Transforming Growth Factor-α (TGFA): Genomic Structure, Boundary Sequences, and Mutation Analysis in Nonsyndromic Cleft Lip/Palate and Cleft Palate Only

Transforming growth factor-α (TGFA) has been proposed as a candidate gene in the etiology of nonsyndromic cleft lip with or without cleft palate (NS-CL/P) and of nonsyndromic cleft palate only (NS-CPO). Biologic support for a role of TGFA arises from its presence at high levels in the epithelial tissue of the medial edge of the palatal shelves at the time of shelf fusion in mice. Genetic support for the role of TGFA in clefting comes from the reported association of TGFA alleles with human NS-CPO and NS-CL/P. In this study we report the sequence and structure of human genomic TGFA and the search for causal TGFA mutations in 250 individuals with NS-CL/P or NS-CPO by conformational analysis of the coding sequence, splice junctions, and a portion of the 3′ untranslated region strongly homologous between human and mouse. We confirm that human TGFA is composed of six exons and here report several new sequence substitutions and their frequencies. Five variants in conserved segments may represent rare causes for clefting in humans and provide support for the role of TGFA in facial morphogenesis.     

Synthetic studies on 2'-substituted-4'-thiocytidine derivatives as antineoplastic agents.

As potential antineoplastic agents, we have synthesized 4'-thioFAC and 4'-thiocytarazid by developing an alternative synthetic method. 4'-ThioFAC showed potent antineoplastic activities in vivo as well as in vitro.     

Synthesis of 4'-substituted nucleosides and their biological evaluation.

4'-C-Ethynyl-beta-D-arabino- and 4'-C-ethynyl-beta-D-2'-deoxy-ribo-pentofuranosyl pyrimidines were synthesized. Most of these 4'-ethynyl nucleosides showed interesting biological activities.     

Morphologies and population genetic structures of the eight-barbel loach of the genus Lefua on southern Sakhalin

Ichthyological Research - Coldwater, primary freshwater, fish such as the eight-barbel loach, Lefua nikkonis, are thought to have colonised Hokkaido from the continental Far East via Sakhalin...     

Antitumor Activity of a Pterin Deaminase

A variety of bacteria and yeasts were examined for activities of biotin biosynthetic enzymes, including pimelyl-CoA synthetase, 7-keto-8-aminopelargonic acid (KAPA) synthetase, 7,8-diaminopelargonic acid (DAPA) aminotransferase and dethiobiotin (DTB) synthetase. Among the strains tested, only Bacillus sphaericus, a DTB producer, showed significant activities for all four enzymes. The bacterium also exhibited high activity of biotin synthesis from DTB in an intact cell system. Using cell-free extract and intact cells, some properties of DAPA aminotransferase, DTB synthetase and biotin synthesizing reaction were examined.

Based on these results of enzyme activities DTB productivity of B. sphaericus was discussed.     

Characterization of three electrophoretic variants of human erythrocyte triosephosphate isomerase found in Japanese

Three new electrophoretic variants of human erythrocyte triosephosphate isomerase (TPI) have been partially purified and compared with the normal isozyme with respect to stability, kinetics, and immunological properties. TPI 2HR1, an anodally migrating variant, was less stable than normal in guanidine denaturation and thermodenaturation tests, although it exhibited normal kinetic properties. The level of enzyme activity in erythrocytes from the proband with the phenotype TPI 1-2HR1 was about 60% of the normal mean. The variant allozyme TPI 2NG1, an anodally migrating allozyme associated with normal activity, was very thermolabile at 55 and 57°C. It was also much more labile than normal in stability tests in buffers at pH 5 and pH 10, although it exhibited normal kinetic and immunological properties. TPI 4NG1, a cathodally migrating variant associated with normal activity and normal kinetic as well as immunological properties, was more stable than normal in pH 5 buffer. Family studies demonstrated that the unique characteristics of these variants are genetically transmitted. In two-dimensional electrophoresis of purified isozymes the variant subunits were separated from the normal in the pI axis. However, there is no difference between the variants and the normal in the molecular weight axis, suggesting that the variants result from single amino acid substitutions.     

Intra- and extracellular localization of hyaluronic acid and proteoglycan constituents (chondroitin sulfate, keratan sulfate, and protein core) in articular cartilage of rabbit tibia.

To demonstrate the intra- and extracellular localization of hyaluronic acid (HA) in articular cartilage of the rabbit tibia, biotinylated HA binding region, which specifically binds to the HA molecule, was applied to the tissue. In comparison with the localization of HA, that of chondroitin sulfate (CS), keratan sulfate (KS), and the protein core (PC) of the proteoglycan was examined by immunohistochemistry. Strong positive staining for HA was detected in chondrocytes located in the transition between the superficial and middle zones of the tissue. Pre-treatment with chondroitinase ABC, keratanase II, or trypsin enhanced the stainability for HA in peri- and intercellular matrices. Immunohistochemistry with or without enzymatic pre-treatment demonstrated that immunoreactivity for CS, KS, and PC was distinctly discerned in chondrocytes and in the extracellular matrix located in the middle and deep zones. In particular, the immunoreactivity for KS and PC was augmented by pre-treatment with chondroitinase ABC not only in chondrocytes but in the extracellular matrix located in the middle and deep zones. Microbiochemical analysis corresponded well with histochemical and immunohistochemical results. These results suggest that HA is abundantly synthesized and secreted in chondrocytes located in the transition between the superficial and middle zones.