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Biomedical subjects

T Kosugi

Publications and source records attributed to T Kosugi.

At least 73 records · Page 4Linked to original sources
PubMed1993

Fibrin binding, fibrinolytic and fibrinogenolytic activity of plasminogen activator derived from the paranasal mucous membrane of humans.

It is known that large amounts of plasminogen activator (PA) are contained in tissue extracts of the human paranasal mucous membrane (PMM) with chronic sinusitis. The present study was undertaken to isolate and purify the PA in tissue extracts of PMM. Furthermore, the purified PA was identified as to whether it was of the tissue type or urokinase (UK) type, and some of its fibrinolytic characteristics were determined in comparison with those of urokinase. As starting material, extracts of acetone powder of PMM with chronic sinusitis were used, and Zn-imminodiacetate affinity chromatography, and ultrafiltration were carried out to separate and purify the PA from the PMM. The PA was purified to a 107-fold increase in specific activity. The molecular weight of the PA was estimated to be 65,000 to 70,000 d by gel filtration using Sephacryl S-200. The purified PA was stable in the range of pH 8.0 to 9.0. Using S-2288, a synthetic substrate, the Michaelis constant (Km) of the purified PA was estimated to be 0.11 mmol. The binding of the purified PA to fibrin was stronger than that of UK, while the fibrinogenolytic activity of the purified PA was not stronger than that of UK. Based on these results, the purified PA was identified as a tissue-type plasminogen activator (t-PA). From the kinetic data, it was identified as being of the two-chain variety. It is considered that, as a thrombolytic agent, t-PA derived from the PMM could be more useful than UK.

Animals↗
PubMed1993

Inhalation of platelet-activating factor increases respiratory resistance in rats: determination by means of an astograph under nonanesthetized conditions.

Intratracheal administration of platelet-activating factor (PAF) to dogs, baboons, and humans has been shown to induce hyperreactivity of the airways and contraction of the smooth muscle. However, it has not yet been reported whether intratracheal administration of PAF to rats induces hyperreactivity. In the present study, the authors estimated the respiratory resistance of rats during intratracheal administration of PAF in order to evaluate the reactivity of the airways to PAF. In both the nonsensitized group and the sensitized group of rats, intratracheal administration of PAF induced an increase in respiratory resistance. The results obtained clarify that responsiveness to PAF exists in the airways of rats.

Administration, Inhalation↗
PubMed1993

A modification of the ELISA-double sandwich method for estimating the concentration of habutobin.

A monoclonal antibody to the thrombin-like enzyme, habutobin, was produced. An ELISA-double sandwich method employing this monoclonal antibody was devised as a method for determining the habutobin concentration in vitro. However, the absorbance of the control sample in such an ELISA-double sandwich procedure was too high to estimate low levels of the enzyme. The present study therefore attempted to establish a reliable ELISA-double sandwich method in which the absorbance of the control sample was lower than previously, and which had a high sensitivity, in order to determine the habutobin concentration in vitro and in vivo. The modification of the ELISA-double sandwich technique employing the monoclonal antibody against habutobin, the purified rat IgG against habutobin and POD-mouse anti-rat IgG2a, provided a reliable means of determining the habutobin levels in the circulating blood of rabbits.

Animals↗
PubMed1992

Production of a monoclonal antibody against the thrombin-like enzyme, habutobin, from Trimeresurus flavoviridis venom.

We succeeded in producing a monoclonal antibody to the thrombin-like enzyme, habutobin, which was purified from crude venom of the snake Trimeresurus flavoviridis. The monoclonal antibody obtained belonged to IgG1, and its light chain consisted of a kappa-chain. The monoclonal antibody reacted specifically with habutobin and crude venom from T. flavoviridis but did not react with human thrombin or bovine thrombin on Western blotting. The concentration of habutobin and crude venom of T. flavoviridis, in vitro, could be measured by means of ELISA using the monoclonal antibody. Furthermore, the ELISA-double sandwich method employing this monoclonal antibody may represent a reliable method for determining the habutobin levels in the circulating blood.

Animals↗
PubMed1992

Evaluation of provocation tests in tonsillogenic focal infection from the standpoint of fibrinolytic activity.

In this paper, we carried out provocation tests by means of ultra-short wave stimulation in patients with focal infection including palmoplantar pustulosis and examined the changes in fibrinolytic activity of the circulating blood. In the positive group with palmoplantar pustulosis, the t-PA level was slightly increased after the provocation and the FDP level in the positive group was increased. On the other hand, the AT-III activity in the positive group with palmoplantar pustulosis was decreased and the antiplasmin activity in the positive group was increased. The changes of antiplasmin activity observed in this experiment did not correspond with the pathophysiological explanation of an increased fibrinolytic activity in the circulating blood. It was suggested that a positive provocation test is accompanied by an increase in fibrinolytic activity in the circulating blood of patients with focal infection of the tonsil, and the increase in fibrinolytic activity is closely related to the positiveness of the provocation test.

Antibodies, Monoclonal↗
PubMed1992

Presence of urokinase-type plasminogen activator (u-PA) in tissue extracts of antrochoanal polyp.

Using a biochemical technique, the authors characterized and identified a plasminogen activator (PA) derived from tissue extracts of antrochoanal polyp (AP) and paranasal mucous membrane (PMM) with chronic sinusitis. The results of fibrin zymography indicated that the tissue extracts of AP revealed two lytic zones and that those of PMM revealed a single lytic zone on fibrin-agarose plates. One of the AP zones exhibited the same relative mobility as the PMM zone (molecular weight: 65 kd), while the other AP zone had a smaller molecular weight (about 54 kd). Goat immunoglobulin G (IgG) fraction of antihuman uterine tissue-type plasminogen activator (t-PA) inhibited the 65-kd lytic zones of AP and PMM. Antihuman low-molecular-weight urokinase inhibited only the 54-kd lytic zone of AP, and nonspecific goat IgG failed to inhibit any of the lytic zones. On the other hand, 10(-2) mol trans 4-(aminomethyl)cyclohexane-carboxylic acid (t-AMCHA) inhibited all of the lytic zones. No lytic zones could be observed on plasminogen-free fibrin-agarose plates. These findings confirmed that the tissue extracts of PMM contained t-PA, and that those of AP contained both t-PA and urokinase-type plasminogen activator (u-PA). In addition, it appeared that u-PA in inflammatory tissue was related to proliferative changes of the mucous membrane.

Humans↗
PubMed1992

[Evaluation of the sensitized condition of patients with allergic diseases in Okinawa using the MAST allergy system].

We determined, using the MAST system, specific IgE antibodies to allergens in the circulating blood of 127 patients with bronchial asthma, allergic rhinitis and atopic dermatitis in Okinawa. The positive rates to inhalant allergens in all patients examined by the MAST system were as follows: Dermatophagoides farinae 65%, house dust 58%, cat epithelium 17% and Japanese cedar 9%. In addition, the positive rates to food allergens found in all patients were as follows: wheat 16%, shrimp 14%, egg white, rice and crab 12%. The average number of positive allergens in the patients with atopic dermatitis was larger than that in those with allergic respiratory diseases only. The above five specific IgE antibodies were detected simultaneously in 12 (9%) of the 127 patients. In addition, the average number of overlapping positive allergens was 2.0 in all patients. From these results, it was suggested that the number of overlapping positive allergens in patients on Okinawa is smaller than in other areas of Japan as compared with other papers.

Adolescent↗
PubMed1992

Changes of rabbit platelet function following simultaneous administration of Ticlopidine hydrochloride and Argipidine (MD-805).

Ticlopidine hydrochloride and Argipidine were administered simultaneously to rabbits and the changes in platelet function and coagulation-fibrinolysis were determined. Ticlopidine hydrochloride and Argipidine did not give rise to an additive or synergistic effect on the ADP-induced platelet aggregation. However, simultaneous administration of Argipidine and Ticlopidine hydrochloride significantly inhibited the collagen-induced platelet aggregation, as compared to the effect of single administration of Ticlopidine hydrochloride at 60 min after intravenous administration of Argipidine. Furthermore, at 90 min after the intravenous administration, the PAF-induced platelet aggregation in the simultaneous administration was significantly different from that in each individual administration. These results suggested that the effect of simultaneous administration on the platelet aggregation was dependent largely on the effect of Ticlopidine hydrochloride alone.

Animals↗
PubMed1991

Plasminogen activator derived from tonsillar lymphocytes of rabbits with arthus tonsillitis.

As an experimental model of human tonsillitis, Arthus tonsillitis of the rabbit was produced. In a previous paper, we demonstrated that an increased fibrinolytic activity in the circulating blood and increased proteolytic activity of the local tissue with Arthus tonsillitis were observed. In the present study, the mechanism of the increased proteolytic activity at the local site (tonsil with Arthus-type tonsillitis) was examined using cell cultures of lymphocytes originating from the tonsil with Arthus-type inflammation. That is, lymphocytes isolated from tonsil tissue of the rabbit were cultured in a culture medium for the short term. After culture, the used medium was collected and the presence of plasminogen activator in the culture medium and lymphocytes was investigated. It was clarified that plasminogen activator did exist in the culture medium of lymphocytes originating from the tonsil of rabbits with Arthus tonsillitis. Furthermore, addition of T cell growth factor to the culture medium of lymphocytes derived from non-treated tonsil induced the development of plasminogen activator in that culture medium. However, addition of B cell growth factor did not lead to the development of plasminogen activator activity in the culture medium. These results suggest that plasminogen activator originating from lymphocytes of the tonsil may play an important role in the increased proteolytic activity observed at the local site of Arthus tonsillitis.

Animals↗
PubMed1991

Renal cyst formation as a complication of primary distal renal tubular acidosis.

Morphologic studies using renal sonography and CT scanning were performed in 6 patients with primary distal renal tubular acidosis who were followed for 6 to 18 years. Nephrocalcinosis and renal cysts were identified in all patients. Multiple bilateral renal cysts were detected in 3 patients, and unilateral single or several renal cysts were detected in 3 patients. Size and number of the renal cysts in 1 patient did not change over 5 years of alkali therapy. Although the precise mechanism for renal cyst formation has not yet been established, renal cysts may be a common complication of primary distal renal tubular acidosis.

Acidosis, Renal Tubular↗
PubMed1991

Changes in parameters of the coagulation-fibrinolysis system and platelet function after OKY-046 administration to patients with ruptured aneurysm of the cerebral artery.

The influence of the TXA2-synthetase inhibitor OKY-046 (Xanbon) on haematological findings for the coagulation-fibrinolysis system and platelet aggregation was investigated in patients with subarachnoid bleeding during or after the administration. Changes in alpha 2-PI activity and the levels of fibrinogen, t-PA and PAI antigen were observed. Especially, PAI activity and PAI antigen were found to be significantly increased as compared with levels before the administration. On the other hand, the platelet aggregation induced by various agents and the activity of AT-III were not greatly altered after the administration of OKY-046.

Adenosine Diphosphate↗
PubMed1991

Changes of rabbit platelet function on simultaneous administration of ticlopidine hydrochloride and OKY-046.

Ticlopidine hydrochloride 180 mg was given orally to rabbits and OKY-046 50 mg was simultaneously administered intravenously. Ticlopidine inhibited the platelet aggregation induced by ADP when the Ticlopidine was orally administered separately, but the platelet aggregation induced by PAF, collagen and arachidonic acid (A.a.) was not significantly decreased. Simultaneous administration of Ticlopidine and OKY-046, as compared to administration of Ticlopidine alone, led to a significant decrease in the platelet aggregation induced by A.a. and collagen. The simultaneous administration did not give rise to an additive or synergistic effect on the platelet aggregation induced by ADP.

Adenosine Diphosphate↗
PubMed1990

Identification of a plasminogen activator derived from nasopharyngeal carcinoma.

The activity of plasminogen activator (PA) in tissue extracts from nasopharyngeal carcinoma (NPC) was determined by means of the fibrin plate method. Development of PA activity was observed in 16 out of 25 cases investigated. Furthermore, using tissue extract of NPC with PA activity, characterization and identification of the activator were carried out by means of electrophoretic analysis, fibrin zymography and immunological analysis. The molecular weight of this PA was found to be 38,000 daltons. Additionally, a urokinase type of plasminogen activator was contained in the tissue extracts.

Adult↗
PubMed1990

Induction of airway hyperresponsiveness in allergic rats.

Brown-Norway rats (male) were sensitized with both dinitrophenylated-bovine serum albumin (DNP-BSA) and Bordetella pertussis simultaneously in order to induce airway hyperresponsiveness (AHR) as the first sensitization. At five days, DNP-BSA was inhaled as a booster into the airways under thiopental anaesthesia. At eight days, inhalation of antigen markedly increased the tracheal pressure (TP) in sensitized rats (11.9 +/- 1.6 cmH2O) and slightly increased TP in non-sensitized rats (1.1 +/- 0.4), the difference between the two groups being significant (p less than 0.001). Twenty-four hours after antigen challenge, the airway responsiveness to ACh in sensitized rats was markedly increased to about 4-fold as compared to that in non-sensitized rats. Inhalation of dinitrophenylated-ovalbumin failed to increase the airway responsiveness to ACh in rats sensitized with DNP-BSA, although a marked increase in TP was induced immediately after antigen challenge. We thus succeeded in preparing a model of AHR by employing a new procedure of sensitization.

Acetylcholine↗
PubMed1989

Effect of ultrasonic nebulization of Miraclid on the proteolytic activity in tracheobronchial secretions of rats.

Experimental studies of acute inflammation of the tracheobronchial lumen of rats suggest that protease inhibitor increases in tracheobronchial secretions in order to control inflammation. Recent studies have shown that the polyvalent protease inhibitor, Miraclid, derived from human urine, is useful for treating DIC and acute pancreatitis. In view of this information, local administration of Miraclid was expected to diminish acute inflammation of the respiratory tract by creating a favorable balance in the protease-antiprotease system. Before the chemotherapeutic use of locally administered Miraclid, the inhibitory activity of Miraclid on various proteases was first estimated in vitro. Administration of Miraclid by means of ultrasonic nebulization was then investigated in rats. The results can be summarized as follows. 1. During ultrasonic nebulization, the inhibitory activity of Miraclid on protease was decreased by means of mechanical stimulation in comparison to the activity before nebulization. 2. Compared to administration of physiological saline into the tracheobronchial lumen, administration of Miraclid by means of ultrasonic nebulization decreased the fibrinolytic activity in tracheobronchial secretions.

Aerosols↗
PubMed1987

Some chemical properties of tissue plasminogen activator purified from paranasal mucous membrane.

Plasminogen activator (PA) was purified from an acetone powder preparation of paranasal mucous membrane with chronic sinusitis, and some chemical properties of the purified PA were investigated in this paper. Zn-imminodiacetate affinity chromatography, lysine sepharose affinity chromatography and ultrafiltration for concentrating a PA fraction were consecutively performed to purify the PA from the acetone powder preparation. Finally, gel filtration was performed using Sephacryl S-200 in order to estimate the molecular weight of the purified PA. The purified PA in this experiment showed a stronger affinity to fibrin than urokinase did. The molecular weight of the purified PA was estimated to be 65,000 to 70,000 daltons as determined by Sephacryl S-200 gel filtration. The Km of the purified PA was 0.11 mM. From these results, it is apparent that the PA purified from an acetone powder preparation of paranasal mucous membrane belongs to the class of tissue type plasminogen activators (t-PA).

Chronic Disease↗
PubMed1987

Fibrinolytic activity in medium from tissue culture of paranasal mucous membrane.

It is known that a remarkable fibrinolytic activity of plasminogen activator (PA) can be seen in extracts of wet tissue and acetone powder preparation of paranasal mucous membrane evidencing chronic sinusitis. However, the origin of the PA in extract of paranasal mucous membrane has not yet been clarified up to the present time. In this experiment, using a tissue culture of paranasal mucous membrane, it was observed that two species of cells, epithelial cells and fibrocytes, proliferated in the implanted tissue. PA was isolated from the conditioned medium on the fifth day after culture. From these results, it appears that the PA may be released from epithelial cells and/or fibroblasts.

Culture Techniques↗