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Biomedical subjects

K Fujikawa

Publications and source records attributed to K Fujikawa.

At least 199 records · Page 11Linked to original sources
PubMed1990

Placental anticoagulant protein-I: measurement in extracellular fluids and cells of the hemostatic system.

Placental anticoagulant protein-I (PAP-I), a member of the lipocortin protein family, is a potent in vitro anticoagulant whose in vivo function is unknown. Very low levels of PAP-I were present in plasma of normal volunteers (0 to 5 ng/ml) and in randomly chosen plasma specimens from hospitalized patients (0 to 28 ng/ml). Review of selected hospital records did not reveal any single clinical entity that correlated with plasma levels. PAP-I was also found in amniotic fluid (12 to 107 ng/ml) and in conditioned medium of cultured endothelial cells (49 +/- 20 ng/ml). Gel filtration experiments showed that PAP-I was intact and uncomplexed in plasma and amniotic fluid. The protein was fairly abundant intracellularly: 4080 +/- 2560 ng/mg total protein in cultured umbilical vein endothelial cells; 178 +/- 109 ng/mg in platelets; 564 +/- 384 ng/mg in leukocytes; and 8.4 +/- 4.3 ng/mg in erythrocytes. The levels of PAP-I increased in platelet-rich plasma after stimulation of platelets with arachidonic acid but not after stimulation with ADP, epinephrine, thrombin, ristocetin, or collagen. These data suggest that PAP-I probably does not function as a circulating natural anticoagulant in normal persons.

Amniotic Fluid↗
PubMed1990

[Xenon enhanced dynamic computed tomography of the lung; regional ventilation measurement].

CT-functional images (CT-FI's) were created from Xe-dynamic CT data of 10 patients with chronic pulmonary emphysema and 6 volunteers with normal lung. In CT-FI's of normal lungs, the values of washin and washout parameter of outer zone were larger than those of inner zone. In CT-FI's of chronic pulmonary emphysema, the values of washin parameter of inner zone were larger than those of outer zone and the value of washout parameter of outer zone were larger than those of inner zone. CT-FI's proved to helpful in evaluating the axial distribution of ventilation dynamics in chronic pulmonary emphysema.

Administration, Inhalation↗
PubMed1989

Phospholipid binding properties of human placental anticoagulant protein-I, a member of the lipocortin family.

Human placental anticoagulant protein-I (PAP-I) is a member of the lipocortin/calpactin/annexin family of Ca2+-dependent phospholipid binding proteins. PAP-I was labeled with fluorescein 5-isothiocyanate (1 mol/mol); this derivative had anticoagulant activity identical to the unlabeled protein and could be used to measure Ca2+-dependent binding to phospholipid vesicles through changes in fluorescence quenching. At 1.2 mM Ca2+, 0.50 M ionic strength, pH 7.4, 25 degrees C, fluorescein-labeled PAP-I bound to phospholipid vesicles containing 80% phosphatidylcholine, 20% phosphatidylserine with a Kd of 1.2 +/- 0.2 nM (mean +/- S.D.). At an ionic strength of 0.15 M, the Kd decreased to less than 0.1 nM. Prothrombin and factor Xa both competed with fluorescein-labeled PAP-I for binding to anionic phospholipid vesicles, but with affinities at least 1000-fold weaker than PAP-I. PAP-I bound only weakly (Kd greater than 2 x 10(-5) M) to neutral or anionic phospholipid monomers, and this binding was not calcium-dependent. These results show that the affinity of PAP-I for anionic phospholipid surfaces is sufficient to explain its potency as an in vitro anticoagulant.

Annexins↗
PubMed1989

Primary structure of a protein C activator from Agkistrodon contortrix contortrix venom.

The amino acid sequence of a protease, protein C activator, from Agkistrodon contortrix contortrix venom was determined. Peptide fragments obtained by chemical or enzymatic cleavage of the S-carboxymethylated protein were purified by gel filtration and reverse-phase high-performance liquid chromatography. The present study demonstrates that protein C activator from A. contortrix contortrix venom is a trypsin-type serine protease that is composed of 231 residues with a molecular weight of 25,095 for the polypeptide portion of the molecule. By analogy to the mammalian serine proteases, the catalytic triad in venom protein C activator consists of His-40, Asp-85, and Ser-177. The protein also contains three N-linked glycosylation sites at Asn-21, Asn-78, and Asn-129. The amino acid sequence of protein C activator exhibits a high degree of sequence identity with other snake venom proteases: 73% with batroxobin, 68% with flavoxobin, and 55% with Russell's viper venom factor V activator.

Amino Acid Sequence↗
PubMed1989

Arthroscopy after anterior cruciate reconstruction with the Leeds-Keio ligament.

The healing of anterior cruciate ligaments reconstructed with the Leeds-Keio artificial ligament was observed by arthroscopy in 42 knees and biopsy in 19 knees at intervals from 3 to 24 months after implantation. By three months the implant was covered with immature new tissue, and a dense vascular network crossed its surface. At 12 months a new ligament had developed and matured, looking like the natural one in most cases. Histology at this stage showed abundant collagenous fibres running parallel and longitudinally, while the synovial membrane showed no more than very slight inflammatory changes. By 18 to 24 months, the new ligament often had the arthroscopic appearance of a normal anterior cruciate ligament. These results suggest that this scaffold type of artificial ligament is effective for cruciate reconstruction, giving satisfactory healing without significant complications.

Adolescent↗
PubMed1989

[Clinical application of tetracycline-containing strips (LSD) in periodontal disease].

The purpose of this study was to evaluate the clinical application of strips containing 10% Tetracycline (TC) in periodontal disease. In this double blind study, placebo strips, which did not contain TC, were also prepared. Ninety-two sites in 46 cases with greater than a 4 mm periodontal pocket were randomly assigned to experimental and control study groups. The following clinical parameters at each site were recorded at Weeks 0, 1, 2 and 3 after insertion of the LSD or placebo strip: plaque and calculus score, probing depth, gingival inflammation (redness, swelling, bleeding on probing, pus discharge), mobility of the tooth, pain on insertion of strip and pain after the procedure. The residual quantity of TC in the periodontal pocket was also determined at 1, 2 and 7 days after inserting the LSD. All clinical data was evaluated using 3 types of statistical test (X2 test, Mann-Whitney's U test and Wilcoxon's test). Thus, the following results were obtained: 1. The LSD group showed much improvement in clinical condition compared with the placebo group. 2. The conditions of bleeding, redness or swelling of gingival tissue were much improved in the LSD group, even after data evaluation with all 3 types of statistical test. 3. The residual quantity of TC was decreased day by day until finally it was not recognized in the 7 day samples. 4. This study suggested that the application of LSD in the periodontal pocket could be effective in periodontal treatment.

Humans↗
PubMed1989

[pH determination in human crevicular fluids. Examination of the pH meter and evaluation of the correlation between pH level and clinical findings or the microflora in each periodontal pocket].

The purpose of this study was to investigate the accuracy and reproduction of a microelectrode with a glass capillary, and to discuss the correlation between the pH level and the microflora in periodontal pockets or the clinical findings in the various stages of periodontal disease. The upper or lower anterior teeth of 34 patients with mild to severe periodontal disease were examined. Before the measurement, supragingival plaque was removed from each tooth. A special glass capillary was incorporated in the microelectrode to cover its tip, so that it would not touch the pocket walls or tooth surfaces during pH measurement. After the pH in the crevicular fluid was determined, subgingival plaque was collected with paper points from each periodontal pocket. This bacterial plaque was sonicated with saline solution, and examined under a phase contrast microscope with a calibrated eyepiece graticule to count and morphotype groupings of microorganisms as 1) coccoid forms, 2) rods, 3) filaments, 4) motile rods, 5) spirochetes or other cell types. Crevicular fluid flow measurements and clinical findings (pocket depth, and gingival inflammation) were also investigated. The following conclusions were drawn: The electrode with capillary was useful to determine the pH of the crevicular fluid clinically. A reduction in pH level was seen in deep pockets or severe gingival inflammation. A close correlation was seen between salivary and crevicular pH. The pH level was significantly positively related with the proportion of coccoid forms, but was negatively correlated with the proportion of motile organisms that are reported to be related with periodontal disease.

Dental Instruments↗
PubMed1989

[Clinical effects of chewing gum containing egg-white lysozyme and mace extract].

The purpose of this study was to evaluate the clinical effect of mace extract and egg-white lysozyme in two brands of chewing gum on gingival condition. Ever since mace extract containing dihydroguaiaretic acid was reported to inhibit the growth of Streptococcus mutans, plans were devised to include it in commercially available chewing gum. Before starting this study, two different types of experimental chewing gum containing mace extract or egg-white lysozyme were made up. A control was also prepared containing neither agent. The periodontal condition of 68 patients with gingivitis was determined based on PMA index (PMA), gingival index (GI), gingival bleeding index (GBI) and plaque scoring system (PSS) and randomly classified into three groups. Each group was instructed to use one or the other of the above type chewing gums after every meal. The results were as follows: 1. No clinical changes were observed in the control group during this study. 2. Gingival inflammation (PMA, GI, GBI) significantly improved as a result of using the experimental gums. 3. Plaque reduction was found only in the mece-extract gum group. 4. No clinical side effects were detected during this study.

Chewing Gum↗
PubMed1989

[Therapeutic embolization for bilateral renal angiomyolipomas with spontaneous rupture--a case report].

A 64-year-old woman of bilateral renal angiomyolipomas with spontaneous rupture, not associated with tuberous sclerosis was reported. Superselective embolization for tumors using ethanol was successfully performed, preserving normal renal function. Transcatheter embolization was considered the treatment of first choice for the urgent circumstances with spontaneous rupture.

Embolization, Therapeutic↗
PubMed1989

[A case of cutaneous T cell lymphoma improved with local administration of tumor necrosis factor].

A 63-aged woman with cutaneous T cell lymphoma successfully treated with local administration of tumor necrosis factor (TNF) was reported. She was admitted to our hospital because of tumors and subcutaneous nodules on her bilateral inner thigh. A pathological study of her skin of right inner thigh showed mononuclear atypical cells with hyperlobulated nuclei. In peripheral blood the same lymphoid cells were found. Immunohistochemical staining of these cells was positive for OKT-3 and OKT-4, but negative for OKT-8. No lymph node swelling and no visceral involvement were detected by a CT scan and a Echography of the chest and the abdomen. A diagnosis of cutaneous T cell lymphoma was made (stage IIb TNM classification). Although the chemotherapy of VEPA and CHOP was done, about 70% (PR) of the bilateral inner thigh tumors were retracted. Owing to the interstitial pneumonia aroused in the period of bone marrow suppression and cardiomyopathy after chemotherapy, we gave up further systemic chemotherapy. And then the local administration of TNF was done and the disappearance of the bilateral inner thigh tumors was obtained. Our therapy with local administration of TNF for CTCL in the first report.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1989

[Clinical study on anterior cruciate ligament reconstruction with the scaffold type artificial ligament (Leeds-Keio)].

Over 400 cruciate ligaments were reconstructed with the newly developed scaffold type Leeds-Keio artificial ligament made of polyester. In this survey, 146 ACL cases with over 36 months postoperative history were clinically and in some cases arthroscopically and histologically reviewed. In the first series of 80 cases, ACL was reconstructed intra-articularly alone, and in the second series of 66, extra-articular lateral stabilization was added to the intra-articular procedure. In the first series, the Lachman test was improved in 88%, anterior drawer sign in 80% and jerk test in 75%. In the second, these were improved to a much greater degree, so that the corresponding figures were 95%, 86% and in 91% respectively. There were no complications such as joint effusion, chronic synovitis and infection. Revision due to implant rupture was done in 8 cases. Postoperative arthroscopy showed coverage of the implant by new tissue and development into a biologically sound ligament by 12 months postoperatively. Histological examination of the biopsy specimen obtained in arthroscopy showed abundant collagenous fibres running longitudinally. These results indicate that the Leeds-Keio artificial ligament yields satisfactory clinical results.

Adolescent↗
PubMed1988

Placental anticoagulant proteins: isolation and comparative characterization four members of the lipocortin family.

Previously we isolated and characterized a placental anticoagulant protein (PAP or PAP-I), which is a Ca2+-dependent phospholipid binding protein [Funakoshi et al. (1987) Biochemistry 26, 5572] and a member of the lipocortin family [Funakoshi et al. (1987) Biochemistry 26, 8087]. In this study, three additional anticoagulant proteins (PAP-II, PAP-III, and PAP-IV) were simultaneously isolated from human placental homogenates prepared in the presence of 5 mM ethylenediaminetetraacetic acid. The isoelectric points of PAP-I, PAP-II, PAP-III, and PAP-IV were 4.8, 6.1, 5.9, and 8.1, respectively, and their apparent molecular weights were 32,000, 33,000, 34,000, and 34,500, respectively. Amino acid sequences of cyanogen bromide fragments of these proteins showed that PAP-III was a previously unrecognized member of the lipocortin family, while PAP-II was probably the human homologue of porcine protein II and PAP-IV was a derivative of lipocortin II truncated near the amino terminus. Comparative studies showed that all four proteins inhibited blood clotting and phospholipase A2 activity with potencies consistent with their measured relative affinities for anionic phospholipid vesicles. However, PAP-IV bound to phospholipid vesicles approximately 160-fold more weakly than PAP-I, while PAP-II and PAP-III bound only 2-fold and 3-fold more weakly. These results increase to six the number of lipocortin-like proteins known to exist in human placenta. The observed differences in phospholipid binding may indicate functional differences among the members of the lipocortin family despite their considerable structural similarities.

Amino Acid Sequence↗
PubMed1988

Mechanism-based isocoumarin inhibitors for trypsin and blood coagulation serine proteases: new anticoagulants.

Trypsin, porcine pancreatic kallikrein, and several blood coagulation enzymes, including bovine thrombin, bovine factor Xa, human factor Xa, human plasma factor XIa, human plasma factor XIIa, and human plasma kallikrein, were inactivated by a number of substituted isocoumarins containing basic functional groups (aminoalkoxy, guanidino, and isothiureidoalkoxy). 3-Alkoxy-4-chloro-7-guanidinoisocoumarins were found to be the most potent inhibitors for the coagulation enzymes tested with kobsd/[I] values in the range of 10(3)-10(5) M-1 s-1. 4-Chloro-3-isothiureidoalkoxyisocoumarins show high inhibitory potency toward porcine pancreatic kallikrein, human plasma kallikrein, human factor XIa, human factor XIIa, and trypsin with kobsd/[I] values of the order of 10(4)-10(5) M-1 s-1. The inhibition of these serine proteases by the substituted isocoumarins are time dependent, and the inactivation of trypsin by 3-alkoxy-4-chloro-7-guanidinoisocoumarins and 7-amino-4-chloro-3-(3-isothiureidopropoxy)isocoumarin occured concurrently with the loss of the isocoumarin absorbance. The complex formed from inactivation of trypsin by these two types of inhibitors was very stable and regained less than 4% activity in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (pH 7.5) after 1 day at 25 degrees C and regained 8-45% activity upon addition of buffered 0.29 M hydroxylamine. Trypsin inactivated by other inhibitors regained full activity upon standing or addition of hydroxylamine. Thrombin inactivated by 3-alkoxy-4-chloro-7-guanidinoisocoumarins was also quite stable and only regained 9-15% activity under similar conditions. These results are consistent with a proposed mechanism, where serine proteases inactivated by aminoalkoxyisocoumarins or isothiureidoalkoxyisocoumarins form acyl enzymes that will deacylate upon standing or addition of hydroxylamine. However, the acyl enzymes formed from 3-alkoxy-4-chloro-7-guanidinoisocoumarins or 7-amino-4-chloro-3-(3-isothiureidopropoxy)-isocoumarin will decompose further, probably through a quinone imine methide, to give an irreversibly inactivated enzyme by reaction with an active-site nucleophile such as His-57. The quinone imine methide intermediate may also react with a solvent nucleophile to give an acyl enzyme that can be reactivated by hydroxylamine. The inhibitors 4-chloro-7-guanidino-3-methoxyisocoumarin and 4-chloro-3-ethoxy-7-guanidinoisocoumarin have been tested as anticoagulants in human plasma and were effective at prolonging the prothrombin time. However, they are unstable in plasma (t1/2 = 4-8 min), and their in vivo utility may be limited.

Animals↗
PubMed1988

Highly sensitive peptide-4-methylcoumaryl-7-amide substrates for blood-clotting proteases and trypsin.

Seventy-four peptide amides of 7-amino-4-methylcoumarin (Mec) of the type Boc-Xaa-Yaa-Arg-NH-Mec were newly synthesized and tested to find specific substrates for blood-clotting proteases and trypsin. The Xaa and Yaa residues of these substrates have been replaced by 12 and 15 different amino acids, respectively. Among these peptides, the followings were found to be most sensitive substrates for individual enzymes: Boc-Asp(OBzl)-Pro-Arg-NH-Mec (kcat = 160 s-1, Km = 11 microM, kcat/Km = 15,000,000 M-1 s-1) for human alpha-thrombin, Z-less than Glu-Gly-Arg-NH-Mec (kcat = 19 s-1, Km = 59 microM, kcat/Km = 320,000 M-1 s-1) for bovine factor Xa, Boc-Gln-Gly-Arg-NH-Mec (kcat = 5.8 s-1, Km = 140 microM, kcat/Km = 42,000) for bovine factor XIIa, Boc-Asp(OBzl)-Ala-Arg-NH-Mec (kcat = 9.2 s-1, Km = 120 microM, kcat/Km = 77,000 M-1 s-1) for bovine activated protein C, and Boc-Gly-Phe-Arg-NH-Mec (kcat = 29 s-1, Km = 230 microM, kcat/Km = 130,000 M-1 s-1) for bovine plasma kallikrein. Moreover, Boc-Glu(OBzl)-Ala-Arg-NH-Mec (kcat = 46 s-1, Km = 370 microM, kcat/Km = 120,000 M-1 s-1) was newly found as a good substrate for human factor XIa. Bovine trypsin effectively hydrolyzed peptide-NH-Mec substrates containing Ala and Pro at the P2 site. The most reactive substrate was Boc-Gln-Ala-Arg-NH-Mec (kcat = 120 s-1, Km = 6.0 microM, kcat/Km = 20,000,000 M-1 s-1).

Amino Acids↗