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Selected AbstractsAmoebal pathogens as emerging causal agents of pneumoniaFEMS MICROBIOLOGY REVIEWS, Issue 3 2010Frédéric Lamoth Abstract Despite using modern microbiological diagnostic approaches, the aetiological agents of pneumonia remain unidentified in about 50% of cases. Some bacteria that grow poorly or not at all in axenic media used in routine clinical bacteriology laboratory but which can develop inside amoebae may be the agents of these lower respiratory tract infections (RTIs) of unexplained aetiology. Such amoebae-resisting bacteria, which coevolved with amoebae to resist their microbicidal machinery, may have developed virulence traits that help them survive within human macrophages, i.e. the first line of innate immune defence in the lung. We review here the current evidence for the emerging pathogenic role of various amoebae-resisting microorganisms as agents of RTIs in humans. Specifically, we discuss the emerging pathogenic roles of Legionella -like amoebal pathogens, novel Chlamydiae (Parachlamydia acanthamoebae, Simkania negevensis), waterborne mycobacteria and Bradyrhizobiaceae (Bosea and Afipia spp.). [source] Metabolic engineering of Saccharomyces cerevisiae for the synthesis of the wine-related antioxidant resveratrolFEMS YEAST RESEARCH, Issue 1 2003John V.W. Becker Abstract The stilbene resveratrol is a stress metabolite produced by Vitis vinifera grapevines during fungal infection, wounding or UV radiation. Resveratrol is synthesised particularly in the skins of grape berries and only trace amounts are present in the fruit flesh. Red wine contains a much higher resveratrol concentration than white wine, due to skin contact during fermentation. Apart from its antifungal characteristics, resveratrol has also been shown to have cancer chemopreventive activity and to reduce the risk of coronary heart disease. It acts as an antioxidant and anti-mutagen and has the ability to induce specific enzymes that metabolise carcinogenic substances. The objective of this pilot study was to investigate the feasibility of developing wine yeasts with the ability to produce resveratrol during fermentation in both red and white wines, thereby increasing the wholesomeness of the product. To achieve this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to be introduced to produce p -coumaroyl-CoA, one of the substrates required for resveratrol synthesis. The other substrate for resveratrol synthase, malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid biosynthesis. We hypothesised that production of p -coumaroyl-CoA and resveratrol can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216) from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in laboratory strains of S. cerevisiae. This yeast has the ability to metabolise p -coumaric acid, a substance already present in grape must. This compound was therefore added to the synthetic media used for the growth of laboratory cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained and tested for production of resveratrol. Following ,-glucosidase treatment of organic extracts for removal of glucose moieties that are typically bound to resveratrol, the results showed that the yeast transformants had produced the resveratrol ,-glucoside, piceid. This is the first report of the reconstruction of a biochemical pathway in a heterologous host to produce resveratrol. [source] Search for direct empirical spatial correlation signatures of the critical triggering earthquake modelGEOPHYSICAL JOURNAL INTERNATIONAL, Issue 3 2004G. Ouillon SUMMARY We propose a new test of the critical earthquake model based on the hypothesis that precursory earthquakes are ,actors' that create fluctuations in the stress field which exhibit an increasing correlation length as the critical large event becomes imminent. Our approach constitutes an attempt to build a more physically based time-dependent indicator (cumulative scalar stress function), in the spirit of, but improving on, the cumulative Benioff strain used in previous works documenting the phenomenon of accelerating seismicity. Using a simplified scalar space and time-dependent viscoelastic Green's function in a two-layer model of the Earth's lithosphere, we compute spatiotemporal pseudo-stress fluctuations induced by a series of events before four of the largest recent shocks in southern California. Through an appropriate spatial wavelet transform, we then estimate the contribution of each event in the series to the correlation properties of the simplified pseudo-stress field around the location of the mainshock at different scales. This allows us to define a cumulative scalar pseudo-stress function which reveals neither an acceleration of stress storage at the epicentre of the mainshock nor an increase of the spatial stress,stress correlation length similar to those observed previously for the cumulative Benioff strain. The earthquakes we studied are thus either simple ,witnesses' of a large-scale tectonic organization, or are simply unrelated, and/or the Green's function describing interactions between earthquakes has a significantly longer range than predicted for standard viscoelastic media used here. [source] Host-derived media used as a predictor for low abundant, in planta metabolite production from necrotrophic fungiJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2006D.P. Overy Abstract Aims:,Penicillium ser. Corymbifera strains were assayed on a variety of media and from infected Allium cepa tissues to evaluate the stimulation and in planta prediction of low abundance metabolites. Methods and Results:, Stimulated production of corymbiferones and the corymbiferan lactones were observed for Penicillium albocoremium, Penicillium allii, Penicillium hirsutum, Penicillium hordei and Penicillium venetum strains cultured on tissue media. Target metabolites were sporadically detected from strains cultured on common laboratory media (CYA, MEA and YES). Up to a 376 times increase in corymbiferone and corymbiferan lactone production was observed when culture extracts from CYA and A. cepa agar were compared by high pressure liquid chromatography with ultraviolet and mass spectrometry (LC-UV-MS). The novel metabolite corymbiferone B was purified and structure elucidated from a P. allii/A. cepa tissue medium extract. In planta expression of low abundance, target metabolites were confirmed from infected A. cepa tissue extracts by LC-UV-MS. Conclusions:, Secondary metabolite production was directly dependent and influenced by media conditions, resulting in the stimulated production of low abundance metabolites on host-derived media. Significance and Impact of the Study:, The use of macerated host tissue media can be applied in vitro to predict in planta expression of low abundance metabolites and aid in metabolite origin annotation during in planta metabolomic investigations at the host/pathogen interface. [source] Phytate degradation by micro-organisms in synthetic media and pea flourJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2002M. Fredrikson Aims: To screen micro-organisms for the ability to produce phytase enzyme(s) and to use promising strains for the fermentation of pea flour. Methods and Results: Two methods using the indirect estimation of phytate degradation were evaluated and both shown to be inadequate. A third method, measuring the inositol phosphate (IP3,IP6) content directly during fermentation, was used instead of the indirect estimations of phytate degradation. In synthetic media, some strains required customized conditions, with no accessible phosphorus sources other than phytate, to express phytase activity. The repression of phytase synthesis by inorganic phosphorus was not detected during fermentation with pea flour as substrate and seemed to be less significant with a higher composition complexity of the substrate. None of the tested lactic acid bacteria strains showed phytase activity. Conclusions: The methodology for the phytase screening procedure was shown to be critical. Some of the screening methods and media used in previous publications were found to be inadequate. Significance and Impact of the Study: This paper highlights the pitfalls and difficulties in the evaluation of phytase production by micro-organisms. The study is of great importance for future studies in this area. [source] Comparison of ATP and in vivo bioluminescence for assessing the efficiency of immunomagnetic sorbents for live Escherichia coli O157:H7 cellsJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2002W. Sun Aims:,To develop methods to assess the efficiency of immunomagnetic separation (IMS). Methods and Results:,The capturing efficiency of biosorbents for Escherichia coli O157:H7, constructed using streptavidin-coated magnetic beads and biotinylated antibodies, was tested using both in vivo and ATP bioluminescence. Both methods were suitable for the enumeration of bacteria captured by the biosorbents. The level of both ATP and in vivo bioluminescence depended on the media used, but was unaffected by the magnetic beads. The capture efficiency depended on time and sample volume, but did not depend on the length of spacer arm of the biotinylation agent. For cell concentrations of , 105 cfu ml,1, in a 1-ml sample volume, nearly 80,85% recovery of the pathogen was observed after 0·5 h of incubation. For an 11-ml sample containing 104 cfu ml,1, maximum recovery (50% of cells) was achieved only after 2 h incubation. Conclusions:,The detection limit of an ATP-based bioluminescent assay for E. coli O157:H7 was reduced by 1 log cycle after optimization of IMS. The bioluminescent methods could be used for screening and testing the affinity of antibodies or other affinity elements of biosorbents towards live bacterial cells. Significance and Impact of the Study:,Bioluminescent assays provide an easy way to optimize conditions for the capture of bacteria by biosorbents in real time. [source] Comparison of three enrichment media for the isolation of Campylobacter spp. from foodsJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2000C.L. Baylis Aim: This study compared the performance of three Campylobacter enrichment broths: Bolton broth (BB), Campylobacter Enrichment broth (CEB) and Preston broth (PB). Methods and Results: Pure cultures of target and competitor organisms, and naturally-contaminated food samples, were used to establish the performance of these media. In pure culture the PB supported the growth of the greatest number of strains of Campylobacter spp. but failed to inhibit some competitor organisms. The CEB showed the opposite result, inhibiting all 15 competitor organisms used but failing to support the growth of five Campylobacter strains. By comparison, BB showed the best compromise between inhibition of competitors and growth of Campylobacter. Conclusions: Plates inoculated with BB and CEB food enrichments resulted in more Campylobacter growth than those inoculated with PB, which supported significantly less typical growth (P , 0·001). The most common competitor organism isolated from PB was Escherichia coli, and Pseudomonas spp. were frequently isolated from BB and CEB. Both BB and CEB were better than PB for the isolation of Campylobacter from naturally-contaminated foods, although BB yielded more confirmed Campylobacter growth than CEB. Significance and Impact of the Study: This study highlighted differences in performance of media used to isolate Campylobacter spp. from foods. [source] Biological, pharmaceutical, and analytical considerations with respect to the transport media used in the absorption screening system, Caco-2JOURNAL OF PHARMACEUTICAL SCIENCES, Issue 8 2003Françoise M. Ingels Abstract During the evaluation and selection of drug candidates, the Caco-2 cell culture system is commonly used for the determination of intestinal transport characteristics and to anticipate permeability limited drug absorption. Although classic HBSS-like buffered salt solutions are commonly used to perform Caco-2 transport experiments, different shortcomings (e.g., adsorption and low solubility) have been associated with the use of plain aqueous buffers. As transport experiments performed with unoptimized conditions may compromize the value of the Caco-2 model as a permeation screening tool, many efforts have been made to optimize the experimental conditions of Caco-2 transport assays. In this minireview, the hurdles associated with the use of saline aqueous buffers in Caco-2 transport experiments are summarized and the different options, which have been proposed to overcome these issues, are reviewed and discussed. Biologically, pharmaceutically, as well as analytically relevant media affecting the outcome of the transport experiments are described. Unfortunately, up to now, no systematic studies comparing the different experimental conditions have been performed, jeopardizing the possibility to define a (single) optimal solution to overcome the different issues associated with the use of saline aqueous buffers. Based on the reported options it can be proposed to use DMSO (,1%) in standard screening procedures for the ranking of compounds based on their apical to basolateral transport. If compounds are not soluble in DMSO 1%, dimethylacetamide (3%) or N -1-methyl-pyrrolidone (2.5%) are good alternatives. However, these options do not imitate the in vivo situation. If one wants to take into account the physiological relevance of the media, the use of a biologically relevant apical medium (e.g., FASSIF) in combination with an analytically friendly, sink condition creating basolateral solvent (e.g., containing a micelle forming agent) can be suggested. © 2003 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 92:1545,1558, 2003 [source] The effect of culture preservation techniques on patulin and citrinin production by Penicillium expansum LinkLETTERS IN APPLIED MICROBIOLOGY, Issue 4 2002I.M. Santos Aims: To study the influence of culture preservation methods and culture conditions on the production of the mycotoxins patulin and citrinin by Penicillium expansum. Methods and results: Ten strains of Penicillium expansum were preserved using subculture and maintenance at 4 °C, mineral oil, drying on silica gel and freeze-drying. Patulin and citrinin production was assessed on yeast extract sucrose agar (YES) and grape juice agar (GJ), using TLC before and after 0·5, 2,3, 6 and 12 months preservation. Citrinin was detected in all cultures for all preservation techniques on YES. The patulin profiles obtained differed with strain and culture media used. Conclusions: Citrinin production seems to be a stable character for the tested strains. There is a tendency for patulin detection with time apparently more consistent for silica gel storage and freeze-drying, especially when the strains are grown on GJ. Significance and Impact of the Study: Variability in the profiles of the mycotoxins tested seems to be more strain-specific than dependent on the preservation technique used. [source] | ||||||||||||||||||||||