Iridoid glucosides in fouquieriaceae

From three Fouquieria sp. 12 iridoid glucosides were isolated and identified. Eight of these were structurally related to galioside (monotropein methylester), while four were hydroxy substitution products of deoxyloganin. In three cases the glucoside occurred together with the corresponding 10-O-acetate.     

Single-file diffusion through the Ca2+-activated K+ channel of human red cells

Summary We have examined the effect of internal and external pH on Na⁺ transport across toad bladder membrane vesicles. Vesicles prepared and assayed with a recently modified procedure (Garty & Asher, 1985) exhibit large, rheogenic, amiloridesensitive fluxes. Of the total²²Na uptake measured 0.5–2.0 min after introducing tracer, 80±4% (mean±se,n=9) is blocked by the diuretic with aK ₁ of 2×10^–8 m. Thus, this amiloridesensitive flux is mediated by the apical sodium-selective channels. Varying the internal (cytosolic) pH over the physiologic range 7.0–8.0 had no effect on sodium transport; this result suggests that variation of intracellular pHin vivo has no direct apical effect on modulating sodium uptake. On the other hand,²²Na was directly and monotonically dependent on external pH. External acidification also reduced the amiloride-sensitive efflux across the walls of the vesicles. This inhibition of²²Na efflux was noted at external Na⁺ concentrations of both 0.2 m and 53mm.These results are different from those reported with whole toad bladder. A number of possible bases for these differences are considered and discussed. We suggest that the natriferic response induced by mucosal acidification of whole toad urinary bladder appears to operate indirectly through one or more factors, presumably cytosolic, present in whole cells and absent from the vesicles.     

Asexual propagation and reproductive strategies in aquatic Oligochaeta

A great variety of asexual reproductive modes are known among aquatic oligochaetes. The main types of these modes are shortly described. Based upon observations on natural populations, the possible genetical and ecological implications of asexual reproduction are discussed. The following points are emphasized: (1) The often expressed expectations of a strong predominance of one particularly adaptive genotype is not born out. (2) In most cases, a number of genetically distinct clones are present in each population, and they show a strong differential distribution in heterogeneous environments, indicating an effective exploitation of the available resources. (3) Most cases of asexual propagation are reproductive strategies of their own and not escape mechanisms. (4) The mechanisms underlying asexual propagation are complex and involve many aspects of the life history. The great variety of types among aquatic oligochaetes offer particularly useful models for the study of these problems.     

Effects of T3 and rearing temperature on growth and skeletal myosin heavy chain isoform transition during early development in the salmonid Salvelinus alpinus (L.)

Rearing of 1-year-old Arctic charr (Salvelinus alpinus) at 12°C, as well as the administration of 50 or 75 mgT₃/kg feed, accelerated the neonatal to adult fast myosin heavy chain transition, but the effect of temperature was more dramatic than the effect of T₃ administration. The endogenous plasma levels of T₃ in charrs reared at 12°C were higher than those of analogous groups reared at natural temperature, which in the period under study was between 0.5 and 12°C. As in other species, T₃ seemed to play a role in the regulation of the neonatal to adult fast myosin isoform transition by down-regulating the levels of the neonatal and increasing the levels of an adult fast myosin heavy chain. Temperature seemed to accelerate this transition at least, but not only, by inducing an increase in the endogenous levels of T₃ in the Arctic charr.     

Recent palaeolimnology of three shallow Danish lakes

Recent eutrophication histories of three shallow lakes (mean depths <3 m) were studied using palaeolimnological methods. Freeze-cores were dated using ²¹⁰Pb, ¹³⁷Cs, ¹³⁴Cs and ²⁴¹Am. Resultant chronologies were problematical at two sites (Væng Sø and Vesterborg Sø) due to sediment-water interface mixing, indicated by uniform ¹³⁷Cs profiles over the surface 20–30 cm. Sediments at Langesø and Vesterborg Sø have a high carbonate content, which together with the high mixing rate have resulted in diatom dissolution below 30 cm at Vesterborg Sø. Diatom stratigraphy indicates relatively small biological changes at both Væng Sø and Langesø both lakes have been eutrophic for at least the last 150–200 years. Væng Sø is dominated by planktonic diatoms together with high percentages of benthic Fragilaria spp., and Langesø by planktonic diatoms, especially Cyclostephanos dubius and Stephanodiscus parvus.Epilimnetic phosphorus concentrations were inferred using weighted averaging; at Væng Sø the diatom-inferred TP (DI-TP) concentrations were close to observed values in the early 1980s, but failed to record the post-biomanipulation decrease to 55 g l^-1 after 1988, presumably due to the smoothing of the sediment record by resuspension and mixing. At Langesø two increases in DI-TP suggest a two-phase enrichment of the lake, initial eutrophication 1880, and hypertrophy after 1950. The recent DI-TP values are lower than the annual observed values, and reasons for this are discussed. Despite problems associated with sediment mixing, the sediment records of these shallow systems can still be used for monitoring and for environmental reconstructions.     

Glycan modification of a thermostable recombinant (1-3,1-4)-β-glucanase secreted fromSaccharomyces cerevisiae is determined by strain and culture conditions

High level biosynthesis and secretion of the thermostable hybrid (1-3,1-4)--glucanase H(A16-M) has been achieved inSaccharomyces cerevisiae by means of the yeast vacuolar endoprotease B promoter (PRB1_p) and theBacillus macerans (1-3,1-4)--glucanase signal peptide. The N-glycans present on the yeast-secreted H(A16-M), denoted H(A16-M)-Y, were released by endoglycosidase H, and identified by proton NMR spectroscopy to be a homologous series of Man_8-13GlcNAc₂, although only traces of Man₉GlcNAc₂ were found. Therefore, processing of N-glycans on H(A16-M)-Y is similar to that on homologous proteins. Most of the N-glycans (88%) were neutral while the remainder were charged due to phosphorylation. Site-directed mutagenesis of Asn to Gln in two of the N-glycosylation sequons, and subsequent analysis of the N-glycans on the yeast-secreted proteins together with analysis of the N-glycans from the individual sites of H(A16-M)-Y suggest the presence of steric hindrance to glycan modification by the glycans themselves. H(A16-M)-Y produced under control of either the yeast protease B or the yeast 3-phosphoglycerate kinase promoter, each in two differentSaccharomyces strains revealed a dependence of N-glycan profile on both strain and culture conditions. The extent of O-glycosylation was found to be nine mannose units per H(A16-M)-Y molecule. An attempt to identify the linkage-sites for the O-glycans by amino acid sequencing failed, suggesting non-stoichiometric or heterogeneous O-glycosylation. The possible modes in which N-glycans might contribute to resistance of H(A16-M)-Y to irreversible thermal denaturation are discussed with respect to structural information available for H(A16-M)-Y. Abbreviations: AMY,B. amyloliquefaciens (1-3,1-4)--glucanse; MAC,B. macerans (1-3,1-4)--glucanase H(A16-M), H(A36-M), H(A78-M),H(A107-M) and H(A152-M), hybrid (1-3,1-4)--glucanases containing 16, 36, 78, 107 and 152 N-terminal amino acids, respectively, derived from AMY with the remaining amino acids derived from MAC; similar enzyme abbreviations followed by Y, e.g. H(A16-M)-Y, denote the enzymes secreted from yeast cells; PCR, polymerase chain reaction; PGK_p, yeast 3-phosphoglycerate kinase promoter; PRB1_p, yeast protease B promoter; LB, Luria-Bertani medium; SC, minimal medium; CNBr, cyanogen bromide; Endo H_f, endoglycosidase H fusion protein; PNGase F, peptide:N-glycosidase F; HPAEC; high pH anion exchange chromatography; HVE, high voltage paper electrophoresis; CPY, yeast carboxypeptidase Y.     

High-resolution mapping by YAC fragmentation of a 2.5-Mb Xp22 region containing the human RS, KFSD and CLS disease genes

The disease loci for X-linked Retinoschisis (RS), Keratosis follicularis spinulosa decalvans (KFSD), and Coffin-Lowry syndrome (CLS) have been localized to the same, small region in Xp22 on the human X Chromosome (Chr). To generate a high-resolution map of the available contig in this area, we have used the YAC fragmentation vectors pBP108/ADE2 and pBP109/ADE2 and generated fragmented YACs from a 2.5-Mb YAC (y939H7) spanning the mentioned disease gene candidate regions. Forty-seven fragmented YACs were generated and analyzed, ranging in size from 170 kb to over 2400 kb. The resulting YAC fragmentation panel was used to construct a detailed restriction map of the region and has been used to bin clones and markers. As a deletion panel, it will present a valuable resource for further mapping. Received: 31 December 1996 / Accepted: 22 February 1997     

Mechanical properties of trabecular bone. Dependency on strain rate.

The effect of strain rate (epsilon) and apparent density (rho) on stiffness (E), strength (sigma u), and ultimate strain (epsilon u) was studied in 60 human trabecular bone specimens from the proximal tibia. Testing was performed by uniaxial compression to 5% specimen strain. Six different strain rates were used: 0.0001, 0.001, 0.01, 0.1, 1, and 10 s-1. Apparent density ranged between 0.23 and 0.59 g cm-3. Linear and non-linear regression analyses using strength, stiffness and ultimate strain as dependent variables (Y) and strain rate and apparent density as independent variables were performed using the following models: Y = a rho b epsilon c, Y = rho b(a + c epsilon; Y = (a + b rho)epsilon c, Y = a rho 2 epsilon c, E = a rho 3 epsilon c. The variations of strength and stiffness were explained equally well by the linear and the power function relationship to strain rate. The exponent was 0.07 in the power function relationship between strength and strain rate and 0.05 between stiffness and strain rate. The variation of ultimate strain was explained best using a power function relationship to strain rate (exponent = 0.03). The variation of strength and stiffness was explained equally well by the linear, power function and quadratic relationship to apparent density. The cubic relationship between stiffness and apparent density showed a less good fit. Ultimate strain varied independently of apparent density.