Methods to Isolate Muscle Stem Cells for Cell-Based Cultured Meat Production: A Review.

Cultured meat production relies on various cell types, including muscle stem cells (MuSCs), embryonic stem cell lines, induced pluripotent cell lines, and naturally immortalized cell lines. MuSCs possess superior muscle differentiation capabilities compared to the other three cell lines, making them key for cultured meat development. Therefore, to produce cultured meat using MuSCs, they must first be effectively separated from muscles. At present, the methods used to isolate MuSCs from muscles include (1) the pre-plating method, using the ability of cells to adhere differently, which is a biological characteristic of MuSCs; (2) the density gradient centrifugation method, using the intrinsic density difference of cells, which is a physical characteristic of MuSCs; and (3) fluorescence- and magnetic-activated cell sorting methods, using the surface marker protein on the cell surface of MuSCs, which is a molecular characteristic of MuSCs. Further efficient and valuable methods for separating MuSCs are expected to be required as the cell-based cultured meat industry develops. Thus, we take a closer look at the four methods currently in use and discuss future development directions in this review.

The mechanism underlying correlation of particulate matter-induced ferroptosis with inflammasome activation and iron accumulation in macrophages.

Particulate matter (PM) is a global environmental hazard, which affects human health through free radical production, cell death induction, and immune responses. PM activates inflammasomes leading to excessive inflammatory responses and induces ferroptosis, a type of cell death. Despite ongoing research on the correlation among PM-induced ferroptosis, immune response, and inflammasomes, the underlying mechanism of this relationship has not been elucidated. In this study, we demonstrated the levels of PM-induced cell death and immune responses in murine macrophages, J774A.1 and RAW264.7, depending on the size and composition of particulate matter. PM2.5, with extraction ions, induced significant levels of cell death and immune responses; it induces lipid peroxidation, iron accumulation, and reactive oxygen species (ROS) production, which characterize ferroptosis. In addition, inflammasome-mediated cell death occurred owing to the excessive activation of inflammatory responses. PM-induced iron accumulation activates ferroptosis and inflammasome formation through ROS production; similar results were observed in vivo. These results suggest that the link between ferroptosis and inflammasome formation induced by PM, especially PM2.5 with extraction ions, is established through the iron-ROS axis. Moreover, this study can effectively facilitate the development of a new therapeutic strategy for PM-induced immune and respiratory diseases.

Low-frequency (5-Hz) stimulation of ventrolateral periaqueductal gray modulates the descending serotonergic system in the peripheral neuropathic pain.

ABSTRACT: Neuropathic pain is a type of chronic pain that entails severe prolonged sensory dysfunctions caused by a lesion of the somatosensory system. Many of those suffering from the condition do not experience significant improvement with existing medications, resulting in various side effects. In this study, Sprague-Dawley male rats were used, and long-term deep brain stimulation of the ventrolateral periaqueductal gray was conducted in a rat model of spared nerve injury. We found that 5-Hz deep brain stimulation effectively modulated mechanical allodynia and induced neuronal activation in the rostral ventromedial medulla, restoring impaired descending serotonergic system. At the spinal level, glial cells were still activated but only the 5-HT1a receptor in the spinal cord was activated, implying its inhibitory role in mechanical allodynia. This study found that peripheral neuropathy caused dysfunction in the descending serotonergic system, and prolonged stimulation of ventrolateral periaqueductal gray can modulate the pathway in an efficient manner. This work would provide new opportunities for the development of targeted and effective treatments for this debilitating disease, possibly giving us lower chances of side effects from repeated high-frequency stimulation or long-term use of medication.

Effect of Crude Polysaccharides from Ecklonia cava Hydrolysate on Cell Proliferation and Differentiation of Hanwoo Muscle Stem Cells for Cultured Meat Production.

Ecklonia cava, a brown seaweed native to the East Asian coast, is known for its unique composition, including polysaccharides, polyphenols, and phlorotannins. Fucoidan is a sulfated polysaccharide widely used as a functional ingredient in foods. This study obtained crude polysaccharides (ECC_CPS) from E. cava celluclast enzymatic hydrolysate using ethanol precipitation. ECC_CPS increased cell viability during the proliferation of Hanwoo muscle satellite cells (HMSCs). The effect of ECC_CPS on the expression of proliferation-related markers was confirmed as MYF5 and MYOD expression significantly increased, whereas PAX7 expression was maintained. The evaluation of cell migration activity has a major impact on cell proliferation and differentiation, and the cell migration index significantly increased with ECC_CPS treatment (p < 0.01). This was related to the HGF/MET pathway and FAK pathway. Treatment with ECC_CPS promoted differentiation at the cell differentiation stage, thereby increasing the expression of differentiation markers, such as MYH2, MYH7, and MYOG (p < 0.001 or p < 0.01). Therefore, our findings imply that crude polysaccharide obtained from E. cava can be an additive ingredient that enhances the proliferation and differentiation of muscle satellite cells used in the manufacture of cultured meat products.

Anti-Inflammatory Effects of Idebenone Attenuate LPS-Induced Systemic Inflammatory Diseases by Suppressing NF-κB Activation.

Inflammation is a natural protective process through which the immune system responds to injury, infection, or irritation. However, hyperinflammation or long-term inflammatory responses can cause various inflammatory diseases. Although idebenone was initially developed for the treatment of cognitive impairment and dementia, it is currently used to treat various diseases. However, its anti-inflammatory effects and regulatory functions in inflammatory diseases are yet to be elucidated. Therefore, this study aimed to investigate the anti-inflammatory effects of idebenone in cecal ligation puncture-induced sepsis and lipopolysaccharide-induced systemic inflammation. Murine models of cecal ligation puncture-induced sepsis and lipopolysaccharide-induced systemic inflammation were generated, followed by treatment with various concentrations of idebenone. Additionally, lipopolysaccharide-stimulated macrophages were treated with idebenone to elucidate its anti-inflammatory effects at the cellular level. Idebenone treatment significantly improved survival rate, protected against tissue damage, and decreased the expression of inflammatory enzymes and cytokines in mice models of sepsis and systemic inflammation. Additionally, idebenone treatment suppressed inflammatory responses in macrophages, inhibited the NF-κB signaling pathway, reduced reactive oxygen species and lipid peroxidation, and normalized the activities of antioxidant enzyme. Idebenone possesses potential therapeutic application as a novel anti-inflammatory agent in systemic inflammatory diseases and sepsis.

Application of the Nagoya Protocol to veterinary pathogens: concerns for the control of foot-and-mouth disease.

The Nagoya Protocol is an international agreement adopted in 2010 (and entered into force in 2014) which governs access to genetic resources and the fair and equitable sharing of benefits from their utilisation. The agreement aims to prevent misappropriation of genetic resources and, through benefit sharing, create incentives for the conservation and sustainable use of biological diversity. While the equitable sharing of the benefits arising from the utilisation of genetic resources is a widely accepted concept, the way in which the provisions of the Nagoya Protocol are currently being implemented through national access and benefit-sharing legislation places significant logistical challenges on the control of transboundary livestock diseases such as foot-and-mouth disease (FMD). Delays to access FMD virus isolates from the field disrupt the production of new FMD vaccines and other tailored tools for research, surveillance and outbreak control. These concerns were raised within the FMD Reference Laboratory Network and were explored at a recent multistakeholder meeting hosted by the European Commission for the Control of FMD. The aim of this paper is to promote wider awareness of the Nagoya Protocol, and to highlight its impacts on the regular exchange and utilisation of biological materials collected from clinical cases which underpin FMD research activities, and work to develop new epidemiologically relevant vaccines and other diagnostic tools to control the disease.

A Single Injection of rAAV-shmTOR in Peripheral Nerve Persistently Attenuates Nerve Injury-Induced Mechanical Allodynia.

Activation of mammalian target of rapamycin (mTOR) has been known as one of the contributing factors in nociceptive sensitization after peripheral injury. Its activation followed by the phosphorylation of downstream effectors causes hyperexcitability of primary sensory neurons in the dorsal root ganglion. We investigated whether a single injection of rAAV-shmTOR would effectively downregulate both complexes of mTOR in the long-term and glial activation as well. Male SD rats were categorized into shmTOR (n = 29), shCON (n = 23), SNI (n = 13), and Normal (n = 8) groups. Treatment groups were injected with rAAV-shmTOR or rAAV-shCON, respectively. DRG tissues and sciatic nerve were harvested for Western blot and immunohistochemical analyses. Peripheral sensitization was gradually attenuated in the shmTOR group, and it reached a peak on PID 21. Western blot analysis showed that both p-mTORC1 and p-mTORC2 were downregulated in the DRG compared to shCON and SNI groups. We also found decreased expression of phosphorylated p38 and microglial activation in the DRG. We first attempted a therapeutic strategy for neuropathic pain with a low dose of AAV injection by interfering with the mTOR signaling pathway, suggesting its potential application in pain treatment.

Liquid Crystal Polymer-Based Miniaturized Fully Implantable Deep Brain Stimulator.

A significant challenge in improving the deep brain stimulation (DBS) system is the miniaturization of the device, aiming to integrate both the stimulator and the electrode into a compact unit with a wireless charging capability to reduce invasiveness. We present a miniaturized, fully implantable, and battery-free DBS system designed for rats, using a liquid crystal polymer (LCP), a biocompatible and long-term reliable material. The system integrates the simulator circuit, the receiver coil, and a 20 mm long depth-type microelectrode array in a dome-shaped LCP package that is 13 mm in diameter and 5 mm in height. Wireless powering and control via an inductive link enable device miniaturization, allowing for full implantation and, thus, the free behavior of untethered animals. The eight-channel stimulation electrode array was microfabricated on an LCP substrate to form a multilayered system substrate, which was monolithically encapsulated by a domed LCP lid using a specialized spot-welding process. The device functionality was validated via an in vivo animal experiment using a neuropathic pain model in rats. This experiment demonstrated an increase in the mechanical withdrawal threshold of the rats with microelectrical stimulation delivered using the fully implanted device, highlighting the effectiveness of the system.

Regional brain activity of resting-state fMRI and auditory oddball ERP with multimodal approach in individuals with internet gaming disorder.

Background and aims: Resting-state brain activity may be associated with the ability to perform tasks; however, a multimodal approach involving resting-state functional magnetic resonance imaging (fMRI) and event-related potentials (ERPs) has not been widely used to investigate addictive disorders. Methods: We explored resting-state fMRI and auditory oddball ERP values from 26 with internet gaming disorder (IGD) patients and 27 age- and intelligence quotient-matched healthy controls (HCs). To assess the characteristics of resting-state fMRI, we calculated regional homogeneity (ReHo), amplitude of low-frequency fluctuation (ALFF), and fractional amplitude of low-frequency fluctuation (fALFF); we also calculated the P3 component of the ERPs. Results: Compared with HCs, the individuals with IGD exhibited significant decreases in ReHo and fALFF values in the left inferior occipital gyrus, increased ReHo and ALFF values in the right precuneus, increased ALFF in the left superior frontal gyrus, and lower P3 amplitudes in the midline centro-parietal area during the auditory ERP task. Furthermore, the regional activity of resting-state fMRI in the right inferior temporal gyrus and the occipital regions were positively correlated with the P3 amplitudes in IGD patients, whereas ReHo values of the left hippocampus and the right amygdala were negatively correlated with P3. Discussion and conclusions: Our results suggest that IGD patients have difficulty interacting effectively with cognitive function and sensory processing, although its interpretations need some cautions. The findings in this study will broaden the overall understanding of the neurobiological mechanisms that underlie IGD pathophysiology.

Simulation Method for the Physical Deformation of a Three-Dimensional Soft Body in Augmented Reality-Based External Ventricular Drainage.

OBJECTIVES: Intraoperative navigation reduces the risk of major complications and increases the likelihood of optimal surgical outcomes. This paper presents an augmented reality (AR)-based simulation technique for ventriculostomy that visualizes brain deformations caused by the movements of a surgical instrument in a three-dimensional brain model. This is achieved by utilizing a position-based dynamics (PBD) physical deformation method on a preoperative brain image. METHODS: An infrared camera-based AR surgical environment aligns the real-world space with a virtual space and tracks the surgical instruments. For a realistic representation and reduced simulation computation load, a hybrid geometric model is employed, which combines a high-resolution mesh model and a multiresolution tetrahedron model. Collision handling is executed when a collision between the brain and surgical instrument is detected. Constraints are used to preserve the properties of the soft body and ensure stable deformation. RESULTS: The experiment was conducted once in a phantom environment and once in an actual surgical environment. The tasks of inserting the surgical instrument into the ventricle using only the navigation information presented through the smart glasses and verifying the drainage of cerebrospinal fluid were evaluated. These tasks were successfully completed, as indicated by the drainage, and the deformation simulation speed averaged 18.78 fps. CONCLUSIONS: This experiment confirmed that the AR-based method for external ventricular drain surgery was beneficial to clinicians.

Black Wheat Extracts (Arriheuk) Regulate Adipogenesis and Lipolysis via Adenosine Monophosphate (AMP) Activated Protein Kinase (AMPK)/Sirtuin 1 (SIRT1) Signaling Pathways.

Polyphenols and other compounds with antioxidant properties are found in plants and are one of the main antioxidants proven to reduce body weight and the risk of insulin resistance. Still, the mechanism behind the protective effects against obesity remains unclear. Thus, the study aims to assess the impact of flavonoid-rich arriheuk extract, a purple wheat extract, on mitochondrial function using 3T3-L1 adipocytes and investigate the molecular mechanism behind its protective effects against adipogenesis and lipolysis. The study findings strongly indicate that arriheuk significantly suppressed triglyceride levels and inhibited the expression of transcription factors like C/EBPα and PPARγ in 3T3-L1 adipocytes. Furthermore, treatment with arriheuk suppressed the expression of SREBP1c and FAS proteins linked to lipogenesis. In addition, treatment with arriheuk extract decreased the mRNA levels of adipogenic transcription factors, increased glycerol release, and inhibited adipocyte differentiation. Interestingly, the arriheuk-mediated PGC-1α expression triggered mitochondrial biogenesis by promoting the AMPK phosphorylation and SIRT1 expression in adipocytes. Also, arriheuk suppressed adipogenesis and elicited browning through the AMPK- and SIRT1-associated pathways. Collectively, these findings strongly suggest that arriheuk extract regulates browning in 3T3-L1 white adipocytes by triggering the AMPK/SIRT1 pathway, indicating the prospective applications of arriheuk as a functional food to control obesity.

Near-Infrared Fluorescent Carbon Nanotube Sensors for the Plant Hormone Family Gibberellins.

Gibberellins (GAs) are a class of phytohormones, important for plant growth, and very difficult to distinguish because of their similarity in chemical structures. Herein, we develop the first nanosensors for GAs by designing and engineering polymer-wrapped single-walled carbon nanotubes (SWNTs) with unique corona phases that selectively bind to bioactive GAs, GA3 and GA4, triggering near-infrared (NIR) fluorescence intensity changes. Using a new coupled Raman/NIR fluorimeter that enables self-referencing of nanosensor NIR fluorescence with its Raman G-band, we demonstrated detection of cellular GA in Arabidopsis, lettuce, and basil roots. The nanosensors reported increased endogenous GA levels in transgenic Arabidopsis mutants that overexpress GA and in emerging lateral roots. Our approach allows rapid spatiotemporal detection of GA across species. The reversible sensor captured the decreasing GA levels in salt-treated lettuce roots, which correlated remarkably with fresh weight changes. This work demonstrates the potential for nanosensors to solve longstanding problems in plant biotechnology.

Somatosensory ECoG-based brain-machine interface with electrical stimulation on medial forebrain bundle.

Brain-machine interface (BMI) provides an alternative route for controlling an external device with one's intention. For individuals with motor-related disability, the BMI technologies can be used to replace or restore motor functions. Therefore, BMIs for movement restoration generally decode the neural activity from the motor-related brain regions. In this study, however, we designed a BMI system that uses sensory-related neural signals for BMI combined with electrical stimulation for reward. Four-channel electrocorticographic (ECoG) signals were recorded from the whisker-related somatosensory cortex of rats and converted to extract the BMI signals to control the one-dimensional movement of a dot on the screen. At the same time, we used operant conditioning with electrical stimulation on medial forebrain bundle (MFB), which provides a virtual reward to motivate the rat to move the dot towards the desired center region. The BMI task training was performed for 7 days with ECoG recording and MFB stimulation. Animals successfully learned to move the dot location to the desired position using S1BF neural activity. This study successfully demonstrated that it is feasible to utilize the neural signals from the whisker somatosensory cortex for BMI system. In addition, the MFB electrical stimulation is effective for rats to learn the behavioral task for BMI.

Specific upregulation of extracellular miR-6238 in particulate matter-induced acute lung injury and its immunomodulation.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are life-threatening diseases characterized by a severe inflammatory response and the destruction of alveolar epithelium and endothelium. ALI/ARDS is caused by pathogens and toxic environmental stimuli, such as particulate matter (PM). However, the general symptoms of ALI/ARDS are similar, and determining the cause of lung injury is often challenging. In this study, we investigated whether there is a critical miRNA that characterizes PM-induced ALI. We found that the expression of miR-6238 is specifically upregulated in lung tissue and lung-derived extracellular vesicles (EVs) in response to PM exposure. Notably, bacterial endotoxin (Lipopolysaccharide; LPS or peptidoglycan; PTG) does not induce the expression of miR-6238 in the lung. Instead, the expression of miR-155 is dramatically increased in LPS-induced ALI. We further demonstrated that human lung epithelial cells and macrophages predominantly produce miR-6238 and miR-155, respectively. Mechanistically, EV-miR-6238 is effectively internalized into alveolar macrophages (AMs) and regulates inflammatory responses in vivo. CXCL3 is a main target of miR-6238 in AMs and modulates neutrophil infiltration into the lung alveoli. Collectively, our findings suggest that miR-6238 is a novel regulator of pulmonary inflammation and a putative biomarker that distinguishes PM-induced ALI from endotoxin (LPS/PTG)-mediated ALI.

Particulate matter induces ferroptosis by accumulating iron and dysregulating the antioxidant system.

Particulate matter is an air pollutant composed of various components, and has adverse effects on the human body. Particulate matter is known to induce cell death by generating an imbalance in the antioxidant system; however, the underlying mechanism has not been elucidated. In the present study, we demonstrated the cytotoxic effects of the size and composition of particulate matter on small intestine cells. We found that particulate matter 2.5 (PM2.5) with extraction ion (EI) components (PM2.5 EI), is more cytotoxic than PM containing only polycyclic aromatic hydrocarbons (PAHs). Additionally, PM-induced cell death is characteristic of ferroptosis, and includes iron accumulation, lipid peroxidation, and reactive oxygen species (ROS) generation. Furthermore, ferroptosis inhibitor as liproxstatin-1 and iron-chelator as deferiprone attenuated cell mortality, lipid peroxidation, iron accumulation, and ROS production after PM2.5 EI treatment in human small intestinal cells. These results suggest that PM2.5 EI may increase ferroptotic-cell death by iron accumulation and ROS generation, and offer a potential therapeutic clue for inflammatory bowel diseases in human small intestinal cells. [BMB Reports 2023; 56(2): 96-101].

Analysis of suspensions and recoveries of official foot and mouth disease free status of WOAH Members between 1996 and 2020.

Foot and mouth disease was the first disease for which, in 1996, the World Organisation for Animal Health (WOAH; founded as OIE) established an official list of disease-free territories, which has helped to facilitate the trade of animals and animal products from those territories. Since that year, there have been a number of suspensions of FMD-free status which have impacted the livestock industry of the territories affected. The objective of this study is to identify factors associated with the time taken to recover FMD-free status after suspension. Historical applications submitted (between 1996 and the first semester of 2020) by WOAH Members for recognition and recovery of FMD-free status were used as the main source of data. Only FMD-free status suspensions caused by outbreaks were considered. Data on the Member's socio-economic characteristics, livestock production systems, FMD outbreak characteristics, and control strategies were targeted for the analysis. The period of time taken to recover FMD-free status was estimated using Kaplan-Meier survival curves. A Cox proportional hazard model was used to identify factors associated with the time taken to recover FMD-free status after suspension. A total of 163 territories were granted official FMD-free status during the study period. The study sample consisted of 45 FMD-free status suspensions. Africa and the Americas accounted for over 50% of FMD-free status suspensions, while over 70% of these occurred in formerly FMD-free territories where vaccination was not practiced. The study noted that implementing a stamping-out or vaccination and remove policy shortened the time to recover FMD-free status, compared with a vaccination and retain policy. Other variables associated with the outcome were the income level of the Member, Veterinary Service capacity, time taken to implement control measures, time taken until the disposal of the last FMD case, whether the territory bordered FMD-infected territories, and time elapsed since FMD freedom. This analysis will contribute toward the understanding of the main determinants affecting the time to recover the FMD free status of WOAH Members and policy processes for FMD control and elimination.

Corrigendum: Problematic Use of Alcohol and Online Gaming as Coping Strategies During the COVID-19 Pandemic: A Mini Review.

[This corrects the article DOI: 10.3389/fpsyt.2021.685964.].

Study on Volatile Profiles, Polycyclic Aromatic Hydrocarbons, and Acrylamide Formed in Welsh Onion (Allium fistulosum L.) Fried in Vegetable Oils at Different Temperatures.

Welsh onion (Allium fistulosum L.) is widely used in diverse Asian cuisines, especially in stir-fried and deep-fried foods. This study investigated the effects of different temperatures (140, 165, and 190 °C) and types of the vegetable frying oil (soybean, corn, canola, and palm oils) on the formation of volatile profiles and hazardous compounds [polycyclic aromatic hydrocarbons (PAHs) and acrylamide] in Welsh onion. Specific volatile chemical groups such as aldehydes, sulfur-containing compounds, and furans/furanones were major volatiles in Welsh onion fried (WOF). The composition of aldehydes and sulfur-containing compounds decreased, while those of furans/furanones increased when WOF samples were exposed to higher temperatures. At 190 °C, PAHs were detected at lower than the EU maximum tolerable limit (the sum of 4 PAHs, <10 µg/kg), and acrylamide was detected below 36.46 µg/kg. The integrated study of both the quality and safety properties can provide fundamental data for the industrial processing of WOF.

Effects of pH and Osmotic Changes on the Metabolic Expressions of Bacillus subtilis Strain 168 in Metabolite Pathways including Leucine Metabolism.

Bacillus subtilis is often exposed to diverse culture conditions with the aim of improving hygiene or food quality. This can lead to changes in the volatile metabolite profiles related to the quality of fermented foods. To comprehensively interpret the associated metabolic expressions, changes in intracellular primary and extracellular secondary volatile metabolites were investigated by exposing B. subtilis to an alkaline pH (BP, pH 8.0) and a high salt concentration (BS, 1 M). In particular, B. subtilis was cultured in a leucine-enriched medium to investigate the formation of leucine-derived volatile metabolites. This study observed metabolic changes in several metabolic pathways, including carbohydrate metabolism, amino acid metabolism, fatty acid metabolism, and leucine degradation. The formation of proline (an osmolyte), furans, pyrrole, and monosaccharide sugars (glucose, galactose, and fructose) was enhanced in BS, whereas fatty acid derivatives (ketones and alcohols) increased in BP. In the case of leucine degradation, 3-methyl-butanal and 3-methylbutanol could be salt-specific metabolites, while the contents of 3-methylbutanoic acid and 3-methylbutylacetate increased in BP. These results show culture condition-specific metabolic changes, especially secondary volatile metabolites related to the sensory property of foods, in B. subtilis.