Phylogeny of the Eurasian freshwater turtles of the genus Mauremys Gray 1869 (Testudines), with special reference to a close affinity of Mauremys japonica with Chinemys reevesii

Phylogenetic relationships of the freshwater turtles of the genus Mauremys and representatives of several other batagurid genera were inferred from variations in 863 base positions of mitochondrial 12S and 16S rRNA genes. Results strongly suggested the non-monophyly of Mauremys by indicating the closest affinity of Mauremys japonica with Chinemys reevesii , the type species of the genus Chinemys . Recent morphological analyses of the batagurid genera suggested that Mauremys is a basal stock of the subfamily Geoemydinae, whereas Chinemys is a member of the subfamily Batagurinae as supported by several putative synapomorphs with other batagurine genera. It is thus probable that the morphological character states used to define Mauremys actually represent symplesiomorphy, and that morphological character states shared between Chinemys and other batagurine genera have resulted from convergence. Also, our results did not support a sister-group relationship between Mauremys annamensis and Mauremys mutica , which has been implicitly or explicitly assumed by a number of previous authors on the basis of morphological data. Instead, M. annamensis was indicated to be closest to Mauremys i versoni , a species assumed to be most divergent among the East Asian Mauremys by previous authors.     

Seasonal changes in abundance and shifts in dominance of life history stages of the carrageenophyte Sarcothalia crispata (Rhodophyta, Gigartinales) in south central Chile

The population dynamics of the carrageenophyte Sarcothalia crispatais described from subtidal beds at two localities in south-central Chile. Seasonal fluctuations in total density and biomass were not evident. Frondswere identified to phase by the presence of reproductive structures and theresorcinol reaction. The monthly changes in abundance of each kind offrond were determined. Permanent gametophytic or sporophyticdominance was not evident: the more exposed site showed a seasonal shiftfrom sporophytic dominance in summer to gametophytic dominance inwinter, whereas the more protected site showed an interannual shift fromgametophytic to sporophytic dominance. The differences between localitiesand years suggest a very local population dynamics with large contributionof self-seeding to the maintenance of the S. crispata beds.     

Enzyme specificity under dynamic control II: Principal component analysis of α-lytic protease using global and local solvent boundary conditions

The contributions of conformational dynamics to substrate specificity have been examined by the application of principal component analysis to molecular dynamics trajectories of alpha-lytic protease. The wild-type alpha-lytic protease is highly specific for substrates with small hydrophobic side chains at the specificity pocket, while the Met190-->Ala binding pocket mutant has a much broader specificity, actively hydrolyzing substrates ranging from Ala to Phe. Based on a combination of multiconformation analysis of cryo-X-ray crystallographic data, solution nuclear magnetic resonance (NMR), and normal mode calculations, we had hypothesized that the large alteration in specificity of the mutant enzyme is mainly attributable to changes in the dynamic movement of the two walls of the specificity pocket. To test this hypothesis, we performed a principal component analysis using 1-nanosecond molecular dynamics simulations using either a global or local solvent boundary condition. The results of this analysis strongly support our hypothesis and verify the results previously obtained by in vacuo normal mode analysis. We found that the walls of the wild-type substrate binding pocket move in tandem with one another, causing the pocket size to remain fixed so that only small substrates are recognized. In contrast, the M190A mutant shows uncoupled movement of the binding pocket walls, allowing the pocket to sample both smaller and larger sizes, which appears to be the cause of the observed broad specificity. The results suggest that the protein dynamics of alpha-lytic protease may play a significant role in defining the patterns of substrate specificity. As shown here, concerted local movements within proteins can be efficiently analyzed through a combination of principal component analysis and molecular dynamics trajectories using a local solvent boundary condition to reduce computational time and matrix size.     

Insulin-like growth factor I receptors in neuronal and glial cells. Characterization and biological effects in primary culture

Primary cultures of neuronal and glial cells from 1-day-old neonatal rats contain high affinity receptors for insulin-like growth factor I (IGF-I). The IC50 for displacement of 125I-IGF-I binding by unlabeled IGF-I was 3 nM for neuronal cells and 4 nM for glial cells. Unlabeled insulin was 20-50 times less potent. Apparent molecular mass of the alpha subunits of the IGF-I receptor was 125 kDa in neuronal and 135 kDa in glial cells. IGF-I induced autophosphorylation of the IGF-I receptor beta subunit in lectin-purified membrane preparations in a dose-dependent manner. The major phosphoamino acid of the beta subunit in both cell types was tyrosine in the IGF-I-stimulated state and serine in the basal state. Apparent molecular mass of the beta subunits of the IGF-I receptors was 91 kDa for neuronal and 95 kDa for glial cells. Tyrosine kinase activity of the IGF-I receptors was demonstrated by IGF-I-induced phosphorylation of the exogenous substrate poly(Glu, Tyr) 4:1 in both cell types. IGF-I had no effect on 2-deoxyglucose uptake in neuronal cells. In contrast, in glial cells, IGF-I stimulated 2-deoxyglucose uptake at very high doses, presumably acting via the insulin receptor. The effect of IGF-I as a neurotrophic growth factor in both neuronal and glial cells was demonstrated by its stimulation of [3H]thymidine incorporation. These findings suggest the IGF-I is an important growth factor in nervous tissue-derived cells.     

Calcium concentration in brains from multiple sclerosis patients

Calcium (Ca) concentrations were studied in the central nervous system (CNS) of four patients with definite multiple sclerosis (MS, average age 49 yr) and five controls (average age 68 yr). The Ca concentration was determined by neutron activation analysis in autopsy samples taken from the 26 subanatomical regions of CNS tissues. Although the mean Ca concentration in the 26 CNS regions combined was higher in the four MS patients than in the controls, the content of white matter was lower. Whether or not this significantly lower Ca concentration found in the white matter of MS patients plays an important role in the demyelinating process remains unclear, although that lower concentration seems not be age dependent, but MS specific.