Substrate specificity of Geotrichum candidum lipase preparations

Summary Lipases with different fatty acid specificity were produced byGeotrichum candidum depending on growth condition. The hydrolysis of olive oil was inhibited by glycerol tributyrate and was dependent on Ca-ions for running at maximal rate.     

Expression of a cytomegalovirus IE-1-factor VIII cDNA hybrid gene in transgenic mice

A construct containing the 5′ end of the human cytomegalovirus major immediate early gene fused to the human coagulation factor VIII cDNA was used to produce transgenic mice. Two out of five transgenic lines transcribed the construct. The expression was consistently seen in a limited number, of tissues and was highest in muscle tissues. This is in contrast to the almost ubiquitous activity demonstrated in earlier studies with the IE-1 enhancer/promoter. Human factor VIII protein was detected immunochemically in muscle tissues at levels several times higher than in human plasma.     

Identification of determinants that confer ligand specificity on the insulin receptor.

We have previously shown, using truncated soluble recombinant receptors, that substituting the 62 N-terminal amino acids of the alpha subunit from the insulin-like growth factor I receptor (IGFIR) with the corresponding 68 amino acids from the insulin receptor (IR) results in a chimeric receptor with an approximately 200-fold increase in affinity for insulin and only a 5-fold decrease in insulin-like growth factor I (IGFI) affinity (Kjeldsen, T., Andersen, A. S., Wiberg, F. C., Rasmussen, J. S., Sch?ffer, L., Balschmidt, P., M?ller, K. B., and M?ller, N. P. H. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 4404-4408). We demonstrate that these 68 N-terminal amino acids of the IR also confer insulin affinity on the intact IGFI holoreceptor both in the membrane-bound state and when solubilized by Triton X-100. Furthermore, this domain can be subdivided into two regions (amino acids 1-27 and 28-68 of the IR alpha subunit) that, when replacing the corresponding IGFIR sequences, increases the insulin affinity of truncated soluble receptor chimeras 8- and 20-fold, respectively, with only minor effects on the IGFI affinity. Within the latter of these two regions, we found that amino acids 38-68 of the IR, representing 13 amino acid differences from IGFIR, confer the same 20-fold increase in insulin affinity on the IGFIR. Finally, the amino acids from position 42 to 50 are not responsible for this increase in insulin affinity. We thus propose that at least two determinants within the 68 N-terminal amino acids of the insulin receptor are involved in defining the ligand specificity of the insulin receptor, and that one or a combination of the remaining seven amino acid differences between position 38 and 68 are involved in conferring insulin affinity on the insulin receptor.     

Three-year study of benthic algal spring bloom development in a small, Danish lowland stream

Seasonal development of benthic algae was studied over a three-year period in a small, nutrient-rich lowland stream to investigate inter-annual variation in the algal spring bloom and differences in algal biomass regulation on two different substrata: fine-grained sediments and stones. The algal spring bloom was initiated when irradiance at the sediment surface exceeded 7 mol photons m^-2 d^-1 and mean water velocity was concomitantly below the threshold for bed load transport in the stream. Large inter-annual and substratum-dependent differences in peak algal biomass were observed, thus suggesting that different parameters regulate algal biomass development on the two substrata. On fine-grained sediments algal biomass development was predominantly coupled to light availability, while on stony substrata algal composition and peak biomass might be affected by invertebrate grazing.     

Fine Structural Observations on the Extracellular Matrix (ECM) of Xenoturbella bocki Westblad, 1949

Fine structural observations on the extracellular matrix (ECM) and connective tissue system of the enigmatic vermiform animal Xenoturbella bocki have demonstrated a complex and interesting organization of the ECM. Most conspicuous is the subepidermal membrane complex (SMC), and the major part of the ECM is present in this structure, which consists of a limiting basal lamina on each side of a central thick filamentous layer, probably of a collagenous nature. Distinct anchoring filaments are found as part of the SMC. A basal lamina is also observed in connection with the gastrodermal cells. The mesoderm is represented by a loose, ill-defined parenchyma, without filaments. The SMC is discussed in relation to subepidermal basal matrices occurring in turbellarians and enteropneusts, the two groups to which a relationship to Xenoturbella previously has been suggested. Comparisons between the ECM in Xenoturbella and in turbellarians do not support the notion of a relationship between these groups. Conversely, the present study strengthens previous indications of distinct similarities between certain characteristics of the epidermis and the SMC present in both enteropneusts and in Xenoturbella.     

Ribosomal proteins: XLIII. In vivo assembly of Escherichia coli ribosomal proteins

Isolation of ribosomal precursors from Escherichia coli K12 is described. The RNA and protein content of the precursor particles was determined.One physiologically stable precursor was found for the 30 S subunit. The assembly scheme is as follows: p16 S RNA + 9 proteins → p30 S (“21 S” precursor) p30 S + 12 proteins → 30 S subunit where p is precursor.Each of the two precursors for the 50 S subunit, P₁50 S and p₂50 S (“32 S” and “43 S” precursors, respectively), contains p5 S + p23 S RNA's in a 1:1 molar ratio. The assembly scheme is as follows: p23 S RNA + p5 S RNA + 16 or 17 proteins → p₁50 S

In contrast to the p₂50 S precursor the p₁50 S precursor is not similar to any core particles, which were obtained by treating 50 S subunits with different concentrations of LiCl or CsCl.The precursors p30 S and p₂50 S can be converted into active 30 S and 50 S sub-units, respectively, by incubation at 42 °C in the presence of ribosomal proteins and under RNA methylating conditions.     

Cytological studies on the planarian neoblast

Summary The paper is a study of the cytology of the regeneration cells (neoblasts) in Planaria vitta.The morphology of the living cells has first been examined to provide a reference for an investigation of the fixed neoblasts as studied by ordinary cytological, cytochemical and electron microscopical technics.A rather selective staining method has been devised based on the strong basophilic properties of the scanty cytoplasm. The morphology of the fixed neoblasts and their distribution in the intact animal have been described, using this method.The marked cytoplasmic basophilia was found to be exclusively due to ribonucleic acid, and not to desoxyribonucleic acid or acid mucopolysaccharides.The cytoplasm contains moderate to considerable amounts of basic proteins. Tyrosine, cysteine/cystin, arginine, lysine and perhaps histidine were present, while tryptophan could not be demonstrated.No enzymes could be demonstrated apart perhaps from cytochrome oxidase.The mitochondria are small and inconspicuous and more or less evenly distributed throughout the cytoplasm. A Golgi apparatus could not be demonstrated.The electron microscopic picture is very characteristic, because of the high electron density of the cytoplasm. This density is the result of the presence of a great number of ribonucleoprotein granules. Most of the granules are free and only a minor part bound to the membranes of the endoplasmatic reticulum. The interesting features of the cell membrane are discussed in relation to the structure of the parenchyma.The cytochemical properties of the neoblast (RNA and sulfhydryl-groupcontaining protein) and the fine structure as revealed in the electron microscope characterize the neoblast as a morphogenetically active cell.