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.     

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.     

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.     

Methods for determining pesticides and polychlorinated biphenyls in food samples--problems and challenges

Determination of residual amounts of pesticides and polychlorinated biphenyls (PCBs) in food samples requires the use of specific techniques regarding sample preparation as well as instrumental analysis which should be characterized by a very low detection limit. A problem associated with the use of pesticides and PCBs is the need for controlling their residues in the environment, particularly in food, as these chemicals show a propensity to accumulate. The analysis of food samples for the presence of pesticides and PCBs brings on many difficulties because of the specificity of sample preparation consisting of multistep purification procedures of samples that contain trace amounts of an analyte. Concentration determinations of pollutants that easily dissolve in complex matrices, particularly in the presence of a large apportionment of interfering substances, pose a big challenge. Therefore, the basic step in food analysis for the presence of pesticides and PCBs is sample preparation which mainly consists of analyte enrichment and the removal of interfering substances. But all steps of the analytical procedure that include sample collection and preparation, extraction of analytes from matrix, extract purification, and final determination, are very significant; their precision and correct application have a decisive effect on the final result.     

Multiphase visualisation of fat containing β-lactoglobulin-κ-carrageenan gels by confocal scanning laser microscopy, using a novel dye, V03-01136, for fat staining

In this study, the gel strength and visible microstructure of fat containing β-lactoglobulin-κ-carrageenan gels were investigated using puncture testing and confocal scanning laser microscopy, respectively. The gel strength was closely linked to the visible microstructure of the whey protein network as stained with Rhodamine B. Covalent labelling of κ-carrageenan with FITC prior to gel formation enabled localisation of the hydrocolloid phase, but caused a significant drop in the gel strength. This effect coincided with the observed reduction of the κ-carrageenan intrinsic viscosity, which was found to be a result of the labelling process. The use of a novel dye, V03-01136, for the staining of fat allowed for the specific and concurrent visualisation of the protein, hydrocolloid and fat phases under the conditions applied in the study.     

Phenotypic and PCR-based characterization of the microflora in Norvegia cheese during ripening

Microbiological sampling of Norvegia cheese from three cheese factories was done during ripening. The evolution of aerobic mesophilic bacteria, lactococci, lactobacilli, enterococci, presumptive leuconostoc and pediococci was investigated after 30, 90, 180 and 270 days of ripening. Isolates (135) of non-starter lactic acid bacteria (NSLAB) from nine Norvegia cheeses after 90, 180 and 270 days of ripening were examined. The isolates were tested by physiological and biochemical assays, species-specific PCR and 16S rDNA sequencing. After 90 days of ripening Leuconostoc spp., most probably from the starter, and the NSLAB specie Lactobacillus paracasei dominated among the isolates, however, after longer ripening Lb. paracasei dominated. The development and evolution of the microflora in Norvegia varied according to dairy and ripening time.     

Influence of dynamic stimulation,visual perception and individual susceptibility to car sickness during controlled stop-and-go driving

Forschung im Ingenieurwesen - The enjoyment and usage of highly automated vehicles might be reduced by motion sickness. This study addresses the influencing factors of dynamic stimulation, visual...