Generation of a Urine-Derived Ips Cell Line from a Patient with a Ventricular Septal Defect and Heart Failure and the Robust Differentiation of These Cells to Cardiomyocytes via Small Molecules

Background/Aims: Ventricular septal defects (VSDs) are one of the most common types of congenital heart malformations. Volume overload resulting from large VSDs can lead to heart failure (HF) and constitutes a major cause of pediatric HF with a series of often-fatal consequences. The etiology of VSD with HF is complex, and increasing evidence points toward a genetic basis. Indeed, we identified an L2483R mutation in the ryanodine receptor type 2 (RyR2) in a 2-month-old male patient with VSD with HF. Methods: We generated integration-free induced pluripotent stem cells from urine samples (UiPSCs) of this patient using Sendai virus containing the Yamanaka factors and characterized these cells based on alkaline phosphatase activity, pluripotency marker expression, and teratoma formation. Then, we induced the derived UiPSCs to rapidly and efficiently differentiate into functional cardiomyocytes through temporal modulation of canonical Wnt signaling with small molecules. Real-time PCR and immunofluorescence were used to verify the expression of myocardium-specific markers in the differentiated cardiomyocytes. The ultrastructure of the derived myocardial cells was further analyzed by using transmission electron microscopy. Results: The established UiPSC lines were positive for alkaline phosphatase activity, retained the RyR2 mutation, expressed pluripotency markers, and displayed differentiation potential to three germ layers in vivo. The UiPSC-derived cells showed hallmarks of cardiomyocytes, including spontaneous contraction and strong expression of cardiac-specific proteins and genes. However, compared with cardiomyocytes derived from H9 cells, they had a higher level of autophagy, implying that autophagy may play an important role in the development of VSD with HF. Conclusion: The protocol described here yields abundant myocardial cells and provides a solid platform for further investigation of the pathogenesis, pharmacotherapy, and gene therapy of VSD with HF

Partial recovery of respiratory function and diaphragm reinnervation following unilateral vagus nerve to phrenic nerve anastomosis in rabbits.

Respiratory dysfunction is the leading cause of mortality following upper cervical spinal cord injury (SCI). Reinnervation of the paralyzed diaphragm via an anastomosis between phrenic nerve and a donor nerve is a potential strategy to mitigate ventilatory deficits. In this study, anastomosis of vagus nerve (VN) to phrenic nerve (PN) in rabbits was performed to assess the potential capacity of the VN to compensate for lost PN inputs. At first, we compared spontaneous discharge pattern, nerve thickness and number of motor fibers between these nerves. The PN exhibited a highly rhythmic discharge while the VN exhibited a variable frequency discharge pattern. The rabbit VN had fewer motor axons (105.3±12.1 vs. 268.1±15.4). Nerve conduction and respiratory function were measured 20 weeks after left PN transection with or without left VN-PN anastomosis. Compared to rabbits subjected to unilateral phrenicotomy without VN-PN anastomosis, diaphragm muscle action potential (AP) amplitude was improved by 292%, distal latency by 695%, peak inspiratory flow (PIF) by 22.6%, peak expiratory flow (PRF) by 36.4%, and tidal volume by 21.8% in the anastomosis group. However, PIF recovery was only 28.0%, PEF 28.2%, and tidal volume 31.2% of Control. Our results suggested that VN-PN anastomosis is a promising therapeutic strategy for partial restoration of diaphragm reinnervation, but further modification and improvements are necessary to realize the full potential of this technique

Digital Twin Design

The design of Digital Twin is crucial to the Digital Roads project (and to any other projects that aim to leverage DT technologies). The DT platform is essentially a Software as a Service (SaaS). The research objectives for the Digital Twin design encapsulates: • Requirement engineering of the DT platform: eliciting user requirements, system requirements (which include functional and non-functional requirements), data requirements and more. • The design of the Domain Specific Languages (DSLs): based on the requirements, the DSLs capture the concepts (e.g. assets, attributes of assets, relationships among assets) within the DT platform. The DSLs are further broken down into: i) Foundation Data Model (FDM), a general purpose DSL to hold fundamental interoperable information for the DT; ii) Reference Data Library (RDL), an extension of FDM to model the DT for the Roads. The DSLs shall also be able to fuse information from other heterogeneous models (e.g. geometric models, building information models, imageries) for data curation. • The design of the integration cloud: the design primarily focus on how the DT product and DT process models shall be stored so that the CRUD (Create, Read, Update, Delete) operations can be performed efficiently

Flavor Chemical Profiles of Cabernet Sauvignon Wines: Six Vintages from 2013 to 2018 from the Eastern Foothills of the Ningxia Helan Mountains in China

The eastern foothills of the Helan Mountains in the Ningxia region (Ningxia), is a Chinese wine-producing region, where Cabernet Sauvignon is the main grape cultivar; however, little compositional or flavor information has been reported on Ningxia wines. Oenological parameters, volatile profiles, and phenolic profiles were determined for 98 Ningxia Cabernet Sauvignon wines from the 2013–2018 vintages, as well as 16 from Bordeaux and California, for comparison. Ningxia wines were characterized by high ethanol, low acidity, and high anthocyanin contents. Multivariate analysis revealed that citronellol and 12 characteristic phenolic compounds distinguish Ningxia wines from Bordeaux and California wines. The concentrations of most phenolic compounds were highest in the 2018 Ningxia vintage and decreased with the age of the vintage. To our knowledge, this is the first extensive regionality study on red wines from the Ningxia region

Atomistic engineering of Ag/Pt nanoclusters for remarkably boosted mass electrocatalytic activity

It is of vital importance to boost the intrinsic activity and augment the active sites of expensive and scarce platinum-based catalysts for advancing a variety of electrochemical energy applications. We herein report a mild electrochemical bottom-up approach to deposit ultrafine, but stable, Pt8Ag4 alloy clusters on carbon nanotubes (CNTs) by elaborately designing bimetallic organic cluster precursors with four silver and eight platinum atoms coordinated with µ,σ-bridged ethynylpyridine ligands, i.e., [Ag4(C24H16N4Pt)8(BF4)4]. The Pt8Ag4 cluster/CNT hybrids present impressively high platinum mass activity that is threefold that of commercial Pt/C toward the hydrogen evolution reaction, as a result of the cooperative contributions from the Ag atoms that enhance the intrinsic activity and the CNT supports that increase the activity sites. The present work affords an attractive avenue for engineering and stabilizing Pt-based nanoclusters at the atomic level and represents a promising strategy for the development of high-efficiency and durable electrocatalysts

Resource Recovery from Soybean Soaking Water by Ultrafiltration and Reverse Osmosis

Recovery of water and soluble substances from soybean soaking water (SSW) was carried out using an integrated membrane system consisting of ultrafiltration (UF) and reverse osmosis (RO). It was found that even using the four-stage UF-UF-RO-RO process with pH adjustment, the quality of RO permeate did not satisfy the emission standard, while the RO permeate from the two-stage UF-RO treatment could be reused for soybean soaking. The chemical oxygen demand (COD) of RO permeate from this process was 1700-2800 mg/L including acetic acid, gamma-aminobutyric acid, lactic acid, alcohol, and other unidentified soybean extract with -Ca center dot O or -N-H groups, which had negligible effect on the subsequent soybean soaking. The RO permeate was reused for five cycles with addition of tap water (RO permeate/tap water = 1:1.5), and the soaked soybeans were used for Koji making, which showed the similar properties with Koji made from soybeans soaked by tap water. Furthermore, the concentrates in the integrated UF-RO system provided a raw material for culturing algae or preparing animal feed. This work demonstrated that through UF-RO treatment, the wastewater pollution from food industry could be eliminated by resource recovery and utilization.</p

Records of CMAP from Control Group (A), Transection Group (B) and Bridge Group (C) rabbit showing both distal latency (dLAT) and amplitude 20 weeks after surgery.

<p>The stimulator electrode was place on the intact VN in Control Group, the proximal end of the transected VN in the Transection Group, and the VN rostral to the anastomosed site in the Bridge Group. <b>D</b>, Comparison of dLAT. <b>E</b>, Comparison of amplitude. The error bars indicate standard deviation (SD). * p<0.001 vs. control group; # p<0.001 vs. Transected Group; ## p<0.05 vs. Transected Group</p

Schematic showing the surgical preparations in the three groups.

<p><b>A.</b> Control Group: Both VN and PN were intact. <b>B.</b> Transection Group: Both VN and PN were transected. <b>C.</b> Bridge group: The proximal end of VN was anastomosed to the distal end of the PN.</p

Spontaneous efferent discharges from the PN and VN.

<p><b>A.</b> Rhythmic discharges from the PN. <b>B.</b> Sporadic, variable frequency discharges from the VN. The frequency was sometimes close to that of the PN (solid arrow). Prolonged discharges (open arrow) were only recorded from the VN. <b>C.</b> Mean discharge frequencies from the PN and VN (p = 0.021). <b>D.</b> Discharge amplitudes of PN and VN (P<0.001). The error bars indicate standard deviation (SD). * p<0.05; ** p<0.001</p