Purification, characterization and molecular cloning of Cha o 1, a major allergen of Chamaecyparis obtusa (Japanese cypress) pollen.

Pollen of Chamaecyparis obtusa (Japanese cypress) is one of the causes of allergic pollinosis in Japan. A major allergen of the pollen designated Cha o 1, was purified by two-step ion exchange chromatography. Cha o 1 was separated into four components with molecular masses of 48.5 kDa and 52.0 kDa, each with pIs of 6.77 and 6.82. The 23-residue N-terminal sequence of Cha o 1 was determined and shown to have high identity with that of Cry j 1, a major allergen of Cryptomeria japonica pollen. cDNA coding for Cha o 1 was cloned by hybridization screening using Cry j 1 cDNA as a probe. One of the cDNA clones, pCHA-1 was sequenced and found to code for a putative 21-residue signal peptide and a 354-residue native protein with a derived molecular mass of 38.1 kDa. The deduced amino acid sequence of Cha o 1 showed 79-80% identity with those of Cry j 1. These findings were consistent with observations of a close crossreaction between the two allergens. Homology analyses revealed that Cha o 1 had 46-49% identity with Amb a 1 families and Amb a 2, the major allergens of short ragweed. Cry j 1 has pectate lyase enzyme activity, suggesting that Cha o 1 may have the same enzyme activity as Cry j 1.

Preoperative radioactive therapy for esophageal carcinoma. Randomized evaluation trial in eight institutions.

A cooperative, prospective, randomized study to evaluate the effectiveness of preoperative irradiation in curatively resected esophageal carcinoma was performed in 364 cases in eight institutions from August 1982 to November, 1983. Based on the survival curves, postoperative irradiation alone was superior to preoperative plus postoperative irradiation. Because of the progression of the disease and complications after operation, there were many inevaluable cases in this study. However, analysis revealed there was no bias caused by a greater number of unfavorable patients being selectively included in inevaluable cases in the postoperative irradiation only group. Our data question the value of preoperative irradiation being performed in addition to resection plus postoperative irradiation.

Relation between the hormonal and epidemiological aspects of esophageal cancer in Japan.

Esophageal cancer with endemic distribution of high-risk areas in the world is also known to be more prevalent in the male--and in the aged-populations than in their counterparts. The hormonal characteristics of esophageal cancer patients and the geographical distribution of high and low-risk areas in Japan were comparatively investigated in search of the etiology of this neoplasia. A case-control study of 14 urinary steroids indicated that cancer patients of both sexes were associated with an acceleration of cortisone to hydrocortisone conversion within their bodies. Evidence was presented to suggest that the above state of glucocorticoid excess could be related to the production of hyperplasia as well as malignant transformation in the epithelial cells of the esophagus. The 10 highest risk areas and the 7 lowest risk areas were clearly separated by the 2 fault lines of the Japan Islands, a finding to suggest that some microelements of the soil play a role in conditioning the cancer risk. The possible linkage of the observed hormonal characteristics of Japanese patients to both the geochemistry of the soil and the genesis of this neoplasia is discussed in the light of the competitive relationship between the intake of vitamin C and endogenous nitrosation.

Interaction of fibrinolytic activity between the tracheobronchial secretion and circulating blood of rats.

In order to clarify the origin and release mechanism of plasminogen activator in the tracheobronchial secretion of rats, electrophoretic analysis of the secretion and studies on the effects of certain vasoactive drugs on the activator activity in the secretion were carried out. From the results of electrophoretic analysis of the tracheobronchial secretion in non-treated rats, protease inhibitor composed of glycoprotein was not contained in the secretion in contrast to the circulating blood, but a protein of low molecular weight like albumin in the circulating blood was contained in the secretion. Furthermore, after injection of noradrenalin, the blood pressure was temporarily elevated and the fibrinolytic activity of the euglobulin fraction in the circulatory blood was also increased. Subsequent to this elevation of fibrinolytic activity in the circulating blood, the fibrinolytic activity in the tracheobronchial secretion increased. Based on these results, it is suggested that the increase of fibrinolytic activity in the circulating blood was due to increased release of plasminogen activator from the vascular wall, and the increased fibrinolytic activity of the tracheobronchial secretion was caused by a consequent increased transudation of plasminogen activator from the circulating blood into the tracheobronchial lumen.

Properties of fucoidan from Cladosiphon okamuranus tokida in gastric mucosal protection.

To elucidate the anti-ulcer potential of Cladosiphon fucoidan, anti-peptic activity, bFGF stabilizing activity and inflammatory properties of this and related substances were investigated. Anti-peptic activity was observed with this and other sulfated polysaccharides such as dextran sulfate, carrageenan, and Fucus fucoidan. However, non-sulfated polysaccharides such as mannan and dextran did not exert the anti-peptic activity. The loss of bFGF bioactivity was prevented by all sulfated polysaccharides tested except chondroitin sulfate, at pH 7.4 and at pH 4.0. At pH 2.0, only heparin protected the bFGF activity. The generation of superoxide by macrophages and PMNs was stimulated by dextran sulfate, carrageenan, and Fucus fucoidan, whereas Cladosiphon fucoidan, heparin and chondroitin did not. Dextran sulfate, carrageenan, and Fucus fucoidan also stimulated the secretion of TNFalpha from macrophages, while Cladosiphon fucoidan did not. Thus, Cladosiphon fucoidan is a sulfated polysaccharide without inflammatory action. These results suggest that Cladosiphon fucoidan is a safe substance with potential for gastric protection.

Application of cryo-sapphire knife in rapid frozen ultrathin sectioning for electron microscopy.

Conventional sectioning of frozen and fixed tissues is usually performed using the glass knife. Although the glass knife is economical, it has several disadvantages. In this study, the Authors have examined the sectioning quality of the sapphire knife in frozen tissues as an alternative to the glass knife. It was concluded that the sapphire knife has better quality and durability than the conventional glass knife in cutting frozen tissues.

The role of fibrinolytic enzymes in inflammation and paranasal mucous membrane.

The distribution and role of tissue plasminogen activator (TA) and proactivator (PA) in various diseases of the nasal and paranasal cavity were investigated. The stronger the inflammatory and proliferative response of the paranasal mucous membrane, the weaker was the fibrinolytic activity of TA. The fibrinolytic activity of PA tended to be stronger than TA activity. It is considered that PA may play an important role in inflammatory enlargement and proliferation of the paranasal mucous membrane, but does not play an important role in carcinogenic enlargement and proliferation of the nasal and paranasal mucous membrane.

Studies on proactivator from the paranasal mucous membrane in chronic sinusitis.

We succeeded in differentiating the proactivator (PA) in tissue extract of paranasal mucous membrane with chronic sinusitis by the gel filtration technique. Furthermore, the results demonstrated that PA in the tissue extracts of the paranasal mucous membrane with chronic sinusitis and antrochoanal polyp was unrelated to the antigenicity of plasminogen. In particular, it was clarified that the tissue extract of antrochoanal polyp as a source of PA was not related to the antigenicity of plasminogen.

Differences in physicochemical properties between tissue plasminogen activators from human paranasal mucous membrane and pig heart.

In order to clarify the physicochemical property of tissue plasminogen activator (TA), tissue extracts of human paranasal mucous membrane and pig heart were studied by the biochemical techniques. The studies by gel filtration revealed that two plasminogen activators of different molecular weight were present in the extract of human paranasal mucous membrane. The existence of tissue plasminogen activator with a low molecular weight (LMW-TA) has not previously been reported. This molecular weight of this compound was lower than that of cytochrome c. On the other hand, the molecular weight of tissue plasminogen activator from pig heart (PH-TA) was similar to that of ovalbumin, about 46,000 daltons, as estimated by Sephadex G-150 gel filtration. From the physicochemical property of LMW-TA, it is suggested that the LMW-TA from paranasal mucous membrane with chronic sinusitis was produced by proteases, i.e. trypsin-like enzyme, present in the mucous membrane with chronic inflammation.

Control of Arthus tonsillitis by the administration of an antiplasminic agent.

Previous reports have shown that fibrinolytic activity was increased in the circulatory blood of rabbits with Arthus tonsillitis. In this study, t-AMCHA was administered intravenously to rabbits, in order to clarify its role as an antiplasminic agent in the control of Arthus tonsillitis. First, to establish the proper method of administration of t-AMCHA and the quantity to be administered, a preliminary estimation was made of the requisite concentration of t-AMCHA in the circulating blood and tonsillar tissue. Then macroscopic observations and histopathological studies were performed on the tonsils of rabbits with Arthus tonsillitis after t-AMCHA administration at various doses. Finally, after injection of the antiplasminic agent, estimations were made of the plasma fibrinolytic activity in the blood of the rabbits under study. It was decided that administration of t-AMCHA at 3-hourly intervals could maintain a constant concentration of the substance in the tonsillar tissue. The marked hyperaemia and bleeding which were observed macroscopically and histopathologically at 1-2 days after the onset of tonsillitis, disappeared from the parenchyma of the tonsils and pharyngeal tissue surrounding the tonsils in the group receiving regular injections of t-AMCHA at 3-hourly intervals. After estimating certain parameters of the fibrinolytic system in the blood, it was found that fibrinolytic activity decreased and whole plasmin was not consumed as a result of the administration of t-AMCHA during the early stage of tonsillitis. The process of Arthus tonsillitis can thus be controlled by the administration of the antiplasminic agent.

Antiplasmin activity in carrageenin-induced inflammation of rats.

The significance of inhibitors in the coagulation-fibrinolysis system was examined by inducing experimental inflammation with carrageenin. Fast-acting plasmin inhibitor, alpha 2-plasmin inhibitor (alpha 2-Pl), was recently discovered in human blood and it was stated that such alpha 2-Pl represents the most sensitive and specific inhibitor of plasmin. The possible role of alpha 2-Pl has been studied in detail in vitro and in vivo in relation to the thrombolytic condition and bleeding based on hyperfibrinolysis. However, the significance of alpha 2-Pl has not been clarified for the inflammatory state and diseases. In the present study in order to clarify the possible role of alpha 2-Pl in inflammatory responses, the antiplasmin activity in the plasma and local fluid (exudate) was investigated using an animal model of inflammation. After subcutaneous injection of carrageenin into the dorsum of rats, on day 7 and 21, the antiplasmin activity in the plasma was significantly higher than that in the plasma of non-treated rats (p less than 0.001). However, the antiplasmin activity in the plasma on day 7 was not significantly different from that on day 21. On the other hand, the antiplasmin activity in the exudate on day 21 was significantly lower than that on day 7 (p less than 0.001). Based on ;these results, it is suggested that the antiplasmin in the inflammatory exudate was consumed during the inflammation to form the hard wall of the carrageenin pouch.

The detection of fibrinogen (or FgDP)-protamine sulphate complex in vitro and in the circulating blood.

In order to clarify the pathophysiological significance of the fibrinogen (FgDP)-protamine complex, the interaction between fibrinogen and protamine sulphate was studied in vitro and in vivo. Using the electrophoretic technique, an attempt was made to determine whether the complex could be detected in vitro and in the circulating blood. From the present studies, it has been clarified in in vitro experiments that the slower mobility of two peaks on immunoelectrophoresis represented a complex of protamine sulphate and fibrinogen (FgDP). Furthermore, the peak of slower mobility on crossed immunoelectrophoresis appeared in the circulating blood of a rabbit receiving administration of excess protamine sulphate. That is, in vivo experiments showed that a complex of protamine sulphate and fibrinogen (FgDP) could be detected in the circulating blood.

Differentiation of proactivator from tissue plasminogen activator with low molecular weight in the paranasal mucous membrane.

It has been shown that large amounts of fibrinolytic enzyme are contained in tissue extracts of the paranasal mucous membrane in patients with chronic sinusitis. However, the fibrinolytic enzyme in the tissue extracts has not yet been characterized by biochemical techniques. In the present study, proactivator was differentiated from tissue plasminogen activator with low molecular weight in extracts of the paranasal mucous membrane and the existence of the proactivator was thus demonstrated in tissue extracts. By analogy with the proactivator in antrochoanal polyps, this proactivator may play an important role in the proliferation and enlargement of the paranasal mucous membrane.

Effect of oligofucose derivatives on acetic acid-induced gastric ulcer in rats.

This study attempted to enhance the anti-ulcer activity of fucoidan from Cladosiphon okamuranus TOKIDA by chemical modification with a hydrophobic group. The suitable number of fucose residues in the effective compound was also clarified to obtain a compound of constant quality. Degraded fucoidans were coupled with several hydrophobic groups via Schiff bases, and their anti-ulcer activities were determined by acetic acid-induced ulcer models in rats. Size-fractionated oligofucose was also modified and assayed for anti-ulcer activity. Among the modified oligofucoses, only the oligofucose-dodecylaniline combination (OFDA) significantly promoted ulcer healing. The effective dose was 0.2 mg/kg/d. The most suitable number of fucose residues in the compound for the anti-ulcer activity was determined to be around 12. We succeeded in enhancing the anti-ulcer activity of Cladosiphon fucoidan by modification with dodecylaniline. The activity of this compound was comparable or greater than that of typical anti-ulcer agents. By determination of the optimal OF chain length for the anti-ulcer activity of OFDA, it became possible to obtain OFDA of constant quality.

Morphological study of parafollicular cells and the parathyroid gland in the house shrew (Suncus murinus).

A morphological study of parafollicular cells in the thyroid gland and parathyroid gland of the house shrew (Suncus murinus) was made. The results indicated that (1) there were two pairs of parathyroid glands which were located in the upper part of the house shrew thyroid gland, (2) the volumes of the house shrew parathyroid glands ranged from 0.014 to 0.079 mm3, (3) the number of parafollicular cells along the follicles was largest in the upper part of the thyroid lobe, while no parafollicular cells were present in the isthmus, and (4) there were about 20 parafollicular cells per 100 follicular cells and 1.519 parafollicular cells per follicle. The number of parafollicular cells per 100 follicular cells was thus about 5 times larger than that in rats and the number of parafollicular cells per follicle about 2.5 times larger than that in rats.

[Phase II study on peplomycin for esophageal cancer].

For the purpose of verifying the effectiveness of peplomycin, one of the derivatives of bleomycin, against carcinoma of the esophagus and the safety of it, the analysis of the data for total 113 cases collected from 25 institutions in Japan was made. The results are as follows. It was effective in 19 out of 74 evaluable cases of carcinoma of the esophagus (25.7%). In case of treatment with peplomycin alone, it was effective in 6 out of 39 cases (15.4%). In case of the combination treatment with peplomycin and some other therapy, it was effective in 13 out of 35 cases (37.1%). As for the side effects, the incidence of fever was the highest in both the cases of peplomycin alone and the combination treatment such as 39.6% and 37.0%, respectively. Anorexia, nausea, vomiting, respiratory symptoms and tiredness were found in relatively many cases. In the clinical laboratory tests, the vital capacity after the treatment tended to be lower than that before the treatment, but there was little change in the hematological tests, pulmonary function test and renal function test.

[Predictive evaluation and efficient management of medical examinations using Mahalanobis Taguchi System Method].

UNLABELLED: The Mahalanobis-Taguchi System (MTS), an evaluative technique used in quality engineering, was utilized in a system analysis of medical examinations to determine practicability, to differentiate cost reduction benefits of specific components for increasing efficiency, and to determine possibility of utility on forecasting of future health condition of individuals. METHODS: 1. Medical examination date over a two year period were utilized to differentiate subject into two groups: Group A--Healthy or normal; and Group C--Patient group requiring treatment or currently undergoing treatment. The Mahalanobis distance for Group C from the Mahalanobis space of Group A was calculated for each subject to determine differentiability. 2. Utilizing the MTS method specific components of the examination system were selected and analyzed for effectiveness in diagnosis and screening to enhance efficiency of the examination. 3. Group A subjects (determined from the two year data), were analyzed using the data at the end of 1 year to develop the Mahalanobis space and the Mahalanobis distance at the 2nd year point was calculated and analyzed. RESULTS: 1. Subjects who were classified as group A at both 1st and 2nd year points numbered 159. With a cutoff point of 2.0 for Mahalanobis distance 98.1% of the healthy subjects (Group A) were classified as belonging to the healthy group, and 91.9% of C group as belonging to the patient group. 2. Blood pressure, GOT, GPT, gamma-GTP, T Chol, TG, UA, Cr, BS, HbA1c, Hb, and MCH were found to be effective components in the examination. Based on this result the items included in the examination can be reduced by approximately 60%. 3. With cutoff value of 1.5, the results of 90% of subjects 1 year later could be predicted. The prediction that 15% of Group C subjects would improve to join Group A could be made. Overall, the results 1 year later could be predicted at a level of 90% accuracy. CONCLUSION: These results suggest that the Mahalanobis-Taguchi System technique can accurately assess medical examination quality and can also be used, with reasonable accuracy, to predict the condition of individuals one year later. Therefore, the MTS can be used to help determine which patients should be receiving the examination 1 year later. In addition, components of the examination can be assessed using the MTS for benefits and costs to determine levels of essentiality of components. As a result, it can be used an effective tool for development and improvements in the examination system. With further development in the technique, it may become possible for use as a tool in disease diagnosis and treatment.

[Study of washout rate in patients with chronic renal failure by dipyridamole 201Tl myocardial scintigraphy--comparison with hypertrophic cardiomyopathy].

Fourteen patients with chronic renal failure (CRF) were studied by dipyridamole 201Tl myocardial scintigraphy. In this study, patients with coronary heart disease were excluded by clinical findings. 201Tl SPECT was performed at 8 min after injection of dipyridamole (0.56 mg/kg) and at a 3-hr delayed interval, and washout rates (WR) were calculated. Because UCG showed left ventricular hypertrophy (thickness; 13 +/ 2 mm) in CRF, the WR values were compared with those of hypertrophic cardiomyopathy (HCM) (n = 9). In CRF, an average WR was 54.2 +/- 6.8% and was significantly higher than that in normal (46.2 +/- 9.1%, p < 0.05, n = 9), though WR in HCM was significantly low (26.9 +/- 11.4%, p < 0.01). Hematocrit value was 24.5 +/- 3.2% in CRF and was significantly lower than that in normal (37.8 +/- 3.6%, p < 0.001). On the other hands, hemoconcentration was observed in HCM (hematocrit; 44.6 +/- 3.6%, p < 0.001). WR in CRF, HCM and normal had significant correlation with hematocrit values (r = -0.796, p < 0.001). These results suggest that anemia or hemoconcentration may affect coronary blood flow and WR, and this should be considered when diagnosing coronary heart disease in CRF or HCM.

[Study of surgical treatment of advanced small lung cancer].

We analyzed 15 patients with small but advanced primary lung cancer operated from 1965 to 1988. The size of tumors were less than 2.0 cm in largest dimension and their pathological stages III A, III B and IV. The histological types were adenocarcinoma in 13 cases and squamous cell carcinoma in 2 cases. Nine patients (69%) had elevated levels of preoperative serum CEA. The prognosis of patients with intrapulmonary metastases was better than that of those with pleural dissemination. In adenocarcinoma patients with mediastinal lymph node metastases, the survival rate of cases with small cancer less than 2.0 cm in diameter was higher than those with large cancer more than 5.0 cm in diameter.

Poly (lactic/glycolic acid) microspheres containing antigen as a novel and potential agent of immunotherapy for allergic disorders.

In order to establish a safer and simpler antigen administration method in immunotherapy, we prepared biodegradable microspheres containing antigen and evaluated its safety and efficacy using guinea pigs. Poly (lactic/glycolic acid); (LGA) microspheres containing ovalbumin (OA) were fabricated by solvent evaporation. Over 70% of the OA was released from the microspheres within 3 days, and release was completed within 14 days in vivo. The local tissue reactions to the OA-LGA microspheres were apparently weaker than those to OA-alum. Repeated injections of high dose OA-LGA microspheres to OA-sensitized guinea pigs (high-LGA group) for 8 weeks at intervals of 2 weeks elicited an excellent therapeutic effect, i.e. a significant increase in the threshold value of antigen inhalation test, with a significant increase in IgG2 blocking antibody. The therapeutic efficacy of the high-LGA group was comparable to the conventional immunotherapy model (conventional group) and was superior to the antigen-alum model (alum group). We concluded that administration of antigen-LGA microspheres could become a new immunotherapeutic method for allergic disorders, being safer and requiring a lower frequency of antigen injections than the conventional method.