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Biomedical subjects

T Kosugi

Publications and source records attributed to T Kosugi.

At least 55 records · Page 3Linked to original sources
PubMed1996

Effects of a synthetic N-terminal fragment of stanniocalcin on the metabolism of mammalian bone in vitro.

A synthetic peptide corresponding to the N-terminal amino acid residues of stanniocalcin (STC1-20) and including a region that is known to be an active site in teleosts was prepared and tested for its effects on the metabolism of mammalian bone in vitro. STC1-20 (10(-10)-10(-12) M) inhibited increases in the number of tartrate-resistant acid phosphatase-positive, multinucleated cells promoted by an N-terminal fragment of human parathyroid hormone (hPTH1-34) in cultures of murine hemopoietic cells. STC1-20 also slightly decreased the rate of loss of radioactivity from calvariae of fetal rats that had been prelabeled with 45Ca, both with and without stimulation by hPTH1-34. The accumulation of cAMP induced by hPTH1-34 in ROS 17/2.8-5 cells was suppressed by STC1-20 (10(-10)-10(-12) M). Treatment with STC1-20 (10(-11)-10(-13) M) caused increases of the rate of incorporation of [3H]proline into the collagenase-digestible protein of calvariae in newborn mice. From these results, it appears that STC1-20 has diverse effects on the metabolism of mammalian bone, causing a biphasic response. Such effects have not been observed with intact stanniocalcin or with materials from the corpuscles of Stannius and they are also different from the effects of hPTH1-34.

Acid Phosphatase↗
PubMed1996

Compartment analysis of cerebral glucose metabolism and in vitro glucose-metabolizing enzyme activities in the rat brain.

To clarify the relationship between cerebral glucose metabolic rate constants and glucose-metabolizing enzyme activities in the cerebral cortex, we evaluated the cerebral metabolic rate of glucose (CMRGlu), metabolic rate constants of [18F]-2-fluoro-2-deoxy-D-glucose (FDG) and related enzyme activities in the frontal cortex under normal and glucose metabolism-suppressed conditions. Applying a three-compartment four-parameter model, metabolic rate constants were obtained by dynamic positron emission tomography with FDG, and CMRGlu was calculated based on these rate constants. The glycolytic enzyme activities were determined by in vitro biochemical assay. Three days after ibotenic acid injection into the basal forebrain, CMRGlu was decreased in the ibotenic acid-treated frontal cortex as well as k3* (phosphorylation), while K1* (plasma to brain) showed no remarkable change. No significant reductions of the enzyme activities except for hexokinase activity were found in the frontal cortex. Regression analysis showed a significant positive correlation between k3* and the hexokinase activity. These results suggested that k3* in the compartment analysis reflects hexokinase activity.

Analysis of Variance↗
PubMed1996

In vivo trafficking of long-circulating liposomes in tumour-bearing mice determined by positron emission tomography.

Various kinds of long-circulating liposome, such as ganglioside GM1-, polyethyleneglycol- (PEG-), and glucuronide-modified liposomes, have been developed for passive targeting of liposomal drugs to tumours. To evaluate the in vivo behaviour of such long-circulating liposomes, we investigated the liposomal trafficking, especially early trafficking just after injection of liposomes, by a non-invasive method using positron emission tomography (PET). Liposomes composed of dipalmitoylphosphatidylcholine, cholesterol, and modifier, namely, GM1, distearoylphosphatidylethanolamine (DSPE)-PEG or palmityl-D-glucuronide (PGlcUA), were labelled with [2-18F]-2-fluoro-2-deoxy-D-glucose ([2-18F]FDG), and administered to mice bearing Meth A sarcoma after having been sized to 100 nm. A PET scan was started immediately after injection of liposomes and continued for 120 min. PET images and time-activity curves indicated that PEG liposomes and PGlcUA liposomes were efficiently accumulated in tumour tissues time dependently from immediately after injection. In contrast, GM1 liposomes accumulated less in the tumour as was also the case for control liposomes that contained dipalmitoylphosphatidylglycerol (DPPG) instead of a modifier. Long-circulating liposomes including GM1 liposomes, however, remained in the blood circulation and avoided liver trapping compared with control DPPG liposomes. These data suggest that PGlcUA and PEG liposomes start to accumulate in the tumour just after injection, whereas GM1 liposomes may accumulate in the tumour after a longer period of circulation.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1996

Habutobin releases plasminogen activator (U-PA) from bovine pulmonary artery endothelial cells.

Habutobin is a thrombin-like enzyme, contained in the venom of Trimeresurus flavoviridis, which has the strongest toxic effect in cases of habu bite. The present study was undertaken to examine the effect of habutobin on the release of plasminogen activators using cultured endothelial cells of the bovine pulmonary artery. The chemical characteristics of the plasminogen activators released into the conditioned medium were determined by fibrin autography and immunological analysis. A chromogenic substrate (S-2251) microassay was employed for quantitative estimation of the plasminogen activator activity in the conditioned medium and euglobulin fraction derived from the conditioned medium. The levels of plasminogen activator inhibitor released into the conditioned medium were determined by reverse fibrin autography. Fibrin autography revealed that cultured bovine pulmonary artery endothelial cells spontaneously released tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA) into the conditioned medium with no stimulus. Exposure of confluent cultures to 50 nM habutobin, however, induced a time-dependent increase in the level of plasminogen activator activity in both the conditioned medium and euglobulin fraction, and the plasminogen activator activity in the euglobulin fraction at 24 hr was significantly higher than that in the control (P < 0.05). Reverse fibrin autography demonstrated that the lysis-resistant zone of the supernatant of the euglobulin fraction (habutobin exposure) was wider than that in the case of no stimulus. These findings suggest that habutobin induced a time-dependent increase in the levels of plasminogen activator and plasminogen activator inhibitor concomitantly.

Animals↗
PubMed1996

Alpha 2-macroglobulin of rabbits inhibits the habutobin activity.

We have investigated whether alpha 2-macroglobulin (alpha 2M) of rabbits inhibits the activity of habutobin, a thrombin-like enzyme from Trimeresurus flavoviridis venom. Rabbit alpha 2M was purified with ultracentrifugation, gel filtration on Sepharose 6B and ion exchange chromatography on DEAE-Sephacel. Inhibitory effects of rabbit alpha 2M on habutobin was determined by the fibrin forming activity, digestion of A alpha chain of fibrinogen, and the release of fibrinopeptide A from fibrinogen. As a results, purified alpha 2M showed a single band with high molecular weight, around 800,000 mol. wt by means of polyacrylamide gel electrophoresis using PhastSystem. Besides inhibiting amidolytic and caseinolytic activity of porcine trypsin, it has inhibited the activity of habutobin: that is, in the presence of rabbit alpha 2M, fibrin forming activity of habutobin was decreased and habutobin-induced digestion of A alpha chain was inhibited. In addition, rabbit alpha 2M reduced habutobin-induced release of fibrinopeptide A from rabbit fibrinogen.

Animals↗
PubMed1996

Effect of Habu (Trimeresurus flavoviridis) antivenom on changes of hemostatic parameters following administration of crude venom from T. flavoviridis in rabbits.

We carried out experiments to examine the efficacy of Habu antivenom in relation to variations of hemostatic parameters induced by the administration of crude venom to rabbits. For neutralization of the crude venom by Habu antivenom, the correlation between the concentrations of Habu antivenom (Y) and crude venom (X) was expressed by the equation: Y = -0.473 + 0.539X. We examined the variations in hemostatic parameters in the state which crude venom was neutralized by Habu antivenom following the administration of crude venom (1 mg/kg) from Trimeresurus flavoviridis. Although the hemostatic parameters [level of fibrinogen, antithrombin III (AT-III) activity and alpha 2-plasmin inhibitor (alpha 2-PI) activity] underwent decreases after the administration of crude venom, they revealed fluctuations within the normal range after the Habu antivenom administration. The AT-III activity, however, decreased gradually until 90 min after the antivenom administration. These results suggested that Habu antivenom was effective in improving the abnormal coagulant activity induced by crude venom. However, the neutralization effect towards the coagulant activity of crude venom by the Habu antivenom did not continue for a long time and did not lead to recovery of the AT-III activity. Since excessive doses of antivenom can induce serious medical problems, we expect that simultaneous use of antivenom and AT-III preparation, instead of excessive and single use of Habu antivenom could provide a useful therapy for snake bites from the present study.

Animals↗
PubMed1996

Serial changes in titers of antibody to hepatitis B surface antigen after immunization of infants born to mothers with hepatitis B e antigen.

To evaluate the long-term protection provided by hepatitis B (HB) vaccine in a high-risk environment, we followed 50 infants born to mothers with hepatitis B e (HBe) antigen. These infants were immunized with a three-dose regimen of plasma-derived HB vaccine and were followed for up to 10 years. Two infants (4%) acquired hepatitis B virus (HBV) infection before 1 year of age. The 48 other infants remained hepatitis B surface (HBs) antigen-negative during the follow-up period. The geometric mean titers of serum antibody to HBs antigen rapidly decreased during the first 4 years but remained at protective levels throughout the follow-up period. These data suggest that protection against significant HBV infection lasts for at least 10 years. We conclude that the long-term protection afforded by plasma-derived HB vaccine is satisfactory and that a routine booster dose before 10 years of age is not necessary.

Female↗
PubMed1996

Production of a monoclonal dinitrophenyl-specific rat IgE and establishment of an IgE capture ELISA for estimating the concentration of rat IgE antibodies to dinitrophenyl-Ascaris suum.

A hybridoma producing monoclonal rat IgE antibodies of antidinitrophenyl (anti-DNP) specificity was generated by fusion of Sp2/0-Ag 14 (SP2) mouse myeloma cells and spleen cells from a DNP-Ascaris-sensitized Brown-Norway rat. Subsequently, the supernatant of the hybridoma (FE-3) was applied to an affinity column of DNP-bovine serum albumin-Sepharose 4B. The adsorbed protein fraction was pooled, concentrated, and further purified using Sephadex G-200. The molecular weight of the isolated protein was approximately 200,000 by SDS-PAGE, and the protein reacted with peroxidase (POD) mouse antirat myeloma IgE on Western blotting. Rabbit antibodies against DNP-specific rat IgE were also prepared by immunizing Japanese white rabbits with monoclonal DNP-specific rat IgE. These antibodies against DNP-specific rat IgE were applied to an affinity column of normal rat serum-Sepharose 4B and monoclonal DNP-specific rat IgG2b-Sepharose 4B to remove any other reactive substances apart from IgE contained in the serum proteins of the rat sensitized with DNP-Ascaris. On ELISA, it was found that the specificity of POD rabbit antibodies against DNP-specific rat IgE for monoclonal DNP-specific rat IgE was the same as that for rat myeloma IgE (IR 162). In addition, determinations of the monoclonal DNP-specific rat IgE revealed that the sensitivity of ELISA using POD-rabbit antibodies against DNP-specific rat IgE [POD-RA(DNP)RE] was higher than that using POD goat antibodies against rat myeloma IgE. Furthermore, an IgE capture ELISA employing the above-mentioned RA(DNP)RE was established for estimating the rat IgE antibodies to DNP-Ascaris suum. A good correlation was found between the antigen-specific IgE antibodies in the serum of Wistar rats estimated by this IgE capture ELISA and those estimated by passive cutaneous anaphylaxis.

Animals↗
PubMed1995

Pharmacokinetics of habutobin in rabbits.

Following the administration of habutobin, the fibrinogen level in the circulating blood of the rabbits decreased. These results showed that the activity of habutobin was retained in vivo. The plasma level of habutobin was determined by a ELISA-double sandwich method. The pharmacokinetics of habutobin from Trimeresurus flavoviridis venom was studied in rabbits following i.v. administration of 50 micrograms kg-1 of habutobin. The time course of the plasma concentration of habutobin fitted a two-compartment open model. The half-life of the distribution phase was 4.43 +/- 1.28 min and that of the elimination phase was 50.42 +/- 7.89 min. The area under the plasma concentration-time curve (AUC) was 38.69 +/- 6.68 micrograms min ml-1. The total body clearance was 3.82 +/- 1.08 ml min-1. When the steady state was reached, the concentration ratio of habutobin in the tissue (Ct) to that in the plasma (Cc), Ct:Cc was 0.47:1. These findings suggest that relatively little habutobin tended to remain in the tissue.

Animals↗
PubMed1995

Protective effect of thromboxane synthetase inhibitor on hypertensive renal damage in Dahl salt-sensitive rats.

1. The effects of OKY-046, a specific thromboxane (TX) synthetase inhibitor, on blood pressure, urinary excretion of TX and its release from blood platelets and renal papilla, and pathological change of glomeruli were evaluated in Dahl salt-sensitive rats. 2. Average daily intakes of OKY-046-treated rats were 0.93 mg/kg (low dose), 9.8 mg/kg (moderate dose), and 88 mg/kg (high dose). 3. Systolic blood pressure tended to decrease by 6.3, 11.4, and 10.9% in three OKY-treated groups. 4. OKY-046 suppressed the release of TX from platelets in a dose-dependent fashion. Both TX in urine and released from renal papilla decreased in OKY-treated groups with moderate and high dose. OKY-046 resulted no change in urinary excretion or release from renal papilla of prostaglandin E2 or 6-keto-prostaglandin F1alpha. 5. Glomerular sclerosis score decreased significantly in both groups treated with moderate and high doses of OKY-046. 6. An inhibition of renal TX synthesis by TX synthetase inhibitor has a protective effect on the development of hypertensive renal damage with minor antihypertensive effect in Dahl salt-sensitive rats.

Animals↗
PubMed1995

Plasminogen activators in tissue extract of aural cholesteatoma.

Using a biochemical technique, the authors characterized and identified the plasminogen activator (PA) derived from tissue extracts of six aural cholesteatomas. The results of fibrin zymography indicated that the tissue extracts of two cholesteatomas demonstrated two lytic zones on fibrin-agarose plates. One of the lytic zones was at about 72 kd, while the other zone was at about 64 kd. Using various goat immunoglobulin G (IgG)-containing antibodies (anti-human uterine tissue type PA (t-PA), anti-human low-molecular-weight (LMW) urokinase, and nonspecific goat IgG) and plasminogen-free fibrin-agarose plates, we confirmed that the cholesteatoma tissue extracts contained 72 kd t-PA and 64 kd urokinase type PA (u-PA). Furthermore, we measured the t-PA and u-PA activities in the tissue extracts selectively by parabolic rate assay. In order to estimate the PA activity, we developed optimal conditions for this assay. The specific t-PA activity ranged from 0.03 to 0.43 mIU/micrograms-protein and the specific u-PA activity ranged from 0 to 0.35 mIU/microgram-protein. The highest percentage of u-PA with respect to the total PA activity was 44.9%. However, in four of the six cases, we failed to detect u-PA activity. In the present study, we thus clarified the presence of PAs in tissue extracts of aural cholesteatomas. Furthermore, we confirmed that measureable u-PA occurred in some tissue extracts. We anticipate that the u-PA in inflammatory tissues plays an important role in the degradation of the extracellular matrix via the formation of plasmin and collagenases.

Cholesteatoma, Middle Ear↗
PubMed1995

[Role of renal thromboxane in salt-sensitive blood pressure rise in DOCA salt hypertension].

Urinary excretion and release of prostaglandins (PG) from isolated glomeruli and renal papilla of deoxycorticosterone acetate (DOCA)-treated rats fed with a normal salt (0.6% NaCl) diet and a high salt (4%NaCl) diet were determined. Mean blood pressure was significantly higher in the high salt diet group than in the normal salt diet group (146.2 +/- 2.3 vs 118.6 +/- 1.9 mmHg, p < 0.01). Urinary excretion of thromboxane (Tx) B2, stable metabolite of Tx A2, and 6-keto PG-F1 alpha, a stable metabolite of prostacyclin, increased significantly in the high salt diet group compared to the values of the normal salt diet group increased significantly by 104%, 55%, and 74% compared to those of the normal salt diet group, respectively. Release of 6-keto-PG F1 alpha from renal papilla of the high salt diet group decreased significantly, but there were no intergroup differences in the release of PG E2 and Tx B2. Stepwise multiple linear regression analysis showed that the significant contributory factors underlying the mean blood pressure were urinary excretion of Na (F = 14.187, p < 0.01) and release of Tx B2 from isolated glomeruli (F = 4.135, p < 0.05). These findings suggest that the manifestation of Tx synthesis in renal glomeruli has a predominant role in the salt sensitive pressor response of DOCA salt hypertension in rats.

Animals↗
PubMed1995

Effect of argatroban on the formation of artificial thrombus on dogs.

In the present study, we have investigated the effects of a synthetic antithrombin, Argatroban, and an antiplatelet agent. Ticlopidine hydrochloride, on the weight of artificial thrombus. These drugs at various concentrations were added to canine bloods, which were adjusted to 20%, 40% and 60% of haematocrit, and an artificial thrombus was formed using a modification of Chandler's method. Argatroban inhibited the formation of artificial thrombus, and this marked inhibition was observed especially in the experiment using blood with a high value of Ht. On the other hand, Ticlopidine hydrochloride did not inhibit the formation of artificial thrombus. From these results, it becomes clear that the mechanism of inhibitory action of Argatroban on artificial thrombus formation is based on the inhibition of thrombin activity and not on the inhibition of platelet aggregation. In addition, it is suggested that Argatroban inhibits the aggregation of red blood cells in the manner of direct or indirect action.

Animals↗
PubMed1994

Brain activation study by use of positron emission tomography in unanesthetized monkeys.

A system for the measurement of brain activity in conscious monkeys by positron emission tomography (PET) was established in the present study. The signal/noise ratio was maximal around 40 s for data acquisition in the PET scan with 15O-labeled water. When the monkey was stimulated by vibration and subtraction images of the data sets from regional cerebral blood flow (rCBF) changes in paired stimulation and control were superimposed on magnetic resonance images obtained from the same specimens, a somatotopic map corresponding to the sites stimulated was clearly demonstrated. Visual stimulation with a photic stimulator activated the corresponding regions of the primary visual cortex. Comparison of the activated sites and extents under the conscious state with those under anesthesia assured that the study is controllable; there was little unpredictable activation due to unlimited subject movement or to psychological effects.

Animals↗
PubMed1994

Application of a monoclonal antibody to estimate rabbit fibrinopeptide A released by habutobin.

We reported previously that habutobin, one of the type A thrombin-like enzymes, releases fibrinopeptide A alone from rabbit fibrinogen. To evaluate the effective action of habutobin in experiments using rabbit for the treatment of thrombosis, we attempted to develop an immunological method for measuring the fibrinopeptide A level in the circulating blood of rabbit. The purified rabbit fibrinopeptide A was coupled to keyhole limpet hemocyanin and BALB/c mice were immunized with the resultant fibrinopeptide A-hemocyanin conjugate. The spleen cells of an immunized mouse were fused with myeloma cells (P3-X63-Ag8-U1). As a result, one hybridoma (a-F-7) was selected, which secreted an antibody against rabbit fibrinopeptide A. Using this monoclonal antibody, we developed a competitive enzyme-linked immunoassay for estimating rabbit fibrinopeptide A. It was able to measure rabbit fibrinopeptide A contained in bentonite defibrinated plasma. This competitive enzyme-linked immunoassay should be useful for determining the fibrinopeptide A level in the circulating blood of rabbits, using plasma defibrinated by bentonite.

Adjuvants, Immunologic↗
PubMed1994

Enhancement of platelet aggregation in patients with nasopharyngeal carcinoma.

The ADP-induced platelet aggregation in 157 patients with nasopharyngeal carcinoma (NPC) in South China was estimated. The difference between the maximum platelet aggregation rate (MAR) in the control group and the MAR of patients with stage III/IV was significant. That is, the MAR at stage III and stage IV was significantly enhanced, as compared to that in the control group and that at stage I or stage II. The results of the present study indicate that ADP-induced platelet aggregation is enhanced in NPC patients of stage III and stage IV.

Adenosine Diphosphate↗
PubMed1994

Histamine release from rabbit platelets by platelet-activating factor (PAF).

In this study, we examined firstly whether PAF could activate washed platelets of rabbits and release ATP from the platelets, secondly whether activated platelets could release histamine, and thirdly whether certain antagonists of PAF could inhibit the release of histamine from platelets. The results obtained in these experiments may be summarized as follows. 1) Release of ATP was increased with enhanced platelet aggregation by PAF. At 3.4 x 10(-7) M of PAF, the maximal aggregation rate of platelets was about 70% and the concentration of released ATP was 1.6 x 10(-5) M. 2) After aggregated platelets induced by PAF had been sonicated and centrifuged, the resultant supernatant could cause contraction of the guinea pig ileum. This contraction was inhibited by the antihistaminic agents, cimetidine and pyrilamine. Furthermore, the histamine content of the supernatant was about 3.7 micrograms/ml (platelet count, 30 x 10(4)/microliter). 3) At 10(-5) M of CV-3988, the inhibitory ratio of PAF-induced aggregation was 35% and that of histamine release was 50%. On the other hand, at 5 x 10(-7) M of CV-6209, the inhibitory ratio of PAF-induced aggregation was 35% and that of histamine release was about 30%. From the above results, it was clear that PAF-activated platelets could release histamine. In addition, it is suggested that a direct relationship between PAF and platelets may exist in the process of allergic reactions, and histamine from PAF-activated platelets may modify allergic reactions.

Adenosine Triphosphate↗