Decoding HiPSC-CM's Response to SARS-CoV-2: mapping the molecular landscape of cardiac injury.

BACKGROUND: Acute cardiac injury caused by coronavirus disease 2019 (COVID-19) increases mortality. Acute cardiac injury caused by COVID-19 requires understanding how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) directly infects cardiomyocytes. This study provides a solid foundation for related studies by using a model of SARS-CoV-2 infection in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) at the transcriptome level, highlighting the relevance of this study to related studies. SARS-CoV-2 infection in hiPSC-CMs has previously been studied by bioinformatics without presenting the full molecular biological process. We present a unique bioinformatics view of the complete molecular biological process of SARS-CoV-2 infection in hiPSC-CMs. METHODS: To validate the RNA-seq datasets, we used GSE184715 and GSE150392 for the analytical studies, GSE193722 for validation at the cellular level, and GSE169241 for validation in heart tissue samples. GeneCards and MsigDB databases were used to find genes associated with the phenotype. In addition to differential expression analysis and principal component analysis (PCA), we also performed protein-protein interaction (PPI) analysis, functional enrichment analysis, hub gene analysis, upstream transcription factor prediction, and drug prediction. RESULTS: Differentially expressed genes (DEGs) were classified into four categories: cardiomyocyte cytoskeletal protein inhibition, proto-oncogene activation and inflammation, mitochondrial dysfunction, and intracellular cytoplasmic physiological function. Each of the hub genes showed good diagnostic prediction, which was well validated in other datasets. Inhibited biological functions included cardiomyocyte cytoskeletal proteins, adenosine triphosphate (ATP) synthesis and electron transport chain (ETC), glucose metabolism, amino acid metabolism, fatty acid metabolism, pyruvate metabolism, citric acid cycle, nucleic acid metabolism, replication, transcription, translation, ubiquitination, autophagy, and cellular transport. Proto-oncogenes, inflammation, nuclear factor-kappaB (NF-κB) pathways, and interferon signaling were activated, as well as inflammatory factors. Viral infection activates multiple pathways, including the interferon pathway, proto-oncogenes and mitochondrial oxidative stress, while inhibiting cardiomyocyte backbone proteins and energy metabolism. Infection limits intracellular synthesis and metabolism, as well as the raw materials for mitochondrial energy synthesis. Mitochondrial dysfunction and energy abnormalities are ultimately caused by proto-oncogene activation and SARS-CoV-2 infection. Activation of the interferon pathway, proto-oncogene up-regulation, and mitochondrial oxidative stress cause the inflammatory response and lead to diminished cardiomyocyte contraction. Replication, transcription, translation, ubiquitination, autophagy, and cellular transport are among the functions that decline physiologically. CONCLUSION: SARS-CoV-2 infection in hiPSC-CMs is fundamentally mediated via mitochondrial dysfunction. Therapeutic interventions targeting mitochondrial dysfunction may alleviate the cardiovascular complications associated with SARS-CoV-2 infection.

The essential role of arginine biosynthetic genes in lunate conidia formation, conidiation, mycelial growth, and virulence of nematophagous fungus, Esteya vermicola CBS115803.

BACKGROUND: The pinewood nematode (Bursaphelenchus xylophilus) causes severe damage to pine trees. The nematophagous fungus, Esteya vermicola, exhibits considerable promise in the biological control of Bursaphelenchus xylophilus due to its infectivity. Notably, the lunate conidia produced by E. vermicola can infect Bursaphelenchus xylophilus. In the study, we aim to investigate the genes involved in the formation of the lunate conidia of E. vermicola CBS115803. RESULTS: Esteya vermicola CBS115803 yielded 95% lunate conidia on the complete medium (CM) and 86% bacilloid conidia on the minimal medium (MM). Transcriptomic analysis of conidia from both media revealed a significant enrichment of differentially expressed genes in the pathway related to 'cellular amino acid biosynthesis and metabolism'. Functional assessment showed that the knockout of two arginine biosynthesis genes (EV232 and EV289) resulted in defects in conidia germination, mycelial growth, lunate conidia formation, and virulence of E. vermicola CBS115803 in Bursaphelenchus xylophilus. Remarkably, the addition of arginine to the MM improved mycelial growth, conidiation and lunate conidia formation in the mutants and notably increased conidia yield and the lunate conidia ratio in the wild-type E. vermicola CBS115803. CONCLUSION: This investigation confirms the essential role of two arginine biosynthesis genes in lunate conidia formation in E. vermicola CBS115803. The findings also suggest that the supplementation of arginine to the culture medium can enhance the lunate conidia yield. These insights contribute significantly to the application of E. vermicola CBS115803 in managing Bursaphelenchus xylophilus infections. © 2023 Society of Chemical Industry.

Establishing Gene Expression and Knockout Methods in Esteya vermicola CBS115803.

Pine wilt disease, which is caused by the nematode Bursaphelenchus xylophilus, is one of the most destructive forest diseases worldwide. Esteya vermicola, a nematophagous fungus, has emerged as a promising biological control agent. However, the limited availability of gene function analysis techniques hinders further genetic modification of this fungus. In this study, we employed a combination of enzymes (driselase, snailase, and cellulase) to enzymatically degrade the cell wall of the fungus, resulting in a high yield of protoplasts. Furthermore, by utilizing 0.6 M sucrose as an osmotic pressure stabilizer, we achieved a significant protoplast regeneration rate of approximately 31%. Subsequently, we employed the polyethylene glycol-mediated protoplast transformation method to successfully establish a genetic transformation technique for E. vermicola CBS115803. Additionally, through our investigation, we identified the Olic promoter from Aspergillus nidulans, which effectively enhanced the expression of the DsRed gene encoding a red fluorescent protein in E. vermicola CBS115803. Moreover, we successfully implemented a split-marker strategy to delete the EvIPMD gene in E. vermicola CBS115803. In summary, our findings present valuable experimental methodologies for gene function analysis in E. vermicola CBS115803.

Nanodrug enhances post-ablation immunotherapy of hepatocellular carcinoma via promoting dendritic cell maturation and antigen presentation.

Thermal ablation (TA) as an effective method treating hepatocellular carcinoma (HCC) in clinics is facing great challenges of high recurrence and metastasis. Although immune-checkpoint blockade (ICB)-based immunotherapy has shown potential to inhibit recurrence and metastasis, the combination strategy of ICB and thermal ablation has shown little progress in HCC treatments. The tremendous hurdle for combining ICB with thermal ablation lies with the insufficient antigen internalization and immaturity of tumor-infiltrating dendritic cells (TIDCs) which leads to an inferior immune response to distant tumor growth and metastasis. Herein, an antigen-capturing nanoplatform, whose surface was modified with mannose as a targeting ligand, was constructed for co-delivering tumor-associated antigens (TAAs) and m6A demethylases inhibitor (i.e., fat mass and obesity associated gene (FTO) inhibitor) into TIDCs. In vivo results demonstrate that the intratumoral injection of nanodrug followed by HCC thermal ablation promotes dendritic cells (DCs) maturation, improves tumor infiltration of effector T cells and generates immune memory, which synergize with ICB treatment to inhibit the distant tumor growth and lung metastasis. Therefore, the antigen-capturing and FTO-inhibiting nanodrug holds potential to boost the ICB-based immunotherapy against HCC after thermal ablation.

Coexistence of Riehl's Melanosis, Lupus Erythematosus and Thyroiditis in a Patient.

Introduction: Riehl's melanosis (RM) is an acquired hyperpigmentation disorder, presenting diffused and reticulate brownish-gray pigmentation, preferentially on the face and neck. RM overlaps with systemic lupus erythematosus (SLE) and Hashimoto's thyroiditis has never been reported. Case: We report a case of RM patient accompanied with SLE and Hashimoto's thyroiditis of primary hypothyroidism. Progressing, diffuse, symmetric, and reticular hyperpigmentation was seen on the face, neck, and upper limbs, manifesting as typical melanosis. Skin microscopy showed diffuse black-pepper-like changes and telangiectasias. The diagnosis of SLE and primary hypothyroidism were confirmed by follow-up investigations. The hyperpigmentation turned notably lighter after 14 months of treatment with prednisone, hydroxychloroquine, and L-thyroxine. Discussion: The exact pathogenesis of RM is unclear and exposure to coal tar dyes, ultraviolet, and fragrance fixatives in cosmetics are believed to be contributing factors, while some cases involve no triggers. It is not impossible that RM is a rare skin manifestation of SLE that has never been reported. The skin hyperpigmentation in this patient was not triggered by thyroid disease. Conclusion: RM could be a skin manifestation of autoimmunity. Coexistence of RM, lupus erythematosus and thyroiditis in the same patient is rare and has never been reported.

Correction: Yang et al. A Kinesin Vdkin2 Required for Vacuole Formation, Mycelium Growth, and Penetration Structure Formation of Verticillium dahliae. J. Fungi 2022, 8, 391.

In the original publication [...].

A Kinesin Vdkin2 Required for Vacuole Formation, Mycelium Growth, and Penetration Structure Formation of Verticillium dahliae.

The soil-borne vascular fungus Verticillium dahliae infects hundreds of dicotyledonous plants, causing severe wilt diseases. During the initial colonization, V. dahliae develops a penetration peg to enable infection of cotton roots. In some phytopathogenic fungi, vacuoles play a critical role in normal formation of the infection structure. Kinesin 2 protein is associated with vacuole formation in Ustilago maydis. To identify the function of vacuoles in the V. dahliae infection structure, we identified VdKin2, an ortholog of kinesin 2, in V. dahliae and investigated its function through gene knockout. VdKin2 mutants showed severe defects in virulence and were suppressed during initial infection and root colonization based on observation of green fluorescent protein-labeled V. dahliae. We also found that deletion of VdKin2 compromised penetration peg formation and the derived septin neck. Disruption strains were viable and showed normal microsclerotia formation, whereas mycelium growth and conidial production were reduced, with shorter and more branched hyphae. Furthermore, the VdKin2 mutant, unlike wild-type V. dahliae, lacked a large basal vacuole, accompanied by a failure to generate concentrated lipid droplets. Taken together, VdKin2 regulates vacuole formation by V. dahliae, which is required for conidiation, mycelium growth, and penetration structure formation during initial plant root infection.

Identification of VdASP F2-interacting protein as a regulator of microsclerotial formation in Verticillium dahliae.

Verticillium dahliae, a notorious phytopathogenic fungus, causes vascular wilt diseases in many plant species. The melanized microsclerotia enable V. dahliae to survive for years in soil and are crucial for its disease cycle. In a previous study, we characterized the secretory protein VdASP F2 from V. dahliae and found that VdASP F2 deletion significantly affected the formation of microsclerotia under adverse environmental conditions. In this study, we clarified that VdASP F2 is localized to the cell wall. However, the underlying mechanism of VdASP F2 in microsclerotial formation remains unclear. Transmembrane ion channel protein VdTRP was identified as a candidate protein that interacts with VdASP F2 using pull-down assays followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, and interaction of VdASP F2 and VdTRP was confirmed by bimolecular fluorescence complementary and coimmunoprecipitation assays. The deletion mutant was analysed to reveal that VdTRP is required for microsclerotial production, but it is not essential for stress resistance, carbon utilization and pathogenicity of V. dahliae. RNA-seq revealed some differentially expressed genes related to melanin synthesis and microsclerotial formation were significantly downregulated in the VdTRP deletion mutants. Taken together, these results indicate that VdASP F2 regulates the formation of melanized microsclerotia by interacting with VdTRP.

Zwitterionic hydrogel for sustained release of growth factors to enhance wound healing.

Growth factors (GFs) have been well known for their therapeutic effects on wound healing. Due to their vulnerable biostability, biomaterial carriers are usually used to deliver GFs to maintain their bioactivity. Among the carriers, PEG hydrogels are the most widely applied. But the uncontrolled release of GFs and their immunogenicity dramatically retard the application of PEG hydrogels as carriers of GFs. Herein, FGF2 loaded zwitterionic sulfobetaine methacrylate (SBMA) hydrogels were developed, and it was revealed that these hydrogels were more effective in delivering FGF2 for wound healing than were PEG hydrogels. In vitro studies demonstrated that SBMA hydrogels could successfully prolong the release of FGF2, which effectively maintained the bioactivity of FGF2. Further in vivo investigation showed that SBMA hydrogels could efficiently accelerate wound regeneration by promoting granulation tissue formation, collagen deposition, cell proliferation and migration, reepithelialization and angiogenesis. All results validated that SBMA hydrogels were promising substituents of PEG hydrogels for delivering FGF2 for wound regeneration.

A novel GNAS mutation inherited from probable maternal mosaicism causes two siblings with pseudohypoparathyroidism type 1A.

Objectives Objectives Pseudohypoparathyroidism type 1A (PHP1A) is caused by maternal inheritance of GNAS mutations. It is characterized by the resistance to several hormones, primarily the parathyroid hormone (PTH), and the features of Albright's hereditary osteodystrophy. Case presentation Here, we present a family comprised two affected brothers with PHP1A and identify a novel mutation (c.277C>T) in the GNAS gene. The siblings developed a slightly different presentation in the same clinical condition. Although both patients presented with PTH resistance, which is the hallmark of PHP, the proband showed the thyroid-stimulating hormone resistance with the progression of heterotopic ossification from skin and subcutaneous tissue into deep connective tissue, while the younger brother with normocalcemia did not show the resistance to other hormones. The patients may inherit the mutation from their mother who presumably carries the mutation as a mosaicism. Conclusions Our case highlights the significance of considering mosaicism as an explanation for apparent de novo cases of pseudohypoparathyroidism.

Successful treatment of verrucous epidermal nevus with fractional micro-plasma radio-frequency technology and photodynamic therapy.

Verrucous epidermal nevus (VEN) is a skin disorder that commonly presents at birth; it is characterized by skin-colored to brown verrucous papules in a linear distribution following Blaschko's lines. Even though it is extremely rare, VEN has been associated with malignant transformation. VEN has been treated by different treatment modalities with varying and frustrating results. We introduce a new type of treatment. The fractional micro-plasma radio-frequency (RF) technology, which uses unipolar RF technology to provoke plasma spars, creating multiple controlled micro-perforations on the skin. Photodynamic therapy (PDT) is a type of technology for disease diagnosis and treatment, in which a photosensitizer gathers within the nidus and kills the diseased cells. In this report, we present a case of VEN that was successfully treated with fractional micro-plasma RF technology and PDT without side effects or complications; a follow-up was conducted after 24 months and no signs of recurrence were observed.

Hydrolysis Batteries: Generating Electrical Energy during Hydrogen Absorption.

The hydrolysis reaction of aluminum can be decoupled into a battery by pairing an Al foil with a Pd-capped yttrium dihydride (YH2 -Pd) electrode. This hydrolysis battery generates a voltage around 0.45 V and leads to hydrogen absorption into the YH2 layer. This represents a new hydrogen absorption mechanism featuring electrical energy generation during hydrogen absorption. The hydrolysis battery converts 8-15 % of the thermal energy of the hydrolysis reaction into usable electrical energy, leading to much higher energy efficiency compared to that of direct hydrolysis.

Synergism of Rare Earth Trihydrides and Graphite in Lithium Storage: Evidence of Hydrogen-Enhanced Lithiation.

The lithium storage capacity of graphite can be significantly promoted by rare earth trihydrides (REH3 , RE = Y, La, and Gd) through a synergetic mechanism. High reversible capacity of 720 mA h g-1 after 250 cycles is achieved in YH3 -graphite nanocomposite, far exceeding the total contribution from the individual components and the effect of ball milling. Comparative study on LaH3 -graphite and GdH3 -graphite composites suggests that the enhancement factor is 3.1-3.4 Li per active H in REH3 , almost independent of the RE metal, which is evident of a hydrogen-enhanced lithium storage mechanism. Theoretical calculation suggests that the active H from REH3 can enhance the Li+ binding to the graphene layer by introducing negatively charged sites, leading to energetically favorable lithiation up to a composition Li5 C16 H instead of LiC6 for conventional graphite anode.

Silica-Derived Hydrophobic Colloidal Nano-Si for Lithium-Ion Batteries.

Silica can be converted to silicon by magnesium reduction. Here, this classical reaction is renovated for more efficient preparation of silicon nanoparticles (nano-Si). By reducing the particle size of the starting materials, the reaction can be completed within 10 min by mechanical milling at ambient temperature. The obtained nano-Si with high surface reactivity are directly reacted with 1-pentanol to form an alkoxyl-functionalized hydrophobic colloid, which significantly simplifies the separation process and minimizes the loss of small Si particles. Nano-Si in 5 g scale can be obtained in one single batch with laboratory scale setups with very high yield of 89%. Utilizing the excellent dispersion in ethanol of the alkoxyl-functionalized nano-Si, surface carbon coating can be readily achieved by using ethanol soluble oligomeric phenolic resin as the precursor. The nano-Si after carbon coating exhibit excellent lithium storage performance comparable to the state of the art Si-based anode materials, featured for the high reversible capacity of 1756 mAh·g-1 after 500 cycles at a current density of 2.1 A·g-1. The preparation approach will effectively promote the development of nano-Si-based anode materials for lithium-ion batteries.

Fabrication of copper patterns on flexible substrate by patterning-adsorption-plating process.

A novel patterning-adsorption-plating process to additively fabricate copper patterns is developed. Functional ink with ion-adsorption nanoparticles was inkjet printed on PET substrate to form the patterned adsorption film. Catalytic ion was adsorbed by amino groups in the adsorption film, and catalyzed the electroless plating of copper. The mercapto groups introduced to the film enhance the reliability of the patterns. Specific solvent used in the ink increase the surface roughness of the adsorption film, leading to a better adhesion of the patterns. The prepared copper patterns show excellent conductivity about the same with bulk copper and good adhesion on PET.