Discovery of cockroach specimens of the genus Squamoptera (Ectobiidae: Pseudophyllodromiinae) from Okinawa, Japan, showing wing polymorphism.

Cockroach specimens of the genus, Squamoptera were collected from the Iriomote island of Okinawa prefecture, Japan. The morphological features of the specimens were characterized as having a white band on the dorsal surface of its thorax, its tegmen reduced into a tiny scale-like structure and the hindwing was absent. Ocelli was also absent and the small compound eyes not extending to apex of the head nor to the frontal face but extend further lower than the base of the antennae. When the specimens were reared in the laboratory, besides the short wing form, the long wing form began to appear in the rearing colony. In our reproductive biological study, we observed that hatching of the ootheca from the short wing female takes about 30 days, with an average of 6.6 nymphs being hatched from one ootheca. The male to female ratio of the offspring was 36:30. However, the frequency appearance of the offspring from the ootheca of the short wing female was 98.5% short wing and 1.5% long wing form. Our specimens occasionally show body polymorphism in the form of individuals having long wings instead of the usual short one. The long wing form does not show the white band on the dorsal surface of its thorax.

Discovery of cockroach specimens of the genus Squamoptera (Ectobiidae: Pseudophyllodromiinae) from Okinawa, Japan, showing wing polymorphism

@# Cockroach specimens of the genus, Squamoptera were collected from the Iriomote island of Okinawa prefecture, Japan. The morphological features of the specimens were characterized as having a white band on the dorsal surface of its thorax, its tegmen reduced into a tiny scale-like structure and the hindwing was absent. Ocelli was also absent and the small compound eyes not extending to apex of the head nor to the frontal face but extend further lower than the base of the antennae. When the specimens were reared in the laboratory, besides the short wing form, the long wing form began to appear in the rearing colony. In our reproductive biological study, we observed that hatching of the ootheca from the short wing female takes about 30 days, with an average of 6.6 nymphs being hatched from one ootheca. The male to female ratio of the offspring was 36:30. However, the frequency appearance of the offspring from the ootheca of the short wing female was 98.5% short wing and 1.5% long wing form. Our specimens occasionally show body polymorphism in the form of individuals having long wings instead of the usual short one. The long wing form does not show the white band on the dorsal surface of its thorax.

Bactericidal activity and oral pathogen inactivation by electromagnetic wave irradiation.

AIMS: The aim of this work was to clarify the effects of electromagnetic wave irradiation (EMWI) on oral bacterial pathogens. METHODS AND RESULTS: A Gram-negative (Porphyromonas gingivalis) or Gram-positive (Streptococcus mutans, S. intermedius, Enterococcus faecalis) bacterial suspension was irradiated by EMW apparatus (500-1000 kHz, 5-15 times, 1 s time(-1) ). Quantification of survival bacteria by CFU counting revealed that EMWI exhibited marked bactericidal activity against all tested bacteria and bactericidal activity at 500 kHz increased in an irradiation number-dependent manner. After EMWI at 500 kHz, scanning electron microscopic observations showed that the chain of S. mutans cells was shortened after 5 irradiations and the outlines of bacterial cells (S. mutans and P. gingivalis) were unclear after 5-10 irradiations. EMWI inhibited the inductive effect of S. mutans on pro-inflammatory cytokine production in human monocytes and this inhibitory effect was comparable with that of heat-killed bacteria. Furthermore, using an enzyme activity assay, EMWI partially inactivated the activities of gingipains from P. gingivalis. CONCLUSIONS: These findings demonstrated that EMWI has inactivation and bactericidal activities against single microbial species among four kinds of oral pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Electromagnetic wave irradiation may be applicable for medical disinfection and sterilization, such as refractory periapical periodontitis.

Cloning of cDNA encoding a soybean allergen, Gly m Bd 28K.

A cDNA clone encoding a soybean allergen, Gly m Bd 28K, has been isolated. The clone has a 1567-bp cDNA insert with a 1419-bp open reading frame and a 148-bp 3'-untranslated region, followed by a polyadenylation tail. The open reading frame was shown to encode a polypeptide composed of 473 amino acids. The chemically determined amino acid sequences of the peptides obtained from the allergen, including its N-terminal peptide, were shown to be contained in the N-terminal region of the amino acid sequence deduced from the cDNA, showing that the first half of the cDNA encodes the allergen with a preceding segment of 21 amino acids. The peptide fragment including the allergen was expressed as a fusion protein with glutathione S-transferase in Escherichia coli and immunoblotted with the sera of soybean-sensitive patients and the monoclonal antibody against the allergen. Furthermore, homology analyses demonstrate that the polypeptide for the cDNA exhibits high homology with the MP27/MP32 proteins in pumpkin seeds and the carrot globulin-like protein. This finding suggests that the polypeptide may consist of a 21-amino acid segment as a part of the signal peptide and the proprotein, which may be converted to two mature proteins, Gly m Bd 28K and a 23-kDa protein, during the development of soybean cotyledons.

Occurrence of IgE antibody-recognizing N-linked glycan moiety of a soybean allergen, Gly m Bd 28K.

BACKGROUND: It has been reported that N-linked glycan moieties of glycoproteins function as IgE-reactive determinants. Gly m Bd 28K, a soybean allergen, was a glycoprotein with glycan moieties, which are supposed to be the Man(3)GlcNAc(2) backbone with the beta1-->2 xylose and alpha1-->3 fucose branches. The purpose of the present study was to examine the IgE-binding ability of the glycan moiety of Gly m Bd 28K in the binding reaction with patients' sera. METHODS: A peptide containing the glycan moiety was prepared from Gly m Bd 28K by digestion with lysyl endopeptidase. The binding site of the glycan moiety was determined by amino acid sequence analyses. The glycan moiety of the allergen was characterized using anti-horseradish peroxidase antibody (anti-HRP) recognizing the N-linked glycan moieties of glycoproteins. The binding of patients' IgE antibodies with their glycan moiety was examined by an immunostaining technique using the glycopeptide and its deglycosylated peptide derived from Gly m Bd 28K. RESULTS: The binding site of the glycan moiety in Gly m Bd 28K was shown to be its Asn20 residue. Gly m Bd 28K did react with anti-HRP and the sera of soybean-sensitive patients, but the binding of IgE antibodies was inhibited by the preincubation with anti-HRP. Moreover, the glycopeptide also reacted with the sera of soybean-sensitive patients, but its deglycosylated peptide did not react with any IgE antibodies of patients' sera. CONCLUSIONS: The specific IgE antibodies recognizing the N-linked glycan moieties of Gly m Bd 28K and other glycoproteins with homologous glycan moieties occur in the sera of soybean-sensitive patients. It was indicated that the N-linked glycan moieties such as that of Gly m Bd 28K may be one of the common IgE-reactive determinants distributed in various plant food proteins.

Immunochemical characterization of ovomucoid from Japanese quail egg white using monoclonal antibodies.

The ovomucoid of Japanese quail egg white is known as a proteinase inhibitor or protein component responsible for egg allergy. In order to characterize the antigenic properties of ovomucoid in relation to its molecular structure, we have prepared two monoclonal antibodies, E9 (IgG1) and E11 (IgG1), recognizing distinct epitopes from each other. These monoclonal antibodies bound to the SDS/mercaptoethanol-treated ovomucoid, but not to the reductively carboxymethylated and pyridylethylated ovomucoid. By immunoblotting analysis of the peptic digests of ovomucoid, it was shown that E9 bound to the fragment consisting of two domains, I and II, of the ovomucoid, and that E11 reacted with the fragment containing domain III. These results indicate that the antigenic activity depends on the conformational structure of domains tightly folded by disulfide linkages. Ovomucoids from hen and duck were also recognized by both the antibodies, although having less affinity compared to the one from Japanese quail. These antibodies proved to be effective in establishing an enzyme-linked immunosorbent assay system for quantitative analysis of quail ovomucoid.

Quantitative analysis of Gly m Bd 28K in soybean products by a sandwich enzyme-linked immunosorbent assay.

A sandwich enzyme-linked immunosorbent assay for the soybean allergen, which consists of a monoclonal antibody (D4) as the fixing (first) antibody and another peroxidase-conjugated monoclonal antibody (C5) as the second, has been developed. Both D4 and C5 monoclonal antibodies strongly bound to the guanidine/HCl-denatured allergen, Gly m Bd 28K. Therefore the samples used in the present experiment were extracted with sodium phosphate buffer (pH 8.0) containing 6 M guanidine and 10 mM 2-mercaptoethanol, then completely dialyzed against phosphate-buffered saline (PBS). The dialyzed samples were subjected to the assay. Various soybean products were observed to contain the allergen at high concentrations, such as soybean protein isolate (SPI), tofu, kori-dofu, and yuba, but its content in soy milk and abura-age were found to be low. In fermented products such as natto, soy sauce, and miso, and even in the processed foods with soybean protein isolate (SPI), the allergen was not detected. These results were also confirmed by an immunoblotting technique with D4.

[Unilateral multiple tumorous lesions of the parotid gland].

Multifocal tumors within the same parotid gland are very rare. We treated 13 patients with multiple tumorous lesions within the unilateral parotid gland. We evaluated the multiple nodules by CT-sialography or magnetic resonance imaging (MRI). These imagings showed clearly two or more distinct nodular-appearing lesions. Recurrent pleomorphic adenoma (6 patients) was predominant, followed by Whartin's tumor (3 patients). The other lesions were two differential parenchymal tumors (polymorphous low grade adenoma/adenoma) within the same gland, a malignant lymphoma, a squamous cell carcinoma metastatic to the gland, and a tuberculous lesion. On palpitation, 9 of the patients had an unilateral tumor, one a palpable parotid mass in the gland, and the other four had two or more tumors in the unilateral gland. The patients with intra-parotid lymph node and metastatic lesions had extra-parotid cervical adenopathy. The clinical features and the differential diagnosis of the unilateral multiple tumorous lesions of the parotid gland are discussed.

Purification of characterization of soybean allergen Gly m Bd 28K.

At least 15 allergenic proteins have been found in soybean using the sera of soybean-sensitive patients with atopic dermatitis [T. Ogawa et al., J. Nutr. Sci. Vitaminol., 35, 555-565 (1991)]. In the present study, a monoclonal antibody (mAb) against Gly m Bd 28K, one of the major allergens of soybean, was prepared, and Gly m Bd 28K was purified from defatted soybean flakes by five purification steps, including immunoaffinity chromatography with the mAb as a ligand. The purified allergen was found to be a glycoprotein with a molecular mass of 26 kDa. During the purification process the allergen was converted to more acidic proteins with the same molecular mass, suggesting that the allergen is unstable. The sugar composition and amino acid sequence of Gly m Bd 28K suggest that the allergen is a new glycoprotein with an Asn-linked sugar moiety. The distribution of the allergen in soybean products was examined by an immunoblotting technique with the mAb.

Micro-assay method for evaluating the allergenicity of the major soybean allergen, Gly m Bd 30K, with mouse antiserum and RBL-2H3 cells.

A micro-assay method for evaluating the allergenicity of soybean allergen was developed by using the mouse antiserum against Gly m Bd 30K, a major soybean allergen, and RBL-2H3, a rat mucosal mast cell line. The antiserum against Gly m Bd 30K was prepared by subcutaneously immunizing BALB/c mice with the allergen. The behavior by affinity-chromatography and the properties against heat treatment show that the reaginic activity of the antiserum resided in the IgE antibody specific for Gly m Bd 30K. The developed assay method is shown to be useful for simulating IgE mediated type-I allergy and to be highly sensitive for detecting the allergen.

Reduction of the allergenicity of soybean by treatment with proteases.

Soybeans were treated with proteases to reduce allergenicity. By immunoblotting with a monoclonal antibody against a major soybean allergen (Gly m Bd 30K), two of eight proteases so far tested were selected to achieve a reduction in allergenicity. Both antigenicity to the monoclonal antibody and allergenicity to the sera from soybean-sensitive patients proved to be markedly reduced by processing with either protease. Thus, soybeans treated with an appropriate protease may possibly be supplied as a hypoallergenic foodstuff for patients.

Effects of grinding and tableting on physicochemical stability of an anticancer drug, TAT-59.

The effects of grinding and tableting on the physicochemical stability of TAT-59, (E)-4-[1-[4-[2-(dimethylamino)ethoxy]phenyl]-2-(4-isopropyl) phenyl]-1-butenyl]phenyl monophosphate, were studied. The crystallinity of TAT-59 ground in a planetary ball mill for 0-120 min or compressed at 0-4500 kg/cm2 was evaluated by X-ray diffraction analysis and differential scanning calorimetry (DSC). The surface of TAT-59 was measured under a scanning electron microscope (SEM). The physicochemical stability of TAT-59, ground or compressed, was determined by measurements of water content, crystallinity and the amount of hydrolysis product, DP-TAT-59, formed. The crystallinity of ground TAT-59 decreased with increasing grinding time, and the amount of DP-TAT-59 increased with decrease in the crystallinity. Similar to ground TAT-59, the crystallinity of TAT-59 tablet gradually decreased with increasing compression pressure, and the amount of DP-TAT-59 tended to increase with decreasing crystallinity. These findings suggested that the decrease of the crystallinity of TAT-59 by mechanical force, such as grinding and tableting, raised the drug's reactivity and affected its stability.

Epitope analysis of soybean major allergen Gly m Bd 30K recognized by the mouse monoclonal antibody using overlapping peptides.

The epitopes of the major soybean allergen, Gly m Bd 30K, recognized by mouse monoclonal antibodies H6 and F5 were investigated by using synthetic peptides bound to pins. The epitopes are shown to be localized in peptide 31QGGCGRGWAFSATGAIEA48 for H6, and in 115DKVTIDGYETLIMSDEST132 for F5.

Identification of the glycosylation site of a major soybean allergen, Gly m Bd 30K.

A major soybean allergen, Gly m Bd 30K, is a glycoprotein. A peptide containing a sugar chain was isolated from an alpha-chymotrypic hydrolyzate of the allergen. The amino acid sequence analysis of the peptide showed that its sugar chain binds to the Asn170 residue. Furthermore, the sugar moiety of the allergen and peptide was shown to consist of mannose, N-acetylglucosamine, fucose, and xylose at a molar ratio of 3 : 2 : 1 : 1. These results indicate that Gly m Bd 30 K is a N-linked glycoprotein.

Alpha-subunit of beta-conglycinin, an allergenic protein recognized by IgE antibodies of soybean-sensitive patients with atopic dermatitis.

One of the major allergenic proteins in the soybean 7S-globulin fraction, which was recognized by sera of about 25% soybean-sensitive patients with atopic dermatitis, was identified as alpha-subunit of beta-conglycinin. The IgE antibodies recognizing the alpha-subunit did not show a cross-reactivity against both alpha'- and beta-subunits known to be highly homologous to alpha-subunit, and also against other allergenic components identified in soybeans [Ogawa et al., J. Nutr. Sci. Vitaminol., 35, 555-565 (1993)]. The IgE-binding site(s) was estimated to be located on the peptide 232-283 of alpha-subunit.

Measurement of Gly m Bd 30K, a major soybean allergen, in soybean products by a sandwich enzyme-linked immunosorbent assay.

By a sandwich enzyme-linked immunosorbent assay, a soybean major allergen, Gly m Bd 30K, in soybean products was measured. The allergen occurred at high concentrations in soy milk, tofu, kori-dofu, and yuba, but its content in kinako was small. No allergen was found in fermented foods such as miso, shoyu, and natto. The allergen was clearly shown to occur in meat balls, beef croquettes, and fried chicken that contained soybean protein isolate.

Infantile spasms with predominantly unilateral cerebral abnormalities.

To investigate the pathogenesis of infantile spasms, 12 children underwent a combination of neurophysiologic and neuroimaging studies including brainstem evoked potentials and single photon emission computed tomography with 99mTcHMPAO. Three of the children had localized cerebral abnormalities on neuroimaging, i.e. right frontotemporal cortical microdysgenesis, left frontotemporal polymicrogyria, and right parietooccipitotemporal porencephalic cyst. Neurophysiologic studies also indicated that a single cerebral hemisphere was predominantly involved, while the other hemisphere and the brainstem were relatively spared, a result compatible with the clinical findings of hemiparesis and hemiconvulsion. In these three patients, administration of carbamazepine was followed by marked improvement of the clinical and electroencephalographic findings. Our results suggest that a combination of noninvasive examinations can distinguish a particular subtype of infantile spasms associated with a localized cerebral abnormality and responsive to carbamazepine.

Effects of compression pressure on physical and chemical stability of tablets containing an anticancer drug TAT-59.

(E)-4-[1-[4-[2-(Dimethylamino)ethoxy]phenyl]-2-(4-isopropyl) phenyl]-1-butenyl]phenyl monophosphate (TAT-59) is a new drug for the treatment of breast cancer. Physical and chemical stability of mixtures of TAT-59 and microcrystalline cellulose (1:9) compressed at 0, 300, 600 and 1400 kg/cm2 was evaluated by determination of water content, porosity and the amount of hydrolysis product, DP-TAT-59, formed. The water contents and porosity of these tablets scarcely changed during 57 d at 25-60 degrees C under 50% relative humidity (RH). The degradation rate of TAT-59 increased with increasing compression pressure as well as temperature. The apparent activation energy and frequency factor were determined from an Arrhenius plot of the degradation rate. Activation energy of these tablets was almost the same, while the frequency factor tended to increase with increasing compression pressure. The porosity and pore sizes in TAT-59-containing tablets decreased with increasing compressive force. We speculated from these observations that the increase in compression pressure decreased the distance and increased the contact area between TAT-59 and microcrystalline cellulose. The proximity between TAT-59 and moisture presented at the surface of microcrystalline cellulose by compression was considered to enhance the degradation of TAT-59.

[Self-expandable metallic stent therapy for inferior vena cava obstruction secondary to malignancy: clinical observations].

To evaluate the self-expandable metallic stent therapy for inferior vena caval obstruction (IVCO) secondary to malignant liver tumors, changes in caval pressure, the symptoms and hemobiochemical values were observed. Among 16 IVCO cases with higher caval pressure than 20 cmH2O at the peripheral caval lumen to the stenosis, nine cases consisting of five extracaval compression cases and four intravenous tumor thrombi cases subjected the stent therapy. Other three subjected radiotherapy and the other four cases inactive supportive care. Immediately after the Z-stent implantation, the averaged caval pressure distal to the stenosis decreased from 27.7 +/- 3.5 cmH2O to 14.7 +/- 2.6 cmH2O. One case developed 8 cmH2O increase of right atrial pressure but no lung edema. The urine excretion volume increased after stent. The decrease in caval pressure correlated with the urine volume of the day after the stenting (gamma = 0.83), symptomatic improvements of leg edema (gamma = 0.68), ascites (gamma v 0.51) and scrotal edema (gamma = 0.70). Five cases showed gradual increase in platelet number. All elevated LDH and elevated fibrinogen value decreased. These changes would suggest physiologic benefits of the IVC stent therapy. Compression cases showed better improvements and courses than the thrombi cases. Two thrombi cases endured severe conditions suspected of triggered by the procedure. Conclusively, the stent therapy to the IVC obstruction was thought to effect hemodynamically and hematobiochemically.

Preparation and application of monoclonal antibodies for a sandwich enzyme-linked immunosorbent assay of the major soybean allergen, Gly m Bd 30K.

In order to obtain probes suitable for the determination of the major soybean allergen, Gly m Bd 30 K, in soybean-related processed foods, we have prepared two monoclonal antibodies, F5 of IgG2a and H6 of IgM, by the fusion of P3U1 myeloma cells with the spleen cells of BALB/c mice immunized with the reductively carboxymethylated allergen (RCM-allergen). The two monoclonal antibodies were shown to be specific to the intact allergen as well as the RCM-allergen and to recognize distinct epitopes on the allergen. Of the monoclonal antibodies, F5 was conjugated with peroxidase. H6 and the labeled F5 were applied as the fixing antibody and the labeled first antibody, respectively, for a direct sandwich enzyme-linked immunosorbent assay of the allergen. When the allergen was extracted from the soybean-related foods with Tris-HCl buffer containing sodium dodecylsulfate and mercaptoethanol, the allergen, Gly m Bd 30 K, was shown to be measured in a range of 5-500 ng in this assay.