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Biomedical subjects

N Fujii

Publications and source records attributed to N Fujii.

At least 361 records · Page 20Linked to original sources
PubMed1992

Antibody response to oral streptococci in Behçet's disease.

The serum antibody titers against oral streptococci were studied by enzyme-linked immunosorbent assay (ELISA) both in patients with Behçet's disease (BD) and control groups. The patients with BD showed significantly higher antibody titers to S. sanguis strains 113-20, 114-23, and 118-1 which were isolated from patients with BD, in comparison with control groups. Also, the reactions of high-titered sera to the crude cell wall and soluble (or membrane) fractions of the 113-20 strain were observed by western blot test. The sera of the patients with BD demonstrated strong bands of approximately 36 kDa, 82 kDa, and 87 kDa in the crude cell wall fractions, and many bands of 80 kDa to 150 kDa in the membrane fractions, indicating that these proteins are the ones leading the high antibody titers to this bacterium in the sera of patients with BD.

Antibodies, Bacterial↗
PubMed1992

Caries experience in subjects 18-22 years of age after 13 years' discontinued water fluoridation in Okinawa.

The purpose of this study is to compare the state of teeth in young adults who had consumed fluoridated water from birth to 5-8 yr of age with the subjects who had non-fluoridated water. The sample consisted of 117 18-22-yr-old female nursing school students. The data obtained in dental examinations were sorted according to their place of residence, and then divided into groups depending on whether their place of residence had been supplied with fluoridated water or not. The fluoride concentration in the water had been 0.7-1.0 ppm in the six water supply systems which were under the control of the US military base. In the subjects who had fluoridated water, the following differences were found when compared to the control subjects: missing teeth and highly progressed caries occurred less frequently; significantly lower caries prevalence was present on free smooth and approximal surfaces; the overall DMFT differences in favor of the fluoridated subjects were small and non-significant.

Adolescent↗
PubMed1992

Induction of topoisomerase II-mediated DNA cleavage by the plant naphthoquinones plumbagin and shikonin.

Plumbagin and shikonin, plant metabolites which have naphthoquinone structures, induced mammalian topoisomerase II-mediated DNA cleavage in vitro. Treatment of a reaction mixture containing these naphthoquinones and topoisomerase II at an elevated temperature (65 degrees C) resulted in a great reduction in DNA cleavage, suggesting that the mechanism of the topoisomerase II-mediated DNA cleavage induced by these naphthoquinones is through formation of a cleavable complex, as seen with antitumor agents such as 4'-(9-acridinylamino)methanesulfon-m-anisidide and demethylepipodophyllotoxin ethylidene-beta-glucoside. Lawson and lapacol, which are structurally related plant metabolites with naphthoquinone moieties, could not induce topoisomerase II-mediated DNA cleavage. Plumbagin and shikonin induced a similar DNA cleavage pattern with topoisomerase II which was different from the cleavage patterns induced with other known topoisomerase II-active drugs. A DNA-unwinding assay with T4 DNA ligase showed that shikonin, lawson, and lapacol did not intercalate into DNA, while plumbagin and 2-methyl-1,4-naphthoquinone intercalate into DNA, but to a lower degree than 4'-(9-acridinylamino)methanesulfon-m-anisidide does.

Animals↗
PubMed1992

Anti-human immunodeficiency virus activity of a novel synthetic peptide, T22 ([Tyr-5,12, Lys-7]polyphemusin II): a possible inhibitor of virus-cell fusion.

More than 40 peptides associated with tachyplesin and polyphemusin, which are highly abundant in hemocyte debris of the horseshoe crabs Tachypleus tridentatus and Limulus polyphemus, were synthesized. Among these peptides, we found that a novel compound, which was called T22 ([Tyr-5,12, Lys-7]polyphemusin II), strongly inhibited the human immunodeficiency virus type 1 (HIV-1)-induced cytopathic effect and viral antigen expression. Its 50% effective concentration was 0.008 micrograms/ml, while its 50% cytotoxic concentration was 54 micrograms/ml. The anti-HIV activity of T22 was observed with several strains of HIV-1, including zidovudine-resistant strains, and with HIV-2 within the concentration range of 0.006 to 0.071 microgram/ml. T22 efficiently inhibited giant cell formation on the cocultivation of MOLT-4/HIV and MOLT-4 cells but modestly inhibited direct HIV binding. T22 did not inhibit reverse transcriptase activity. A time-of-addition study, which involved monitoring of the appearance of proviral DNA by using the polymerase chain reaction technique, found that T22 exerted its effect on a process, most probably virus-cell fusion or uncoating, immediately after virus adsorption.

Amino Acid Sequence↗
PubMed1992

Enzyme immunoassay of thyrotropin releasing hormone (TRH).

A sensitive and specific double-antibody enzyme immunoassay (EIA) for a thyrotropin releasing hormone (TRH)-like immunoreactive substance has been developed. In order to synthesize TRH-labeled beta-D-galactosidase (beta-gal), a newly devised TRH derivative, pGlu-His-Pro-NH-(CH2)6-NH2 (TRH-Hex), was employed. TRH-Hex was linked to beta-gal by the N-(epsilon-maleimidocaproyloxy) succinimide coupling procedure. For competitive reactions, the TRH antibody was incubated with standard TRH and TRH-Hex-beta-gal (delayed addition). Free and antibody-bound enzyme hapten were separated by using an anti-rabbit immunoglobulin G coated immunoplate. Activity of the enzyme on the plate was fluorometrically determined. The present immunoassay allows detection of 0.8 to 100 pmol/well of TRH.

Amino Acid Sequence↗
PubMed1992

An improved method for measurement of sweat expulsions during profuse sweating.

We present an improved ventilated-capsule method of recording for clear sweat expulsion patterns using nitrogen gas as a carrier gas heated to promote sweat evaporation. With this method, sweat expulsion patterns were more clearly recorded than with the conventional ventilated-capsule method. Taking the derivatives of these recordings of sweating expulsions could eliminate slow fluctuation components in the patterns of sweating. The results indicate that this method is useful in providing more-accurate measurements of sweat expulsion frequencies during profuse sweating.

Adult↗
PubMed1992

Radioactive cesium in dirt accumulations on the roof of buildings.

The concentrations of 137Cs and 134Cs in dirt deposits on the roofs of buildings are much higher than those in the surface of soil at ground level. Thus dirt on roofs concentrates radioactive cesium in fall-outs. The 137Cs concentration in dirt deposits on the roofs of older buildings is not consistently higher than that on the roofs of new ones, but the 137Cs/134Cs ratio is higher in deposits on older buildings constructed before the first half of the 1970s, and decreases exponentially with decrease in age of the buildings gradually reaching 1.9 +/- 0.2, the value in the air-borne dust at the time of the Chernobyl accident. From this relationship, the contribution of Chernobyl radioactivity to accumulated 137Cs was calculated as 32% on buildings constructed in 1962. The radioactive cesium concentrations in dirt deposits in gutters of private houses and on the roofs of university buildings in Japan were also determined.

Air Pollutants↗
PubMed1992

[A case of subacute traumatic aneurysm of the thoracic aorta with rare symptom of atelectasis in the left lung].

A case of subacute traumatic aneurysm of the thoracic aorta with atelectasis in the left lung is described. A 17-year-old male was admitted to our hospital because of traumatic shock. Radiographs revealed multiple fractures of upper and lower extremities, but on the chest roentgenogram there was no abnormal finding without slightly widened mediastinum. Next chest roentgenogram, four days later, showed left pleural effusion, atelectasis in the left lung and tracheal shift to the left. By the following examination of CT and DSA, the patient was diagnosed as a traumatic aneurysm at the aortic isthmus. The graft interposition of the descending aorta was performed with help of temporary bypass. He was making satisfactory progress after the operation. Reports about the operation of traumatic aneurysm of the thoracic aorta in subacute phase are relatively rare compared with those in chronic phase. No case, except this case, with tracheal shift to the left because of atelectasis in the left lung has been reported previously in Japan. It must be considered that the trachea may be shifted to the left by atelectasis of the left lung like this case.

Adolescent↗
PubMed1991

Interactions of an antimicrobial peptide, tachyplesin I, with lipid membranes.

Tachyplesin I, isolated from the acid extracts of hemocytes of Tachypleus tridentatus, is a cyclic broad-spectrum antimicrobial peptide forming a rigid, antiparallel beta-sheet because of two intramolecular S-S linkages. The strong binding of the peptide to lipopolysaccharides cannot explain the susceptibilities of gram positive bacteria and fungi to the peptide. We found that tachyplesin I caused a rapid K+ efflux from Escherichia coli cells, concomitant with a reduced cell viability. This result suggests that the peptide-induced permeability enhancement of the bacterial membranes may be a plausible action mechanism. Thus, we studied the interactions of tachyplesin I with various large unilamellar vesicles (LUVs) to reveal the molecular machinery of the antimicrobial activity. Tachyplesin I induced the leakage of calcein, a trapped fluorescent marker, from LUVs of acidic phospholipids, especially phosphatidylglycerol (PG), but not from phosphatidylcholine LUVs. A detailed analysis found that the affinity of the peptide to the PG membranes is very strong and that the binding of one peptide molecule to approx. 200 lipid molecules leads to a significant leakage. The location of tachyplesin I in membranes was estimated by use of the Trp-2 fluorescence of the peptide. The presence of PG LUVs caused a blue shift of the maximum wavelength, an increase in the quantum yield, and a complete protection from fluorescence quenching by an aqueous quencher, acrylamide. Moreover, the degree of fluorescence quenching of the Trp residue by n-doxylstearates was in the order n = 5 greater than 7 greater than 12 approximately equal to 16. These results show that the Trp residue of tachyplesin I seems to locate in a hydrophobic environment near the surface of the PG bilayers.

Amino Acid Sequence↗
PubMed1991

Gastrin releasing peptide-like immunoreactive substance in rat mammary glands during pregnancy.

The concentration of gastrin releasing peptide-like immunoreactive substance (GRP-IS) was measured by enzyme immunoassay (EIA) in rat mammary glands during pregnancy and after delivery. The GRP-IS concentration was high in the middle stage of pregnancy (10-14 days of gestation) and decreased in late pregnancy to reach a plateau range. By using HPLC, it was shown that GRP(20-29) and GRP(16-29) were mainly present in rat mammary glands. Immunohistochemical study revealed that epithelial cells of rat mammary glands were stained intensely with antiserum GP-6201. These results suggest that GRP-IS is produced and secreted in epithelial cells of mammary glands and takes part in the proliferation, differentiation and hypertrophy of mammary glands.

Animals↗
PubMed1991

Purification and amino acid sequence of a bitter gourd inhibitor against an acidic amino acid-specific endopeptidase of Streptomyces griseus.

An inhibitor (BGIA) against an acidic amino acid-specific endopeptidase of Streptomyces griseus (Glu S. griseus protease) was isolated from seeds of the bitter gourd Momordica charantia L., and its amino acid sequence was determined. The molecular weight of BGIA based on the amino acid sequence was calculated to be 7419. BGIA competitively inhibited Glu S. griseus protease with an inhibition constant (Ki) of 70 nM, and gel filtration analyses suggested that BGIA forms a 1:1 complex with this protease. However, two other acidic amino acid-specific endopeptidases, protease V8 from Staphylococcus aureus and Bacillus subtilis proteinase (Glu B. subtilis protease), were not inhibited by BGIA. BGIA had no inhibitory activity against chymotrypsin, trypsin, porcine pancreatic elastase, and papain, although subtilisin Carlsberg was strongly inhibited. The amino acid sequence of BGIA shows similarity to potato chymotrypsin inhibitor, barley subtilisin-chymotrypsin inhibitor CI-1 and CI-2, and leech eglin C, especially around the reactive site. Although the residue at the putative reactive site of these inhibitors is leucine or methionine, the corresponding amino acid in BGIA is alanine.

Amino Acid Sequence↗
PubMed1991

Chemoselective one-step purification method for peptides synthesized by the solid-phase technique.

The specific reaction between SH and iodoacetamide groups has been explored as the basis of an affinity-type purification procedure for peptides synthesized by the solid-phase technique. For this affinity-type purification procedure, we synthesized an SH precursor reagent bearing an acid-labile S-protecting group, pMB-SCH2CONHCH2CH2-SO2CH2CH2OCO2pNP (compound I), in which pMB is p-methoxybenzyl and pNP is p-nitrophenyl. Using this reagent, the procedure involves the following sequence of four reactions: (i) attachment of the SH function of compound I to the alpha-amino group of a peptide-resin through a base-labile sulfonylethoxycarbonyl linkage in the final step of solid-phase peptide synthesis, (ii) acid treatment to remove the S-pMB and side-chain-protecting groups employed and cleave the modified peptide from the resin, (iii) immobilization of the derived SH-peptide on an iodoacetamide-resin column, and (iv) base (5% NH4OH) treatment to release the desired peptide from the resin in nearly pure form. To facilitate this purification procedure, unreacted amino groups were acetylated in each step during solid-phase synthesis. The usefulness of this method was demonstrated by the purification of several peptides (18 to approximately 44 amino acids in length) synthesized by the 9-fluorenylmethoxycarbonyl (Fmoc)-based solid-phase technique. The principle of this affinity-type purification procedure may also be applied to the tert-butoxycarbonyl (Boc)-based solid-phase technique.

Chromatography, High Pressure Liquid↗
PubMed1991

Induction of mammalian DNA topoisomerase I and II mediated DNA cleavage by saintopin, a new antitumor agent from fungus.

Saintopin is an antitumor antibiotic recently discovered in mechanistically oriented screening using purified calf thymus DNA topoisomerases. Saintopin induced topoisomerase I mediated DNA cleavage comparable to that of camptothecin, and topoisomerase II mediated DNA cleavage equipotent to those of 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA) or 4'-demethylepipodophyllotoxin 9-(4,6-O-ethylidene-beta-D-glucopyranoside) (VP-16). Treatment of a reaction mixture containing saintopin and topoisomerase I or II with either elevated temperature (65 degrees C) or higher salt concentration (0.5 M NaCl) resulted in a substantial reduction in DNA cleavage, suggesting that the topoisomerase I and II mediated DNA cleavage induced by saintopin is through the mechanism of stabilizing the reversible enzyme-DNA "cleavable complex". Consistent with the cleavable complex formation with both topoisomerases, saintopin inhibited catalytic activities of both topoisomerase I and topoisomerase II. The DNA cleavage intensity pattern induced by saintopin with topoisomerase I was different from that by camptothecin. A difference in cleavage pattern was also detected between saintopin and m-AMSA or VP-16 in topoisomerase II mediated DNA cleavage. DNA unwinding assay using T4 DNA ligase showed that saintopin is a weak DNA intercalator like m-AMSA. Thus, saintopin represents a new class of antitumor agent that can induce both mammalian DNA topoisomerase I and mammalian DNA topisomerase II mediated DNA cleavage.

Amsacrine↗
PubMed1991

Induction of a heat-stable topoisomerase II-DNA cleavable complex by nonintercalative terpenoides, terpentecin and clerocidin.

Terpentecin and clerocidin, microbial terpenoides, have been known to be potent antitumor antibiotics. However, the critical biochemical target of these terpenoides has not been identified. Our present studies, using purified mammalian topoisomerase II, have shown that terpentecin and clerocidin induce topoisomerase II-mediated DNA cleavage in vitro with comparable potency to that of demethylepipodophyllotoxin ethylidene-beta-D-glucoside. These terpenoides produced a similar DNA cleavage pattern which is distinctly different from those generated in the presence of the known topoisomerase poisons, demethylepipodophyllotoxin ethylidene-beta-D-glucoside and 4'-(9-acridinylamino)methanesulfon-m-anisidide. Brief heating at 65 degrees C, which abolishes completely the cleavable complex with demethylepipodophyllotoxin ethylidene-beta-D-glucoside, of the reaction mixture containing these terpenoides resulted in slight reduction in DNA cleavage. Thus, differently from other topoisomerase II-active antitumor agents, terpentecin and clerocidin induce formation of a cleavable complex which is stable for heat or salt treatments. The lack of significant DNA binding or intercalation activity of terpentecin and clerocidin suggests that topoisomerase II is a cellular target for these drugs.

Anti-Bacterial Agents↗
PubMed1991

Further characterization of the autologous mixed lymphocyte reaction: induction of double negative gamma delta T lymphocytes.

To investigate the specific nature of the autologous mixed lymphocyte reaction (AMLR), we applied a method in which mixtures of NY-nonadherent responder cells and NY-adherent stimulator cells were treated with neuraminidase before culture and then cultured to assay the AMLR. This method produced a marked enhancement of DNA replication in the responder cells and the results were reproducible, regardless of the individuals tested. Using this method, we were able to make the following observations regarding the specific nature of the AMLR. (i) The AMLR is an IL-2-independent reaction, as revealed by bioassay to detect the presence of IL-2 by a blocking test using anti-IL-2R sera and as shown by the absence of mRNA for IL-2 in Northern hybridization. (ii) It is also HLA-DR dependent as proven by the fact that anti-DR sera almost completely inhibited the reaction. (iii) The AMLR was also found to induce the generation of activated CD4+ helper T cells in direct response to stimulation by NY-adherent cells, in which HLA-DR antigens were involved. (iv) Also, it induced the generation of CD4-CD8- double-negative (DN) lymphocytes, including gamma delta T cells with a cytotoxic activity against NK-resistant target cells and with a variety of lymphocyte activation markers (CD56, HLA-DR, CD25, transferrin receptors, CD38, and LFA-1). However, the AMLR did not induce the generation of NK cell markers CD16 and CD57. (v) The DN lymphocytes and gamma delta T cells appeared to be generated from the precursors of CD4-CD8- DN cells, in direct response to the stimulator cells. These results strongly suggest that the AMLR may be a phenomenon which induces the proliferative response of gamma delta T cells and their precursors, in addition to that of alpha beta T cells, particularly of CD4+ helper T cells.

Adult↗
PubMed1991

Physicochemical determinants for the interactions of magainins 1 and 2 with acidic lipid bilayers.

Permeability enhancement of acidic lipid small unilamellar vesicles (dioleoylphosphatidylglycerol, DOPG; dipalmitoylphosphatidylglycerol, DPPG; bovine brain phosphatidylserine, PS) induced by magainins 1 and 2, basic antimicrobial peptides from Xenopus skin, was investigated at 30 degrees C based on leakage of calcein, an entrapped fluorescent marker. Both the peptide concentration and the lipid concentration dependencies of the leakage rate were analyzed to obtain the binding isotherms of the peptides to the membranes and the 'membrane-perturbing activities' of the membrane-bound peptides. For both peptides, the binding affinity was in the order DOPG greater than DPPG greater than PS, which coincided with the zeta potential order (-54, -39, and -9 mV, respectively). An increase in salt concentration of the medium reduced binding and leakage. Electrostatic interactions play a crucial role in the binding process. On the other hand, the membrane-perturbing activity is regulated by membrane fluidity: The fluid membranes (DOPG and PS) were leakier. A circular dichroism study suggested that at least 14 positively charged residues in the N-terminal regions can form amphiphilic helices which interact with the membranes. An even stronger binding of magainin 2 can be explained in terms of more positive charges in its N-terminal region. A tentative model for the magainin-lipid interactions is hypothesized.

Amino Acid Sequence↗