Accelerated Discovery of Potent Bioactive anti-TNFα Aptamers by Microbead-Assisted Capillary Electrophoresis (MACE)-SELEX

Dysregulation of tumor necrosis factor-α (TNFα), a pro-inflammatory cytokine, causes several diseases, making it an important therapeutic target. Here, we identified a novel DNA aptamer against human TNFα using in vitro selection, which included a high exclusion pressure process against non-binding and weak binders through microbead-assisted capillary electrophoresis (MACE) in only three rounds. Among the 15 most enriched aptamers, Apt14 exhibited the highest inhibitory activity for the interaction between TNFα and its cognate receptor in mouse L929 cells. For further improving the bioactivity of the aptamer, dimerization programed by hybridization was evaluated, resulting in the Apt14 dimer exhibited a twofold higher binding affinity and stronger inhibition compared to the monomer counterpart. Rapid identification of bioactive aptamers using MACE in combination with facile dimerization by hybridization accelerates the discovery of novel bioactive aptamers, paving the way toward replacing current monoclonal antibody therapy with the less expensive and non-immunogenic aptamer therapy.     

Reversible Fragmentation and Self‐Assembling of Nematic Liquid Crystal Droplets on Functionalized Pyroelectric Substrates

Investigation on the behavior of nematic liquid crystals on functionalized polar dielectric crystal substrates is accomplished. Very interesting effects can be observed in maneuvering liquid crystal droplets on the substrate surface, driven by electric fields generated by pyroelectric effect. Reversible drops fragmentation and self‐assembling in different configurations can be achieved. The dynamics of the observed phenomena is studied and the repeatability of the process is full assessed.     

UHT milk contains multiple forms of αS1-casein that undergo degradative changes during storage

Milk proteins are susceptible to chemical changes during processing and storage. We used proteomic tools to analyse bovine α_S1-casein in UHT milk. 2-D gels of freshly processed milk α_S1-casein was presented as five or more spots due to genetic polymorphism and variable phosphorylation. MS analysis after phosphopeptide enrichment allowed discrimination between phosphorylation states and genetic variants. We identified a new alternatively-spliced isoform with a deletion of exon 17, producing a new C-terminal sequence, K¹⁶⁴SQVNSEGLHSYGL¹⁷⁷, with a novel phosphorylation site at S¹⁷⁴. Storage of UHT milk at elevated temperatures produced additional, more acidic α_S1-casein spots on the gels and decreased the resolution of minor forms. MS analysis indicated that non-enzymatic deamidation and loss of the N-terminal dipeptide were the major contributors to the changing spot pattern. These results highlight the important role of storage temperature in the stability of milk proteins and the utility of proteomic techniques for analysis of proteins in food.     

The study on the properties of TiCxN1−x coatings processed by cathodic arc physical vapour deposition

Titanium carbonitride (TiCN) is a popular hard coating for carbide cutting tools in various applications. The properties of TiCN are within its composition and can be controlled by maintaining the C–N ratio within the coating to a certain level. This paper studied the influence of carbon content and coating composition within TiC_xN_1?x coatings with regard to their mechanical properties. The substrate used was tungsten carbide (WC-6Co), which was prepared in-house through a powder metallurgy process, while the TiC_xN_1?x coatings were deposited in-house using cathodic arc physical vapour deposition (CAPVD). TiC_xN_1?x coatings improved the mechanical properties of carbide inserts. An increase in carbon content within TiC_xN_1?x coatings improved surface lubricity, reduced coefficient of friction, improved surface microhardness and increased Young's modulus, but reduced thermal conductivity of carbide inserts. The colour of TiC_xN_1?x coatings also changed with carbon content.     

Distribution of Salmonella clonal groups in four Brazilian feed mills

Feed is one of the major vehicles for Salmonella transmission to pigs, and its spread during the milling process may play an important role in feed contamination. Therefore, a cross-sectional study was carried out in four feed mills in order to evaluate the frequency of Salmonella isolation at various stages of feed production and to track the spread of genotypically related isolates grouped by pulsed-field gel electrophoresis (PFGE). From a total of 1269 analyzed samples, 63 (4.96%) showed the presence of Salmonella. Evaluated feed mills (A, B, C and D) presented 3.5% (n = 11/317), 1.7% (n = 5/289), 7.5% (n = 23/308) and 7.0% (n = 25/355) positive samples, respectively. Twenty-three serovars were identified, with the most frequently detected being Montevideo (n = 14, 22.2%), Anatum (n = 8, 12.7%) and Senftenberg (n = 8, 12.7%). The isolation of Salmonella was significantly higher (p = 0.002) in samples with the presence of total coliforms (36/489; 7.36%) than in the coliform-negative samples (27/780; 3.46%). Conveyors (OR = 4.43, 95% CI: 2.43–8.09) were the most likely sites of Salmonella isolation, followed by dust settled on the feed mill's floor (OR = 2.88, 95% CI: 1.41–5.33). Isolates indistinguishable on PFGE or belonging to pulsotypes with a high similarity (>95%) were identified in serovars Agona, Infantis, Montevideo, Orion, Senftenberg and Worthington. In particular, clonal groups of serovars Montevideo and Senftenberg were found to be disseminated among different sample types (ingredients, dust collected from the premise's floor and complete feed) or to be endemic in the feed mills. The dissemination of Salmonella clonal groups demonstrates the importance of control measures to avoid dust and debris accumulation on equipment surfaces.     

Microbial Diversity of Ground Beef Products in South Korean Retail Market Analyzed by PCR-DGGE and 454 Pyrosequencing

The purpose of this study was to assess the microbial community in ground beef purchased from local stores in the north central area of South Korea. A total of 31 ground beef samples showed 3.77 ± 5.16 × 10⁵ CFU/g on aerobic plate counts, and PCR-based denaturing gradient gel electrophoresis was performed to assess the microbial community, which revealed a diverse profile among the products. Four of these samples were selected for further analyses via 454 pyrosequencing. Pseudomonas was the most frequently identified genus in terms of relative abundance (4.54–73.8%), and other bacteria such as Lactobacillus, Acinetobacter, and Carnobacterium were also identified in the ground beef. The presence of bacteria can affect the spoilage of ground beef products depending on the microbial populations, and understanding these populations may help limit spoilage and/or foodborne illness. This study illustrates the microbial diversity occurring among Korean ground beef products, even within a local region, and provides a molecular approach for potential ground beef microbial quality assessment.