Tuning glycosidase inhibition through aglycone interactions: pharmacological chaperones for Fabry disease and GM1 gangliosidosis.

Competitive inhibitors of either α-galactosidase (α-Gal) or ß-galactosidase (ß-Gal) with high affinity and selectivity have been accessed by exploiting aglycone interactions with conformationally locked sp(2)-iminosugars. Selected compounds were profiled as potent pharmacological chaperones for mutant lysosomal α- and ß-Gal associated with Fabry disease and GM(1) gangliosidosis.

Serum thrombopoietin level and thrombocytopenia during the neonatal period in infants with Down's syndrome.

BACKGROUND: The pathogenesis of thrombocytopenia during the neonatal period in Down's syndrome (DS) infants remains unclear. OBJECTIVE: To elucidate kinetic changes of serum thrombopoietin (TPO) level and platelet count, and their correlation in DS neonates. STUDY DESIGN: Twelve DS infants (male/female: 7/5, term/late preterm: 10/2) born between 1997 and 2007 were included. Blood samples were serially collected during the neonatal period and serum TPO levels were determined in 44 sera using an enzyme-linked immunosorbent assay. RESULTS: Thrombocytopenia <150 x 10(9) per liter was observed in seven (58%) patients. In 12 DS patients, the median TPO value showed 2.86 fmol ml(-1) on day 0, rose to 4.64 fmol ml(-1) on day 2, and thereafter decreased to 4.30 fmol ml(-1) on day 5, 2.40 fmol ml(-1) on days 11-15, and 1.75 fmol ml(-1) on days 28-30. This kinetics parallels that in historical non-DS controls. In 35 pair sample analysis from 11 patients without transient myeloproliferative disease, TPO level inversely correlated with platelet count (r=-0.38, P=0.023). However, there was no significant difference in TPO concentrations between thrombocytopenic and non-thrombocytopenic DS individuals. CONCLUSIONS: This is the first study to describe the relationship between TPO level and platelet count in neonates with DS. Median TPO levels and their kinetic changes in DS neonates are comparable to those in non-DS controls. In contrast to earlier findings in several studies showing higher TPO concentrations in thrombocytopenic non-DS newborns than those in non-thrombocytopenic counterparts, the response of the TPO system to thrombocytopenia in DS during the neonatal period seems suboptimal.

Module-intron correlation and intron sliding in family F/10 xylanase genes.

Xylanases are classified into two families, numbered F/10 and G/11 according to the similarity of amino acid sequences of their catalytic domain (Henrissat, B., Bairoch, A., 1993. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 293, 781-788). Three-dimensional structure of the catalytic domain of the family F/10 xylanase was reported (White, A., Withers, S.G., Gilkes, N.R., Rose, D.R., 1994. Crystal structure of the catalytic domain of the beta-1,4-glycanase Cex from Cellulomonas fimi. Biochemistry 33, 12546-12552). The domain was decomposed into 22 modules by centripetal profiles (Go, M., Nosaka, M., 1987. Protein architecture and the origin of introns. Cold Spring Harbor Symp. Quant. Biol. 52, 915-924; Noguti, T., Sakakibara, H., Go, M., 1993. Localization of hydrogen-bonds within modules in barnase. Proteins 16, 357-363). A module is a contiguous polypeptide segment of amino acid residues having a compact conformation within a globular domain. Collected 31 intron sites of the family F/10 xylanase genes from fungus were found to be correlated to module boundaries with considerable statistical force (p values <0.001). The relationship between the intron locations and protein structures provides supporting evidence for the ancient origin of introns, because such a relationship cannot be expected by random insertion of introns into eukaryotic genes, but it rather suggests pre-existence of introns in the ancestral genes of prokaryotes and eukaryotes. A phylogenetic tree of the fungal and bacterial xylanase sequences made two clusters; one includes both the bacterial and fungal genes, but the other consists of only fungal genes. The mixed cluster of bacterial genes without introns and the fungal genes with introns further supports the ancient origin of introns. Comparison of the conserved base sequences of introns indicates that sliding of a splice site occurred in Aspergillus kawachii gene by one base from the ancestral position. Substrate-binding sites of xylanase are localized on eight modules, and introns are found at both termini of six out of these functional modules. This result suggests that introns might play a functional role in shuffling the exons encoding the substrate-binding modules.

The difference in endocochlear and endolymphatic sac d.c. potentials in response to furosemide and canrenoate as diuretics.

We investigated the effects of furosemide, a loop diuretic, and canrenoate, an aldosterone antagonist, on the endocochlear potential (EP) and the endolymphatic sac potential (ESP) in the guinea pig. Furosemide produced no significant change in the ESP at a dose of 100 mg/kg after an intravenous infusion for 20 min. However, this dose decreased the EP to a negative level. Canrenoate produced no significant change in the EP at an intravenous dose of 300 mg/kg for 20 min, but it did decrease the ESP. The differences in the EP and ESP in the response to the diuretics indicate a dissimilarity of the origin of both d.c. potentials in the endolymphatic space.