Fluorescence-Polarization-Based Assaying of Lysozyme with Chitooligosaccharide Tracers.

Lysozyme is a well-known enzyme found in many biological fluids which plays an important role in the antibacterial protection of humans and animals. Lysozyme assays are used for the diagnosis of a number of diseases and utilized in immunohistochemistry, genetic and cellular engineering studies. The assaying methods are divided into two categories measuring either the concentration of lysozyme as a protein or its activity as an enzyme. While the first category of methods traditionally uses an enzyme-linked immunosorbent assay (ELISA), the methods for the determination of the enzymatic activity of lysozyme use either live bacteria, which is rather inconvenient, or natural peptidoglycans of high heterogeneity and variability, which leads to the low reproducibility of the assay results. In this work, we propose the use of a chemically synthesized substrate of a strictly defined structure to measure in a single experiment both the concentration of lysozyme as a protein and its enzymatic activity by means of the fluorescence polarization (FP) method. Chito-oligosaccharides of different chain lengths were fluorescently labeled and tested leading to the selection of the pentasaccharide as the optimal size tracer and the further optimization of the assay conditions for the accurate (detection limit 0.3 µM) and rapid (<30 min) determination of human lysozyme. The proposed protocol was applied to assay human lysozyme in tear samples and resulted in good correlation with the reference assay. The use of synthetic fluorescently labeled tracer, in contrast to natural peptidoglycan, in FP analysis allows for the development of a reproducible method for the determination of lysozyme activity.

ASCA-related antibodies in the blood sera of healthy donors and patients with colorectal cancer: characterization with oligosaccharides related to Saccharomyces cerevisiae mannan.

Mannans are polysaccharide antigens expressed on the cell wall of different fungal species including Saccharomyces cerevisiae and Candida spp. These fungi are components of the normal intestinal microflora, and the presence of antibodies to fungal antigens is known to reflect the features of the patient's immune system. Thus, titers of IgG and IgA antibodies against Saccharomyces cerevisiae mannan (ASCA) are markers for clinical diagnostics of inflammatory bowel diseases. The complex organization and heterogeneity of cell-wall mannans may reduce the quality and reproducibility of ELISA results due to interference by different antigenic epitopes. In this research, we analyzed the levels of IgG antibodies in the sera of healthy donors and patients with colorectal cancer using an array of synthetic oligosaccharides related to distinct fragments of fungal mannan. This study aimed to establish the influence of oligosaccharide structure on their antigenicity. Variations in the structure of the previously established ASCA epitope (changing type of linkage, chain length, and the presence of branches) significantly modified the ability of ligands to bind to circulating antibodies in blood sera. The study showed that surface presentation density of the ligand critically affects the results of enzyme immunoassay. The transition from natural coating antigens to their corresponding synthetic mimetics with a defined structure opens new opportunities for improving existing ELISA test systems, as well as developing diagnostic kits with new properties.

Sodium Hypophosphite as a Bulk and Environmentally Friendly Reducing Agent in the Reductive Amination.

NaH2PO2 was found to promote reductive amination. Being nontoxic, stable, environmentally benign, and available in bulk amounts, this reducing agent showed a powerful potential to compete with classical reductants applied in the target process. An E factor of 1 was achieved for the substrate scope. Different carbonyl compounds reacted with amines under the developed conditions. The reaction demonstrated a great compatibility with a wide range of functional groups. Reaction conditions were scaled up to 200-fold.

Ediacara biota flourished in oligotrophic and bacterially dominated marine environments across Baltica.

Middle-to-late Ediacaran (575-541 Ma) marine sedimentary rocks record the first appearance of macroscopic, multicellular body fossils, yet little is known about the environments and food sources that sustained this enigmatic fauna. Here, we perform a lipid biomarker and stable isotope (δ15Ntotal and δ13CTOC) investigation of exceptionally immature late Ediacaran strata (<560 Ma) from multiple locations across Baltica. Our results show that the biomarker assemblages encompass an exceptionally wide range of hopane/sterane ratios (1.6-119), which is a broad measure of bacterial/eukaryotic source organism inputs. These include some unusually high hopane/sterane ratios (22-119), particularly during the peak in diversity and abundance of the Ediacara biota. A high contribution of bacteria to the overall low productivity may have bolstered a microbial loop, locally sustaining dissolved organic matter as an important organic nutrient. These oligotrophic, shallow-marine conditions extended over hundreds of kilometers across Baltica and persisted for more than 10 million years.

Formalizing Human-Robot Mutual Adaptation: A Bounded Memory Model.

Mutual adaptation is critical for effective team collaboration. This paper presents a formalism for human-robot mutual adaptation in collaborative tasks. We propose the bounded-memory adaptation model (BAM), which captures human adaptive behaviors based on a bounded memory assumption. We integrate BAM into a partially observable stochastic model, which enables robot adaptation to the human. When the human is adaptive, the robot will guide the human towards a new, optimal collaborative strategy unknown to the human in advance. When the human is not willing to change their strategy, the robot adapts to the human in order to retain human trust. Human subject experiments indicate that the proposed formalism can significantly improve the effectiveness of human-robot teams, while human subject ratings on the robot performance and trust are comparable to those achieved by cross training, a state-of-the-art human-robot team training practice.