Production of Recombinant Single-Chain Eel Luteinizing Hormone and Follicle-Stimulating Hormone Analogs in Chinese Hamster Ovary Suspension Cell Culture.

We produced rec-single chain eel luteinizing (rec-eel LH) and follicle-stimulating (rec- eel FSH) hormones displaying high biological activity in Chinese hamster ovary suspension (CHO-S) cells. We constructed several mutants, in which a linker, including an O-linked glycosylated carboxyl-terminal peptide (CTP) of an equine chorionic gonadotropin (eCG) ß-subunit, was attached between the ß- and α-subunit (LH-M and FSH-M) or in the N-terminal (C-LH and C-FSH) or C-terminal (LH-C and FSH-C) regions. The plasmids were transfected into CHO-S cells, and culture supernatants were collected. The secretion of mutants from the CHO-S cells was faster than that of eel LHß/α-wt and FSHß/α-wt proteins. The molecular weight of eel LHß/α-wt and eel FSHß/α-wt was 32-34 and 34-36 kDa, respectively, and that of LH-M and FSH-M was 40-43 and 42-45 kDa, respectively. Peptide-N-glycanase F-treatment markedly decreased the molecular weight by approximately 8-10 kDa. The EC50 value and the maximal responsiveness of the eel LH-M and eel FSH-M increased compared with the wild-type proteins. These results show that the CTP region plays a pivotal role in early secretion and signal transduction. We suggest that novel rec-eel LH and FSH proteins, exhibiting potent activity, could be produced in large quantities using a stable CHO cell system.

Thin Film Composite Membranes as a New Category of Alkaline Water Electrolysis Membranes.

Alkaline water electrolysis (AWE) is considered a promising technology for green hydrogen (H2 ) production. Conventional diaphragm-type porous membranes have a high risk of explosion owing to their high gas crossover, while nonporous anion exchange membranes lack mechanical and thermochemical stability, limiting their practical application. Herein, a thin film composite (TFC) membrane is proposed as a new category of AWE membranes. The TFC membrane consists of an ultrathin quaternary ammonium (QA) selective layer formed via Menshutkin reaction-based interfacial polymerization on a porous polyethylene (PE) support. The dense, alkaline-stable, and highly anion-conductive QA layer prevents gas crossover while promoting anion transport. The PE support reinforces the mechanical and thermochemical properties, while its highly porous and thin structure reduces mass transport resistance across the TFC membrane. Consequently, the TFC membrane exhibits unprecedentedly high AWE performance (1.16 A cm-2 at 1.8 V) using nonprecious group metal electrodes with a potassium hydroxide (25 wt%) aqueous solution at 80 °C, significantly outperforming commercial and other lab-made AWE membranes. Moreover, the TFC membrane demonstrates remarkably low gas crossover, long-term stability, and stack cell operability, thereby ensuring its commercial viability for green H2 production. This strategy provides an advanced material platform for energy and environmental applications.

Effect of Bacillus sp. Supplementation Diet on Survival Rate and Microbiota Composition in Artificially Produced Eel Larvae (Anguilla japonica).

This study was performed to investigate the effect of microbial supplementation diet on the survival rate and microbiota composition of artificially produced eel larvae. Microorganisms supplemented in the diet were isolated from wild glass eel intestines and identified as Bacillus sp. through 16S rRNA sequencing analysis. In vitro tests confirmed that the strain had no hemolytic activity and virulence genes. Microbial supplemental feeding significantly increased the survival rate of artificially produced eel larvae for 30 days post-hatchling compared with that of the control group. It also caused changes in the α-diversity, ß-diversity, and relative abundance of the bacterial communities. Analysis via phylogenetic investigation of communities by reconstruction of unobserved states predicted that these microbial community changes would significantly increase the carbohydrate metabolism, membrane transport, and cellular community pathway of the microbial supplementation group. Therefore, microbial supplementation feeding for eel aquaculture could increase the viability of artificially produced eel larvae and alter the microbial composition to induce metabolic changes.

Morphological and Allometric Changes in Anguilla japonica Larvae.

The freshwater eel Anguilla japonica is rapidly decreasing in number and has not yet been successfully mass produced. This may be at least partially attributable to the unique and long early life history of the eel. Therefore, we investigated its ontogeny of morphometry and growth pattern in larval stages to provide baseline information for understanding the early life history and improving seed rearing technology. This study was conducted for 200 days after hatching (DAH) and analyzed morphometry and allometry for eel larvae. The following cultured eel larval stages were identified: the yolk sac larvae stage (0-6 DAH, 3.23-6.85 mm total length (TL)), the pre-leptocephalus stage (7-30 DAH, 6.85-15.31 mm TL), and the leptocephalus stage (50-200 DAH, 15.31-60.06 mm TL). Cultured and wild eel larvae could be divided into characteristic larval stages at similar sizes. However, compared to wild eels, cultured eels had a slower growth rate and fewer preanal myomeres. Meanwhile, cultured eel larvae rarely had a mixed feeding period as the absorption of endogenous reserves was completed by 7 DAH. The lower jaw of eel larvae was significantly longer than the upper jaw from 50 DAH. In the pre-leptocephalus and leptocephalus stages, eel larvae showed continuous positive allometric growth at trunk height and tail muscle height with change to the willow leaf-like form. These growth characteristics may be the result of adaptation to the migration over long distances and to a diel vertical migration. The inflection point in the body parts growth patterns showed only before 30 DAH, and mass mortality appeared at this period. Therefore, to improve the growth and survival rates of cultured eel seed, it is necessary to focus on improving the feeding and rearing protocol until 30 DAH.

Highly Selective and pH-Stable Reverse Osmosis Membranes Prepared via Layered Interfacial Polymerization.

Ultrathin and smooth polyamide (PA) reverse osmosis (RO) membranes have attracted significant interest due to their potential advantages of high permeance and low fouling propensity. Although a layered interfacial polymerization (LIP) technique aided by the insertion of a polyelectrolyte interlayer has proven effective in fabricating ultrathin and uniform membranes, the RO performance and pH stability of the fabricated LIP membrane remain inadequate. In this study, a poly(piperazineamide) (PIPA) layer prepared via interfacial polymerization (IP) was employed as an interlayer to overcome the limitations of the prototype LIP method. Similar to the control polyelectrolyte-interlayered LIP membrane, the PIPA-interlayered LIP (pLIP) membrane had a much thinner (~20 nm) and smoother selective layer than the membrane fabricated via conventional IP due to the highly surface-confined and uniform LIP reaction. The pLIP membrane also exhibited RO performance exceeding that of the control LIP and conventional IP-assembled membranes, by enabling denser monomer deposition and a more confined interfacial reaction. Importantly, the chemically crosslinked PIPA interlayer endowed the pLIP membrane with higher pH stability than the control polyelectrolyte interlayer. The proposed strategy enables the fabrication of high-performance and pH-stable PA membranes using hydrophilic supports, which can be applied to other separation processes, including osmosis-driven separation and organic solvent filtration.

Effects of Capping Layers with Different Metals on Electrical Performance and Stability of p-Channel SnO Thin-Film Transistors.

In this study, the effects of capping layers with different metals on the electrical performance and stability of p-channel SnO thin-film transistors (TFTs) were examined. Ni- or Pt-capped SnO TFTs exhibit a higher field-effect mobility (µFE), a lower subthreshold swing (SS), a positively shifted threshold voltage (VTH), and an improved negative-gate-bias-stress (NGBS) stability, as compared to pristine TFTs. In contrast, Al-capped SnO TFTs exhibit a lower µFE, higher SS, negatively shifted VTH, and degraded NGBS stability, as compared to pristine TFTs. No significant difference was observed between the electrical performance of the Cr-capped SnO TFT and that of the pristine SnO TFT. The obtained results were primarily explained based on the change in the back-channel potential of the SnO TFT that was caused by the difference in work functions between the SnO and various metals. This study shows that capping layers with different metals can be practically employed to modulate the electrical characteristics of p-channel SnO TFTs.

Floating Ni Capping for High-Mobility p-Channel SnO Thin-Film Transistors.

We utilized Ni as a floating capping layer in p-channel SnO thin-film transistors (TFTs) to improve their electrical performances. By utilizing the Ni as a floating capping layer, the p-channel SnO TFT showed enhanced mobility as high as 10.5 cm2·V-1·s-1. The increase in mobility was more significant as the length of Ni capping layer increased and the thickness of SnO active layer decreased. The observed phenomenon was possibly attributed to the changed vertical electric field distribution and increased hole concentration in the SnO channel by the floating Ni capping layer. Our experimental results demonstrate that incorporating the floating Ni capping layer on the channel layer is an effective method for increasing the field-effect mobility in p-channel SnO TFTs.

Oxide Thin-Film Transistor-Based Vertically Stacked Complementary Inverter for Logic and Photo-Sensor Operations.

Numerous studies have addressed the utilization of oxide thin-film transistor (TFT)-based complementary logic circuits that are based on two-dimensional (2D) planar structures. However, there are fundamental limits to the 2D planar structured complementary logic circuits, such as a large dimension and a large parasitic resistance. This work demonstrated a vertically stacked three-dimensional complementary inverter composed of a p-channel tin monoxide (SnO) TFT and an n-channel indium-gallium-zinc oxide (IGZO) TFT. A bottom-gate p-channel SnO TFT was formed on the top-gate n-channel IGZO TFT with a shared common gate electrode. The fabricated vertically stacked complementary inverter exhibited full swing characteristics with a voltage gain of ~33.6, a high noise margin of 3.13 V, and a low noise margin of 3.16 V at a supplied voltage of 10 V. The achieved voltage gain of the fabricated complementary inverter was higher than that of the vertically stacked complementary inverters composed of other oxide TFTs in previous works. In addition, we showed that the vertically stacked complementary inverter exhibited excellent visible-light photoresponse. This indicates that the oxide TFT-based vertically stacked complementary inverter can be used as a sensitive photo-sensor operating in the visible spectral range with the voltage read-out scheme.

Polyethylene Battery Separator as a Porous Support for Thin Film Composite Organic Solvent Nanofiltration Membranes.

Organic solvent nanofiltration (OSN) has made significant advances recently, and it is now possible to fabricate thin film composite (TFC) membranes with a selective layer thickness below 10 nm that gives ultrafast solvent permeance. However, such high permeance is inadvertently limited by the support membrane beneath the selective layer, and thus there is an urgent need to develop a suitable support to maximize TFC performance. In this work, we employed a commercially available polyethylene (PE) battery separator as a porous support to fabricate high performance TFC OSN membranes. To deposit a uniform polyamide selective layer onto the porous support via interfacial polymerization, the PE support was hydrophilized with O2 plasma and the reaction efficiency was optimized using a surfactant. Owing to the high surface porosity of the PE support and the high permselectivity of the PA layer, the PE-supported TFC membrane outperformed the previously reported OSN membranes and its performance exceeded the current performance upper bound. A solvent activation step dramatically improved the solvent permeance by 5-fold while maintaining nanoseparation properties. In addition to the superior OSN performance, the commercial availability of the PE support and simplified TFC fabrication protocol would make the PE-supported OSN membranes commercially attractive.

Eleutherococcus senticosus extract attenuates LPS-induced iNOS expression through the inhibition of Akt and JNK pathways in murine macrophage.

Eleutherococcus senticosus (Araliaceae) is immunological modulator which has been successfully used for anti-inflammatory effectors on anti-rheumatic diseases in oriental medicine. Mitogen-activated protein kinases (MAPKs) and Akt modulate the transcription of many genes involved in the inflammatory process. In this study, we investigated the inhibitory effects of Eleutherococcus senticosus on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharides (LPS)-activated macrophages. Finally, we studied the involvement of MAPKs and Akt signaling in the protective effect of Eleutherococcus senticosus in LPS-activated macrophages. Eleutherococcus senticosus significantly attenuated LPS-induced iNOS expression but not COX-2 expression. In using the standard inhibitors (MAPKs and Akt), our results show that Eleutherococcus senticosus downregulates inflammatory iNOS expression by blocking JNK and Akt activation.