Human Fc epsilon RIalpha-specific human single-chain Fv (scFv) antibody with antagonistic activity toward IgE/Fc epsilon RIalpha-binding

The alpha-chain of Fc epsilon RI (Fc epsilon RIalpha) plays a critical role in the binding of IgE to Fc epsilon RI. A fully human antibody interfering with this interaction may be useful for the prevention of IgE-mediated allergic diseases. Here, we describe the successful isolation of a human single-chain Fv antibody specific to human Fc epsilon RIalpha using human antibody phage display libraries. Using the non-immune phage antibody libraries constructed from peripheral blood lymphocyte cDNA from 20 healthy subjects, we isolated three phage clones (designated as FcR epsilon 27, FcR epsilon 51, and FcR epsilon 70) through two rounds of biopanning selection. The purified soluble scFv, FcR epsilon 51, inhibited the binding of IgE to recombinant Fc epsilon RIalpha, although both FcR epsilon 27 and FcR epsilon 70 showed fine binding specificity to Fc epsilon RIalpha. Since FcR epsilon 51 was determined to be a monomer by HPLC, BIAcore analysis was performed. The dissociation constant of FcR epsilon 51 to Fc epsilon RIalpha was estimated to be 20 nM, i.e., fortyfold lower than that of IgE binding to Fc epsilon RIalpha (K(d) = 0.5 nM). With these characteristics, FcR epsilon 51 exhibited inhibitory activity on the release of histamine from passively sensitized human peripheral blood mononuclear cells.     

Synthesis and absolute configuration of MQ-A3 [1-(14'-methylhexadecanoyl) pyrrolidine], a novel aliphatic pyrrolidine amide from the tropical convolvulaceous species

A novel pyrrolidine amide (MQ-A3) isolated from the tropical convolvulaceous species was synthesized in 5 steps by starting from commercially available 12-bromododecanol and (S)-2-methylbutylbromide. The absolute configuration of the natural product was confirmed by a comparison of the specific rotation values.     

Targeted disruption of LIG-1 gene results in psoriasiform epidermal hyperplasia

We have designed a chimeric promoter that can be stimulated by various pro-inflammatory mediators and so drive the expression of therapeutic genes under inflammatory conditions. The promoter has two parts, the [-247/+20] fragment of the human type IIA secreted phospholipase A2 gene promoter, which is stimulated by the pro-inflammatory cytokine interleukin-1beta (IL-1beta), and a double peroxisome proliferator-activated receptor response element that is activated by some eicosanoids and by non-steroidal anti-inflammatory drugs (NSAIDs). Transfection experiments using rabbit articular chondrocytes in primary culture showed that this chimeric promoter produced a low basal activity and was induced by NSAIDs, WY-14643, IL-1beta, and 15-deoxy Delta12,14 prostaglandin J2. The latter two compounds stimulated the promoter synergistically.     

Deer browsing reduces leaf damage by herbivorous insects through an induced response of the host plant

We aimed to demonstrate an indirect relationship between a mammalian herbivore (sika deer) and herbivorous insects on the induced responses of a shared host plant, Viburnum dilatatum. Field studies were conducted at three sites (i.e. two islands and one mainland) and within a deer exclusion area. One island, Kinkazan (Kz) Island, harbored a high density of deer while the other sites (controls) had no deer or very low densities of deer. The deer exclusion area had been established approximately 10years earlier on Kz. We collected leaves above the browsing line of the deer and measured leaf hardness and tannin concentration. Leaf damage by insects was used as a measure of insect abundance. Leaves collected at Kz were harder than those from one of the control sites and from inside the deer exclusion area, while no difference was detected among the other controls and inside the exclusion area. In contrast, the tannin concentration of leaves from Kz was lower than in leaves from the control site. Leaf damage by herbivorous insects was lower in Kz than the other study sites. In addition, hole-type leaf damage tended to be higher inside, rather than outside, the exclusion area. These results suggest the possibility that deer browsing increased leaf hardness, which exerted an indirect negative effect on the herbivorous insects utilizing the common host plant. To our knowledge, this is the first study to provide evidence of indirect negative effects between mammalian herbivores and herbivorous insects sharing a host plant.     

The first synthesis and antifungal activities of 9-methoxystrobilurin-type beta-substituted beta-methoxyacrylate

The first synthesis of 9-methoxystrobilurin-type beta-substituted MOAs was successfully achieved. A chiral oudemansin-type beta-substituted MOA was also synthesized utilizing Mukaiyama's asymmetric aldol reaction. Antifungal activities of the synthesized compounds against several representative fungi were examined by disk-diffusion assay. As a result, unique and superior antifungal properties of 9-methoxystrobilurin-type beta-substituted MOAs compared with those of oudemansin-type analogue were clearly revealed.     

Physical mapping of plasmid and cosmid clones in filamentous fungi by fiber-FISH

Fluorescence in situ hybridization to extended DNA fibers (fiber-FISH) serves as a powerful tool for direct physical mapping in plants and animals. Here, we show that fiber-FISH is useful for contig mapping as well as for estimating the physical distance between genetic markers in fungi. A five-cosmid contig from a chromosome of Nectria haematococca and four cloned genetic markers from a linkage map of Cochliobolus heterostrophus were chosen as models for the application of this technology. In N. haematococca, overlapping and non-overlapping clones were visually mapped on individual DNA fibers, confirming the results from conventional physical mapping perfectly. Fiber-FISH concomitantly indicated the gap size or the extent of overlap between two clones. In C. heterostrophus, the physical distance between the two pairs of genetic markers could be estimated from the microscopic measurements of the intervals. Chromosomal DNA isolated from a pulsed field gel was suitable for preparing the DNA fibers.     

Single nucleotide polymorphisms of thrifty genes for energy metabolism: evolutionary origins and prospects for intervention to prevent obesity-related diseases

The "thrifty" genotype and phenotype that save energy are detrimental to the health of people living in affluent societies. Individual differences in energy metabolism are caused primarily by single nucleotide polymorphisms (SNPs), some of which promote the development of obesity/type 2 diabetes mellitus. In this review, four major questions are addressed: (1) Why did regional differences in energy metabolism develop during evolution? (2) How do genes respond to starvation and affluence? (3) Which SNPs correspond to the hypothetical "thrifty genes"? (4) How can we cope with disease susceptibility caused by the "thrifty" SNPs? We examined mtDNA and genes for energy metabolism in people who live in several parts of Asia and the Pacific islands. We included 14 genes, and the SNP frequencies of PPAR gamma 2, LEPR, and UCP3-p and some other genes differ significantly between Mongoloids and Caucasoids. These differences in SNPs may have been caused by natural selection depending on the types of agriculture practiced in different regions. Interventions to counteract the adverse effects of "thrifty" SNPs have been partially effective.