Spatially organized cellular communities form the developing human heart.

The heart, which is the first organ to develop, is highly dependent on its form to function1,2. However, how diverse cardiac cell types spatially coordinate to create the complex morphological structures that are crucial for heart function remains unclear. Here we integrated single-cell RNA-sequencing with high-resolution multiplexed error-robust fluorescence in situ hybridization to resolve the identity of the cardiac cell types that develop the human heart. This approach also provided a spatial mapping of individual cells that enables illumination of their organization into cellular communities that form distinct cardiac structures. We discovered that many of these cardiac cell types further specified into subpopulations exclusive to specific communities, which support their specialization according to the cellular ecosystem and anatomical region. In particular, ventricular cardiomyocyte subpopulations displayed an unexpected complex laminar organization across the ventricular wall and formed, with other cell subpopulations, several cellular communities. Interrogating cell-cell interactions within these communities using in vivo conditional genetic mouse models and in vitro human pluripotent stem cell systems revealed multicellular signalling pathways that orchestrate the spatial organization of cardiac cell subpopulations during ventricular wall morphogenesis. These detailed findings into the cellular social interactions and specialization of cardiac cell types constructing and remodelling the human heart offer new insights into structural heart diseases and the engineering of complex multicellular tissues for human heart repair.

A Versatile Extended Stöber Approach to Monodisperse Sub-40 nm Carbon Nanospheres for Stabilizing Atomically Dispersed Fe─N4 Sites Toward Efficient Oxygen Reduction Electrocatalysis.

The development of atomically dispersed iron-nitrogen-carbon (Fe─N─C) catalysts as an alternative to precious platinum holds great potential for the substantial progress of a variety of oxygen reduction reaction (ORR)-associated energy conversion technologies. Nevertheless, the precise synthesis of Fe─N─C single atomic catalysts (SACs) with a high density of accessible active sites and pronounced electrocatalytic performance still remains an enormous challenge. Herein, an innovative extended Stöber method is designed for the controllable preparation of monodisperse small-sized N-doped carbon colloidal nanospheres (≈40 nm) anchoring atomically isolated Fe─N4 sites (abbreviated as Fe-SA@N-CNSs hereafter) with a narrow size distribution and high uniformity. Benefiting from the single Fe─N4 moieties and the unique spherical carbon substrate, the resultant Fe-SA@N-CNSs exhibit excellent ORR activity, outstanding long-term durability, and methanol tolerance in KOH electrolyte. More impressively, when further assembled into a flexible solid-state rechargeable zinc-air battery (ZAB), the Fe-SA@N-CNSs-driven ZAB delivers a higher open circuit voltage, a larger power density, and robust cycling/mechanical stability, outperforming the state-of-the-art Pt/C-based counterpart and further testifying the great potential of the as-prepared Fe-SA@N-CNSs in diverse ORR-related practical energy devices. The developed extended Stöber method provides an efficient and versatile avenue toward the preparation of a series of well-defined SACs for diverse electrocatalytic systems.

3D Printing Approaches to Engineer Cardiac Tissue.

PURPOSE OF REVIEW: Bioengineering of functional cardiac tissue composed of primary cardiomyocytes has great potential for myocardial regeneration and in vitro tissue modeling. 3D bioprinting was developed to create cardiac tissue in hydrogels that can mimic the structural, physiological, and functional features of native myocardium. Through a detailed review of the 3D printing technologies and bioink materials used in the creation of a heart tissue, this article discusses the potential of engineered heart tissues in biomedical applications. RECENT FINDINGS: In this review, we discussed the recent progress in 3D bioprinting strategies for cardiac tissue engineering, including bioink and 3D bioprinting methods as well as examples of engineered cardiac tissue such as in vitro cardiac models and vascular channels. 3D printing is a powerful tool for creating in vitro cardiac tissues that are structurally and functionally similar to real tissues. The use of human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) enables the generation of patient-specific tissues. These tissues have the potential to be used for regenerative therapies, disease modeling, and drug testing.

Crop Mapping Based on Sentinel-2 Images Using Semantic Segmentation Model of Attention Mechanism.

Using remote sensing images to identify crop plots and estimate crop planting area is an important part of agricultural remote sensing monitoring. High-resolution remote sensing images can provide rich information regarding texture, tone, shape, and spectrum of ground objects. With the advancement of sensor and information technologies, it is now possible to categorize crops with pinpoint accuracy. This study defines crop mapping as a semantic segmentation problem; therefore, a deep learning method is proposed to identify the distribution of corn and soybean using the differences in the spatial and spectral features of crops. The study area is located in the southwest of the Great Lakes in the United States, where corn and soybean cultivation is concentrated. The proposed attention mechanism deep learning model, A2SegNet, was trained and evaluated using three years of Sentinel-2 data, collected between 2019 and 2021. The experimental results show that this method is able to fully extract the spatial and spectral characteristics of crops, and its classification effect is significantly better than that of the baseline method, and it has better classification performance than other deep learning models. We cross verified the trained model on the test sets of different years through transfer learning in both spatiotemporal and spatial dimensions. Proving the effectiveness of the attention mechanism in the process of knowledge transfer, A2SegNet showed better adaptability.

Land Cover Classification of UAV Remote Sensing Based on Transformer-CNN Hybrid Architecture.

High-precision land cover maps of remote sensing images based on an intelligent extraction method are an important research field for many scholars. In recent years, deep learning represented by convolutional neural networks has been introduced into the field of land cover remote sensing mapping. In view of the problem that a convolution operation is good at extracting local features but has limitations in modeling long-distance dependence relationships, a semantic segmentation network, DE-UNet, with a dual encoder is proposed in this paper. The Swin Transformer and convolutional neural network are used to design the hybrid architecture. The Swin Transformer pays attention to multi-scale global features and learns local features through the convolutional neural network. Integrated features take into account both global and local context information. In the experiment, remote sensing images from UAVs were used to test three deep learning models including DE-UNet. DE-UNet achieved the highest classification accuracy, and the average overall accuracy was 0.28% and 4.81% higher than UNet and UNet++, respectively. It shows that the introduction of a Transformer enhances the model fitting ability.

Design and Synthesis of Stem Cell-Laden Keratin/Glycol Chitosan Methacrylate Bioinks for 3D Bioprinting.

With the advancements in tissue engineering and three-dimensional (3D) bioprinting, physiologically relevant three-dimensional structures with suitable mechanical and bioactive properties that mimic the biological tissue can be designed and fabricated. However, the available bioinks are less than demanded. In this research, the readily available biomass sources, keratin and glycol chitosan, were selected to develop a UV-curable hydrogel that is feasible for the 3D bioprinting process. Keratin methacrylate and glycol chitosan methacrylate were synthesized, and a hybrid bioink was created by combining this protein-polysaccharide cross-linked hydrogel. While human hair keratin could provide biological functions, the other composition, glycol chitosan, could further enhance the mechanical strength of the construct. The mechanical properties, degradation profile, swelling behavior, cell viability, and proliferation were investigated with various ratios of keratin methacrylate to glycol chitosan methacrylate. The composition of 2% (w/v) keratin methacrylate and 2% (w/v) chitosan methacrylate showed a significantly higher cell number and swelling percentage than other compositions and was designated as the bioink for 3D printing afterward. The feasibility of stem cell loading in the selected formula was examined with an extrusion-based bioprinter. The cells and spheroids can be successfully printed with the synthesized bioink into a specific shape and cultured. This work provides a potential option for bioinks and delivers insights into personalization research on stem cell-laden biofabricated hydrogels in the future.

Consolidative chemotherapy after definitive concurrent chemoradiotherapy for esophageal squamous cell carcinoma patients: a population based cohort study.

BACKGROUND: The role of consolidative chemotherapy (CCT) for locally advanced esophageal squamous cell carcinoma (LA-ESCC) patients treated with definitive concurrent chemoradiotherapy (dCCRT) is unclear. We aimed to compare the overall survival (OS) of those treated with vs without CCT via a population based approach. METHODS: Eligible LA-ESCC patients diagnosed between 2011 and 2017 were identified via the Taiwan Cancer Registry. We used propensity score (PS) weighting to balance observable potential confounders between groups. The hazard ratio (HR) of death and incidence of esophageal cancer mortality (IECM) were compared between those with vs without CCT. We also evaluated the OS in supplementary analyses via alternative approaches. RESULTS: Our primary analysis consisted of 368 patients in whom covariates were well balanced after PS weighting. The HR of death when CCT was compared to without was 0.67 (95% confidence interval 0.52-0.86, P = 0.002). The HR of IECM was 0.66 (P = 0.04). The HR of OS remained similarly in favor of CCT in supplementary analyses. CONCLUSIONS: We found that CCT was associated with significantly improved OS for LA-ESCC patients treated with dCCRT. Randomized controlled trials were needed to confirm this finding.

The Impact of Pretreatment PET/CT Nodal Status on Esophageal Squamous Cell Carcinoma After Neoadjuvant Chemoradiation.

BACKGROUND: For advanced esophageal cancer, the clinical significance of pretreatment nodal status (cN) as determined by different examinations remains unclear. PATIENTS AND METHODS: Patients with esophageal squamous cell carcinoma who underwent neoadjuvant chemoradiation and surgery were analyzed in this study. Pretreatment cN status assessed by CT, EUS, and PET/CT and clinicopathological features were used to evaluate tumor recurrence and long-term survival. RESULTS: Two hundred and twenty-two patients were identified in this study. Pretreatment PET/CT cN0 [odds ratio (OR) cN0 versus cN+, 5.316, p < 0.001] and pretreatment CT cN0 (OR 1.957, p = 0.032) both independently predicted ypN0. Pretreatment PET/CT cN0 was also associated with a lower recurrence rate and longer survival across the entire study group. Among patients with ypN0, pretreatment PET/CT cN+ indicated poor disease-free survival [hazard ratio (HR) 2.777, p = 0.001] and overall survival (HR 2.211, p = 0.034) compared with pretreatment PET/CT cN0, which predicted a favorable prognosis. CONCLUSIONS: Data from the current study suggest that pretreatment lymph node status as assessed by PET/CT is strongly correlated with survival outcomes after neoadjuvant chemoradiation and surgery in patients with esophageal squamous cell carcinoma. ypN0 patients can achieve better survival outcomes when pretreatment cN0 is assessed by PET/CT.

Dysphagia and masticatory performance as a mediator of the xerostomia to quality of life relation in the older population.

BACKGROUND: The impact of poor oral health on older adults' quality of life is a public health problem. In this study, the mediating effects of dental status, occlusal condition, dysphagia, and masticatory performance on the association between xerostomia and oral health-related quality of life (OHRQoL) were assessed in the older adult population. METHODS: Stratified cluster sampling was used to recruit 1076 community-dwelling adults aged 65 years and older from Kaohsiung, Taiwan. Community care centers were randomly selected according to their geographic classifications (urban, rural, or mountainous areas). Assessments of dental status and occlusal condition were performed by dentists. Information on demographics, physical function, xerostomia, dysphagia and depression was collected through face-to-face interviews. Masticatory performance was evaluated using color-changeable chewing gum. OHRQoL was measured using the Geriatric Oral Health Assessment Index. Hierarchical regression models were used to assess the relationships between OHRQoL and physical function, dental status and oral function in older adults. Path analysis was used to estimate direct and indirect pathways between xerostomia and OHRQoL. RESULTS: Participants with xerostomia exhibited a 0.20 OHRQoL reduction (p < .001) compared with patients with no xerostomia, and the direct effect accounted for 83.3% of the total effect. Dysphagia and masticatory performance were found to exert significant mediating effects on the association between xerostomia and OHRQoL (ßs = 0.20 and - 0.12, respectively; both p < .001; ßs = 0.06 and - 0.09, respectively; both p < .05). Moreover, potential mediating effects of the number of functional teeth (ßs = - 0.11 and - 0.43, respectively; both p < .001) and occlusal condition (ßs = 0.09 and 0.13, respectively; both p < .05) on the relationship between xerostomia and masticatory performance were noted. CONCLUSIONS: Dysphagia and masticatory performance may serve as pathways through which xerostomia affects quality of life. Early oral function intervention may be a valuable and actionable target for older adults to maintain quality of life. Our results further suggest that checkup and screening for oral dysfunction are essential to prevent or delay the onset of complications.

Evaluation of the necessity for chest drain placement following thoracoscopic wedge resection.

PURPOSE: To evaluate the outcomes of patients who underwent thoracoscopic wedge resection without chest drain placement. METHODS: The subjects of this retrospective study were 89 patients, who underwent thoracoscopic wedge resection at our hospital between January, 2013 and July, 2015. A total of 45 patients whose underlying condition did not meet the following criteria were assigned to the "chest drain placement group" (group A): peripheral lesions, healthy lung parenchyma, no intraoperative air leaks, hemorrhage or effusion accumulation, and no pleural adhesion. The other 44 patients whose underlying condition met the criteria were assigned to the "no chest drain placement group" (group B). Patient characteristics, specimen data, and postoperative conditions were analyzed and compared between the groups. RESULTS: Group A patients had poorer forced expiratory volume in one second (FEV1) values, less normal spirometric results, significantly higher resected lung volume, a greater maximum tumor-pleura distance, and a larger maximum tumor size. They also had a longer postoperative hospital stay. There was no difference between the two groups in postoperative complications. CONCLUSIONS: Avoiding chest drain placement after a thoracoscopic wedge resection appears to be safe and beneficial for patients who have small peripheral lesions and healthy lung parenchyma.

Shooting area of infrared camera traps affects recorded taxonomic richness and abundance of ground-dwelling invertebrates.

Ground-dwelling invertebrates are vital for soil biodiversity and function maintenance. Contemporary biodiversity assessment necessitates novel and automatic monitoring methods because of the threat of sharp reductions in soil biodiversity in farmlands worldwide. Using infrared camera traps (ICTs) is an effective method for assessing richness and abundance of ground-dwelling invertebrates. However, the influence that the shooting area of ICTs has on the diversity of ground-dwelling invertebrates has not been strongly considered during survey design. In this study, data from six ICTs with two shooting areas (A1, 38.48 cm2; A2, 400 cm2) were used to investigate ground-dwelling invertebrates in a farm in a city on the Eastern Coast of China from 20: 00 on July 31 to 00:00 on September 29, 2022. Over the course of 59 days and 1420 h, invertebrates within 9 taxa, 2447 individuals, and 112,909 ind./m2 were observed from 222,912 images. Our results show that ICTs with relatively large shooting areas recorded relatively high taxonomic richness and abundance of total ground-dwelling invertebrates, relatively high abundance of the dominant taxon, and relatively high daily and hourly abundance of most taxa. The shooting areas of ICTs significantly affected the recorded taxonomic richness and abundance of ground-dwelling invertebrates throughout the experimental period and at fine temporal resolutions. Overall, these results suggest that the shooting areas of ICTs should be considered when designing experiments, and ICTs with relatively large shooting areas are more favorable for monitoring the diversity of ground-dwelling invertebrates. This study further provides an automatic tool and high-quality data for biodiversity monitoring and protection in farmlands.

Preliminary Study on Hourly Dynamics of a Ground-Dwelling Invertebrate Community in a Farmland Vineyard.

We evaluated the hourly dynamics of ground-dwelling invertebrate communities in farmland using infrared camera traps between August and September 2022. No significant variations within 24 h nor between any two time points of each day were observed in the taxonomic richness and abundance of the entire community. However, the periods from 4:00 to 7:00 and 13:00 to 20:00 showed relatively high taxonomic richness, while those from 2:00 to 6:00 and 16:00 to 21:00 showed relatively high abundance. Millipede abundance varied significantly in a 24 h period, with higher abundance from 3:00 to 4:00 and 1:00 to 2:00. Additionally, slug, beetle, and grasshopper abundances were significantly higher from 22:00 to 23:00, 17:00 to 18:00, and 23:00 to 24:00, respectively. The abundance of other taxa did not show significant variations between any two time points of a day. Predominant generalist predators showed positive correlation in their activity times. These results suggest that significant variations within each 24 h period are uncommon at either community or taxa (except for millipedes) levels in farmland ground-dwelling invertebrates. Further, while most taxa had significantly preferred active hours, the total community did not. Therefore, hourly dynamics should be considered to understand biodiversity maintenance.

Crop classification in high-resolution remote sensing images based on multi-scale feature fusion semantic segmentation model.

The great success of deep learning in the field of computer vision provides a development opportunity for intelligent information extraction of remote sensing images. In the field of agriculture, a large number of deep convolutional neural networks have been applied to crop spatial distribution recognition. In this paper, crop mapping is defined as a semantic segmentation problem, and a multi-scale feature fusion semantic segmentation model MSSNet is proposed for crop recognition, aiming at the key problem that multi-scale neural networks can learn multiple features under different sensitivity fields to improve classification accuracy and fine-grained image classification. Firstly, the network uses multi-branch asymmetric convolution and dilated convolution. Each branch concatenates conventional convolution with convolution nuclei of different sizes with dilated convolution with different expansion coefficients. Then, the features extracted from each branch are spliced to achieve multi-scale feature fusion. Finally, a skip connection is used to combine low-level features from the shallow network with abstract features from the deep network to further enrich the semantic information. In the experiment of crop classification using Sentinel-2 remote sensing image, it was found that the method made full use of spectral and spatial characteristics of crop, achieved good recognition effect. The output crop classification mapping was better in plot segmentation and edge characterization of ground objects. This study can provide a good reference for high-precision crop mapping and field plot extraction, and at the same time, avoid excessive data acquisition and processing.

Direct and gut microbiota-mediated toxicities of environmental antibiotics to fish and aquatic invertebrates.

The accumulation of antibiotics in the environment has ecological impacts that have received less attention than the human health risks of antibiotics, although the effects could be far-reaching. This review discusses the effects of antibiotics on the health of fish and zooplankton, manifesting in direct or dysbiosis-mediated physiological impairment. Acute effects of antibiotics in these organism groups are usually induced at high concentrations (LC50 at ∼100-1000 mg/L) that are not commonly present in aquatic environments. However, when exposed to sub-lethal, environmentally relevant levels of antibiotics (ng/L-µg/L) disruption of physiological homeostasis, development, and fecundity can occur. Antibiotics at similar or lower concentrations can induce dysbiosis of gut microbiota which can affect the health of fish and invertebrates. We show that the data about molecular-level effects of antibiotics at low exposure concentrations are limited, hindering environmental risk assessment and species sensitivity analysis. Fish and crustaceans (Daphnia sp.) were the two groups of aquatic organisms used most often for antibiotic toxicity testing, including microbiota analysis. While low levels of antibiotics impact the composition and function of gut microbiota in aquatic organisms, the correlation and causality of these changes to host physiology are not straightforward. In some cases, negative or lack of correlation have occurred, and, unexpectedly, gut microbial diversity has been unaffected or increased upon exposure to environmental levels of antibiotics. Efforts to incorporate functional analyses of gut microbiota are beginning to provide valuable mechanistic information, but more data is needed for ecological risk assessment of antibiotics.

Putative adverse outcome pathways of the male reproductive toxicity derived from toxicological studies of perfluoroalkyl acids.

Adverse outcome pathway (AOP) as a conceptual framework is a powerful tool in the field of toxicology to connect seemingly discrete events at different levels of biological organizations into an organized pathway from molecular interactions to whole organism toxicity. Based on numerous toxicological studies, eight AOPs for reproductive toxicity have been endorsed by the Organization for Economic Co-operation and Development (OECD) Task Force on Hazard Assessment. We have conducted a literature survey on the mechanistic studies on male reproductive toxicity of perfluoroalkyl acids (PFAAs), a class of global environmental contaminants with high persistence, bioaccumulation and toxicity. Using the AOP development strategy, five new AOPs for male reproductive toxicity were proposed here, namely (1) changes in membrane permeability leading to reduced sperm motility, (2) disruption of mitochondrial function leading to sperm apoptosis, (3) decreased gonadotropin-releasing hormone (GnRH) expression in hypothalamus leading to reduced testosterone production in male rats, (4) activation of the p38 signaling pathway leading to disruption of BTB in mice, (5) inhibition of p-FAK-Tyr407 activity leading to the destruction of BTB. The molecular initiating events in the proposed AOPs are different from those in the endorsed AOPs, which are either receptor activation or enzyme inhibition. Although some of the AOPs are still incomplete, they can serve as a building block upon which full AOPs can be developed and applied to not only PFAAs but also other chemical toxicants with male reproductive toxicity.

Association of Salt Intake with Muscle Strength and Physical Performance in Middle-Aged to Older Chinese: The Guangzhou Biobank Cohort Study.

Older people have higher amounts of sodium accumulation in skeletal muscles than younger people, indicating the possible role of salt intake on muscular and physical function. This large population-based cross-sectional study examined the association of salt intake with muscle strength and physical performance in 4867 participants with an average age of 60.4 (standard deviation = 7.7) years. Information on salt intake was collected from self-reports. Absolute and relative grip strength (AGS and RGS), timed up-and-go test (TUGT), and falls were considered the indicators of muscle strength and physical performance. Linear and logistic regression were used to examine the associations of salt intake with AGS, RGS, TUGT score, and falls, adjusting for demographic and lifestyle factors, body mass index, self-rated health, and self-reported hypertension. Higher salt intake was independently associated with lower grip strength and a higher TUGT score. Versus light salt intake, the adjusted ß (95% confidence interval (CI)) of AGSmax, RGSmax, and TUGT scores in those with salty taste were -0.53 (-0.97, -0.08) kg, -0.04 (-0.06, -0.02) kg per kg/m2, and 0.08 (0.02, 0.14) s, respectively. A non-significant association was found between salt intake and falls. In sex-stratification analysis, the association remained in women but became non-significant in men. Our results suggest that avoiding high-salt diets may play a role in preserving muscle strength and physical function in the elderly, especially in women.

Hispidin-enriched Sanghuangporus sanghuang mycelia SS-MN4 ameliorate disuse atrophy while improving muscle endurance.

BACKGROUND: Disuse atrophy is a frequent cause of muscle atrophy, which can occur in individuals of any age who have been inactive for a prolonged period or immobilization. Additionally, acute diseases such as COVID-19 can cause frequent sequelae and exacerbate muscle wasting, leading to additional fatigue symptoms. It is necessary to investigate potent functional nutrients for muscle reinforcement in both disuse atrophy and fatigue to ensure better physical performance. METHODS: The effects of Sanghuangporus sanghuang SS-MN4 mycelia were tested on two groups of 6-week-old male mice-one with disuse atrophy and the other with fatigue. The disuse atrophy group was divided into three sub-groups: a control group, a group that underwent hind limb casting for 7 days and then recovered for 7 days and a group that was administered with SS-MN4 orally for 14 days, underwent hind limb casting for 7 days and then recovered for 7 days. The fatigue group was divided into two sub-groups: a control group that received no SS-MN4 intervention and an experimental group that was administered with SS-MN4 orally for 39 days and tested for exhaustive swimming and running on Day 31 and Day 33, respectively. RNA sequencing (RNA-seq) and western blot analysis were conducted on C2C12 cell lines to identify the therapeutic effects of SS-MN4 treatment. RESULTS: In a disuse atrophy model induced by hind limb casting, supplementing with 250 mg/kg of SS-MN4 for 14 days led to 111.2% gastrocnemius muscle mass recovery and an 89.1% improvement in motor function on a treadmill (P < 0.05). In a fatigue animal model, equivalent SS-MN4 dosage improved swimming (178.7%) and running (162.4%) activities (P < 0.05) and reduced blood urea nitrogen levels by 18% (P < 0.05). SS-MN4 treatment also increased liver and muscle glycogen storage by 34.36% and 55.6%, respectively, suggesting a higher energy reserve for exercise. RNA-seq and western blot studies from the C2C12 myotube showed that SS-MN4 extract upregulates Myh4 and helps sustain myotube integrity against dexamethasone damage. CONCLUSIONS: Supplementation of SS-MN4 (250-mg/kg body weight) with hispidin as active compound revealed a potential usage as a muscle nutritional supplement enhancing muscle recovery, fast-twitch fibre regrowth and fatigue resistance.

Phellinus linteus mycelia extract in COVID-19 prevention and identification of its key metabolic compounds profiling using UPLC-QTOF-MS/MS spectrometry.

For centuries, food, herbal medicines, and natural products have been valuable resources for discovering novel antiviral drugs, uncovering new structure-activity relationships, and developing effective strategies to prevent/treat viral infections. One such resource is Phellinus linteus, a mushroom used in folk medicine in Taiwan, Japan, Korea, and China. In this rich historical context, the key metabolites of Phellinus linteus mycelia ethanolic extract (GKPL) impacting the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at multiple stages have yet to be explored. Thus, this study systematically identifies and assesses the inhibitory effect of GKPL on the SARS-CoV-2 virus. Initially, the concentrations and contact times of GKPL against SARS-CoV-2 pseudovirus were assessed in HepG2 cells. Subsequently, utilizing the Ultra Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry method, potential biomarkers in the fungal extract were discerned. Metabolomic analysis identified 18 compounds in GKPL, with hispidin and hypholomine B present in the highest amounts. These compounds were isolated using chromatographic techniques and further identified through 1D NMR spectroscopic and mass spectrometry analysis. Hispidin and hypholomine B were found to inhibit the infection of SARS-CoV-2 pseudovirus by reducing angiotensin-converting enzyme 2 gene expression in HepG2, thereby decreasing viral entry. Moreover, hispidin and hypholomine B effectively block the spike receptor-binding domain, while hypholomine B, for the first time, showed significant inhibition of 3CL protease. This suggests that GKPL, enriched with hispidin and hypholomine B, has the potential to be used as an active ingredient against SARS-CoV-2.

High cell density and high-resolution 3D bioprinting for fabricating vascularized tissues.

Three-dimensional (3D) bioprinting techniques have emerged as the most popular methods to fabricate 3D-engineered tissues; however, there are challenges in simultaneously satisfying the requirements of high cell density (HCD), high cell viability, and fine fabrication resolution. In particular, bioprinting resolution of digital light processing-based 3D bioprinting suffers with increasing bioink cell density due to light scattering. We developed a novel approach to mitigate this scattering-induced deterioration of bioprinting resolution. The inclusion of iodixanol in the bioink enables a 10-fold reduction in light scattering and a substantial improvement in fabrication resolution for bioinks with an HCD. Fifty-micrometer fabrication resolution was achieved for a bioink with 0.1 billion per milliliter cell density. To showcase the potential application in tissue/organ 3D bioprinting, HCD thick tissues with fine vascular networks were fabricated. The tissues were viable in a perfusion culture system, with endothelialization and angiogenesis observed after 14 days of culture.

Reagent-Controlled Regiodivergent Annulations of Achmatowicz Products with Vinylogous Nucleophiles: Synthesis of Bicyclic Cyclopenta[b]pyrans and 8-Oxabicyclo[3.2.1]octane Derivatives.

Two reagent-controlled regiodivergent annulation protocols for Achmatowicz products with vinylogous nucleophiles have been developed, which furnished a series of bicyclic cyclopenta[b]pyrans and 8-oxabicyclo[3.2.1]octane derivatives in 28-90% yields. Plausible mechanisms were proposed to involve either Pd-catalyzed Tsuji-Trost allyl-allyl coupling and concomitant Michael cyclization or quinine-promoted cascade stepwise [5 + 2] cycloaddition and intramolecular Michael cyclization.