Thigmomorphogenesis: The induction of callose formation and ethylene evolution by mechanical perturbation in bean stems

Mechanical perturbation by rubbing of the first internode of 11–12 day old plants of Phaseolus vulgaris L. cv. Cherokee wax induces the rapid deposition of callose in the cells of phloem and other tissues. Callose deposition begins immediately after mechanical perturbation, and shows a minor transient peak 1.5 h, and a major peak 6 h later. The callose gradually disappears and is gone after 3 days. If the stems are perturbed every day, the amount of callose decreases by day 2 but then gradually increases again through day 12. Both the top and bottom of the internode produce callose in response to mechanical perturbation. The evolution of ethylene in response to mechanical perturbation begins after 1 h, peaks at 2–3 h and is gone by 5–6 h. A spray of 10⁻² M 2-deoxy-D-glucose (DDG) completely blocks stem thickening, callose deposition and ethylene evolution due to mechanical perturbation. DDG at 10⁻⁵ to 10⁻⁴ M blocks callose production in mechanically perturbed stem segments and increases ethylene evolution from unperturbed stem segments to greater levels than those obtained by mechanically perturbed segments. It is concluded that mechanical perturbation of bean stems tissue induces deposition of callose more rapidly than it induces evolution of ethylene and that DDG can block both processes.     

Linkage Relationships Reflecting Ancestral Tetraploidy in Salmonid Fish

Fifteen classical linkage groups were identified in two salmonid species (Salmo trutta and Salmo gairdneri) and three fertile, interspecific hybrids (S. gairdneri X Salmo clarki, Salvelinus fontinalis X Salvelinus namaycush and S. fontinalis X Salvelinus alpinus) by backcrossing multiply heterozygous individuals. These linkage relationships of electrophoretically detected, protein coding loci were highly conserved among species. The loci encoding the enzymes appeared to be randomly distributed among the salmonid chromosomes. Recombination frequencies were generally greater in females than in males. In males, certain linkage groups were pseudolinked with other linkage groups, presumably because of facultative multivalent pairing and directed disjunction of chromosomes. Five such pseudolinkage groups were identified and they also appeared to be common among species and hybrids. Duplicate loci were never classically linked with each other, although some exhibited pseudolinkage and some showed evidence of exchanging alleles. Gene-centromere recombination frequencies estimated from genotypic distributions of gynogenetic offspring were consistent with map locations inferred from female intergenic recombination frequencies. These linkage relationships support the contention that all extant salmonids arose from a common tetraploid progenitor and that this progenitor may have been a segmental allotetraploid.     

Antimicrobial activity of lysozyme against bacteria involved in food spoilage and food-borne disease.

Egg white lysozyme was demonstrated to have antibacterial activity against organisms of concern in food safety, including Listeria monocytogenes and certain strains of Clostridium botulinum. We also found that the food spoilage thermophile Clostridium thermosaccharolyticum was highly susceptible to lysozyme and confirmed that the spoilage organisms Bacillus stearothermophilus and Clostridium tyrobutyricum were also extremely sensitive. Several gram-positive and gram-negative pathogens isolated from food poisoning outbreaks, including Bacillus cereus, Clostridium perfringens, Staphylococcus aureus, Campylobacter jejuni, Escherichia coli O157:H7, Salmonella typhimurium, and Yersinia enterocolitica, were all resistant. The results of this study suggest that lysozyme may have selected applications in food preservation, especially when thermophilic sporeformers are problems, and as a safeguard against food poisoning caused by C. botulinum and L. monocytogenes.     

Evaporative Cooler Use Influences Temporal Indoor Relative Humidity but Not Dust Mite Allergen Levels in Homes in a Semi-Arid Climate

Concerns about energy consumption and climate change make residential evaporative coolers a popular alternative to central air conditioning in arid and semi-arid climates. However, evaporative coolers have been shown to significantly increase indoor relative humidity and dust mite allergen levels in some studies, while showing no association in other studies. Improved measurement of temporal fluctuations in indoor relative humidity may help identify factors that promote mite growth in homes in dry climates. Dust samples and continuous indoor relative humidity measurements were collected from homes with central air conditioning and homes with evaporative coolers in Utah. Samples were collected over two seasons, winter/spring (Jan–Apr) and summer (July–Sept), 2014. Dust samples were analyzed for Der p 1 and Der f 1 using a two-site monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) analysis. Housing characteristics including age of home, occupant density, and age of mattresses, furniture, and carpeting were also measured. Positive Der p 1 or Der f 1 samples were found in 25.0% of the homes and there was no difference in mean allergen levels by type of air conditioning. Indoor relative humidity was significantly higher in homes with evaporative coolers compared to those with central air conditioning during the summer. Homes with evaporative coolers also spent significantly more time during summer above 55.0% and 65.0% relative humidity compared to central air homes, but not above 75.0%. Findings from this study suggest that increased humidity from evaporative coolers may not be sufficient to exceed the critical equilibrium humidity or maintain humidity excursions for sufficient duration in relatively larger single-family homes in semi-arid climates to support mite growth and reproduction.     

Critical micelle concentrations of gangliosides

The micellar properties of mixed, bovine gangliosides and purified galactosyl-N-acetylacetylgalactosaminyl (N-acetylneuraminyl) galactosylglucosylceramide were studied by gel filtration, equilibrium dialysis, and band and boundary centrifugation in sucrose gradients. The dissociation of micelles is very slow (days) in water and required us to approach equilibrium by association of monomers rather than by the dissociation of micelles. The gangliosides were therefore first converted into very low molecular weight aggregates (1-3 molecules) by dissolving them in Me2SO. Galactosyl-N-acetylgalactosaminyl(N-acetylneuraminyl)galactosylglucosylceramide was then diluted into aqueous sucrose gradients and sedimented by the boundary centrifugation technique. This gave a sedimenting micelle and a nonsedimenting monomer concentration of (1-2) x 10-10 M (or less) which corresponds to the critical micelle concentration value. The mixed gangliosides revealed two micellar sizes (i.e., 10 and 4.5 S), the slower sedimenting species being formed from the larger one with time (days). The critical micelle concentration of the mixed gangliosides was found to be approximately 10-8 M by a gel filtration, equilibrium dialysis, and band centrifugation.     

The role of IFN in respiratory syncytial virus pathogenesis

Formalin-inactivated respiratory syncytial virus (RSV) vaccine preparations have been shown to cause enhanced disease in naive hosts following natural infection. In this study we demonstrate a similar pattern of enhanced disease severity following primary RSV infection of IFN-nonresponsive STAT1(-/-) mice. STAT1(-/-) mice showed markedly increased illness compared with wild-type BALB/c animals following RSV inoculation despite similar lung virus titers and rates of virus clearance. Histologically, STAT1(-/-) animals had eosinophilic and neutrophilic pulmonary infiltrates not present in wild-type or IFN-gamma(-/-)-infected mice. In cytokine analyses of infected lung tissue, IFN-gamma was induced in both STAT1(-/-) and wild-type mice, with preferential IL-4, IL-5, and IL-13 induction only in the STAT1(-/-) animals. Eotaxin was detected in the lungs of both wild-type and STAT1(-/-) mice following infection, with a 1.7-fold increase over wild-type in the STAT1(-/-) mice. Using a peptide epitope newly identified in the RSV fusion protein, we were able to demonstrate that wild-type memory CD4(+) T cells stimulated by this peptide produce primarily IFN-gamma, while STAT1(-/-)CD4(+) cells produce primarily IL-13. These findings suggest that STAT1 activation by both type I (alphabeta) and type II (gamma) IFNs plays an important role in establishing a protective, Th1 Ag-specific immune response to RSV infection.     

Design and in vivo immunogenicity of a polyvalent vaccine based on SIVmac regulatory genes

Most vaccine modalities for human immunodeficiency virus type 1 (HIV-1) tested for immunogenicity and efficacy in the SIVmac (simian immunodeficiency virus) macaque model do not include the viral regulatory proteins. Because viral regulatory proteins are expressed early during the virus life cycle and represent an additional source of antigens, their inclusion as a vaccine component may increase the overall virus-specific immune response in vaccinees. However, at least two of the early proteins, Tat and Nef, may be immunosuppressive, limiting their usefulness as components of an SIV vaccine. We have constructed a polyvalent chimeric protein in which the open reading frames for Tat and Nef have been reassorted and the nuclear localization sequence for Tat and Rev and the myristoylation site for Nef have been removed. The resulting DNA plasmid (pDNA-SIV-Retanef) (pDNA-SIV-RTN) encodes a protein of 55 kDa (Retanef) that localizes at the steady state in the cytoplasma of transfected cells. Both the DNA-SIV-RTN and the highly attenuated recombinant poxvirus vector NYVAC-SIV-RTN were demonstrated to be immunogenic in SIVmac251-infected macaques treated with ART as well as in naive macaques. An equivalent strategy may be used for the generation of polyvalent antigens encoding the regulatory proteins in a HIV-1 vaccine candidate.     

Phytate as the sole phosphate source for growth of Tetrahymena

Phytate, the storage form of phosphate in seeds and grains, is a major form of environmental phosphate loading from fertilizer inputs and agricultural runoff. We have investigated the ability of Tetrahymena populations to grow on phytate as their sole phosphate source. Populations grew equally well in chemically defined medium with phosphate and medium in which the phosphate was replaced with phytate in comparable concentrations between 0.5 mM and 6 mM. Intracellular phytate concentrations of cells grown in phytate showed a 4-6-fold increase over those grown in phosphate when measured during the late stage of exponential growth. These results demonstrate that phytate can provide a source of adequate phosphate for sustained growth in phytate-rich environments.