Trend reversal of nitrate in Danish groundwater--a reflection of agricultural practices and nitrogen surpluses since 1950

This paper assesses the long-term development in the oxic groundwater nitrate concentration and nitrogen (N) loss due to intensive farming in Denmark. First, up to 20-year time-series from the national groundwater monitoring network enable a statistically systematic analysis of distribution, trends, and trend reversals in the groundwater nitrate concentration. Second, knowledge about the N surplus in Danish agriculture since 1950 is used as an indicator of the potential loss of N. Third, groundwater recharge CFC (chlorofluorocarbon) age determination allows linking of the first two data sets. The development in the nitrate concentration of oxic groundwater clearly mirrors the development in the national agricultural N surplus, and a corresponding trend reversal is found in groundwater. Regulation and technical improvements in the intensive farming in Denmark have succeeded in decreasing the N surplus by 40% since the mid 1980s, while at the same time maintaining crop yields and increasing the animal production of especially pigs. Trend analyses prove that the youngest (0-15 years old) oxic groundwater shows more pronounced significant downward nitrate trends (44%) than the oldest (25-50 years old) oxic groundwater (9%). This amounts to clear evidence of the effect of reduced nitrate leaching on groundwater nitrate concentrations in Denmark.     

Development and application of new nucleic acid-based technologies for microbial community analyses in foods

Several challenges still persist in the analysis of microorganisms in foods, particularly in studies of complex communities. Nucleic acid-based methods are promising tools in addressing new questions concerning microbial communities. We have developed several new methods in the field of nucleic acid-based microbial community analyses. These methods cover both sample preparation and detection approaches. The sample preparation method involves simplified DNA purification using paramagnetic beads. As an extension of this method, the same paramagnetic beads are used for both cell separation and DNA purification. This enables full automation. The separate detection of viable and dead bacteria is a major issue in nucleic acid-based diagnostics. We have applied a living/dead dye that binds covalently to DNA and inhibits the PCR from dead cells. In addition, a DNA array-based detection assay has been developed. The assay combines the specificity obtained by enzymatic labeling of DNA probes with the possibility of detecting several targets simultaneously by DNA array hybridization. In combination with 16S rDNA amplification, this is a promising tool for community analyses. Also, we have developed a novel approach for multiplex quantitative PCR. The multiplex PCR has been combined with our DNA array-based detection method. Finally, we are now in the process of adapting a system for monitoring microbial growth and death in real-time through the tagging of bacteria with green fluorescent protein (GFP) combined with fluorescence detection using a high-resolution confocal laser scanner.     

Biofeedback and relaxation therapy for chronic temporomandibular joint pain: Predicting successful outcomes.

57 patients with chronic temporomandibular joint (TMJ) pain were randomly assigned to receive either relaxation or biofeedback therapy. 27 Ss (mean age 35.6 yrs) listened to tape-recorded relaxation once a week at the therapist's office for 3 sessions and were encouraged to practice daily. 30 Ss (mean age 43 yrs) participated in biofeedback sessions and were instructed to practice relaxation for 20 min/day between sessions. Results show no significant differences in outcomes. However, successful Ss in the 2 conditions differed from each other. Successful relaxation Ss tended to be younger, had TMJ pain for a shorter period of time, and reported problems with other psychophysiologic disorders. Successful biofeedback Ss were older, married, had TMJ pain for a longer period of time, and had not received prior equilibration treatment. Equilibration and presence of other disorders were related to both short- and long-term outcomes, suggesting they may be useful as predictors of outcome. It is also suggested that knowledge of pretherapy factors may allow for optimal assignment to therapy conditions. (30 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Partial purification and characterization of free and immobilized lipases fromMucor miehei

An extracellular lipase, a glycoprotein, produced by fermentation with a selected strain ofMucor miehei has been partially purified in two forms, A and B. The two forms have a high degree of antigenic identity and have similar pH-activity profiles with tributyroylglycerol as a substrate with optima at pH 7. They differ as follows: A, in contrast to B, requires activation at alkaline pH before analysis; A binds with concanavalin-A more completely than B; the net charges are slightly different at pH 8; and the isoelectric points are different. Our results indicate that the B lipase is formed by partial deglycosylation of the A lipase and that this influences the activity toward emulsions. In addition, the two enzymes have been immobilized by adsorption. These preparations and the soluble forms were highly specific for primary esters of triacylglycerols (TG); they usually hydrolyzed TG of 12∶0, 14∶0, 16∶0, and 18∶1 more rapidly than those of 4∶0, 6∶0, and 8∶0 and 10∶0 in mixtures of monoacid TG (4∶0 to 18∶1); and they were not stereospecific for TG. Immobilization altered the specificity of the preparations somewhat, in that slightly more 14∶0 and 16∶0 were released. Scientific Contribution No. 1192, Storrs Agricultural Experiment Station, University of Connecticut, Storrs, Connecticut.     

4,4‐Dimethyl‐ and Diastereomeric 4‐Hydroxy‐4‐methyl‐ (2S)‐Glutamate Analogues Display Distinct Pharmacological Profiles at Ionotropic Glutamate Receptors and Excitatory Amino Acid Transporters

Subtype‐selective ligands are of great interest to the scientific community, as they provide a tool for investigating the function of one receptor or transporter subtype when functioning in its native environment. Several 4‐substituted (S)‐glutamate (Glu) analogues were synthesized, and altogether this approach has provided important insight into the structure–activity relationships (SAR) for ionotropic and metabotropic glutamate receptors (iGluRs and mGluRs), as well as the excitatory amino acid transporters (EAATs). In this work, three 4,4‐disubstituted Glu analogues 1 – 3 , which are hybrid structures of important 4‐substituted Glu analogues 4 – 8 , were investigated at iGluRs and EAATs. Collectively, their pharmacological profiles add new and valuable information to the SAR for the iGluRs and EAAT1–3.     

Biotransformation of hop derived compounds by Brettanomyces yeast strains

Several hop derived compounds in wort are known to be converted by yeast during fermentation, influencing the overall perception of the beer. A deeper understanding of such metabolic processes during fermentation is needed to achieve better control of the outcome. Here, the interaction between hop derived compounds and the yeast genera Brettanomyces was studied. Several Brettanomyces strains with different genomic backgrounds were selected, focusing on two traits: beta-glucosidase activity and nitrate assimilation. The role of three beta-glucosidases present in Brettanomyces bruxellensis and Brettanomyces anomalus and their impact on the final monoterpene alcohol profile was analysed. The beta-glucosidase activity was highly strain dependent, with B. anomalus CRL-49 exhibiting the highest conversion. Such activity could not be related to the release of aglycones from hops during fermentation, as a substantial part of such activity was intracellular. Nevertheless, the reduction of geraniol to β-citronellol was remarkably efficient for all Brettanomyces strains during fermentation, and it is suggested that two oxidoreductases BbHye2 and BbHye3 may have an influence. Moreover, the transfer of nitrate from hops to wort and its further assimilation by Brettanomyces species was analysed. The amount of nitrate in wort proved to be linearly proportional to the contact time of the hops with the wort. The level of nitrate assimilation by yeast was shown to be dependent on the nitrate assimilation cluster (YNR, YNI, YNT). Hence, the desired yeast strains may be selected according to the genetic make-up. © 2020 The Authors. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

‘Salesman? Hell no!’ Identity struggles of nascent design entrepreneurs

The present study offers new input to the discussion of how entrepreneurship education and training programmes can be tailored to suit design professionals. To a large extent, existing entrepreneurship teaching for creative people is based on a traditional administrative management logic that often results in a clash between entrepreneurial demands and creative identities. The paper is based on the following rationale: the better we understand designers' reasoning and their struggles concerning becoming entrepreneurs, the better we are able to design entrepreneurship learning experiences that meet their needs. Since designers' career‐making tends to be highly driven by their strong sense of identity, the paper takes an identity perspective. The empirical foundation of the research is based on observation studies and phenomenological interviews conducted during an eight‐week entrepreneurship training program. Twenty‐five nascent design entrepreneurs with a professional background as designers participated in the voluntary programme. The paper offers novel and critical insights into designers' experiences of the entrepreneurial identity and reasoning as they participate in entrepreneurship training.     

Real-time PCR quantification of the AM-toxin gene and HPLC qualification of toxigenic metabolites from Alternaria species from apples

Some Alternaria species are able to produce plant pathogenic as well as toxic metabolites. In both agriculture and the food industry it is important know if toxigenic Alternaria are present to rapidly employ the correct corrective actions. The purpose of this work was to establish a real-time PCR method, which can detect and quantify apple pathogenic and toxigenic Alternaria. An AM-toxin I primer set, which could recognize Alternaria DNA only, was designed by using primers complementary to the AM-toxin I gene. The method could detect small amounts of DNA (4 pg) and still obtain a large dynamic range (4 decades) without interference from apple material. Eight Alternaria isolates were analyzed for the presence of AM-toxin I gene and their production of secondary metabolites. Then analyses showed that all eight isolates contained the AM toxin gene and were able to produce the plant pathogenic tentoxin in addition to AM toxin I. The analyses also showed the production of tenuazonic acid, alternariols, Altenuene, altenusin and/or altertoxin I in pure culture. Analyses of inoculated apples showed that both the AM-toxin gene and alternariol monomethyl ether could be detected. Morphological analyses suggested that the eight Alternaria strains, though they all carried the AM toxin genes, probably belong to different but closely related un-described Alternaria taxa in the A. tenuissima species-group based on morphological and chemical differences.