Rapid Screening of Antigenically Reactive Fragments of asi-Casein Using HPLC and ELISA

Screening of antigenically reactive fragments of aSi-casein (asx-CN), the major casein in bovine milk, was done by using HPLC and enzyme-linked immunosorbent assay (ELISA). BALB/c mice (6-week-old) were injected intraperitoneally with aSi-CN and complete Freund's adjuvant, and 14 days later, all the mice were boosted with as^CN and incomplete Freund's adjuvant. Twenty-one days after the 1st immunization, the mice were bled and antiserum was separated. Anti as^CN antibody fraction was obtained by precipitation from the antiserum with 50% saturated ammonium sulfate. aSi-CN was digested with trypsin and chymotrypsin, and 35 peptides were purified from the digests by reversed-phase HPLC with ODS (octadecylsilica) columns. Reactivity of peptides with the antibody were examined by ELISA. The solid phase in the wells of the polystyrene microtiter plate was coated with peptides, and the plate was successively incubated with anti aSj-CN antibody, conjugate of anti mouse immunoglobulin with alkaline phosphatase (ALP) and substrate of ALP. Two tryptic fragments (the residues 104-119 and 133-151) and three chymotryptic fragments (33-54, 105-121, and 174-199) were positive in an ELISA test. These five fragments would correspond to four antigenic sites. We could thus find antigenically reactive fragments of aSj-CN by the direct and simple detection of specific antigen-antibody interaction

Effects of phosphodiesterase inhibitors on secretions of human monokines

ABSTRACTThe purpose of this study was to evaluate the effect of newly developed selective phosphodiesterase (PDE) inhibitors, KF19514 (type l/IV) and cilostazol (type III), and theophylline on the secretions of tumor necrosis factor a (TNFα) and interleukin-1β (IL-1 β) from human peripheral monocytes stimulated by lipopolysaccha- ride (LPS). Human blood monocytes were incubated with LPS in the absence or presence of KF19514, cilostazol or theophylline. TNFα and IL-1in the cell- free supernatants were measured with enzyme-linked immunosorbent assay. KF19514 showed significant inhibition on the release of TNFα (% inhibition ± SEM was 82.8 ± 7.4% at 1 nmol/L) and IL-1 β (34.4 ± 7.5% at 10 (μmol/L). In addition, KF19514 inhibited the expression of TNFa mRNA. Cilostazol inhibited the release of TNFa significantly (60.2 ± 8.9% at 30 μmol/L) but not IL-1 β. Theophylline inhibited slightly but significantly the release of TNFa at a therapeutic concentration (1 7.4 ± 5.1% at 100 μmol/L). These results suggest that theophylline may not only have a bronchodilating action but also an anti-inflammatory property in the treatment of bronchial asthma, and that KF19514 may have an anti-inflammatory action on at least the transcriptional level

Development of a broadband superluminescent diode based on self-assembled InAs quantum dots and demonstration of high-axial-resolution optical coherence tomography imaging

We developed a near-infrared (NIR) superluminescent diode (SLD) based on self-assembled InAs quantum dots (QDs) and demonstrated high-axial-resolution optical coherence tomography (OCT) imaging using this QD-based SLD (QD-SLD). The QD-SLD utilized InAs QDs with controlled emission wavelengths as a NIR broadband light emitter, and a tilted waveguide with segmented electrodes was prepared for edge-emitting broadband electroluminescence (EL) spanning approximately 1–1.3 μm. The bandwidth of the EL spectrum was increased up to 144 nm at a temperature of 25 °C controlled using a thermoelectric cooler. The inverse Fourier transform of the EL spectrum predicted a minimum resolution of 3.6 μm in air. The QD-SLD was subsequently introduced into a spectral-domain (SD)-OCT setup, and SD-OCT imaging was performed for industrial and biological test samples. The OCT images obtained using the QD-SLD showed an axial resolution of ~4 μm, which was almost the same as that predicted from the spectrum. This axial resolution is less than the typical size of a single biological cell (~5 μm), and the practical demonstration of high-axial-resolution OCT imaging shows the application of QD-SLDs as a compact OCT light source, which enables the development of a portable OCT system

An Optically Dark GRB Observed by HETE-2: GRB 051022

GRB 051022 was detected at 13:07:58 on 22 October 2005 by HETE-2. The location of GRB 051022 was determined immediately by the flight localization system. This burst contains multiple pulses and has a rather long duration of about 190 seconds. The detections of candidate X-ray and radio afterglows were reported, whereas no optical afterglow was found. The optical spectroscopic observations of the host galaxy revealed the redshift z = 0.8. Using the data derived by HETE-2 observation of the prompt emission, we found the absorption N_H = 8.8 -2.9/+3.1 x 10^22 cm^-2 and the visual extinction A_V = 49 -16/+17 mag in the host galaxy. If this is the case, no detection of any optical transient would be quite reasonable. The absorption derived by the Swift XRT observations of the afterglow is fully consistent with those obtained from the early HETE-2 observation of the prompt emission. Our analysis implies an interpretation that the absorbing medium could be outside external shock at R ~ 10^16 cm, which may be a dusty molecular cloud.Comment: 6 pages, 2 figures, accepted for publication in PASJ lette

T-2 Toxin-induced Toxicity in Pregnant Mice and Rats

T-2 toxin is a cytotoxic secondary fungal metabolite that belongs to the trichothecene mycotoxin family. This mycotoxin is a well known inhibitor of protein synthesis through its high binding affinity to peptidyl transferase, which is an integral part of the ribosomal 60s subunit, and it also inhibits the synthesis of DNA and RNA, probably secondary to the inhibition of protein synthesis. In addition, T-2 toxin is said to induce apoptosis in many types of cells bearing high proliferating activity. T-2 toxin readily passes the placenta and is distributed to embryo/fetal tissues, which include many component cells bearing high proliferating activity. This paper reviews the reported data related to T-2 toxin-induced maternal and fetal toxicities in pregnant mice and rats. The mechanisms of T-2 toxin-induced apoptosis in maternal and fetal tissues are also discussed in this paper

HETE-2 Observations of the X-Ray Flash XRF 040916

A long X-ray flash was detected and localized by the instruments aboard the High Energy Transient Explorer II (HETE-2) at 00:03:30 UT on 2004 September 16. The position was reported to the GRB Coordinates Network (GCN) approximately 2 hours after the burst. This burst consists of two peaks separated by 200 s, with durations of 110 s and 60 s. We have analyzed the energy spectra of the 1st and 2nd peaks observed with the Wide Field X-Ray Monitor (WXM) and the French Gamma Telescope (FREGATE). We discuss the origin of the 2nd peak in terms of flux variabilities and timescales. We find that it is most likely part of the prompt emission, and is explained by the long-acting engine model. This feature is similar to some bright X-ray flares detected in the early afterglow phase of bursts observed by the Swift satellite.Comment: 9 pages, 4 figures, Accepted for publication in PAS

CD44 on blood eosinophils as a novel marker of bronchial asthma management

Bronchial asthma (BA) is characterized by infiltration of the respiratory tracts by eosinophils. A wide variety of adhesion molecules is involved in the binding of eosinophils to the vascular endothelium and their subsequent transmigration from the circulation to the airways, while little is known about CD44 expression on eosinophils. In the present study we introduce a novel staining combination with which surface markers on eosinophils can be analyzed without purification prior to staining and examined whether the expression of CD44 on peripheral blood eosinophils could be monitored as a marker of the control of BA. Staining of eosinophils with anti-CD16 and anti-VLA-4 monoclonal antibodies enabled us to delineate eosinophils as VLA-4highCDl6- cells from other leukocyte populations in the whole blood. CD44 was constitutively expressed on resting eosinophils and expression increased following cytokine-mediated activation. In all BA patients examined, CD44high eosinophils were enriched in sputum relative to peripheral blood, indicating that eosinophils in sputum were more activated than those in the blood. By comparing the extent of CD44 expression on blood eosinophils from poorly controlled and well-controlled asthma patients, we unexpectedly found that the density of CD44 expression is lower on blood eosinophils from the poorly controlled group. Thus, the extent of CD44 expression on blood eosinophils defined as VLA- 4highCD16- cells is a novel marker indicative of the management of BA