Efficient generation of neural stem cell-like cells from adult human bone marrow stromal cells

Clonogenic neural stem cells (NSCs) are self-renewing cells that maintain the capacity to differentiate into brain-specific cell types, and may also replace or repair diseased brain tissue. NSCs can be directly isolated from fetal or adult nervous tissue, or derived from embryonic stem cells. Here, we describe the efficient conversion of human adult bone marrow stromal cells (hMSC) into a neural stem cell-like population (hmNSC, for human marrow-derived NSC-like cells). These cells grow in neurosphere-like structures, express high levels of early neuroectodermal markers, such as the proneural genes NeuroD1, Neurog2, MSl1 as well as otx1 and nestin, but lose the characteristics of mesodermal stromal cells. In the presence of selected growth factors, hmNSCs can be differentiated into the three main neural phenotypes: astroglia, oligodendroglia and neurons. Clonal analysis demonstrates that individual hmNSCs are multipotent and retain the capacity to generate both glia and neurons. Our cell culture system provides a powerful tool for investigating the molecular mechanisms of neural differentiation in adult human NSCs. hmNSCs may therefore ultimately help to treat acute and chronic neurodegenerative diseases

FT-ICR-MS reveals the molecular imprints of the brewing process

The study of fermentation and brewing has a long history of pioneering discoveries that continue to influence modern industrial food production. Since then, numerous research endeavors have yielded conventional criteria that guide contemporary brewing practices. However, the intricate open challenges faced today necessitate a more exhaustive understanding of the process at the molecular scale. We have developed an ultra-high-resolution mass spectrometric analysis (FT-ICR-MS) of the brewing process that can rapidly and comprehensively resolve thousands of molecules. This approach allows us to track molecular fluctuation during brewing at the level of chemical compositions. Employing biological triplicates, our investigation of two brewing lines that are otherwise identical except for the malt used revealed over 8,000 molecular descriptors of the brewing process. Metabolite imprints of both the similarities and differences arising from deviating malting temperatures were visualized. Additionally, we translated traditional brewing attributes such as the EBC-value, free amino nitrogen, pH-value, and concentration curves of specific molecules, into highly correlative molecular patterns consisting of hundreds of metabolites. These in-depth molecular imprints provide a better understanding of the molecular circumstances leading to various changes throughout the brewing process. Such chemical maps go beyond the observation of traditional brewing attributes and are of great significance in the investigation strategies of current open challenges in brewing research. The molecular base of knowledge, along with advancements in technological and data integration schemes, can facilitate the efficient monitoring of brewing and other productions processes

Screening of Mycotoxigenic Fungi in Barley and Barley Malt (Hordeum vulgare L.) Using Real-Time PCR—A Comparison between Molecular Diagnostic and Culture Technique

Filamentous fungi have a crucial impact on the food safety and technological quality of malting barley. Commonly used techniques for the detection of seed-borne fungi are based on cultivation and identification by morphological criteria. In contrast, this study established a quantitative real-time polymerase chain reaction (PCR) assay based on SYBR green technology for the detection and quantification of black fungal species (Alternaria spp., Epicoccum nigrum, Cladosporium cladosporioides, Penicillium verrucosum and Aspergillus niger) on brewing barley and compares it with the traditional cultivation technique and visual assessment. To screen the fungal spectrum over different barley varieties and harvest years, naturally infected samples of malting barley and corresponding malts (Hordeum vulgare L.) were analyzed over four consecutive years (2018–2021), grown under different climatic conditions in Germany. Alternaria and Cladosporium spp. DNA were present in all examined barley samples, even without visible contamination. In contrast, detection via culture-based methods does not reliably cover all species. Molecular analysis showed that there was less fungal biomass after malting, by 58.57% in the case of A. alternata, by 28.27% for Cladosporium spp. and by 12.79% for Epicoccum nigrum. Correlation analysis showed no causal relationship between fungal DNA and the number of black kernels. The qPCR provides a highly sensitive and time-saving screening method for detecting latent fungal infections in brewing grains to identify batches that are potentially highly contaminated with toxigenic fungi

Gelatinization or Pasting? The Impact of Different Temperature Levels on the Saccharification Efficiency of Barley Malt Starch

Efficient enzymatic hydrolysis of cereal starches requires a proper hydrothermal pre-treatment. For malted barley, however, the exact initial temperature is presently unknown. Therefore, samples were micro-mashed according to accurately determined gelatinization and pasting temperatures. The impact on starch morphology, mash viscometry and sugar yields was recorded in the presence and absence of an amylase inhibitor to differentiate between morphological and enzymatic effects. Mashing at gelatinization onset temperatures (54.5–57.1 °C) led to negligible morphological and viscometric changes, whereas mashing at pasting onset temperatures (57.5–59.8 °C) induced significant starch granule swelling and degradation resulting in increased sugar yields (61.7% of upper reference limit). Complete hydrolysis of A-type and partial hydrolysis of B-type granules was achieved within only 10 min of mashing at higher temperatures (61.4–64.5 °C), resulting in a sugar yield of 97.5% as compared to the reference laboratory method mashing procedure (65 °C for 60 min). The results indicate that the beginning of starch pasting was correctly identified and point out the potential of an adapted process temperature control

The Influence of Proteolytic Malt Modification on the Aging Potential of Final Wort

The dynamic changes in beer flavor are determined by its aging potential, which comprises of present free and bound-state aldehydes and their precursors. Rising flavor-active aging compounds cause sensory deterioration (flavor instability). These compounds are mainly formed upstream in the brewing process through the Maillard reaction, the Strecker degradation, or lipid oxidation. Wort boiling is an especially critical production step for important reactions due to its high temperature and favorable pH value. Amino acid concentration, as an important aging-relevant precursor, is variable at the beginning of wort boiling, mainly caused by the malt modification level, and can further influence the aging potential aging formation during wort boiling. This study investigated the effect of the proteolytic malt modification level on the formation of precursors (amino acids and dicarbonyls) and free and bound-state aldehydes during wort boiling. Six worts (malt of two malting barley varieties at three proteolytic malt modification levels) were produced. Regarding precursors, especially Strecker, relevant amino acids and dicarbonyls increased significantly with an enhanced malt modification level. Concentrations of free and bound aldehydes were highest at the beginning of boiling and decreased toward the end. A dependency of malt modification level and the degree of free and bound aldehydes was observed for 2-methylpropanal, 2-methylbutanal, and 3-methylbutanal. Generally, a higher proteolytic malt modification level tended to increase free and bound aldehyde content at the end of wort boiling. Conclusively, the aging potential formation during boiling was increased by an intensified malt modification level

Multi-Response Optimization of the Malting Process of an Italian Landrace of Rye (<i>Secale cereale</i> L.) Using Response Surface Methodology and Desirability Function Coupled with Genetic Algorithm

Rye is used in some applications in the food and beverage industry and for the preparation of functional foods. It is an interesting raw material in malting and brewing due to its characteristic contribution to the beer’s color, turbidity, foam and aroma. The aim of this work was to optimize the micro-malting process of a rye landrace. The response surface methodology (RSM) was applied to study the influence of three malting parameters (germination time, germination temperature and degree of steeping) on the quality traits of malted rye. Long germination times at high temperatures resulted in an increase in the extract and Kolbach index. The model for the apparent attenuation limit showed a particular pattern, whereby time and temperature inversely influenced the response. The lowest viscosities were determined in the worts produced from highly modified malts. Optimization of the variables under study was achieved by means of a desirability function and a genetic algorithm. The two methodologies provided similar results. The best combination of parameters to optimize the malting process on the rye landrace under study was achieved at 6 days, 12 °C and 44 g/100 g

A Comprehensive Evaluation of Flavor Instability of Beer (Part 2): The Influence of De Novo Formation of Aging Aldehydes

Flavor instability of beer is affected by the rise of aroma-active aldehydes during aging. Aldehydes can be either released from bound-state forms or formed de novo. This second part of our study focused on the de novo formation of aldehydes during the Maillard reaction, Strecker degradation, and oxidation reactions. Key precursor compounds for de novo pathways are free amino acids. This study varied the potential for reactions by varying free amino acid content in fresh beer using different proteolytic malt modification levels (569–731 mg/100 g d. m. of soluble nitrogen) of the used malt in brewing trials. Overall, six pale lager beers were produced from three malts (different malt modification levels), each was made from two different barley varieties and was naturally and forcibly aged. It was found that higher malt modification levels in fresh beer and during beer aging increased amino acid and dicarbonyl concentrations as aging precursors and Strecker aldehyde contents as aging indicators. Dicarbonyls were degraded during aging. Advanced glycation end products as possible degradation products showed no consistent formation during aging. Therefore, Strecker reactions were favored during beer aging. No alternative oxidative formation of Strecker aldehydes from their corresponding alcohols could be confirmed. Along with the preceding part one of our investigation, the results of this study showed that de novo formation and release occur simultaneously. After 4 months of natural aging, aldehyde rise is mainly accounted for by de novo formation

Data_Sheet_1_FT-ICR-MS reveals the molecular imprints of the brewing process.PDF

The study of fermentation and brewing has a long history of pioneering discoveries that continue to influence modern industrial food production. Since then, numerous research endeavors have yielded conventional criteria that guide contemporary brewing practices. However, the intricate open challenges faced today necessitate a more exhaustive understanding of the process at the molecular scale. We have developed an ultra-high-resolution mass spectrometric analysis (FT-ICR-MS) of the brewing process that can rapidly and comprehensively resolve thousands of molecules. This approach allows us to track molecular fluctuation during brewing at the level of chemical compositions. Employing biological triplicates, our investigation of two brewing lines that are otherwise identical except for the malt used revealed over 8,000 molecular descriptors of the brewing process. Metabolite imprints of both the similarities and differences arising from deviating malting temperatures were visualized. Additionally, we translated traditional brewing attributes such as the EBC-value, free amino nitrogen, pH-value, and concentration curves of specific molecules, into highly correlative molecular patterns consisting of hundreds of metabolites. These in-depth molecular imprints provide a better understanding of the molecular circumstances leading to various changes throughout the brewing process. Such chemical maps go beyond the observation of traditional brewing attributes and are of great significance in the investigation strategies of current open challenges in brewing research. The molecular base of knowledge, along with advancements in technological and data integration schemes, can facilitate the efficient monitoring of brewing and other productions processes.</p

Sensory and Olfactometry Chemometrics as Valuable Tools for Assessing Hops’ Aroma Impact on Dry-Hopped Beers: A Study with Wild Portuguese Genotypes

Sensory, olfactometry (using the sums of odour intensities for each class of compounds) and chemometric analyses were used to evaluate Portuguese wild hops’ sensory characteristics and the aroma that those hops impart to dry-hopped beer. CATA analysis and agglomerative hierarchical clustering was applied for the sensory characterization of 15 wild hops of Portuguese genotypes, clustering them in two groups: one more sulphurous, floral, and fruity, and another more earthy, resinous, floral, and non-citrus fruits. Two hops representative of each group were selected for the production of four dry-hopped beers using the same base beer style (Munich Helles). Beers were analysed by quantitative descriptive analyses and quantification of hop-derived key volatile compounds. Multivariate statistical treatment of the data was performed. Results indicate significant differences (p &lt; 0.05) in fruity, resinous, earthy, floral, and sulphurous attributes of hops, but the dry-hopped beers only have a significant increase (p &lt; 0.05) in fruity and spicy notes when compared with non-dry-hopped Munich-style Helles beer. Hop olfactometry explained the sensory perception that the 11 hops not used for brewing (employed as supplementary observations) are placed into the space of the odour-active compounds profile of the four hops selected for brewing. These 11 hop samples have more spiciness than fruitiness potential

Fate of <i>Fusarium</i> Toxins during the Malting Process

Little is known about the fate of Fusarium mycotoxins during the barley malting process. To determine the fungal DNA and mycotoxin concentrations during malting, we used barley grain harvested from field plots that we had inoculated with Fusarium species that produce type A or type B trichothecenes or enniatins. Using a recently developed multimycotoxin liquid chromatography–tandem mass stable isotope dilution method, we identified Fusarium-species-specific behaviors of mycotoxins in grain and malt extracts and compared toxin concentrations to amounts of fungal DNA in the same samples. In particular, the type B trichothecenes and Fusarium culmorum DNA contents were increased dramatically up to 5400% after kilning. By contrast, the concentrations of type A trichothecenes and Fusarium sporotrichioides DNA decreased during the malting process. These data suggest that specific Fusarium species that contaminate the raw grain material might have different impacts on malt quality