Unique activation status of peripheral blood mononuclear cells at acute phase of Kawasaki disease.

Although Kawasaki disease (KD) is characterized by a marked activation of the immune system with elevations of serum proinflammatory cytokines and chemokines at acute phase, the major sources for these chemical mediators remain controversial. We analysed the activation status of peripheral blood mononuclear cells (PBMCs) by flow cytometry, DNA microarray and quantitative reverse transcription-polymerase chain reaction. The proportions of CD69+ cells in both natural killer cells and gammadeltaT cells at acute-phase KD were significantly higher than those at convalescent-phase KD. Microarray analysis revealed that five genes such as NAIP, IPAF, S100A9, FCGR1A and GCA up-regulated in acute-phase KD and the pathways involved in acute phase KD were related closely to the innate immune system. The relative expression levels of damage-associated molecular pattern molecule (DAMP) (S100A9 and S100A12) genes in PBMCs at acute-phase KD were significantly higher than those at convalescent-phase KD, while those of TNFA, IL1B and IL6 genes were not significantly different between KD patients and healthy controls. Intracellular production of tumour necrosis factor-alpha, interleukin-10 and interferon-gamma in PBMCs was not observed in KD patients. The present data have indicated that PBMCs showed a unique activation status with high expression of DAMP genes but low expression of proinflammatory cytokine genes, and that the innate immune system appears to play a role in the pathogenesis and pathophysiology of KD.

Two KaiA-binding domains of cyanobacterial circadian clock protein KaiC.

kaiABC, a gene cluster, encodes KaiA, KaiB and KaiC proteins that are essential to circadian rhythms in the unicellular cyanobacterium Synechococcus sp. strain PCC 7942. Kai proteins can interact with each other in all possible combinations. This study identified two KaiA-binding domains (C(KABD1) and C(KABD2)) in KaiC at corresponding regions of its duplicated structure. Clock mutations on the two domains and kaiA altered the strength of C(KABD)-KaiA interactions assayed by the yeast two-hybrid system. Thus, interaction between KaiA and KaiC through C(KABD1) and C(KABD2) is likely important for circadian timing in the cyanobacterium.

Structures and contribution to the antigenicity of oligosaccharides of Japanese cedar (Cryptomeria japonica) pollen allergen Cry j I: relationship between the structures and antigenic epitopes of plant N-linked complex-type glycans.

The oligosaccharide structures of Cry j I, a major allergenic glycoprotein of Cryptomeria japonica (Japanese cedar, sugi), were analysed by 400 MHz 1H-NMR and two-dimensional sugar mapping analyses. The four major fractions comprised a series of biantennary complex type N-linked oligosaccharides that share a fucose/xylose-containing core and glucosamine branches including a novel structure with a nongalactosylated fucosylglucosamine branch. Rabbit polyclonal anti-Cry j I IgG antibodies cross-reacted with three different plant glycoproteins having the same or shorter N-linked oligosaccharides as Cry j I. ELISA and ELISA inhibition studies with intact glycoproteins, glycopeptides and peptides indicated that both anti-Cry j I IgGs and anti-Sophora japonica bark lectin II (B-SJA-II) IgGs included oligosaccharide-specific antibodies with different specificities, and that the epitopic structures against anti-Cry j I IgGs include a branch containing alpha 1-6 linked fucose and a core containing fucose/xylose, while those against anti-B-SJA-II IgGs include nonreducing terminal mannose residues. The cross-reactivities of human allergic sera to miraculin and Clerodendron Trichotomum lectin (CTA) were low, and inhibition studies suggested that the oligosaccharides on Cry j I contribute little or only conformationally to the reactivity of specific IgE antibodies.

Antigenicity of the oligosaccharide moiety of the Japanese cedar (Cryptomeria japonica) pollen allergen, Cry jI.

Cry jI, a major allergenic glycoprotein of Cryptomeria japonica (Japanese cedar, Sugi), is the most common pollen allergen in Japan. Carbohydrate analysis and lectin staining indicated that Cry jI possesses the fucose/xylose-containing N-linked oligosaccharide which previously has been found in some plant glycoproteins. Rabbit polyclonal anti-Cry jI IgG antibodies were found to be highly cross-reactive with two other plant glycoproteins which have the same type of oligosaccharides, and the cross-reactivities were completely abolished on chemical deglycosylation of the glycoproteins. Enzyme-linked immunosorbent assay inhibition showed that the majority (up to 70%) of the anti-Cry jI rabbit IgG antibodies recognized the oligosaccharide moiety of Cry jI. The carbohydrate epitopes were found to be only partially involved in the binding of specific IgE antibodies from a pool of human patient sera.

[Cis-diamminedichloroplatinum(II) chemotherapy in advanced pancreatic carcinoma].

The effect of cis-diamminedichloro platinum (II) (CDDP) was evaluated in 14 patients with advanced pancreatic carcinoma. Prior chemotherapy was done in 7 cases and the remaining 7 were fresh cases. The drug was given at a dose of 80 mg/m2 I.V. every 3 weeks, with hydration and mannitol diuresis. Four cases out of 14 showed no change, while the remaining 10 cases showed progressive disease. The response rate was 0%. Most of the patients who showed myelosuppression had received prior chemotherapy. Non-hematologic toxicity occurred in 9 patients (64%) and consisted of nausea in 9, vomiting in 7 and anorexia in 9. Values of serum creatinine and BUN were transiently elevated.

[Serial changes in serum CA 19-9 levels in pancreatic cancer].

Levels of a new carbohydrate antigen CA 19-9, which is a monosialoganglioside identified by a monoclonal antibody, were measured in 41 patients with pancreatic cancer and these serial changes were investigated. To provide a contrast, the serum CEA levels were compared with the serum CA 19-9 levels. As the result of this study, it was found that the serial changes of the serum levels of CA 19-9 were more correlative to the clinical course of these patients with pancreatic cancer than those of the serum CEA levels.

A case of pancreatic cancer with liver metastasis which responded to chemotherapy.

A case of pancreatic cancer with liver metastasis is reported, in which chemotherapy had a marked effect, with the responses clearly documented. The patient was a 59-year-old male who experienced epigastric pain in February 1985, upper gastrointestinal X-ray examination revealing extragastric compression by the pancreas. He visited the National Cancer Center Hospital, Tokyo, in April 1985 and the diagnosis of pancreatic cancer with liver metastasis was made using the imaging procedures of ultrasonography, computed tomograph scanning and endoscopic retrograde cholangiopancreatography as well as by biochemical and serological tests. At the time a 5 cm tumor was palpable in the middle upper abdomen. The patient was treated with Cis-diaminedichloro platinum, tegafur, and 5-fluorouracil, successively, and the abdominal tumor gradually diminished, finally becoming impalpable. The response was evaluated as one of partial responses (PR) by ultrasonography, and the improvement substantiated by computed tomography and tumor markers.

Synthesis of plasmin substrates and relationship between their structure and plasmin activity.

The plasmin substrates, D-Ile-Phe-Lys-pNA (I), 3-MV-Phe-Lys-pNA (II), Ile-Phe-Lys-pNA (III), D-Pro-Phe-Lys-pNA (IV), CP-Phe-Lys-pNA (V) and Pro-Phe-Lys-pNA (VI), were synthesized by the conventional solution method and the kinetic parameters of their amidolysis by plasmin were determined. It was found that the free amino group of the D-amino acid in substrates (I) and (IV) made a contribution to an increment in affinity between the substrate and plasmin.

[Difference in clinical treatment between patients histologically diagnosed as group III by the old and new "group classifications" of gastric biopsy].

Since 1971, when the first draft of the "Group Classification", which classifies the atypism of histological structure in biopsy specimens of the stomach, was proposed by the Japanese Research Society For Gastric Cancer, this classification has come into wide use in Japan. It was, however, revised in 1983, and, according to the revised classification, group III was defined as general histological findings in which it is difficult to make a differentiation between benign and malignant by biopsy specimens. Consequently, the new group III includes, various borderline histologies, in addition to the old group III, which had been defined as the histological features corresponding to those observed in gastric adenoma in biopsy specimens. At the National Cancer Center Hospital, 13,909 gastric biopsies were performed during the period between 1973 and 1982. By retrospective review of these, histological findings in 247 lesions of 231 cases corresponded to group III by the old classification (adenomatous type) and in addition to these, 54 lesions of 54 cases to group III by the new classification (non-adenomatous type). We compared the endoscopic and pathological features between the two types, and the following results were obtained: The false-negative rate of malignancy in the non- adenomatous type (24%) was much higher than that in the adenomatous type (6%). The difference between the two may suggest that, with the adoption of the new group classification, clinical treatment of the patients with group III becomes more complicated due to the increase of the false-negative rate. Endoscopically, most (84%) of the lesions in the adenomatous type were seen as polypoid, while in the non-adenomatous type, depressed lesions were dominant (80%). And, endoscopic details of the polypoid or depressed appearances were mostly different between the two types. These nuances of endoscopic appearance between adenomatous and non-adenomatous types are applicable to decisions regarding, adequate clinical treatment for patients diagnosed as group III by the new "group classification." Good communication between the endoscopist and pathologist is indispensable.

Diagnosis of early gastric cancer seen as less malignant endoscopically.

In order to make the endoscopic diagnosis of early gastric cancer more accurate, we determined the endoscopic and clinicopathological characteristics of the 129 cases of early gastric cancer with less malignant appearance among the 978 cases of the disease with a solitary lesion detected at the National Cancer Center Hospital during the period between 1962 and 1981. According to findings required to make a differential diagnosis, the endoscopic appearance of the 978 cases were divided into the following four groups: 170 cases of polypoid, 587 of ulcerative, 119 of gastritis-like and 102 of advanced cancer-like. Among the four groups, the frequency of cases with less malignant appearance was quite high in the gastritis-like group. The lesion in this group was defined as the cancerous lesion which shows only superficial mucosal abnormality, not associated with any polypoid or ulcerative components. Chronologically, it has been rapidly increasing in recent years. These two paradoxical phenomena in the gastritis-like group, that is, "high frequency of less malignant appearance" and "increase in number of cases" not only indicate the inadequacy of the conventional criteria for the recent cases but also show that the gastritis-like malignancy is detectable. New additional endoscopic criteria for the gastritis-like malignancy, for example, irregular margin of the shallow mucosal depression, disproportion of the mucosal granularity, irregularity on the erythematous change and discoloration are essential in order to detect much more cases with less malignant appearance.

[An autopsy report of pancreatic carcinoma following the achievement of significant prolongation of survival, despite the lack of a definitive diagnosis].

A 56-year-old female presented with chief complaints of epigastric discomfort and jaundice. Various examinations led to a differential diagnosis of carcinoma of she pancreas vs. malignant lymphoma. Chemotherapy and radiotherapy achieved a 50% reduction of the tumor mass and the disappearance of her jaundice. The patient survived for twenty-seven months following initial presentation. On post-mortem examination, she was found to have had carcinoma of the head of the pancreas. We studied the relationship between chemotherapy and the survival time in histologically diagnosed, unresectable pancreatic cancer at this hospital over the last ten years.

Selective inhibition of thrombin by (2R,4R)-4-methyl-1-[N2-[(3-methyl-1,2,3,4-tetrahydro-8-quinolinyl++ +) sulfonyl]-l-arginyl)]-2-piperidinecarboxylic acid.

The potency of thrombin inhibition by 4-methyl-1-[N2-[(3-methyl-1,2,3,4-tetrahydro-8-quinolinyl)-sulfony l]- L-arginyl]-2-piperidinecarboxylic acid (MQPA) depended on the stereoconformation of the 2-piperidinecarboxylic acid moiety. Ki values for bovine alpha-thrombin were 0.019 microM with (2R,4R)-MQPA, 0.24 microM with (2R,4S)-MQPA, 1.9 microM with (2S,4R)-MQPA, and 280 microM with (2S,4S)-MQPA. (2R,4R)-MQPA of the four stereoisomers of MQPA was also the most potent inhibitor for other trypsin-like serine proteases with Ki values of 5.0 microM for trypsin, 210 microM for factor Xa, 800 microM for plasmin, and 1500 microM for plasma kallikrein. Examination of the potency of thrombin inhibition by arginine derivatives related to MQPA in structure suggested the presence of a specific binding site for the carboxamide portion (C-terminal side). The relative inhibitory potency of the four stereoisomers of MQPA for trypsin was nearly identical with that for thrombin, suggesting that the specific binding site for the carboxamide portion is present in both enzymes. Modification of thrombin by phosphopyridoxylation or the presence of heparin did not significantly alter the binding of MQPA.

Thrombin inhibitors. 3. Carboxyl-containing amide derivatives of N alpha-substituted L-arginine.

A series of N alpha-(arylsulfonyl)-L-arginine amide derivatives having carboxamide N-substituents with a carboxyl group was prepared and tested as inhibitors of the clotting activity of thrombin. The most inhibitory compounds were obtained when a carboxyl group was introduced into the carbon next to the amide nitrogen of N alpha-(arylsulfonyl)-L-arginine amide derivatives, e.g., N alpha-(arylsulfonyl)-L-arginyl-N-butyl-, N-(methoxyethyl)- or N-(tetrahydrofurfuryl)glycine and 4-alkyl-1-[N alpha-(arylsulfonyl)-L-arginyl]-2-piperidinecarboxylic acid, with an I50 of 1-3 X 10(-7) M.

Thrombin inhibitors. 1. Ester derivatives of N alpha-(arylsulfonyl)-L-arginine.

A series of N alpha-(arylsulfonyl)-L-arginine esters was prepared and tested as inhibitors of the clotting activity of thrombin. N alpha-Dansyl-L-arginine methyl ester was the most inhibitory of the N alpha-(arylsulfonyl)-L-arginine methyl esters. The most potent inhibitors were the n-propyl and n-butyl esters of N alpha-dansyl-L-arginine with an I50 of 2 X 10(-6) M. Esters of unsaturated straight-chain alcohols with a chain length of four carbons were also as inhibitory as the n-butyl ester. The inhibitors were hydrolyzed by thrombin and trypsin more slowly than N alpha-tosyl-L-arginine methyl ester.

Thrombin inhibitors. 2. Amide derivatives of N alpha-substituted L-arginine.

A series of N alpha-(arylsulfonyl)-L-arginine amide derivatives with substituted or unsubstituted naphthalene and heterocyclic compounds as the N alpha-substituent was prepared and tested as inhibitors of the clotting activity of thrombin. N-n-Butyl and N-n-butyl-N-methyl derivatives of N alpha-dansyl-L-arginine amide were the most inhibitory of N-alkyl and N,N-dialkyl derivatives of N alpha-dansyl-L-arginine amide. Their inhibitory effect was as potent as that of N alpha-dansyl-L-arginine-n-butyl ester with an I50 of 2 X 10(-6) M. N alpha-Substituted naphtalenesulfonyl-L-arginine amide derivatives of 4-methyl- and 4-ethylpiperidine also showed a potent inhibition with an I50 of 10(-7) to 10(-6) M. The most potent inhibitior in this study was 1-[N alpha-(4,6-dimethoxynaphthalene-2-sulfonyl)-arginyl]-4-methylpiperidine, with an I50 of 7.5 X 10(-8) M. Arginine amide derivatives of 4-methyl- or 4-ethylpiperidine with tetralin or an oxygen-containing heterocyclic compound as a N alpha-substituent showed an inhibition with an I50 less than 10(-5) M. N-Monosubstituted derivatives of N alpha-dansyl-L-arginine amide were not hydrolyzed at all by thrombin and were hydrolyzed very slowly by trypsin, and N,N-disubstituted derivatives were not hydrolyzed at all by both enzymes.