Nutrient and chemical sensing by intestinal pathogens

Pathogenic gut bacteria, such as those comprising the Enterobacteriaceae family, have evolved sophisticated virulence mechanisms, including nutrient and chemical sensing, to escape host defense strategies and produce disease. In this review we describe the mechanisms utilized by the enteric pathogen enterohemorrhagic Escherichia coli (EHEC) O157:H7 to achieve successful colonization of its mammalian host.     

Fractal root system of Melica przewalskyi along different aspect in degraded grassland

Aims Fractal root system is phenotypic plasticity result of plant root architecture to respond to environmental heterogeneity, may reflect the growth strategy of plants to adapt to environmental conditions. Our objective was to explore the relationship between root fractal dimension and fractal abundance of fractal root system of Melica przewalskyi population in response to aspect variation in the northwest of China. Methods The study site was located in a degraded alpine grassland on the northern slope in Qilian Mountains, Gansu Province, China. Survey and sampling were carried out at 40 plots which were set up along four slope aspects transects with 20 m distance between adjacent plots. Handheld GPS was used to determine the elevation, longitude and latitude of each plot. ArcGIS was used to set up digital elevation model (DEM). Community traits were investigated and six individuals roots of M. przewalskyi were collected randomly at each plot. The samples were cleaned and divided into different organs, then scanning the root with the Win-RHIZO for measurements of fractal dimension and fractal abundance in laboratory, and their biomass were then measured after being dried at 80 °C in an oven. Important findings With the slope aspect turned from north to east, west, and south, the density, height and soil moisture content of the plant community displayed a pattern of initial decline, the height, density, root fractal abundance of M. przewalskyi increased and the root fractal dimension decreased. The root fractal dimension was negatively associated with the fractal abundance in all aspects, but the relationship varied along the slope aspects gradient; there was a highly significant negative correlation (p < 0.01) between the root fractal dimension and fractal abundance at north slope and south slope aspect, whereas the correlation only reached a significant level (p < 0.05) at the east slope aspect and west slope aspect; indicating that there is a trade-off between the root fractal dimension and fractal abundance. In addition, when the slope aspect changed from north to east, west and south, the standardized major axis (SMA) slope of the regression equation in the scaling relationships between root fractal dimension and fractal abundance increased (p < 0.05), indicating that the roots of M. przewalskyi at the droughty southern slope have less branch and more sparse in the same soil volume of root exploitation and utilization. Consequently, the resource allocation pattern on reasonable trade-off between root fractal dimension and fractal abundance in different slope aspect of M. przewalskyi, reflects the relationship between the income and the cost of construction of plant root architecture.     

Inhibitory effect of aspirin on cholera toxin-induced phospholipase and cyclo-oxygenase activity

Cholera toxin (CT) stimulated phospholipase activity and caused [3H]arachidonic acid (3H-AA) release in a murine macrophage/monocyte cell line. Pretreatment of cells with dexamethasone, a phospholipase A2 (PLA2) inhibitor, did not affect CT-induced 3H-AA release. In contrast, aspirin, which is an inhibitor of phospholipase C (PLC), blocked CT-induced 3H-AA release and subsequent prostaglandin (PC) synthesis. The inhibitory effect of aspirin was dose dependent, with 4 mM reducing the CT response by approximately 50%. Similarly, inhibition was time dependent, occurring when the drug was added to the culture medium as late as 30 min after CT. Brief exposure (30 min) of the cells to aspirin did not alter their subsequent response to CT, but 3H-AA release from cells exposed to aspirin for 2.5 h was irreversibly inhibited. The data suggested that CT stimulation of AA metabolism may involve increased PLC activity.     

Localization of Endo-Oligopeptidase (EC 3.4.22.19) in the Rat Nervous Tissue

The subcellular and regional distribution of endo-oligopeptidase (EC 3.4.22.19), an enzyme capable of generating enkephalin by single cleavage from enkephalin-containing peptides, was determined by an enzymatic assay using metorphamide and by immunochemical techniques in the CNS of the rat. The rat CNS contains a membrane-associated form of endo-oligopeptidase, an enzyme predominantly associated with the soluble fraction of brain homogenates. Subcellular fractionation showed that approximately 17% of the total activity of the enzyme is associated with membrane fractions including synaptosomes. Synaptosomal membranes were prepared from neocortex, striatum, hypothalamus, medulla, spinal cord, and cerebellum. The amount of EC 3.4.22.19 activity solubilized by 3-[( 3-cholamidopropyl]dimethylammonio)-1-propanesulfonate from synaptosomal membranes was similar in neocortex, striatum, and hypothalamus, being three- to 10-fold greater than in spinal cord, cerebellum, and medulla. A polyclonal antibody exhibiting high affinity for endo-oligopeptidase was raised in rabbits against the purified rat brain enzyme and used to localize endo-oligopeptidase by Western blotting and by immunoperoxidase techniques. A strong band corresponding to the Mr of EC 3.4.22.19 was found in solubilized proteins obtained from synaptosomal membranes prepared from hypothalamus, neocortex, and striatum when subjected to Western blotting. The immunohistochemical localization of endo-oligopeptidase indicated that the immunoreactivity was confined to gray matter in regions known to be rich in peptide-containing neurons such as the striatum. In the cerebellum, a region poor in peptides, no staining could be detected. The nonuniform distribution of endo-oligopeptidase in rat brain suggests a role in neurotransmitter processing in the CNS.     

Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Observations on lesser-known flatworms: temnocephala

Temnocephala novae-zealandie, a flatworm epizoic on crayfish, was examined by light and electron microscopy to investigate the variation within rhabdocoel turbellarians and to provide information on the possible structural modifications in the evolution of parasitism. The sucker is clearly glandular; the tentacular glands are eosinophilic and at the surface store and often release mucus as a coiled thread; the epidermis is clearly not reduced in structure and contains septate junctions in the anterior portion. Light microscope studies documented the presence of ten pair of paranephrocytes (athrocytes). In the laboratory, egg deposition, survival on and apart from the host and another crayfish, and behavior during the host molt were observed.