Metabolism of the mutagen MeIQx in vivo: metabolite screening by liquid chromatography-thermospray mass spectrometry

2-Amino-3,8-dimethylimidazo(4,5-f)quinoxaline (MeIQx) is a potent mutagen found in cooked food. MeIQx and its isotopically labelled (13C, 15N2 and 14C) analogues were synthesised and used for metabolic studies in vivo. An equimolar mixture of MeIQx and its 13C, 15N2 stable isotope labelled analogue (containing tracer amounts of 14C-MeIQx) was given intraperitoneally to mice. Some 67% of the radioactivity was eliminated in urine and faeces within 24h. Four radiolabelled species were observed when urine was analysed by HPLC, corresponding to unchanged MeIQx and three more polar metabolites. Urine was analysed directly by HPLC-thermospray mass spectrometry. Four signals were observed containing the characteristic 1:1 isotopic doublet, corresponding to unchanged MeIQx, an MeIQx glucuronide, and two uncharacterized metabolites.     

The evolution of human chromosome 21: evidence from in situ hybridization in marsupials and a monotreme.

We have mapped five human chromosome 21 (HSA 21) markers in marsupials and a monotreme, two major groups of mammals that diverged from eutherians 130-150 and 150-170 million years before present (MYrBP), respectively. We have found that these genes map to two distinct autosomal sites, one containing SOD1/CBR/BCEI and the other containing ETS2/INFAR, in the marsupials Macropus eugenii and Sminthopsis macroura (which belong to orders that diverged 40-80 MYrBP), as well as in the monotreme Ornithorhynchus anatinus (the platypus). Since marsupials and monotremes diverged independently from eutherians, these data suggest that HSA 21 genes were originally located in two separate autosomal blocks. In another Sminthopsis species, SOD1 is linked to TRF (a marker on HSA 3q), suggesting that the ancestral SOD1/CBR/BCEI region also included HSA 3 markers. We suggest that these blocks became fused early in the eutherian evolution to form a HSA 3/21 chromosome, which has remained intact in artiodactyls, but has been independently disrupted in both the primate and rodent lineages.     

Immunoregulation in disseminated murine histoplasmosis: disturbances in the production of interleukins 1 and 2

Production of IL 1 and IL 2 by splenocytes from C57BL/6 mice was measured at wk 1, 3, 8, and 14 after i.v. inoculation with 6 X 10(5) Histoplasma capsulatum (Hc) yeasts. As compared with age-matched controls, IL 1 production by splenocytes from Hc-infected mice was reduced severely at wk 1 and 3 of infection, greater than normal at wk 8, and within normal range at wk 14. IL 2 production was also reduced at wk 1 and 3 of infection; it was normal at wk 8 and was elevated at wk 14. Indomethacin and catalase failed to restore IL 1 production by splenocytes from infected mice, and exogenous IL 1 did not augment IL 2 production by these cells. A factor capable of suppressing the activity of IL 2 was detected in supernatants of concanavalin A-stimulated splenocytes from infected animals at wk 1 and 3 of infection, respectively. No factor capable of suppressing IL 1 activity was detected. Thus, the deficits of cell-mediated immunity in mice with systemic Hc infection may derive, in part, from impaired amplification of the immune response consequent to abnormal generation of IL 1 and IL 2.     

Nitrospira marina gen. nov. sp. nov.: a chemolithotrophic nitrite-oxidizing bacterium

A new chemolithotrophic nitrite-oxidizing bacterium, for which the name Nitrospira marina is proposed, was isolated from the Gulf of Maine. N. marina is a Gramnegative curved rod which may form spirals with 1 to 12 turns. Cells have a unique periplasmic space and lack intracytoplasmic membranes and carboxysomes. N. marina is an obligate chemolithotroph, but best growth is obtained in a mixotrophic medium. N. marina may be one of the most prevalent nitrite-oxidizing bacteria in some oceanic environments. Type strain is field with American Type Culture Collection (ATCC 43039).     

Use of 6-diazo-5-oxo-l-norleucine to study interaction between myocardial glycoconjugate secretion and endothelial activation in the early embryonic chick heart

The glutamine analog, 6-diazo-5-oxo-l-norleucine (DON), a glycoconjugate inhibitor, was used to probe the relationships between myocardial secretion of extracellular matrix and endothelial differentiation and formation of cushion mesenchyme (primordia of AV values). When DON was given to stage 12 chick embryos maintained in shell-less culture, the myocardial secretion gradient of glucose- and sulfate-labeled matrix was blocked. Concomitantly, the endothelium failed to complete activation but continued to divide and incorporate thymidine. By varying DON concentration, two distinct phases of endothelial differentiation were identified: the first (labile to 0.5 μg) involved hypertrophy, the second (labile to 0.25 μg) acquisition of migratory appendages with resultant mesenchyme formation. Glucosamine + DON (but not inosine, glucose, or glutamine) restored the matrical secretion gradient and to varying degrees both phases of endothelial activation. Endothelia totally suppressed from forming mesenchyme in situ acquired this capacity when explanted into three-dimensional collagen gel culture. The capacity was enhanced by glucosamine given in situ as an inhibitory override, dependent upon serum concentration, inhibited by heat-inactivated serum or by adding DON to the medium, but unaffected by hyaluronate. These results were compared to those obtained by co-culturing endothelium and myocardium and discussed in terms of the hypothesis that cushion mesenchyme formation results from an epithelial interaction mediated by glycoconjugates.     

The action of nitrous acid on C-teichoic acid (C-substance) from the walls of Diplococcus pneumoniae

1. C-teichoic acid (C-substance) from the walls of Diplococcus pneumoniae contained free amino groups accessible to attack by nitrous acid. Treatment with nitrous acid, followed by reduction with borohydride and hydrolysis with acid, gave ribitol, glucitol and their respective phosphates. 2. Hydrolysis of the polymer with alkali followed by treatment of products with nitrous acid yielded glucose. 3. When alkali hydrolysis was followed by treatment with a phosphomonoesterase, nitrous acid degradation of C-substance yielded glucose and a disaccharide identified as 2-O-(N-acetylgalactosaminyl)-d-ribitol. 4. A partial structure for C-teichoic acid was deduced in which the order of the constituent residues and the position of phosphodiester linkages were established.