Interaction between the gut microbiota and colonic enteroendocrine cells regulates host metabolism.

Nutrient handling is an essential function of the gastrointestinal tract. Hormonal responses of small intestinal enteroendocrine cells (EECs) have been extensively studied but much less is known about the role of colonic EECs in metabolic regulation. To address this core question, we investigated a mouse model deficient in colonic EECs. Here we show that colonic EEC deficiency leads to hyperphagia and obesity. Furthermore, colonic EEC deficiency results in altered microbiota composition and metabolism, which we found through antibiotic treatment, germ-free rederivation and transfer to germ-free recipients, to be both necessary and sufficient for the development of obesity. Moreover, studying stool and blood metabolomes, we show that differential glutamate production by intestinal microbiota corresponds to increased appetite and that colonic glutamate administration can directly increase food intake. These observations shed light on an unanticipated host-microbiota axis in the colon, part of a larger gut-brain axis, that regulates host metabolism and body weight.

Palliative procedures for malignant gastric outlet obstruction: A network meta-analysis.

BACKGROUND: The optimal treatment for malignant gastric outlet obstruction (GOO) remains uncertain. The aim of this systematic review was to comprehensively investigate the efficacy and safety of four palliative treatments for malignant GOO: gastrojejunostomy (GJ), endoscopic ultrasound-guided gastroenterostomy (EGE), stomach-partitioning gastrojejunostomy (PGJ), and endoscopic stenting (ES). METHODS: We searched the PubMed, Embase, Cochrane Library, Scopus, and Web of Science databases; ClinicalTrials.gov; and the World Health Organization International Clinical Trials Registry Platform for randomized controlled trials (RCTs) and cohort studies comparing the aforementioned treatments for malignant GOO. We included studies that reported at least one of the following clinical outcomes: clinical success, 30-day mortality, reintervention rate, or length of hospital stay. Evidence from RCTs and non-RCTs was naïve combined to perform network meta-analysis through the frequentist approach using an inverse variance model. Treatments were ranked by P-score. RESULTS: This network meta-analysis included 3417 patients from 4 RCTs, 4 prospective cohort studies, and 32 retrospective cohort studies. PGJ was the optimal approach in terms of clinical success and reintervention (P-scores: 0.95 and 0.90, respectively). EGE had the highest probability of being the optimal treatment in terms of 30-day mortality and complications (P-scores: 0.82 and 0.99, respectively). Cluster ranking to combine the P-scores for 30-day mortality and reintervention indicated the benefits of PGJ and EGE (cophenetic correlation coefficient: 0.94; PGJ and EGE were in the same cluster). CONCLUSION: PGJ and EGE are recommended for malignant GOO. PGJ could be the alternative choice in centers with limited resources or cases unsuitable for EGE.

Prevalence and determinants of constipation in children in Asia: a systematic review and meta-analysis.

Background: Constipation is prevalent worldwide, significantly increasing healthcare costs and diminishing the quality of life in children affected. Current studies have yielded mixed results regarding the factors associated with constipation, and mainly focusing on patients outside of Asia. Moreover, most of these studies lack focus on the paediatric population. This study aimed to identify the prevalence and associated factors of constipation among children in Asia. Methods: In this systematic review and meta-analysis, we systematically searched PubMed, Scopus, and Cochrane for cohort and cross-sectional studies published from database inception up to October 12, 2022, and continued with manual searching until September 2, 2023. Eligible studies were those that included children in Asia aged 0-18 years old suffering from idiopathic constipation, with prevalence value provided in the English abstract. The analysis included clinical and general population. Children with organic constipation, who had undergone gastrointestinal surgery, or with congenital defects were excluded, as these factors affect the incidence of constipation. Data included in the analysis were extracted from published reports only. The extracted data were pooled using random-effects model to analyse the prevalence of constipation in children in Asia. This study is registered with PROSPERO, CRD42022367122. Findings: Out of 4410 systematically searched studies and 36 manually searched ones, a total of 50 studies were included in the final analysis, encompassing data from 311,660 children residing in Asia. The pooled prevalence of constipation was 12.0% (95% CI 9.3-14.6%, I2 = 99.8%). There was no significant difference in constipation prevalence observed by sex and geographical location. Nonetheless, adolescents and children aged 1-9 years exhibited a significantly higher prevalence constipation compared to infants (p < 0.0001) Additionally, significant differences in constipation rates were observed across various diagnostic methods, population sources, and mental health conditions. Interpretation: Despite the high heterogeneity resulting from varying diagnostic tools or definitions used among studies, our review adds to the literature on constipation among children in Asia. It reveals a notably high prevalence of constipation in this demographic. Diagnostic methods, age, and compromised mental health emerged as significant influencers of constipation among children in Asia, highlighting potential strategies to mitigate constipation prevalence in children in Asia. Funding: The National Science and Technology Council, Taiwan.

Strengthening cardiac therapy pipelines using human pluripotent stem cell-derived cardiomyocytes.

Advances in hiPSC isolation and reprogramming and hPSC-CM differentiation have prompted their therapeutic application and utilization for evaluating potential cardiovascular safety liabilities. In this perspective, we showcase key efforts toward the large-scale production of hiPSC-CMs, implementation of hiPSC-CMs in industry settings, and recent clinical applications of this technology. The key observations are a need for traceable gender and ethnically diverse hiPSC lines, approaches to reduce cost of scale-up, accessible clinical trial datasets, and transparent guidelines surrounding the safety and efficacy of hiPSC-based therapies.

Perlecan (HSPG2) promotes structural, contractile, and metabolic development of human cardiomyocytes.

Perlecan (HSPG2), a heparan sulfate proteoglycan similar to agrin, is key for extracellular matrix (ECM) maturation and stabilization. Although crucial for cardiac development, its role remains elusive. We show that perlecan expression increases as cardiomyocytes mature in vivo and during human pluripotent stem cell differentiation to cardiomyocytes (hPSC-CMs). Perlecan-haploinsuffient hPSCs (HSPG2+/-) differentiate efficiently, but late-stage CMs have structural, contractile, metabolic, and ECM gene dysregulation. In keeping with this, late-stage HSPG2+/- hPSC-CMs have immature features, including reduced ⍺-actinin expression and increased glycolytic metabolism and proliferation. Moreover, perlecan-haploinsuffient engineered heart tissues have reduced tissue thickness and force generation. Conversely, hPSC-CMs grown on a perlecan-peptide substrate are enlarged and display increased nucleation, typical of hypertrophic growth. Together, perlecan appears to play the opposite role of agrin, promoting cellular maturation rather than hyperplasia and proliferation. Perlecan signaling is likely mediated via its binding to the dystroglycan complex. Targeting perlecan-dependent signaling may help reverse the phenotypic switch common to heart failure.

Cytotoxicity of the field lampricide 3-trifluoromethyl-4-nitrophenol (TFM) in tadpole cell lines from North American frogs.

The common field lampricide, 3-trifluoromethyl-4-nitrophenol (TFM), is used to treat streams and creeks infested with highly invasive and destructive sea lamprey (Petromyzon marinus) in the tributaries of the Great Lakes. Unfortunately, amphibian deaths have been reported following stream treatments with TFM. Larval amphibians (tadpoles) are more susceptible to TFM toxicity than adult amphibians. The aim of this study was to test the toxicity of TFM in eight new tadpole cell lines from the green frog (Lithobates clamitans), wood frog (Lithobates sylvaticus), and American toad (Anaxyrus americanus). A cell viability bioassay using two fluorescent dyes, Alamar Blue and CFDA-AM, was performed following 24-h and 72-h exposures to a range of TFM concentrations. In general, TFM exposure reduced Alamar Blue fluorescence more rapidly than CFDA-AM fluorescence in tadpole cells, suggesting that Alamar Blue is perhaps a better diagnostic indicator of cell health for acute TFM cytotoxicity. At present, the in vivo 96-h LC50s of TFM are only available for L. clamitans and they correlated well with the in vitro EC50 values for the green frog tadpole cell lines in this study. The eight tadpole cell lines with different relative sensitivities to TFM cytotoxicity could prove to be useful tools in assessing next-generation lampricides in high-throughput bioassays to ensure safety in frogs before their sea lamprey-targeted application in the field.

Hypersensitive response to interferon-stimulated gene (ISG)-inducing double-stranded RNA in American bullfrog tadpole fibroblasts.

American bullfrogs are thought to be carriers of ranaviruses and contribute to their global spread via trade. Bullfrog tadpoles succumb to ranaviral infection's more severe and deadly effects than bullfrog adults. Presently, little is known about bullfrog tadpoles' innate antiviral immunity, possible due to the lack of available bullfrog tadpole cell lines. In this study, we describe a novel bullfrog tadpole fibroblast cell line named BullTad-leg. Its general cellular attributes, gene expression and function of class-A scavenger receptors (SR-As), and responses to poly IC (a synthetic dsRNA mimicking viral dsRNAs and a potent inducer of the interferon (IFN)-mediated antiviral responses) are investigated. Its abundant expression of vimentin corroborated with the cells' fibroblast morphology. BullTad-leg cells expressed transcripts of four SR-A members: SR-AI, SCARA3, SCARA4, and SCARA5, but transcripts of MARCO, the fifth SR-A member, were not detected. BullTad-leg cells expressed functional SR-As and could bind AcLDL. BullTad-leg cells exhibited cytotoxicity in response to poly IC treatment via SR-As. Additionally, very low doses of poly IC were able to induce dose-dependent expressions of ISGs including Mx, PKR, ISG20, and IFI35. This research sheds new light on the innate immune response, particularly SR-A biology and dsRNA responsiveness, in bullfrog tadpoles.

Chronic activation of human cardiac fibroblasts in vitro attenuates the reversibility of the myofibroblast phenotype.

Activation of cardiac fibroblasts and differentiation to myofibroblasts underlies development of pathological cardiac fibrosis, leading to arrhythmias and heart failure. Myofibroblasts are characterised by increased α-smooth muscle actin (α-SMA) fibre expression, secretion of collagens and changes in proliferation. Transforming growth factor-beta (TGF-ß) and increased mechanical stress can initiate myofibroblast activation. Reversibility of the myofibroblast phenotype has been observed in murine cells but has not been explored in human cardiac fibroblasts. In this study, chronically activated adult primary human ventricular cardiac fibroblasts and human induced pluripotent stem cell derived cFbs (hiPSC-cFbs) were used to investigate the potential for reversal of the myofibroblast phenotype using either subculture on soft substrates or TGF-ß receptor inhibition. Culture on softer plates (25 or 2 kPa Young's modulus) did not alter proliferation or reduce expression of α-SMA and collagen 1. Similarly, culture of myofibroblasts in the presence of TGF-ß inhibitor did not reverse myofibroblasts back to a quiescent phenotype. Chronically activated hiPSC-cFbs also showed attenuated response to TGF-ß receptor inhibition and inability to reverse to quiescent fibroblast phenotype. Our data demonstrate substantial loss of TGF-ß signalling plasticity as well as a loss of feedback from the surrounding mechanical environment in chronically activated human myofibroblasts.

Macrophages regulate gastrointestinal motility through complement component 1q.

Peristaltic movement of the intestine propels food down the length of the gastrointestinal tract to promote nutrient absorption. Interactions between intestinal macrophages and the enteric nervous system regulate gastrointestinal motility, yet we have an incomplete understanding of the molecular mediators of this crosstalk. Here, we identify complement component 1q (C1q) as a macrophage product that regulates gut motility. Macrophages were the predominant source of C1q in the mouse intestine and most extraintestinal tissues. Although C1q mediates the complement-mediated killing of bacteria in the bloodstream, we found that C1q was not essential for the immune defense of the intestine. Instead, C1q-expressing macrophages were located in the intestinal submucosal and myenteric plexuses where they were closely associated with enteric neurons and expressed surface markers characteristic of nerve-adjacent macrophages in other tissues. Mice with a macrophage-specific deletion of C1qa showed changes in enteric neuronal gene expression, increased neurogenic activity of peristalsis, and accelerated intestinal transit. Our findings identify C1q as a key regulator of gastrointestinal motility and provide enhanced insight into the crosstalk between macrophages and the enteric nervous system.

Investigation of variants in genetics and virulence of Porcine Epidemic Diarrhea Virus after serial passage on Vero cells.

Porcine epidemic diarrhea virus (PEDV) is a highly contagious intestinal virus. However, the current PEDV vaccine, which is produced from classical strain G1, offers low protection against recently emerged strain G2. This study aims to develop a better vaccine strain by propagating the PS6 strain, a G2b subgroup originating from Vietnam, on Vero cells until the 100th passage. As the virus was propagated, its titer increased, and its harvest time decreased. Analysis of the nucleotide and amino acid variation of the PS6 strain showed that the P100PS6 had 11, 4, and 2 amino acid variations in the 0 domain, B domain, and ORF3 protein, respectively, compared to the P7PS6 strain. Notably, the ORF3 gene was truncated due to a 16-nucleotide deletion mutation, resulting in a stop codon. The PS6 strain's virulence was evaluated in 5-day-old piglets, with P7PS6 and P100PS6 chosen for comparison. The results showed that P100PS6-inoculated piglets exhibited mild clinical symptoms and histopathological lesions, with a 100% survival rate. In contrast, P7PS6-inoculated piglets showed rapid and typical clinical symptoms of PEDV infection, and the survival rate was 0%. Additionally, the antibodies (IgG and IgA) produced from inoculated piglets with P100PS6 bound to both the P7PS6 and P100PS6 antigens. This finding suggested that the P100PS6 strain was attenuated and could be used to develop a live-attenuated vaccine against highly pathogenic and prevalent G2b-PEDV strains.

Giant Tracheal Fibroepithelial Polyp Treated Successfully by High-Frequency Electrocautery Ablation.

Endotracheal fibroepithelial polyp is a rare disease in the airways. This report describes a rare case of a tracheal giant fibroepithelial polyp. A 17-year-old woman was admitted to the hospital with severe acute respiratory failure. Chest computed tomography revealed a tumor located below the epiglottis. Endotracheal bronchoscopic examination showed a giant polyp. This endotracheal polyp was removed with ablation, by using high-frequency electricity through flexible bronchoscopy under intravenous anesthesia. The patient has had a good recovery after the intervention and at long-term follow-up. We herein describe and discuss the appropriate therapeutic approach and also review the pertinent literature.

Joint Clustering and Resource Allocation Optimization in Ultra-Dense Networks with Multiple Drones as Small Cells Using Game Theory.

In this study, we consider the combination of clustering and resource allocation based on game theory in ultra-dense networks that consist of multiple macrocells using massive multiple-input multiple-output and a vast number of randomly distributed drones serving as small-cell base stations. In particular, to mitigate the intercell interference, we propose a coalition game for clustering small cells, with the utility function being the ratio of signal to interference. Then, the optimization problem of resource allocation is divided into two subproblems: subchannel allocation and power allocation. We use the Hungarian method, which is efficient for solving binary optimization problems, to assign the subchannels to users in each cluster of small cells. Additionally, a centralized algorithm with low computational complexity and a distributed algorithm based on the Stackelberg game are provided to maximize the network energy efficiency (EE). The numerical results demonstrate that the game-based method outperforms the centralized method in terms of execution time in small cells and is better than traditional clustering in terms of EE.

Chitosan membrane drafting silver-immobilized graphene oxide nanocomposite for banana preservation: Fabrication, physicochemical properties, bioactivities, and application.

In this study, silver-immobilized graphene oxide/chitosan (AGC/CTS) membranes were assembled by the solvent evaporation method, wherein Curcuma longa extract was used to synthesize silver-immobilized graphene oxide (AGC) nanocomposite. The characterization results showed that the AGC was successfully synthesized with AgNPs distributed quite evenly on GO sheets. The as-prepared AGC also exhibited high antibacterial activity and low cytotoxicity towards normal cell lines compared to human epithelial carcinoma cell lines. Besides, the fabrication of AGC/CTS membranes was additionally assessed with different AGC ratios and thicknesses. The results revealed the membrane containing 3 wt% of AGC with great hygroscopicity and elastic modulus of 27.03 ± 3.07 MPa. The samples also performed excellent bactericidal capability, along with good mechanical properties for banana preservation. Therewithal, the membrane-coated bananas were also elucidated to ripen at slower paces and less damage, with no appearance of patches of mold on the banana peel surface, eventually prolonging the shelf life of bananas up to 10 days as compared to the non-coated ones. The aforesaid results confirm the potential application of the AGC/CTS membrane as a safe and alternative fruit preservation agent in the food industry.

X-ray-induced bio-acoustic emissions from cultured cells.

PURPOSE: We characterize for the first time the emission of acoustic waves from cultured cells irradiated with X-ray photon radiation. METHODS AND MATERIALS: Human cancer cell lines (MCF-7, HL-60) and control cell-free media were exposed to 1 Gy X-ray photons while recording the sound generated before, during and after irradiation using custom large-bandwidth ultrasound transducer. The effects of dose rate and cell viability were investigated. RESULTS: We report the first recorded acoustic signals captured from a collective pressure wave response to ionizing irradiation in cell culture. The acoustic signal was co-terminous with the radiation pulse, its magnitude was dependent on radiation dose rate, and live and dead cells showed qualitatively and quantitatively different acoustic signal characteristics. The signature of the collective acoustic peaks was temporally wider and with higher acoustic power for irradiated HL-60 than for irradiated MCF-7. CONCLUSIONS: We show that X-ray irradiation induces two cultured cancer cell types to emit a characteristic acoustic signal for the duration of the radiation pulse. The rapid decay of the signal excludes acoustic emissions themselves from contributing to the inter-organism bystander signal previously reported in intact animals, but they remain a potential component of the bystander process in tissues and cell cultures. This preliminary study suggests that further work on the potential role of radiation-induced acoustic emission (RIAE) in the inter-cellular bystander effect is merited.

Performance of vertical up-flow-constructed wetlands integrating with microbial fuel cell (VFCW-MFC) treating ammonium in domestic wastewater.

Vertical up-flow-constructed wetlands integrating with microbial fuel cell (VFCW-MFC) were evaluated for NH4+-N removal and bioelectricity recovery. The experiments were carried out in lab-scale VFCW-MFC microcosms treating synthetic domestic wastewater under different operational conditions of pH, hydraulic retention time, and mass loading rate. Effects of wild ornamental grass (Cenchrus setaceus) on treatment performance and voltage output were investigated simultaneously. Experiments demonstrated that the neutral pH of influents favoured NH4+-N removal and power generation. Extended retention time improved treatment capacity and power output but likely depended on the substrate availability. COD removal and power output increased, while NH4+-N removal decreased with increasing mass loading rates. At the loading rate of 88.31 mg COD/L.day, planted VFCW-MFCs exhibited better NH4+-N treatment performance (36.9%) and higher voltage output (132%-143%) than unplanted systems. Plants did not affect the COD removal efficiency of VFCW-MFCs (>95%). Power density was in the range of 1.26-1.59 mW/m2 in planted microcosms with a maximum CE of 13.6%. The anode layer accounted for a major proportion of NH4+-N removal in VFCW-MFCs. This study implies that NH4+-N in domestic wastewaters with relatively high COD:N ratios can be treated effectively in up-flow CW-MFCs via anaerobic processes, including anammox and heterotrophic denitrifying processes. The mass loading rate could be a critical parameter to balance different microbial processes, thus, coincidently determining the potential of power recovery from wastewaters.

Generation of a human iPSC-derived cardiomyocyte/fibroblast engineered heart tissue model.

Animal models have proven integral to broadening our understanding of complex cardiac diseases but have been hampered by significant species-dependent differences in cellular physiology. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have shown great promise in the modelling of cardiac diseases despite limitations in functional and structural maturity. 3D stem cell-derived cardiac models represent a step towards mimicking the intricate microenvironment present in the heart as an in vitro model. Incorporation of non-myocyte cell types, such as cardiac fibroblasts, into engineered heart tissue models (EHTs) can help better recapitulate the cell-to-cell and cell-to-matrix interactions present in the human myocardium. Integration of human-induced pluripotent stem cell-derived cardiac fibroblasts (hiPSC-CFs) and hiPSC-CM into EHT models enables the generation of a genetically homogeneous modelling system capable of exploring the abstruse structural and electrophysiological interplay present in cardiac pathophysiology. Furthermore, the construction of more physiologically relevant 3D cardiac models offers great potential in the replacement of animals in heart disease research. Here we describe efficient and reproducible protocols for the differentiation of hiPSC-CMs and hiPSC-CFs and their subsequent assimilation into EHTs. The resultant EHT consists of longitudinally arranged iPSC-CMs, incorporated alongside hiPSC-CFs. EHTs with both hiPSC-CMs and hiPSC-CFs exhibit slower beating frequencies and enhanced contractile force compared to those composed of hiPSC-CMs alone. The modified protocol may help better characterise the interplay between different cell types in the myocardium and their contribution to structural remodelling and cardiac fibrosis.

Disease Recognition in X-ray Images with Doctor Consultation-Inspired Model.

The application of chest X-ray imaging for early disease screening is attracting interest from the computer vision and deep learning community. To date, various deep learning models have been applied in X-ray image analysis. However, models perform inconsistently depending on the dataset. In this paper, we consider each individual model as a medical doctor. We then propose a doctor consultation-inspired method that fuses multiple models. In particular, we consider both early and late fusion mechanisms for consultation. The early fusion mechanism combines the deep learned features from multiple models, whereas the late fusion method combines the confidence scores of all individual models. Experiments on two X-ray imaging datasets demonstrate the superiority of the proposed method relative to baseline. The experimental results also show that early consultation consistently outperforms the late consultation mechanism in both benchmark datasets. In particular, the early doctor consultation-inspired model outperforms all individual models by a large margin, i.e., 3.03 and 1.86 in terms of accuracy in the UIT COVID-19 and chest X-ray datasets, respectively.

Pharmacists' Perspectives on the Use of Telepharmacy in Response to COVID-19 Pandemic in Ho Chi Minh City, Vietnam.

Introduction: Telepharmacy, the application of information and communication technologies in healthcare services, has been adopted in many countries to provide patients with pharmaceutical care. However, it has yet to be widely used in Vietnam. This study was conducted to assess the current status of use and the factors associated with the willingness to use telepharmacy of pharmacists in Vietnam. Methods: A descriptive cross-sectional study was conducted from February to July 2021; 414 pharmacists were recruited to fill in an online survey. Results: Overall, 86.7% of participants have used telepharmacy application and 87.2% of them were willing to apply telepharmacy in pharmacy practice. According to our multivariate analysis, the level of readiness was associated with positive attitude (odds ratio [OR] = 4.67; 95% confidence interval [CI]: 2.26-9.66), and a good behavior (OR = 11.34; 95% CI: 3.84-33.45). Discussion: Developing a telepharmacy system with appropriate features is essential to meet the requirements of pharmacy practice amid the spread of the COVID-19 pandemic.

Structure-based 3D-Pharmacophore modeling to discover novel interleukin 6 inhibitors: An in silico screening, molecular dynamics simulations and binding free energy calculations.

Interleukin 6 (IL-6) is a cytokine with various biological functions in immune regulation, hematopoiesis, and inflammation. Elevated IL-6 levels have been identified in several severe disorders such as sepsis, acute respiratory distress syndrome (ARDS), and most recently, COVID-19. The biological activity of IL-6 relies on interactions with its specific receptor, IL-6Rα, including the membrane-bound IL-6 receptor (mIL-6R) and the soluble IL-6 receptor (sIL-6R). Thus, inhibition of the interaction between these two proteins would be a potential treatment for IL-6 related diseases. To date, no orally available small-molecule drug has been approved. This study focuses on finding potential small molecules that can inhibit protein-protein interactions between IL-6 and its receptor IL-6Rα using its crystal structure (PDB ID: 5FUC). First, two pharmacophore models were constructed based on the interactions between key residues of IL-6 (Phe74, Phe78, Leu178, Arg179, Arg182) and IL-6Rα (Phe229, Tyr230, Glu277, Glu278, Phe279). A database of approximately 22 million compounds was screened using 3D-pharmacophore models, molecular docking models, and ADMET properties. By analyzing the interactive capability of successfully docked compounds with important amino acids, 12 potential ligands were selected for further analysis via molecular dynamics simulations. Based on the stability of the complexes, the high interactions rate of each ligand with the key residues of IL-6/IL-6Rα, and the low binding free energy calculation, two compounds ZINC83804241 and ZINC02997430, were identified as the most potential IL-6 inhibitor candidates. These results will pave the way for the design and optimization of more specific compounds to combat cytokine storm in severe coronavirus patients.