Neonatal administrations of a vasopressin analog (DDAVP) and hypertonic saline enhance learning behavior in rats

Groups of newborn Wistar rats received daily 1-desamino-8-D-arginine-vasopressin (DDAVP), oxytocin (OXT), hypertonic saline or normal saline for 14 days from day 1 to day 14 of life. One or three months later they were trained in a maze for brightness discrimination (BD). A group of untreated adult male rats received posttrial DDAVP or normal saline for brightness discrimination. Subsequently all the retentions of BD were tested after one month. We found that the neonatal treatments with both DDAVP and hypertonic saline facilitated acquisition and subsequent maintenance of brightness discrimination in immature and mature rats, and also that posttreatment with DDAVP enhanced retention of BD in adult rats. Oxytocin and normal saline had no effect on these parameters. The results are interpreted as showing that endogenous AVP and its synthetic analog enhance the development and adult function of central neural substrates involved in learning behaviors.     

Cholesterol myristate suppresses the apoptosis of mesenchymal stem cells via upregulation of inhibitor of differentiation

To identify small molecules that suppress the apoptosis of mesenchymal stem cells (MSCs) is promising for stem cell therapy. We recently showed that bone morphogenetic protein 4 (BMP4) signalling involves the effect of cholesterol myristate on the proliferation of MSCs. The present study evaluated the effects of cholesterol myristate on the apoptosis of MSCs and the inhibitor of differentiation (Id1), target gene of BMP4 signalling. MSCs transfected by the Id1 promoter reporter construct, cholesterol myristate increases the activity of Id1 promoter. However, structurally related steroids such as cholesterol, β-sitosterol and cholesten-3-one, lack of the myristate, did not affect the activity of Id1 promoter, suggesting that myristate is essential for this effect. This effect depends on BMP signalling. Apoptosis analysis indicated that cholesterol myristate inhibited the apoptosis of MSCs induced by serum-free. Cholesterol myristate increases the expression of Id1 and its target gene bcl-x/l in MSCs treated with serum-free. Moreover, noggin, a BMP antagonist, reduced the anti-apoptotic effects of cholesterol myristate. Thus, this study aims to provide evidence that cholesterol myristate suppresses the apoptosis of MSCs via up-regulation of Id1. These findings can be applied for improving MSCs survival in stem-cell transplantation, bone-marrow transplantation, treatment of bone diseases such as osteoporosis and chemotherapy.     

beta-sheet structure formation of proteins in solid state as revealed by circular dichroism spectroscopy

Cross beta-sheet structure formation and abnormal aggregation of proteins are thought to be pathological characteristics of some neurodegenerative disorders. To investigate the novel structural transformation and aggregation, the solid-state secondary structures of some proteins and peptides associated in thin films were determined by circular dichroism spectroscopy. Insulin, lysozyme, DsbA protein, luciferase, and ovalbumin peptide fall into one group; they show no or slight structural rearrangement from solution to the solid state. Another group, including bovine serum albumin, ovalbumin, alpha-synuclein, and plasminogen activator inhibitor-1 (PAIRC) peptide, undergo structural transformation with an increase of beta-sheet structure in the solid state. The beta-sheet formation of PAIRC peptide may reflect the structural transformation of the serpin reactive center that is relevant to the inhibitor activity. The beta-sheet structure of alpha-synuclein in the solid state may correspond to the amyloid-like aggregates, which are implicated in the pathogenesis of some neurodegenerative diseases.     

Structural evidence for a functional role of human tissue nonspecific alkaline phosphatase in bone mineralization

The human tissue nonspecific alkaline phosphatase (TNAP) is found in liver, kidney, and bone. Mutations in the TNAP gene can lead to Hypophosphatasia, a rare inborn disease that is characterized by defective bone mineralization. TNAP is 74% homologous to human placental alkaline phosphatase (PLAP) whose crystal structure has been recently determined at atomic resolution (Le Du, M. H., Stigbrand, T., Taussig, M. J., Ménez, A., and Stura, E. A. (2001) J. Biol. Chem, 276, 9158-9165). The degree of homology allowed us to build a reliable TNAP model to investigate the relationship between mutations associated with hypophosphatasia and their probable consequences on the activity or the structure of the enzyme. The mutations are clustered within five crucial regions, namely the active site and its vicinity, the active site valley, the homodimer interface, the crown domain, and the metal-binding site. The crown domain and the metal-binding domain are mammalian-specific and were observed for the first time in the PLAP structure. The crown domain contains a collagen binding loop. A synchrotron radiation x-ray fluorescence study confirms that the metal in the metal-binding site is a calcium ion. Several severe mutations in TNAP occur around this calcium site, suggesting that calcium may be of critical importance for the TNAP function. The presence of this extra metal-binding site gives new insights on the controversial role observed for calcium.     

Genes regulating touch cell development in Caenorhabditis elegans

To identify genes regulating the development of the six touch receptor neurons, we screened the F(2) progeny of mutated animals expressing an integrated mec-2::gfp transgene that is expressed mainly in these touch cells. From 2638 mutated haploid genomes, we obtained 11 mutations representing 11 genes that affected the production, migration, or outgrowth of the touch cells. Eight of these mutations were in known genes, and 2 defined new genes (mig-21 and vab-15). The mig-21 mutation is the first known to affect the asymmetry of the migrations of Q neuroblasts, the cells that give rise to two of the six touch cells. vab-15 is a msh-like homeobox gene that appears to be needed for the proper production of touch cell precursors, since vab-15 animals lacked the four more posterior touch cells. The remaining touch cells (the ALM cells) were present but mispositioned. A similar touch cell phenotype is produced by mutations in lin-32. A more severe phenotype; i.e., animals often lacked ALM cells, was seen in lin-32 vab-15 double mutants, suggesting that these genes acted redundantly in ALM differentiation. In addition to the touch cell abnormalities, vab-15 animals variably exhibit embryonic or larval lethality, cell degenerations, malformation of the posterior body, uncoordinated movement, and defective egg laying.     

Occurrence and transferability of β-lactam resistance inEnterobacteriaceae isolated inChildren's University Hospital in Bratislava

Occurrence and transferability of β-lactam resistance in 30 multi-resistantEscherichia coli, Klebsiella spp.,Enterobacter spp.,Pantoea agglomerans, Citrobacter freundii andSerratia marcescens strains isolated from children between 0 and 3 years of age is presented. The strains were resistant to ampicillin (30), cefoxitin (22), cefotaxime (30), ceftriaxone (30), ceftazidime (30) and aztreonam (28), but susceptible to cefepime (30) and imipenem (26). Twenty-eight of 30 isolates possessed a transferable resistance confirmed by conjugation and isolation of 79–89-kb plasmids. The β-lactam resistance was due to production of β-lactamases and ceftazidime proved to be stronger β-lactamase inductor than ceftriaxone. Twenty-five clinical isolates expressed transferable extended spectrum β-lactamases, and chromosomally encoded AmpC β-lactamase.