Betatrophin: a hormone that controls pancreatic ß cell proliferation.

Replenishing insulin-producing pancreatic ß cell mass will benefit both type I and type II diabetics. In adults, pancreatic ß cells are generated primarily by self-duplication. We report on a mouse model of insulin resistance that induces dramatic pancreatic ß cell proliferation and ß cell mass expansion. Using this model, we identify a hormone, betatrophin, that is primarily expressed in liver and fat. Expression of betatrophin correlates with ß cell proliferation in other mouse models of insulin resistance and during gestation. Transient expression of betatrophin in mouse liver significantly and specifically promotes pancreatic ß cell proliferation, expands ß cell mass, and improves glucose tolerance. Thus, betatrophin treatment could augment or replace insulin injections by increasing the number of endogenous insulin-producing cells in diabetics.

WUSCHEL controls genotype-dependent shoot regeneration capacity in potato.

Plant cells can reprogram their fate. The combinatorial actions of auxin and cytokinin dedifferentiate somatic cells to regenerate organs, which can develop into individual plants. As transgenic plants can be generated from genetically modified somatic cells through these processes, cell fate transition is an unavoidable step in crop genetic engineering. However, regeneration capacity closely depends on the genotype, and the molecular events underlying these variances remain elusive. In the present study, we demonstrated that WUSCHEL (WUS)-a homeodomain transcription factor-determines regeneration capacity in different potato (Solanum tuberosum) genotypes. Comparative analysis of shoot regeneration efficiency and expression of genes related to cell fate transition revealed that WUS expression coincided with regeneration rate in different potato genotypes. Moreover, in a high-efficiency genotype, WUS silencing suppressed shoot regeneration. Meanwhile, in a low-efficiency genotype, regeneration could be enhanced through the supplementation of a different type of cytokinin that promoted WUS expression. Computational modeling of cytokinin receptor-ligand interactions suggested that the docking pose of cytokinins mediated by hydrogen bonding with the core residues may be pivotal for WUS expression and shoot regeneration in potatoes. Furthermore, our whole-genome sequencing analysis revealed core sequence variations in the WUS promoters that differentiate low- and high-efficiency genotypes. The present study revealed that cytokinin responses, particularly WUS expression, determine shoot regeneration efficiency in different potato genotypes.

Abietane Diterpenoids Isolated from Torreya nucifera Disrupt Replication of Influenza Virus by Blocking the Phosphatidylinositol-3-Kinase (PI3K)-Akt and ERK Signaling Pathway.

Although vaccines and antiviral drugs are available, influenza viruses continue to pose a significant threat to vulnerable populations globally. With the emergence of drug-resistant strains, there is a growing need for novel antiviral therapeutic approaches. We found that 18-hydroxyferruginol (1) and 18-oxoferruginol (2) isolated from Torreya nucifera exhibited strong anti-influenza activity, with 50% inhibitory concentration values of 13.6 and 18.3 µM against H1N1, 12.8 and 10.8 µM against H9N2, and 29.2 µM (only compound 2) against H3N2 in the post-treatment assay, respectively. During the viral replication stages, the two compounds demonstrated stronger inhibition of viral RNA and protein in the late stages (12-18 h) than in the early stages (3-6 h). Moreover, both compounds inhibited PI3K-Akt signaling, which participates in viral replication during the later stages of infection. The ERK signaling pathway is also related to viral replication and was substantially inhibited by the two compounds. In particular, the inhibition of PI3K-Akt signaling by these compounds inhibited viral replication by sabotaging influenza ribonucleoprotein nucleus-to-cytoplasm export. These data indicate that compounds 1 and 2 could potentially reduce viral RNA and viral protein levels by inhibiting the PI3K-Akt signaling pathway. Our results suggest that abietane diterpenoids isolated from T. nucifera may be potent antiviral candidates for new influenza therapies.

Correlations with REM sleep behavior disorder severity in isolated rapid eye movement sleep behavior disorders patients.

OBJECTIVES: The pathogenesis of isolated rapid eye movement sleep behavior disorders (iRBD) is poorly understood. The severity of RBD may reflect its pathogenesis. METHODS: We compared motor function and non-motor symptoms (NMSs) between iRBD patients and healthy volunteers. We correlated motor function, NMSs, and striatal dopaminergic activity with RBD severity using video-polysomnography. RESULTS: Twenty-one iRBD patients and 17 controls participated. The Unified Parkinson's Disease Rating Scale part III scores were higher in patients compared to controls (p < 0.001). There was no difference in upper extremity function between patients and controls (right, p = 0.220; left, p = 0.209), but gait was slower in iRBD patients (walking time, p < 0.001; number of steps, p < 0.001). The mean value of the Korean version of the Mini-Mental State Exam and Clinical Dementia Rating were lower in patients (p = 0.006, p = 0.003, respectively). Patients with were also more depressed (p = 0.002), had decreased olfactory function (p < 0.001), reported more frequent sleep/fatigue episodes (p < 0.001), worse attention/memory capacity (p < 0.001), gastrointestinal problems (p = 0.009), urinary problems (p = 0.007), and pain (p = 0.083). Further, iRBD patients reported more frequent sleep-related disturbances (p = 0.004), but no difference in daytime sleepiness (p = 0.663). Disease severity was correlated with pain (r = 0.686, p = 0.002) and visuospatial function (r= -0.507, p = 0.038). There were no correlations between RBD severity and striatal dopaminergic activities (p > 0.09). CONCLUSIONS: iRBD is a multisystem neurodegenerative disorder, and gait abnormalities may be a disease characteristic, possibly related to the akinetic-rigid phenotype of Parkinson's disease. The correlation between pain/visuospatial dysfunction and RBD severity may be related to its pathogenesis.

Quantifying In Situ Structural Stabilities of Human Blood Plasma Proteins Using a Novel Iodination Protein Stability Assay.

Many of the diseases that plague society today are driven by a loss of protein quality. One method to quantify protein quality is to measure the protein folding stability (PFS). Here, we present a novel mass spectrometry (MS)-based approach for PFS measurement, iodination protein stability assay (IPSA). IPSA quantifies the PFS by tracking the surface-accessibility differences of tyrosine, histidine, methionine, and cysteine under denaturing conditions. Relative to current methods, IPSA increases protein coverage and granularity to track the PFS changes of a protein along its sequence. To our knowledge, this study is the first time the PFS of human serum proteins has been measured in the context of the blood serum (in situ). We show that IPSA can quantify the PFS differences between different transferrin iron-binding states in near in vivo conditions. We also show that the direction of the denaturation curve reflects the in vivo surface accessibility of the amino acid residue and reproducibly reports a residue-specific PFS. Along with IPSA, we introduce an analysis tool Chalf that provides a simple workflow to calculate the residue-specific PFS. The introduction of IPSA increases the potential to use protein structural stability as a structural quality metric in understanding the etiology and progression of human disease. Data is openly available at Chorusproject.org (project ID 1771).

Phosphodiesterase 5 Inhibitor Potentiates Epigallocatechin 3-O-Gallate-Induced Apoptotic Cell Death via Activation of the cGMP Signaling Pathway in Caco-2 Cells.

Epigallocatechin 3-O-gallate (EGCG) is a predominant component in green tea with various health benefits. The 67 kDa laminin receptor (67LR) is a nonintegrin cell surface receptor that is overexpressed in various types of cancer; 67LR was identified a cell surface EGCG target that plays a pivotal role in tumor growth, metastasis, and resistance to chemotherapy. However, the plasma concentration of EGCG is limited, and its molecular mechanisms remain unelucidated in colon cancer. In this study, we found that the phosphodiesterase 5 (PDE5) inhibitor, vardenafil (VDN), potentiates EGCG-induced apoptotic cell death in colon cancer cells. The combination of EGCG and VDN induced apoptosis via activation of the endothelial nitric oxide synthase/cyclic guanosine monophosphate/protein kinase Cδ signaling pathway. In conclusion, the PDE5 inhibitor, VDN, may reduce the intracellular PDE5 enzyme activity that potentiates EGCG-induced apoptotic cell death in Caco-2 cells. These results suggest that PDE5 inhibitors can be used to elevate cGMP levels to induce 67LR-mediated, cancer-specific cell death. Therefore, EGCG may be employed as a therapeutic candidate for colon cancer.

Nonviral gene editing via CRISPR/Cas9 delivery by membrane-disruptive and endosomolytic helical polypeptide.

Effective and safe delivery of the CRISPR/Cas9 gene-editing elements remains a challenge. Here we report the development of PEGylated nanoparticles (named P-HNPs) based on the cationic α-helical polypeptide poly(γ-4-((2-(piperidin-1-yl)ethyl)aminomethyl)benzyl-l-glutamate) for the delivery of Cas9 expression plasmid and sgRNA to various cell types and gene-editing scenarios. The cell-penetrating α-helical polypeptide enhanced cellular uptake and promoted escape of pCas9 and/or sgRNA from the endosome and transport into the nucleus. The colloidally stable P-HNPs achieved a Cas9 transfection efficiency up to 60% and sgRNA uptake efficiency of 67.4%, representing an improvement over existing polycation-based gene delivery systems. After performing single or multiplex gene editing with an efficiency up to 47.3% in vitro, we demonstrated that P-HNPs delivering Cas9 plasmid/sgRNA targeting the polo-like kinase 1 (Plk1) gene achieved 35% gene deletion in HeLa tumor tissue to reduce the Plk1 protein level by 66.7%, thereby suppressing the tumor growth by >71% and prolonging the animal survival rate to 60% within 60 days. Capable of delivering Cas9 plasmids to various cell types to achieve multiplex gene knock-out, gene knock-in, and gene activation in vitro and in vivo, the P-HNP system offers a versatile gene-editing platform for biological research and therapeutic applications.

Streptomyces fodineus sp. nov., an actinobacterium with antifungal activity isolated from mine area soil.

A novel actinobacterial strain producing an antifungal substance was isolated from a sample of acidic mine area soil, and its taxonomic position was evaluated. The novel strain, designated TW1S1T, formed white-grey aerial mycelium and yellow substrate mycelium on oatmeal agar. Growth occurred at 10-45 °C (optimum, 30 °C), pH 4-9 (pH 6-7) and in the presence of up to 8 % (w/v) NaCl. Melanin was produced on peptone-yeast extract-iron agar. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that the novel strain should be assigned to the genus Streptomyces, and the closest species was Streptomyces puniciscabiei S77T with 99.1 % sequence similarity, which was followed by Streptomyces durhamensis NRRL B-3309T (99.0 %), Streptomyces filipinensis NBRC 12860T (98.9 %) and Streptomyces yaanensis Z4T (98.7 %). The chemotaxonomic properties were consistent with those of Streptomyces. ll-Diaminopimelic acid was the diagnostic diamino acid, and alanine, glutamic acid and glycine were present in the peptidoglycan. The cell-wall hydrolysate also contained galactose, glucose, mannose and ribose. The predominant isoprenoid quinones were MK-9(H4) and MK-9(H6), the major polar lipids were phosphatidylglycerol and an unidentified phospholipid, and the main fatty acids were iso-C16 : 0 and anteiso-C15 : 0. However, strain TW1S1T could be distinguished from its neighbouring species by its phenotypic properties. In addition, the genome-based comparison with the closest species indicated that strain TW1S1T should be recognized as a separate species. The phylogenetic, phenotypic and chemotaxonomic as well as genomic evidence supported that TW1S1T represents a novel species of Streptomyces, for which the name Streptomycesfodineus sp. nov. is proposed (type strain, TW1S1T = KCTC 49013T = JCM 32404T).

Comparison of Prevalence and Ultrasonography Features of Thyroglossal Duct Cyst in Adults According to Radioactive Iodine Ablation.

BACKGROUND This study aimed to evaluate the prevalence of thyroglossal duct cysts (TGDCs) on ultrasonography (US) and US features of TGDCs in adults, and to assess whether the prevalence or size of TGDCs increases after radioactive iodine ablation (RIA). MATERIAL AND METHODS Between July and December 2018, 2820 patients underwent thyroid or neck US examination, performed by 2 radiologists, at our center. On the basis of real-time US, the presence or absence of TGDCs was prospectively investigated by 2 radiologists. Among the 2820 patients, 54 patients who were <19 years of age or had a radiation therapy history to the neck were excluded. Eventually, 2766 patients were included. RESULTS Of the 2766 patients, 160 (5.8%) showed a TGDC on US. The mean size of TGDCs in RIA history (+) (n=36) and RIA history (-) (n=124) groups was 0.92±0.41 cm and 0.86±0.45 cm, respectively. There was no significant difference in size of TGDCs between RIA history (+) and RIA history (-) groups (p=0.684). Between the TGDC (+) and TGDC (-) groups, there was no significant difference in patient age, gender, reason for thyroid/neck US, type of thyroid surgery, and session number and application/no application of RIA (p>0.05). The prevalence rate of TGDCs in radiologist A and B was 4.9% (70/1427) and 6.7% (90/1339), respectively. TGDCs were more common in the suprahyoid neck, and the common shapes of TGDCs were flat-to-ovoid and round. CONCLUSIONS RIA may not be associated with the prevalence or enlargement of TGDCs.

Utility of Preoperative Ultrasonography in Transferred Patients with Suspicious Malignancy on Ultrasonography-Guided Fine-Needle Aspiration Cytology of Thyroid Nodules: A Single-Center Retrospective Study.

BACKGROUND This study aimed to assess the utility and characteristics of preoperative ultrasonography (US) in patients transferred to referral hospitals from local clinics with a diagnosis of malignancy on US-guided fine-needle aspiration cytology of thyroid nodules. MATERIAL AND METHODS From January 2018 to June 2018, 109 transferred patients underwent preoperative US in our hospital for suspected thyroid malignancy on cytological analysis after US-guided fine-needle aspiration of thyroid nodules in local clinics. Preoperative US was performed by a single radiologist in all patients. Among them, 6 were excluded from the study because of refusal of thyroid surgery. Preoperative US and histopathological results were compared in all patients. RESULTS After thyroid surgery, pathological examination revealed papillary thyroid carcinoma (PTC) (n=98), follicular adenoma (n=1), and nodular hyperplasia (n=4). Of the 103 patients, 91 exhibited suspicious US findings on the preoperative US, whereas 12 did not. In the 91 patients with suspicious US findings, PTC (n=90) and follicular adenoma (n=1) were confirmed after thyroid surgery. In the 12 patients with no suspicious US findings, PTC (n=8) and nodular hyperplasia (n=4) were confirmed after thyroid surgery. On repeat analysis of the cytological slides of the 4 nodular hyperplasia cases from the local clinics, Bethesda category II (n=1) and III (n=3) were determined. CONCLUSIONS In the transferred patients with a malignant cytology, preoperative US might be helpful to detect false-positive cytology cases.

Flexible transparent conducting films with embedded silver networks composed of bimodal-sized nanoparticles for heater application.

A facile one-pot synthetic method for preparing the Ag nanoparticle inks with a bimodal size distribution was newly devised and they were successfully employed as a conducting filler to form the metal-mesh type transparent conducting electrodes on the flexible substrate. Bimodal-sized Ag nanoparticles were synthesized through the polyol process, and their size variation was occurred via finely tuned composition ratio between Ag+ ions and polymeric capping agents. The prepared bimodal-sized Ag nanoparticles exhibited the form of well-dispersed Ag nanoparticle inks without adding any dispersants and dispersion process. By filling the patterned micro-channels engraved on the flexible polymer substrate using a bimodal-sized Ag nanoparticle ink, a metal-mesh type transparent electrode (transmittance: 90% at 550 nm, haze: 1.5, area: 8 × 8 cm2) was fabricated. By applying DC voltage to the mesh type electrode, a flexible transparent joule heater was successfully achieved with a performance of 4.5 °C s-1 heat-up rate at a low input power density.

Smart Contract-Based Review System for an IoT Data Marketplace.

Internet of Things (IoT)-based devices, especially those used for home automation, consist of their own sensors and generate many logs during a process. Enterprises producing IoT devices convert these log data into more useful data through secondary processing; thus, they require data from the device users. Recently, a platform for data sharing has been developed because the demand for IoT data increases. Several IoT data marketplaces are based on peer-to-peer (P2P) networks, and in this type of marketplace, it is difficult for an enterprise to trust a data owner or the data they want to trade. Therefore, in this study, we propose a review system that can confirm the reputation of a data owner or the data traded in the P2P data marketplace. The traditional server-client review systems have many drawbacks, such as security vulnerability or server administrator's malicious behavior. However, the review system developed in this study is based on Ethereum smart contracts; thus, this system is running on the P2P network and is more flexible for the network problem. Moreover, the integrity and immutability of the registered reviews are assured because of the blockchain public ledger. In addition, a certain amount of gas is essential for all functions to be processed by Ethereum transactions. Accordingly, we tested and analyzed the performance of our proposed model in terms of gas required.

Engineering mesenchymal stem cells for regenerative medicine and drug delivery.

Researchers have applied mesenchymal stem cells (MSC) to a variety of therapeutic scenarios by harnessing their multipotent, regenerative, and immunosuppressive properties with tropisms toward inflamed, hypoxic, and cancerous sites. Although MSC-based therapies have been shown to be safe and effective to a certain degree, the efficacy remains low in most cases when MSC are applied alone. To enhance their therapeutic efficacy, researchers have equipped MSC with targeted delivery functions using genetic engineering, therapeutic agent incorporation, and cell surface modification. MSC can be genetically modified virally or non-virally to overexpress therapeutic proteins that complement their innate properties. MSC can also be primed with non-peptidic drugs or magnetic nanoparticles for enhanced efficacy and externally regulated targeting, respectively. Furthermore, MSC can be functionalized with targeting moieties to augment their homing toward therapeutic sites using enzymatic modification, chemical conjugation, or non-covalent interactions. These engineering techniques are still works in progress, requiring optimization to improve the therapeutic efficacy and targeting effectiveness while minimizing any loss of MSC function. In this review, we will highlight the advanced techniques of engineering MSC, describe their promise and the challenges of translation into clinical settings, and suggest future perspectives on realizing their full potential for MSC-based therapy.

Relationships between transporter expression and the status of BRAF V600E mutation and F-18 FDG uptake in papillary thyroid carcinomas.

PURPOSE: The purpose of this study was to evaluate the expression of the glucose transporters GLUT1 and GLUT3 in papillary thyroid carcinomas (PTCs) and to elucidate their relationship with the BRAF V600E mutation and F-18 FDG uptake. MATERIALS AND METHODS: We retrospectively analyzed data of 52 PTC patients (41 women and 11 men; mean age, 52.4 ± 14.5 years). F-18 FDG PET/CT was performed preoperatively, and the maximum standardized uptake value (SUVmax) was calculated. GLUT1/GLUT3 expression was determined immunohistochemically, and the BRAF V600E mutation was detected using DNA sequencing. RESULTS: GLUT1 and GLUT3 were expressed in 82.7% (43/52) and 59.6% (31/52) PTCs, respectively. The BRAF V600E mutation was detected in 65.4% (34/52) PTCs. The odds ratio between GLUT1 expression and the BRAF V600E mutation was 5.2 (95% CI, 1.11-24.05; p < 0.05), and that between GLUT3 expression and the BRAF V600E mutation was 3.8 (95% CI, 1.14-12.53; p < 0.05). The SUVmax of PTCs was significantly higher if they carried the BRAF V600E mutation (11.3 ± 2.0, compared with 5.7 ± 1.4 for wild type BRAF tumors, Mann-Whitney test, p = 0.016). Neither GLUT1 nor GLUT3 expression was significantly associated with the SUVmax of F-18 FDG PET/CT in PTCs. CONCLUSIONS: Our findings confirmed that both GLUT1 and GLUT3 are strongly expressed by PTCs, although their expression was not significantly associated with the SUVmax of F-18 FDG PET/CT. However, GLUT1 and GLUT3 expressions were significantly associated with the presence of the BRAF V600E mutation, and the SUVmax of tumors was significantly higher in the presence of the mutated BRAF gene.

Generation of G gene-deleted viral hemorrhagic septicemia virus (VHSV) and evaluation of its vaccine potential in olive flounder (Paralichthys olivaceus).

Generation of recombinant viruses lacking an essential gene for the production of infective viral particles would be a way to produce safety-enhanced live viral vaccines. The rhabdoviral envelope-spiked glycoprotein (G) plays critical roles in the attachment of viruses on the cell surface receptor and in the transfer of endocytosed viruses to the cytoplasm by fusion to the endosomal membrane. In the present study, we produced a G gene-lacking recombinant viral hemorrhagic septicemia virus (rVHSV-ΔG) that has no ability to form plaques in the cells without a trans-supply of the G protein, which suggests that rVHSV-ΔG is a single cycle virus and progenies of rVHSV-ΔG are not infectious. One of the major advantages of attenuated vaccines is the maintenance of replication ability in the host, which enforces the adaptive immune responses. However, in spite of lacking an ability to produce infective viral particles, immunization with the present rVHSV-ΔG induced significantly higher serum neutralization titers and survival rates against virulent VHSV challenge in olive flounder (Paralichthys olivaceus) fingerlings, indicating that the present rVHSV-ΔG has a high potential as a prophylactic vaccine.

Effect of truncation of the N-terminal region of the viral hemorrhagic septicemia virus (VHSV) P protein on viral replication.

The phosphoprotein (P) of viral hemorrhagic septicemia virus (VHSV) plays an essential role in viral replication by interconnecting the L protein and the N protein-RNA complex. In this study, to investigate the role of the N-terminal region of the P protein in viral replication, we mutated the first or the first and second or the first, second, and third ATG codon into TGA stop codons. The respective mutants were named P1, P2, and P3. Recombinant VHSVs containing each mutated P gene (rVHSV-P1, -P2, and -P3) were successfully generated by supplying the intact P protein in trans. The rVHSV-P2 and -P3 were not generated from cells expressing truncated P proteins (P1, P2 or P3 protein), but the rVHSV-P1 produced infectious viruses, even in cells without any P-protein-expressing plasmids. Nucleotide sequence analysis of the P gene of rVHSV-P1 showed that a mutation had occurred that resulted in the fourth amino acid (isoleucine, ATT) being changed to methionine (ATG) without a frameshift (P0.5), suggesting that strong selection pressure might facilitate mutations that are advantageous or essential for virus replication. Infectious rVHSV-P2 and -P3 were produced in cells expressing the P0.5 protein, suggesting that the first three amino acids of the P protein of VHSV are dispensable for viral replication. Furthermore, although the P1 protein was shorter than the P0.5 protein by only two amino acid residues, no viruses were produced when the P1 protein was supplied indicating that the fourth and the fifth amino acid residues are indispensable for normal P protein functions involved in viral replication.

Characterization and preparation of bio-tubular scaffolds for fabricating artificial vascular grafts by combining electrospinning and a 3D printing system.

The last decade has seen artificial blood vessels composed of natural polymer nanofibers grafted into human bodies to facilitate the recovery of damaged blood vessels. However, electrospun nanofibers (ENs) of biocompatible materials such as chitosan (CTS) suffer from poor mechanical properties. This study describes the design and fabrication of artificial blood vessels composed of a blend of CTS and PCL ENs and coated with PCL strands using rapid prototyping technology. The resulting tubular vessels exhibited excellent mechanical properties and showed that this process may be useful for vascular reconstruction.