Complete genome sequence of a tentative novel capillovirus isolated from Gerbera jamesonii.

The currently named gerbera virus A (GeVA) has been shown to be a novel capillovirus with a complete genome of 6929 nucleotides (nt) (GenBank accession no. OM525829.1). GeVA was detected in Gerbera jamesonii using high-throughput RNA sequencing analysis. The GeVA genome is a single linear RNA with two open reading frames (ORF), similar to those of other capilloviruses. The larger ORF encodes a polyprotein containing four domains, while the smaller ORF encodes a movement protein. The complete genome had 41.0-54.9% nt sequence identity to other those of capilloviruses, while the polyprotein and the movement protein had 26.5-36.4% and 13.1-32.2% amino acid (aa) sequence identity, respectively. Two UUAGGU promoters for subgenomic RNA (sgRNA) transcription were also identified in this study. BLAST analysis demonstrated that the GeVA genome shared the highest sequence similarity with rubber tree capillovirus 1 (MN047299.1) (complete nucleotide sequence identity, 68.54%; polyprotein amino acid sequence identity, 44.53%). Phylogenetic analysis based on complete genome and replication protein sequences placed GeVA alongside other members of the genus Capillovirus in the family Betaflexiviridae. These data suggest that GeVA is a new member of the genus Capillovirus.

MAML1/2 promote YAP/TAZ nuclear localization and tumorigenesis.

The Hippo pathway plays a pivotal role in tissue homeostasis and tumor suppression. YAP and TAZ are downstream effectors of the Hippo pathway, and their activities are tightly suppressed by phosphorylation-dependent cytoplasmic retention. However, the molecular mechanisms governing YAP/TAZ nuclear localization have not been fully elucidated. Here, we report that Mastermind-like 1 and 2 (MAML1/2) are indispensable for YAP/TAZ nuclear localization and transcriptional activities. Ectopic expression or depletion of MAML1/2 induces nuclear translocation or cytoplasmic retention of YAP/TAZ, respectively. Additionally, mutation of the MAML nuclear localization signal, as well as its YAP/TAZ interacting region, both abolish nuclear localization and transcriptional activity of YAP/TAZ. Importantly, we demonstrate that the level of MAML1 messenger RNA (mRNA) is regulated by microRNA-30c (miR-30c) in a cell-density-dependent manner. In vivo and clinical results suggest that MAML potentiates YAP/TAZ oncogenic function and positively correlates with YAP/TAZ activation in human cancer patients, suggesting pathological relevance in the context of cancer development. Overall, our study not only provides mechanistic insight into the regulation of YAP/TAZ subcellular localization, but it also strongly suggests that the miR30c-MAML-YAP/TAZ axis is a potential therapeutic target for developing novel cancer treatments.

Fabrication and Evaluation of Embroidery-Based Electrode for EMG Smart Wear Using Moss Stitch Technique.

Wearable 2.0 research has been conducted on the manufacture of smart fitness wear that collects bio-signals through the wearing of a textile-based electrode. Among them, the electromyography (EMG) suit measures the electrical signals generated by the muscles to check their activity, such as contraction and relaxation. General gel-type electrodes have been reported to cause skin diseases due to an uncomfortable feel and skin irritation when attached to the skin for a long time. Dry electrodes of various materials are being developed to solve this problem. Previous research has reported EMG detectio performance and conducted economic comparisons according to the size and shape of the embroidery electrode. On the other hand, these embroidery electrodes still have foreign body sensations. In this study, a moss sEMG electrode was produced with various shapes (W3 and WF) and loop lengths (1-5 mm). The optimized conditions of the embroidery-based electrodes were derived and analyzed with the tactile comfort factors and sensing performances. As the loop length of the electrode increased, MIU and Qmax increased, but the SMD decreased due to the free movement of the threads constituting the loop. Impedance and sEMG detection performance showed different trends depending on the electrode type.

LDL receptor-related protein LRP6 senses nutrient levels and regulates Hippo signaling.

Controlled cell growth and proliferation are essential for tissue homeostasis and development. Wnt and Hippo signaling are well known as positive and negative regulators of cell proliferation, respectively. The regulation of Hippo signaling by the Wnt pathway has been shown, but how and which components of Wnt signaling are involved in the activation of Hippo signaling during nutrient starvation are unknown. Here, we report that a reduction in the level of low-density lipoprotein receptor-related protein 6 (LRP6) during nutrient starvation induces phosphorylation and cytoplasmic localization of YAP, inhibiting YAP-dependent transcription. Phosphorylation of YAP via loss of LRP6 is mediated by large tumor suppressor kinases 1/2 (LATS1/2) and Merlin. We found that O-GlcNAcylation of LRP6 was reduced, and the overall amount of LRP6 was decreased via endocytosis-mediated lysosomal degradation during nutrient starvation. Merlin binds to LRP6; when LRP6 is less O-GlcNAcylated, Merlin dissociates from it and becomes capable of interacting with LATS1 to induce phosphorylation of YAP. Our data suggest that LRP6 has unexpected roles as a nutrient sensor and Hippo signaling regulator.

Development and Characterization of Embroidery-Based Textile Electrodes for Surface EMG Detection.

The interest in wearable devices has expanded to measurement devices for building IoT-based mobile healthcare systems and sensing bio-signal data through clothing. Surface electromyography, called sEMG, is one of the most popular bio-signals that can be applied to health monitoring systems. In general, gel-based (Ag/AgCl) electrodes are mainly used, but there are problems, such as skin irritation due to long-time wearing, deterioration of adhesion to the skin due to moisture or sweat, and low applicability to clothes. Hence, research on dry electrodes as a replacement is increasing. Accordingly, in this study, a textile-based electrode was produced with a range of electrode shapes, and areas were embroidered with conductive yarn using an embroidery technique in the clothing manufacturing process. The electrode was applied to EMG smart clothing for fitness, and the EMG signal detection performance was analyzed. The electrode shape was manufactured using the circle and wave type. The wave-type electrode was more morphologically stable than the circle-type electrode by up to 30% strain, and the electrode shape was maintained as the embroidered area increased. Skin-electrode impedance analysis confirmed that the embroidered area with conductive yarn affected the skin contact area, and the impedance decreased with increasing area. For sEMG performance analysis, the rectus femoris was selected as a target muscle, and the sEMG parameters were analyzed. The wave-type sample showed higher EMG signal strength than the circle-type. In particular, the electrode with three lines showed better performance than the fill-type electrode. These performances operated without noise, even with a commercial device. Therefore, it is expected to be applicable to the manufacture of electromyography smart clothing based on embroidered electrodes in the future.

Gastrostomy tube placement for long-term oral drug administration in non-human primates.

BACKGROUND: Non-human primates (NHPs) are often used as recipients in preclinical transplantation research that in most cases involves administration of various drugs including immunosuppressants. Long-term oral drug administration, particularly tacrolimus, is challenging in the transplant recipient NHPs. Oral drug administration method using the mixture of drug and fruit juice has been used in NHPs, but this is not always effective in all monkeys. To those monkeys who are poorly compliant, oral drug administration in restraint or administration using gastrostomy tube should be necessary. The aim of this study was to compare the efficacy of between oral drug administration in restraint and administration using gastrostomy tube and to report complications and solutions to overcome the problems related to gastrostomy tube for long-term oral drug dosing in rhesus monkeys. METHODS: Fifteen of 4- to 5-year-old male and female healthy rhesus monkeys weighing 5.0-6.8 kg were used as recipients for porcine pancreatic islet transplantation. Oral drug administration in restraint was used for four monkeys, and gastrostomy tube was placed to other 11 monkeys (8-French Feeding tube, n=6; Tri-Funnel Replacement Gastrostomy tube, n=5). Oral immunosuppressive drugs such as sirolimus and tacrolimus were administered through the tube. The efficacy and the extent of ease for administration and related complications were compared between two groups. RESULTS AND CONCLUSIONS: The complication of gastrostomy included a transient inflammation in the skin and peritonitis caused by a leakage around implantation site (one case), which could be overcome by changing suture method and tube type to interlocking box suture and Tri-Funnel Replacement Gastrostomy tube, respectively. Despite these complications, oral drug administration using gastrostomy tube allowed us to perform accurate dosage of drug administration and to reduce the stress that both the monkey and the researcher may experience. Taken together, this study showed that gastrostomy tube placement is a better alternative to oral drug administration in restraint for long-term oral drug administration in rhesus monkeys who tend to refuse to eat the mixture of drug and fruit juice.

Induction, management, and complications of streptozotocin-induced diabetes mellitus in rhesus monkeys.

BACKGROUND: Diabetes mellitus (DM) model using streptozotocin (STZ) which induces chemical ablation of ß cell in the pancreas has been widely used for various research purposes in non-human primates. However, STZ has been known to have a variety of adverse effects such as nephrotoxicity, hepatotoxicity, and even mortality. The purpose of this study is to report DM induction by STZ, toxicity associated with STZ and procedure and complication of exogenous insulin treatment for DM management in rhesus monkeys (Macaca mulatta) that are expected to be transplanted with porcine islets within 2 months. METHODS: Streptozotocin (immediately dissolved in normal saline, 110 mg/kg) was slowly infused via central catheter for 10 minutes in 22 rhesus monkeys. Clinical signs, complete blood count and blood chemistry were monitored to evaluate toxicity for 1 week after STZ injection. Monkey basal C-peptides were measured and intravenous glucose tolerance test was performed to confirm complete induction of DM. Exogenous insulin was subcutaneously injected to maintain blood glucose in diabetic rhesus monkeys and the complications were recorded while in insulin treatment. RESULTS: Severe salivation and vomiting were observed within 1 hour after STZ injection in 22 rhesus monkeys. One monkey died at 6 hours after STZ injection and the reason for the death was unknown. Pancreatitis was noticed in one monkey after STZ injection, but the monkey recovered after 5 days by medical treatment. Serum total protein and albumin decreased whereas the parameters for the liver function such as aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase significantly increased (P<.05) after STZ injection, but they were resolved within 1 week. Azotemia was not observed. Monkey fasting C-peptide levels after STZ injection were <0.1 ng/mL in 18 rhesus monkeys, but 0.34, 0.22, 0.16 ng/mL in three monkeys, respectively. The value of daily insulin requirement was 0.92±0.26IU/kg/d (range=0.45-1.29) in the monkeys. Diabetic ketoacidosis was observed in one rhesus monkeys, but the monkey recovered after 24 hours by fluid and insulin treatment. CONCLUSIONS: Streptozotocin was effective for inducing DM in rhesus monkeys, but various adverse effects such as pancreatitis, liver toxicity or death were observed. Therefore, careful and suitable medical managements should be implemented to eliminate the risks of mortality and severe adverse effects.

Fabrication of Textile-Based Dry Electrode and Analysis of Its Surface EMG Signal for Applying Smart Wear.

Ag/AgCl hydrogel electrodes, which are wet electrodes, are generally used to acquire bio-signals non-invasively. Research concerning dry electrodes is ongoing due to the following limitations of wet electrodes: (1) skin irritation and disease when attached for a long time; (2) poor adhesion due to sweat; and (3) considerable cost due to disposable use. Accordingly, electrodes in film, embroidery, and knit forms were manufactured from conductive sheets and conductive yarns, which are typical textile-type dry electrode materials, using different manufacturing methods and conditions. The prepared electrodes were conducted to measure the morphology, surface resistance, skin-electrode impedance, EMG signal acquisition, and analysis. The conductive sheet type electrode exhibited a similar skin-impedance, noise, and muscle activation signal amplitude to the Ag/AgCl gel electrode due to the excellent adhesion and shape stabilization. Embroidery electrodes were manufactured based on two-dimension lock stitch (Em_LS) and three-dimension moss-stitch (Em_MS). More stable EMG signal acquisition than Em_LS was possible when manufactured with Em_MS. The knit electrode was manufactured with the typical structures of plain, purl, and interlock. Although it was possible to acquire EMG signals, considerable noise was generated as the shape and size of the electrodes were changed due to the stretch characteristics of the knit structure. Finally, the applicability of the textile-type dry electrode was confirmed by combining it with a wearable device. More stable and accurate EMG signal acquirement will be possible through more precise parameter control in the future.

Regulation of the Low-Density Lipoprotein Receptor-Related Protein LRP6 and Its Association With Disease: Wnt/ß-Catenin Signaling and Beyond.

Wnt signaling plays crucial roles in development and tissue homeostasis, and its dysregulation leads to various diseases, notably cancer. Wnt/ß-catenin signaling is initiated when the glycoprotein Wnt binds to and forms a ternary complex with the Frizzled and low-density lipoprotein receptor-related protein 5/6 (LRP5/6). Despite being identified as a Wnt co-receptor over 20 years ago, the molecular mechanisms governing how LRP6 senses Wnt and transduces downstream signaling cascades are still being deciphered. Due to its role as one of the main Wnt signaling components, the dysregulation or mutation of LRP6 is implicated in several diseases such as cancer, neurodegeneration, metabolic syndrome and skeletal disease. Herein, we will review how LRP6 is activated by Wnt stimulation and explore the various regulatory mechanisms involved. The participation of LRP6 in other signaling pathways will also be discussed. Finally, the relationship between LRP6 dysregulation and disease will be examined in detail.

Process Optimization for Manufacturing PAN-Based Conductive Yarn with Carbon Nanomaterials through Wet Spinning.

This study aimed to manufacture PAN-based conductive yarn using a wet-spinning process. Two types of carbon nanomaterials, multiwall carbon nanotubes (MWCNT) and carbon nanofiber (CNF), were used alone or in a mixture. First, to derive the optimal composite solution condition for the wet spinning process, a composite solution was prepared with carbon nanomaterials of the same total mass weight (%) and three types of mechanical stirring were performed: mechanical stirring, ultra-sonication, and ball milling. A ball milling process was finally selected by analyzing the viscosity. Based on the above results, 8, 16, 24, and 32 wt% carbon nanomaterial/PAN composite solutions were prepared to produce wet spinning-based composite films before preparing a conductive yarn, and their physical and electrical properties were examined. By measuring the viscosity of the composite solution and the surface resistance of the composite film according to the type and content of carbon nanomaterials, a suitable range of viscosity was found from 103 cP to 105 cP, and the electrical percolation threshold was from 16 wt% carbon nanomaterial/PAN, which showed a surface resistance of 106 Ω/sq or less. Wet spinning was possible with a PAN-based composite solution with a high content of carbon nanomaterials. The crystallinity, crystal orientation, tenacity, and thermal properties were improved when CNF was added up to 24 wt%. On the other hand, the properties deteriorated when CNTs were added alone due to aggregation. Mixing CNT and CNF resulted in poorer properties than with CNF alone, but superior properties to CNT alone. In particular, the electrical properties after incorporating 8 wt% CNT/16 wt% CNF into the PAN, 106 Ω/cm was similar to the PAN-based conductive yarn containing 32 wt% CNF. Therefore, this yarn is expected to be applicable to various smart textiles and wearable devices because of its improved physical properties such as strength and conductivity.

Study on the Flame Retardancy and Hazard Evaluation of Poly(acrylonitrile-co-vinylidene chloride) Fibers by the Addition of Antimony-Based Flame Retardants.

Antimony oxide (ATO) is used mainly as a flame retardant, but it is classified as a hazardous substance. Therefore, regulations on the use of antimony trioxide (ATO(3)) and antimony pentoxide (ATO(5)) in textile products are being developed. Accordingly, there is a need for alternative flame retardants. In this study, antimony tetroxide (ATO(4)), which has higher thermal stability and resistance to acids and alkalis than ATO(3) or ATO(5), was selected to assess its use as an alternative flame retardant. First, ATO(3) or ATO(4) were added to poly(acrylonitrile-co-vinylidene chloride) (PANVDC), and the film and wet-spun fiber were prepared. The PANVDC film with flame retardants was prepared to evaluate the flame retardancy and the mechanism of action of the flame retardants. Flame retardancy analysis showed that a limiting oxygen index of 31.2% was obtained when ATO(4) was added, which was higher than when ATO(3) was used. Subsequently, PANVDC fibers with antimony oxide were manufactured and showed improved mechanical and thermal properties when ATO(4) was used, compared to when ATO(3) was tested. In addition, migration analysis due to antimony in the fiber confirmed that the elution amount was below the acceptable standard when PANVDC fibers with ATO(4) were added. Therefore, based on these results, the flame-retardant and thermal properties of antimony tetroxide were superior to antimony trioxide, and it was confirmed that ATO(4) could be used as an alternative flame retardant to ATO(3).

PARsylated transcription factor EB (TFEB) regulates the expression of a subset of Wnt target genes by forming a complex with ß-catenin-TCF/LEF1.

Wnt signaling is mainly transduced by ß-catenin via regulation of the ß-catenin destruction complex containing Axin, APC, and GSK3ß. Transcription factor EB (TFEB) is a well-known master regulator of autophagy and lysosomal biogenesis processes. TFEB's nuclear localization and transcriptional activity are also regulated by various upstream signals. In this study, we found that Wnt signaling induces the nuclear localization of TFEB and the expression of Wnt target genes is regulated by TFEB-ß-catenin-TCF/LEF1 as well as ß-catenin-TCF/LEF1 complexes. Our biochemical data revealed that TFEB is a part of the ß-catenin destruction complex, and destabilization of the destruction complex by knockdown of either Axin or APC causes nuclear localization of TFEB. Interestingly, RNA-sequencing analysis revealed that about 27% of Wnt3a-induced genes were TFEB dependent. However, these "TFEB mediated Wnt target genes" were different from TFEB target genes involved in autophagy and lysosomal biogenesis processes. Mechanistically, we found that Tankyrase (TNKS) PARsylates TFEB with Wnt ON signaling, and the nuclear localized PARsylated TFEB forms a complex with ß-catenin-TCF/LEF1 to induce the "TFEB mediated Wnt target genes". Finally, we found that in various types of cancer, the levels of TFEB mediated Wnt target genes exhibit strong correlations with the level of Axin2, which represents the activity of Wnt signaling. Overall, our data suggest that Wnt signaling induces the expression of a subset of genes that are distinct from previously known genes regulated by the ß-catenin-TCF/LEF1 complex or TFEB, by forming a transcription factor complex consisting of PARsylated TFEB and ß-catenin-TCF/LEF1.

EMG Measurement with Textile-Based Electrodes in Different Electrode Sizes and Clothing Pressures for Smart Clothing Design Optimization.

The surface electromyography (SEMG) is one of the most popular bio-signals that can be applied in health monitoring systems, fitness training, and rehabilitation devices. Commercial clothing embedded with textile electrodes has already been released onto the market, but there is insufficient information on the performance of textile SEMG electrodes because the required configuration may differ according to the electrode material. The current study analyzed the influence of electrode size and pattern reduction rate (PRR), and hence the clothing pressure (Pc) based on in vivo SEMG signal acquisition. Bipolar SEMG electrodes were made in different electrode diameters Ø 5-30 mm, and the clothing pressure ranged from 6.1 to 12.6 mmHg. The results supported the larger electrodes, and Pc showed better SEMG signal quality by showing lower baseline noise and a gradual increase in the signal to noise ratio (SNR). In particular, electrodes, Ø ≥ 20 mm, and Pc ≥ 10 mmHg showed comparable performance to Ag-Ag/Cl electrodes in current textile-based electrodes. The current study emphasizes and discusses design factors that are particularly required in the designing and manufacturing process of smart clothing with SEMG electrodes, especially as an aspect of clothing design.

Preparation and Characterization of Zinc Hydroxystannate Coated by Aluminum Phosphate and Its Application in Poly(acrylonitrile-co-vinylidene chloride).

In this study, zinc hydroxystannate ([ZnSn(OH)6], ZHS) was coated with aluminum phosphate (AlPO4, ALP) to prepare the ZHS-ALP composite. During the coating process, the reaction conditions, such as the ALP to ZHS molar ratio, were controlled, and the morphology of the products was characterized by scanning electron microscopy (SEM). The prepared composites were introduced into poly(acrylonitrile-co-vinylidene chloride) (PANVDC), and the change in compatibility between ZHS and the polymer matrix was characterized. The results showed that ALP-ZHS (1:1), which was prepared by ALP-ZHS composite molar ratio of 1:1, could improve the dispersion and compatibility of ZHS in the polymer matrix and decrease the hydrophilicity and viscosity. Moreover, the ALP-ZHS composite had a better flame-retardant effect on PANVDC than ZHS alone. PANVDC could pass the V-0 rating in UL94, particularly the highest limiting oxygen index (LOI) value of 33.2% obtained when the ALP-ZHS (1:1) composite was added to PANVDC.

Flame-Retardant Mechanism and Mechanical Properties of Wet-Spun Poly(acrylonitrile-co-vinylidene chloride) Fibers with Antimony Trioxide and Zinc Hydroxystannate.

To produce flame retardant poly(acrylonitrile-co-vinylidene chloride) (PANVDC) fibers with limiting oxygen index (LOI) values above 28%, flame retardants are added to fibers. Because antimony trioxide (ATO) used widely for PANVDC is suspected as a carcinogen, non-toxic zinc hydroxystannate (ZHS) could be the alternative for reduction of ATO usage. Moreover, a flame retardant efficiency of the combination of ATO with ZHS could be expected because it was reported that ATO resists flame in the gas phase, whereas ZHS reacts in the condensed phase. Therefore, this study discussed the flame retardant mechanisms of ATO and ZHS in PANVDC, and evaluated the efficiency of the combination. PANVDC fibers with ATO and ZHS in 15 phr were produced by wet spinning. When ZHS was added, a more cyclized structure was detected (e.g., 1-methylnaphthalene) through pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS). As a result of SEM-EDX analysis, Sb and Cl hardly remained in char layers of PANVDC-ATO; meanwhile, Zn, Sn, and Cl remained in that of PANVDC-ZHS. This implied that SbCl3 from reaction of ATO and HCl reacts in the gas phase, whereas ZnCl2 and SnCl2 from ZHS and HCl promotes the cyclization reaction of PANVDC in the condensed phase. The LOI values of PANVDC, PANVDC-ATO, and PANVDC-ZHS were 26.4%, 29.0%, and 33.5%, respectively. This suggests that ZHS is a highly effective for PANVDC. Meanwhile, the LOI of PANVDC containing ATO-ZHS mixture is 31.0%. The combination of ATO and ZHS exhibited no efficiency. The addition of ATO and ZHS slightly reduced the tenacities of the fibers, respectively, 3.11 and 3.75 from 4.42 g/den.

Preparation and Properties of Carbon Fiber/Carbon Nanotube Wet-Laid Composites.

In this study, carbon nanotubes (CNTs) were introduced into carbon fiber (CF) wet-laid composites as functional nano-fillers to fabricate multi-functional composites with improved mechanical, electrical, and thermal properties. It was considered that the wet-laid process was most suitable in order to introduce filler into brittle and rigid carbon fiber substrates, and we established the conditions of the process that could impart dispersibility and bonding between the fibers. We introduced polyamide 6 (PA6) short fiber, which is the same polymeric material as the stacking film, into carbon fiber and CNT mixture to enhance the binding interactions between carbon fiber and CNTs. Various types of CNT-reinforced carbon fiber wet-laid composites with PA6 short fibers were prepared, and the morphology, mechanical and electrical properties of the composites were estimated. As CNT was added to the carbon fiber nonwoven, the electrical conductivity increased by 500% but the tensile strength decreased slightly. By introducing short fibers of the same material as the matrix between CNT-CF wet-laid nonwovens, it was possible to find optimum conditions to increase the electrical conductivity while maintaining mechanical properties.