ESI-mass spectrometry analysis of unsubstituted and disubstituted β-cyclodextrins: fragmentation mode and identification of the AB, AC, AD regioisomers

AbstractThe study of unsubstituted and disubstituted β-cyclodextrins (β-CDs) by ESI-mass spectrometry is reported, applying a cone-induced fragmentation in the presence of a twofold excess of sodium chloride, in order to gain information about the fragmentation of the different regioisomers. On the basis of the fragmentation pattern observed for the unsusbstituted β-CD, a statistical model shows that the fragments generated by every regioisomer of a disubstituted CD (AB, AC, and AD) are expected to differ in their relative intensity and, therefore, they can be used for correctly identifying the three different regioisomers. The model was tested on the three regioisomeric (AB, AC, and AD) diamino-β-CDs and ditosyl-β-CD and on the AC and AD regioisomers of dimesitylenesulphonyl-β-CD, allowing in every case through statistical analysis of the fragmentation pattern the correct assignment of every regioisomer on the basis of an ESI mass spectrum (single quadrupole analyzer, high cone voltage) of the pure compounds. The absolute intensities of the fragmentation peaks were voltage-dependent but their ratios was voltage-independent, indicating that no mass bias in peak ratios is introduced by the analyzer. Given the fast time of analysis and its general applicability, independently from the substituents, we propose our method as an easy way to identify the regioisomers of disubstituted β-CDs

Degree of Hydrolysis Affects the Techno-Functional Properties of Lesser Mealworm Protein Hydrolysates

Protein hydrolysates from lesser mealworm (Alphitobius diaperinus, LM) were obtained by enzymatic hydrolysis with protease from Bacillus licheniformis. A preliminary test performed for five hours of hydrolysis generated an insect protein hydrolysate with 15% of degree of hydrolysis (DH), optimum solubility property and oil holding capacity, but emulsifying and foaming ability were completely impaired. In order to investigate the potential implication of DH on techno-functional properties, a set of protein hydrolysates with a different DH was obtained by sub-sampling at different time points during three hours of enzymatic hydrolysis process. An increase in DH% had positive effects on the solubility property and oil holding ability, while a reduced emulsifying ability was observed up to five hours of hydrolysis. These results demonstrated that the enzymatic hydrolysis, if performed under controlled conditions and not for a long period, represents a valid method to extract high quality protein from insects with tailored techno-functionality, in order to produce tailored ingredients for feed and food purpose

Effect of Maillard induced glycation on protein hydrolysis by lysine/arginine and non-lysine/arginine specific proteases

Enzymatic protein hydrolysis is sensitive to modifications of protein structure, e.g. Maillard reaction. In early stages of the reaction glycation takes place, modifying the protein primary structure. In later stages protein aggregation occurs. The specific effect of glycation on protein hydrolysis was studied using α-lactalbumin glycated with D-glucose at 50 °C (0–10 h). This resulted in proteins with different degrees of glycation (DG = 0–63%) without changes in secondary, tertiary and quaternary structure. These glycated proteins were hydrolyzed by lysine/arginine specific proteases (bovine and porcine trypsin) or by non-lysine/arginine specific proteases (Bacillus licheniformis protease (BLP), α-chymotrypsin and subtilisin A). For bovine and porcine trypsin, the maximal degree of hydrolysis decreased linearly with 65% from untreated to maximal glycated protein (DG = 63%). This means trypsin cannot hydrolyze glycated cleavage sites. BLP and subtilisin A hydrolyses were independent of glycation, while α-chymotrypsin cannot hydrolyze cleavage sites with glycated binding sites. This means for non-lysine/arginine specific proteases, the effect of glycation depends on the enzyme sensitivity towards modifications on binding sites. Since not all cleavage sites are efficiently used by the enzymes, the extent of the effects depends on the enzyme selectivity towards cleavage sites (for trypsin) or cleavage sites near glycation sites (for α-chymotrypsin). Combining the results of all proteases, an equation was derived describing the effect of modification of protein primary structure on the extent of hydrolysis based on the enzyme specificity, selectivity and binding site sensitivity

Influence of the killing method of the black soldier fly on its lipid composition

Black soldier fly (BSF, Hermetia illucens) represents a valuable source of biomolecules and it also constitutes an economic way to valorise residual biomasses. BSF prepupae contain high amounts of lipids (37% DM basis). The present investigation aimed at studying the composition of BSF lipids and the effect of killing/storage on their quality. The main fatty acid was lauric acid, sterols were represented primarily by beta-sitosterol and campesterol. Global fatty acid and sterol profiles, determined by GC–MS, were only slightly affected by the killing procedure, while lipid classes distribution, determined by 1H NMR, strongly changed. Prepupae killed by freezing showed a drastic reduction of acylglycerols during storage and a relevant release of free fatty acids, likely due to activation of lipases. On the contrary, prepupae killed by blanching have a stable lipid fraction constituted mainly by triacylglycerols. Therefore, killing procedure strongly influences BSF oil composition and the potential applications

Standardized wooden system for timber internal tension simulation

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Effects of pulmonary rehabilitation on exercise capacity in patients with COPD: A number needed to treat study

BACKGROUND: Pulmonary rehabilitation (PR) is recognized as an evidence-based treatment in improving dyspnea and quality of life in patients with COPD. We evaluated the number needed to treat (NNT) to achieve an increase in physical capacity, as defined by a significant improvement in the six-minute walk test (6MWT) in patients with COPD undergoing PR. METHODS: The study enrolled 284 patients aged 41 to 86 years (mean age 69.4 years) divided into two groups: a study group (222 patients) undergoing a PR program, and a control group (62 patients) treated only with drugs. The study group included patients with COPD divided in four subgroups according to GOLD stages. RESULTS: In the study group, 142 out of 222 patients (64%) had an increase of at least 54 m in the 6MWT following PR versus 8 out of 62 patients (13%) in the control group after the same time interval. The NNT in the overall study group was 2; the same NNT was obtained in GOLD stages 2, 3, and 4, but was 8 in stage 1. CONCLUSIONS: PR is highly effective in improving the exercise capacity of patients with COPD, as demonstrated by a valuable NNT, with better results in patients with a more severe disease

Protein Quality and Protein Digestibility of Vegetable Creams Reformulated with Microalgae Inclusion

Microalgae are considered a valuable source of proteins that are used to enhance the nutritional value of foods. In this study, a standard vegetable cream recipe was reformulated through the addition of single-cell ingredients from Arthrospira platensis (spirulina), Chlorella vulgaris, Tetraselmis chui, or Nannochloropsis oceanica at two levels of addition (1.5% and 3.0%). The impact of microalgae species and an addition level on the amino acid profile and protein in vitro digestibility of the vegetable creams was investigated. The addition of microalgae to vegetable creams improved the protein content and the amino acid nutritional profile of vegetable creams, whereas no significant differences were observed in protein digestibility, regardless of the species and level of addition, indicating a similar degree of protein digestibility in microalgae species despite differences in their protein content and amino acid profile. This study indicates that the incorporation of microalgae is a feasible strategy to increase the protein content and nutritional quality of foods.This research was funded by the ProFuture project (2019–2023 “Microalgae protein-rich ingredients for the food and feed of the future”). The ProFuture project received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 862980. CERCA Programme (Generalitat de Catalunya) also supported this research.info:eu-repo/semantics/publishedVersio

Antimicrobial Biomasses from Lactic Acid Fermentation of Black Soldier Fly Prepupae and Related By-Products

Worldwide, thousands of insect species are consumed as food or are used as feed ingredients. Hermetia illucens, ‘black soldier fly’, is one of them, and a large amount of puparia and dead adults flies are accumulated during rearing. These materials represent important wastes but no studies are still present in the literature regarding their functional properties and potential reuse. Lactic acid bacteria (LAB) are a heterogeneous group of bacteria contributing to various industrial applications, ranging from food fermentation, chemicals production to pharmaceuticals manufacturing. A LAB feature of industrial interest is their ability to produce antimicrobial metabolites. Considering the scientific and commercial interest in discovering novel antimicrobials, this work will be direct towards fermentation of insect-derived biomasses: puparia and adults insect at the end of life cycle. To the best of our knowledge, the in vitro antimicrobial activity of fermented insects is tested for the first time. This study aimed also to evaluate differences in the composition between fermented and unfermented insects, and to study whether the fermentation and the type of LAB used played a crucial role in modifying the composition of the substrate. Results firstly highlighted fermentability of this species of insects, showed that fermented black soldier flies puparium possess a high antimicrobial activity against tested pathogens. Moreover, result of chemical composition showed that fermented biomass had a higher percentage of fat and a more complex fatty acids profile