A soil-based microbial biofilm exposed to 2,4-D: bacterial community development and establishment of conjugative plasmid pJP4

A soil suspension was used as a source to initiate the development of microbial communities in flow cells irrigated with 2,4-dichlorophenoxyacetic acid (2,4-D) (25 mu g ml(-1)). Culturable bacterial members of the community were identified by 16S rRNA gene sequencing and found to be members of the genera Pseudomonas, Burkholderia, Collimonas and Rhodococcus. A 2,4-D degrading donor strain, Pseudomonas putida SM 1443 (pJP4::gfp), was inoculated into flow cell chambers containing 2-day old biofilm communities. Transfer of pJP4::gfp from the donor to the bacterial community was detectable as GFP fluorescing cells and images were captured using confocal scanning laser microscopy (GFP fluorescence was repressed in the donor due to the presence of a chromosomally located lacl(q) repressor gene). Approximately 5-10 transconjugant microcolonies, 20-40 mu m in diameter, could be seen to develop in each chamber. A 2,4-D degrading transconjugant strain was isolated from the flow cell system belonging to the genus Burkholderia.

Matrix effects: The Achilles heel of quantitative high-performance liquid chromatography-electrospray-tandem mass spectrometry

High-performance liquid chromatography coupled by an electrospray ion source to a tandem mass spectrometer (HPLC-EST-MS/ MS) is the current analytical method of choice for quantitation of analytes in biological matrices. With HPLC-ESI-MS/MS having the characteristics of high selectivity, sensitivity, and throughput, this technology is being increasingly used in the clinical laboratory. An important issue to be addressed in method development, validation, and routine use of HPLC-ESI-MS/MS is matrix effects. Matrix effects are the alteration of ionization efficiency by the presence of coeluting substances. These effects are unseen in the chromatograrn but have deleterious impact on methods accuracy and sensitivity. The two common ways to assess matrix effects are either by the postextraction addition method or the postcolumn infusion method. To remove or minimize matrix effects, modification to the sample extraction methodology and improved chromatographic separation must be performed. These two parameters are linked together and form the basis of developing a successful and robust quantitative HPLC-EST-MS/MS method. Due to the heterogenous nature of the population being studied, the variability of a method must be assessed in samples taken from a variety of subjects. In this paper, the major aspects of matrix effects are discussed with an approach to address matrix effects during method validation proposed. (c) 2004 The Canadian Society of Clinical Chemists. All rights reserved.

Glucose affects monocarboxylate cotransporter (MCT) 1 expression during mouse preimplantation development

Cleavage-stage embryos have an absolute requirement for pyruvate and lactate, but as the morula compacts, it switches to glucose as the preferred energy source to fuel glycolysis. Substrates such as glucose, amino acids, and lactate are moved into and out of cells by facilitated diffusion. in the case of lactate and pyruvate, this occurs via H+-monocarboxylate cotransporter (MCT) proteins. To clarify the role of MCT in development, transport characteristics for DL-lactate were examined, as were mRNA expression and protein localisation for MCT1 and MCT3, using confocal laser scanning immunofluorescence in freshly collected and cultured embryos. Blastocysts demonstrated significantly higher affinity for DL-lactate than zygotes (K-m 20 +/- 10 vs 87 +/- 35 mmol lactate/l; P = 0.03 by linear regression) but was similar for all stages. For embryos derived in vivo and those cultured with glucose, MCT1 mRNA was present throughout preimplantation development, protein immunoreactivity appearing diffuse throughout the cytoplasm with brightest intensity in the outer cortical region of blastomeres. in expanding blastocysts, MCT1 became more prominent in the cytoplasmic cortex of blastomeres, with brightest intensity in the polar trophectoderm. Without glucose, MCT1 mRNA was not expressed, and immunoreactivity dramatically reduced in intensity as morulae died. MCT3 mRNA and immunoreactivity were not detected in early embryos. The differential expression of MCT1 in the presence or absence of glucose demonstrates that it is important in the critical regulation of pH and monocarboxylate transport during preimplantation development, and implies a role for glucose in the control of MCT1, but not MCT3, expression.

Resistance to persuasive messages as a function of majority and minority source status

Three experiments examined the extent to which attitudes following majority and minority influence are resistant to counter-persuasion. In Experiment 1, participants' attitudes were measured after being exposed to two messages which argued opposite positions (initial pro-attitudinal message and subsequent, counter-attitudinal counter-message). Attitudes following minority endorsement of the initial message were more resistant to a (second) counter-message than attitudes following majority endorsement of the initial message. Experiment 2 replicated this finding when the message direction was reversed (counter-attitudinal initial message and pro-attitudinal counter-message) and showed that the level of message elaboration mediated the amount of attitude resistance. Experiment 3 included conditions where participants received only the counter-message and showed that minority-source participants had resisted the second message (counter-message) rather than being influenced by it. These results show that minority influence induces systematic processing of its arguments which leads to attitudes which are resistant to counter-persuasion. (C) 2003 Elsevier Science (USA). All rights reserved.

Pacific pandanus fruit: an ethnographic approach to understanding an overlooked source of provitamin A carotenoids

Commonly recommended plant sources of provitamin A, such as dark green leafy vegetables, are not acceptable in many population groups. The objective of this study was to identify other indigenous foods that may be effectively promoted to alleviate vitamin A deficiency (VAD) and to gather information relevant to identification, production, acquisition, and consumption of foods relevant to a food-based VAD prevention strategy in the Federated States of Micronesia. An ethnographic study on edible pandanus cultivars, involving key informant interviews and observation was carried out. Analyses revealed a great range in carotenoid content. Several orange-coloured pandanus cultivars, all highly acceptable, contained high levels of carotenoid, almost meeting daily requirements in usual consumption patterns, whereas light yellow-coloured cultivars contained low levels. Availability has decreased substantially in recent years due to increased consumption of imported foods and general neglect of indigenous foods. High-carotenoid pandanus should be promoted for general enjoyment and health benefits.

Larval dispersal reveals regional sources and sinks in the Great Barrier Reef

We analysed simulated connectivity patterns for reef fish larvae in the Cairns section of the Great Barrier Reef, and identified 2 key subregions that exhibit regional scale source–sink dynamics. The source and sink were separated latitudinally by a boundary at 16.1°S, with the source subregion lying to the north. Larval transport between the 2 subregions was predominantly unidirectional, from north to south. Only a few local populations, described here as ‘gateway reefs’, were able to transport larvae from the sink subregion to the source subregion and thus maintain the connectedness of the metapopulation. The northern subregion was able to persist without external larval supply, but when conditions were recruitment limited, the southern subregion depended on larval supply from the north to persist. The relative autonomy of the northern subregion, and its importance in sustaining the southern subregion, will influence the effectiveness of conservation efforts.

Susceptibility of source plants to Sugarcane Fiji disease virus influences the acquisition and transmission of the virus by the planthopper vector Perkinsiella saccharicida

Fiji leaf gall (FLG) caused by Sugarcane Fiji disease virus (SCFDV) is transmitted by the planthopper Perkinsiella saccharicida. FLG is managed through the identification and exploitation of plant resistance. The glasshouse-based resistance screening produced inconsistent transmission results and the factors responsible for that are not known. A series of glasshouse trials conducted over a 2-year period was compared to identify the factors responsible for the erratic transmission results. SCFDV transmission was greater when the virus was acquired by the vector from a cultivar that was susceptible to the virus than when the virus was acquired from a resistant cultivar. Virus acquisition by the vector was also greater when the vector was exposed to the susceptible cultivars than when exposed to the resistant cultivar. Results suggest that the variation in transmission levels is due to variation in susceptibility of sugarcane cultivars to SCFDV used for virus acquisition by the vector.