Impact of ethanol content on the scavenging activities of oak wood C‐glycosidic ellagitannins. Application to the evaluation of the nutritional status of spirits

As a consequence of their structure, ellagitannins have a high level of activity in terms of their oxidative properties. In particular, their scavenging activity is often presented as a noteworthy parameter, since polyphenols have a beneficial impact on human health. A large variety of spirits, brandies and whiskeys are aged in barrels or receive a wood extract rich in polyphenols. Although spirits (alcoholic beverages) represent a very dynamic market with significant growth, very few scientific papers have been published on this topic, or have justified specific research in this field. Firstly, the scavenging properties of ellagitannins when measured in a solution are presented. An oak wood extract was fractionated by gel chromatography to obtain different fractions with different polymeric levels. It is particularly clear that the oligomer forms are the most efficient by comparison with low‐weight and high‐weight fractions. A mixture of ellagitannins has a higher scavenging intensity than the pure molecule; this illustrates the synergic effect of polyphenols. The scavenging activity of different commercial spirits depends on the presence and quantities of ellagitannins. As an example, the percentage of scavenged superoxide anion radicals represents 70 and 18%, respectively, for spirits aged in a barrel (12 years) and white spirits. In a second step, details of the influence of ethanol on the antioxidant properties of ellagitannins are provided. In particular, alcohol increases the scavenging activity of ellagitannins from 0 to 25% vol., and higher concentrations present a certain degree of pro‐oxidant activity. The chemical study of ellagitannin scavenging properties in solution is confirmed by means of a specific in vitro investigation on the DNA extracted from cells, and directly on the cells. Copyright © 2013 The Institute of Brewing & Distilling     

Surface Binding of TOTAPOL Assists Structural Investigations of Amyloid Fibrils by Dynamic Nuclear Polarization NMR Spectroscopy

Dynamic nuclear polarization (DNP) NMR can enhance sensitivity but often comes at the price of a substantial loss of resolution. Two major factors affect spectral quality: low‐temperature heterogeneous line broadening and paramagnetic relaxation enhancement (PRE) effects. Investigations by NMR spectroscopy, isothermal titration calorimetry (ITC), and EPR revealed a new substantial affinity of TOTAPOL to amyloid surfaces, very similar to that shown by the fluorescent dye thioflavin‐T (ThT). As a consequence, DNP spectra with remarkably good resolution and still reasonable enhancement could be obtained at very low TOTAPOL concentrations, typically 400 times lower than commonly employed. These spectra yielded several long‐range constraints that were difficult to obtain without DNP. Our findings open up new strategies for structural studies with DNP NMR spectroscopy on amyloids that can bind the biradical with affinity similar to that shown towards ThT.     

Antibiotic susceptibility of Bifidobacterium thermophilum and Bifidobacterium pseudolongum isolates from animal sources

The widespread use of antimicrobial substances has led to resistant populations of microorganisms in several ecosystems. In animal husbandry, the application of antibiotics has contributed to resistance development in pathogenic and commensal bacteria. These strains or their resistance genes can be spread along several ecological routes, including the food chain. Antibiotic resistance is important in terms of the safety of industrial strains, such as probiotics for food and feed. Bifidobacterium thermophilum and Bifidobacterium pseudolongum are known to comprise the major part of the bifidobacterial microbiota in the gut and feces of cattle and pigs. In this study, the antimicrobial susceptibility in bifidobacterial isolates of these species was investigated. Isolates from the beef and pork production chain were identified and typed to strain level, and the antimicrobial susceptibility level was tested to a set of antibiotics. Isolates with low susceptibility levels were screened by PCR for already described resistance genes. Strains atypically resistant to clindamycin, erythromycin, and tetracycline were determined. The resistance genes tet(O), tet(W), and erm(X) were detected in the bifidobacterial species that were examined.     

Differentiated Patterns of Legume Sensitisation in Peanut-Allergic Patients

Patients with peanut allergy are often serologically sensitised to other legumes. If primary peanut allergy was contracted, secondary allergies to other legume plants may develop. The cross-sensitisation implies an enhanced risk of food-related anaphylaxis for the patients even if its clinical relevance is under discussion. In the present study, 32 patients with anti-peanut immunoglobulin E-levels (IgE) of at least class 1 according to the ImmunoCap™ classification system were included. The patients had been referred to the Norwegian National Register and Reporting System for Severe Allergic Reactions to Food with histories of anaphylaxis by the consumption of legume-containing foods. The patient sera were analysed for IgE that reacted to peanut (Arachis hypogea), soybean (Glycine max), pea (Pisum sativum), lupin (Lupinus albus) and fenugreek (Trigonella foenum graecum). Immunoblot experiments were performed using extracts of in total eight different legume plants. IgE measurements and blot analyses revealed differentiated patterns of legume sensitisation. One third of the patients had anti-peanut IgE class 4 to 6. In contrast, just one patient had specific IgE higher than class 4 to respectively pea, soybean and fenugreek. However, ten patients had elevated anti-fenugreek IgE (≥class 3), significantly more than for the other legumes. The second highest levels were found for soybean, followed by pea and lupin. The immunoblots showed particularly strong IgE binding to proteins at 12, 22, 30, 36 and 50 kDa for all legumes, indicating that the major peanut allergens Ara h 1, Ara h 2 and Ara h 3 are strong candidates for causing cross-sensitivity.     

Pre-Omicron Vaccine Breakthrough Infection Induces Superior Cross-Neutralization against SARS-CoV-2 Omicron BA.1 Compared to Infection Alone

SARS-CoV-2 variants raise concern because of their high transmissibility and their ability to evade neutralizing antibodies elicited by prior infection or by vaccination. Here, we compared the neutralizing abilities of sera from 70 unvaccinated COVID-19 patients infected before the emergence of variants of concern (VOCs) and of 16 vaccine breakthrough infection (BTI) cases infected with Gamma or Delta against the ancestral B.1 strain, the Gamma, Delta and Omicron BA.1 VOCs using live virus. We further determined antibody levels against the Nucleocapsid (N) and full Spike proteins, the receptor-binding domain (RBD) and the N-terminal domain (NTD) of the Spike protein. Convalescent sera featured considerable variability in the neutralization of B.1 and in the cross-neutralization of different strains. Their neutralizing capacity moderately correlated with antibody levels against the Spike protein and the RBD. All but one convalescent serum failed to neutralize Omicron BA.1. Overall, convalescent sera from patients with moderate disease had higher antibody levels and displayed a higher neutralizing ability against all strains than patients with mild or severe forms of the disease. The sera from BTI cases fell into one of two categories: half the sera had a high neutralizing activity against the ancestral B.1 strain as well as against the infecting strain, while the other half had no or a very low neutralizing activity against all strains. Although antibody levels against the spike protein and the RBD were lower in BTI sera than in unvaccinated convalescent sera, most neutralizing sera also retained partial neutralizing activity against Omicron BA.1, suggestive of a better cross-neutralization and higher affinity of vaccine-elicited antibodies over virus-induced antibodies. Accordingly, the IC50: antibody level ratios were comparable for BTI and convalescent sera, but remained lower in the neutralizing convalescent sera from patients with moderate disease than in BTI sera. The neutralizing activity of BTI sera was strongly correlated with antibodies against the Spike protein and the RBD. Together, these findings highlight qualitative differences in antibody responses elicited by infection in vaccinated and unvaccinated individuals. They further indicate that breakthrough infection with a pre-Omicron variant boosts immunity and induces cross-neutralizing antibodies against different strains, including Omicron BA.1.     

An Experimental Comparison of the Maintainability of Object-Oriented and Structured Design Documents

Several important questions still need to be answered regarding the maintainability of object-oriented design documents. This paper focuses on the following issues: are object-oriented design documents easier to understand and modify than structured design documents? Do they need to comply with quality guidelines such as the ones provided by Coad and Yourdon? What is the impact of such quality standards on the understandability and modifiability of design documents? Answers can be based on informed opinion or empirical evidence. Since software technology investments are substantial and contradictory opinions exist regarding design strategies, performing empirical studies on these topics is a relevant research activity. This paper presents a controlled experiment performed with computer science students as subjects. Results strongly suggest that quality guidelines based on Coad and Yourdon principles have a beneficial effect on the maintainability of object-oriented design documents. However, there is no strong evidence regarding the alleged higher maintainability of object-oriented design documents over structured design documents. Furthermore, results suggest that object-oriented design documents are more sensitive to poor design practices, in part because their cognitive complexity becomes increasingly unmanageable. However, because our ability to generalise these results is limited, they should be considered as preliminary, i.e., it is very likely that they can only be generalised to programmers with little object-oriented training and programming experience. Such programmers can, however, be commonly found on maintenance projects. As well as additional research, external replications of this study are required to confirm the results and achieve confidence in these findings.     

Getting a Grip on the Undrugged: Targeting β-Catenin with Fragment-Based Methods

Aberrant WNT pathway activation, leading to nuclear accumulation of β-catenin, is a key oncogenic driver event. Mutations in the tumor suppressor gene APC lead to impaired proteasomal degradation of β-catenin and subsequent nuclear translocation. Restoring cellular degradation of β-catenin represents a potential therapeutic strategy. Here, we report the fragment-based discovery of a small molecule binder to β-catenin, including the structural elucidation of the binding mode by X-ray crystallography. The difficulty in drugging β-catenin was confirmed as the primary screening campaigns identified only few and very weak hits. Iterative virtual and NMR screening techniques were required to discover a compound with sufficient potency to be able to obtain an X-ray co-crystal structure. The binding site is located between armadillo repeats two and three, adjacent to the BCL9 and TCF4 binding sites. Genetic studies show that it is unlikely to be useful for the development of protein–protein interaction inhibitors but structural information and established assays provide a solid basis for a prospective optimization towards β-catenin proteolysis targeting chimeras (PROTACs) as alternative modality.     

Modulation of Transaminase Activity by Encapsulation in Temperature-Sensitive Poly(N-acryloyl glycinamide) Hydrogels

Smart hydrogels hold much potential for biocatalysis, not only for the immobilization of enzymes, but also for the control of enzyme activity. We investigated upper critical solution temperature-type poly N-acryloyl glycinamide (pNAGA) hydrogels as a smart matrix for the amine transaminase from Bacillus megaterium (BmTA). Physical entrapment of BmTA in pNAGA hydrogels results in high immobilization efficiency (>89 %) and high activity (97 %). The temperature-sensitiveness of pNAGA is preserved upon immobilization of BmTA and shows a gradual deswelling upon temperature reduction. While enzyme activity is mainly controlled by temperature, deactivation tended to be higher for immobilized BmTA (≈62–68 %) than for free BmTA (≈44 %), suggesting a deactivating effect due to deswelling of the pNAGA gel. Although the deactivation in response to hydrogel deswelling is not yet suitable for controlling enzyme activity sufficiently, it is nevertheless a good starting point for further optimization.     

Lignin and lignin derivatives as components in biobased hot melt adhesives

Hot melt adhesives (HMAs) are formulated for the first time with different lignins as major components, and the developed HMA formulations were tested for gluing paperboard. The best formulations showed equal or even better bond strength compared to a commercial HMA reference. A maximum bond strength of 16.1 N was achieved with a formulation of oxidized cellulose acetate, organosolv lignin, and triethyl citrate, whereas the bond strength of the commercial HMA reference was 10.5 N. The performance was adjusted via the selection of lignin, the formulation, and chemical modification. Lignin modification was not necessary but provided further possibilities for adjusting the properties for different products (reversible vs irreversible adhesive seams) and also for producing plasticizer-free formulations. Modification with tall oil fatty acids enabled the formulation of fully biobased HMAs without any external plasticizer and provided a bond strength as high as high as 8.9 N. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47983.