The chromosomes ofAzima tetracantha (Salvadoraceae)

Azima tetracantha has an asymmetrical karyotype with large chromosomes and a large amount of heterochromatin. The haploid number (n = 11) may represent the base number of the family. However, a possible secondary origin of this base number is also considered.     

Suppression of experimental autoimmune encephalomyelitis by oral administration of myelin basic protein. II. Suppression of disease and in vitro immune responses is mediated by antigen-specific CD8+ T lymphocytes

We have previously demonstrated that the oral administration of guinea pig myelin basic protein (MBP) protects Lewis rats against the induction of experimental autoimmune encephalomyelitis (EAE) when subsequently immunized with guinea pig MBP in CFA. In addition, animals made orally tolerant to MBP also have diminished proliferative and antibody responses to MBP, but not to other Ag. Nonetheless, the mechanism of oral tolerance to MBP in the EAE model remains undefined. In the present study, we report that T cells isolated from the spleen and mesenteric lymph nodes of MBP orally tolerized animals can adoptively transfer protection against EAE. Furthermore, these T cells are of the CD8+ subclass. In addition, CD8+ T cells from MBP orally tolerized animals also suppress in vitro proliferative responses and antibody responses to MBP in an Ag-specific fashion. These results demonstrate that active cellular mechanisms are initiated after oral administration of an autoantigen that can down-regulate an experimental autoimmune disease and provide the basis for the isolation and characterization of the cells mediating both in vivo and in vitro suppression.     

Folylpolyglutamate Derivatives of Pisum sativum L. Determination of Polyglutamate Chain Lengths by High Performance Liquid Chromatography Following Conversion to p-aminobenzoylpolyglutamates

The folypolyglutamate derivatives of pea seedlings (Pisum sativumL. cv. Homesteader) were extracted in the presence of 2-mercaptoethanoland cleaved to p-aminobenzoylpolyglutamates by treatment withZn-HCl. Azo dyes were formed by reaction with naphthylethylenediamine and purified by polyacrylamide gel chromatography. p-Aminobenzoylpolyglutamateswere regenerated from these dyes by Zn treatment and then concentratedin vacuo. These derivatives were separated according to glutamylchain length by high performance liquid chromatography on WhatmanPartisil SAX columns. The folylpolyglutamates of 4 day old peacotyledons, pea leaves and isolated chloroplasts were mainlytetra- and pentaglutamates. These and folates of shorter chainlength were labelled when seeds and aerial shoots were incubatedwith p-aminobenzoate-[¹⁴C]. Labelling of the pentaglutamatewas reduced in seeds that were imbibed in the presence of 0.1mM methotrexate. Studies of cotyledon folylpolyglutamate synthetaseshowed that polyglutamate chain length was affected by incubationtime and the concentration of tetra-hydrofolate monoglutamatein the reaction system. ¹Present address: Department of Biology, University of Lethbridge,Alberta, Canada T1K 3M4 ²Present address: Department of Horticulture, Xiong-yue AgriculturalCollege, Xiong-yue, Liaoning Province, China (Received August 4, 1989; Accepted December 5, 1989)     

Isolation of peroxisomes from frozen human liver samples.

This paper shows the successful isolation of peroxisomes from human liver samples that were kept frozen at -70 degrees C. Purification of these peroxisomes was obtained by a combination of two subcellular fractionation techniques: differential centrifugation and isopycnic fractionation in Nycodenz density gradients. Peroxisome integrity was evaluated by latency measurements and by ultrastructural observation. The procedure described here may be useful for the isolation of other subcellular organelles from frozen human samples.     

Influence of benzo[a]pyrene diol epoxide chirality on solution conformations of DNA covalent adducts: the (-)-trans-anti-[BP]G.C adduct structure and comparison with the (+)-trans-anti-[BP]G.C enantiomer.

Benzo[a]pyrene (BP) is an environmental genotoxin, which, following metabolic activation to 7,8-diol 9,10-epoxide (BPDE) derivatives, forms covalent adducts with cellular DNA. A major fraction of adducts are derived from the binding of N2 of guanine to the C10 position of BPDE. The mutagenic and carcinogenic potentials of these adducts are strongly dependent on the chirality at the four asymmetric benzylic carbon atoms. We report below on the combined NMR-energy minimization refinement characterization of the solution conformation of (-)-trans-anti-[BP]G positioned opposite C and flanked by G.C base pairs in the d(C1-C2-A3-T4-C5-[BP]G6-C7-T8-A9-C10-C11).d(G12-G13-T14++ +-A15-G16-C17- G18-A19-T20-G21-G22) duplex. Two-dimensional NMR techniques were applied to assign the exchangeable and non-exchangeable protons of the benzo[a]pyrenyl moiety and the nucleic acid in the modified duplex. These results establish Watson-Crick base pair alignment at the [BP]G6.C17 modification site, as well as the flanking C5.G18 and C7.G16 pairs within a regular right-handed helix. The solution structure of the (-)-trans-anti-[BP]G.C 11-mer duplex has been determined by incorporating intramolecular and intermolecular proton-proton distances defined by lower and upper bounds deduced from NOE buildup curves as constraints in energy minimization computations. The BP ring spans both strands of the duplex in the minor groove and is directed toward the 3'-end of the modified strand in the refined structure. One face of the BP ring of [BP]G6 stacks over the C17 residue across from it on the partner strand while the other face is exposed to solvent.(ABSTRACT TRUNCATED AT 250 WORDS)     

The surface free energy of Leishmania mexicana amazonensis.

Surface charge of Leishmania mexicana amazonensis was investigated by direct zeta-potential determination and ultrastructural cytochemistry, and its surface tension was studied by measurements of the advancing contact angle formed by the parasite monolayers with drops of liquids of different polarities. Both virulent and avirulent promastigotes exhibited negatively charged surfaces with a zeta-potential of about -15 mV. Treatment of these cells with trypsin, alkaline phosphatase, or phospholipase C rendered their surfaces less negatively charged, whereas neuraminidase did not alter the parasite negativeness. Cytochemically, we could observe a reduction in the cationized ferritin binding after the parasite treatment with each of the former enzymes, but not with neuraminidase. The surface free energy of parasites was calculated by taken to account the London dispersion, the Keeson dipole-dipole, and the Debye dipole-induced forces, as well as the surface polarity of the parasites and their zeta-potentials, by considering their adhesion to polystyrene surfaces. The delta G values of -6.4 and -18.1 mJ.m-2 were obtained for avirulent and virulent promstigotes, respectively.     

Geographic differences in the allele frequencies of the human Y-linked tetranucleotide polymorphism DYS19

We have studied the allele frequency distribution of the microsatellite locus DYS 19 in several populations with different geographical origins worldwide. Three new alleles were found. In addition, remarkable geographic and ethnic differences were observed in the allele frequency profiles and DNA marker (gene) diversity among populations and major ethnic groups. Amerindians showed an overwhelming predominance of the A allele, while in Caucasians the B allele was modal, and in Greater Asians and Africans allele C became predominant. Even within these geographic regions there were significant gradients, as exemplified by the decreasing frequency profile of the B allele from Great Britain over Germany to Slovakia. Thus, DYS 19 emerges as a useful tool for studying the structure and dynamics of human populations.     

The magic-bead concept: an integrated approach to nitrogen removal with co-immobilized micro-organisms

This paper describes both qualitative and quantitative aspects of simultaneous autotrophic nitrification and heterotrophic denitrification by, respectively, the nitrifierNitrisomonas europaea and either of the denitrifiersPseudomonas denitrificans orParacoccus denitrificans co-immobilized in double-layer gel beads. The system is based on the establishment of well-defined oxic and anoxic zones within the cell supports and on physical separation of the nitrifying and denitrifying populations. Nitrification and denitrification rates were obtained from measured bulk concentrations and head-space analysis. The latter analyses showed that ammonia was primarily converted into molecular nitrogen. Nitrous oxide was not detected. High nitrogen removal rates (up to 5.1 mmol N m^–3 gel s^–1) were achieved in continuous reactors under aerobic conditions. The overall rate of nitrogen removal was controlled by the nitrifying step. The approach followed is, in principle, also suitable to the coupling of other oxidative and reductive bioprocesses having complementary metabolic routes. Two-stage bioconversion processes can be thus conducted as if single-staged, which results in more compact reactor systems.