Molecular ecology and polyphasic characterization of the microbiota associated with semi-dry processed coffee (Coffea arabica L.)

This work was aimed at isolating and identifying the microbiota present during the semi-dry method of coffee processing using polyphasic methods and to evaluate microbial diversity with PCR-DGGE. Samples of Coffea arabica L. were collected during different processing stages in southern Minas Gerais, Brazil. The bacterial and fungal isolates were phenotypically characterised and grouped according to the ARDRA technique, in which the 16–23S and ITS1-5.8S regions of the rDNA were sequenced for species identification. The bacterial counts varied from 3.7 to 7 log CFU g⁻¹. The yeast counts ranged from 3.4 to 6.9 log CFU g⁻¹, and the filamentous fungal population varied from 2 to 3.7 log CFU g⁻¹. Bacillus subtilis, Escherichia coli, Enterobacter agglomerans, Bacillus cereus and Klebsiella pneumoniae were the predominant bacteria detected during the processing of the coffee, and Pichia anomala, Torulaspora delbrueckii and Rhodotorula mucilaginosa were the dominant yeasts. All of the yeast and bacterial species detected by PCR-DGGE were isolated using culture-dependent methods, with the exception of one uncultivable bacterial species. Aspergillus was the most common genus among the filamentous fungal isolates. The use of polyphasic methods allowed a better characterization of the microbiota that is naturally present in semi-dry processed coffee.     

Toxicity of Phenolic Compounds Extracted from Bark Residues of Different Ages

In Quebec, Canada, industrial bark wastelands cover several hundred hectares of land. Bark residue that has piled up for decades tends to remain free of vegetation for years. To assess the revegetation potential of such sites, we sought to determine those factors responsible for poor plant growth. Phenolic compounds from fresh to 20-year-old bark residues were extracted with four solvents and quantified by high-performance liquid chromatography (HPLC). We simulated solutions (mixtures of standard phenolic compounds) to evaluate the potential toxic effects of phenolic compounds on the rhizobial growth, germination index, plant growth, nodule number, and nitrogen fixation activity of two legume species under laboratory conditions. The concentration of individual phenolic compounds varied from none detected to 350 μg/g bark residue. The extracted phenolic compounds differed among solvents and bark residues. The highest concentration of total phenolic compounds was from fresh bark; most of these were soluble in water or 0.1 M NaOH. For older bark residues, the total phenolic content depended on solvent strength, generally in the order of 2.0 M NaOH > 0.1 M NaOH = hot water > cold water. The biological activity of the simulated bark extracts was not established with the rhizobial growth inhibition test but with the germination index and rhizobium–legume symbiosis tests. With these, the toxicity of the simulated phenolic extracts decreased from fresh to the older bark residues. Plant dry weight, nodule number, and nitrogen fixation activity of vetch (Vicia sativa L.) were less negatively affected by high concentrations of phenolics than birdsfoot trefoil (Lotus corniculatus L.), although birdsfoot trefoil grew at lower concentrations. The rhizobium–legume symbiosis has potential for revegetating bark wastelands with less than 1 year old and older bark residues.     

New Enantioselective Synthesis of Monofluorinated Pyridines Designed for the Preparation of Chemical Libraries

Chiral pyridines with a fluorine atom in the benzylic position are easily accessible from optically active propargylic fluorides by using the Bohlmann–Rahtz reaction. Such pyridines, possessing four points of molecular diversity, are useful scaffolds for the preparation of chemical libraries.     

Arsenic transformation and mobilization from minerals by the arsenite oxidizing strain WAO

Analysis of arsenic concentrations in New Jersey well water from the Newark Basin showed up to 15% of the wells exceed 10 microg L(-1), with a maximum of 215 microg L(-1). In some geologic settings in the basin, this mobile arsenic could be from the weathering of pyrite (FeS2) found in black shale that contains up to 4% arsenic by weight. We hypothesized that under oxic conditions at circumneutral pH, the microbially mediated oxidation of sulfide in the pyrite lattice would lead to the release of pyrite-bound arsenic. Moreover, the oxidation of aqueous As(III) to As(V) by aerobic microorganisms could further enhance arsenic mobilization from the solid phase. Enrichment cultures under aerobic, As(III)-oxidizing conditions were established under circumneutral pH with weathered black shale from the Newark Basin as the inoculum source. Strain WAO, an autotrophic inorganic-sulfur and As(III)-oxidizer, was isolated and phylogenetically and physiologically characterized. Arsenic mobilization studies from arsenopyrite (FeAsS) mineral, conducted with strain WAO at circumneutral pH, showed microbially enhanced mobilization of arsenic and complete oxidation of released arsenic and sulfur to stoichiometric amounts of arsenate and sulfate. In addition, WAO preferentially colonized pyrite on the surface of arsenic-bearing, black shale thick sections. These findings support the hypothesis that microorganisms can directly mobilize and transform arsenic bound in mineral form at circumneutral pH and suggest that the microbial mobilization of arsenic into groundwater may be important in other arsenic-impacted aquifers.     

Customer perception of switch-feel in luxury sports utility vehicles

Successful new product introduction requires that product characteristics relate to the customer on functional, emotional, aesthetic and cultural levels. As a part of research into automotive human machine interfaces (HMI), this paper describes holistic customer research carried out to investigate how the haptics of switches in luxury sports utility vehicles (SUVs) are perceived by customers. The application of these techniques, including an initial proposal for objective specifications, is addressed within the broader new product introduction context, and benefits described.One-hundred and one customers of SUVs assessed the feel of automotive push switches, completing the tasks both in, and out of vehicles to investigate the effect of context. Using the semantic differential technique, hedonic testing, and content analysis of customers’ verbatim comments, a holistic picture has been built up of what influences the haptic experience. It was found that customers were able to partially discriminate differences in switch-feel, alongside considerations of visual appearance, image, and usability. Three factors named ‘Affective’, ‘Robustness and Precision’, and ‘Silkiness’ explained 61% of the variance in a principle components analysis. Correlations of the factors with acceptance scores were 0.505, 0.371, and 0.168, respectively.     

Microstructure and nutrient composition of hairless canary seed and its potential as a blending flour for food use

Canary seed is a true cereal with unique composition. The current study employed light and fluorescence microscopy to visualise starch, protein, phenolics and phytate in hairless canary seed (CDC Maria), a cultivar developed potentially for food use. Macronutrients, minerals and vitamins were evaluated in the developed cultivar and compared with a commercial hairy canary seed, cv. Keet. A control common wheat, cv. Katepwa, was grown adjacent to the canary seed varieties. The compositions of the two canary seed varieties were found to be similar with an average of 55.8 g/100 g of starch, 23.7% g/100 g of protein, 7.9% of crude fat, 7.3 g/100 g of total dietary fibre, 1.8 g/100 g of soluble sugar and 2.3 g/100 g of total ash in the whole grain. Regardless of the milling fraction (whole grain flour, white flour or bran), canary seed had more protein and crude fat and less starch, total dietary fibre and soluble sugar than had wheat. It also had higher concentrations of several minerals and vitamins than did wheat. The structure of the canary seed grain exhibited compound starch granules and protein bodies embedded in a protein matrix similar to that of the oat kernel. Baking tests showed that bread made with 100% hairless canary seed flour was significantly lower in loaf volume and crust and crumb colour than was wheat bread. However, bread with loaf volume, specific volume and crust colour comparable to those of the bread control was achieved by using up to 25% of hairless canary seed or 15% of roasted canary seed flour, thus demonstrating its potential for food applications.     

The recognition of high molecular weight melanoidins as the main components responsible for radical-scavenging capacity of unheated and heat-treated Canadian honeys

As the Maillard reaction is known to occur in heat-treated foods, unheated and heated honey samples were subjected to the activity-guided fractionation and size-exclusion chromatography to compare the degree of browning, radical scavenging activity (ORAC) and molecular size of the fractions obtained. Heat-treatment increased browning in the fractions of light- and medium-coloured honeys (p < 0.002), accelerated the formation of high molecular weight melanoidins (85–232 kDa) and significantly increased ORAC values (p < 0.0001). In contrast, melanoidin content and ORAC decreased in the fractions of heat-treated dark buckwheat honey (p < 0.001). Unheated dark honey contained a significantly higher amount of melanoidins than other honeys (p < 0.007). Together, results showed that at low initial concentration of melanoidins, heat-treatment accelerated formation of new melanoidins and increased ORAC, while at high concentration it caused decrease and a reduction of radical scavenging activity. This study emphasises the importance of non-enzymatic browning in the prediction of the antioxidant pool in thermally processed honeys.     

Design,Synthesis, Radiosynthesis and Biological Evaluation of Fenretinide Analogues as Anticancer and Metabolic Syndrome-Preventive Agents

Fenretinide (4-HPR) is a synthetic derivative of all-trans-retinoic acid (ATRA) characterised by improved therapeutic properties and toxicological profile relative to ATRA. 4-HPR has been mostly investigated as an anti-cancer agent, but recent studies showed its promising therapeutic potential for preventing metabolic syndrome. Several biological targets are involved in 4-HPR's activity, leading to the potential use of this molecule for treating different pathologies. However, although 4-HPR displays quite well-understood multitarget promiscuity with regards to pharmacology, interpreting its precise physiological role remains challenging. In addition, despite promising results in vitro, the clinical efficacy of 4-HPR as a chemotherapeutic agent has not been satisfactory so far. Herein, we describe the preparation of a library of 4-HPR analogues, followed by the biological evaluation of their anti-cancer and anti-obesity/diabetic properties. The click-type analogue 3 b showed good capacity to reduce the amount of lipid accumulation in 3T3-L1 adipocytes during differentiation. Furthermore, it showed an IC₅₀ of 0.53±0.8 μM in cell viability tests on breast cancer cell line MCF-7, together with a good selectivity (SI=121) over noncancerous HEK293 cells. Thus, 3 b was selected as a potential PET tracer to study retinoids in vivo, and the radiosynthesis of [¹⁸F] 3b was successfully developed. Unfortunately, the stability of [¹⁸F] 3b turned out to be insufficient to pursue imaging studies.