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.     

Correlation between motion of source bender element and feedback signal induced by self-monitoring

Since the introduction of bender element tests to soil testing, the reliability of the estimated travel time has been the most serious problem. The author has previously shown a potential solution whereby removing the response of the bender element subsystem from the whole response could dramatically improve the accuracy of the travel time estimation. In order to lay the foundation for estimating the response of the bender element subsystem, this paper examines the correlation between the displacements of the element and the induced feedback signals by employing a self-monitoring element. The response of the self-monitoring element is modeled as a transfer function involving two internal transfer functions that relate the input signals to the displacements and the displacements to the feedback signals, respectively. Using a laser displacement sensor, the displacements are directly measured through the entire surface and reveal the three-dimensional bending motion of the element oscillating in both longitudinal and width directions. The feedback signals are similar to, but inconsistent with, the tip displacements, and an attempt is made to correct the feedback signals. Finally, a conclusion is given on the potential for estimating the response of the bender element subsystem using the self-monitoring element.     

In Vivo Electrophysiology of Peptidergic Neurons in Deep Layers of the Lumbar Spinal Cord after Optogenetic Stimulation of Hypothalamic Paraventricular Oxytocin Neurons in Rats

The spinal ejaculation generator (SEG) is located in the central gray (lamina X) of the rat lumbar spinal cord and plays a pivotal role in the ejaculatory reflex. We recently reported that SEG neurons express the oxytocin receptor and are activated by oxytocin projections from the paraventricular nucleus of hypothalamus (PVH). However, it is unknown whether the SEG responds to oxytocin in vivo. In this study, we analyzed the characteristics of the brain–spinal cord neural circuit that controls male sexual function using a newly developed in vivo electrophysiological technique. Optogenetic stimulation of the PVH of rats expressing channel rhodopsin under the oxytocin receptor promoter increased the spontaneous firing of most lamina X SEG neurons. This is the first demonstration of the in vivo electrical response from the deeper (lamina X) neurons in the spinal cord. Furthermore, we succeeded in the in vivo whole-cell recordings of lamina X neurons. In vivo whole-cell recordings may reveal the features of lamina X SEG neurons, including differences in neurotransmitters and response to stimulation. Taken together, these results suggest that in vivo electrophysiological stimulation can elucidate the neurophysiological response of a variety of spinal neurons during male sexual behavior.     

Adaptation to climate change and conservation of biodiversity using green infrastructure

In recent years, we have experienced mega‐flood disasters in Japan due to climate change. In the last century, we have been building disaster prevention infrastructure (artificial levees and dams, referred to as “grey infrastructure”) to protect human lives and assets from floods, but these hard protective measures will not function against mega‐floods. Moreover, in a drastically depopulating society such as that in Japan, farmland abandonment prevails, and it will be more difficult to maintain grey infrastructure with a limited tax income. In this study, we propose the introduction of green infrastructure (GI) as an adaptation strategy for climate change. If we can use abandoned farmlands as GI, they may function to reduce disaster risks and provide habitats for various organisms that are adapted to wetland environments. First, we present a conceptual framework for disaster prevention using a hybrid of GI and conventional grey infrastructure. In this combination, the fundamental GI, composed of forests and wetlands in the catchment (GI‐1) and additional multilevel GIs such as flood control basins that function when floodwater exceeds the planning level (GI‐2) are introduced. We evaluated the flood attenuation function (GI‐1) of the Kushiro Wetland using a hydrological model and developed a methodology for selecting suitable locations of GI‐2, considering flood risk, biodiversity and the distribution of abandoned farmlands, which represent social and economic costs. The results indicated that the Kushiro Wetland acts as a large natural reservoir that attenuates the hydrological peak discharge during floods and suitable locations for introducing GI‐2 are concentrated in floodplain areas developing in the downstream reaches of large rivers. Finally, we discussed the network structure of GI‐1 as a hub and GI‐2 as a dispersal site for conservation of the Red‐crowned Crane, one of the symbolic species of Japan.     

New amphoteric surfactants derived from lysine. I. Preparation and properties of Nε-acyllysine derivatives-acyllysine derivatives

New amphoteric surfactants were prepared from N^ε-acyllysine which was obtained by the thermal dehydration of a higher fatty acid salt of lysine and was not soluble in water. N^α,N^α-dimethyl-N^ε-acyllysine was prepared by the catalytic reductive condensation of N^ε-acyllysine ester with formaldehyde in good yield. N^α,N^α,N^α-trimethyl-N^ε-acyllysine was obtained from the reaction of N^α,N^α-dimethyl-N^ε-acyllysine ester with methyl iodide. Confirmation of the structure of these derivatives was obtained by spectrometric and spectroscopic analyses. The solubility of N^ε-acyllysine was improved significantly by the introduction of N^α-methyl groups. Physicochemical and surface active properties of the derivatives were investigated in terms of isoelectric points, dissolution temperatures, surface tensions, critical micelle concentrations (cmc), foaming properties and wetting powers. N^α,N^α,N^α-trimethyl-N^ε-acyllysine had lower dissolution temperatures than N^α,N^α-dimethyl-N^ε-acyllysine. The latter showed lower surface tensions than the former at cmc. N^α,N^α-dimethyl-N^ε-lauroyllysine was best in wetting power and foaming property.     

The effect of a nucleotide-nucleoside solution on hepatic regeneration after partial hepatectomy in rats

After hepatectomy, purine and pyrimidine metabolism is a key process in the synthesis of DNA and RNA and maintaining cellular energy metabolism. The purpose of this study is to evaluate changes in blood purine and pyrimidine levels after partial hepatectomy and the effect of purine and pyrimidine nucleoside solution injection on hepatic regeneration under the hypothesis that the rat after partial hepatectomy requires substrates for salvage nucleotide synthesis and changes blood nucleoside and nucleobase levels. Blood levels of nucleotides, nucleosides, and nucleobase by high-performance liquid chromatography method and liver ATP level by enzymatic analysis, and the effect of preoperative injection of nucleoside solution (OG-VI) on hepatic regeneration ratio and hepatocytes DNA synthesis, were assessed in rats after 70% partial hepatectomy. Decreased liver adenosine triphosphate and increased plasma xanthine and hypoxanthine after partial hepatectomy indicated an increase in catabolism of purine nucleotides in regenerating liver. Plasma thymidine and cytidine levels increased, then returned to the prevalue, suggesting that the thymidine and cytidine pool was enlarged. OG-VI increased labeling indices of hepatocytes at postoperative d 1 (POD) and hepatic regeneration ratio at POD 14. Blood purine nucleobase and pyrimidine nucleoside levels change after partial hepatectomy and preoperative supply of nucleoside solution is effective for increasing hepatocytes DNA synthesis and hepatic regeneration after partial hepatectomy.     

Intelligent decision support systems for nuclear power plants in Japan

This paper describes the MITI (Ministry of International Trade and Industry) projects for decision support systems for NPPs in Japan. Main attention is paid to the new project of an advanced man-machine system for nuclear power plants, of which a conceptual design was initiated in 1984. Some of the aspects of the design and the outline of the prototype system are discussed.