Post translational modifications of recombinant human Papillomavirus type 6b major capsid protein

We have determined the post-translational modifications of the major capsid protein, L1 of human papillomavirus (HPV) type 6b. Since this virus cannot be cultured in the laboratory to obtain sufficient material for a study, a recombinant L1 protein produced in a vaccinia virus expression system was used in this investigation. Our results show that this protein is phosphorylated at serine residues and is also glycosylated. No myristoylation or palmitoylation was detected. The fraction of L1 protein incorporated into virus-like particles was not glycosylated. Since recombinant L1 protein is a potential human vaccine candidate, knowledge of the post-translation modifications of this protein may prove useful for the design of anti-HPV vaccines. (C) 1999 Elsevier Science B.V. All rights reserved.

A role for protein kinase B beta/Akt2 in insulin-stimulated GLUT4 translocation in adipocytes

Insulin stimulates glucose uptake into muscle and fat cells by promoting the translocation of glucose transporter 4 (GLUT4) to the cell surface. Phosphatidylinositide 3-kinase (PI3K) has been implicated in this process. However, the involvement of protein kinase B (PKB)/Akt, a downstream target of PI3K in regulation of GLUT4 translocation, has been controversial. Here we report that microinjection of a PKB substrate peptide or an antibody to PKB inhibited insulin-stimulated GLUT4 translocation to the plasma membrane by 66 or 56%, respectively. We further examined the activation of PKB isoforms following treatment of cells with insulin or platelet-derived growth factor (PDGF) and found that PKB beta is preferentially expressed in both rat and 3T3-L1 adipocytes, whereas PKB alpha expression is down-regulated in 3T3-L1 adipocytes. A switch in growth factor response was also observed when 3T3-L1 fibroblasts were differentiated into adipocytes. While PDGF was more efficacious than insulin in stimulating PKB phosphorylation in fibroblasts, PDGF did not stimulate PKB beta phosphorylation to any significant extent in adipocytes, as assessed by several methods. Moreover, insulin, but not PDGF, stimulated the translocation of PKB beta to the plasma membrane and high-density microsome fractions of 3T3-L1 adipocytes. These results support a role for PKB beta in insulin-stimulated glucose transport in adipocytes.

Wall material properties of yeast cells. Part II. Analysis

In the preceding paper (Part I) force-deformation data were measured with the compression experiment in conjunction with the initial radial stretch ratio and the initial wall-thickness to cell-radius ratio for baker's yeast (Saccharomyces cerevisiae). In this paper, these data have been analysed with the mechanical model of Smith et al. (Smith, Moxham & Middelberg (1998) Chemical Engineering Science, 53, 3913-3922) with the wall constitutive behaviour defined a priori as incompressible and linear-elastic. This analysis determined the mean Young's modulus ((E) over bar), mean maximum von Mises stress-at-failure (<(sigma)over bar>(VM,f)) and mean maximum von Mises strain-at failure (<(epsilon)over bar>(VM,f)) to be (E) over bar = 150 +/- 15 MPa, <(sigma)over bar>(VM,f) = 70 +/- 4 MPa and <(epsilon)over bar>(VM,f) = 0.75 +/- 0.08, respectively. The mean Young's modulus was not dependent (P greater than or equal to 0.05) on external osmotic pressure (0-0.8 MPa) nor compression rate (1.03-7.68 mu m/s) suggesting the incompressible linear-elastic relationship is representative of the actual cell-wall constitutive behaviour. Hydraulic conductivities were also determined and were comparable to other similar cell types (0-2.5 mu m/MPa s). The hydraulic conductivity distribution was not dependent on external osmotic pressure (0-0.8 MPa) nor compression rate (1.03-7.68 mu m/s) suggesting inclusion of cell-wall permeability in the mechanical model is justified. <(epsilon)over bar>(VM,f) was independent of cell diameter and to a first-approximation unaffected (P greater than or equal to 0.01) by external osmotic pressure and compression rate, thus providing a reasonable failure criterion. This criterion states that the cell-wall material will break when the strain exceeds <(epsilon)over bar>(VM,f) = 0.75 +/- 0.08. Variability in overall cell strength during compression was shown to be primarily due to biological variability in the maximum von Mises strain-at-failure. These data represent the first estimates of cell-wall material properties for yeast and the first fundamental analysis of cell-compression data. They are essential for describing cell-disruption at the fundamental level of fluid-cell interactions in general bioprocesses. They also provide valuable new measurements for yeast-cell physiologists. (C) 2000 Elsevier Science Ltd. All rights reserved.

Lorneamides A and B: Two new aromatic amides from a southern Australian marine actinomycete

A marine actinomycete (MST-MA190) isolated from a sample of beach sand collected near Lorne on the southwest coast of Victoria, Australia, has yielded two new aromatic amides, lorneamide A (1) and lorneamide B (2). The lorneamides belong to a novel class of tri-alkyl-substituted benzenes, and their structures were determined by spectroscopic methods.

Characterization of peroxisome proliferator-activated receptor alpha in normal rat mammary gland and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced mammary gland tumors from rats fed high and low fat diets

Normal Sprague-Dau ley rat mammary gland epithelial cells and mammary gland carcinomas induced by 2-amino-1 -methyl-6-phenylimidazo[4,5-b]pyridine, a carcinogen found in the diet, were examined for the expression of peroxisome proliferator-activated receptor alpha (PPAR alpha). PPAR alpha mRNA and protein was detected in normal and tumor tissue by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. By quantitative RT-PCR, carcinomas had a 12-fold higher expression than control mammary glands, a statistically significant difference. PPAR alpha expression was examined in carcinomas and normal tissues from rats on high fat (23.5/% corn oil) and low fat (5% corn oil) diets. Although neither carcinomas, nor control tissues showed statistically significant differences between the two diet groups, PPAR alpha expression was the highest in carcinomas from rats on the high fat diet. The expression of PPAR alpha in normal mammary gland and its significant elevation in mammary gland carcinomas raises the possibility of its involvement in mammary gland physiology and pathophysiology. (C) 2000 Published by Elsevier Science Ireland Ltd. All rights reserved.

Phorbasin B and C: Novel diterpenes from a southern Australian marine sponge, Phorbas species

A southern Australian Phorbas sp. has yielded the novel diterpenes phorbasin B (2) and phorbasin C (3). Phorbasins B and C possess a hitherto unknown carbon skeleton, and their structures were assigned on the basis of detailed spectroscopic analyses.

Energy and averages in the mechanics of granular materials

We derive a general thermo-mechanical theory for particulate materials consisting of granules of arbitrary whose material points possess three translational and three independent rotational degrees of freedom. Additional field variables are the translational and rotational granular temperatures, the kinetic energies shape and size. The kinematics of granulate is described within the framework of a polar continuum theory of the velocity and spin fluctuations respectively and the usual thermodynamic temperature. We distinguish between averages over particle categories (averages in mass/velocity and moment of inertia/spin space, respectively) and particle phases where the average extends over distinct subsets of particle categories (multi phase flows). The relationship between the thermal energy in the granular system and phonon energy in a molecular system is briefly discussed in the main body of the paper and discussed in detail in the Appendix A. (C) 2001 Elsevier Science B.V. All rights reserved.

On the minimality of the p-harmonic map - x/vertical bar x vertical bar : B-n -> Sn-1

We prove that for any real number p with 1 p less than or equal to n - 1, the map x/\x\ : B-n --> Sn-1 is the unique minimizer of the p-energy functional integral(Bn) \delu\(p) dx among all maps in W-1,W-p (B-n, Sn-1) with boundary value x on phiB(n).

Finger and A-B ridge count, and fluctuating asymmetry in psychosis: A catchment area case-control study

Dermatoglyphic measures are of interest to schizophrenia research because they serve as persistent markers of deviant development in foetal life. Several studies have reported alterations in A–B ridge counts, total finger ridge counts and measures related to asymmetry in schizophrenia. The aim of this study was to assess these measures in an Australian catchment area, case-control study. Individuals with psychosisŽns246.were drawn from a catchment-area prevalence study, and well controlsŽns229. were recruited from the same area. Finger and palm prints were taken usingan inkless technique and all dermatoglyphic measures were assessed by a trained rater blind to case status. The dermatoglyphic measures Žfinger ridge count, A–B ridge count, and their derived asymmetry measures. were divided into quartiles based on the distribution of these variables in controls. The main analysis Žlogistic regression controlled for age and sex.examined all psychotic disorders, with planned subgroup analyses comparing controls with Ž1. nonaffective psychosis Žschizophrenia, delusional disorder, schizophreniform psychosis, atypical psychosis.andŽ2. affective psychosis Ždepression with psychosis, bipolar disorder, schizoaffective psychosis.. There were no statistically significant alterations in the odds of havinga psychotic disorder for any of the dermatoglyphic measures. The results did not change when we examined affective and nonaffective psychosis separately. The dermatoglyphic features that distinguish schizophreniar psychosis in other studies were not identified in this Australian study. Regional variations in these findings may provide clues to differential ethnicrgenetic and environmental factors that are associated with schizophrenia. The Stanley Foundation supported this project.

Phoriospongin A and B: Two new nematocidal depsipeptides from the Australian marine sponges Phoriospongia sp and Callyspongia bilamellata

Bioassay-directed fractionation of two southern Australian sponges, Phoriospongia sp. and Callyspongia bilamellata, yielded two new nematocidal depsipeptides, identified as phoriospongins A (1) and B (2). The structures of the phoriospongins were determined by detailed spectroscopic analysis and comparison with the previously reported sponge depsipeptide cyclolithistide A (3), as well as ESIMS and HPLC analysis of acid hydrolysates. It is noteworthy that the unique and yet structurally related metabolites 1-3 are found in sponges spanning three taxonomic orders, Poescilosclerida, Haplosclerida, and Lithistida.

CD40 ligation conditions dendritic cell antigen-presenting function through sustained activation of NF-kappa B

An understanding of the biochemical control of dendritic cell (DC) differentiation/activation is essential for improving T cell immunity by various immunotherapeutic approaches, including DC immunization. Ligation of CD40 enhances DC function, including conditioning for CTL priming. NF-kappaB, and particularly RelB, is an essential control pathway for myeloid DC differentiation. Furthermore, RelB regulates B cell Ag-presenting function. We hypothesized that CD40 ligand (CD40L) and TNF-alpha, which differ in their capacity to condition DC, would also differ in their capacity to activate NF-kappaB. DC differentiated for 2 days from monocytes in the presence of GM-CSF and IL-4 were used as a model, as NF-kappaB activity was constitutively low. The capacity of DC to activate T cells following CD40L treatment was enhanced compared with TNF-alpha treatment, and this was NF-kappaB dependent. Whereas RelB/p50 translocation induced by TNF-alpha was attenuated after 6 h, RelB/p50 nuclear translocation induced by CD40L was sustained for at least 24 h. The mechanism of this difference related to enhanced degradation of IkappaBalpha following CD40L stimulation. However, NF-kappaB activation induced by TNF-alpha could be sustained by blocking autocrine IL-10. These data indicate that NF-kappaB activation is essential for T cell activation by DC, and that this function is enhanced if DC NF-kappaB activation is prolonged. Because IL-10 moderates DC NF-kappaB activation by TNF-alpha, sustained NF-kappaB activation can be achieved by blocking IL-10 in the presence of stimuli that induce TNF-alpha.

Semisolid casting of AlSi7Mg0.35 alloy produced by low-temperature pouring

AlSi7Mg0.35 alloy was cast into permanent moulds using different pouring temperatures (725 to 625degreesC). As the pouring temperature decreased, the as-cast microstructure changed from a coarse dendritic structure, through fine equiaxed grains to fine rosette-like grains. The as-cast materials were then partially remelted and isothermally held at 580degreesC prior to semisolid casting into a stepped die. The feedstock material cast from a high temperature filled only half the die, with severe segregation and other defects. The low-temperature-poured material completely filled the die with negligible porosity. The quality of semisolid castings is significantly affected by the microstructure of the semisolid feedstock material that arises from a combination of as-cast and subsequent thermal treatment conditions. The paper describes (a) the influence of pouring temperature on the microstructure of feedstock; (b) microstructure evolution through remelting and (c) the quality of semisolid castings produced with this material. For A17Si0.35Mg alloy, low temperature pouring in the range of 625-650degreesC followed by suitable isothermal holding treatment can result in good quality semisolid casting.