Conjugative plasmids in Neisseria gonorrhoeae.

A conjugation system initially discovered in beta-lactamase-producing gonococci mobilized small non-selftransmissible R plasmids encoding beta-lactamase (penicillinase) production into other gonococci, Neisseria, and Escherichia coli. This conjugation system was mediated by a separate selftransmissible plasmid of 23.9 X 10(6) daltons, pFA2. Conjugative plasmids capable of mobilizing R plasmids were also found in nearly 8% of the non-penicillinase-producing gonococci. These were similar to pFA2 in size, buoyant density, and restriction endonuclease digest patterns but were less efficient than pFA2 in mobilization of the penicillinase plasmid pFA3. The presence of conjugative plasmids in gonococci isolated before the appearance of penicillinase-producing strains indicates that a conjugation system for plasmid transfer predated the appearance of R plasmids in gonococci.     

Studies of growth and development of gametocytes in Haemoproteus columbae Kruse.

As determined by their ability to exflagellate and round up, it took the macrogametocyte and the microgametocyte of Haemoproteus columbae in pigeons 68 and 116 h, respectively, after patency to reach maturity. Pigment granules appeared in the undifferentiated gametocytes 8 h after invasion of blood. Vacuoles were observed in young gametocytes and persisted in the older forms. The growth curve of H. columbae is close to the sigmoidal curve for growth in protozoa. Multiple infection was noticed in pigeons with high levels of parasitemia, but no more than 2 gametocytes reached maturity; such multiple infections were rare in relapses. The sex ratio of the gametocytes was 1:1. Strong lateral displacement of the nuclei of infected erythrocytes was the rule; hypertrophy was negligible.     

Edaphic factors and plant species distribution in a protected area in the desert of Bahrain Island

Effects of edaphic factors (salinity, pH, Na⁺, K⁺, Ca⁺⁺, CaCO₃, water holding capacity, and grain size) on the spatial distribution of plants were investigated. Soil was sampled at 22 stands. Sixteen plant species were recorded from these stands. Relation between edaphic factors and plant distribution was investigated using correlation statistical analysis. Distribution of some plants was found to be highly correlated with edaphic factor(s).     

Effect of N,N-Dicyclohexylcarbodiimide on Acetylcholine Release from Torpedo Synaptosomes and Proteoliposomes Reconstituted with the Proteolipid Mediatophore

The mediatophore is a presynaptic membrane protein that has been shown to translocate acetylcholine (ACh) under calcium stimulation when reconstituted into artificial membranes. The mediatophore subunit, a 15-kDa proteolipid, presents a very high sequence homology with the N,N'-dicyclohexylcarbodiimide (DCCD)-binding proteolipid subunit of the vacuolar-type H(+)-ATPase. This prompted us to study the effect of DCCD, a potent blocker of proton translocation, on calcium-dependent ACh release. The present work shows that DCCD has no effect on ACh translocation either from Torpedo synaptosomes or from proteoliposomes reconstituted with purified mediatophore. However, using [14C]DCCD, we were able to demonstrate that the drug does bind to the 15-kDa proteolipid subunit of the mediatophore. These results suggest that although the 15-kDa proteolipid subunits of the mediatophore and the vacuolar H(+)-ATPase may be identical, different domains of these proteins are involved in proton translocation and calcium-dependent ACh release and that the two proteins have a different membrane organization.     

Mobilization of non-exchangeable K by ryegrass in five Moroccan soils with and without mica

Intensive cropping of Italian ryegrass (Lolium multiforum L.) in pots was used to assess the contribution of non-exchangeable K to plant uptake. The soils used were: two soils high in mica (illite) developed on recent alluvium plus two smectitic (beidellitic) soils and a soil of mixed mineralogy rich in mica. Four K treatments were used (0, 28.6, 143, and 286 mg kg^-1 soil) with 8 successive monthly cuttings. A response of plant K uptake to added K was observed in all soils. Both 1.0 M NH₄0Ac and 0.2 M CaCl₂ extractable K were depleted to a minimum level specific for each soil. The minima were lower in the old upland soils compared to the young alluvial soils. Uptake of K by Italian ryegrass induced K release from the non-exchangeable K to replenish the plant available pool of K ions. The release of mica interlayer K in the alluvial and in the high K smectitic soil supplied sufficient K to plants even under intensive cropping. The rate of mobilization of interlayer K was low in the smectitic soil with lower K. The lowest release rate was in the old high mica soil. Iron coatings may have inhibited mobilization of interlayer K. The rates of mobilization cannot be predicted from mineralogical and K-extraction data only. The rates of K uptake and the rates of K release by ryegrass under intensive cropping are potential values which can be used for modelling K availability to plants in the soils studied.     

Cation transport mechanisms in Mycoplasma mycoides var. Capri cells. The nature of the link between K+ and Na+ transport

We have studied the links between the mechanisms of Na(+), K(+) and H(+) movements in glycolysing Mycoplasma mycoides var. Capri cells. In the light of the results reported in the preceding paper [Benyoucef, Rigaud & Leblanc (1982) Biochem. J.208, 529-538], we investigated certain properties of the membrane-bound ATPase of Mycoplasma cells, with special reference to its ionic requirements and sensitivity to specific inhibitors. Our findings show, first, that, although Na(+) stimulated ATPase activity, K(+) did not affect it, and, secondly, that NN'-dicyclocarboidi-imide and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD) were potent inhibitors of the basal ATPase activity, which was unaffected by vanadate and ouabain. We also investigated the movements of Na(+) and H(+) under the experimental conditions applied to the study of the K(+) uptake reported in the preceding paper, and found that when ;Na(+)-loaded cells' previously equilibrated with (22)Na(+) were diluted in a sodium-free medium, addition of glucose induced a rapid efflux of (22)Na(+). This energy-dependent efflux was independent of the presence of KCl in the medium. Studies of the changes in internal pH by 9-aminoacridine fluorescence or [(14)C]methylamine distribution indicated that the movement of Na(+) was coupled to that of protons moving in the opposite direction, a finding that supports the presence of an Na(+)/H(+) antiport. When Na(+)-loaded cells are diluted in an Na(+)-rich medium the Na(+)/H(+) antiport is still active, but cannot decrease the intracellular Na(+) concentration. Under such conditions, net (22)Na(+) extrusion is specifically dependent on the presence of K(+) in the medium. The present results and those derived from the study of K(+) accumulation (the preceding paper) can be rationalized by assuming that Mycoplasma mycoides var. Capri cells contain two transport systems for Na(+) extrusion: an Na(+)/H(+) antiport and an ATP-consuming Na(+)/K(+)-exchange system.     

Preparation and characterization of detoxified lipopolysaccharide-protein conjugates

Alkaline treatment of Pseudomonas aeruginosa type 5 lipopolysaccharide (LPS) resulted in reduced toxicity as measured by both the Limulus amoebocyte assay and the rabbit pyrogenicity test. Chemical analysis of the deacylated LPS (D-LPS) revealed that ester-linked fatty acids were removed while the amide-linked fatty acids remained intact. The neutral and amino sugar compositions for native LPS and D-LPS were identical within experimental error. Antigenic determinants for complement-dependent human opsonic antibody were retained under these deacylation conditions. To enhance its immunogenicity, D-LPS was covalently coupled to Pseudomonas pili and the 1,4-diaminobutyl derivatives of Pseudomonas exotoxin A and tetanus toxoid. Quantitative amino sugar analyses revealed that 2.6 and 3.2 mol of D-LPS were covalently bound to aminobutyl Pseudomonas exotoxin A and aminobutyl tetanus toxoid, respectively. Gel electrophoresis data indicated at least 1 mol of D-LPS covalently bound per pilus subunit protein. Initial immunologic data indicated that antibody against D-LPS could be induced when the D-LPS is covalently attached to protein carriers.     

Studies on aroyl- and aryl-hydrazide derivatives from d-glycero-d-gulo-heptono-1,4-lactone

A series of aroyl- and aryl-hydrazide derivatives was prepared from d-glycero-d-gulo-heptono-1,4-lactone (1). The reactivity of the NH proton in these hydrazides, in terms of their dissociation constants (pK_a), was determined from their electronic spectra, and correlated to the Hammett σ values of the substituents. Comparable reactivities of the NH protons for the compounds, and the effect of the substituent, were studied by n.m.r. spectroscopy. Decomposition of the aroylhydrazides with copper(II) sulfate or nitrous acid resulted in the regeneration of 1.     

Complexes of vitamin B6, part VII: Ternary complexes of cobalt(II), nickel(II), and copper(II) involving pyridoxamine and some amino acids

The stability constants of the ternary Cu(II), Ni(II), and Co(II) complexes containing pyridoxamine (PM) and as a second ligand (L) glycine, DL-alanine, DL-valine, and β-phenylalnine were determined by pH-metric titration in 0.50 M KNO₃ at 30°C. The corresponding constants of the equilibrium, log X, are greater than would be expected for purely statistical reasons (log X = 0.6), except for few complex cases of Co(II). It has been also concluded that amino acids compete more than pyridoxamine for Ni(II) and Co(II) through the formation of 1:2:1:0 species rather than 2:1:1:0 of PM⁻:L⁻:M²⁺:H⁺.     

Synthesis of 2-acetamido-2-deoxy-3-O-β-d-mannopyranosyl-d-glucose

Condensation of 4,6-di-O-acetyl-2,3-O-carbonyl-α-d-mannopyranosyl bromide with benzyl 2-acetamido-4,6-O-benzylidene-2-deoxy-α-d-glucopyranoside (2) gave an α-d-linked disaccharide, further transformed by removal of the carbonyl and benzylidene groups and acetylation into the previously reported benzyl 2-acetamido-4,6-O-benzylidene-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl-α-d-mannopyranosyl)-α-d-glucopyranoside. Condensation of 3,4,6-tri-O-benzyl-1,2-O-(1-ethoxyethylidene)-α-d-glucopyranose or 2-O-acetyl-3,4,6-tri-O-benzyl-α-d-glucopyranosyl bromide with 2 gave benzyl 2-acetamido-3-O-(2-O-acetyl-3,4,6-tri-O-benzyl-β-d-glucopyranosyl)-4,6-O-benzylidene-2-deoxy-α-d-glucopyranoside. Removal of the acetyl group at O-2, followed by oxidation with acetic anhydride-dimethyl sulfoxide, gave the β-d-arabino-hexosid-2-ulose 14. Reduction with sodium borohydride, and removal of the protective groups, gave 2-acetamido-2-deoxy-3-O-β-d-mannopyranosyl-d-glucose, which was characterized as the heptaacetate. The anomeric configuration of the glycosidic linkage was ascertained by comparison with the α-d-linked analog.