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Treatment-required accidental ingestion and risk factors among nursery children with wheat allergy

We introduce a state-of-the-art self-aligned contact (SAC) process and patterning process developed by new patterning technology using Atomic Layer Etch (ALE) and Atomic Layer Deposition towards 5/7nm generation.

A new technology has been developed using the atomic layer etching (ALE) and atomic layer deposition (ALD) concepts. It has been applied to self-aligned contacts (SAC) and patterning processes, for the sub 7 nm technology generation. In the SAC process, ultra-high selectivity of SiO2 etching towards SiN is required, for which we have developed quasi-ALE technique for SiO2 etching. We were able to significantly improve the trade-off between the etching ability of SiO2 on the micro slit portions and SiN selectivity. Quasi-ALE precisely controls the reaction layer thickness of the surface, by controlling the radical flux and ion flux independently, and hence enables etching at lower ion energies (E i < 250 eV). On the other hand, in the patterning processes, the shrinking of critical dimensions (CD) without loading is mandatory. Therefore, we developed a new process flow that combines ALD technique and etching. With this method, we were able to achieve CD shrinking at atomic-layer level precision for various patterns, without causing CD loading. In addition, we were also able to uniformly control the CD shrinkage amount across the whole wafer. This is because this technique takes advantage of the deposition step which is independent of the pattern density and the location on the wafer by self-limited reactions.

It remains unclear whether the number of circulating mast cell progenitors is increased in patients with atopic diseases. Distinct genotypes are reported to affect mast cell/basophil activation.We compared the number and function of mast cell progenitors present in the peripheral blood from donors with normal IgE (IgE400 U/mL) and those with atopic dermatitis accompanied by high serum IgE (IgE5000 U/mL).Purified peripheral blood cells were cultured in serum-free methylcellulose containing stem cell factor (SCF), IL-6 plus IL-3. Fresh methylcellulose containing the cytokines was layered over every 2 weeks. The cultured mast cells were retrieved from the methylcellulose and were functionally analysed.Mast cell colonies were distinguished at 6 weeks of culture as other colony types had been degenerated. The number of mast cell colony-forming cells varied depending on donors and was not significantly increased in peripheral blood from the hyper-IgE atopic patients. A significant inversed correlation was found between the number of mast cells per one colony and the ages of donors. The cultured mast cells derived from atopic patients and those from normal IgE donors equally expressed Fc epsilon RI and released histamine through Fc epsilon RI, although IL-4 priming in vitro markedly enhanced the function of mast cells regardless of donors.These results indicate that the number of circulating mast cell progenitors may be regulated by unknown individual factors unrelated to IgE levels. Mast cell function may be regulated largely by environmental factors, such as IL-4, but not determined by their progenitors' genotypes.

Background: β 2 -Adrenergic agonists are the most widely used bronchodilators for the treatment of asthma. On the other hand, there is concern that excessive use of β 2 -agonists may contribute to the exacerbation of asthma. However, the mechanism of such adverse effects of β 2 -agonists is not completely clear. Objective: The aim of this study was to assess the direct influence of β 2 -agonists on airways by analyzing the effect of a β 2 -agonist, fenoterol, on airway sensitivity in an animal model and on tachykinin neurokinin-2 receptor expression in bovine tracheal smooth muscle. Methods: We performed an acetylcholine challenge test on ovalbumin sensitized guinea pigs that were exposed to daily inhalation of ovalbumin and fenoterol. We also investigated the effects of fenoterol on neurokinin-2 receptor messenger RNA and density with Northern blot analysis and receptor binding assay. Result: The increase of airway responsiveness and the decrease of β 2 -adrenergic receptors were found in guinea pigs that were treated with fenoterol. There were time- and dose-dependent increases of neurokinin-2 receptor mRNA and of density in tracheal smooth muscle that was treated with fenoterol. Conclusion: This increased airway responsiveness, increased neurokinin-2 receptor expression, and decreased β 2 -adrenergic receptor density may be relevant to asthma exacerbation. (J Allergy Clin Immunol 2000;106:S104-8.)

Histamine, released from activated mast cells, causes bronchoconstriction mediated by H(1) receptors, whereas beta(2)-agonists are widely used for the relief of bronchoconstriction. In this study, we examined the effects of the beta(2)-adrenoceptor agonist, fenoterol, on the expression of H(1) receptors at the mRNA and protein levels, and functional responses. Incubation of bovine tracheal smooth muscle with fenoterol (10(-7) M) for 2 h increased H(1) receptor mRNA (maximum approximately 190%). The number of H(1) receptors was increased after 12 and 18 h without any change in binding affinity. In the contraction experiments, the concentration-response curves for histamine-induced contraction were shifted significantly to the left after 18-h exposure to fenoterol, consistent with the increase in receptor number. The fenoterol-induced increase in H(1) receptor mRNA was concentration-dependent and was abolished by propranolol and ICI 118551, but not by CGP 20712A, indicating that fenoterol acts via beta(2)-adrenoceptors. These effects were mimicked by other cAMP-elevating agents forskolin and prostaglandin E(2), and by the stable cAMP analog 8-bromo-cAMP. Cycloheximide alone produced superinduction of H(1) receptor mRNA and augmented the fenoterol-induced increase in H(1) receptor mRNA. Fenoterol increased both the stability and the transcription rate of H(1) receptor mRNA. Pretreatment with dexamethasone did not prevent fenoterol-induced up-regulation of H(1) receptor mRNA. Thus, fenoterol increases the expression of airway smooth muscle H(1) receptors via activation of the cAMP system through increased gene transcription and mRNA stability. This mechanism may be involved in the adverse responses encountered with the clinical use of short-acting beta(2)-agonists.

Serum glutamic-oxaloacetic transaminase (GOT) levels were determined in 214 infants (133 males and 81 females) with atopic dermatitis during their first visit to the Department of Allergy, National Children's Hospital, Tokyo, Japan. Compared with the normal hospital range, their levels were found to be significantly higher, a tendency which was more conspicuous in lower age groups. We carried out a 13C-methacetin breath test (MBT), administering the stable-isotope-labeled compound to 11 children with higher serum GOT values and 5 within the normal range to investigate hepatic metabolism of methacetin in infants with atopic dermatitis. 13C-methacetin was given orally, and the 13CO2 level in the breath was determined immediately before and after administration, by mass spectrometry. Compared to the normal controls, the atopic infants demonstrated significantly lower 13CO2 peak excretion and delayed peak time. The clearance rate of 13CO2 was also decreased. These results suggest some relationship between atopic dermatitis and liver function in infants.

Sinusitis is common in children with allergic diseases, and the relationship between sinusitis and reactive airway diseases involving asthma has been reported. Most pediatricians and physicians base their diagnosis of sinusitis on findings from plain radiographs of the sinuses, especially Waters projection radiographs. We compared the diagnoses made by 11 pediatric allergists using 56 Waters projection radiographs with transverse CT findings. The ratio for the two diagnosis being consistent (normal plain radiographic findings and normal CT findings, or abnormal plain radiographic findings and abnormal CT findings) was approximately 60%. Sinusitis in children is often misdiagnosed on the basis of findings from Waters projection radiographs alone. Therefore, the use of CT findings for the diagnosis of sinusitis together with Waters projection radiographs is recommended.

The function of the hypothalamic-pituitary-adrenal (HPA) axis was investigated in nine asthmatic children treated by inhalation of beclomethasone dipropionate (300 micrograms/day) for 12 weeks. The trial was designed as a prospective study. The 24-hour urinary free cortisol, serum cortisol, response to ACTH, and nocturnal serum cortisol followed by 20-minute sampling were measured before and after the study period. No significant changes were found, suggesting that inhaled beclomethasone, in a daily dose of 300 micrograms, does not cause suppression of the HPA axis.

We present a case of wheat-dependent exercise-induced anaphylaxis, in which pretreatment with sodium bicarbonate inhibited reappearance of anaphylactic symptoms following wheat and exercise provocation. Decrease in blood pH relative to elevation in plasma histamine levels was also inhibited. These results suggest possible efficacy of pretreatment with sodium bicarbonate as a preventive measure in patients with exercise-induced anaphylaxis.

A 64-year-old man was referred to our hospital because of little improvement of hemoptysis lasting three days after drug therapy. A chest roentgenogram and fiberoptic bronchoscopic examination performed on the second hospital day when the patient experienced a massive hemoptysis of about 2,000 ml revealed arterial bleeding from the left upper lobe. Even after extensive embolizations of the left upper bronchial, the first, second, third and 4th intercostal arteries, the patient's hemoptysis did not improve. On the 73rd hospital day the patient underwent left upper lobectomy. Macroscopic and microscopic examinations in the resected specimen revealed lung injury due to a torn segment of pleural calcification after tuberculous pleuritis, resulting in the massive hemoptysis. Although physicians encounter many patients complaining of hemoptysis and/or hemosputum, this case is considered to be very rare.

We present a case of exercise-induced anaphylaxis with improvement following the removal of dental amalgam. Although her symptoms were unresponsive to various kinds of therapy until removal of the amalgam, her symptoms related to exercise improved remarkably after the removal. The increase in plasma histamine levels for exercise provocation test also improved. This suggests that sensitivity to metals might cause exercise-induced asthma in some patients.

The role of intestinal flora in the production of anorexigenic substance was investigated. Proteus mirabilis ( P . mirabilis ) and Escherichia coli ( E. coli ) were found to produce an anorexigenic substance, while Enterococcus faecalis ( E. faecalis , type 1 and 2) and Staphylococcus intermedius ( S. intermedius ) did not. The anorexigenic substance was purified and was detected as, a single though broad band by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity of the final form of the purified substance was 120 units/mg carbohydrate. The substance contained no protein residue and appeared to be a lipopolysaccharide. The evidence that intestinal flora produces an anorexigenic substance leads to an interesting assumption that the intestinal flora may be responsible for regulating food intake.

A DNA-binding protein with a molecular weight of 64,000 (64DP) has been found in elevated concentrations in the serum of patients with malignant diseases. 64DP has been found to be different from any previously described tumor markers, such as AFP, CEA, and C3DP. The results obtained with column SDS - polyacrylamide gel electrophoresis are as follows. 64DP is normally present in the serum at the concentration of 48 +/- 26 micrograms/ml (n = 49). The mean value of 64DP in 96 patients with untreated malignant disease was 167 +/- 67 micrograms/ml, whereas in 41 patients with nonneoplastic disease it was 61 +/- 23 micrograms/ml. No organ or tissue specificity seems to be required for malignant neoplasms to result in elevated serum 64DP concentrations. The clinical stage of the disease does no have much effect on the levels of 64DP and, even with histologically proven early lesions, the patients showed elevated levels of serum 64DP. Following successful surgical resection, 64DP tends to fall toward the normal value. Purification of 64DP was performed and its physicochemical properties have been defined. 64DP is a glycoprotein present in the alpha1-globulin fraction and it is rich in leucine, glutamic acid, and aspartic acid in its amino acid composition. More recently, using purified 64DP preparation, a single radial immunodiffusion assay system has been developed. According to the results of this method, the diagnostic accuracy of 64DP appears to be somewhat low and even marginal for certain types of commonly encountered surgical malignancies. Studies concerning the discrepancy between the results obtained by the two assay methods are now in progress.

Inhibition studies of crystallized rat liver argininosuccinate synthetase [EC 6.3.4.5] are described. 1. L-Argininosuccinate, L-histidine, and L-tryptophan inhibited the enzyme activity at saturating amounts of the substrates. 2. L-Norvaline, L-argininosuccinate, L-arginine, L-isoleucine, and L-valine competitively inhibited the enzyme activity at a low concentration of L-citrulline, with Ki values of 1.3 x 10(4) M, 2.5 X 10(-4) M, 6.7 X 10(-4) M, 6.3 X 10(-4) M, and 6.0 x 10(-4) M, respectively. 3. L-Argininosuccinate and L-arginine competitively inhibited the enzyme activity at a low concentration of L-aspartate, with Ki values of 9.5 x 10(-4) M and 1.2 x 10(-3) M, respectively. 4. The modes of inhibition by L-histidine were mixed-noncompetitive, uncompetitive, and noncompetitive types with respect to L-citrulline, L-aspartate, and ATP, respectively. 5. When the enzyme was preincubated with L-citrulline, the enzyme activity was slightly increased in the presence of a low concentration of L-histidine in the assay mixture. 6. The conformation of the enzyme was markedly changed by the addition of L-histidine as judged from the CD spectrum. This change was partially reversed by incubation with L-citrulline.

The biosynthesis of urea is regulated mainly by two factors, the amounts of urea cycle enzymes and the concentrations of acetyl-glutamate and ornithine. Schimke1 pointed out that the contents of all the urea cycle enzymes in the liver were directly proportional to the daily consumption of protein, then the activities of urea cycle enzymes are an important regulatory factor of the urea cycle. On the other hand, other investigators2–4 reported that the concentration of acetylglutamate, an allosteric activator of carbamylphosphate synthesis, and of ornithine, the rate limiting intermediate, changed under various dietary conditions and suggested that these amino acids play a role in the regulation of urea synthesis. We reported that ornithine and acetylglutamate play a more important role in the regulation of urea synthesis especially shortly after the dietary change. In the liver of rats subjected to acute dietary transitions from high to low protein or vice versa, the concentrations of ornithine and acetylglutamate changed greater and prior to the activity changes of urea cycle enzymes. The rate of urea synthesis from ammonium salt as a substrate were greatly changed in the perfused liver and correlated with the changes in the concentration of ornithine in the liver after the dietary changes5. Arginine derived from dietary protein is thought to be the main source of ornithine and also the cause of changes in acetylglutamate3–5. However, other factors must be involved in the regulation of the concentration of ornithine, since the concentration of orni-thine as well as acetylglutamate increased after the intraperitoneal injection of the ammonium salt without altering either arginine or protein input6.

This study was carried out to elucidate the interaction of alpha-1-antichymotrypsin (ACT) with the immune reaction induced by interleukin-2 (IL-2). ACT, isolated from human sera and determined to be a single glycoprotein, was added to human peripheral T-cell cultured with IL-2. It was found that ACT effectively suppressed T-cell activation induced by IL-2, but the suppressive effect of ACT was inversely proportional to the concentration of IL-2. A similar effect was found when T-cells were stimulated with PHA. ACT had a suppressive effect when suboptimal concentrations of PHA were used, however this suppressive effect could not be found at the optimal PHA concentration. These findings suggest that ACT may be a regulatory agent in immune reactivity where low concentrations of IL-2 are present. Even under normal conditions, an individual is surrounded and invaded by numerous foreign elements and it may be that small amounts of IL-2 are constantly produced, which would indefinitely trigger chained immune reactions if there were no regulatory agent like ACT.

A DNA-binding protein with a molecular weight of 64,000 (64-DP) has been found in elevated concentrations in the serum of patients with malignant diseases, and a quantitative assay system (column-SDS disc electrophoresis method) for it has been developed. The average serum 64-DP level in 49 normal controls was 48 +/- 26 microgram/ml, whereas in 96 patients with untreated malignant disease it was 167 +/- 67 microgram/ml. Sera from patients with non-neoplastic diseases showed normal or only slightly elevated 64-DP concentrations. No organ or tissue specificity seems to be required for malignant growths to result in elevated serum 64-DP concentration. Of particular interest was the finding that all 11 patients with early gastric carcinoma and two patients with early esophageal carcinoma, all having histologically proven pT1 lesions, showed elevated levels of serum 64-DP. Following successful surgical resection and/or chemotherapy, 64-DP tends to fall toward the normal value. Although confirmation must await the results of larger clinical trials, our preliminary results strongly suggest that 64-DP evaluated by the column-SDS disc electrophoresis method may prove to be a useful tumor marker in patients with a wide variety of malignancies.

The relationship between chymotrypsin-inhibitory and immunoenhancing activity of alpha-1-antichymotrypsin was studied. alpha-1-Antichymotrypsin was treated at 50 degrees C, 55 degrees C or 60 degrees C for 15 min. It was found that antichymotryptic activity was reduced by half when alpha-1-antichymotrypsin was heated at 55 degrees C and was not detected at all when heating was carried out at 60 degrees C. alpha-1-Antichymotrypsin which was heated at 60 degrees C did not form a complex with chymotrypsin, but became a substrate for chymotrypsin. The effect of native and heated alpha-1-antichymotrypsin on antibody response was studied in mice. alpha-1-Antichymotrypsin increased the number of anti-sheep erythrocytes antibody producing cells even when it was heated at 60 degrees C. Circular dichroism and single radial immunodiffusion were used to detect conformational changes. Circular dichroism in the region of side chain absorption showed that the intensities of the spectra at 296, 284, and 265 nm decreased with a rise in temperature from 50 to 60 degrees C. In single radial immunodiffusion analysis, alpha-1-antichymotrypsin did not form a halo after being heated at 60 degrees C. In conclusion, when alpha-1-antichymotrypsin was heated at 60 degrees C, the immunoenhancing activity remained intact while the antichymotryptic activity was lost with the conformational change.

The effect of alpha-1-antichymotrypsin (ACT), which is known as an efficient serum protease inhibitor and is detected in tumor cell nuclei, on DNA synthesis was studied. ACT inhibited the activity of DNA polymerase alpha purified from human stomach adenocarcinoma. Other human serum proteins including serum albumin, alpha-1-acidglycoprotein, alpha-1-antitrypsin, and immunoglobulin G, as well as other protease inhibitors, such as leupeptin, pepstatin, PMSF and chymostatin, did not affect the activity of DNA polymerase alpha. It was therefore concluded that the inhibitory action of ACT on DNA polymerase alpha was direct phenomenon unrelated to its protease inhibitory activity. Furthermore, the effect of ACT on DNA synthesis was also studied using lysolecithin-permeabilized cultured human stomach carcinoma cells. ACT added in the medium inhibited DNA synthesis and the degree of inhibition depended on incubation time. It was proportional to ACT concentration and the concentration of ACT required for 50% inhibition was 0.8 mg/ml.

This paper describes an investigation into the effect of alpha-1-antichymotrypsin (ACT) on DNA primase. DNA primase was partially purified from human stomach carcinoma cells. It was found that poly(dC)-dependent DNa primase activity was inhibited by ACT and the inhibition was proportional to the concentration of the inhibitor. The inhibitory effect of ACT remained even after ACT lost most of its chymotrypsin-inhibitory activity by heat treatment. Poly(dT)-dependent primase activity was enhanced by the presence of ACT. The enhancement was effective up to a concentration of 1mg/ml.

Human stomach adenocarcinomas containing alpha-1-antichymotrypsin (ACT) in their cell nuclei were transplanted into nude mice. The presence of ACT was monitored using an immunohistochemical technique with horseradish peroxidase-labeled rabbit anti-ACT Fab' as well as single radial immunodiffusion. Two weeks after transplantation, ACT could be found neither in transplanted carcinoma cells nor in the sera of carcinoma-bearing nude mice. However, if human ACT was injected i.v., it could be detected in the transplanted carcinoma cell nuclei 2 h after injection. The ACT was detected immunohistochemically and was confirmed by biochemical fractionation using 125I-labeled ACT. On the other hand, the amount of ACT production was not sufficient to indicate biosynthesis. These results demonstrated that ACT detected in stomach carcinoma cell nuclei was not synthesized in carcinoma cells but was incorporated from the blood circulation.

Sialidase activity of peripheral mononuclear cells, which are mostly lymphocytes, was found to be increased by lectin stimulations in in vitro experiments, but this induction was suppressed in the presence of 50 mM ethanol. This increasing change and its suppressive effect by lectins and ethanol were parallel to blastogenic change of the cells determined by 3H-thymidine uptake. In in vivo, sialidase activity of peripheral MNC prepared from patients with alcoholic liver disease was found to be decreased or not increased in 50% of the cases, in contrast to the marked increase of the activity in non-alcoholic liver diseases observed in our previous study.

alpha 1-Antichymotrypsin (ACT), which is known as an efficient serum protease inhibitor and is detected in tumor cell nuclei, was found to inhibit the activity of DNA polymerase alpha purified from human stomach adenocarcinoma. The concentration of ACT required for 50% inhibition was 1.0 mg/ml and the manner of its inhibition showed the partially competitive relationship between ACT and DNA in the assay system. Furthermore the removal of ACT by anti-ACT antibody lost its antichymotryptic and anti-DNA polymerase activities in parallel. On the other hand, it did not inhibit the activity of human DNA polymerase beta. Other human serum proteins including serum albumin, alpha 1-acid glycoprotein, alpha 1-antitrypsin, and immunoglobulin G as well as other protease inhibitors such as leupeptin, pepstatin, phenylmethylsulfonyl fluoride, and chymostatin did not affect the activity of DNA polymerase alpha. Furthermore ACT heated at 60 degrees C did not inhibit DNA polymerase alpha, although it could still bind to DNA as well as native ACT. It was therefore concluded that the inhibitory action of ACT on DNA polymerase alpha was a direct phenomenon unrelated to its protease inhibitory or DNA binding activities.

Properties of argininosuccinate synthetases (ASS), including immunological properties and heat stability, from young and old rats were quite similar. However the degradation rates of ASS from young and old rats were found to be different as shown in the experiment using the double labeling technique. Three forms of this enzyme from young and old rat livers were separated by DEAE-Sephadex A-50 column chromatography, and they were called ASS 1, 2 and 3 in order of elution. The degradation rate of ASS 1 from young and old rat livers was very similar, but the rate of degradation of ASS 2 form old rat livers was greater than that from young rat livers.

The levels of alpha 1-antichymotrypsin (ACT) was monitored in patients who underwent bone marrow transplantation. Seven received HLA-identical sibling bone marrow grafts, two received transplants from twins and one was given HLA-nonidentical marrow from his father. A dramatic increase of ACT was observed in all patients who developed graft-versus-host disease (GvHD). ACT did not rise at all in the case of patients who received marrow from twins, even in a patient who was given three transplants from the same donor. The patient transplanted from his father died from GvHD and the increase of ACT was the greatest fluctuation measured.

A DNA binding protein with a molecular weight of 64,000, designated 64DP, has been purified and characterized. This protein was isolated from adult human pooled serum by DEAE Sephadex column Chromatography, DNA cellulose affinity column chromatography, ammonium sulfate fractionation and Sephadex G-150 gel filtration. The final preparation of 64DP was homogeneous, as judged from polyacrylamide gel electrophoresis with or without sodium dodecyl sulfate and sedimentation experiments. Physicochemical and immunochemical properties of this protein were very similar or identical to those of alpha-1-antichymotrypsin with some differences in electric mobility and th pattern of isoelectric focusing. Furthermore, the general properties of 64DP from various sera were practically similar with the exception that isolectric focusing analysis showed microheterogeneity among 64DP purified from various sera.

The cellular localization of alpha-1-antichymotrypsin (alpha 1-ACT) was studied immunohistochemically using rabbit HRP-labeled Fab' against human alpha 1-ACT. alpha 1-ACT was found in cell nuclei of carcinomas of the stomach, liver, breast, pancreas and leiomyosarcoma and in cell nuclei of lymphoid cells infiltrated into the stomach carcinoma mass. alpha 1-ACT was not found in carcinoma cells of the colon, uterus, rectum or esophagus, or in lymphoid cells infiltrated into the rectal carcinoma mass or into inflammatory regions such as gastric ulcers or appendicitis. Further, alpha 1-ACT was not found in normal cells around the carcinoma mass or in normal tissues.

A DNA binding protein in human serum was purified and identified. This protein was isolated from pooled human serum by DEAE Sephadex column chromatography, DNA cellulose affinity column chromatography, Sephadex G-200 gel filtration and Sepharose 6B gel filtration. It was found that the purified protein consisted of two subunits, one with a molecular weight of 80,000 and the other 25,000, evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified protein was in fact IgM determined by physicochemical and immunochemical characteristics.

Incorporation of alpha-1-antichymotrypsin (ACT) into human stomach adenocarcinoma cell nuclei and the effect of ACT on DNA primase from the same carcinoma cells were studied. ACT or [125I]-ACT were observed in carcinoma cell nuclei and high specific radioactivity was detected in washed nuclear fraction when 0.4 mg of ACT or [125I] ACT (8 x 10(7) cpm) was intravenously injected into carcinoma bearing nude mice 2 h before killing. The molecular weight of radioactivity presented in cell nuclei was same as the intact ACT on SDS-polyacrylamide gel electrophoresis. ACT inhibited DNA primase activity and this inhibiting activity was stable than its chymotrypsin inhibiting activity. The results presented here show ACT is incorporated into carcinoma cell nuclei without modification of its molecular weight and may inhibit DNA primase activity.