Synthesis,Characterization and Application of Sucrose Carboxylic Acid Polyesters — A Prototype of Non-Caloric or Surface Active Compounds

Sucrose carboxylic acid polyesters are in dependence of the degree of acylation and fatty acid composition more or less undigestible or emulsifying fat substitutes and functional additives for foods. In this connection the relationships between their molecular parameters and functional properties are discussed. New procedures of the solvent-free synthesis are presented. Furthermore, open biochemical and toxicological problems of pinocytosis and accumulation of intact polyesters in organs and tissue being still under investigations are considered.     

SDS electrophoresis of legume seed proteins in horizontal ultrathin-layer pore gradient gels

Summary Horizontal SDS electrophoresis of 18 legume seed protein extracts was performed in ultrathin-layer polyacrylamide gels on foil supports. Separation results of the SD S pore gradient electrophoresis (T=4–22.5%) are compared to those of SDS electrophoresis in a constant pore size gel (T=10%). Resolution as well as the sensitivity (0.1 g protein per band) of the ultrathin-layer SDS pore-gradient electrophoresis were extremely high. Because of the very low gel thickness, separation, staining and drying were completed in substantially shorter times than achieved with conventional thick gels. An easy technique for casting ultrathin-layer (360 gm) concave gradient gels for 10 cm separation distance and a width of 25 cm is described. The even distribution of the concave exponential pore-gradient over the whole gel width is demonstrated. Molecular weights of the legume proteins are detected from 5,000 to 110,000 daltons. The protein patterns are genus- and species-specific.

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Horizontale Ultradünnschicht SDS-Gradientengel-Elektrophorese von Leguminosensamenproteinen

Zusammenfassung Die ausgezeichnete Trennschärfe der horizontalen Ultradünnschicht-SDS-Gradienten-gel Elektrophorese wird am Beispiel von Samenproteinen 18 verschiedener Leguminosengattungen, -arten und -sorten gezeigt. Es werden die Trennergebnisse der SDS-Elektrophorese mit Gelgradienten (T=4-22,5%) bzw. mit Gelen konstanter Porengröße (T=10%) verglichen. Das höchste Auslösungsvermögen und die beste Trennschärfe zeigen ultradünne Gradientengele. Die Nachweisempfindlichkeit ist bei allen Ultradünn-schicht-SDS-Elektrophoresen sehr hoch (0,1 g Protein/Bande). Da die auf Folie polymerisierten Gele sehr dünn sind (360 m) kann mit wesentlich verkürzten Trenn-, Färbe-, Entfärbe- und Trocknungszeiten gear-beitet werden. Es wird eine einfache Herstellung ultradünner Polyacrylamidgele mit exponentiellen konkaven Gradienten für die Trenndistanz von 10 cm mit einer Breite von 25 cm beschrieben. Die gerade und gleichmäßige Verteilung des Gradienten über die gesamte Gelbreite wird gezeigt. Mit der beschriebenen Methode werden bei den untersuchten Leguminosen-proteinen Molekulargewichte von 5 000 his 110 000 Dalton gefunden. Die Proteinmuster erweisen sich als gattungs- und artspezifisch.

     

Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus

Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single-chain F_v (scF_v), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scF_v were derived from three different human V_H germline genes. The binding specificities were confirmed by Western blots and ELISA. The scF_v preparations reacted with B. napus ENR, but not with β-keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scF_v 3.13 recognizes an epitope located within the n-terminal 80 amino acids of the enzyme molecule. The scF_v were used to detect ENR directly in extracts of B. napus seeds.     

Gene Variants Related to Cardiovascular and Pulmonary Diseases May Correlate with Severe Outcome of COVID-19

Background: Severe outcomes of COVID-19 account for up to 15% of all cases. The study aims to check if any gene variants related to cardiovascular (CVD) and pulmonary diseases (PD) are correlated with a severe outcome of COVID-19 in a Polish cohort of COVID-19 patients. Methods: In this study, a subset of 747 samples from unrelated individuals collected across Poland in 2020 and 2021 was used and whole-genome sequencing was performed. Results: The GWAS analysis of SNPs and short indels located in genes related to CVD identified one variant significant in COVID-19 severe outcome in the HADHA gene, while for the PD gene panel, we found two significant variants in the DRC1 gene. In this study, both potentially protective and risk variants were identified, of which variants in the HADHA gene deserve the most attention. Conclusions: This is the first study reporting the association between the HADHA and DRC1 genetic variants and COVID-19 severe outcome based on the cohort WGS analysis. Although all the identified variants are localised in introns, they may be correlated and therefore inherited along with other risk variants, potentially causative to severe outcome of COVID-19 but not discovered yet.     

MIL-100(Fe) Sub-Micrometric Capsules as a Dual Drug Delivery System

Nanoparticles of metal–organic frameworks (MOF NPs) are crystalline hybrid micro- or mesoporous nanomaterials that show great promise in biomedicine due to their significant drug loading ability and controlled release. Herein, we develop porous capsules from aggregate of nanoparticles of the iron carboxylate MIL-100(Fe) through a low-temperature spray-drying route. This enables the concomitant one-pot encapsulation of high loading of an antitumor drug, methotrexate, within the pores of the MOF NPs, and the collagenase enzyme (COL), inside the inter-particular mesoporous cavities, upon the formation of the capsule, enhancing tumor treatment. This association provides better control of the release of the active moieties, MTX and collagenase, in simulated body fluid conditions in comparison with the bare MOF NPs. In addition, the loaded MIL-100 capsules present, against the A-375 cancer cell line, selective toxicity nine times higher than for the normal HaCaT cells, suggesting that MTX@COL@MIL-100 capsules may have potential application in the selective treatment of cancer cells. We highlight that an appropriate level of collagenase activity remained after encapsulation using the spray dryer equipment. Therefore, this work describes a novel application of MOF-based capsules as a dual drug delivery system for cancer treatment.     

Development and Performance of Crystallization with Additives Applied on Different Milk Samples

Crystallization with additives is developed on milk samples from different processing treatments. Performance tests are carried out based on structure analysis of the crystallization patterns. Crystallization with milk as additive is applied following changes in milk after different processing treatments. When an aqueous cupric chloride dihydrate solution crystallizes in the presence of milk as additive, specific patterns emerge, which can be evaluated by image analysis. Milk samples were heated and homogenized in a pilot plant and characterized by various parameters. Furthermore, milk samples from the market were tested. Patterns from milk after heat treatment and homogenization are significantly different from those derived from untreated milk. The experiments could be reproduced for other milk samples, on different days and in another laboratory.     

Distribution and identification of culturable airborne microorganisms in a Swiss milk processing facility

Airborne communities (mainly bacteria) were sampled and characterized (concentration levels and diversity) at 1 outdoor and 6 indoor sites within a Swiss dairy production facility. Air samples were collected on 2 sampling dates in different seasons, one in February and one in July 2012 using impaction bioaerosol samplers. After cultivation, isolates were identified by mass spectrometry (matrix-assisted laser desorption/ionization-time-of-flight) and molecular (sequencing of 16S rRNA and rpoB genes) methods. In general, total airborne particle loads and total bacterial counts were higher in winter than in summer, but remained constant within each indoor sampling site at both sampling times (February and July). Bacterial numbers were generally very low (<100 cfu/m³ of air) during the different steps of milk powder production. Elevated bacterial concentrations (with mean values of 391 ± 142 and 179 ± 33 cfu/m³ of air during winter and summer sampling, respectively; n = 15) occurred mainly in the “logistics area,” where products in closed tins are packed in secondary packaging material and prepared for shipping. However, total bacterial counts at the outdoor site varied, with a 5- to 6-fold higher concentration observed in winter compared with summer. Twenty-five gram-positive and gram-negative genera were identified as part of the airborne microflora, with Bacillus and Staphylococcus being the most frequent genera identified. Overall, the culturable microflora community showed a composition typical and representative for the specific location. Bacterial counts were highly correlated with total airborne particles in the size range 1 to 5 µm, indicating that a simple surveillance system based upon counting of airborne particles could be implemented. The data generated in this study could be used to evaluate the effectiveness of the dairy plant’s sanitation program and to identify potential sources of airborne contamination, resulting in increased food safety.     

The effect of temperature and radiation on flavonol aglycones and flavonol glycosides of kale (Brassica oleracea var. sabellica)

The winter crop kale has a complex profile of different glycosylated and acylated flavonol glycosides which may be affected by global warming. To the best of our knowledge, compound–climate relationships for flavonol aglycones and flavonol glycosides were established for the first time. The investigated 10 major flavonol glycosides responded structure-dependent in the investigated temperature range between 0 and 12 °C and the photosynthetic active radiation range between 4 and 20 mol m⁻² d⁻¹, e.g. the decrease in temperature led to an increase in sinapic acid monoacylated and diacylated quercetin glycosides, while the sinapic acid monoacylated kaempferol glycosides showed a maximum at 4.5 °C. Furthermore, the hydroxycinnamic acid residues and the different number of glucose moieties in the 7-O position affected the response of kaempferol triglucosides. Consequently, global warming would result in lower concentrations of antioxidant-relevant quercetin glycosides in winter crops, suggesting a production at e.g. higher altitudes due to lower temperature.     

Rumination activity of dairy cows in the 24 hours before and after calving

Monitoring rumination behavior serves multiple purposes in feeding and herd management of dairy cows. The process of calving is a major event for cows, and a detailed understanding of alterations in behavioral patterns of animals in the time around calving is important in calving detection. The objective of this study was to describe the short-term changes in rumination patterns in dairy cows immediately before and after parturition. In total, 17 cows were fitted with rumination sensors that were able to monitor rumination time, number of rumination boli, and number of rumination jaw movements. Rumination time was decreased in the last 4 h antepartum and in the first 8 h postpartum. Cows stopped ruminating 123 ± 58 min (mean ± standard deviation) before calving and resumed ruminating 355 ± 194 min after calving. The number of rumination jaw movements and boli per day were decreased in the 24-h period postpartum. Rumination rate, the number of rumination jaw movements per rumination minute, and the number of boli per rumination minute changed little around calving. The calving event primarily influenced the duration and frequency of various rumination characteristics but not rumination intensity. Among detected characteristics, rumination time showed the greatest potential for monitoring of calving events.