's neutron fluences were estimated to be 5.7 x 10(11), 3.0 x 10(-1) and 0.85 x 10(11) cm(-2), respectively.

The effects of isoflavones (genistein and daidzein) on endometrial carcinogenesis in mice were investigated in two experiments. In the short-term experiment (2 weeks), single subcutaneous (s.c.) administration of genistein [1 mg / 30 g body weight (b.w.)] significantly decreased the levels of estradiol-17beta (E(2)) (5 ppm in diet)-induced expression of c-jun, interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha) mRNAs in the uteri of ovariectomized mice (P < 0.005, P < 0.05 and P < 0.01, respectively). Daidzein significantly inhibited E(2)-induced expression of c-fos and IL-1alpha (P < 0.01, P < 0.01 respectively). In the long-term experiment (30 weeks), 140 female ICR mice were given N-methyl-N-nitrosourea-containing solution (1 mg / 100 g b.w.) and normal saline (as controls) into their left and right uterine corpora, respectively. They were divided into six groups; group 1 was given E(2) (in diet) alone. Group 2 was given E(2) and genistein (1 mg / 30 g b.w., s.c., every four weeks). Group 3 was exposed to E(2) and daidzein (1 mg / 30 g b.w., s.c., every four weeks). Groups 4 and 5 respectively received genistein and daidzein, and were kept on the basal diet. Group 6 was kept on the basal diet and served as a control. At the termination of the experiment, incidences of endometrial adenocarcinoma and atypical endometrial hyperplasia of the group given E(2) and genistein or daidzein were significantly lower than of the group with E(2) alone (P < 0.01 and P < 0.05, respectively). It is suggested that both genistein and daidzein have an inhibitory effect on estrogen-related endometrial carcinogenesis in mice, possibly by suppressing expression of estrogen-induced estrogen-related genes c-fos and c-jun, and internal cytokines IL-1alpha and TNF-alpha through a cytokine and estrogen receptor-mediated pathway.

In the course of studies on novel retinoids, we have designed and synthesized a series of quinoline derivatives. One of them, 4-[5-[8-(1-methylethyl)-4-phenyl-2-quinolinyl]-1H-2-pyrrolyl]benzoic acid (12f) shows potent RARalpha-selective antagonistic activity.

Two experiments were conducted to determine effects of Juzen-taiho-to on endometrial carcinogenesis in mice. In the first experiment, Juzen-taiho-to treatment (2 weeks) decreased the levels of estradiol-17beta (E(2))-stimulated expression of c-fos/jun mRNA and their oncoproteins, determined by reverse transcription-polymerase chain reaction and Southern blot analysis, and the immunohistochemical method, in the uteri of ovarectomized mice. For the second experiment, 93 female ICR mice were given N-methyl-N-nitrosourea (MNU) solution (1 mg/100 g body weight) and normal saline (as controls) into their left and right uterine corpora, respectively, and were divided into four groups. Group 1 was given a diet containing 0.2% Juzen-taiho-to and 5 p.p.m. E(2). Group 2 was given a diet containing 5 p.p.m. E(2) alone. Group 3 was given a diet containing 0.2% Juzen-taiho-to alone. Group 4 was kept on the basal diet alone and treated as a control. Juzen-taiho-to treatment significantly decreased incidences of the uterine endometrial atypical (P<0.01), complex (P<0.05) and simple hyperplasias (P<0.01), under estrogenic stimulation. It is suggested that Juzen-taiho-to has an inhibitory effect on E2-related endometrial carcinogenesis in mice, relevantly through suppression of estrogen-induced c-fos/jun-expression.

Short and long-term experiments were designed to determine effects of danazol on estrogen-related endometrial carcinogenesis in mice. The short-term assays showed that danazol decreased expression levels of c-fos/jun mRNA and their oncoproteins induced by estradiol-17beta (E2). For the long-term assay, 85 female ICR mice were given N-methyl-N-nitrsourea solution into their uterine corpora. The animals were divided into three groups as follows: Group 1, E2-diet (5 ppm) plus danazol (2 mg/body (s.c.), every 4 weeks); Group 2, E2-diet alone, Group 3, basal diet alone. At 30 weeks, incidences of atypical and complex endometrial hyperplasia were significantly decreased by danazol-treatment. These results suggest that danazol has preventive effects on estrogen-related endometrial carcinogenesis in mice, through the suppression of estrogen-induced c-fos/jun-expression.

In the course of our studies on retinoic acid receptor (RAR) agonists, we have designed and synthesized a series of benzofuran and benzothiophene derivatives. Some of these compounds (1a,b,e,f,j) markedly inhibited LPS-induced B-lymphocyte proliferation and exerted RARalpha selectivity. One of them, 4-[5-(4,7-dimethylbenzofuran-2-yl)pyrrol-2-yl]benzoic acid (1b), when orally administered significantly inhibited mouse antibody production and delayed type hypersensitivity (DTH) responses from a dose of 0.1 mg/kg.

Technetium (Tc) is known to have high mobility in a soil-water system and also high bioavailability for plants, because the most stable form of Tc in natural surface environment is thought to be TcO4- which is highly soluble. The chemical form of Tc, however, changes with environmental conditions. Thus, it is necessary, for realistic assessment, to obtain transfer parameters, such as transfer factors, under natural conditions. However, it is difficult to obtain these parameters using global fallout 99Tc in actual fields due to its low concentration. In this study, 99Tc concentrations in surface soil and plant leaf samples collected from forest sites within the 30-km zone around the Chernobyl reactor were measured for the first time. In the case of soil samples, a simple and rapid analytical method for determination of 99Tc is used which consists of volatilizing and trapping Tc in a combustion apparatus, purifying the Tc with an extraction chromatographic resin, and measuring it by inductively coupled plasma mass spectrometry (ICP-MS). For plant samples, a wet digestion method in combination with the resin is applied and the 99Tc is measured by ICP-MS. Concentrations of 99Tc in organic soil samples and leaves of strawberry (Fragaria vesca) range from 1.1-14.8 Bq kg(-1) dry weight and 0.2-6.0 Bq kg(-1) dry weight, respectively. Our results indicate that soil-to-plant transfer factors for Tc are similar to those for Cs.

Extraction techniques for recovery of technetium-99 (99Tc) for Inductively Coupled Plasma Mass Spectrometry (ICP-MS) measurements were evaluated using soil samples collected from the Marshall Islands. The results of three different extraction techniques were compared: (MI) acid leaching of Tc from ashed soil; (M2) acid leaching of Tc from raw dry soil; and (M3) Tc volatilization from ashed soil using a combustion apparatus. Total Tc recoveries varied considerably between the extraction techniques but each method yielded similar analytical results for 99Tc. Applications of these extraction techniques to a series of environmental samples and ICP-MS measurements have yielded first data on the 99Tc content of Marshall Islands soil samples contaminated with close-in radioactive fallout from nuclear weapons testing. The 99Tc activity concentration in the soil samples ranged between 0.1 and 1.1 mBq g(-1) dry weight (dw). The limit of detection for 99Tc by ICP-MS was 0.17 mBq per sample or 0.014 mBq g(-1) dw under standard operating conditions.

Synthesis and structure activity relationships (SAR) of RAR alpha-selective agonists are discussed. 4-[5-(5,8-Dimethyl-2H-3-chromenyl)-1H-2-pyrrolyl]benzoic acid (12a), which possesses a flat structural moiety and an oxygen atom at the hydrophobic part, showed highly selective transactivation activity at the RAR alpha receptor.

Daily intakes of 232Th and 238U and contributions of food categories to those nuclide intakes in Japanese were estimated using a market basket study for 18 food categories. Food categories having higher 238U contents (per g-fresh) were found to be as follows: seaweeds 1,140 microBq; fishes and shellfishes 37 microBq; nuts and seeds 11 microBq; bean products 8.6 microBq; and cooked meals 7.3 microBq. Big contributors to the daily 238U intake in Japan were as follows: seaweeds (50%); fishes and shellfishes (26%); and bean products (4.3%). For 232Th, higher contents (per g-fresh) were found as follows: seaweeds 28 microBq; fishes and shellfishes 13 microBq; nuts and seeds 8.2 microBq; green vegetables 3.9 microBq; cooked meals 3.5 microBq; and bean products 2.9 microBq. Big contributors to the daily 232Th intake were as follows: fishes and shellfishes (44%); green vegetables (11%); bean products (7.4%); and seaweeds (6.0%). For both nuclides, marine food products were big contributors, while minor contributors were oil and fats, eggs, and cooked meals. Daily intakes of 232Th and 238U in Japan were estimated to be 2.7 mBq and 14 mBq per person from the intakes of the 18 categories, respectively. Annual effective doses were estimated to be 232Th, 2.2 x 10(-7) Sv, and 238U, 2.2 x 10(-7) Sv.

Technetium-99 was determined in samples from the 30-km zone around the Chernobyl reactor. Concentrations of 99Tc in soil samples taken from three forest sites ranged from 1.1 to 14.1 Bq kg-1 dry weight for the organic soil layers, and from 0.13 to 0.83 Bq kg-1 dry weight for the mineral soil layers. In particular, for the organic layers, the measured 99Tc concentrations were one or two orders of magnitude higher than those due to global fallout 99Tc. The 99Tc depositions (Bq m-2), based on the sum of the depositions measured in organic and mineral layers, ranged from 130 Bq m-2 within the 10-km zone to about 20 Bq m-2 close to the border of the 30-km zone. Taking the corresponding measured 137Cs depositions into account, it was found that the activity ratio of 99Tc/137Cs ranged from 6 x 10(-5) to 1.2 x 10(-4). It was estimated that about 970 GBq of 99Tc had been released by the Chernobyl accident. This figure corresponded to 2%-3% of the total 99Tc inventory in the core.

Total diet samples were collected to estimate dietary intakes of several radionuclides for Ukrainian males by duplicate portion studies. The samples were collected in the Kiev, Rovno and Volynsky regions in autumn of 1994. Some foodstuffs, including milk and potato, were also collected in the same regions. Daily intakes of 232Th and 238U were estimated to be 2.0 mBq and 7.5 mBq per person, respectively. The estimated daily intake of 40K, 85 Bq per person, was equivalent to that of Reference Man (the intake of a West European and a North American). Daily intakes of 137Cs were found to be quite different between some regions. Daily intakes were 1.8 Bq per person in the Kiev region and 12 Bq per person in the Rovno and Volynsky regions. Daily intake of 134Cs was below 1 Bq per person.

This study was conducted to show a higher prevalence of interdigital tinea pedis in athletes by comparing athletes and non-athletes, and to examine swimming pools as a route of infection. The subjects were 282 athletes, 137 non-athletes, and 140 students enrolled in a swimming class at the University of Tsukuba. This study included the taking of cotton-swab samples from the interdigital skin surfaces of both feet and cultures, microscopical examinations of scales collected from subjects with lesions, and questionnaires. There was a significant difference between athletes and non-athletes in the prevalence of the relevant pathogens, and a higher risk of infection was shown in athletes. The study also found that 63.6% of the swimming class students were carriers, and that 85.0% of their dermatophytes were Trichophyton mentagrophytes. In addition, dermatophytes were also isolated from the floors of the swimming pool and the public baths. The results of these controlled studies suggested that there was a significant risk of dermatophytosis in both athletes and non-athletes using the swimming pool.

Prodigiosin 25-C and metacycloprodigiosin were found to suppress PTH-stimulated pit formation by cultured osteoclasts on bone slices. They also inhibited the acidification of vacuolar organelles in intact osteoclastic cells. Since the acidic pH in these organelles is generated by the action of proton-pumping ATPases of the organelle, these results indicate that the proton-pumping activity of V-ATPase in osteoclastic cells is essential in bone resorption and that the inhibition of the acidification of vacuolar organelles by prodigiosins results in suppression of PTH-stimulated bone resorption.

Analysis of global fallout 99Tc in environmental samples should provide useful information for predicting the nuclide behaviour under natural conditions which is important from the viewpoint of radioecology. Concentrations of 99Tc in rice paddy soils collected in Japan have been studied. After chemical separation, 99Tc in the final solution was measured by ICP-MS. The activity ratio of 99Tc/137Cs was used to understand the 99Tc behaviour in the environment because the fission yields of 99Tc and 137Cs from 235U or 239Pu are almost the same. The theoretical activity ratio from fission which is calculated now is about 3.0 x 10(-4). Our results showed that the range of activity ratios of 99Tc/137Cs in the soil samples was (2.0-5.2) x 10(-3); these ratios were one order of magnitude higher than the theoretical one. 99Tc has been accumulating in rice paddy soil like 137Cs has, although their mechanisms might differ. One of the reasons for the high ratio in the surface soil might be the ratios in the atmospheric samples, which have increased from the order of 10(-3) to 10(-2) (García-León et al., 1993).

As part of our research for the development of novel antiinflammatory drug candidates, we have designed and synthesized a series of (E)-3-(1,4-benzoquinonyl)-2-[(3-pyridyl)alkyl]-2-propenoic acid derivatives as dual inhibitors of 5-lipoxygenase (5-LO) and thromboxane (TX) A2 synthetase. In order to increase the absorption after oral administration, we introduced a carboxylic acid moiety into the 1,4-benzoquinone skeleton, which has 5-LO-inhibitory character. Introduction of a 3-pyridylalkyl group at the double bond of the 1,4-benzoquinonyl propenoic acid moiety afforded good to moderate inhibitory activities against the production of leukotriene (LT) B4 and TXA2 while not significantly inhibiting that of prostaglandin E2 by glycogen-induced peritoneal cells of rat (in vitro). The length of the methylene chain of the 3-pyridylalkyl group influenced the inhibition of LTB4 and TXB2 production. An increase of lipophilicity by introducing a more lipophilic alkoxy group did not markedly increase the inhibitory activity on LTB4 production. The position of alkoxy group on the 1,4-benzoquinone skeleton played an important role in TXA2 synthetase inhibition. Compounds such as 20c (E6700) with an appropriate alkoxy group and proper length of methylene side chain, together with a polar substituent (carboxylic acid), showed good inhibition of both 5-LO and TXA2 synthetase and possess a variety of pharmacologically beneficial effects.

The effects of the vacuolar-type H(+)-ATPase inhibitor bafilomycin A1 (baf.A1) and the carbonic anhydrase II inhibitor acetazolamide (AZ) on bone resorption and procathepsin L secretion of rat osteoclasts were investigated using the bone slice assay method, pit formation test. Baf.A1 completely suppressed osteoclastic bone resorption stimulated by parathyroid hormone (PTH), but did not affect procathepsin L secretion, while AZ suppressed both bone resorption and procathepsin L secretion. These findings suggest that bone resorption by procathepsin L secretion and its processing are regulated by proton production and proton secretion.

The suppressive effect of N-(benzyloxycarbonyl)-L-phenylalanyl-L-tyrosinal on bone resorption was examined in vitro and in vivo. This synthetic peptidyl aldehyde was found to be a potent and selective cathepsin L inhibitor in our screening for cysteine protease inhibitors. In the pit formation assay with unfractionated rat bone cells, 1.5 nM of this compound markedly inhibited parathyroid hormone-stimulated osteoclastic bone resorption. In addition, intraperitoneal administration of this peptidyl aldehyde (2.5-10 mg/kg) for 4 weeks suppressed bone weight loss dose dependently in the ovariectomized mouse, experimental model of osteoporosis. Hydroxyproline measurement of the decalcified femurs from these ovariectomized mice suggested that this compound acts as a bone resorption suppressor through the inhibition of collagen degradation.

Effects of concanamycin B, a specific inhibitor of the vacuolar type H(+)-ATPase (V-ATPase), on the stimulation of bone resorption induced by parathyroid hormone (PTH) were examined in vitro. Concanamycin B was found to inhibit PTH-stimulated osteoclastic pit formation and to suppress the acidification of vacuolar organelles by V-ATPase in the osteoclasts. PTH-stimulated 45Ca release from prelabelled chick embryonic calvariae was also inhibited by concanamycin B in a dose-dependent manner. These results suggest that osteoclastic acidification of lacunae by V-ATPase plays an essential role in mineral dissolution and degradation of the organic matrix during bone resorption.

The effects of short-term diet change from high fat (F) to high carbohydrate (C) (or vice versa) on the storage and utilization of glycogen and triacylglycerol (TG) in muscle and liver were studied in untrained rats. Rats were fed on an F or C diet for 28 days. For an additional 3 days, half of the rats in both F and C groups were fed the same diets as before (F-F and C-C) and the other half of the rats were switched to the counterpart diets (F-C and C-F). On the final day of the experiment, half of the rats in each diet group were exercised by swimming for 1.5 h and the other half were rested. Short-term diet change from F to C diets increased, but the change from C to F diets decreased, glycogen stores of soleus and plantaris muscles and liver, resulting in no difference in glycogen stores between F-C and C-C, and between F-F and C-F. The dietary change also had an affect on TG stores of red gastrocnemius muscle and liver-however, muscle TG stores were still higher in F-C than in C-C and C-F, and there were no differences in liver TG stores between F-C and C-F. Exercise decreased muscle glycogen contents markedly in F-C and C-C, whereas, it decreased muscle TG concentrations in F-F and C-F. Liver glycogen depletion was lower in F-C than in other groups. Lipolytic activities of epididymal adipose tissue at rest and postexercise were no differences between F-F and F-C, and were higher in F-C than in C-C and C-F. beta-adrenergic receptor binding was determined with [125I] iodocyanopindolol, and maximal numbers of beta-adrenergic receptor of plasma membrane from perirenal adipose tissue were approximately 170%-200% higher in F-C than in other groups at rest and postexercise. These results suggested that short-term C diet fed rats adapted to F diet enhanced not only glycogen stores of muscle and liver but also did not decrease lipolytic activity of adipose tissue with increased beta-adrenergic receptor density, resulting in the preservation of energy reserves (glycogen and TG) of muscle at rest, and liver glycogen sparing during exercise.

Secretion of procathepsin L into the culture medium from a bone cell mixture was markedly enhanced by addition of parathyroid hormone (PTH), 1 alpha,25-(OH)2D3 or tumor necrosis factor alpha (TNF alpha). These stimulators of secretion of procathepsin L enhanced bone pit formation, which was inhibited by E-64, but not by CA-074, a specific inhibitor of cathepsin B. Procathepsin L may thus participate in the process of bone collagenolysis during bone resorption. Procathepsin L partially purified from rat long bones under cold conditions was rapidly converted to the mature form under acidic conditions at room temperature. This conversion was inhibited by E-64, suggesting that the procathepsin L secreted into lacunae is catalytically converted to the mature enzyme by cysteine proteinase(s).

We have recently demonstrated that a rise in the extracellular divalent cation concentration induces a rapid elevation of cytosolic calcium in chick osteocytes. Here, we demonstrate that cytosolic calcium elevation that occurs in osteocytes on exposure to elevated extracellular calcium is independent of membrane voltage and is insensitive to modulation by organic calcium channel modulators, namely, BAY K 8644, nicardipine, and nifedipine. However, the calcium elevation was sensitive to modulation by an intracellular calcium antagonist, TMB-8, suggesting that the cytosolic calcium elevation was due to mobilization of this cation from an intracellular store.

Though it is proposed that osteocytes have several functions in bone metabolism, only limited definite information including results of calcium regulation has been forthcoming. We demonstrate here for the first time that the concentration of divalent cations, such as calcium, nickel and cadmium, directly regulates the cytosolic calcium concentration in osteocytes.

As part of our search for novel antiinflammatory drug candidates, we have designed and synthesized a series of 3-pyridylmethyl-substituted 2-amino-6- hydroxybenzothiazoles. Introduction of a 3-pyridylmethyl group into the 2-amino group (type-A) or the benzene ring (type-B) of 2-amino-6-hydroxybenzothiazoles imparted dual inhibitory activity against the production by glycogen-induced peritoneal cells of rat (in vitro) of leukotriene B4 (LTB4) and thromboxane A2 (TXA2), while not significantly inhibiting that of prostaglandin E2 (PGE2). The observed inhibition of the former two arachidonic acid metabolites was indicated to be the result of a direct action on 5-lipoxygenase and TXA2 synthetase by a cell-free in vitro assay. On the other hand, the inhibitory activities against PGE2 production were for most compounds very weak, indicating that they did not inhibit cyclooxygenase. Structure-activity relationship studies concerning the position of the 3-pyridylmethyl group revealed that type-B compounds generally showed about 10-fold stronger inhibitory activity against TXA2 synthetase than type-A compounds. The position of the 3-pyridylmethyl group played an important role in TXA2 synthetase inhibition. When some of these compounds (8, 13a, 26a (E3040), 26b, 27b, and 28b) were orally administered in the rat TNB/ethanol-induced chronic colitis model (100 mg/kg), the production of both LTB4 and TXB2 in the rat colon was reduced (ex vivo). In addition, one type-B compound, 6-hydroxy-5,7-dimethyl-2-(methylamino)-4-(3-pyridylmethyl)benzothiazole (26a), demonstrated a therapeutic effect at treatments of 100 mg/kg po once daily for 11 days and showed almost comparable activity to sulfasalazine at a dose of 500 mg/kg, the reference drug for inflammatory bowel diseases, in this in vivo model.

The secretion mechanisms of cathepsin L from osteoclasts in the process of bone resorption were investigated. The increases in bone pit numbers formed take place by PTH addition in parallel with the increases of cathepsin L and/or L-like proteinase activities in the culture medium of bone cells, and these were suppressed by the addition of calcitonin. The Z-Phe-Arg-MCA hydrolysing activity increased in the medium through the effect of PTH is considered to be a kind of procathepsin L by Western blotting analysis, and was suppressed by calcitonin addition. Furthermore, monensin inhibited not only the PTH-induced pit formation, but also cysteine proteinase activity in osteoclasts. Therefore, the procathepsin L excreted might be transferred from endothelial reticulum via Golgi and/or via lysosomes.

We have recently reported that rates of muscle glycogen repletion during the early period of recovery were increased by carbohydrate (CHO) loading in rats previously fed a high fat diet. However, the reason for this remained unanswered. The purpose of this study was to examine whether an increase of glycogen utilization due to an elevated pre-exercise glycogen store would enhance rates of glycogen repletion in muscle. Despite an equal degree of glycogen depletion, the rates of glycogen repletion of soleus, red and white gastrocnemius muscles by postexercise administration of glucose (3.0 g.kg-1 body mass) and citrate (0.5 g.kg-1 body mass) were faster in the CHO loaded (3 days) rats than in the nonloaded rats, as a result of elevated pre-exercise glycogen content and consequently the greater glycogen utilization. The higher rate of muscle glycogen repletion may in part be explained by increased postexercise glycogen synthase activity.

The proteinase responsible for bone collagen degradation in osteo-resorption was examined. The bone pit formation induced by parathyroid hormone (PTH) was markedly suppressed by leupeptin, E-64 and cystatin A, while no inhibition was observed by CA-074, a specific inhibitor of cathepsin B. Pig leucocyte cysteine proteinase inhibitor (PLCPI), a specific inhibitor of cathepsin L, and chymostatin, a selective inhibitor of cathepsin L, completely inhibited the pit formation. Cathepsin L activity in osteoclasts was much higher than the other cathepsin activities. Serum calcium in rats placed on a low calcium diet was decreased by treatment of E-64 or cystatin A, but not by CA-074. These findings suggest that cathepsin L is the main proteinase responsible for bone collagen degradation.

A new series of 3-(4-hydroxy-1-naphthalenyl)-2-propenoic acids was prepared and the inhibitory activities of its members on IL-1 generation were evaluated both by in vitro systems using human monocytes and/or rat exudated macrophages stimulated with LPS, and by an in vivo system using the rat CMC-LPS air-pouch model. Many compounds in this series were found to be potent inhibitors of IL-1 generation both in vitro and in vivo. Structure-activity relationships indicated that in the rat CMC-LPS air-pouch model by oral administration the (Z)-2-substituted propenoic acids with 3-alkoxy, 5-alkyl, and 4-hydroxy substituents on the naphthalene ring exhibit optimal inhibition. Among the compounds evaluated, (Z)-3-(5-ethyl-4-hydroxy-3-methoxy-1-naphthalenyl)-2-methyl-2-propeno ic acid (20a), which inhibited IL-1 generation from human monocytes with an IC50 value of 3.0 microM and had an IC50 value of 1.4 microM for rat exudated macrophages, showed the most potent inhibitory activity in the rat CMC-LPS model by oral administration. Compound 20a also showed antiinflammatory effects in animal models of inflammation.

Sixteen hybridomas secreting antibodies to HSV-1 and 22 hybridomas secreting antibodies to HSV-2 were derived from fusion of SP2/0 myeloma cells with spleen cells from BALB/c mice immunized with each respective virus. Four of the former 16 hybridomas and seven of the latter 22 hybridomas were subcloned and injected into pristane-primed mice to obtain high titers of monoclonal antibodies. Antigen specificity of these monoclonal antibodies were determined by the Western blotting (WB) assay. Two out of four monoclonal antibodies that showed selective reactivity for HSV-1 in IFA, reacted with HSV-1 specific proteins; #1 reacted with 100 KD and 70 KD proteins and #4 with a 150 KD protein, respectively, while the remaining two antibodies reacted only with a 50 KD protein that is type-common antigen. On the other hand, two out of seven antibodies which showed selective reactivity for HSV-2 in IFA, reacted with HSV-2 specific proteins: #5 with a 100 KD protein and #10 with three proteins of 30, 25, and 20 KD, and the other two antibodies reacted with a 50 KD protein that is a type-common antigen. The remaining three antibodies, two of which were found to be immunoglobulin type IgM, reacted with neither HSV-1 nor HSV-2 antigens in WB assay. In order to determine their utility in serotyping, 11 monoclonal antibodies were examined by IFA test for reactivity to cells that were infected with 20 HSV-1 or 16 HSV-2 isolates which had been typed by neutralization test.(ABSTRACT TRUNCATED AT 250 WORDS)

On the hypothesis that rat hepatic microsomal type I iodothyronine 5'-monodeiodinase (MDI) would be identical to hepatic protein disulfide isomerase (PDI) (Boado et al. Biochem. Biophys. Res. Commun. 155 1297-1304 (1988)), we examined the responses of these enzyme activities to dietary fat and thyroid status in rats. The hepatic MDI activity was higher in rats fed high fat diet than in rats fed low fat diet, but the hepatic PDI activity showed the reverse responses to the diets. Propylthiouracil administration (hypothyroidism) lowered the MDI activity, but elevated the PDI activity. Thyroxine administration (hyperthyroidism) elevated the MDI activity but had no effect on the PDI activity. These results indicate that the two enzyme activities are regulated by different mechanisms in vivo, suggesting that MDI and PDI are not identical enzymes.

Reported is a leiomyosarcoma of jejunum that was found in a 61-year-old man who was admitted to our clinic because of an abdominal distension. A CT scan and an ultrasonic examination revealed a giant tumor with a central necrosis in the abdominal cavity. A study of barium that was ingested revealed an elevated lesion without a fistel formation in the jejunum about 25 cm distant from the Treitz ligament. An ultrasonic the guided percutaneous biopsy was done and pathological diagnosis indicated a leiomyosaroma. Surgery was performed on June 16 1989 and the tumor weighing 4,934 g and 30 x 25 x 18 cm in size, was completely resected.