Immunoassay method for the determination of immunoglobulin G using bacterial magnetic particles.

We have developed a novel immunoassay method using bacterial magnetic particles for the determination of immunoglobulin G (IgG). Fluorescein isothiocyanate (FITC) conjugated anti IgG-bacterial magnetic particles were prepared. The fluorescence quenching caused by agglutination of FITC-anti IgG antibody-bacterial magnetic particle conjugates was measured by using a fluorescence spectrophotometer. The aggregates based on specific immunoreaction were separated by a gelatin solution. The aggregation of bacterial magnetic particle conjugates was enhanced by application of a magnetic field. The relative fluorescence intensity correlated linearly with a concentration of IgG in the range 0.5-100 ng/mL.     

The first specific antibody against cytotoxic polyacetylenic alcohol, panaxynol

Antitumor polyacetylenic alcohol, panaxynol, was isolated and purified from a powder of the root of Panax ginseng C.A. Meyer. Panaxynol inhibited the growth of various kinds of cultured tumor cell lines in a dose-dependent manner. In this paper we demonstrated the first specific antibody production against panaxynol. Anti-panaxynol antibody was elicited in rabbits by immunization with panaxynol hemisuccinate-bovine serum albumin conjugate (panaxynol hemisuccinate-BSA conjugate). An enzyme immunoassay (EIA) for the determination of panaxynol was established using a double-antibody technique. The EIA was highly specific against panaxynol although the antibody showed a minimal cross-reactivity with other types of polyacetylenic alcohol, i.e. panaxydol (12.0%) and panaxytriol (0.77%). Panaxynol at a concentration as low as 6.4 ng/ml can be detected. Using this assay we reconfirmed the rapid consumption of panaxynol by target tumor cells in an in vitro-culture system. The anti-panaxynol antibody may be a valuable tool for studies of the biological properties of polyacetylenic compounds.     

Antiproliferative constituents in umbelliferae plants. IV. Constituents in the fruits of Anthriscus sylvestris Hoffm

The constituents in the fruit of Anthriscus sylvestris Hoffm. were investigated, and four lignans [deoxypodophyllotoxin, morelensin, (-)-deoxypodorhizone, and (-)-hinokinin], one phenylpropanoid [1-(3',4'-dimethoxyphenyl)-1 xi-hydroxy-2-propene], two phenylpropanoid esters [3',4'-dimethoxycinnamyl (Z)-2-angeloyloxymethyl-2-butenoate and 3',4'-dimethoxycinnamyl (Z)-2-tigloyloxymethyl-2-butenoate], and one polyacetylenic compound (falcarindiol) were isolated. Their antiproliferative activity against MK-1, HeLa and B16F10 cell lines is reported.     

A 20 GHz 8 bit multiplexer IC implemented with 0.5 μm WNx/W-gate GaAs MESFET's

An ultrahigh-speed 8 bit multiplexer (MUX) has been developed for future-generation optical-fiber communication systems having a data rate of 20 Gb/s. This IC was fabricated using a 0.5 μm WN_x/W-gate GaAs MESFET process based on optical lithography, ion implantation, and furnace annealing for good reproducibility and high throughput. The WN_x/W bilayer gate has a low sheet resistance, improving the circuit high frequency performance. To attain 20 GHz operation, advanced circuit techniques for the source-coupled FET logic (SCFL) were introduced. A cross coupled source-follower (CCSF) was developed mainly for the highest speed buffers to enhance the bandwidth. The first-stage T-type flip-flop was designed with optimization techniques and operated up to 21.1 GHz     

Restriction fragment length polymorphism of genes of the alpha 2(XI) collagen, bone morphogenetic protein-2, alkaline phosphatase, and tumor necrosis factor-alpha among patients with ossification of posterior longitudinal ligament and controls from the Japanese population

STUDY DESIGN: The present study analyzed the restriction fragment length polymorphism patterns of alpha 2(XI) collagen, bone morphogenetic protein-2, alkaline phosphatase, and tumor necrosis factor-alpha genes in patients with ossification of the posterior longitudinal ligament. This study investigates the genetic polymorphism of bone-induced factors in patients with ossification of the posterior longitudinal ligament and compares it with healthy control subjects. OBJECTIVES: To clarify the genetic markers linked to ossification of the posterior longitudinal ligament. SUMMARY OF BACKGROUND DATA: Ossification of the posterior longitudinal ligament is a genetic disease associated with abnormal calcium metabolism involving the posterior longitudinal ligament. Previous genetic studies have not identified the pathologic mechanism of ossification of the posterior longitudinal ligament. Histopathologic studies of ossification of the posterior longitudinal ligament and the animal model, the spinal hyperostotic mouse, have revealed an increase in Type XI collagen and bone morphogenetic protein-2 expression. METHODS: Eighteen Japanese patients with ossification of the posterior longitudinal ligament and 51 healthy, unrelated control subjects were investigated for the restriction fragment length polymorphism patterns of COL11A2, bone morphogenetic protein-2, alkaline phosphatase, and tumor necrosis factor-alpha, genes with various restriction endonucleases. RESULTS: The gene frequencies of COL11A2 obtained with BamHl (10.0 kb fragment) and HindIII (19.0 kb fragment) observed in patients with ossification of the posterior longitudinal ligament were higher compared with control subjects (0.43 and 0.14, respectively). These differences were statistically significant (BamHl P = 0.018; Hindlll P = 0.046). Two new restriction fragment length polymorphism patterns were detected of the bone morphogenetic protein-2 gene with Mspl and Taql and one already known restriction fragment length polymorphism pattern of the tumor necrosis factor-alpha gene with Ncol. However, they were not significantly different from the control subjects. CONCLUSIONS: Seven restriction fragment length polymorphisms of COL11A2 gene were identified. Two of them (BamHl, 10.0/10.0 kb genotype; HindIII, 19.0/19.0 kb genotype) were significantly different in patients with ossification of the posterior longitudinal ligament.     

Negative regulation by interleukin-3 (IL-3) of mouse early B-cell progenitors and stem cells in culture: transduction of the negative signals by betac and betaIL-3 proteins of IL-3 receptor and absence of negative regulation by granulocyte-macrophage colony-stimulating factor

The receptors for interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-5 share a common signaling subunit betac. However, in the mouse, there is an additional IL-3 signaling protein, betaIL-3, which is specific for IL-3. We have previously reported that IL-3 abrogates the lymphoid potentials of murine lymphohematopoietic progenitors and the reconstituting ability of hematopoietic stem cells. We used bone marrow cells from betac- and betaIL-3-knock-out mice to examine the relative contributions of the receptor proteins to the negative regulation by IL-3. First, we tested the effects of IL-3 on lymphohematopoietic progenitors by using lineage-negative (Lin-) marrow cells of 5-fluorouracil (5-FU)-treated mice in the two-step methylcellulose culture we reported previously. Addition of IL-3 to the combination of steel factor (SF, c-kit ligand) and IL-11 abrogated the B-lymphoid potential of the marrow cells of both types of knock-out mice as well as wild-type mice. Next, we investigated the effects of IL-3 on in vitro expansion of the hematopoietic stem cells. We cultured Lin-Sca-1-positive, c-kit-positive marrow cells from 5-FU-treated mice in suspension in the presence of SF and IL-11 with or without IL-3 for 7 days and tested the reconstituting ability of the cultured cells by transplanting the cells into lethally irradiated Ly-5 congenic mice together with "compromised" marrow cells. Presence of IL-3 in culture abrogated the reconstituting ability of the cells from both types of knock-out mice and the wild-type mice. In contrast, addition of GM-CSF to the suspension culture abrogated neither B-cell potential nor reconstituting abilities of the cultured cells of wild-type mice. These observations may have implications in the choice of cytokines for use in in vitro expansion of human hematopoietic stem cells and progenitors.     

Thermal buckling of cross-ply laminated composite and sandwich plates according to a global higher-order deformation theory

A two-dimensional global higher-order deformation theory is presented for thermal buckling of cross-ply laminated composite and sandwich plates. By using the method of power series expansion of continuous displacement components, a set of fundamental governing equations which can take into account the effects of both transverse shear and normal stresses is derived through the principle of virtual work. Several sets of truncated Mth-order approximate theories are applied to solve the eigenvalue problems of a simply supported multilayered plate. Modal transverse shear and normal stresses can be calculated by integrating the three-dimensional equations of equilibrium in the thickness direction, and satisfying the continuity conditions at the interface between layers and stress boundary conditions at the external surfaces. Numerical results are compared with those of the published three-dimensional layerwise theory in which both in-plane and normal displacements are assumed to be C⁰ continuous in the continuity conditions at the interface between layers. Effects of the difference of displacement continuity conditions between the three-dimensional layerwise theory and the global higher-order theory are clarified in thermal buckling problems of multilayered composite plates.     

Strength of spline joints assembled by forming

In this paper, a new assembling method of spline joints that enables tight fitting in a simple manner by allowing slight plastic deformation at the spline teeth was introduced. Experiments were carried out for the spline joints of medium carbon steel varying the overlap zone between the male and the female spline teeth. Axial joining strength was increased with increase in the overlap length due to the residual compressive stress by forming. The joint by the proposed method also showed higher torsional strength than the conventional joint. Improvement in the torsional strength was explained based on the deformation and hardness distribution around the spline teeth. With respect to the shape of overlap zone, better results were obtained when using the specimen having a uniform overlap length along the axial direction.