Mucor pulmonary embolism in a patient with myelodysplastic syndrome.

Mucormycosis is a life-threatening infectious disease that occurs most commonly in immunocompromised patients such as those with hematological malignancies. Its clinical symptoms and associated radiological findings vary and specific biomarkers and culture characteristics have not been defined. An 85-year-old man who had been treated for myelodysplastic syndrome and tuberculosis for several months presented with subacute fever and worsening left-side chest pain. Contrast-enhanced computed tomography images depicted massive tumor-like consolidation without enhancement, expanding from the left lower lobe. Emboli that did not respond to anticoagulants were detected in the left descending pulmonary artery. Despite intensive treatment he developed multiple organ failure and died 47 days after hospitalization. Gross pathology of a lung autopsy specimen revealed left lower pulmonary arterial emboli and pulmonary infarction, which was concluded to be the direct cause of death. The emboli were histopathologically identified as invasive mycelia in vessels. Mucor sp. was detected via real-time polymerase chain reaction and immunohistopathological analyses revealed that the mold in the blood vessels of lung tissue was partially positive for the mucor antigen. In the present case of Mucor sp. pulmonary emboli in a patient with myelodysplastic syndrome, radiographic findings were hard to distinguish from those typical of a lung abscess.

Cost-effectiveness of interferon-gamma release assay for systematic tuberculosis screening of healthcare workers in low-incidence countries.

BACKGROUND: Tuberculosis (TB) is one of the important occupationally acquired infectious diseases in low-incidence countries. Delays in TB diagnosis and treatment among healthcare workers (HCWs) result in costly large-scale TB contact screening among patients and other HCWs. AIM: To assess the cost-effectiveness of TB screening for HCWs using interferon-gamma release assays (IGRAs) compared with tuberculin skin test (TST) and chest x ray (CXR). METHODS: Markov models were constructed using a hospital payer perspective. The target populations were a hypothetical cohort of 30-year-old HCWs at the time of employment, and a hypothetical cohort of HCWs working on a high-risk ward until 60 years of age. Six strategies were modelled: TST, QuantiFERON-TB Gold In-Tube (QFT), T-SPOT.TB (T-SPOT), TST followed by QFT, TST followed by T-SPOT, and CXR. The main outcome measure of effectiveness was quality-adjusted life-years (QALYs). Costs and QALYs gained per person screened were calculated. FINDINGS: QFT was the most cost-effective strategy at the 'willingness to pay' level of US$ 50,000/QALYs gained (at the time of employment: US$ 334.91, 21.071 QALYs; on a high-risk ward: US$ 1050.32, 20.968 QALYs; values for 2012). Cost-effectiveness was sensitive to latent TB infection (LTBI) rate and bacillus Calmette-Guérin vaccination rate. TST followed by QFT was more cost-effective than QFT when the LTBI rate was <0.026 at the time of employment and <0.08 on a high-risk ward. CONCLUSION: Systematic TB screening using QFT is cost-effective for screening HCWs, and is recommended in low-incidence countries.

Genetic diversity of Mycobacterium tuberculosis isolates from foreign-born and Japan-born residents in Tokyo.

Sequences of the full genomes of 259 clinical isolates of Mycobacterium tuberculosis, obtained from foreign-born and Japan-born patients in Tokyo, Japan, were determined, and a phylogenetic tree constructed by concatenated single-nucleotide polymorphism (SNP) sequences. The 259 isolates were clustered into four clades: Lineage 2 (East Asian or "Beijing" genotype; n = 182, 70.3%), Lineage 4 (Euro-American, n = 46, 17.8%), Lineage 1 (Indo-Oceanic, n = 23, 8.9%), and Lineage 3 (East African-Indian, n = 8, 3.1%). Of the 259, 36 (13.9%) were resistant to at least one drug. There was no multi-drug-resistant isolate. Drug resistance was greater for the strains in Lineage 2 than the non-Lineage 2. The proportion of Lineage 2 isolates was significantly smaller in foreign-born (n = 43/91, 47.3%) than in Japan-born (n = 139/168, 82.7%) patients, whereas the proportion of Lineage 1 isolates was significantly larger in foreign-born (n = 19/91, 20.9%) than in Japan-born (n = 4/168, 2.4%) patients. We also found eight SNPs specific to the typical Beijing sub-genotype in Lineage 2, including 4 non-synonymous SNPs. Of the 259 isolates, 244 had strain-specific SNP(s) and small (1-30-bp) insertions and deletions (indels). The numbers of strain-specific SNPs and indels per isolate were significantly larger from foreign-born (median 89, range 0-520) than from Japan-born (median 23, range 0-415) (p 3.66E-15) patients. These results suggested that M. tuberculosis isolates from foreign-born patients had more genetic diversity than those from Japan-born patients.

Leukemoid reaction and chronic lung disease in infants with very low birth weight.

OBJECTIVES: To analyze the relationship between the leukemoid reaction and chronic lung disease in very-low-birth-weight (VLBW) infants. METHODS: Neonates born weighing less than 1500 g without evidence of congenital anomalies and admitted to our hospital from October 1985 to December 1999 comprised our study. Leukemoid reaction was defined as a peripheral white blood cell (WBC) count of > or = 50 x 10(3)/microl. The infants who demonstrated a leukemoid reaction formed the study group, while the remainder formed the control group. The relationship between neonatal variables and WBC counts was studied. RESULTS: Fourteen of the 486 infants demonstrated WBC counts of > or = 50 x 10(3)/microl, with an incidence of 2.9%. Univariate analysis demonstrated a significant association between a leukemoid reaction and chronic lung disease following intrauterine infection. CONCLUSION: A leukemoid reaction was observed in 2.9% of VLBW infants in our neonatal intensive care unit. A significant association was demonstrated between the leukemoid reaction and chronic lung disease following intrauterine infection.

Functional characterization of cysteinyl leukotriene CysLT(2) receptor on human coronary artery smooth muscle cells.

Cysteinyl leukotrienes (LTC(4), LTD(4), and LTE(4)) are a class of biologically active lipids that exert potent effects on the heart. To assess their roles, we investigated the distribution of their receptors, CysLT(1) and CysLT(2), in the cardiovascular system. CysLT(2) mRNA was detected at high levels in the human atrium and ventricle and at intermediate levels in the coronary artery, whereas CysLT(1) mRNA was barely detected. Further analysis by in situ hybridization revealed that CysLT(2) mRNA was expressed in myocytes, fibroblasts, and vascular smooth muscle cells, but not in endothelial cells. When human coronary smooth muscle cells were stimulated with LTC(4), the intracellular calcium concentration increased in a dose-dependent manner, and this action was partially inhibited by nicardipine. Additionally, these cells showed chemotactic responses to LTC(4). This is the first report on the physiological role of CysLT(2), and the findings suggest that CysLT(2) has biological significance in the cardiovascular system.

Molecular cloning of the platelet P2T(AC) ADP receptor: pharmacological comparison with another ADP receptor, the P2Y(1) receptor.

Platelet activation plays an essential role in thrombosis. ADP-induced platelet aggregation is mediated by two distinct G protein-coupled ADP receptors, Gq-linked P2Y(1), and Gi-linked P2T(AC), which has not been cloned. The cDNA encoding a novel G protein-coupled receptor, termed HORK3, was isolated. The HORK3 gene and P2Y(1) gene were mapped to chromosome 3q21-q25. HORK3, when transfected in the rat glioma cell subline (C6-15), responded to 2-methylthio-ADP (2MeSADP) (EC(50) = 0.08 nM) and ADP (EC(50) = 42 nM) with inhibition of forskolin-stimulated cAMP accumulation. 2MeSADP (EC(50) = 1.3 nM) and ADP (EC(50) = 18 nM) also induced intracellular calcium mobilization in P2Y(1)-expressing cells. These results show that HORK3 is a Gi/o-coupled receptor and that its natural ligand is ADP. AR-C69931 MX and 2MeSAMP, P2T(AC) antagonists, selectively inhibited 2MeSADP-induced adenylyl cyclase inhibition in HORK3-expressing cells. On the other hand, A3P5PS, a P2Y(1) antagonist, blocked only 2MeSADP-induced calcium mobilization in P2Y(1)-expressing cells. HORK3 mRNA was detected in human platelets and the expression level of HORK3 was equivalent to that of P2Y(1). These observations indicate that HORK3 has the characteristics of the proposed P2T(AC) receptor. We have also determined that [(3)H]2MeSADP binds to cloned HORK3 and P2Y(1). Competition binding experiments revealed a similarity in the rank orders of potency of agonists and the selectivity of antagonists as obtained in the functional assay. These results support the view that P2Y(1) functions as a high-affinity ADP receptor and P2T(AC) as a low-affinity ADP receptor in platelets.

Pivotal role of 5-lipoxygenase in the activation of human eosinophils: platelet-activating factor and interleukin-5 induce CD69 on eosinophils through the 5-lipoxygenase pathway.

CD69 is an activation-related cell surface molecule on human eosinophils. It has been reported that interleukin (IL)-5, but not platelet-activating factor (PAF), can induce CD69 on human eosinophils in vitro. In this study, PAF induced CD69 intensely on eosinophils from patients with hypereosinophilic syndrome (HES), while only weakly on those from normal donors. Because HES eosinophils contain abundant cytosolic phospholipase A2 (cPLA2) and 5-lipoxygenase (5-LO), we examined the roles of several enzymes involved in the metabolism of arachidonic acid in the PAF- or IL-5-induced CD69 expression on eosinophils. The CD69 expression induced by PAF and IL-5 on HES eosinophils and that by IL-5 on normal eosinophils were both inhibited by AA861 and MK-886, inhibitors of 5-LO activity. In addition, AACOCF3, a selective cPLA2 inhibitor, inhibited IL-5-induced CD69 expression on normal eosinophils, although it hardly affected either IL-5- or PAF-induced CD69 expression on HES eosinophils. Moreover, PAF alone induced CD69 only weakly on normal eosinophils, but exogenous arachidonic acid remarkably enhanced PAF-induced CD69 expression on them. These findings suggest that IL-5 activates both cPLA2 and 5-LO but PAF activates only 5-LO. It is suggested that 5-LO plays a critical role in the induction of CD69 on eosinophils.

Anti-interleukin-8 autoantibody in the tracheobronchial aspirate of infants with chronic lung disease.

BACKGROUND: A high concentration of interleukin (IL)-8 has been observed in the tracheobronchial aspirate of infants with chronic lung disease (CLD), although the pattern varies depending on the type of CLD. Alveolar fluid from patients with adult respiratory distress syndrome (ARDS) also contains an elevated level of IL-8. Recently, the presence of anti-IL-8 autoantibody was demonstrated in the alveolar fluid from patients with ARDS. METHODS AND RESULTS: The concentration of anti-IL-8 autoantibody in the tracheobronchial aspirate of infants with CLD was measured in order to discover whether there was any correlation with the concentration of IL-8. Similar to IL-8 concentration, the anti-IL-8 IgM antibody concentration in all infants with CLD following intrauterine infection was already high during the first 48 h. However, the concentration in infants with CLD following respiratory distress syndrome began to increase after 11 days of life, in contrast with the rise in IL-8 between 48 h after birth and day 5. CONCLUSIONS: The presence of anti-IL-8 autoantibody may provide a mechanism that limits the bioavailability of free IL-8 in the lungs. In addition, the time lag between the increase in IL-8 and anti-IL-8 IgM autoantibody demonstrated in the present study could be used to estimate the time when the inflammation begins, even if the IL-8 concentration is already high.

[Comparison of five guidelines of community-acquired pneumonia].

The aim of this retrospective study was to elucidate the characteristics of five guidelines of community-acquired pneumonia: ATS (1993), ATS (2001), IDSA (1998), IDSA (2000) and the guidelines of the Japan Respiratory Society (2000). One hundred community-acquired pneumonia patients admitted to the International Medical Center of Japan were investigated in accordance with each set of guidelines based on the physical, laboratory, and chest radiography findings on the first day of treatment. According to the ATS (1993) guidelines, 33% of the cases were classified as "severe" pneumonia. On the other hand, according to the ATS (2001) guidelines, only 8% of the cases were classified as "severe" pneumonia. According to the IDSA guidelines, 35% of the patients were classified as "outpatients". Fluoroquinolone appears to be a very important antibiotic drug in the new guidelines of both ATS and IDSA. The scoring system of IDSA suggested a correlation between the patient's score and the pathogenic bacteria. According to the guidelines of the Japan Respiratory Society, 42% of the cases were classified as "severe" pneumonia. There are evident differences between these guidelines, and clinicians need to have a full understanding of their respective characteristics.

[Serial measurement of anti-interleukin-8 IgG autoantibody in cerebrospinal fluid of infants with bacterial meningitis].

We serially measured concentrations of interleukin (IL)-8 and anti-IL-8 IgG autoantibody in cerebrospinal fluid of infants with bacterial meningitis, and also measured these concentrations in cerebrospinal fluid obtained from infants without meningitis on admission. We have reported that the IL-8 concentration in cerebrospinal fluid of infants with purulent meningitis rapidly decreases after the initiation of therapy. Thus, in the present study, the IL-8 concentration in infants with purulent meningitis only before the initiation of therapy was significantly higher compared with that in infants without meningitis. However, the concentration of anti-IL-8 IgG autoantibody was still high after the initiation of therapy. The concentration of anti-IL-8 IgG autoantibody was significantly higher compared with that in infants without meningitis until the 15th day after the initiation of therapy. The time lag between the decrease of IL-8 and anti-IL-8 IgG autoantibody demonstrated in the present study could be used to indicate the past presence of a large amount of IL-8, even if the IL-8 concentration was already low.

Molecular cloning and characterization of another leukotriene B4 receptor.

Leukotriene B(4) is a potent lipid mediator known to be implicated mainly in inflammatory actions. Previous pharmacological studies indicated the existence of only one class of G protein-coupled receptor for leukotriene B(4), for which a candidate gene, namely BLT, had been identified. Here we report the isolation of another gene encoding a functional G protein-coupled receptor for leukotriene B(4), named JULF2. JULF2 is a novel G protein-coupled receptor of 358 amino acids that shares 36.6% amino acid identity with human BLT. According to genomic information, the JULF2 gene is located on the chromosome 14, about 4 kilobases upstream of the BLT gene. During screening of endogenous ligands for JULF2, we found that leukotriene B(4) induced inhibition of forskolin-stimulated cAMP accumulation in Chinese hamster ovary cells, stably expressing JULF2. Additionally, Chinese hamster ovary cells expressing exogenous JULF2 showed chemotactic responses with leukotriene B(4) in a pertussis toxin-sensitive manner. A large amount of JULF2 mRNA was detected in the human spleen and the peripheral blood leukocytes. Furthermore, JULF2 mRNA was expressed in mononuclear lymphocytes, in which BLT mRNA was barely detected. The discovery of this second leukotriene B(4) receptor will eventually lead to a better understanding of the classification of leukotriene B(4) receptors and reconsideration of the pathophysiological role of leukotriene B(4).

The molecular characterization and tissue distribution of the human cysteinyl leukotriene CysLT(2) receptor.

Cysteinyl leukotrienes (CysLTs), slow-reacting substances of anaphylaxis, are lipid mediators known to possess potent proinflammatory action. Pharmacological studies using CysLTs indicate that at least two classes of G protein-coupled receptors (GPCRs), named CysLT(1) and CysLT(2), exist; the former is sensitive and the latter is resistant to the CysLT(1) antagonists currently used to treat asthma. Although the CysLT(1) receptor gene has been recently cloned, the molecular identity of the CysLT(2) receptor has remained elusive. Here we show that the pharmacological profile of an orphan GPCR (PSEC0146) is consistent with that of the CysLT(2) receptor. In human embryonic kidney 293 cells that express the PSEC0146 cDNA, leukotriene C(4) (LTC(4)) and leukotriene D(4) (LTD(4)) induce equal increases in intracellular calcium mobilization; these increases are not affected by CysLT(1) antagonists. Additionally, [(3)H]LTC(4) specifically binds to membranes from COS-1 cells transiently transfected with PSEC0146. Large amounts of the PSEC0146 mRNA are found in human heart, placenta, spleen, and peripheral blood leukocytes but not in the lung and the trachea. Pharmacological feature and expression studies will eventually lead to a better understanding of the classification of CysLT receptors, possibly leading to a reconsideration of the pathological and physiological role of CysLTs.

An evolutionarily conserved G-protein coupled receptor family, SREB, expressed in the central nervous system.

We report here a novel family of G-protein coupled receptor (GPCR) which is extraordinarily conserved among vertebrate species. This family, designated SREB (Super Conserved Receptor Expressed in Brain), consists of at least three members, termed SREB1, SREB2, and SREB3. SREB members share 52-63% amino acid identity with each other and show relatively high similarity to previously known amine amine GPCRs (approximately 25% identity). Amino acid sequence identity between human and rat orthologues is 97% for SREB1 and 99% for SREB3, while the SREB2 sequence is surprisingly completely identical between the species. Furthermore, amino acid sequence of zebrafish SREB2 and SREB3 are 94 and 78% identical to mammal orthologues. Northern blot analysis revealed that SREB members are predominantly expressed in the brain regions and genital organs. Radiation hybrid analysis localized SREB1, SREB2, and SREB3 genes to different human chromosomes, namely 3p21-p14, 7q31 and Xp11, respectively. The high sequence conservation and abundant expression in the central nervous system suggest the existence of undiscovered fundamental neuronal systems consisting of SREB family members and their endogenous ligand(s).

Induction of the activation-related antigen CD69 on human eosinophils by type IIA phospholipase A2.

OBJECTIVE: High levels of human type IIA phospholipase A2 (PLA2-IIA) have been found in the eosinophil-mediated inflammation sites, although the pathophysiological role of PLA2-IIA in the eosinophil activation has remained poorly understood. We investigated the effects of PLA2-IIA on eosinophil activation. METHODS: Eosinophils were incubated with recombinant human PLA2-IIA or other stimuli, and then eosinophil peroxidase (EPO) (by colorimetric assay) and leukotriene C4 (by enzyme immunoassay) released in the incubation buffer were measured. Expression of CD11b and CD69 on the cell surface was also measured by flow cytometry (by mean fluorescence intensity (MFI)). EPO, LTC4, and CD11b are thought to be markers for early phase activation (occurred in an hour after stimulation), and CD69 is to be a marker for late phase activation (occurred after several hours). RESULTS: While PLA2-IIA (5 microg/ml) did not induce any early phase activation, it induced significant expression of an activation-related antigen, CD69, on human blood eosinophils. The PLA2-IIA, when enzymatically inactivated by either p-bromophenacyl bromide or EDTA, lost its effect on the CD69 induction. Similarly to PLA2-IIA, several lysophospholipids (1 microg/ml) also induced CD69 on eosinophils significantly (control, 0.71 +/- 0.11; PLA2-IIA, 3.29 +/- 0.37*; lysophosphatidic acid, 2.57 +/- 0.43*; specific MFI +/- S.E.M., n = 4, * indicate p < 0.05 vs. control). CONCLUSIONS: PLA2-IIA induces CD69 expression on the eosinophils through its catalytic activity at least partly via the enzymatic products such as several lysophospholipids from the eosinophil membrane phospholipids. PLA2-IIA may contribute to the eosinophilic inflammation synergistically with other factors.

Anti-interleukin-8 auto-antibodies in cerebrospinal fluid of children with purulent meningitis.

BACKGROUND: Cerebrospinal fluid (CSF) of patients with purulent meningitis contains a high concentration of interleukin (IL)-8. Recently, the presence of anti-IL-8 auto-antibodies was noted in blood and alveolar fluid. Therefore, measurement of the concentration of anti-IL-8 auto-antibodies was attempted in CSF of children with and without meningitis. METHODS AND RESULTS: We measured the concentration of anti-IL-8 auto-antibodies in CSF of children with purulent or aseptic meningitis and those without meningitis. The CSF obtained on admission showed a significantly higher concentration of anti-IL-8 IgG and IgM auto-antibodies in children with purulent meningitis, compared with those with aseptic meningitis or without meningitis. Among the three groups of children, the concentration of IL-8 was also significantly higher in CSF of children with purulent meningitis. CONCLUSION: Because the anti-IL-8 IgG auto-antibody binds to IL-8 and inhibits IL-8 interaction with specific receptors on neutrophils, the presence of anti-IL-8 auto-antibodies seems to provide a mechanism that limits the bioavailability of free IL-8 in CSF.

The novel G-protein coupled receptor SALPR shares sequence similarity with somatostatin and angiotensin receptors.

A cDNA encoding a novel G-protein coupled receptor (GPCR) was isolated from a human cerebral cortex cDNA library by low stringency hybridization screening. This putative seven-transmembrane domain receptor of 469 amino acids was designated SALPR (Somatostatin- and Angiotensin- Like Peptide Receptor). SALPR shares the highest amount of amino acid similarity with the somatostatin (35% with SSTR5) and angiotensin receptors (31% with AT1). Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that the SALPR mRNA is predominantly expressed in human brain regions, particularly the substantia nigra and pituitary, although the mRNA can also be detected in the peripheral tissues, albeit at low levels. Chromosomal mapping by radiation hybrid analysis localized the human SALPR gene to the chromosome 5p15.1-5p14. Transient expression of SALPR in COS-1 cells did not produce any binding sites for somatostatin or angiotensin II, indicating the necessity for further study to discover its ligand and physiological significance.

Interleukin 8 in the tracheobronchial aspirate of infants acts as a neutrophil chemotactic factor in the development of chronic lung disease.

BACKGROUND: We have already reported that there are some periods when a high interleukin 8 (IL-8) concentration is observed in the tracheobronchial aspirate of infants with chronic lung disease (CLD), although the changing pattern of the IL-8 concentration varies depending on the type of CLD. Interleukin 8 is known as a neutrophil chemotactic agent. Therefore, we asked whether IL-8 is an important neutrophil chemotactic factor in the tracheobronchial aspirate of infants who later develop CLD. METHODS AND RESULTS: We measured the neutrophil chemotactic activity of the tracheobronchial aspirate in CLD infants with or without anti-IL-8 antibody. Preincubation with anti-IL-8 immunoglobulinG resulted in a significant reduction of neutrophil chemotactic activity in the tracheobronchial aspirate. In infants with CLD following respiratory distress syndrome, there was a significant relationship between the IL-8 concentration and the neutrophil chemotactic activity of tracheobronchial aspirate without anti-IL-8 antibody, although no significant relationship was seen in infants with CLD following intra-uterine infection or with other CLD. CONCLUSIONS: Interleukin 8 in the tracheobronchial aspirate seems to play a significant role in recruiting neutrophils into the airways of patients with CLD, especially CLD following respiratory distress syndrome. We believe that in this type of CLD, IL-8 in the lung is generated as a result of hyperoxia rather than infection. In this situation, production of other neutrophil chemotactic factors or some factors that inhibit IL-8 activity may be insignificant.

Strategy for the prevention and treatment of chronic lung disease of the premature infant.

Based on the analyses of approximately 1,000 infants with CLD, this condition was classified into 7 types according to the preceding illnesses and the chest X-ray appearance. The profile of inflammatory enzymes, cytokines and chemical mediators supported the relevance of the classification. Since different insults to the lung with different onsets exhibit the different spectra of the disease, appropriate strategies adapted to each type of CLD should be pursued. For types I and II prophylactic administration of exogenous surfactant, early enough to prevent the oxygen toxicity and barotrauma, is important. RDS can be diagnosed by the stable microbubble rating on gastric aspirates within several minutes of birth. The application of milder modes of ventilation such as HFOV started at birth should also be remembered. The recommended strategies for type IV and V are the care of the very low birth weight infant in the fully humidified incubator with careful fluid administration to prevent symptomatic PDA, and early detection and treatment of infection by screening with sensitive methods such as the APR score. However, the most difficult problem is to identify the correct strategy for type III and III', because these types of CLD can only be prevented by the complete eradication of intrauterine infection.

Cytosolic phospholipase A2, increased and activated in the eosinophils of patients with hypereosinophilic syndrome in vivo, is involved in the augmented release of leukotriene C4.

OBJECTIVE: Eosinophils from patients with hypereosinophilic syndrome (HES) showed augmented release of leukotriene C4 (LTC4) by stimulation with A23187. In the present study, we investigated the involvement of cytosolic phospholipase A2 (cPLA2) in this phenomenon. METHODS: Eosinophils from normal and HES donors (2.5 x 10(6) cells/ml) were incubated with A23187 (0.03-3 microM) for 60 min in the presence or absence of a cPLA2 inhibitor, AACOCF3. The LTC4 released from eosinophils was measured by enzyme immunoassay. Distribution of cPLA2 and 5-lipoxygenase (5-LO) proteins within the eosinophils were detected by Western immunoblotting. RESULTS: The level of LTC4 released from the HES eosinophils by stimulation with A23187 was higher than that from normal eosinophils. The A23187-induced LTC4 release was inhibited by AACOCF3 in a dose-dependent manner. The amounts of cPLA2 seemed to be increased in the non-stimulated HES eosinophils by an analysis of immunoblotting. To be noticed was that cPLA2 was detected as a phosphorylated and membrane-bound form in the HES eosinophils, but not in the normal eosinophils. In contrast, localization of 5-LO within the eosinophils under A23187 stimulation was not different between normal and HES donors, while the amounts of 5-LO also seemed to be increased in the HES eosinophils. CONCLUSIONS: These results suggest that cPLA2, increased and activated (phosphorylated and membrane-translocated) in vivo, is involved in the augmented release of LTC4 from the HES eosinophils.