Cultivation of Mesophilic Soil Crenarchaeotes in Enrichment Cultures from Plant Roots

Because archaea are generally associated with extreme environments, detection of nonthermophilic members belonging to the archaeal division Crenarchaeota over the last decade was unexpected; they are surprisingly ubiquitous and abundant in nonextreme marine and terrestrial habitats. Metabolic characterization of these nonthermophilic crenarchaeotes has been impeded by their intractability toward isolation and growth in culture. From studies employing a combination of cultivation and molecular phylogenetic techniques (PCR-single-strand conformation polymorphism, sequence analysis of 16S rRNA genes, fluorescence in situ hybridization, and real-time PCR), we present evidence here that one of the two dominant phylotypes of Crenarchaeota that colonizes the roots of tomato plants grown in soil from a Wisconsin field is selectively enriched in mixed cultures amended with root extract. Clones recovered from enrichment cultures were found to group phylogenetically with sequences from clade C1b.A1. This work corroborates and extends our recent findings, indicating that the diversity of the crenarchaeal soil assemblage is influenced by the rhizosphere and that mesophilic soil crenarchaeotes are found associated with plant roots, and provides the first evidence for growth of nonthermophilic crenarchaeotes in culture.     

Spatial and temporal structures of pheromone plumes in fields and forests

Wind‐borne odour stimuli from a small point‐source of pheromone are intermittent owing to the effects of atmospheric turbulence on the odour plume. The work reported here measures the characteristics of the intermittent stimulus in open fields and forests during typical daytime atmospheric conditions. To model the pheromone plume, negatively charged atmospheric ions were used as a tracer. They were released from an ion generator, and ion detectors measured the fluctuating flux of ions at positions up to 20 m downwind in the open field case and 10 m in the forest. In both the open field and in the forest, ion signals were highly intermittent, with a signal present only 20% of the time. Ion signals recorded in the forest consisted of bursts with gaps between them of at least three‐fold greater duration than those from the open field. In both environments, bursts generally each comprised a series of ‘spikes’, on average three in the field and seven in the forest. To validate the use of ionized air plumes as models of pheromone plumes, the antennae of male Lymantria dispar (gypsy moth) were used as detectors to quantify the plume of synthetic (+)‐disparlure emanating from a 2000 ng point source placed ≈ 10 cm from the ion source. A comparison of ion signals and EAGs (electronantennograms) suggests that the antennae respond to the main spikes within a burst, but no consistent relationship between the strength of the spikes and the magnitude of the EAG response was found. The average strengths of bursts in the ion detector signal decreased systematically as the distance from the ion generator to the ion detector increased. A similar trend, however, was not detected in the EAG response.     

The involvement of microRNAs in Type 2 diabetes

T2D (Type 2 diabetes mellitus) is a major health issue that has reached epidemic status worldwide. T2D is a progressive metabolic disorder characterized by reduced insulin sensitivity, insulin resistance and pancreatic β-cell dysfunction. Improper treatment of TD2 can lead to severe complications such as heart disease, stroke, kidney failure, blindness and nerve damage. The aetiology and molecular mechanisms of T2D are not fully understood, but compelling evidence points to a link between T2D, obesity, dyslipidaemia and insulin resistance. Although T2D seems to be strongly linked to environmental factors such as nutrition and lifestyle, studies have shown that genetic factors, such as polymorphisms associated with metabolic genes, imprinting, fetal programming and miRNA (microRNA) expression, could also contribute to the development of this disease. miRNAs are small 22-25-nt-long untranslated RNAs that negatively regulate the translation of mRNAs. miRNAs are involved in a large number of biological functions such as development, metabolism, immunity and diseases such as cancer, cardiovascular diseases and diabetes. The present review examines the various miRNAs that have been identified as being potentially involved in T2D, focusing on the insulin-sensitive organs: white adipose tissue, liver, skeletal muscle and the insulin-producing pancreatic β-cells.     

Obestatin acts in brain to inhibit thirst

Derived from the same prohormone, obestatin has been reported to exert effects on food intake that oppose those of ghrelin. The obestatin receptor GPR39 is present in brain and pituitary gland. Since the gene encoding those two peptides is expressed also in those tissues, we examined further the possible actions of obestatin in vivo and in vitro. Intracerebroventricular administration of obestatin inhibited water drinking in ad libitum-fed and -watered rats, and in food-and water-deprived animals. The effects on water drinking preceded and were more pronounced than any effect on food intake, and did not appear to be the result of altered locomotor/behavioral activity. In addition, obestatin inhibited ANG II-induced water drinking in animals provided free access to water and food. Current-clamp recordings from cultured, subfornical organ neurons revealed significant effects of the peptide on membrane potential, suggesting this as a potential site of action. In pituitary cell cultures, log molar concentrations of obestatin ranging from 1.0 pM to 100 nM failed to alter basal growth hormone (GH) secretion. In addition, 100 nM obestatin failed to interfere with the stimulation of GH secretion by GH-releasing hormone or ghrelin and did not alter the inhibition by somatostatin in vitro. We conclude that obestatin does not act in pituitary gland to regulate GH secretion but may act in brain to alter thirst mechanisms. Importantly, in rats the effects of obestatin on food intake may be secondary to an action of the peptide to inhibit water drinking.     

A robust and cost-effective method for the production of Val, Leu, Ile (δ1) methyl-protonated 15N-, 13C-, 2H-labeled proteins

A selective protonation strategy is described that uses [3-2H] 13C -ketoisovalerate to introduce (1H- methyl)-leucine and (1H- methyl)-valine into 15N-, 13C-, 2H-labeled proteins. A minimum level of 90% incorporation of label into both leucine and valine methyl groups is obtained by inclusion of 100 mg/L -ketoisovalerate in the bacterial growth medium. Addition of [3,3-2H2] -ketobutyrate to the expression media (D2O solvent) results in the production of proteins with (1H-1 methyl)-isoleucine (>90% incorporation). 1H-13C HSQC correlation spectroscopy establishes that CH2D and CHD2 isotopomers are not produced with this method. This approach offers enhanced labeling of Leu methyl groups over previous methods that utilize Val as the labeling agent and is more cost effective.     

The effect of externally applied electrostatic fields,microwave radiation and electric currents on plants and other organisms,with special reference to weed control

A wide-ranging review is presented of the effects of various forms of externally applied electrical energy upon plants and other organisms. Although investigations involving both small and large amounts of energy directed at the targets are considered, a particular emphasis of this review is the feasibility of each type of electrical stimulation for weed control. Electrostatic fields ranging from 100 V m^?1 to 800 kV m^?1 have been applied to plants under laboratory conditions and in field trials since the 1880’s. Some beneficial effects have been reported (e.g. increase in yield from both cereal and vegetable crops), but the results have been erratic and the electrical conditions leading to definite benefits on a large scale could not be confidently predicted from early studies. High electric fields are reported to damage plants if currents greater than 10^?6 A are induced to flow through leaves causing corona discharges from the tips. The nature of the damage and the effects on metabolic processes are discussed. The results from experiments on the growth of plants in which the density and charge of air ions have been varied are also reviewed. The effects of microwave radiation (mostly 2450 MHz) upon seeds, plants and other organisms in soil are discussed. These effects depend upon the power density of the radiation and the electrical properties of the targets. Factors such as size of seeds and plants, shape and moisture content are important, as are the properties of the soil irradiated (notably water content). Although microwaves can be effective in killing plants and also seeds that are buried several centimeters deep in soil, high power equipment is required and treatment times are long e.g. a 60 kW machine could take up to 92.6 hours per hectare. Other experiments reported show that microwave radiation can kill nematodes in the soil and that it is also very effective in killing fungi and bacteria. The potential of the various possible uses of microwave radiation in agriculture is also described. Electric currents have been caused to flow through plants by the application of electrodes to the leaves. The effects range from nil, when 50–100 V and 1 or 2 μA are used, to very striking when voltages from 5 to 15 kV are applied causing currents of several amperes to flow and resulting in the rapid destruction of the target. Small electric currents passed through soil containing plants are reported to increase their growth. The effects of small current on the growth of individual leaves are reviewed. The use of high voltage tractor-borne equipment for weed control is also considered.