Primary role of NADH formed by glucose dehydrogenase in ATP provision at the early stage of spore germination in Bacillus megaterium QM B1551

Metabolic events involved in energy metabolism were studied in order to evaluate the ATP-forming ability of Bacillus megaterium QM B1551 spores at the very early stage of germination. When heat-activated spores were germinated on glucose as a sole substrate, its oxidation into gluconate (catalyzed by glucose dehydrogenase, EC 1.1.1.47), the accompanying NADH formation, oxygen uptake, and RNA synthesis were initiated immediately after germination, even when anaerobic breakdown of 3-phosphoglycerate (an ATP source for spores) and the subsequent glucose metabolism via the phosphorylating pathway were impaired by potassium fluoride (KF). In contrast, fructose metabolism and the accompanying metabolic events did not begin until a few minutes after triggering of germination, and those events were entirely abolished by KF, indicating that fructose metabolism is initiated exclusively via its phosphorylation by the ATP derived from endogenous 3-phosphoglycerate. Thus those results provided further evidence for our previous proposal (Otani et al (1987) Microbiol. Immunol. 31: 967-974; Sano et al (1988) Biochem. Biophys. Res. Commun. 151: 48-52) that the first molecules of ATP in germinating spores can be efficiently generated via aerobic oxidation of NADH, which is formed by glucose dehydrogenase. Fluorescence monitoring of NADH in germinating spores also supported this conclusion.     

Sensitivity in Detection of Monoclonal Immunoglobulins by Thin-Layer Isoelectric Focusing and Certain Electrophoreses in Laboratory Use

By applying sera containing various myeloma proteins to thin-layer isoelectric focusing at various pH ranges, we found that the best range of pH gradient for the detection of the band group specific for monoclonal immunoglobulin was from 5 to 8. The lowest concentration of monoclonal immunoglobulin detected by this method was approximately 0.1 mg/ml which was 8 to 32 times lower than the concentration detectable by cellulose acetate membrane electrophoresis or immunoelectrophoresis in routine laboratory use. Monoclonal protein concentration from 0.1 to 20 mg/ml was determined quantitatively. Blind tests on sera resulted in a disagreement between the concentration of monoclonal immunoglobulin assumed from the results of cellulose acetate membrane electrophoresis and immunoelectrophoresis and that determined by isoelectric focusing. These results suggest that isoelectric focusing is useful for the surveillance of monoclonal immunoglobulinemia provided that the technique and equipment are improved for laboratory tests.     

Examining the genetic diversity of the orange blossom wheat midge,Sitodiplosis mosellana (Géhin), across North America

The orange blossom wheat midge, Sitodiplosis mosellana (Géhin) (Diptera: Cecidomyiidae), is a significant pest of wheat (Triticum spp.) grown in the Northern Hemisphere. It was accidently introduced to North America over 200 years ago and has subsequently spread throughout the northern Great Plains. Since 2010, several Canadian spring wheat varieties containing the resistance gene Sm1 have been released. Due to the potential of wheat midge populations to evolve virulent biotypes to Sm1, cultivars containing Sm1 are grown with a susceptible cultivar in an interspersed refuge. An understanding of the genetic diversity of wheat midge populations could provide important information on the potential development of resistance to Sm1. In the current study, we used two mitochondrial genes (CO1 and ND4) from wheat midge collected across the northern Great Plains and Québec in North America to assess population structure and genetic diversity. We found limited genetic diversity and population structure across the sampled North American populations. We also assessed North American haplotype similarity to wheat midge collected from Europe and China and found high similarity between North American and European populations, although sampling in Europe was limited. This supports the hypothesis that North American populations originated from Europe.     

Glucose catabolism during germination of Bacillus megaterium spores.

When heat-activated spores of Bacillus megaterium germinated in glucose-containing medium, 10 to 30% of the glucose was found to be oxidized to gluconate.     

Conformational preference of bicyclic β-amino acid dipeptides

Bridged bicyclic amino acids have high potential applicability as self-organized, conformationally constrained synthetic building blocks that do not require assistance from hydrogen bond formation. We systematically investigated the intrinsic conformational propensities of dipeptides of bridged bicyclic β-amino acids by means of accelerated molecular dynamics simulation and density functional theory (DFT) calculations in methanol, chloroform, and water. While the main-chain conformation, represented by φ and θ values, is fixed by the nature of the bicyclic ring structure, rotation of the C-terminal carbonyl group (ψ) is also restricted, converging to one or two minima. In endo-type dipeptides, in which the two N- and C-terminal amides are spatially close to each other, the C-terminal amide plane is placed horizontally. In exo-type dipeptides, in which the two amides are on opposite sides of the ring plane, the C-terminal carbonyl group can take two types of positions: either parallel/antiparallel with the N-terminal carbonyl or beneath the bicyclic ring, forcing the amide NHMe moiety to lie outside of the ring. We also examined the cis-trans preference of model bicyclic amides. Although the parent amides exhibit cis-trans equilibrium without any preference, addition of a methyl group on one of the bridgehead positions tips the equilibrium towards trans.     

Phorbol esters stimulate spermidine/spermine N1-acetyltransferase activity in mitogen-stimulated bovine lymphocytes

Phorbol 12-myristate-13-acetate (PMA) is shown to induce spermidine/spermine N1-acetyltransferase, a rate-limiting enzyme of polyamine biodegradation, in bovine lymphocytes. When PMA and phytohemagglutinin (PHA) were added simultaneously, the enzyme activity was stimulated synergistically. The ability of phorbol esters to stimulate the enzyme activity was consistent with their tumor-promoting ability. Phorbol, which is not a tumor promotor, was incapable of stimulating the enzyme activity. Phorbol diacetate weakly stimulated the activity of the acetylase. Phorbol dibutyrate had a similar stimulatory effect to PMA. These results suggest that the spermidine/spermine N1-acetyltransferase may play an important role in changes in polyamine levels in phorbol ester-treated cells and that the increase in the enzyme activity may have some relationship to the control of cell growth and differentiation by phorbol esters.