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.     

Impact of compression,physical aging,and freezing rate on the crystallization characteristics of an amorphous sugar matrix

Amorphous matrices made up of sugar molecules, are frequently used in food and pharmaceutical industries. A drawback to their use is that they are susceptible to collapse, as a result of water uptake and an increase in temperature and subsequently crystallize. Herein, the crystallization characteristics of amorphous sugar (sucrose and α-lactose) preparations were analyzed, with the purpose of obtaining knowledge that could lead to the prediction of how long the amorphous state is retained under various conditions. The impact of compression, physical aging and freezing rate on the induction period (t_ind) for crystallization were examined. Freeze-dried sugar samples were compressed at 74 or 443 MPa (5 min) and then rehumidified at specified RHs. Some freeze-dried sucrose samples were physically aged, and alternatively freeze-drying was conducted under different conditions. The isothermal crystallization of the prepared samples at different temperatures (T), the glass transition and the crystallization temperature (T_cry) were measured, using differential scanning calorimetry. The compression markedly decreased the t_ind, while significantly lowered the hygroscopicity. Physical aging and slower-freezing also shortened the t_ind. The t_ind was found to be correlated exclusively with (T_cry–T), regardless of rehumidification, compression, sugar type, physical aging and freezing rate in the freeze-drying process.     

Phase transformation and interface segregation behavior in Si3N4 ceramics sintered with La2O3–Lu2O3 mixed additive

Microstructure and mechanical property of silicon nitride (Si₃N₄) ceramic are strongly dependent on the selection of sintering additives. When rare‐earth (RE) oxide is used as the sintering additive, segregation of RE ions at interface between Si₃N₄ grain and intergranular glassy film (IGF) is believed to play a critical role. Although the ionic radius of RE ion is known to be an empirical parameter to modify the mechanical property, the correlation between the segregated ions and their ionic radii is still under controversy. In order to address this issue, (i) rate of α‐β phase transformation and (ii) segregation behavior at the interface were studied for Si₃N₄ ceramics sintered using mixture of La₂O₃ and Lu₂O₃ as additives in this study. Specimens of Lu content 30% and higher exhibited lower activation energies for the α‐β phase transformation as compared with those of Lu content 20% and lower. In terms of the segregation behavior, La was preferably segregated at one site and Lu at the other site along β‐Si₃N₄/IGF interface in the specimens of Lu content 30% and higher. It is understood from these results that Lu segregation site should be more closely related with grain growth.     

Self-swarming robots that exploit hydrodynamical interaction

Although numerous swarm robotic systems have already been developed, they have exhibited limited adaptability. This was partly because the previous researchers designed the control schemes on the basis of informational interaction, without considering the physical effects of the environment. To tackle this problem, we employ an unconventional approach: we design a control scheme for swarm robots based on their physical interaction in a hydrodynamic field, inspired by biological swarming. Through simulations using a smoothed particle hydrodynamics method, we show that the proposed control scheme allows agents to form an ordered swarm in response to their environment.     

Die quenching limit of AA2024 aluminum alloy billet on servo press

Die quenching of AA2024 aluminum alloy billets was carried out on a servo press with ram-motion control of WC-20 mass%Co dies directly after solution heat treatment (SHT). To clarify the dependence on billet size for die quenching, two billets with a height of h₀ = 8 mm or 16 mm and with the same diameter of 16 mm were prepared. The cylindrical billets were heated in an electric furnace at 823 K and transferred to the press. Then the billets were uniaxially compressed with a reduction in height (Δh/h₀) of 2% or 5%, and further held between the dies. The sandwiching duration by dies (t_d.q.) was varied from 0 to 8 s. Based a measured temperature change, hardness and TG–DTA analysis, it is found that die quenching is successfully carried out without precipitation hardening only in the case of the billet with a height of 8 mm and t_d.q. > 6 s. The reduction in height is limited less than 5% by intergranular fracture on side surface of billet during the die quenching process.     

Characteristics of hydrogen-assisted intergranular fatigue crack growth in interstitial-free steel: role of plastic strain localization

The behavior of intergranular fatigue crack growth in an interstitial-free (IF) steel in a hydrogen environment was investigated at different frequencies. Focusing on the plastic strain localization, we observed details of the striation-like feature on the intergranular fracture surface, slip behavior around microvoids, and crystallographic orientation gradient underneath the fracture surface. It was determined that the intergranular fatigue crack growth mechanism in the IF steel is microvoid formation at the crack tip and subsequent coalescence with the crack. Moreover, it was found that the grain boundaries, acting as propagation paths, suffer from pre-damage arising from plastic strain localization near the grain boundaries even before the main crack propagates to a certain location. Therefore, fatigue cracks in a hydrogen environment easily propagate to the grain boundaries. The frequency dependence of fatigue crack growth in the hydrogen environment is significantly smaller than that in a low carbon steel, probably because of the frequency dependence of the pre-damage evolution behavior.     

Expression of Human DNAJ (Heat Shock Protein-40) B3 in Humanized UDP-glucuronosyltransferase 1 Mice

The human DNAJB3 gene encodes a DNAJ (Heat shock protein 40; Hsp40) homolog, subfamily B, member 3 chaperone protein (DNAJB3), which can be down-regulated in disease conditions, as observed in decreased expression of DNAJB3 mRNA in peripheral blood mononuclear cells (PBMC) of obese patients. Recently, humanized UDP-glucuronosyltransferase (UGT) 1 mice (hUGT1 mice) were developed, in which the introduced human UGT1 gene contained a gene encoding human DNAJB3. In the present study, we analyzed the expression of human DNAJB3 mRNA in hUGT1 mice. Among the examined tissues, the testis had the highest expression of human DNAJB3 mRNA, while the lowest expression was observed in the liver. We found that the pattern of tissue-specific expression of mouse Dnajb3 in hUGT1 mice was very similar to that of human DNAJB3. We further demonstrated that the expression of human DNAJB3 in the liver was significantly reduced in high-fat-diet-fed hUGT1 mice compared to the expression level in the control mice, indicating that the expression of human DNAJB3 in hUGT1 mice could be similarly regulated in disease conditions such as obesity. Humanized UGT1 mice might therefore be useful to investigate the physiological role of human DNAJB3 in vivo.     

Microencapsulation of bioactive compounds from mulberry (Morus alba L.) leaf extracts by protein–polysaccharide interactions

Mulberry leaf extracts were generated using four concentrations of ethanol (50%, 60%, 70%, and 95% v/v). A 60% ethanolic mulberry leaf extract (60E) yielded a high total phenolic content (TPC) and antioxidant activity using 1, 1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging and a ferric reducing antioxidant power (FRAP) assay. Coating materials were derived using a combination of soy protein isolates (SPI) and low methoxyl (LM) pectin in a 1:1 ratio. The effect of various parameters on microencapsulation, such as pH (3.5, 4.0, and 4.5) and the concentration of coating materials (2.5, 5.0 and 7.5% w/v), was studied. Microcapsules produced using 60E as a core material at pH 4.0 with 7.5% of coating material showed a high encapsulation yield, encapsulation efficiency, TPC and antioxidant activity.