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我が国の喘息死は小児，成人共に減少傾向にあり，喘息死亡率も減少し概ね低率で安定しつつある1）～3）．日本小児アレルギー学会・喘息死委員会宛に2010年10月までに，気管支喘息があって死亡し登録された症例は233例で，対象外を除いた206例について解析した．2010年には2名の登録があった．男女比は1997年以前98/63（1.6：1），1998年以降28/17（1.6：1）で，変化はみられなかった．不明・未記入を除いて死亡前1年間の重症度をみると1997年以前は軽症26％，中等症30％，重症44％，1998年以降は軽症35％，中等症27％，重症38％で，重症度に大きな変化は見られなかった．死亡年齢と死亡場所の関係をみると自施設で死亡した例は0～6歳73％，7～12歳55％，13歳以上では38％と加齢に伴い減少傾向が認められた．逆に自施設以外の場所での死亡例が年長になるほど増加する傾向にあり，学校内あるいは養護学校，下校時など学校が関与する症例が認められた．発作重症化に関わる要因として，入院歴が52％に認められたが，意識障害を来たすほどの重症発作，挿管，isoproterenolによる治療の既往は少なかった．喘息死に関与した要因では，予期できない急激な悪化，適切な受診時期の遅れが最も多かった．適切な受診時期の遅れを来たした要因として，患者・家族による判断の誤りが多く，短時間作動性β2刺激薬の加圧噴霧式定量吸入器（pMDI）やモーターネブライザー（MoNe）への過度依存，怠薬がそれに次いだ．薬物療法について，1997年以前の死亡例を1998年以降と比較すると，キサンチン製剤とβ2刺激薬の内服，自宅でのMoNe吸入，pMDIは減少し，吸入ステロイド（ICS）は増加傾向にあるものの38％に留まっていた．また，β2刺激薬貼付剤やLABA（long acting β2agonist）の使用が新たに認められるようになった．死亡前1年，1ヶ月間の重症度別薬剤使用状況を解析した結果，吸入ステロイド，ステロイド内服とも，使用していない症例が少なからず報告されており，死亡前にコントローラーとして充分に使用されていなかった可能性がうかがえた．また，急性発作増悪時にSABA（short acting β2agonist）定量噴霧式吸入が充分に行われていなかった可能性も考えられた．

The role of complement in the pathogenesis of marked thrombocytopenia induced in rats by Erysipelothrix rhusiopathiae (E. rhusiopathiae) was examined. In cobra venom factor (CoF)-treated rats thrombocytopenia was not induced by the bacterium. The content of 5-hydroxytryptamine (5-HT) in platelets was decreased significantly after inoculation only in untreated rats. E. rhusiopathiae could bind C3 and generate platelet-bacteria aggregation when incubated in the plasma diluted with veronal-buffered saline containing calcium and magnesium (VBS++) or gelatin-VBS containing magnesium and ethyleneglycol tetra-acetic acid (GVB-Mg-EGTA), but not when incubated in GVB-ethylenediamine tetra-acetic acid (GVB-EDTA) diluted plasma or in CoF-treated or anti rat C3-treated plasma. When platelets were preincubated with activated zymosan, no bacteria could bind to platelets. From the results, we speculate that the alternative complement pathway, activated by E. rhusiopathiae, appears to mediate the formation of platelet-bacterial aggregations that may accelerate the removal of platelets from circulating blood.

This study is designed to investigate whether apoptosis occurs in vivo in pediatric patients with acute leukemia during induction therapy. When patients with common acute lymphoblastic leukemia (cALL) and acute myeloblastic leukemia (AML) were treated with prednisolone (60 mg/m2/day, p.o. or i.v.) and etoposide (150 mg/m2/day, i.v.), respectively, the blast cell counts fell to below 30% and 5%, respectively, in 1 week. However, during this cytoreduction phase, neither morphologically apoptotic cells nor fragmentation of DNA derived from peripheral blast cells were detected at any preparations. On the other hand, cALL but not AML cells spontaneously undergo apoptosis following their culture in vitro. The addition of autologous serum instead of fetal calf serum substantially prevented apoptosis from occurring spontaneously in cALL cells. When cALL and AML cells freshly obtained from patients before therapy were treated in vitro with 10 mumol/l prednisolone and 20 micrograms/ml etoposide, respectively, these cells underwent apoptosis within 6 hours, as determined by a morphological and DNA fragmentation assay. These in vivo and in vitro findings suggest that, although anticancer drugs may induce apoptosis in vivo, these apoptotic cells cannot be detected due to their rapid removal from the circulation.

Sequential CT studies were performed in 24 patients with infantile spasms, before, during and after ACTH therapy, in order to analyze the relationship between the dose of ACTH and changes in CT scan findings. In six cases CT scans did not return to their original state after ACTH therapy. Two illustrative cases are reported. There was a good correlation between the total dose of ACTH and the percentage decrease of brain parenchyma. It is concluded that 'cerebral atrophy' on CT scans may represent an accumulative effect of ACTH and that ACTH should be given with the utmost discretion, and in as low a dose as possible.

Asymmetrical head-turning and its effects on subsequent functional lateralities and postural asymmetries were examined in 44 relatively low-risk preterm and 53 fullterm infants. Persisting head-turning to the right was observed more often among preterm infants, as was an asymmetrical skull with flattened right occiput and head-turning to the right. Asymmetrical trunk posture was present in 20 preterm and four fullterm infants. At nine months right-hand preference was more marked in preterm infants, and hand preference was well coincident with head-turning. Changes in hand preference occurred more often among fullterm infants. Asymmetrical gait seemed to correlate with head-turning in preterm infants.

The cytotoxicity and DNA lesions induced by methotrexate (MTX) were compared in wild-type, hypoxanthine-guanine phosphoribosyltransferase-deficient (HGPRT-) and thymidine-kinase-deficient (TK-) HL-60 cells. TK- and HGPRT- cells were approximately 10 and 3 times more sensitive to MTX than wild-type cells, respectively. Following incubation with 2 microM MTX for 16 hr, TK- cells showed a significantly higher number of DNA strand breaks. Concomitantly, DNA fragmentation at the nucleosomal linker region was detected more prominently in TK- cells. Although MTX tended to decrease TTP pools similarly in all 3 cells types, the initial TTP level in TK- cells was only about one-fifth of that found in the wild type. These results indicate that the thymidine salvage pathway has a pivotal role in mediating MTX-induced toxicity and DNA lesions.

Changes in nasopharyngeal bacterial flora in adults with acute upper respiratory tract infection on administration of antimicrobial agents were investigated, and how these changes contrasted with those in children. Many patients with acute sinusitis due to allergies, and patients with malignancy and diabetes mellitus were included in the investigation. The detection rates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis, the major bacteria of acute otitis media (AOM), were 22%, 10%, and 7% respectively, which were significantly lower than those for children. Gram stain examination of nasopharyngeal swab samples showed a significant relation between leukocyte infiltration and the detection amount of S. pneumoniae (P = 0.0086). A significant relation (P = 0.0134) was also observed when H. influenzae was simultaneously detected. No significant change in the three major AOM bacteria present in nasopharyngeal bacterial flora after administration of antimicrobial agents was observed. However, all S. pneumoniae and H. influenzae detected after antimicrobial agent administration had the beta-lactam-resistance gene. It was observed that a significant improvement in leukocyte infiltration occurred 6 to 10 days after antimicrobial agent administration. In contrast, a significant improvement in children was observed at 2 to 5 days. In the adult subjects, this improvement was probably due to spontaneous remission rather than the effect of the antimicrobial agents. Although investigation of the long-term administration of antimicrobial agents was also conducted, its benefits for the patients were not elucidated.

Background Hydrofluoroalkane-134a (HFA) has been shown to be a safe replacement for chlorofluorocarbons (CFCs) as a pharmaceutical propellant, with the advantage that it has no ozone-depleting potential. This is the first report of the pharmacokinetics of beclomethasone dipropionate (BDP) delivered from a pressurized solution formulation using an HFA propellant system (HFA-BDP) in Japanese children with bronchial asthma. Methods Plasma concentrations of beclomethasone 17-monopropionate (17-BMP), a major metabolite of BDP, following an inhaled dose of HFA-BDP (200 μg as four inhalations from 50 μg/actuation) in five Japanese children with bronchial asthma were quantified and analyzed by a non-compartmental analysis to obtain pharmacokinetic parameters. Results The area under the concentration-time curve from time zero to the last quantifiable time (AUC 0-t ) was 1659 ± 850 pg • h/mL (arithmetic mean ± standard deviation (SD)), the maximum concentration observed (C ma x) was 825 ± 453 pg/mL and the apparent elimination half-life (t 1/2 ) was 2.1 ± 0.7 hours. The time to reach C max (T max ) was 0.5 hours in all patients. No special relationship was observed between these parameters and age or body weight. These parameters were compared with the previously reported parameters of American children with bronchial asthma. The Japanese/American ratio of the geometric means of each parameter was 1.36 for AUC 0-t , 1.04 for C max and 1.4 for t 1/2 . The median of T max was 0.5 hours in American patients as well as Japanese patients. Conclusions The pharmacokinetics of HFA-BDP in Japanese children with bronchial asthma are reported for the first time and a similarity to those in American children is suggested.

This report focuses on changes in the nasopharyngeal bacterial flora before and after administration of antimicrobial agents in 172 cases of acute upper respiratory infection in patients aged 6 years or younger. The antimicrobial agents administered were amoxicillin (AMPC) (34%), clavulanic acid/amoxicillin compound (11%), cefditren pivoxil (CDTR-PI) (43%), and others (12%). Changes in nasopharyngeal bacterial flora were investigated with reexaminations conducted after 2-5 days (day 2-5 subgroup), 6-10 days (day 6-10 subgroup), and 11 days and thereafter. There was a significant reduction in the Streptococcus pneumoniae detected in the group administered AMPC (AMPC group) in the day 2-5 subgroup and the day 6-10 subgroup. There was also a significant decrease in H. influenzae in the group administered CDTR-PI (CDTR-PI group) in the day 2-5 subgroup. From this it was inferred that for the most part significant changes in infectious nasopharyngeal bacteria occurred in the day 2-5 subgroups. However, a significant improvement in the degree of inflammation, as indicated by leukocyte infiltration images for the AMPC group, was observed in the day 2-5 subgroup, and for the CDTR-PI group in the day 6-10 subgroup. On the other hand, in both the antimicrobial agent groups, S. pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis were newly detected at reexamination. Furthermore, a difference in the incidence of these bacteria was observed between the 2 antimicrobial agent groups. It was suggested that such phenomena related to the survival of resistant strains or a recurrence otitis media.

In our first report, we investigated nasopharyngeal bacterial flora related to penicillin-resistant Streptococcus pneumoniae (PRSP) and β-lactamase-negative ampicillin-resistant Haemophilus influenzae (BLNAR) and their relation to acute upper respiratory tract infection (AURTI). This report analyzes the results of a study of nasopharyngeal bacterial flora before the administration of antimicrobial agents in 172 AURTI patients aged 6 years or younger. In addition to Gram staining, microscopic observation, and culturing, a polymerase chain reaction (PCR) method was used to identify PRSP (gPRSP) and BLNAR (gBLNAR) drug-resistant genes. Of the patients analyzed, 90% had acute otitis media (AOM) and were aged 2 years or younger. The antimicrobial agents administered were amoxicillin (34%), clavulanic acid/amoxicillin (11%), cefditren pivoxil (CDTR-PI) (43%), and others (12%). This was particularly true for patients administered CDTR-PI, among whom there were many who had already suffered one or more episodes of AOM by the age of 1 year or younger, and many in which gPRSP were detected (P < 0.01). There was a significant relation between the degree of nasopharyngeal inflammation indicated by leukocyte infiltration images and the amount of S. pneumoniae and H. influenzae detected, which are the main pathogenic bacteria causing AOM (P < 0.01). In addition to leukocyte infiltration images, there were cases in which shedding of ciliated cells was observed and/or giant monocytic cells. Both nasopharyngeal leukocyte infiltration images and/or shed cell findings observed in infant AURTI cases are important indices for the prompt detection of gPRSP and/or gBLNAR and appropriate doses of antimicrobial agents.

Background This is the first report about the pharmacokinetics (PK) of pranlukast in children. The aim of the present study was to assess the PK parameters of pranlukast in children and to compare them with those in adults. Methods Six healthy adult male volunteers and 22 children with bronchial asthma at 3–14 years of age were enrolled in the study. Both 225 and 112.5 mg pranlukast hydrate dry syrup was administered orally to adults, whereas 3.5 mg/kg pranlukast hydrate dry syrup was given to children. Blood samples were obtained at approximately 20 time points per adult (n = 121) and at two or three time points per child (n = 54). Population PK analysis was performed using NONMEM (Globomax, Hanover, MD, USA). The concentration-time-course of pranlukast was described by using a one-compartment model with first-order absorption. The robustness of the final model was evaluated using 200 bootstrap samples. Results Apparent clearance (CL/F) was 1.81 and 1.14 L/h per kg in children and adults, respectively. According to subgrouping of children, no significant difference was observed in CL/F between infants (3–6 years of age) and schoolchildren (7–14 years of age). The interindividual variability of CL/F accounted for 48.7%. The additive and proportional residual variability was 7.33 ng/mL and 73.8%, respectively. All fixed effect parameters fell within 10% of the bootstrapped mean. Conclusions Compared with adults, children showed a higher CL/F and more rapid elimination after ingestion of pranlukast hydrate dry syrup. However, no significant variation was seen in CL/F between infants and schoolchildren.

A retrospective survey of a family history of allergy employing a special questionnaire for children was performed. The survey included 1-year-old (n = 267) and 6-year-old children (n = 410) with allergies as well as 1-year-old (n = 313) and 6-year-old children (n = 329) without allergies. An 'Allergy Risk Score' (ARS) for each subject was calculated according to the history of allergy in three family members: father, mother and a sibling. Each family member was scored as 2.0 (overt history of allergies), 1.0 (provable), 0.5 (possible) or 0 (absent), and the ARS of the subjects was calculated as the total of the family members' scores. The ARS of children in the allergy groups was statistically higher than that of the control groups in both age groups. The ARS increased with an increase in the odds ratio (an approximate value of the risk), especially in 6-year-old children. An ARS calculated on the basis of family history could be a useful and practical index for estimating the risk of allergy development in children, and may be helpful in predicting and preventing allergies in infants and children.

A questionnaire on the treatment of asthma was sent to 586 physicians. They consisted of specialists authorized by the Japanese Society of Allergology and councillors of the society who were treating patients with bronchial asthma. Of the total of 306 (52%) respondents, 241 replied to questions relating to adult asthma and 129 to questions relating to childhood asthma (including duplicate replies). For acute treatment, methods most commonly selected by physicians were, in increasing order of popularity, for adults: parenteral aminophylline, oxygen inhalation, parenteral steroids and parenteral adrenaline; for schoolchildren (6–16 years): parenteral aminophylline, inhaled β-stimulant, oxygen inhalation, inhaled β-stimulant + disodium cromoglycate (DSCG), parenteral steroids; and for infants (≤ 5 years): inhaled β-stimulant, parenteral aminophylline, oxygen inhalation, inhaled β-stimulant + DSCG and parenteral steroids. For maintenance treatment, methods most commonly selected by physicians were, in increasing order of popularity, for adults: oral administration of sustained-release theophylline, inhaled steroids and DSCG inhalation; for schoolchildren (6–16 years): DSCG inhalation, oral administration of sustained-release theophylline, oral administration of antiallergic agent preparation and (β-stimulant + DSCG inhalant; and for infants: DSCG inhalation, oral administration of sustained-release theophylline, oral administration of antiallergicagent preparation and β-stimulant + DSCG inhalant. The questionnaire results clearly showed that different drugs were selected for the treatment of asthma in adults, schoolchildren and infants.

To clarify the effect of the diet in infancy on the development of atopic dermatitis and the production of IgE antibody against mite, 96 babies received varieties of diet were evaluated retrospectively. They were divided into 5 groups. The infants restricted egg from early infancy were divided into 2 groups (group A: showed no food allergy, n=10, group A': showed allergy against some foods, n=14). Eighteen were excluded food (s) that exacerbated symptoms (group B). Thirteen could not avoided the food (s) which worsened their clinical manifestations (group C). Infants showed no exacerbation after ingestion of food were fed unrestricted diet (group D, n=29). The other12 were control infants (group E).We could not find any difference in the development of atopic dermatitis between group A and D, and between grup A' and B. The dermatitis was significantly severe in group C. About 39% of positive RAST related with the clinical symptoms. The incidence of food allergy was high in infants with RAST scores 3 or more and with more than 3 kinds of positive RAST. The production of IgE antibody against mite was also most frequently detected in those with more than 3 kinds of positive RAST. The incidence of mite allergy was significantly high in group C, and the incidence in group Bwas higher than that in group A'.In conclusion, the eliminating diet in infancy is effective for the prevention of the development of atopic dermatitis and the production of IgE antibody against mite in children who showed exacerbation of symptoms after ingestion of the food. The ingestion of egg from early infancy was supposed to affect partly the developement of allgic diseasein future.

A questionnaire on asthma management was sent to 586 physicians, consisting of specialists authorized by the Japanese Society of Allergology and councillors of the Society, who were treating patients with bronchial asthma. Of the total of 306 (52%) respondents in November 1993, 241 replied to questions relating to adult asthma and 129 to questions relating to childhood asthma (including duplicate replies). Responses to the questionnaire on the diagnosis, patient education and management of asthma indicated that a reduced number of patients with severe asthma were seen in 1993 in both Pediatric and Internal Medicine Departments compared with 5 years before, despite the increase in total number of asthma patients in Japan. Specifiic IgE radioallergosorbent test (RAST) measurements were frequently performed instead of skin testing for diagnosis, and eosinophil count and bronchodilator response served as an adjunct to the diagnosis. Patients were frequently asked detailed questions about aspirin-induced asthma, which accounted for 8.8, 2.2 and 1.5% of patients with asthma in the adult, schoolchildren (6–16 years) and infant (≤ 5 years) groups, respectively. In achieving ‘control of asthma’, first priority was given to coping with the symptoms in children aged 5 years or less and to enabling routine daily life activities in patients 6 years of age or older. Usefulness of peak flow measurements was widely recognized and a detailed plan for allergen avoidance (house dust) was often given to patients.

In order to clarify the clinical significance of serum interleukin 4 (IL-4) levels, we measured serum IL-4 concentrations in allergic and non-allergic children using a highly sensitive sandwich ELISA. The limit of detection of the assay was 0.15 pg/ml in serum samples. Serum IL-4 was detected in 96.3% (53/55) of non-allergic controls, in 92.9% (183/197) of allergic children, in 70% (7/10) of cord blood samples and in 86.7% (26/30) of neonates. The IL-4 levels in sera from non-allergic controls were relatively constant during the ages examined and all samples were under 1.5 pg/ml. In allergic children, the serum levels of IL-4 were significantly elevated, particularly at age 13-24 months. The serum levels of IL-4 did not differ in children with different clinical manifestations of allergy, such as bronchial asthma, and atopic dermatitis. The serum level of soluble CD23 (sCD23) showed an age-dependent change in allergic and non-allergic children and was significantly higher in allergic than in non-allergic infants aged 7 to 12 months, but not in other age groups. There was no significant correlation among serum levels of IL-4, sCD23 and IgE.It is suggested that the measurement of serum IL-4 and sCD23 is helpful in the examination of allergic patients in infancy and early childhood, but neither the serum level of IL-4 nor sCD23 directly reflects in vivo IgE production.

Murine embryonal carcinoma (EC) P19 cells, a tissue culture model of early embryonic development, failed to produce cytokines, such as interleukin-3 (IL-3), IL-4, granulocytemacrophage colony stimulating factor (GM-CSF) and interferon-beta (IFN-beta) at the mRNA level. Differentiation induced by retinoic acid (RA) released this repression to produce some cytokines. GM-CSF and IFN-beta genes were expressed in response to PMA/A23187, poly(I):poly(C), IL-1 alpha, forskolin, or LPS stimulation in differentiated P19 cells, whereas IL-3 and IL-4 genes were not expressed. To elucidate the mechanism of the GM-CSF gene induction after differentiation, we transfected a series of 5' deletion mutants of the mouse GM-CSF promoter fused to the bacterial CAT gene. The 740-bp fragment of the 5'-flanking region mediated the positive response. Deletion analysis revealed that the 5' boundary region of the DNA element required for activation lies between positions -95 and -84 and the region upstream of position -95 appears inhibitory. These results indicate that the maturation of the transcriptional machinery after differentiation results in the activation of the GM-CSF gene.

As, especially in children, cognitive function might vary from one stimulus to the other, even when the same target stimuli are presented in the oddball paradigm, we applied the single trial analysis adopting discrete Fourier transformation followed by inverse discrete Fourier transformation to the auditory P300 for the first time in children. In 9 children, age 9–13 years, a positive peak was clearly identified in approximately a quarter of the processed single sweep responses to target stimuli (single sweep P300), whereas it was observed less frequently for non-target stimuli. In 2 children of age 6 years, it was difficult to discriminate single sweep P300 from background EEG activities even in the processed data. As for the former group, the detection ratio of P300 in single sweep target responses ranged from 10.0 to 40.0%, and its mean latency and amplitude ranged from 311 to 348 ms and from 31.7 to 41.5 μV, respectively. There was a significant correlation between average data and single sweep data in terms of the P300 latency. However, the amplitude of P300 in the average data bore no relation to the detection ratio, mean amplitude or variability of latency among single sweep P300s. As compared with adult data previously reported, the detection ratio of single sweep P300 was lower and the P300 amplitude was higher in childhood. P300 in children, at least past puberty, can be analyzed by using the single trial method, which is expected to provide more information about the development of human higher brain functions, especially cognitive function.

We evaluated the neuroprotective effect of the nitric oxide synthesis inhibitor, NG-nitro-L-arginine in a neonatal hypoxic-ischemic rat model. Unilateral hypoxic-ischemic injury was produced in the brain of 7-d-old rats using a combination of a common carotid artery ligation and a hypoxic (8% oxygen) exposure for 2.5 h. In our experimental condition, rectal temperatures did not differ between NG-nitro-L-arginine-treated and saline-injected pups. We killed the animals 72 h later and assessed the hypoxic-ischemic brain damage histologically. NG-nitro-L-arginine (2 mg/kg) administered intraperitoneally 1.5 h before hypoxia resulted in 77% reduction of the infarcted hemispheric volume and 87% reduction of the infarcted striatal volume compared to saline injected controls. NG-nitro-L-arginine given 1.5 h before the insult also significantly prevented hypoxic-ischemic damage in the five hip-pocampal structures examined, dentate gyrus, CA4, CA3, CA1, and subiculum. NG-nitro-L-arginine administered immediately after hypoxia did not prevent hypoxic-ischemic brain damage. These results indicate that nitric oxide plays a key role in producing neonatal hypoxic-ischemic brain damage.

In a previous report,we reported that the induction of GM-GSF gene in differentiated P19 cells results from the maturation of the transcriptional machinery. Here, we identified a cis-DNA element which confers the activation of GM-CSF gene in response to PMA/A23 187 stimulation in differentiated state. Analysis of the 5′-flanking region between −113 and −60 revealed two elements responsible for promotion and one for inhibition, and the overall effects led to the activation of GM-CSF gene mainly through the sequence between −95 and −73. Using the oligonucleotide between −94 and −73 as a probe in gel retardation assays, we identified a DNA-binding protein, NF-GM-P19, the binding activity of which was induced after differentiation in response to PMA/A23 187 stimulation. These results indicate that the induction of GM-CSF gene after differentiation results from the maturation of the transcriptional machinery which recognizes the sequence between −95 and −73.

We measured the binding of [3H]3-[(+/-)2-carboxypiperazin-4-yl] propyl-1-phosphonic acid ([3H]CPP), a competitive ligand for N-methyl-D-aspartate (NMDA) receptors, in double mutant spontaneously epileptic rats (SER: zi/zi, tm/tm) and their parent strains, zitter rats and tremor rats, and WTC rats (control rats from tremor rats derived from Kyoto:Wistar rats) before and after the onset of seizures in tremor rats and SER. Significantly lower [3H]CPP binding receptor density (Bmax) was found in the cortex of SER and zitter rats at 12-15 weeks of age than in that of WTC rats and tremor rats, and at 4 weeks of age the Bmax in zitter rats was lower than that in the other strains. The reduction of Bmax in SER at 12-15 weeks of age may reflect a down regulation of NMDA receptors due to repetitive tonic seizures in SER.

Mutant epileptic El mice are thought to have focal epilepsy of hippocampal origin because glucose utilization is increased in the hippocampus (HPC) during seizures in these mice. However, direct electrographic evidence is still lacking for the notion. We recorded electroencephalograms (EEGs) using depth electrodes in El and non-epileptic ddY mice. All the mice were subjected to a conventional seizure-provoking maneuver during EEG recording; each mouse was placed on a mesh floor and observed for 3 min, and then tossed up in the air. When the El mice showed signs of abortive seizures or prodromal symptoms including squeaking, running and myoclonus, sporadic spikes or sharp waves were generated exclusively in the HPC. When generalized convulsions followed these prodromes, the sporadic discharges evolved into a burst of generalized spikes which again predominated in the HPC. We also observed the cerebral cortex, amygdaloid, caudate, centro-median thalamic and ventral postero-lateral thalamic nuclei, all of which were found to be only secondarily involved. These findings provide the first electrical evidence that El mice have a secondarily generalized seizure that has its initiating focus in the HPC.

We compared regional cerebral blood flow assessed by[123I]N-isopropyl-p-iodoamphetamine (IMP) single-photon emission tomography (SPECT) with magnetic resonance imaging (MRI), computed tomography (CT) and interictal surface electroencephalography (EEG) to evaluate its diagnostic potential in 24 patients with partial seizures with onset in childhood. Focal low uptake areas were observed on SPECT scans of 18 patients and were presumed to represent epileptogenic areas in 17. MRI revealed an abnormality in 12 and CT in 6 patients, and all organic lesions showed SPECT abnormalities, too. Six patients without focal structural abnormalities showed regional perfusion deficit on SPECT. Routine scalp EEG revealed an epileptic focus in 17 patients and three of them showed discordant results between SPECT and EEG, which suggested more serious brain disorders. In two patients without EEG localization only SPECT showed focal abnormalities in the probable epileptic area. [123I]IMP-SPECT was useful in locating the epileptic focus, particularly during the early period after the onset of partial seizures when the EEG gave inconclusive results.

SUMMARY 1. The effects of adenosine and its analogues on actin polymerization in human polymorphonuclear leucocytes (PMN) induced by three different chemotactic stimulants, platelet-activating factor (PAF), N-formyl-methionyl-leucyl-phenylalanine (FMLP) and an activated fragment of C5 (C5a) were investigated. 2. Adenosine and its analogues inhibited the actin polymerization induced by these three agents in a concentration-dependent manner and theophylline, a competitive antagonist at adenosine receptors, abolished these inhibitory effects. 3. The adenosine analogue 5′-N-ethylcarboxamideadenosine (NECA) was a more potent inhibitor of actin polymerization than either l-N6-phenylisopropyladenosine (PIA) or adenosine itself; the rank order of potency of these agonists was characteristic of adenosine A2 receptors. 4. Adenosine deaminase (ADA) abolished the inhibitory effect of adenosine and augmented PAF-induced actin polymerization. 5. It was concluded that, at physiological concentrations, adenosine inhibits actin polymerization in PMN via activation of PMN surface membrane adenosine A2 receptors and thus modulates chemotactic stimulus-induced PMN motility.

Noradrenaline (NA) and dopamine (DA) levels in six brain regions of stimulated and nonstimulated El (El[s] and El[ns]) mice and their maternal ddY mice were determined at various ages and various times after a convulsion. The NA levels in the striatum and hippocampus of 12-week-old El[s] and El[ns] mice were lower than in ddY mice, and remained lower in 23-week-old El[s] mice, but not in El[ns] mice. DA levels were lower in the striatum of El[s] mice than in El[ns] and ddY mice at 16 and 23 weeks of age. NA levels decreased during seizure in the striatum and hippocampus of El[s] mice, and returned to preconvulsive levels 1 hr after convulsion in the striatum and 30 min in the hippocampus. DA levels in the striatum of El[s] mice decreased during convulsion and increased from 1 to 10 min after convulsion. These changes suggest that the NAergic systems in the striatum and hippocampus and the DAergic system in the striatum have important roles in relation to seizure susceptibility in El mice.

The inhibitory effects of azelastine hydrochloride on PAF-induced and fMLP-induced Ca2+ influx, actin polymerization and calcium ionophore A23187-induced and aggregated IgG-induced release of eosinophil cationic protein (ECP) of an eosinophilic leukaemia cell line, EoL-1, were examined. EoL-1 cells cultured with 0.2 mM dibutyryladenosine-cyclic monophosphate for 48 hours showed an increase in intracellular free Ca2+ concentration ([Ca2+]i) and actin polymerization when stimulated by PAF and fMLP. Azelastine hydrochloride inhibited PAF-induced and fMLP-induced Ca2+ influx ([Ca2+]i) in a dose-dependent manner with an IC50 of 1 x 10(-8) M and 1 x 10(-7) M, respectively. It also inhibited PAF-induced and fMLP-induced actin polymerization in a dose-dependent manner up to 40% and 30%, respectively. EoL-1 cells were differentiated to contain ECP in their eosinophilic granules when cultured for 9 days with supernatants of a human adult T cell leukaemia cell line, HIL-3 (HIL-3 sup). Calcium ionophore A23187 and aggregated IgG induced the secretion of ECP by EoL-1 cells. Azelastine hydrochloride inhibited the secretion of ECP in a dose-dependent manner. These inhibitory effects were seen even at therapeutic concentrations of 10(-8) M to 10(-9) M. These results indicate that the therapeutic effects of azelastine hydrochloride as an anti-allergic agent may include inhibition of the accumulation of eosinophils into the locus of allergic inflammation and of the release of cytotoxic granules from eosinophils.

Background. A prospective study was conducted to test the feasibility of nuclear magnetic resonance (NMR) imaging in the early diagnosis of treatment-induced leukoencephalopathy. Methods. The study group included 16 patients with acute lymphoblastic leukemia and 4 patients with malignant lymphoma. All were given intravenous and intrathecal (IT) methotrexate (MTX) for central nervous system prophylaxis. Serial NMR studies were performed before and/or during induction-consolidation cycles. Results. NMR imaging disclosed leukoencephalopathy in 8 of the 20 patients (40%) in the early stages of treatment. In six of the eight, the leukoencephalopathy was resolved after temporary or permanent interruption of IT MTX, and chemotherapy was completed successfully. The other two patients are being treated. Transient neurologic abnormalities developed in two of the eight patients. Conclusions. The possible causal relationship between leukoencephalopathy and the antimetabolic effects of MTX is discussed. This study clearly shows that NMR imaging is valuable in the early diagnosis and management of treatment-induced leukoencephalopathy. Cancer 1992; 70: 1997-2004.

We studied the effect of vasoactive intestinal peptide (VIP), somatostatin (SOM), and substance P (SP) on IL-4-stimulated human IgE and IgG subclass production. VIP and SOM, but not SP, inhibited IgE production without affecting IgM or IgA production by mononuclear cells (MNC) from nonatopic donors from 10 pM to 10 n M . These neuropeptides also differentially modulated IgG subclass production. While IgG1 production was not affected by VIP, SOM, or SP, all of the neuropeptides enhanced IgG2 production. By contrast, SOM and SP, but not VIP, inhibited IgG3 production, whereas VIP and SP, but not SOM, enhanced IgG4 production. The effect by neuropeptides was specific since each peptide effect was specifically blocked by each antagonist. To achieve this effect, neuropeptides must be added at the start of the culture and be present throughout the entire culture period. The inhibition of IgE production was not mediated by known inhibitors of IgE production, IFN-γ or PGE 2 , because the addition of anti-IFN-γ mAb (10 μg/ml) or indomethacin (0.1 μ M ) did not overcome the inhibition of IgE production. In contrast to MNC, neuropeptides did not affect IgG subclass production in purified B cells. IgE production was not induced by IL-4 in purified B cells. Neuropeptides also failed to modulate IgG subclass production in cultures of B cells with either T cells or monocytes. However, they modulated IgE production and IgG subclass production in B cells in the presence of T cells and monocytes. In purified B cells, IL-4 plus anti-CD40 mAb induced IgE production which was not inhibited by VIP or SOM. However, VIP or SOM, but not SP, inhibited IgE production in B cells cultured with both T cells and monocytes. Finally, the mechanism of modulation of IgE and IgG4 production was dependent on IL-4-induced switching, since neuropeptides modulated IgG4 and IgE production in surface IgG4-negative (sIgG4 − ) and sIgE − B cells, respectively. In contrast, modulation of IgG2 and IgG3 production was not due to switching, since neuropeptides did not affect either IgG2 or IgG3 production in sIgG2 − or sIgG3 − B cells, respectively.

The effect of interleukin 8 (IL-8) on IL-4-induced immunoglobulin E (IgE) production was studied. IL-4 induced IgE and IgG4 production by tonsillar mononuclear cells (MNC) without affecting IgM, IgG1, IgA, IgG2, or IgG3 production. IL-8 inhibited IL-4-induced IgE and IgG4 production, whereas it had no effect on IgM, IgG1, IgA, IgG2, and IgG3 production. The inhibitory effect by IL-8 was specific, since it was blocked by anti-IL-8 mAb, but not by control IgG1. Although interferon gamma (IFN-gamma) also inhibited IgE and IgG4 production by MNC stimulated with IL-4, the inhibitory effect of IL-8 was not mediated by IFN-gamma, since the IL-8-induced inhibition could not be blocked by anti-IFN-gamma. Furthermore, anti-IL-8 mAb had no effect on IFN-gamma-induced inhibition. Moreover, addition of IL-5 or IL-6 did not reverse IL-8-induced inhibition of IgE production. In contrast to these observations with MNC, IL-4 failed to induce IgE and IgG4 production by purified B cells. However, combined treatment of purified B cells cells with IL-4 and anti-CD40 antibody resulted in IgE but not IgG4 production. IL-8 inhibited this IgE production without affecting IgM, IgG1, IgG2, IgG3, IgG4, or IgA production, whereas IFN-gamma, IFN-alpha, or prostaglandin E2 (PGE2) failed to do so. These results indicate that IL-8 antagonizes IL-4-induced IgE production by directly affecting B cells through a specific mechanism that is different from IFN-gamma, IFN-alpha, or PGE2.

The effects of several cytokines and phorbol myristate acetate (PMA) on LFA-1 and ICAM-1 expression on a human eosinophilic leukemia cell line, EoL-3, were investigated and compared with those of a human monocytic leukemia cell line, U937. EoL-3 cells expressed large amounts of LFA-1 and small amounts of ICAM-1, and their expression was regulated similarly in EoL-3 cells and U937 cells. Interferon-gamma (IFN-gamma) enhanced ICAM-1 expression but not LFA-1 expression, and PMA augmented both LFA-1 and ICAM-1 expression. IFN-gamma and PMA showed an additive effect on ICAM-1 expression. These results collectively suggest that expression of LFA-1 and ICAM-1 is regulated differently and that IFN-gamma and PMA regulate the expression through different mechanisms. PMA but not IFN-gamma induced homotypic adhesion of EoL-3 and U937 cells, suggesting that PMA but not IFN-gamma activated the adhesive function of these cells. Staurosporin, an inhibitor of protein kinases (PKs), partly suppressed IFN-gamma- and PMA-augmented expression of ICAM-1 on EoL-3 and U937 cells, but did not affect PMA-augmented LFA-1 expression, suggesting that staurosporin-sensitive PKs are involved in IFN-gamma- and PMA-augmented ICAM-1 expression but not in PMA-augmented LFA-1 expression. The role of protein kinase C (PK-C) in these mechanisms was not revealed because a PK-C inhibitor, H-7, did not show any definitive effect on IFN-gamma- and PMA-induced expression of LFA-1 and ICAM-1. Moreover, cyclic AMP (cAMP)- and cGMP-dependent pathways were not shown to be involved in the augmentation of the expression of these molecules.

Adenosine and its analogues inhibited increases in divalent cation influx stimulated by platelet-activating factor (PAF) and formyl-methionyl-leucyl-phenylalanine (FMLP) in a dose-dependent fashion. This effect was antagonized by theophylline, an adenosine receptor antagonist. When extracellular adenosine was removed by adenosine deaminase, the effect of adenosine was completely abolished. Two adenosine analogues with different affinities for adenosine receptor subtypes, 5'-N-ethylcarboxamideadenosine (NECA) and L-N6-phenylisopropyladenosine (PIA), also inhibited divalent cation influx, NECA being more potent than PIA. These results suggest that adenosine and its analogues inhibit divalent cation influx across neutrophil plasma membranes via surface adenosine A2 receptors. Adenosine had little effect on the initial peaks of intracellular free calcium rises induced by chemoattractants, but it inhibited the subsequent rise in free calcium. Since calcium influx through the divalent cation channels or neutrophil plasma membranes is responsible for maintaining free calcium concentration following the initial peaks, we suggest that adenosine modulates neutrophil function by interfering with this calcium influx.

A two-year-old girl was found to have a ring 14 chromosome: [46, XX, r( 14) (P13 q32.3)]. Her development, including verbal ability, was retarded, her CT scan displayed a low density area anterior to the left temporal lobe, and she suffered from complex partial seizures. Focal central nervous system abnormalities may be present in patients with ring 14 chromosome, and their seizures are not exclusively of the parimary generalized type. This is the first case with ring 14 chromosome and complex partial seizures. Key words: Ring 14 chromosome, epilepsy, complex partial seizure, focal CNS disturbance.

Tremor rats begin to exhibit clinical or electrical absence-like seizures after 6 weeks of age, and by 14 weeks of age, all have seizures. Central-type benzodiazepine receptor binding was investigated in tremor rats and control rats, aged 4 weeks and 16 weeks. Significantly lower benzodiazepine receptor density and no differences in affinity were found in the hippocampus of the tremor rats in comparison with that of control rats at both ages. This abnormality is considered to be due to a tremor gene and may be the cause of absence-like seizures in tremor rats. A significantly lower receptor density was found in the cerebellum at 4 weeks of age in the tremor rats than in the control rats. These changes may be related to tremorous movements in the tremor rats. Receptor density was significantly lower in the brainstems of tremor rats and control rats at 16 weeks of age than at 4 weeks of age, and the decrease was more marked in control rats. These facts may reflect a reduced decrease in the response to the dysfunction of gamma-aminobutyric acidergic neurons, or the function of the gamma-aminobutyric acid/benzodiazepine receptor system may be secondarily increased to suppress seizures in 16-week-old tremor rats.

The effect of eosinophil cationic protein (ECP) on immunoglobulin (Ig) production by and proliferation of human plasma cells was studied. ECP inhibited Ig production by and proliferation of the human plasma cell lines, IM-9 and AF-10, in a dose-dependent fashion. As little as 0.05 ng/ml ECP was found to be inhibitory, and the maximal inhibition was achieved at doses of 0.1-0.5 ng/ml ECP. This inhibition was not due to cytotoxicity, since viability was always greater than 98%. Kinetic experiments demonstrated that inhibition was observable after 24 hr of culture with ECP and that the inhibitory effect of ECP was reversible. The inhibitory effect of ECP could be blocked by anti-ECP serum, but not by control serum. Of the various cytokines tested, including interleukin (IL)-1β, IL-2, IL-3, IL-4, IL-5, IL-6, interferon (IFN)-α, IFN-γ, granulocyte-macrophage colony-stimulating factor (GM-CSF) and erythropoietin (Epo), IL-6 reversed the inhibition, while other cytokines failed to do so. ECP also inhibited Ig (IgG1, IgG2, IgG3, IgG4, IgM, and IgA) production by and proliferation of PCA-1 + plasma cells generated in vitro with a similar doseresponse pattern. This inhibition also was blocked by anti-ECP serum but not by control serum, and was restored by IL-6. These results suggest that ECP may interact with IL-6 in controlling plasma cell responses.

SUMMARY The effect of vasoactive intestinal peptide (VIP) on human lymphoblastoid B cell lines and tonsil B cells was studied. VIP increased immunoglobulin production and proliferation by lymphoblastoid B cell line. GM-1056, in a dose-dependent manner. As little as 10-12 M of VIP was effective, and higher concentrations of VIP induced an approximately five-fold increase in IgA production. Moreover, this enhancement was blocked by VIP antagonist. Similarly, VIP enhanced IgM and IgG production by other lymphoblastoid B cell lines, CBL and IM-9, respectively. In contrast to VIP, another neuropeptide substance P (SP) or somatostatin failed to enhance immunoglobulin production and thymidine uptake. VIP also enhanced IgA production and thymidine uptake by purified tonsil B cells. However, in contrast to B cell lines, VIP failed to enhance IgM and IgG production by tonsil B cells. SP or somatostatin failed to enhance immunoglobulin production or thymidine uptake by tonsil B cells. These results indicate that VIP acts as B cell stimulatory factor and that VIP may also have preferential effect on IgA production on tonsil B cells.

Superoxide production by human neutrophils stimulated with FMLP and soluble aggregated human IgG were inhibited in a dose dependent manner by two kinds of tyrosine kinase inhibitors, erbstatin and genistein. Superoxide production stimulated with surface bound IgG, however, was scaroely inhibited by either inhibitor. Protein tyrosine phosphorylation studies with immunoblotting revealed specific tyrosine phosphorylation of a 40 Kd protein by soluble aggregated and surface bound IgG, and that of a 39 Kd protein, as well as the 40 Kd protein, by FMLP. These were all inhibited by the tyrosine kinase inhibitors. These data suggest that superoxide production induced by FMLP and soluble aggregated IgG are, at least in part, tyrosine kinase dependent, but the tyrosine kinases and/or substrates of tyrosine kinases involved may be different. In addition, tyrosine kinase independent pathways are also suggested to be involved in superoxide production by stimulation with surface bound IgG.

We correlated serum IgE, interleukin 4 (IL4) and soluble low affinity Fc receptor for IgE (sFcɛRII) in atopic and nonatopic children. Serum sFcɛRII was significantly higher in atopic than in nonatopic children (0-2 yr). Serum sFcɛRII correlated closely with serum IgE in infants, but not in older children. To study further the characteristics of IgE synthesis in infants, we measured the serum level of IL4, which has been demonstrated to enhance the production of both IgE and sFcɛRIIin vitro. Serum IL4 was significantly higher in atopic than in nonatopic infants, but there was no significant difference between atopic and nonatopic older children. These results suggest that IL4 plays an important role in the regulation of IgE synthesis in infants and that IgE synthesis in older children might be modulated by other factors.

We investigated tyrosine phosphorylation and structural properties of the IgM-associated molecules in comparison with IgD-associated molecules in a recently established human surface IgM+, IgD+ B lymphoma cell line, B104, the growth of which was irreversibly inhibited by anti-IgM mAbs but not by anti-IgD mAbs. Tyrosine kinase activity and tyrosine phosphorylated proteins were detected in anti-IgM and anti-IgD immunoprecipitates from digitonin lysates of B104 cells with the use of an in vitro kinase assay followed by a re-immunoprecipitation experiment with anti-phosphotyrosine mAbs. Tyrosine phosphorylated proteins of 74, 58-44, 41, and 39 kDa were detected in anti-IgM immunoprecipitates, whereas tyrosine phosphorylated proteins of 74, 58-44, and 39 kDa, but not 41 kDa, were detected in anti-IgD immunoprecipitates. Crosslinking of surface IgM and surface IgD stimulated rapid tyrosine phosphorylation of different sets of proteins which included tyrosine-phosphorylated proteins of the same or similar molecular weights as those detected in the anti-IgM and anti-IgD immunoprecipitates respectively. After deglycosylation by N-glycosidase, both the IgM- and IgD-associated phosphoproteins (pp58-pp39) gave rise to the same three bands of 29, 27, and 26 kDa. Proteolytic peptide mapping of these three deglycosylated proteins showed that the primary structures of the IgM- and IgD-associated molecules are identical, suggesting that the IgM- and IgD-associated phosphoproteins (pp58-pp39) are the products of the same or closely related genes. One of the products, pp41, may be associated with IgM, but not with IgD, although the same gene product may be associated with IgD in a different glycosylation pattern.

To identify the focus of paroxysmal neuronal activity causing epilepsy in tuberous sclerosis (TS), the regional cerebral blood flow (r-CBF) in 19 patients with TS was assessed using single-photon emission computed tomography (SPECT) with I-123 iodoamphetamine (IMP), in correlation with serial interictal EEGs and organic changes observed on magnetic resonance imaging (MRI). There was a general irregularity of cortical IMP uptake and retention in TS, and two-thirds of the cerebral regions exhibiting high intensity in T2-weighted MRI images (cortical tubers) showed a decrease in r-CBF. In addition to in tubers, decreased r-CBF was observed in regions in which MRI was considered to indicate destruction of the normal cortex, atrophy or vascular abnormalities, although these areas did not consistently show epileptic changes in serial EEGs. Among the cortical regions which consistently showed epileptic foci in serial EEGs, none showed abnormal r-CBF without lesions on MRI. We conclude that IMP-SPECT is useful for visualizing the epileptogenic laterality in cases with bilateral MRI lesions and EEG epileptic changes, and to differentiate epileptogenic foci from electrophysiological propagated areas. However, regarding the severity of epilepsy, the MRI findings showed a better correlation than the IMP-SPECT findings did.

We investigated the somatosensory evoked potentials (SEPs) produced by posterior tibial nerve (PTN) stimulation in 8 infants and children with focal spinal cord disorders. The spinal responses of the PTN-SEPs were considered to assist in the localization of spinal lesions because their abnormalities were consistent with the neurologic and/or radiologic findings in all 6 examinations that revealed abnormal spinal SEPs. The cortical responses correlated significantly with proprioception in the lower limbs because proprioception was only disturbed when the cortical SEPs were absent (3 examinations). When both the spinal and cortical responses were abnormal, the spinal lesion probably involved the dorsal column so extensively that it completely interrupted the afferent impulses. In contrast, when SEP studies demonstrated abnormal spinal and normal cortical SEPs (3 examinations), the dorsal column involvement was probably less severe; therefore, both the spinal and cortical responses provided useful information regarding afferent conduction in the dorsal column. PTN-SEPs appear to have the potential to be of value in the diagnosis of focal spinal disease, especially in infants and young children who cannot cooperate with detailed neurologic examinations.

Differentiation of a human eosinophilic leukemia cell line, EoL-1, induced by the culture supernatant of a human ATL cell line, HIL-3 (HIL- 3 sup) was compared with differentiation induced by defined cytokines. HIL-3 sup induced EoL-1 cells to express eosinophilic granules and segmented nuclei after 6 to 9 days of incubation. HIL-3 sup also induced the expression of Fc epsilon receptor II (Fc epsilon RII/CD23) and an eosinophil differentiation antigen EO-1 mainly on eosinophilic granule (+) cells. Furthermore, HIL-3 sup induced EoL-1 cells to respond to an eosinophil chemotactic factor, platelet activating factor. HIL-3 cells express messenger RNA (mRNA) of interleukin-5 (IL- 5), macrophage colony-stimulating factor (M-CSF), and IL-3 but not granulocyte CSF (G-CSF). Granulocyte-macrophage CSF (GM-CSF) and tumor necrosis factor-alpha (TNF-alpha) were detected in the HIL-3 sup. Recombinant IL-2 (rIL-2), rIL-3, rIL-4, rIL-5, rM-CSF, and rGM-CSF did not induce eosinophilic granules. rG-CSF induced a few eosinophilic granule (+) cells, and TNF-alpha, which did not induce eosinophilic granules by itself, enhanced the ability of G-CSF to induce them. However, G-CSF and TNF-alpha did not induce the expression of Fc epsilon RII and EO-1 antigen. Moreover, anti-G-CSF, anti-TNF-alpha, anti-GM-CSF, anti-IL-3, and anti-IL-5 antibodies did not suppress the effect of HIL-3 sup on the differentiation of EoL-1 cells. All the data suggest that HIL-3 sup contains an unidentified factor that induces differentiation of EoL-1 cells, and that EoL-1 cells and HIL-3 sup provide an important model for the examination of differentiation mechanisms and functions of eosinophils.