Human peripheral blood monuclear cells transfected with messenger RNA stimulate antigen-specific cytotoxic T-lymphocytes in vitro

The efficiency of test vaccines needs to be evaluated by quantification of the triggered cellular immune response. Usually, for these assays, autologous target cells expressing the vaccine antigen are required. In the context of messenger RNA (mRNA)-based vaccinations, the target cells used for the read-out are mRNA-transfected monocyte-derived dendritic cells (Mo-DCs). Their production typically requires samples of 100 ml blood from the patients, and limits the number of assays that can be performed. We show here that fresh peripheral blood mononuclear cells (PBMCs) can be transfected with mRNA by electroporation. Such cells are as efficient as mRNA-transfected Mo-DCs for their ability to activate memory T cells in vitro. Thus, mRNA-transfected PBMCs are a convenient replacement of mRNA-transfected Mo-DCs for the in vitro monitoring of natural or vaccine-induced immune responses.Received 17 February 2005; received after revision 1 May 2005; accepted 7 Juni 2005     

Creating high‐frequency national accounts with state‐space modelling: a Monte Carlo experiment

This paper assesses a new technique for producing high‐frequency data from lower frequency measurements subject to the full set of identities within the data all holding. The technique is assessed through a set of Monte Carlo experiments. The example used here is gross domestic product (GDP) which is observed at quarterly intervals in the United States and it is a flow economic variable rather than a stock. The problem of constructing an unobserved monthly GDP variable can be handled using state space modelling. The solution of the problem lies in finding a suitable state space representation. A Monte Carlo experiment is conducted to illustrate this concept and to identify which variant of the model gives the best monthly estimates. The results demonstrate that the more simple models do almost as well as more complex ones and hence there may be little gain in return for the extra work of using a complex model. Copyright © 2001 John Wiley & Sons, Ltd.     

Light on the structure of thromboxane A2 receptor heterodimers

The structure-based design of a mutant form of the thromboxane A(2) prostanoid receptor (TP) was instrumental in characterizing the structural determinants of the hetero-dimerization process of this G protein coupled receptor (GPCR). The results suggest that the hetero-dimeric complexes between the TPα and β isoforms are characterized by contacts between hydrophobic residues in helix 1 from both monomers. Functional characterization confirms that TPα-TPβ hetero-dimerization serves to regulate TPα function through agonist-induced internalization, with important implications in cardiovascular homeostasis. The integrated approach employed in this study can be adopted to gain structural and functional insights into the dimerization/oligomerization process of all GPCRs for which the structural model of the monomer can be achieved at reasonable atomic resolution.     

Conformational studies ofcis-10-methyl-2-decalones

Zusammenfassung Im Kernresonanzspektrum einiger substituiertercis-2-Decalone wurden die chemischen Verschiebungen angulärer Methylgruppen untersucht. Dabei wurde festgestellt, dass die Verbindungen vorwiegend in der nicht-steroidalen Sessel-Sessel-Konformation vorliegen. Das Vorzeichen der ORD Cotton-Kurven dieser Ketone muss durch das untergeordnete Vorhandensein einer Konformation sehr starker Amplitude, wie z. B. der Twist-Form, bedingt sein.     

Production and detection of antibody against estradiol receptor in the calf uterus. Interaction with the estrogen receptor from the hen oviduct]

The antibodies against estrogen receptor were obtained after injecting Rabbits with a cytoplasmic receptor fraction isolated from Calf uterus. The estrogen receptor was partially proteolysed by the action of trypsin and subsequently purified by affinity chromatography (purification 4,000 to 10,000 fold, to a purity of 5-20%). The affinity of the antibody for the proteolysed receptor is KD approximately 1 nM and serum titres have reached values of approximately 50 nM. The values remained constant after the third injection. Preliminary results indicate that the antibody has approximately the same affinity for "native" cytoplasmic estrogen receptor from Calf uterus, as well as for the "trypsinized" forms of estrogen receptor isolated from Calf uterine cytosol and Hen oviduct nuclei.     

Levamisole inhibits mineral mobilisation, lactate production and lysosomal enzyme release from cultured bones.

Levamisole added to cultured calvarial bones inhibited spontaneous bone resorption, as indicated by reduced release of calcium and inorganic phosphate to the medium. In addition, levamisole reduced lactate production and release of lysosomal enzymes.     

Peroxidase activity and thiocyanate accumulation in salivary glands

Summary Salivary glands with high, low, or no peroxidase activity do not differ in [S¹⁴CN^–] after the i.v. injection of KS¹⁴CN, nor do the glands differ from blood and muscle in [S¹⁴CN^–]. The content of SCN^– in a salivary gland does not mirror the gland's participation in the peroxidase-mediated antimicrobial mechanism.     

Differences in cardiac myosin light chain LC1 among human, monkey and sheep

The atrial and ventricular myosin light chains of human, monkey and sheep hearts were compared by dodecylsulfate polyacrylamide gel electrophoresis. The atrial light chain 2 and ventricular light chain 2 are similar among these mammals. However, the atrial light chain 1 of monkey has different electrophoretic mobility from those of human and sheep. The monkey ventricular light chain 1 has same mobility as that of sheep but different from that of human.