Mutagen formation and nitration by exposure of phenylphenols to sunlight in water containing nitrate or nitrite ion.
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Biomedical subjects
Publications and source records attributed to A Ito.
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Forty-six sclerotherapy sessions were performed on liver cirrhotics with high-risk esophageal varices using GT XIII, a sclerosant composed of gelatin, thrombin and coagulation factor XIII. GT XIII was effective for the prevention of temporary symptoms and transient hypotension observed in 55 sclerotherapy sessions using thrombin. In 42 (91%) sessions, patients underwent sclerotherapy with no symptoms, and in the other four (9%) sessions, only slight symptoms of general fatigue and headache were observed. Changes in the mean arterial pressure were significantly smaller in sessions using GT XIII than in those using thrombin (-12.3 +/- 13.6 vs. -26.8 +/- 20.7 mmHg, P less than 0.01). Changes in coagulation tests, similar to those of disseminated intravascular coagulation (DIC), were also reduced in sessions using GT XIII. Urinary kallikrein and kinin excretion significantly increased after the procedure (P less than 0.01), indicating activation of the renal kallikrein-kinin system. Increases in urinary kallikrein and kinin excretion showed a significant relationship with the consumed plasma fibrinogen levels (r = -0.51, P less than 0.01 and r = -0.58, P less than 0.01, respectively), and it was suggested that activation of the glandular kallikrein-kinin system caused by abrupt DIC-like changes in the hemostatic system might play a role in manifestations of temporary complications occurring with the use of hemostatic agents containing thrombin.
The potential modifying effects of high prolactinemia on rat prostate carcinogenesis was investigated. Male F344 rats were treated at 5 times of 5-week intervals with s.c. injections of 3,2'-dimethyl-4-aminobiphenyl (DMAB), each injection following 3 weeks pretreatment with dietary ethinyl estradiol. After completion of the carcinogen administration stage, rats received multiple s.c. transplantations of a prolactin producing transplantable pituitary tumor, MtT/F84 until sacrifice at week 51. The effects of additional or single treatment with bromocriptine, a prolactin suppressing agent, were also investigated. The body, liver and kidney but prostate weights were significantly increased in the groups given MtT/F84. Although the development of prostate carcinomas was not affected by the observed hyperprolactinemia, the incidences of atypical hyperplasia of both ventral and lateral prostate were significantly enhanced. The findings thus indicate that prolactin may have promoting potential for prostate carcinogenesis.
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The distribution of the alpha subspecies of protein kinase C (PKC) in rat brain was demonstrated immunocytochemically by using polyclonal antibodies raised against a synthetic oligopeptide corresponding to the carboxyl-terminal sequence of alpha-PKC. The alpha-PKC-specific immunoreactivity was widely but discretely distributed in both gray and white matter. The immunoreactivity was associated predominantly with neurons, particularly with perikaryon, dendrite, or axon, but little was seen in the nucleus. Glial cells expressed this PKC subspecies poorly, if at all. The highest density of immunoreactivity was seen in the olfactory bulb, septohippocampal nucleus, indusium griseum, islands of Calleja, intermediate part of the lateral septal nucleus, and Ammon's horn. A moderately high density of the immunoreactivity was seen in the anterior olfactory nucleus, anterior commissure, cingulate cortex, dentate gyrus, compact part of the substantia nigra, interpeduncular nucleus, inferior olive, and olivocerebellar tract. This distribution pattern was consistent with that obtained by in situ hybridization histochemistry. The distribution of alpha-PKC immunoreactivity was different from that of beta I-, beta II-, and gamma-PKC immunoreactivity. These findings suggest that alpha-PKC is involved heavily in the control of specific functions of some restricted neurons.
Investigations of trombiculid mites and Rickettsia tsutsugamushi in wild rodents were made in southern Gifu Prefecture where patients infected with tsutsugamushi disease recently have been found. A total of 16,396 trombiculid mites, consisting of 10 species from three genera, was collected from 170 Apodemus speciosus in two locations. Kani-Sakahogi and Kuze. Leptotrombidium scutellare (Nagayo et al.) (44.0%) was most predominant, followed by L. pallidum (Nagayo et al.) (26.9%); L. fuji (Kuwata et al.) (13.6%); and Gahrliepia saduski Womersley (14.2%). These four species constituted the bulk of the chigger mite fauna. L. scutellare was present from October to February with a remarkably high peak in November, whereas pallidum occurred from November to March with the highest peak in December. L. fuji and G. saduski showed their highest peaks in December and moderate peaks in early summer (April and May). Positive identification of Rickettsia tsutsugamushi in wild rodents from Kani-Sakahogi were found to be 50 and 58.3% in November 1985 and 1986, respectively, and 38.5% in November 1986 from Kuze. R. tsutsugamushi was isolated from chigger mites of an L. pallidum-rich group, displaying the highest titer to Karp strain. Serological investigation of rodents to R. tsutsugamushi antibodies were calculated as 41.6 and 50% positive in November 1985 and 1986 in Kani-Sakahogi, respectively, and 50% in November 1986 in Kuze. The Karp strain was dominant in specificity to antibodies. These results indicate that the surveyed areas have a high probability of occurrence of tsutsugamushi disease, and L. scutellare and L. pallidum may serve as the vectors in these areas. Particularly, we suggest that L. scutellare is the most important vector which has caused a recent outbreak of this disease in southern Gifu Prefecture.
Seeds of Wisteria floribunda contain several kinds of cysteine proteinase inhibitor (cystatin). We purified and characterized one of these inhibitors, named WCPI-3. The molecular weight of WCPI-3 was estimated to be 17,500 and 15,700 by gel filtration and SDS-PAGE, respectively. The isoelectric point was 5.7. WCPI-3 formed an equimolar complex with native papain and the dissociation constant was estimated to be 6.1 nM. Complex formation between WCPI-3 and Cys25-modified papain, such as S-carboxy-methylated or S-carbamoylmethylated papain, could not be observed by gel filtration or native PAGE analysis. A peptide fragment derived from WCPI-3 digested by Achromobacter proteinase (lysyl endopeptidase) had the amino acid sequence of VVAGVNYRFVLK. The VVAG sequence in this fragment corresponds to the conserved sequence QVVAG which is considered to be one of binding regions to cysteine proteinases. The amino acid sequence of the amino-terminal portion (34 residues) of WCPI-3 was highly homologous to that of oryzacystatin from rice seeds.
An acidic amino acid-specific endopeptidase was purified from Protease Type XVI (Sigma), a commercial product from culture filtrate of Bacillus subtilis, by a series of column chromatographies on CM-Toyopearl (Fractogel) and Mono-S, guided by activity assay using Boc-Ala-Ala-Pro-Glu-pNA as a substrate. The final preparation was homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and reversed-phase HPLC. The molecular weight of the protease was estimated to be 18,000 by gel filtration on TSK gel G3000SWXL column using 6 M guanidine hydrochloride as an eluent, and 17,000 by SDS-PAGE in the presence of 2-mercaptoethanol. The isoelectric point of the protease was 7.7. Studies on the substrate specificity with peptide p-nitroanilides and natural peptides revealed that the protease hydrolyzes the peptide bonds on the carboxyl-terminal side of acidic amino acids, especially of glutamic acid. The protease was completely inactivated by DFP, indicating the serine protease nature of the protease. The activity of the protease was also inhibited by EDTA and GEDTA, and reactivated by Ca2+. The protease contained 1.3 +/- 0.2 mol/mol protein of Ca2+. These results suggest that Ca2+ plays a vital role in the protease activity.
The efficacy of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on neutropenia was evaluated in 14 patients with AIDS and AIDS-related complex (ARC). In all patients, including 11 neutropenic patients, 100 or 200 micrograms/m2 of rhG-CSF significantly increased the neutrophil counts. The response was greater in patients with higher neutrophil counts before the treatment, and was also dose-dependent. Although the effect seemed to be less potent, the agent also increased the neutrophil counts even when zidovudine (azidothymidine, AZT) and other myelosuppressive antiviral agents were administered simultaneously. These observations indicate that rhG-CSF may be beneficial in preventing and treating some secondary infections, and will make it easier to continue therapy with antiviral agents in patients with AIDS or ARC.
MtT/F84 grew well in Fischer rats (F344), but tumor growth was promoted in hyperestrogenized rats. Effects of dietary retinoic acid (RA) on tumor growth, estrogen receptor (ER) and serum growth hormone (GH) level were examined. Tumor latency became shortened, and tumor take and weight were promoted by all-trans RA at dosages of 50 and 200 mg/kg basal diet, but not dose-dependently. ER level was elevated in tumor of RA-treated rats, whereas the retinoic acid-binding protein level remained unchanged. RA also elevated incorporation of 5-bromo-2'-deoxyuridine, a thymidine analogue, into DNA of tumor cells. Average serum GH level was increased in tumor-bearing rats treated with RA and was well correlated with tumor weight. RA may directly affect ER level and enhance estrogenic action, resulting in promotion of tumor growth, or it may act independently for tumor growth and elevation of serum GH level.
Production of the elastase-like enzyme, Suc-(Ala)3-p-nitroanilide hydrolase, was examined in human uterine cervical fibroblasts. The metallo-dependent enzyme is present in both media and cellular fractions. Human recombinant interleukin-1 alpha stimulated cells to produce and secrete this elastase-like enzyme. These results suggest that modulation of this enzyme activity by interleukin-1 may play a significant role in the ripening and dilation of the cervix, because elastin is a component of the uterine cervix and its peptides are known to be chemotactic for leukocytes.
Four kinds of cultured clonal cell line were established from estrogen-induced mammotropic pituitary tumor. These clones showed differences of hormone secretion, morphology, and response to estrogen. One clone tentatively designated MtT/E, consisted of spindle-shaped epithelial cells and showed the strongest adhesion to the culture dish, but did not secrete any pituitary hormone. The second cell line, designated MtT/S, secreted only GH, showed very weak contact with the culture dish, and proliferated almost anchorage independently. the MtT/S cells were mainly spherical and formed floating or weakly adherent clusters. Some of them had very long dendrite- or neurite-like cell processes. Electron microscopic examination of the MtT/S cells showed a normal somatotrope-like appearance, i.e. the presence in the cytoplasm of many GH-containing secretory granules, well developed rough endoplasmic reticulum, and Golgi apparatus. Furthermore, these cells secreted GH in response to stimulation with GRF. The third cell line with an ovoid cell appearance tentatively designated MtT/SM, secreted both PRL and GH, and showed anchorage-dependent proliferation. The fourth cell line designated MtT/Se secreted only a small amount of GH. Among these newly established cell lines, MtT/Se was the smallest in size and showed estrogen-dependent proliferation. The many small secretory-like granules present in the cytoplasm of MtT/Se cells showed no immunocytochemically positive reaction for anterior pituitary hormone. MtT/S and MtT/SM cell lines were also sensitive to estrogen.
The cDNA for rat monoamine oxidase A (MAO-A) mRNA was isolated from a liver cDNA library in lambda gt11 as the cDNA species cross-hybridizing with rat monoamine oxidase B (MAO-B) cDNA. The primary structure of the protein, deduced from the nucleotide sequence, consisted of 526 amino acid residues and its molecular mass was calculated to be 59.6 kD. The amino acid sequence shows about 70% identity with that of rat MAO-B. Northern blot analysis showed that MAO-B mRNA was expressed in various rat tissues and the highest expression was observed in heart. The MAO-B mRNA was quite different from MAO-A mRNA in the tissue-specific expression and liver had the highest level of the mRNA.
The compounds with efficient DNA transfection ability into eukaryotic cells were searched from various synthetic amphiphiles which have cationic heads and long saturated hydrocarbon tails. The efficiency of amphiphiles in gene transfer was examined by the transient expression of cytochrome b5 from its cDNA in COS cells. Among various synthetic amphiphiles, including N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride which is commercially available lipid, O,O'-didodecyl-N-[p-(2-trimethylammonioethyloxy)benzoyl]-(L) -glutamate bromide was highest in efficiency. The optimum condition for the amount of the amphiphile and DNA, and the incubation time were established to be 7.5-15 micrograms/22 mm dish and 1-10 micrograms/22 mm dish, and 48-72 h, respectively.
Pain and detection thresholds upon Nd:YAG laser irradiation were studied in healthy human subjects. The output power was set at 100 mW (1.36 W/cm2), 300 mW (4.11 W/cm2), 600 mW (8.22 W/cm2), 800 mW (11.0 W/cm2) and 1,000 mW (13.7 W/cm2). The duration was 5 seconds. The irradiation was done to the hand dorsum using the non-contact method (distance between the subject and the laser tip was 5.0 mm). After irradiation, the detection threshold was about 600 mW and the pain threshold was about 1,000 mW. The skin temperature was 34.9 +/- 0.64 degrees C before irradiation and 38.0 +/- 0.45 degrees C after a 5-sec-irradiation at 1,000 mW. No skin damage was observed after the irradiations.
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The effect of a novel nonsteroidal anti-inflammatory drug, 2-(10,11-dihydro-10-oxo-dibenzo[b,f]thiepin-2-yl)propionic acid (CN-100), on the biosynthesis of proteoglycans in chondrocytes of rat femoral head articular cartilage was studied. Sodium salicylate and hydrocortisone inhibited significantly the biosynthesis proteoglycans, but CN-100 did not affect the biosynthesis of glycosaminoglycan chains nor their sulfation. The effect of CN-100 on the interleukin 1-induced biosynthesis of prostaglandin E2 (PGE2) and tissue collagenase in rabbit and human rheumatoid synovial fibroblasts was also examined. CN-100 did not inhibited directly the collagenase production, but significantly prevent the biosynthesis of PGE2 known as an inflammatory mediator. The 50% inhibitory concentration (IC50) of CN-100 on PGE2 of rabbit and human fibroblasts were 3.8 x 10(-9) M and 2.4 x 10(-8) M, respectively; an inhibitory effect of CN-100 was equivalent to that of a control drug, hydrocortisone (2.0 x 10(-9) M and 1.7 x 10(-8) M, respectively). These results indicate that CN-100 is a potent anti-inflammatory drug without affecting the proteoglycan biosynthesis.