Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

Characterization and Reconstitution of Human Lipoyl Synthase (LIAS) Supports ISCA2 and ISCU as Primary Cluster Donors and an Ordered Mechanism of Cluster Assembly

Lipoyl synthase (LIAS) is an iron–sulfur cluster protein and a member of the radical S-adenosylmethionine (SAM) superfamily that catalyzes the final step of lipoic acid biosynthesis. The enzyme contains two [4Fe–4S] centers (reducing and auxiliary clusters) that promote radical formation and sulfur transfer, respectively. Most information concerning LIAS and its mechanism has been determined from prokaryotic enzymes. Herein, we detail the expression, isolation, and characterization of human LIAS, its reactivity, and evaluation of natural iron–sulfur (Fe–S) cluster reconstitution mechanisms. Cluster donation by a number of possible cluster donor proteins and heterodimeric complexes has been evaluated. [2Fe–2S]-cluster-bound forms of human ISCU and ISCA2 were found capable of reconstituting human LIAS, such that complete product turnover was enabled for LIAS, as monitored via a liquid chromatography–mass spectrometry (LC–MS) assay. Electron paramagnetic resonance (EPR) studies of native LIAS and substituted derivatives that lacked the ability to bind one or the other of LIAS’s two [4Fe–4S] clusters revealed a likely order of cluster addition, with the auxiliary cluster preceding the reducing [4Fe–4S] center. These results detail the trafficking of Fe–S clusters in human cells and highlight differences with respect to bacterial LIAS analogs. Likely in vivo Fe–S cluster donors to LIAS are identified, with possible connections to human disease states, and a mechanistic ordering of [4Fe–4S] cluster reconstitution is evident.     

不同地区不同等级复烤烟叶中重要致香物质的分析比较

本文采用同时蒸馏萃取(SDE)/气相色谱/质谱联用法(GC/MS)对相同品种、不同地区、不同等级复烤烟叶中的重要致香物质进行分析比较;并且对同时蒸馏萃取法进行重复性和回收率试验。结果表明相同品种、不同地区、不同等级复烤烟叶中重要致香物质的含量差别较大;同时蒸馏萃取法具有良好的重复性和较高的回收率。     

The historical workstation project

Since 1978 research in the development of software dedicated to the specific problems of historical research has been undertaken at the Max-Planck-Institute für Geschichte in Göttingen. From a background of practical experiences during these years, a concept of what an appropriate workstation for an historian would be has been derived. It stresses the necessity of three components: (a) software, derived from a detailed analysis of what differentiates information contained in historical sources from such present in current material, (b) databases which are as easily available as printed books and (c) knowledge bases which allow software and data bases to draw upon the information contained in historical reference works. A loose network of European research projects, dedicated to the realization of such a setup, is described.Manfred Thaller has a Ph.D. (1975) in Modern and Medieval History and held a post-doctoral fellowship in sociology at the Institute for Advanced Studies (Vienna). Since 1978 he has been at the Max-Planck-Institute for History, where he is a research fellow for Historical Information Science.     

Effect of environmental humidity and coating on aroma transfer through treated papers

Coupled transfers of aroma compounds and water vapour were investigated by varying the relative humidity gradient of storage (50% or 90%) of two paper packaging at 25 °C. These papers differed in their coating surface: both were identically impregnated then supercalendered, and only one was twice coated on both sides with a synthetic barrier substance. Permeability and solubility coefficients were determined. The coating treatment was more effective to decrease the permeabilities of water vapour and ethyl ester than the effect of RH. On the contrary, the RH modified the water content of the treated papers and affected more strongly their permeability and solubility to cis-3-hexenol and benzaldehyde. The cis-3-hexenol transfer through the non-coated paper and the benzaldehyde transfer through the coated paper decreased due to a probable competition with sorbed water although it increases for the cis-3-hexenol of a plasticisation phenomenon.     

烘焙提香时间对工夫红茶品质的影响

为提高工夫红茶的甜香度,本文以山东夏季鲜叶加工的工夫红茶初制品为原料,在70℃的烘温下进行不同时间的烘焙提香处理,并通过感官审评、生化成分和香气分析方法研究其对工夫红茶品质的影响,结果表明:烘焙时间的长短不仅影响工夫红茶的香型和高低,而且也影响汤色和滋味,在本试验条件下,烘焙时间以1.0~1.5h的感官品质较好;此外,烘焙时间对工夫红茶各生化成分的影响不同,但对品质有利的生化成分以烘焙1.0h的茶样含量最高;随着烘焙时间的延长,工夫红茶香气的种类、含量以及组分比值发生显著变化,从而可以解释不同烘焙时间工夫红茶香型存在差异的原因。     

The effects of extrusion conditions and the addition of volatile compounds and flavour enhancers to corn grits on the retention of the volatile compounds and texture of the extrudates

Corn grits that were supplemented with isovaleraldehyde, ethyl butyrate, butyric acid and flavour enhancers were extruded under different processing conditions. Volatile compounds retained in the extrudates were isolated by dynamic headspace and analysed using gas chromatography–mass spectrometry. The expansion ratio, density and cut force to break down the extrudates were evaluated and aroma intensity was assessed using a multisample difference test. Butyric acid showed the greatest retention (96.4%), regardless of the extrusion conditions. All compounds were better retained when samples were extruded at 20% feed moisture and 90 °C processing temperature (2.9–81.0%), conditions that also resulted in greater aromatic intensity (moderate to moderate‐strong intensity). The addition of volatile compounds reduced the expansion ratio and cut force, whereas the addition of flavour enhancers increased the expansion ratio but reduced ethyl butyrate and butyric acid retention.     

A global study using PANAS (PA and NA) scales to measure consumer emotions associated with aromas of phytonutrient supplements

Phytonutrient supplements are consumed to increase dietary consumption of nutrients. This research addresses the opportunities and the challenges for this product category to better understand consumer choice and consumption in combination with hedonic (liking), sensory, psychographic and emotion measures. One objective of this exploratory study is to use Positive Affect Negative Affect Schedule (PANAS) to measure consumer (n = 303) emotions associated with 9 different aromas of phytonutrient supplements.A key issue facing emotion research in product development is the proper balance of positive and negative emotions. Traditional clinical emotion questionnaires emphasize negative emotions. Newer emotion questionnaires for commercial use emphasize positive emotions, both in the food and perfume categories. What is the proper balance of positive and negative emotions for a broad range of consumer products? Desmet and Schifferstein (2008) noted the “hedonic asymmetry”, or positive bias, which exists for many product emotions. In contrast to the clinical questionnaires which emphasize negative emotions, and the newer commercial questionnaires which emphasize positive emotions, the Positive Affect and Negative Affect Schedule (PANAS) has equal numbers of positive and negative emotions in both the regular and shorter forms.This study supports that the PANAS scales are valid in measurement of consumer emotions evoked by the aromas of phytonutrient supplements. The PANAS scale with 20-items and the short PANAS scale with 10-items are largely consistent in the results. Different phytonutrient product aromas were discriminated on their PANAS emotional profiles, and on both positive (PA) and negative (NA) dimensions. Results suggest that the hedonic, sensory, and emotional attributes represent different dimensions in consumer choice and consumption behaviors.     

Impact of Debaryomyces hansenii strains inoculation on the quality of slow dry-cured fermented sausages

Debaryomyces hansenii strains, M4 and P2, isolated from natural fermented sausages were inoculated in slow fermented sausages to study their effect on processing parameters, microbial population, volatile compound and sensory characteristics. The inoculation of D. hansenii strains, M4 and P2, did not affect the ripening process as no differences in pH and Aw were detected. The dominance of the inoculated yeast strains along the process was followed by RAPDs of M13 minisatellite. The inoculated yeasts, P2 and M4, were recovered at the end of the ripening process although P2 appeared in higher counts than M4. The sausages inoculated with P2 resulted in a decrease in lipid oxidation values (TBARS) and a reduction of lipid-oxidation derived aldehydes in addition to a highest acid compound abundance. M4 inoculated sausages resulted in highest sulphur containing compound abundance. However, no differences in consumer acceptance were detected. Moreover, both yeast strains were responsible for the generation of ethyl methyl-branched ester compounds in the dry-cured sausages.