Differential mediator production by dendritic cells upon toll-like receptor stimulation does not impact T cell cytokine expression.

Dendritic cells are key components of successful immunological responses bridging innate and adaptive defenses. In this study we wanted to know whether ligation of toll-like receptors (TLR) expressed by dendritic cells would induce differential proinflammatory mediator expression and whether these dendritic cells would differentially impact T cell function. For this purpose bone marrow-derived dendritic cells from OTII mice were used. The dendritic cells showed detectable levels of TLR1, 2, 4, 6, 7, 8 and 9, with TLR2 and TLR4 expressed at the highest levels. To determine whether TLR ligation differentially influenced proinflammatory mediator expression the dendritic cells were stimulated with peptidoglycan (PGN) or lipopolysaccharide (LPS) for TLR2 or TLR4, respectively. Comparisons were made to dendritic cells exposed to TNF-alpha or saline as controls. Whereas, both LPS and PGN were equally effective at inducing CXCL1 and TNF-alpha expression from the dendritic cells, LPS was unique at inducing CCL2 expression, and PGN was unique at inducing IL-1beta expression. Despite these differences, LPS and PGN treated dendritic cells were equally effective at eliciting IFN-gamma expression from T cells in an antigen-specific manner. These data indicate that ligation of TLR by components of Gram+ and Gram- bacteria differentially influence dendritic cell proinflammatory mediator expression, and that differential mediator production by dendritic cells upon TLR stimulation does not impact T cell cytokine production.

Updating small area population estimates in England and Wales.

"Population estimates have important implications for resource allocation within government and commerce, and are often assumed to be without error. Currently, central government provides annual population estimates for all the local and health authority districts in Britain, but estimates are needed for smaller areas, typically for electoral wards and postal sectors. Small area estimates are provided by some local authorities and commercial organizations, using different methods; the accuracy of these estimates is modelled here within a multilevel framework. Certain characteristics of the small area and of the method of estimation are included as explanatory variables. Results show that the method of estimation used is of great importance."

Constructing an urban deprivation index: a way of meeting the need for flexibility.

The authors examine the use of census data to construct an urban deprivation index for the United Kingdom, with special attention given to the need for flexibility. "A single index is rejected in favour of a matrix of results which captures the complex geography of deprivation. The matrix of districts includes measures of the degree of deprivation, its spatial extent, its intensity, and the spatial distribution of deprivation at the enumeration district scale. The profiles of various districts are discussed to illustrate the use of the matrix."

Modification of hemoglobin - tetrameric stabilization.

Advantage has been taken of the unique affinity of DBBF for hemoglobin to stabilize the T state and crosslink tetrameric hemoglobin. The oxygen affinity has been further decreased by using PLP to produce a chemically unique molecule with a P50 of 32 under physiologic conditions. Furthermore, the modification is specific, requires only reagents that are commercially available or easily synthesized, and can be prepared in large quantities with up to 80% yield. The unique modified hemoglobin was purified by HPLC and the crosslink was found between the beta chains. This derivative, pyridoxalated-fumarate hemoglobin, sustained life in five rats that were 95% exchanged-transfused with the solution. Preliminary in vivo tests support this derivative as a desirable oxygen-transporting resuscitation fluid.

The preparation of lipid-free extracts of drugs from blood for gas chromatographic analysis.

A method is presented to separate co-extracted biological materials, in particular lipids, from drugs in chloroform extracts of blood. The chloroform is evaporated and the residue is dissolved in a small volume of petroleum ether and partitioned with eight times this volume of 5% aqueous hydrochloric acid. The acidic, basic, and neutral drugs are extracted into the aqueous phase, leaving unwanted materials in the petroleum ether. The recoveries of drugs representative of a variety of types are presented. Reduction of the levels of lipids by factors on the order of 1000-fold is reported.

Fungal growth on C1 compounds: quantitative aspects of growth of a methanol-utilizing strain of Trichoderma lignorum in batch culture.

A study was made of some salient parameters that influence growth of the methanol-utilizing fungus Trichoderma lignorum growing in batch culture on a minimal medium containing methanol as the sole source of carbon. Maximum cell yield was recorded at the expense of 1.58 g of methanol per liter. Inhibition was observed with methanol concentrations in excess of 4.7 g/liter. The optimum temperature for fungal growth was 23 degrees C. Growth of the fungus was directly proportional to an inorganic nitrogen concentration up to 0.2 g of NH4NO3 per liter. No inhibition of growth occurred at any concentration of NH4NO3 up to 11 g/liter. The pH of the growth medium decreased from 7.0 to 3.5 during growth of the fungus on methanol, which may have been due, in part, to the accumulation of trace amounts of organic acids in the growth medium. An analysis of the commercial potential of the fungus, as a source of edible protein, indicated that the strain of methanol-utilizing T. lignorum used was uneconomical in terms of the yield and the specific growth rate.

Inhibition of DNA polymerase from L1210 murine leukemia by a sulfhydryl reagent from agaricus bisporus.

The 490 quinone, a natural sulfhydryl-arylating reagent from the mushroom, Agaricus bisporus, markedly inhibited L1210 murine leukemia DNA polymerase alpha while resulting in little inhibition of DNA polymerase beta from this source. This quinone was more strongly inhibitory than p-chloromercuri-benzoate or N-ethylmaleimide and was less readily neutralized by sulfhydryl-containing molecules such as dithioerythritol. Preliminary experiments indicate that DNA protects DNA polymerase alpha from inhibition by the 490 quinone. The inhibition of DNA synthesis by quinone 490 may contribute significantly to the cytotoxicity of this compound and to the potential of gamma-L-glutaminyl-4-hydroxybenzene as an antitumor agent.

Isolation and characterization of two glycoproteins from patients with alveolar proteinosis.

The saline-insoluble particulate material obtained by pulmonary lavage from four patients with alveolar proteinosis was analyzed for total lipid (52%), protein (44%), and carbohydrate (4%). Sodium dodecyl sulfate-gel electrophoresis revealed only three major peptides in these patients. Molecular weights (from the gels) were 69,000, 62,000, and 36,000, and the latter two peptides were periodic acid-Schiff stain-positive. Amino-acid and sugar analyses were performed on the peptides cut from the gels and on peptides purified by Sephadex G-200 chromatography. The 69,000-molecular-weight peptide contained no carbohydrate. Antibody studies and aminoacid analysis indicated that it was albumin. The 62,000-molecular-weight peptide contained 1% hydroxyproline, 1% hydroxylysine, 10% glycine, and 9% carbohydrate. The 36,000-molecular-weight peptide contained 1.2% hydroxyproline, 1% hydroxylysine, 13% glycine, 1.4% sialic acid, 2.6% glucose, 2.4% galactose, 2% mannose, 0.8% fucose, 0.4% glycosamine, and 0.6% galactosamine. Proteins extracted from kidney glomeruli with similar amino-acid and carbohydrate composition have been observed previously.

Experimental carcinogenesis of the lung. II. Influence of phenols in the production of carcinoma.

The acidic Fractions (including phenols) were separated from two somewhat different coal tars. Various blends and an original tar produced aerosols to which 5 groups of male C3H/HeJ mice were exposed 2 hours, 3 times weekly, for 55 weeks. Animals were killed at intervals. The lungs and tracheas of all mice were examined grossly and microscopically for neoplasms or relevant morphologic changes. After 46 weeks, 32 survivors in groups 2 and 4, which received similar aerosols containing phenols, had 4 incidences of adenocarcinoma, 19 of intrabronchial adenoma, and 10 of squamous metaplasia. In 20 survivors of group 3, which received the same far without phenols, there was no incidence of adenocarcinoma, 11 had intrabronchial adenoma and 2 had squamous metaplasia.