Thermal resistance of different life stages of codling moth (Lepidoptera: Tortricidae)

Phytosanitation regulations in several international markets require postharvest treatments to control codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), in various commodities. Thermal treatments are gaining acceptance to replace chemical fumigation. Determining the most thermal-tolerant life stage is essential in the development of effective postharvest insect control protocols based on thermal energy. A heating block system was used to evaluate relative heat resistance of five different life stages of codling moth: white-ring eggs, black-head eggs, third-instar, fifth-instar, and diapausing larvae, at a heating rate of 15°C/min. The fifth-instar was the most heat-resistant life stage in the tested temperature range of 50–52°C except for diapausing larvae. Thermal death kinetic data of diapausing fifth-instar larvae were determined and also compared with the published TDT curve of non-diapausing larvae using the same heating block system. Both diapausing and non-diapausing larvae were dead after treatments at 50°C for 5 min and 52°C for 2 min. However, at the lower temperatures or shorter times, diapausing larvae had lower mortality than non-diapausing larvae.     

Effect of salt on structure-function relationships of cheese

Our objective was to determine the effect of salt on structural and functional properties of cheese. Unsalted Muenster cheese was obtained on 1 d, vacuum packaged, and stored for 10 d at 4 degrees C. The cheese was then cut into blocks that were vacuum packaged. After 4 d of storage at 4 degrees C, cheese blocks were high-pressure injected one, three, or five times, with a 20% (wt/wt) sodium chloride solution. Successive injections were performed 24 h apart. After 40 d of storage at 4 degrees C, cheese blocks were analyzed for chemical, structural, and functional attributes. Injecting sodium chloride increased the salt content of cheese, from 0.1% in the control, uninjected cheese to 2.7% after five injections. At the highest levels, salt injection promoted syneresis, and, after five injections, the moisture content of cheese decreased from 41 to 38%. However, the increased salt content caused a net weight gain. Cheese pH, soluble nitrogen, and total and soluble calcium content were unaffected. Cheese injected five times had a 4% increased area of cheese occupied by protein matrix compared with uninjected cheese. Hardness, adhesiveness, and initial rate of cheese flow increased, and cohesiveness decreased upon salt injection. However, the final extent of cheese flow, or melting was unaffected. We concluded that adding salt to cheese alters protein interactions, such that the protein matrix becomes more hydrated and expands. However, increasing the salt content of cheese did not cause an exchange of calcium with sodium. Therefore, calcium-mediated protein interactions remain a major factor controlling cheese functionality.     

The pYC plasmids, a series of cassette-based yeast plasmid vectors providing means of counter-selection

A series of 24 general-purpose yeast plasmid vectors has been constructed. The plasmid series is composed of inter-replaceable cassettes, allowing for easy interconversion of plasmid types. In addition to the usual replication origins, selectable markers and multiple cloning sites (MCS), cassettes dedicated to counter-selection have been constructed. A pair of unique 8 bp restriction enzyme recognition sites flank each type of cassette, FseI in the case of yeast replication origins, AscI in the case of selectable markers, PacI in the case of counter-selectable markers and NotI in the case of the MCS. Thus, any given cassette can be replaced by another cassette of the same type, facilitating interconversion of any given plasmid from one type to another, even after the insertion of DNA into the MCS. Hence, the plasmids have been named pYC for 'yeast cassettes'. The cassettes consist of either NONE, CEN4/ARS or 2micro as replication origin, either URA3, MET2-CA (Lg-MET2) or the G418 resistance gene (the apt1 gene from bacterial transposon Tn903, encoding aminoglycoside phosphotransferase) as selectable markers, either NONE, PMET25-PKA3 or PCHA1-PKA3 as counter-selectable marker, and the MCS, containing recognition sites for AflII, AvrII, BspEI, PmeI, SacII, SalI, SunI, BamHI, EcoRI, HindIII, KpnI, MluI, NarI and SacI (of which the seven first are unique in all plasmids). The counter-selectable markers consist of the PKA3 gene under control of the conditional MET25 or CHA1 promoters. At activating conditions these promoters express the PKA3 gene at toxic levels, facilitating easy selection for loss of plasmid or 'loop-out' of plasmid DNA sequence after genomic integration.     

Precision Glass Molding: Validation of an FE Model for Thermo-Mechanical Simulation

In precision glass molding process, the required accuracy for the final size and shape of the molded lenses as well as the complexity of this technology calls for a numerical simulation. The current paper addresses the development of an FE model for thermo-mechanical simulation of the precision glass molding process including heating, pressing, and cooling stages. Temperature-dependent viscoelastic and structural relaxation behavior of the glass material are implemented through a FORTRAN material subroutine (UMAT) into the commercial FEM program ABAQUS, and the FE model is validated with a sandwich seal test. Subsequently, precision molding of several glass rings is performed at three different pressing temperatures, and the experimental deformation of the glass rings at the end of the molding is compared with the predicted ones from FE simulation. Furthermore, the transient and residual stress distribution inside the glass rings are calculated by the developed FE model, and the effects of some important process parameters such as interface friction and mold temperature on the FE results are assessed. The developed FE model can be employed to predict the deformation behavior, final size/shape, and the residual stress state inside the glass lenses in a precision glass molding process.     

辣椒碱及衍生物的研究进展

目的：为获得较为理想的镇痛药进行辣椒碱及类似物的进一步修饰改造提供依据。方法：通过对国内外文献中辣椒碱合成路线的归纳、比较．整理出一条较好的工艺路线,同时也对辣椒碱结构改造及修饰物进行构效关系进行比较。结果：只有保留了香革胺或其电子等排体部分的化合物具有镇痛作用,苯环不同取代基不同位置的取代及侧链的改变都对药效及刺激性有一定影响。结论：侧链和香草胺被其不同生物电子等排取代可能是进行辣椒碱及类似物结构修饰改造的方向。     

Minimal substrate features for Erm methyltransferases defined by using a combinatorial oligonucleotide library

Erm methyltransferases are prevalent in pathogenic bacteria and confer resistance to macrolide, lincosamide, and streptogramin B antibiotics by specifically methylating the 23S ribosomal RNA at nucleotide A2058. We have identified motifs within the rRNA substrate that are required for methylation by Erm. Substrate molecules were constructed in a combinatorial manner from two separate sets (top and bottom strands) of short RNA sequences. Modifications, including LNA monomers with locked sugar residues, were incorporated into the substrates to stabilize their structures. In functional substrates, the A2058 methylation target (on the 13- to 19-nucleotide top strand) was displayed in an unpaired sequence immediately following a conserved irregular helix, and these are the specific structural features recognized by Erm. Erm methylation was enhanced by stabilizing the top-strand conformation with an LNA residue at G2056. The bottom strand (nine to 19 nucleotides in length) was required for methylation and was still functional after extensive modification, including substitution with a DNA sequence. Although it remains possible that Erm makes some unspecific contact with the bottom strand, the main role played by the bottom strand appears to be in maintaining the conformation of the top strand. The addition of multiple LNA residues to the top strand impeded methylation; this indicates that the RNA substrate requires a certain amount of flexibility for accommodation into the active site of Erm. The combinatorial approach for identifying small but functional RNA substrates is a step towards making RNA-Erm complexes suitable for cocrystal determination, and for designing molecules that might block the substrate-recognition site of the enzyme.     

Formation of cereulide and enterotoxins by Bacillus cereus in fermented African locust beans

Afitin, iru and sonru are three spontaneously fermented African locust bean Benin condiments. The fermentation processes are exothermic, with temperatures mostly being above 40 °C. A total of 19 predominant Bacillus cereus isolates from afitin, iru and sonru, were investigated. The enterotoxin genes nhe (A, B, C) were present in all 19 isolates, the hbl (A, C, D) in one (afitin), and the cytK gene in three isolates (afitin). Levels of cytotoxicity to Vero cells and NheA production in BHI-broth was within the range of known diarrheal outbreak strains. Autoclaved cooked African locust beans inoculated with emetic (cereulide producing) B. cereus Ba18H2/RIF supported growth at 25, 30 and 40 °C with highly different maximum cereulide productions of 6 ± 5, 97 ± 3 and 0.04 ± 0.02 μg/g beans, respectively (48 h). For non-autoclaved cooked beans inoculated with 2, 4 and 6 log₁₀B. cereus Ba18H2/RIF spores/g beans, cereulide production was 5 ± 4, 64 ± 8 and 69 ± 34 μg/g beans, respectively at 24 h, while it was 70 ± 43, 92 ± 53 and 99 ± 31 μg/g at 48 h of fermentation at 30 °C. Even though high toxin levels were observed, to date there are no known reports on diarrhea or vomiting due to the consumption or afitin, iru and sonru in Benin, which also according to the present study is likely to be expected from the low levels of cereulide produced at 40 °C.     

Characterization of marama bean (Tylosema esculentum) by comparative spectroscopy: NMR, FT-Raman, FT-IR and NIR

The marama bean from Southern Africa has proven to be a source for production of various healthy food products. In order to exploit its commercial potential, it is important to know its chemical composition in more detail. In this study, marama beans from different geographical sites and harvest years were analyzed by use of infrared, near infrared, Raman, and ¹H as well as ¹³C nuclear magnetic resonance spectroscopy. These techniques can measure single beans in a rapid and non-destructive manner. By comparative application, the qualitative composition of the marama bean was explored in detail, revealing large amounts of protein, dietary fiber and unsaturated fat. The carbohydrate fraction was largely present as pectins and a minor fraction of smaller water soluble carbohydrates were tentatively assigned to raffinose. It is characteristic that the beans do not contain starch or ??-glucans and that the water soluble part of the proteins/peptides have a high content of the aromatic amino acid tyrosine.     

Analysis of secondary plant metabolites by indirect desorption electrospray ionization imaging mass spectrometry

Secondary metabolites in plant material can be imaged in a simple and robust way by creating an imprint of the plant material on a porous Teflon surface. The Teflon surface serves to extract compounds from the plant material for enhanced desorption electrospray ionization imaging analysis, while maintaining the spatial information of the sample. The method, which remedies for limitations in mass spectrometry imaging of compounds embedded in plant material, was demonstrated on leaves and petals of Hypericum perforatum and leaves of Datura stramonium.