Molecular evidence for the expression of nicotinic acetylcholine receptor alpha-chain in mouse thymus.

The presence and structure of nicotinic acetylcholine receptor (nAChR) in the thymus has been a subject of interest for many years because of its possible role in the pathogenesis of the autoimmune disease myasthenia gravis. Using the polymerase chain reaction with primers specific for the alpha-chain of nAChR (nAChR-alpha), an 880-bp homologous band was found after amplification of cDNA prepared from mouse thymus, thymic medullary and cortical epithelial cell lines, but not from thymocytes or kidney. Sequencing of the polymerase chain reaction product from the thymus and thymic medullary and cortical epithelial lines showed identity with skeletal muscle nAChR-alpha over the region examined. This region includes the domains of the molecule on which B cell and T cell autoantigenic targets have been described. No evidence was found in mouse tissue for the exon 3A, which has been described in human muscle and the human rhabdomyosarcoma cell line TE671. Our results provide evidence at the RNA level for the expression of the nAChR-alpha on stromal cells but not on thymocytes in normal murine thymus and are consistent with a role for intrathymic autoantigen expression in the pathogenesis of myasthenia gravis.     

Identification of N5-methyl-N5-formyl-2,5,6-triamino-4-hydroxypyrimidine as a major adduct in rat liver DNA after treatment with the carcinogens, N,N-dimethylnitrosamine or 1,2-dimethylhydrazine

Human Tamm-Horsfall urinary glycoprotein from an individual of the blood group Sd(a+) phenotype was tritium-labelled by treatment with galactose oxidase and sodium boro[3H]hydride and was then digested with endo-beta-galactosidase. A series of dialysable, labelled fragments was released from which a pentasaccharide was isolated that strongly inhibited the agglutination of Sd(a+) red cells by human anti-Sda serum and hence contained the Sda determinant structure. Reduction, methylation analysis and sequential exo-glycosidase digestion established the structure of the pentasaccharide as: GalNAc beta(1 leads to 4)[NeuAc(2 leads to 3)]Gal beta(1 leads to 4)GlcNAc beta(1 leads to 3)Gal     

Restriction fragment length polymorphisms of horse class II MHC genes observed using various human alpha-and beta-chain cDNA probes

Genomic DNA isolated from 20 horses was digested with up to six restriction endonucleases and subjected to southern blot hybridization analysis using various human class II alpha- and beta-chain cDNA probes. A high degree of restriction fragment length polymorphism (RFLP) was found for the DQ alpha, DP beta, DQ beta and DR beta probes, about 20 polymorphic bands being detected for each. DR alpha showed 2-4 polymorphic bands, whereas no evidence for DP alpha-like genes was found. A number of correlations of RFLPs with individual alloantisera were apparent.     

Molecular heterogeneity of D-end products detected by anti-H-2.28 sera

Immunoprecipitation of NP-40 lysates of 125I-labeled lymph-node cells with different anti-H-2 sera and with anti-Qa-2 serum has shown that the BALB/cByA strain (H-2d, Qa-2-negative) expresses, besides H-2Ld, another molecule that is not detectable in the BALB/c-H-2dm2 strain. Electrophoresis in SDS polyacrylamide gels indicated that this molecule, provisionally designated Lq, has an apparent molecular weight of 41000 daltons, in contrast to approximately 49000 daltons for H-2Kd and H-2Ld, and 47000 daltons for H-2Dd molecules. The anti-Qa-2 serum precipitated from the Qa-2-positive strains BALB/cHeA but not from the Qa-2-negative strains BALB/cByA and BALB/c-H-2dm2 a protein that gave a very strong band corresponding to the molecular weight 41000 daltons in the gel electrophoresis. The biochemical characteristics of the Lq molecule are thus more similar to those of Qa-2 than of H-2 antigens.     

Surface Plasmodium sugar-binding components evidenced by fluorescent neoglycoproteins

A sugar-binding component was shown to be present at the surface of endoerythrocytic Plasmodium berghei and P. chabaudi stages and merozoites by means of a panel of neoglycoproteins using fluorescence microscopy and flow cytofluorometry. The protein nature of this material was ascertained by the loss of sugar-binding capacity upon trypsin treatment. This lectin-like protein primarily bound neoglycoprotein-borne N-acetylglucosamine and secondarily bound neoglycoprotein-borne alpha-D-glucose, beta-D-galactose and alpha-L-fucose. These results suggest the presence of at least one type of lectin-like protein with an extended binding site accomodating several sugar units, and define its localization on the parasite surface.     

Comparative greenhouse study of Eucalyptus grandis in vitro plantlets and half-sib seedlings,II. Dry matter accumulation and relative distribution

In vitro directly micropropagated plantlets from three selected five-year-old Eucalyptus grandis Hill ex. Maiden hybrids were compared to their related half-sib seedlings for growth and growth pattern parameters under greenhouse conditions used for operational seedling production. The oven dry weights were determined from stem, leaf, and root samples collected every 40 days for four times. Relative growth rate, net assimilation rates and shoot:root ratio were calculated. Survival was 98% and 95% for plantlets and seedlings, respectively. Significant differences were observed between parents in terms of shoot and root dry weights and their ratios with similar ranking among plantlets and seedlings, suggesting genetic control over these traits. Plantlets started with significantly higher root: shoot ratios and stem, leaf, root, and total dry weight. Although seedlings had higher relative growth and net assimilation rates, all the initial differences decreased sharply over time.     

Identification of bacteria of dairy origin using miniaturized test-systems

One hundred and forty-four isolates of dairy origin were identified using a number of commercially available test kits. Agreement between these kits was acceptable for assignment of isolates to genera, but there was some variation between the kits for the identification of the isolates at specific level. The reasons for these variations are discussed. In terms of cost, ease of performance and accuracy, the identification systems manufactured by Roche Products Ltd (viz. the Oxi/Ferm tube and the Enterotube) are considered to be the best suited to the routine identification of dairy micro-organisms.     

The histology and calcification of regenerating scales in the blackspotted topminnow, Fundulus olivaceus (Storer)

The regenerating scale and tissues comprising the scale pocket of Fundulus olivaceus were examined microscopically at specific intervals. Scale removal resulted in a thickening of the epidermis which persisted through the early stages of regeneration. This thickening was due in part to the appearance of columnar basal cells which divided producing cells that became mucous cells and squamous cells. The scale regenerated as a relatively large plate of bone which first appeared between layers of scleroblasts on the floor of the scale pocket and then grew producing circuli and radii. By the fourth day of regeneration, calcium was observed in the cytoplasm of the scleroblasts and at randomly distributed foci in the osseous portion of the scale. The osseous layer was completely calcified by 15 days of regeneration.