bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of Multivariate Tests for Genetic Linkage
Authors: Amos Christopher I. , de Andrade Mariza , Zhu Dakai K. 
Published: April 2003

<i>Objectives:</i> Multivariate tests for linkage can provide improved power over univariate tests but the type I error rates and comparative power of commonly used methods have not previously been compared. Here we studied the behavior of bivariate formulations of the variance component (VC) and Haseman-Elston (H-E) approaches. <i>Methods:</i> We compared through simulation studies the bivariate H-E test with the unconstrained bivariate VC approach and with a VC approach in which the major-gene correlation is constrained to ±1. We also compared these methods to univariate methods. <i>Results:</i> Bivariate approaches are more powerful than univariate analyses unless the traits are very highly positively correlated. The power of the bivariate H-E test was less than the VC procedures. The constrained test was often less powerful than the unconstrained test. The empirical distributions of the bivariate H-E test and the unconstrained bivariate VC test conformed with asymptotic distributions for samples of 100 or more sibships of size 4. <i>Conclusions:</i> The unconstrained VC test is valuable for testing for preliminary linkages using multivariate phenotypes. The bivariate H-E test was less powerful than the bivariate VC tests.
Ecalectin as a T Cell-Derived Eosinophil Chemoattractant
Authors: Hirashima Mitsuomi 
Published: April 2003

In our previous papers, we have shown that human T cells produce a unique eosinophil chemotactic factor (ECF), termed Ecalectin, with a molecular weight of about 30–50 kD during interaction with BALL-1 (a B cell line) extracts, antigen or mitogen. A 1.6-kB cDNA was isolated from a human T cell-derived expression library that encodes Ecalectin. Ecalectin is a 36-kD protein consisting of 323 amino acids based on its deduced amino acid sequence. Ecalectin was found to be a variant of human galectin-9, a member of the growing family of animal lectins exhibiting affinity for β-galactosides. Recombinant Ecalectin, expressed in COS cells, exhibited potent ECF activity in vitro and in vivo in a dose-dependent manner but not chemotactic activity for neutrophils, lymphocytes, or monocytes. The finding that the Ecalectin transcript is present in various lymphatic tissues and that its expression increases substantially in antigen-activated peripheral blood mononuclear cells suggests that Ecalectin is an important T cell-derived regulator of eosinophil recruitment in allergic reaction sites.
Interferon-γ Increases CD62L Expression on Human Eosinophils
Authors: Momose T. , Okubo Y. , Horie S. , Takashi S. , Tsukadaira A. , Suzuki J. , Isobe M. , Sekiguchi M. 
Published: April 2003

<i>Background:</i> L-selectin (CD62L) and Mac-1 (CD11b/CD18) play a crucial role in the infiltration of eosinophils. However, changes in CD62L and CD11b expression on eosinophils after stimulation with cytokines were little studied. <i>Methods:</i> Eosinophils in peripheral blood of healthy volunteers were purified and cultured with interleukin-5 (IL-5), granulocyte/macrophage colony-stimulating factor (GM-CSF) or interferon-γ (IFN-γ). After up to 24 h incubation, CD62L and CD11b expression were examined using flow cytometry. The effects of dexamethasone (Dex), cycloheximide (CHX) or theophylline on CD62L expression on IFN-γ-stimulated eosinophils were also studied. <i>Results:</i> IL-5 or GM-CSF downregulated CD62L and upregulated CD11b expression on eosinophils after 30 min stimulation. Conversely, IFN-γ upregulated CD62L after 12 h stimulation in a time- and dose-dependent manner, and had no effect on CD11b expression. Dex, CHX or theophylline dose-dependently decreased CD62L expression on IFN-γ-stimulated eosinophils. <i>Conclusions: </i>IFN-γ is a particular cytokine which can increase CD62L expression on circulating or infiltrated human eosinophils. It is suggested that protein synthesis and intracellular cAMP participate in the increase in L-selectin expression on IFN-γ-stimulated eosinophils.
Adhesion to Fibronectin Regulates Expression of Intercellular Adhesion Molecule-1 on Eosinophilic Cells
Authors: Higashimoto Ikkou , Chihara Junichi , Kawabata Masaharu , Nakajima Shigenori , Osame Mitsuhiro 
Published: April 2003

<i>Background:</i> Adhesion molecules may play an important role in eosinophilic activation as well as adherence of extracellular matrix (ECM) including fibronectin (FN) in the inflamed focus. Tissue eosinophils expressed inter-cellular adhesion molecule-1 (ICAM-1, CD54), whereas peripheral blood eosinophils did not. The molecular mechanisms of ICAM-1 expression on eosinophils are not clear. We immunologically examined the effect of adherence to FN on the expression of adhesion molecules in an eosinophilic cell line (EoL-1). <i>Methods:</i> EoL-1 cells, suspended at a concentration of 2 × 10<sup>6</sup> cells/ml, were cultured at 37°C in BSA or FN-coated 24-well plates. After 4 h, the cells were removed by pipetting, and the expression of adhesion molecules was evaluated by immunofluorescence. <i>Results:</i> Adherence of EoL-1 to FN enhanced ICAM-1 (CD54) expression on EoL-1 (FN vs. BSA: 84.76 ± 17.90 vs. 43.73 ± 7.60, mean fluorescent intensity). Regarding other adhesion molecules on EoL-1 (CD11a, CD18, CD49d), the expression was not significantly augmented by adherence to FN. <i>Conclusions: </i>We concluded that the signal via adhesion of FN regulates the expression of ICAM-1 on eosinophilic cells. This study suggests that eosinophilic adhesion to ECM via adhesion molecules plays an important role in the pathogenesis of allergic inflammation through eosinophilic functional changes, including regulation of expressive adhesion molecules such as ICAM-1.
Mechanisms of Eosinophil Cationic Protein Release in the Serum: Role of Adhesion Molecules
Authors: Kato Yoshiko , Fujisawa Takao , Terada Akihiko , Iguchi Kosei , Kamiya Hitoshi 
Published: April 2003

<i>Background:</i> Measurement of eosinophil cationic protein (ECP) in serum has been utilized as a marker for allergic inflammation. The serum level of ECP represents the level found in vivo plus additional proteins released in vitro from peripheral blood eosinophils during the coagulation period. The mechanisms of release, however, are unclear. We investigated a possible involvement of adhesion molecules in the ECP release. <i>Materials and Methods:</i> Venous blood was drawn in the presence of EDTA from allergic donors. The blood was incubated with neutralizing monoclonal antibodies to CD18, CD11a, CD11b, CD29, CD49d, CD54, α<sub>4</sub>β<sub>7</sub>, or isotype-matched control antibodies, respectively, at 4°C for 30 min. Calcium gluconate (calcium) was then added to induce coagulation. The blood was further incubated for 90 min and centrifuged to obtain the serum. ECP in the serum was measured with RIA. In some experiments, purified eosinophils were incubated with plasma and calcium, then ECP in the supernatants was assayed. <i>Results:</i> ECP in the samples with calcium was significantly higher than in those without calcium. Purified eosinophils released ECP upon plasma coagulation. Anti-CD18, CD49d, and α<sub>4</sub>β<sub>7</sub> antibodies significantly suppressed ECP levels in the serum. <i>Conclusions:</i> These results suggest that ECP release in the serum is calcium and plasma coagulation-dependent and that cell adhesion through α<sub>L</sub>β<sub>2</sub>, α<sub>M</sub>β<sub>2</sub>, α<sub>4</sub>β<sub>1</sub> and α<sub>4</sub>β<sub>7</sub> integrins is at least in part responsible for ECP release.
Eosinophil-Adhesion-Inducing Activity Produced by Antigen-Stimulated Mononuclear Cells Involves GM-CSF
Authors: Nagata Makoto , Yamamoto Hideaki , Tabe Kazuaki , Sakamoto Yoshio , Matsuo Hiroshi 
Published: April 2003

<i>Background:</i> The initial step of eosinophil accumulation in allergic inflammation is adhesion of circulating eosinophils to vascular endothelial cells (EC). There is evidence that the adhesive property of circulating eosinophils is upregulated following antigen exposure. Although the exact mechanism remains to be established, cytokine(s) produced by antigen-stimulated mononuclear cells is (are) likely key factor(s). <i>Objective:</i> The objective of this study was to examine the factor(s) responsible for eosinophil adhesion and migration induced by the antigen-stimulated mononuclear cells obtained from atopic asthmatics. <i>Methods:</i> Peripheral blood mononuclear cells (PBMC) isolated from house-dust-mite-sensitive bronchial asthmatics were cultured for 96 h in the presence or absence of 1 µg/ml <i>Dermatophagoides farinae (Df)</i> antigen. Eosinophils were isolated from peripheral blood of healthy subjects. Eosinophil-adhesion-inducing activity in the culture supernatants of PBMC was examined by the ability to modify the adhesion of eosinophils to human pulmonary microvascular endothelial cells (HPMEC) in the presence or absence of anti-cytokine/chemokine antibodies. Eosinophil migration induced by the supernatants was also examined. <i>Results:</i> Eosinophil adhesion to HPMEC was significantly augmented by the supernatants of <i>Df</i>-stimulated PBMC, which was significantly inhibited by anti-GM-CSF, but not by anti-IL-5, anti-RANTES, or isotype-matched controls. On the other hand, eosinophil migration induced by the supernatants was inhibited by anti-GM-CSF and partly by anti-RANTES. <i>Conclusion:</i> Both eosinophil adhesion and migration induced by the antigen-stimulated PBMC involve GM-CSF. In contrast, RANTES is involved only in the eosinophil migration. These molecules may participate in the development of eosinophil accumulation at the allergic inflammation sites.
Increased Sensitivity to IL-4 in Patients with Allergic Bronchopulmonary Aspergillosis
Authors: Khan Seema , McClellan Jim S. , Knutsen Alan P. 
Published: April 2003

<i>Background:</i> Allergic bronchopulmonary aspergillosis (ABPA) is characterized by a heightened Th2 CD4+ T cell response to <i>Aspergillus fumigatus</i> allergens and a hyper-IgE state compared to atopic asthmatic and cystic fibrosis patients without ABPA. We hypothesized that one reason for this response is increased sensitivity to IL-4 in ABPA, resulting in increased expression of CD23 and CD86, leading to a positive amplification mechanism which increases Th2 CD4+ T cell responses. <i>Methods:</i> Peripheral blood mononuclear cells isolated from 10 ABPA, 9 atopic, and 8 nonatopic subjects and stimulated for 48 h with varying concentrations of rIL-4 ranging from 0.1 to 50 ng/ml. The percentages of CD23+ and CD86+ B cells and the number of CD23+ molecules on CD20+ and CD86+CD20+ B cells were quantified by flow cytometry. <i>Results:</i> Total serum IgE levels were elevated in ABPA patients compared to atopic and nonatopic controls. At day 0 prior to culture, CD23 molecules per CD20+ B cell were significantly elevated in ABPA patients compared to atopic and to nonatopic patients. CD23 molecules per CD20+ B cell in ABPA and atopic patients decreased after 48 h in culture without IL-4 added and were similar. With IL-4 stimulation, ABPA patients had significantly increased rates of CD23 expression per B cell compared to atopic and nonatopic subjects (p < 0.001). Furthermore, ABPA had significantly increased numbers of CD23+ molecules per B cell and CD86+ B cell following IL-4 stimulation compared to atopic and nonatopic patients. Both ABPA and atopic patients at day 0 prior to culture had increased expression of CD86+ and CD23+CD86+ B cells compared to nonatopic patients. After 48 h in culture without IL-4, the percentages of CD86+ and CD23+CD86+ B cells decreased in ABPA and atopic patients. After stimulation with IL-4, ABPA patients had significant upregulation of CD23+CD86+ B cells compared to atopic and nonatopic patients. Similarly, the number of CD23 molecules per CD86+CD20+ B cell was significantly upregulated following IL-4 stimulation in ABPA patients compared to atopic and to nonatopic subjects. <i>Conclusions:</i> This is the first study to demonstrate that ABPA patients have increased sensitivity to IL-4 stimulation compared to other atopic individuals, such that ABPA > atopic >> nonatopic patients. The B cells from ABPA patients were significantly more sensitive to IL-4 stimulation compared to atopic and nonatopic patients with upregulation of CD23 and CD86 expression. ABPA subjects had increased CD86+ and CD23+CD86+ B cell expression on day 0 prior to culture and with upregulation of CD23+ molecules on CD86+CD20+ B cells. IL-4 also stimulated upregulated CD86+ expression on B cells in atopic patients with little effect on nonatopic patients. This study supports the premise that IL-4, IL-4Rα and CD86 are central targets in the treatment of ABPA and atopic disease.
Roles of CD4+ and CD8+ T Cells in Adjuvant Activity of Diesel Exhaust Particles in Mice
Authors: Fujimaki Hidekazu , Ui Naoya , Ushio Hiroko , Nohara Keiko , Endo Tomohiko 
Published: April 2003

Through an imbalance in Th1 and Th2 cytokine profiles, diesel exhaust particles (DEP) are thought to induce Th2-dominated IgE and IgG1 production. However, the roles of CD4+ and CD8+ T-cell subtypes in the increased immune responses to antigen in mice exposed to DEP are unclear. In the present study, we investigated whether treatment with anti-CD4 or anti-CD8 mAb abrogated the adjuvant activity of DEP. On day –1 and day 1, each group of mice was injected intraperitoneally with anti-CD4, anti-CD8, or rat IgG (vehicle). On day 0, the mice were immunized with ovalbumin (OVA) or OVA plus DEP. After 3 weeks, each mouse was boosted with 10 µg of OVA alone. On day 7 after the first injection with OVA+DEP or OVA alone, the numbers of total, IA+, CD80+/IA+ and CD86+/IA+ cells in peritoneal exudate cells (PEC) were higher in OVA+DEP-immunized mice than in OVA-immunized mice. Depletion of CD8+ cells resulted in a modulation of the production of granulocyte-macrophage colony-stimulating factor, IL-12 and PGE<sub>2</sub> in peritoneal exudate fluid from OVA+DEP-immunized mice. On day 28, DEP injection markedly increased IL-4 production in the culture supernatants of spleen cells from CD4+ or CD8+-depleted mice. Depletion of CD8+ cells in OVA+DEP-immunized mice resulted in a decrease in IFN-γ production compared with that in OVA-immunized mice. Adjuvant activity of DEP was observed in anti-OVA IgE, anti-OVA IgG1, anti-OVA IgG3, and total IgE production. Depletion of CD4+ T cells abrogated the adjuvant effect of DEP on anti-OVA IgE, and anti-OVA IgG1 production in plasma. However, depletion of CD8+ T cell inhibited the upregulated anti-OVA IgG3 production. These findings suggest that DEP injection may affect not only the function of CD4+ cells but also that of CD8+ T-cell subsets to modulate the synthesis of proinflammatory cytokine in PEC and type-1 and type-2 cytokine production in spleens.
Identification of Allergen Fractions of Wheat Flour Responsible for Anaphylactic Reactions to Wheat Products in Infants and Young Children
Authors: Takizawa Takumi , Arakawa Hirokazu , Tokuyama Kenichi , Morikawa Akihiro 
Published: April 2003

Wheat is a food allergen which occasionally causes anaphylactic reactions exclusively in young children. There is very little knowledge of the clinical outcome in cases of food-related anaphylaxis and the differences in the allergenic protein components of food involved, comparing individuals who have suffered from an anaphylactic reaction with other individuals. The objectives of the present study were to examine the clinical features of 7 young children who had experienced anaphylactic reactions after ingesting wheat flour-containing products, and to analyze the allergens in wheat flour responsible for the anaphylactic symptoms. We measured the total IgE levels and the levels of IgE antibodies specific to wheat flour and performed IgE immunoblotting, comparing the sera from these children with sera from patients with atopic dermatitis. All sera from children who had experienced anaphylactic reactions were found to be positive for IgE specific to wheat. IgE immunoblotting revealed that 3 of these 7 children had sera showing reactivity to components of the salt-soluble protein fraction (16, 35–67 and 94 kD) and salt-insoluble protein-containing fraction (16, 38 and 70 kD) and 4 had no sera showing reactivity to components of the salt-soluble fraction. Patients with atopic dermatitis showed similar staining patterns. Various proteins in wheat flour could be allergens responsible for anaphylaxis and atopic dermatitis in infants or young children. Our findings suggest that these two clinically diverse allergic diseases do not necessarily represent responses to different allergenic proteins of wheat.
Maternal Serum CA125 and CA15-3 Antigen Levels in Normal and Pathological Pregnancy
Authors: Bon G.G. , Kenemans P. , Verstraeten A.A. , Go S. , Philipi P.A. , van Kamp G.J. , van Geijn H.P. , van Vugt J.M.G. 
Published: April 2003

<i>Objective:</i> To evaluate the value of maternal serum CA125 and CA15-3 concentrations for discriminating pathological from normal pregnancies. <i>Methods:</i> Serum samples from 120 women, in whom pregnancy outcome was pathological, i.e. spontaneous abortion, fetal death, intrauterine growth retardation, chromosomal and structural abnormalities, and (pre)eclampsia, were assessed for CA125 and CA15-3 and compared with levels found in 350 women with a normal pregnancy outcome matched for age and duration of pregnancy. <i>Results:</i> Maternal CA125 serum values were significantly higher in the first and the third trimester of pregnancy (median 23.0 and 21.0 U/ml; p < 0.00001 and p < 0.001, respectively), compared to those in the second trimester (median 14.0 U/ml), but not significantly different from those obtained in pathological pregnancies. Maternal serum CA15-3 values were significantly higher during the third trimester (median 26.0 U/ml) compared to the first and second trimester of pregnancy (median 14.0 and 15.0 U/ml; p < 0.0001); CA15-3 serum levels in normal and pathological pregnancies showed no significant difference. <i>Conclusion:</i> Maternal serum levels of CA125 are higher during the first and third trimester of pregnancy. CA15-3 maternal serum levels are higher during the third trimester compared to the first and second trimester. Maternal CA125 and CA15-3 serum levels showed no relation with a pathological outcome of pregnancy.
Persistent Detection of Alfa-Fetoprotein in the Vagina without Overt Preterm Premature Rupture of the Membranes
Authors: Kishida Tatsuro , Yamada Hideto , Furuta Itsuko , Kobayashi Noriko , Hirayama Emi K. , Ebina Yasuhiko , Cho Kazutoshi , Okuyama Kazuhiko , Fujimoto Seiichiro 
Published: April 2003

<i>Objectives:</i> The objectives of this study were to clinically characterize and seek risk factors for the patients with persistently detected alfa-fetoprotein (AFP) in the vagina without showing overt preterm premature rupture of the membranes (PROM). <i>Study Design:</i> The cohort study was undertaken using vaginal-cervical specimens collected consecutively between 21 and 40 weeks of gestation from 25 consenting women. Demographic, obstetric, neonatal, and laboratory data from patients with persistent positive results of AFP kit tests without showing overt leakage of amniotic fluid were compared to those from controls. Cervical levels of interleukin-1α, interleukin-1β, interleukin-6, interleukin-8, and granulocyte elastase were measured by immunoassay, and the uterine cervix was assessed by transvaginal ultrasonography. Statistical analysis involved Fisher’s exact test and Mann-Whitney U test. <i>Results:</i> The incidence of overt preterm PROM was significantly higher in the patients with persistently detected AFP in preterm (3 in 4 cases) than in controls (3 in 21 controls). At sampling before determination of persistently detected AFP, patients with persistently detected AFP had significantly higher cervical levels of interleukin-6 and significantly shorter cervical length than controls. <i>Conclusions:</i> Patients with persistently detected AFP in the vagina without showing overt preterm PROM have the risk of overt preterm PROM. Increased levels of interleukin-6 in cervical specimens and short cervical length may be risk factors for the occurrence of persistent detection of AFP in the preterm.
Morphological Differences between GPIb Antibody-Induced and Shear Stress-Induced Platelet Aggregates
Authors: Nomura Shosaku , Tandon Narendra N. , Nakamura Takashi , Kambayashi Junichi 
Published: April 2003

We determined the morphological differences between NNKY5-5-induced and shear stress-induced platelet aggregates using confocal laser scanning microscopy, and investigated the effects of cytokines on these aggregates. As shear stress was increased from low to high, many aggregates were generated. Some platelets and aggregates already exhibited PAC-1 binding at low shear stress. And finally, aggregates were clearly stained by PAC-1 at high shear stress. Low shear stress-induced aggregates included fibrinogen, but not von Willebrand factor (vWF). In contrast, high shear stress-induced aggregates included both fibrinogen and vWF. NNKY5-5-induced platelet aggregates exhibited both PAC-1 binding and fibrinogen, but vWF was not found in them. The aggregate formation in NNKY5-5-treated platelet-rich plasma (PRP) at low shear stress was clearly more pronounced than that in untreated PRP. In addition, aggregates in NNKY5-5-treated PRP formed after 6 min under low shear stress were clearly stained by PAC-1 and included fibrinogen, but vWF was not found in them. In order to investigate the effects of cytokines on platelet activation under NNKY5-5 stimulation, changes in light transmission and light scatter were assessed with an AG-10. G-CSF, IL-6 and thrombopoietin (TPO) each enhanced light scatter compared to control PRP, although IL-3, GM-CSF, erythropoietin and stem cell factor did not. In particular, TPO significantly enhanced the ‘%-T’ and ‘S-Max’ of NNKY5-5-treated PRP. TPO clearly enhanced the aggregate formation with high shear stress or NNKY5-5 stimulation. These results suggest that the glycoprotein Ib (GPIb)-unmediated aggregates can be formed with only fibrinogen, but the GPIb-mediated aggregates by vWF and fibrinogen synergistically. In particular, the latter is formed more firmly, and both aggregates are enhanced by TPO.
Antithrombotic Activity of Orally Administered Low Molecular Weight Heparin (Logiparin) in a Rat Model
Authors: Hiebert Linda M. , Ping Tilly , Wice Sandra M. 
Published: April 2003

Previous studies in rats demonstrated that orally administered, unfractionated bovine lung heparin is absorbed and has a dose-dependent antithrombotic effect. The objective of this study was to determine if an oral low molecular weight heparin had a similar antithrombotic effect in the same model. Thrombosis was induced in rats by application of 10% formalin in 65% methanol to the exposed jugular vein. Immediately following, saline, unfractionated heparin (3.3–60 mg/kg) or the low molecular weight heparin, Logiparin (0.025–15 mg/kg; 20–30 rats per group) was placed in the stomach and 4 h later the jugular vein was inspected for a thrombus. Compared to saline, oral Logiparin reduced the incidence of thrombosis at all doses with a dose-dependent effect suggested. A significant increase was observed in the activated partial thromboplastin time and in plasma heparin concentrations, determined by Accuclot<sup>TM</sup> Heptest<sup>®</sup> and anti-factor Xa chromogenic assay for rats given oral Logiparin versus saline. A dose-dependent increase in plasma heparin concentration was observed when estimated by the anti-Xa chromogenic assay. Heparin was recovered in 9% of aortic endothelial samples when ≧0.8 mg/kg Logiparin was administered. A 50% reduction in thrombosis was observed at 0.1 mg/kg for oral Logiparin versus 7.5 mg/kg for unfractionated bovine lung heparin indicating that oral Logiparin is an effective antithrombotic agent at doses lower than unfractionated heparin. Orally administered low molecular weight heparin may be useful for the prevention and treatment of thrombosis.