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Despite their significance in receptor-mediated internalization and continued signal transduction in cells, early/sorting endosomes (EE/SE) remain incompletely characterized, with many outstanding questions that surround the dynamics of their size and number. While several studies have reported increases in EE/SE size and number resulting from endocytic events, few studies have addressed such dynamics in a methodological and quantitative manner. Herein we apply quantitative fluorescence microscopy to measure the size and number of EE/SE upon internalization of two different ligands: transferrin and epidermal growth factor. Additionally, we used siRNA knock-down to determine the involvement of 5 different endosomal RAB proteins (RAB4, RAB5, RAB8A, RAB10 and RAB11A) in EE/SE dynamics. Our study provides new information on the dynamics of endosomes during endocytosis, an important reference for researchers studying receptor-mediated internalization and endocytic events.

Therapeutic monoclonal antibodies (mAb) targeting the immune checkpoint inhibitor programmed cell death protein 1 (PD-1) have achieved considerable clinical success in anti-cancer therapy through relieving T cell exhaustion. Blockade of PD-1 interaction with its ligands PD-L1 and PD-L2 is an important determinant in promoting the functional recovery of exhausted T cells. Here, we show that anti-PD-1 mAbs act through an alternative mechanism leading to the downregulation of PD-1 surface expression on memory CD4+ and CD8+ T cells. PD-1 receptor downregulation is a distinct process from receptor endocytosis and occurs in a CD14+ monocyte dependent manner with the CD64/Fcγ receptor I acting as the primary factor for this T cell extrinsic process. Importantly, downregulation of surface PD-1 strongly enhances antigen-specific functional recovery of exhausted PD-1+CD8+ T cells. Our study demonstrates a novel mechanism for reducing cell surface levels of PD-1 and limiting the inhibitory targeting by PD-L1/2 and thereby enhancing the efficacy of anti-PD-1 Ab in restoring T cell functionality. [ABSTRACT FROM AUTHOR]

The growing interest in graphene derivatives is a result of their variety of applications in many fields. Due to their use, the oral route could be a potential way of entrance for the general population. This work assesses the biotransformation of reduced graphene oxide (rGO) after an in vitro digestion procedure (mouth, gastric, intestinal, and colon digestion), and its toxic effects in different cell models (HepG2, Caco-2, and 3D intestinal model). The characterization of rGO digestas evidenced the agglomeration of samples during the in vitro gastrointestinal (g.i.) digestion. Internalization of rGO was only evident in Caco-2 cells exposed to the colonic phase and no cellular defects were observed. Digestas of rGO did not produce remarkable cytotoxicity in any of the experimental models employed at the tested concentrations (up to 200 µg/mL), neither an inflammatory response. Undigested rGO has shown cytotoxic effects in Caco-2 cells, therefore these results suggest that the digestion process could prevent the systemic toxic effects of rGO. However, additional studies are necessary to clarify the interaction of rGO with the g.i. tract and its biocompatibility profile. [ABSTRACT FROM AUTHOR]

A value is an outward or inward view of what is "worthwhile" and is a belief system that people uses to distinguish between good and bad and to guide their behavior. Values vary from culture to culture and from time to time. In order to develop a vocabulary for measuring the values of contemporary Chinese, an open-ended survey was conducted in Study 1 to collect the value vocabulary of the Chinese public, and a text analysis was conducted in Study 2 to acquire the value vocabulary of the Chinese society. In Study 3, a word list of contemporary Chinese values was developed by integrating the words obtained from the above two studies and combining words from previous studies, and a preliminary application of the word list was conducted. The results revealed that Chinese values include four dimensions: self-fulfillment, self-cultivation, social development, and interpersonal ethics. Chinese values are characterized by diversity, and some of the socially advocated values have been internalized into the value system of individuals. [ABSTRACT FROM AUTHOR]

The consumption of fresh produce is increasing due to its role in promoting a healthy and balanced diet. However, this trend is accompanied by increased foodborne disease cases associated with pathogens such as Escherichia, Listeria, and Salmonella. Previous studies provided evidence that the internalization of foodborne pathogens in fresh produce may be a potential contamination route and may pose a public health risk. This study investigates the combination effects of storage temperature and humidity on Salmonella internalization in six types of leafy greens (iceberg lettuce, romaine lettuce, red lettuce, green onion, spinach, and kale) during the storage stage. The results indicated that temperature plays a critical role in Salmonella internalization, with higher concentrations observed in samples stored at 25 °C compared to those stored at 7 °C. The mean concentration of internalized Salmonella in the iceberg lettuce sample was the highest and that in the green onion sample was the lowest (iceberg lettuce > red lettuce > romaine lettuce > spinach > kale > green onion). Mist conditions also had an impact on internalization. The group treated with mist showed an increase in Salmonella internalization of about 10–30% rather than the group without mist treatment. This research emphasizes the importance of understanding the factors influencing bacterial internalization in fresh produce and highlights the need for proper storage conditions to minimize the risk of contamination and ensure food safety. [ABSTRACT FROM AUTHOR]

In the central nervous system (CNS), cannabinoid receptor 1 (CB1R) is preferentially expressed in axons where it has a unique property, namely resistance to agonist-driven endocytosis. This review aims to summarize what we know about molecular mechanisms of CB1R cell surface stability in axonal compartments, how these impact CB1R signaling, and to consider their physiological consequences. This review then focuses on a potential candidate for maintaining axonal CB1R at the cell surface, Src homology 3-domain growth factor receptor-bound 2-like endophilin interacting protein 1 (SGIP1). SGIP1 may contribute to the polarized distribution of CB1R and modify its signaling in axons. In addition, deletion of SGIP1 results in discrete behavioral changes in modalities controlled by the endocannabinoid system in vivo. Several drugs acting directly via CB1R have important therapeutic potential, however their adverse effects limit their clinical use. Future studies might reveal chemical approaches to target the SGIP1-CB1R interaction, with the aim to exploit the endocannabinoid system pharmaceutically in a discrete way, with minimized undesired consequences. [ABSTRACT FROM AUTHOR]

Lipid nano-systems were prepared and characterized in a series of well-established in vitro tests that could assess their interactions with the hCMEC/D3 and SH-SY5Y cell lines as a model for the blood–brain barrier and neuronal function, accordingly. The prepared formulations of nanoliposomes and nanostructured lipid carriers were characterized by z-average diameters of ~120 nm and ~105 nm, respectively, following a unimodal particle size distribution (PDI < 0.3) and negative Z-potential (−24.30 mV to −31.20 mV). Stability studies implied that the nano-systems were stable in a physiologically relevant medium as well as human plasma, except nanoliposomes containing poloxamer on their surface, where there was an increase in particle size of ~26%. The presence of stealth polymer tends to decrease the amount of adsorbed proteins onto a particle's surface, according to protein adsorption studies. Both formulations of nanoliposomes were characterized by a low cytotoxicity, while their cell viability was reduced when incubated with the highest concentration (100 μg/mL) of nanostructured lipid formulations, which could have been associated with the consumption of cellular energy, thus resulting in a reduction in metabolic active cells. The uptake of all the nano-systems in the hCMEC/D3 and SH-SY5Y cell lines was successful, most likely following ATP-dependent internalization, as well as transport via passive diffusion. [ABSTRACT FROM AUTHOR]

Objectives Negative age stereotypes have negative, assimilative effects on the subjective aging experience due to internalization processes, but sometimes positive contrast effects are reported as well, reflecting dissociation and downward comparisons. Our aim was thus to compare short-term and long-term consequences of age stereotypes on the subjective aging experience, to test the hypothesis that contrast effects are visible cross-sectionally, whereas internalization processes are observed when considering long-term changes. Methods We assessed age stereotypes and subjective age in a core sample of N = 459 participants (initial age range 30–80 years) from the Ageing as Future project across 3 consecutive measurement occasions spanning a longitudinal interval of 10 years. Short-term and long-term effects were estimated with latent growth models by assessing the effects of age stereotypes on the intercepts (cross-sectional) and on the slopes (longitudinal) of subjective age, respectively, while controlling for current self-views. Results Age stereotypes had opposite effects on subjective age depending on the time frame. A cross-sectional contrast effect was found, whereas longitudinal effects were assimilative in nature. Discussion Our findings support the time-dependent nature of the effects of age stereotypes on the subjective aging experience. Negative age stereotypes temporarily lead to a significantly younger subjective age, indicating dissociation from one's age group and downward comparison. In the long run, however, negative (positive) age stereotypes become internalized into the self-views of older people and are linked to a relatively older (younger) subjective age. [ABSTRACT FROM AUTHOR]

Copyright of Journal of International Business Studies is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In recent years, rural tourism promoting has experienced unusual development opportunities due to the positive effects of live streaming. Despite the previous literature emphasizing the importance of rural tourism live streams, the current study investigates the interaction effect between different types of broadcasters and rural tours on potential tourists' travel intentions. Through a controlled experimental design with 194 participants, this study found that the congruence between broadcasters and rural tours significantly influences potential tourists' travel intentions. Specifically, when villager (vs. celebrity) broadcasters recommend local culture tours in live streaming, potential tourists' travel intentions are significantly higher, and this effect is mediated by internalization. On the other hand, when celebrity (vs. villager) broadcasters recommend natural ecology tours, potential tourists' travel intentions are significantly higher, and this effect is caused by identification. This study contributes to the literature on rural tourism and provides practical implications for tourism marketers and broadcasters seeking to maximize the effectiveness of their promotion in live streaming. [ABSTRACT FROM AUTHOR]

Since the era of Piaget and Vygotsky, private speech (PS) has been widely discussed, but in recent years, the avenues for its study have greatly expanded. In this study, we explored the use of a recoding scheme for PS inspired by the studies of Pyotr Galperin. A coding scheme for social speech, PS, and the lack of speech, as the form of action (FA) has been proposed (i.e., external social speech, external audible speech, inaudible speech, and mental FA when no speech was produced). An exploratory study was conducted to elucidate the appropriateness of the coding scheme, both ontogenetically and during tasks. The results showed that both the coding scheme by type of speech and FA were adequate for differentiating ontogenetically between children. However, only the coding schemes of the FA were appropriate for differentiating between children as a function of their performance (i.e., time and scores) in a Tower of London task. Moreover, Galperin's scheme was more suitable when there was redundancy in performance between those with audible and inaudible external speech. [ABSTRACT FROM AUTHOR]

The interplay between membrane subregions and receptor tyrosine kinases (RTK) will influence signaling in both normal and pathological RTK conditions. In this study, epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptor β (PDGFR-β) internalizations were investigated by immunofluorescent microscopy following simultaneous treatment with EGF and PDGF-BB. We found that the two receptors utilize separate routes of internalization, which merges in a common perinuclear endosomal compartment after 45 min of stimulation. This is further strengthened when contrasting the recruitment of either EGFR or PDGFR-β to either clathrin or caveolin-1: PDGFR-β dissociates from caveolin-1 upon stimulation, and engages clathrin, whilst an increased recruitment of EGFR, to both clathrin and caveolin-1, was observed upon EGF stimulation. The association between EGFR and caveolin-1 is supported by the observation that EGFR was localized in lipid raft associated fractions, whereas PDGFR-β was not. We also found that disruption of lipid rafts using MβCD led to an increased EGFR dimerization and phosphorylation in response to ligand, as well as a dramatic decrease in AKT- and a smaller but robust decrease in ERK1/2 phosphorylation. This suggest that lipid rafts may be important to effectively connect the EGFR with downstream proteins to facilitate signaling. Our data implies that cholesterol depletion of the plasma membrane affect the signaling of EGFR and PDGFRβ differently. • EGFR phosphorylation and dimerization increases after cholesterol removal. • EGF-dependent activation of downstream AKT and ERK1/2 is reduced upon cholesterol removal. • EGFR and PDGFR-β may internalize separately but later coalesce in perinuclear vesicles. • EGFR and PDGFR-β deviate in their clathrin-independent internalization. [ABSTRACT FROM AUTHOR]

Background and Purpose: Glucagon‐like peptide‐2 (GLP‐2) is secreted postprandially by enteroendocrine L‐cells and stimulates growth of the gut and bone. One GLP‐2 analogue is approved for short bowel syndrome (SBS). To improve therapeutic efficacy, we developed biased GLP‐2 receptor (GLP‐2R) agonists through N‐terminal modifications. Experimental Approach: Variants with Ala and Trp substitutions of the first seven positions of GLP‐2(1‐33) were studied in vitro for affinity, G protein activation (cAMP accumulation), recruitment of β‐arrestin 1 and 2, and internalization of the human and mouse GLP‐2R. The intestinotrophic actions of the most efficacious (cAMP) biased variant were examined in mice. Key Results: Ala substitutions had more profound effects than Trp substitutions. For both, alterations at positions 1, 3 and 6 most severely impaired activity. β‐arrestin recruitment was more affected than cAMP accumulation. Among Ala substitutions, [H1A], [D3A] and [F6A] impaired potency (EC50) for cAMP‐accumulation >20‐fold and efficacy (Emax) to 48%–87%, and were unable to recruit arrestins. The Trp substitutions, [A2W], [D3W] and [G4W] were partial agonists (Emax of 46%–59%) with 1.7–12‐fold decreased potencies in cAMP and diminished β‐arrestin recruitment. The biased variants, [F6A], [F6W] and [S7W] induced less GLP‐2R internalization compared with GLP‐2, which induced internalization in a partly arrestin‐independent manner. In mice, [S7W] enhanced gut trophic actions with increased weight of the small intestine, increased villus height and crypt depth compared with GLP‐2. Conclusion and Implications: G protein‐biased GLP‐2R agonists with diminished receptor desensitization have superior intestinotrophic effects and may represent improved treatment of intestinal insufficiency including SBS. [ABSTRACT FROM AUTHOR]

The article examines the peculiarities of internalization of negative external effects of industrial production. The expediency of using the vertical model of internalization in relation to the negative external effects of industrial production is substantiated. The shortcomings of the mechanism of redistribution of internalization funds within the framework of the vertical model of internalization of negative external effects of industrial production applied in Ukraine are identified. It was established that the environmental tax in Ukraine does not cover public losses caused by the negative impact of pollution on the environment. A combined approach to the redistribution of internalization funds has been developed, which provides financing of environmental protection measures at the cost of the collected from the sale of permits for the emission of pollutants and parallel subsidization of the recipients of negative external effects from the fund, which is replenished from the payment of the environmental tax. It is expected that the use of a combined approach will contribute to ensuring an acceptable level of negative impact of industrial pollution on the environment, while at the same time protecting the interests of the most vulnerable sectors of the economy. It has been proven that the use of a combined approach to the redistribution of internalization funds will not have a negative impact on the development of industry in Ukraine. [ABSTRACT FROM AUTHOR]

GPR15 is a G protein‐coupled receptor involved in immune disorders such as human immunodeficiency virus‐induced enteropathy, multiple sclerosis, and colitis. Yet, the important endocytosis mechanism of GPR15 remained unclear. This study determined the participation of endocytic machinery proteins, including Gα proteins, G protein‐coupled receptor kinases (GRKs), protein kinase C, arrestins, clathrin, caveolin, and dynamin in GPR15 internalization. The results demonstrate that GPR15 internalization is moderately dependent on GRKs and clathrin, and highly dependent on caveolin and dynamin. Moreover, a bystander arrestin recruitment assay showed that GPR15 recruits arrestin‐3 to the cell membrane upon agonist stimulation, although GPR15 internalizes in an arrestin‐independent manner. Overall, our study provides novel insights into β‐arrestin recruitment and receptor internalization mechanisms for the recently deorphanized GPR15. [ABSTRACT FROM AUTHOR]

Recent research has examined predictors of false confessions in criminal investigations. By incorporating a commonly utilized social rejection paradigm (Cyberball), the current study adds to the list of personality and situational predictors by examining the role of social exclusion—which is a likely by-product of long-lasting interrogation sessions—on the likelihood of false confessions. Results confirm the hypothesis that individuals experiencing social exclusion are more likely to confess than individuals who are not socially excluded. These results suggest that situational variables in interrogations (specifically ostracism and exclusion) that emphasize important personality considerations (such as the need to belong) are important for police organizations to consider reducing the likelihood of false confessions. [ABSTRACT FROM AUTHOR]

Pathogenic intestinal bacteria lead to significant disease in humans. Here we investigated the role of the multifunctional protein, Apurinic/apyrimidinic endonuclease 1 (APE1), in regulating the internalization of bacteria into the intestinal epithelium. Intestinal tumor-cell lines and primary human epithelial cells were infected with Salmonella enterica serovar Typhimurium or adherent-invasive Escherichia coli. The effects of APE1 inhibition on bacterial internalization, the regulation of Rho GTPase Rac1 as well as the epithelial cell barrier function were assessed. Increased numbers of bacteria were present in APE1-deficient colonic tumor cell lines and primary epithelial cells. Activation of Rac1 was augmented following infection but negatively regulated by APE1. Pharmacological inhibition of Rac1 reversed the increase in intracellular bacteria in APE1-deficient cells whereas overexpression of constitutively active Rac1 augmented the numbers in APE1-competent cells. Enhanced numbers of intracellular bacteria resulted in the loss of barrier function and a delay in its recovery. Our data demonstrate that APE1 inhibits the internalization of invasive bacteria into human intestinal epithelial cells through its ability to negatively regulate Rac1. This activity also protects epithelial cell barrier function.

The emergence of the concept of religious moderation initiated by the Ministry of Religion of the Republic of Indonesia and set forth in the 2020-2024 National Medium-Term Development Plan (RPJMN) indicates that the program for internalizing and strengthening the values of religious moderation is very important and urgent to implement. The values of religious moderation consist of four points, namely national commitment, tolerance, anti-radicalism, and accommodative to local culture. The direction to internalize the four values of religious moderation is also played by the Anak Jalanan At-Tamur Islamic Boarding School, Bandung. This article aims to reveal the role of the Anak Jalanan At-Tamur Islamic Boarding School in internalizing the values of religious moderation to students. This article uses qualitative research methods that are field research. The research data were obtained from two sources, namely field data sources and library data sources. Data collection techniques used are observation, interviews, and documentaries. The data analysis techniques used are data condensation, data presentation, and drawing conclusions. The results showed that success in internalizing the values of religious moderation to students could be seen from the indicators or values of religious moderation. The four indicators (values) of religious moderation are reflected in the routine activities and attitudes of students of the Anak Jalanan At-Tamur Islamic Boarding School. Commitment to nationalism, tolerance, anti-radicalism and accommodativeness to local culture is imprinted in the personalities of the students of the Anak Jalanan At-Tamur Islamic Boarding School. This is also due to two things: firstly, interfaith dialogue events are often held and secondly, the leader of the Anak Jalanan At-Tamur Islamic Boarding School who has a broad and open view and emphasizes morality in his education.

Abstract Background The viral G-protein-coupled receptor (vGPCR) BILF1 encoded by the Epstein–Barr virus (EBV) is an oncogene and immunoevasin and can downregulate MHC-I molecules at the surface of infected cells. MHC-I downregulation, which presumably occurs through co-internalization with EBV-BILF1, is preserved among BILF1 receptors, including the three BILF1 orthologs encoded by porcine lymphotropic herpesviruses (PLHV BILFs). This study aimed to understand the detailed mechanisms of BILF1 receptor constitutive internalization, to explore the translational potential of PLHV BILFs compared with EBV-BILF1. Methods A novel real-time fluorescence resonance energy transfer (FRET)-based internalization assay combined with dominant-negative variants of dynamin-1 (Dyn K44A) and the chemical clathrin inhibitor Pitstop2 in HEK-293A cells was used to study the effect of specific endocytic proteins on BILF1 internalization. Bioluminescence resonance energy transfer (BRET)-saturation analysis was used to study BILF1 receptor interaction with β-arrestin2 and Rab7. In addition, a bioinformatics approach informational spectrum method (ISM) was used to investigate the interaction affinity of BILF1 receptors with β-arrestin2, AP-2, and caveolin-1. Results We identified dynamin-dependent, clathrin-mediated constitutive endocytosis for all BILF1 receptors. The observed interaction affinity between BILF1 receptors and caveolin-1 and the decreased internalization in the presence of a dominant-negative variant of caveolin-1 (Cav S80E) indicated the involvement of caveolin-1 in BILF1 trafficking. Furthermore, after BILF1 internalization from the plasma membrane, both the recycling and degradation pathways are proposed for BILF1 receptors. Conclusions The similarity in the internalization mechanisms observed for EBV-BILF1 and PLHV1-2 BILF1 provide a foundation for further studies exploring a possible translational potential for PLHVs, as proposed previously, and provides new information about receptor trafficking. Graphical Abstract

This study explains the phenomenon of the living qur'an practiced by the Tebuwung Society in Dukun, Gresik, East Java, regarding the meaning of surah Yusuf and Maryam in the tingkepantradition. The Qur'anic phenomenon in this pregnancy ritual is a form of meaning of the Qur'an's content that surrounds the local community's life. Based on qualitative research with a phenomenological approach, while data were obtained through in-depth interviews, observation, and documentation, this study tries to explain the phenomenon of the tingkepantradition, the public's reception of surahYusuf and Maryam, and the influence of reading the two surahsin the tingkepantradition. Based on Karl Mannheim's perspective and the social construction of Peter L. Berger, the study's results show that the awareness of the Qur'an that exists in the community is built through the religious understanding of the central figures of religion at local level. From these central figures, awareness emerged from understanding the ideas and moral messages of surahYusuf and Maryam in the hope of having descendants that handsome or beautiful face and character as depicted in the two surahs. Apart from being local wisdom, this tingkepan tradition constructively demonstrates the community's belief that the creation of man is closely related to the Oneness of God since he/she was in the womb.

The vegetative insecticidal proteins (Vip3A) secreted by some Bacillus thuringiensis (Bt) strains during vegetative growth are regarded as a new generation of insecticidal toxins. Like insecticidal crystal proteins, they are also used in transgenic crops to control pests. However, their insecticidal mechanisms are far less defined than those of insecticidal crystal protein. Prohibitin 2 (PHB2) is a potential Vip3Aa binding receptor identified from the membrane of Sf9 cells in our previous work. In this paper, we demonstrated the interaction between Vip3Aa and PHB2 using pull-down, dot blotting, microscale thermophoresis, and co-immunoprecipitation assays. PHB2 is distributed on the cell membrane and in the cytoplasm, and the co-localization of PHB2 and Vip3Aa was observed in Sf9 cells using a confocal laser scanning microscope. Moreover, PHB2 could interact with scavenger receptor-C via its SPFH (stomatin, prohibitin, flotillin, and HflK/C) domain. Downregulation of phb2 expression reduced the degree of internalization of Vip3Aa, exacerbated Vip3Aa-mediated mitochondrial damage, and increased Vip3Aa toxicity to Sf9 cells. This suggested that PHB2 performs two different functions: Acting as an interacting partner to facilitate the internalization of Vip3Aa into Sf9 cells and maintaining the stability of mitochondria. The latter has a more important influence on the virulence of Vip3Aa.

The imaging of different intercellular regions is attracting growing interest in the fields of biosensing, drug delivery, and gene therapy for cancer treatment. We present the synthesis of luminescent gold nanoclusters (AuNCs), grafted with a PEGylated U11 peptide derivative, for tumor recognition and their capacity to label the nucleus of pancreatic cancer cells. The short peptide named C3E6U11, used to synthesize and functionalize gold nanoclusters, is composed of a tricysteine sequence with side-chain thiol residues as peptide anchors for the Au nanoclusters together with the recognition peptide domain (U11). The active targeting red- or blue-emitting AuNCs can label pancreatic cancer cells and localize preferentially in the nucleus. The presence of U11 at their surface facilitates their penetration into pancreatic cell nuclei. Finally, in vivo imaging experiments in early embryos and free-swimming juvenile zebrafish are also performed to evaluate their biodistribution. [ABSTRACT FROM AUTHOR]

Streptococcus pyogenes causes a multitude of local and systemic infections, the most common being pharyngitis in children. Recurrent pharyngeal infections are common and are thought to be due to the re-emergence of intracellular GAS upon completion of antibiotic treatment. The role of colonizing biofilm bacteria in this process is not fully clear. Here, live respiratory epithelial cells were inoculated with broth-grown or biofilm bacteria of different M-types, as well as with isogenic mutants lacking common virulence factors. All M-types tested adhered to and were internalized into epithelial cells. Interestingly, internalization and persistence of planktonic bacteria varied significantly between strains, whereas biofilm bacteria were internalized in similar and higher numbers, and all strains persisted beyond 44 hours, showing a more homogenous phenotype. The M3 protein, but not the M1 or M5 proteins, was required for optimal uptake and persistence of both planktonic and biofilm bacteria inside cells. Moreover, the high expression of capsule and SLO inhibited cellular uptake and capsule expression was required for intracellular survival. Streptolysin S was required for optimal uptake and persistence of M3 planktonic bacteria, whereas SpeB improved intracellular survival of biofilm bacteria. Microscopy of internalized bacteria showed that planktonic bacteria were internalized in lower numbers as individual or small clumps of bacteria in the cytoplasm, whereas GAS biofilm bacteria displayed a pattern of perinuclear localization of bacterial aggregates that affected actin structure. Using inhibitors targeting cellular uptake pathways, we confirmed that planktonic GAS mainly uses a clathrin-mediated uptake pathway that also required actin and dynamin. Clathrin was not involved in biofilm internalization, but internalization required actin rearrangement and PI3 kinase activity, possibly suggesting macropinocytosis. Together these results provide a better understanding of the potential mechanisms of uptake and survival of various phenotypes of GAS bacteria relevant for colonization and recurrent infection. [ABSTRACT FROM AUTHOR]

Economic efficiency is the ultimate goal of all markets, including the electricity market. Several technical and pecuniary restrictions known as externalities in economics literature can significantly affect the economic efficiency of the electricity market. Negative externalities resulting from the operational restrictions of generation units are inherent to electricity markets. In this paper, after reviewing the effects of externalities on the day‐ahead electricity markets' economic efficiency using a unit commitment‐based model, an innovative and theoretically efficient service‐based procedure aimed at internalizing negative externalities in the day‐ahead electricity markets is presented. In this way, a new service procured by the energy storage system to provide energy interchange possibilities in the electricity market is introduced. The proposed service uses both price and quantity adjustment methods to internalize externalities. A new discriminatory method for pricing the service and a bi‐level optimization problem for determining the capacity of the energy storage system required to provide the service are considered. The consideration of the proposed method facilitates reaching the first‐best optimal market solution by alleviating negative externalities existing in the sub‐optimal second‐best solution in the presence of generation sector operational constraints. Numerical case studies demonstrate the functioning of the proposed externalities internalization scheme. [ABSTRACT FROM AUTHOR]

Silver nanoparticles (Ag-NPs) are widely used in daily life because of their antibacterial properties. A fraction of Ag-NPs are released into the ecosystem during their production and utilization. The toxicity of Ag-NPs has been reported. However, it is still disputed whether the toxicity is mainly due to the released silver ions (Ag+). In addition, few studies have reported the response of algae to metal nanoparticles under modulation of nitric oxide (NO). In this study, Chlorella vulgaris (C. vulgaris) was used as a model organism to study the toxic effects of Ag-NPs and Ag+ released from Ag-NPs on algae under the modulation of NO. The results showed that the biomass inhibition rate of Ag-NPs (44.84%) to C. vulgaris was higher than that of Ag+ (7.84%). Compared with Ag+, Ag-NPs induced more severe damage to photosynthetic pigments, photosynthetic system II (PSII) performance, and lipid peroxidation. More serious damage to cell permeability led to higher internalization of Ag under Ag-NPs stress. Application of exogenous NO reduced the inhibition ratio of photosynthetic pigments and chlorophyll autofluorescence. Further, NO reduced the MDA levels by scavenging reactive oxygen species induced by Ag-NPs. NO modulated the secretion of extracellular polymers and hampered the internalization of Ag. All these results showed that NO alleviates the toxicity of Ag-NPs to C. vulgaris. However, NO did not improve the toxic effects of Ag+. Our results provide new insights into the toxicity mechanism of Ag-NPs to algae modulated by the signal molecule NO. [ABSTRACT FROM AUTHOR]

Purpose: Angiotensin-(1–12) [Ang-(1–12)] serves as a primary substrate to generate angiotensin II (Ang II) by angiotensin-converting enzyme and/or chymase suggests it may be an unrecognized source of Ang II-mediated microvascular complication in hypertension-mediated retinopathy. We investigated Ang-(1–12) expression and internalization in adult retinal pigment epithelial-19 (ARPE-19) cultured cells. We performed the internalization of Ang-(1–12) in ARPE-19 cells in the presence of a highly specific monoclonal antibody (mAb) developed against the C-terminal end of the Ang-(1–12) sequence. Methods: All experiments were performed in confluent ARPE-19 cells (passage 28–35). We employed high-performance liquid chromatography to purify radiolabeled, 125I-Ang-(1–12) and immuno-neutralization with Ang-(1–12) mAb to demonstrate Ang-(1–12)'s internalization in ARPE-19 cells. Internalization was also demonstrated by immunofluorescence (IF) method. Results: These procedures revealed internalization of an intact 125I-Ang-(1–12) in ARPE-19 cells. A significant reduction (∼53%, P < 0.0001) in 125I-Ang-(1–12) internalization was detected in APRE-19 cells in the presence of the mAb. IF staining experiments further confirms internalization of Ang-(1–12) into the cells from the extracellular culture medium. No endogenous expression was detected in the ARPE-19 cells. An increased intensity of IF staining was detected in cells exposed to 1.0 μM Ang-(1–12) compared with 0.1 μM. Furthermore, we found hydrolysis of Ang-(1–12) into Ang II by ARPE-19 cells' plasma membranes. Conclusions: Intact Ang-(1–12) peptide is internalized from the extracellular spaces in ARPE-19 cells and metabolized into Ang II. The finding that a selective mAb blocks cellular internalization of Ang-(1–12) suggests alternate therapeutic approaches to prevent/reduce the RPE cells Ang II burden. [ABSTRACT FROM AUTHOR]

Copyright of Journal of China Tourism Research is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spreads efficiently by spike-mediated, direct cell-to-cell transmission. However, the underlying mechanism is poorly understood. Herein, we demonstrate that the tight junction protein occludin (OCLN) is critical to this process. SARS-CoV-2 infection alters OCLN distribution and expression and causes syncytium formation that leads to viral spread. OCLN knockdown fails to alter SARS-CoV-2 binding but significantly lowers internalization, syncytium formation, and transmission. OCLN overexpression also has no effect on virus binding but enhances virus internalization, cell-to-cell transmission, and replication. OCLN directly interacts with the SARS-CoV-2 spike, and the endosomal entry pathway is involved in OCLN-mediated cell-to-cell fusion rather than in the cell surface entry pathway. All SARS-CoV-2 strains tested (prototypic, alpha, beta, gamma, delta, kappa, and omicron) are dependent on OCLN for cell-to-cell transmission, although the extent of syncytium formation differs between strains. We conclude that SARS-CoV-2 utilizes OCLN as an internalization factor for cell-to-cell transmission. [ABSTRACT FROM AUTHOR]

Although amoxicillin (AMPC) is recommended as first-line therapy for acute pharyngotonsillitis caused by group A streptococci (GAS), it often fails to eradicate infections. Internalization and subsequent intracellular survival of GAS are considered major mechanisms for penicillin therapeutic failure. It is, therefore, desirable to administer drugs that exert bactericidal effects on extracellular and intracellular GAS. In this study, we aim to investigate the bactericidal effects of lascufloxacin (LSFX) on internalized GAS in HEp-2 cells. The GAS strain M1 and clinical isolate strain #2 were used in this study. Following treatment of GAS-infected human pharyngeal carcinoma epithelial HEp-2 cells with LSFX or AMPC, internalized GAS cells were recovered. The concentrations of LSFX and AMPC were equivalent to 1 × and 2 × MIC for strain M1. Culture medium was used as a control. Time-lapse and fluorescence images of GAS invading HEp-2 cell were obtained. LIVE/DEAD fluorescence staining was used to confirm the viability of internalized GAS. LSFX significantly reduced the number of cell-internalized M1 and #2 GAS strains compared to the control (p < 0.01) in a dose-dependent manner. However, AMPC did not reduce this in both strains. Both live and dead intracellular GAS were confirmed in HEp-2 cells exposed to LSFX. In contrast, intracellular GAS survived in HEp-2 cells exposed to AMPC and in the control. LSFX elicits significant bactericidal effects on cell-internalized GAS, hence it may represent a potent therapeutic option for patients with acute pharyngotonsillitis in whom AMPC treatment has failed. [ABSTRACT FROM AUTHOR]

Coronavirus membrane protein is a major component of the viral envelope and plays a central role in the viral life cycle. Studies of the coronavirus membrane protein (M) have mainly focused on its role in viral assembly and budding, but whether M protein is involved in the initial stage of viral replication remains unclear. In this study, eight proteins in transmissible gastroenteritis virus (TGEV)-infected cells coimmunoprecipitated with monoclonal antibodies (MAb) against M protein in PK-15 cells, heat shock cognate protein 70 (HSC70), and clathrin were identified by matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry (MALDI-TOF MS). Further studies demonstrated that HSC70 and TGEV M colocalized on the cell surface in early stages of TGEV infection; specifically, HSC70 bound M protein through its substratebinding domain (SBD) and preincubation of TGEV with anti-M serum to block the interaction of M and HSC70 reduced the internalization of TGEV, thus demonstrating that the M-HSC70 interaction mediates the internalization of TGEV. Remarkably, the process of internalization was dependent on clathrin-mediated endocytosis (CME) in PK-15 cells. Furthermore, inhibition of the ATPase activity of HSC70 reduced the efficiency of CME. Collectively, our results indicated that HSC70 is a newly identified host factor involved in TGEV infection. Taken together, our findings clearly illustrate a novel role for TGEV M protein in the viral life cycle and present a unique strategy used by HSC70 to promote TGEV infection in which the interaction with M protein directs viral internalization. These studies provide new insights into the life cycle of coronaviruses. IMPORTANCE TGEV is the causative agent of porcine diarrhea, a viral disease that economically affects the pig industry in many countries. However, the molecular mechanisms underlying viral replication remain incompletely understood. Here, we provide evidence of a previously undescribed role of M protein in viral replication during early stages. We also identified HSC70 as a new host factor affecting TGEV infection. We demonstrate that the interaction between M and HSC70 directs TGEV internalization in a manner dependent on CME, thus revealing a novel mechanism for TGEV replication. We believe that this study may change our understanding of the first steps of infection of cells with coronavirus. This study should facilitate the development of anti-TGEV therapeutic agents by targeting the host factors and may provide a new strategy for the control of porcine diarrhea. [ABSTRACT FROM AUTHOR]

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The aim of this research was to highlight the importance of internalizing Christian religious doctrine in strengthening Christian identity, due to the declining faith among Indonesian Christian youth, which was reflected in the increasing number of members exiting from the church. Therefore, this research attempted to strengthen the identity of Indonesian Christian youth by developing what is termed the Moses internalization model. Social research methods and hermeneutics were used in order to achieve appropriate results. The results could be used to develop the Moses internalization model in order to teach Christian religious doctrine to young Indonesians in churches and schools, thereby strengthening their level of identity. The paper demonstrates the centrality of the example posed by Moses which serves as an internalization process, which has four stages. The first is doctrinal information, the second stage comprises doctrinal application exercises or training, the third is identity formation, and the fourth stage is the crystallization and strengthening of identity. Moses informed the Israelites through doctrines that God is One and God is God. The people were to fear the Lord and keep all the given statutes and commandments. They were taught to love the Lord God with all their heart, soul, and strength (Deuteronomy 6:4-5). The model or framework of Moses for a Christian's identity can be applied to pastoral ministry. This model is thus is offered as a support for pastors in the important aspect of forging and internalizing strong Christian identities in their adherents. [ABSTRACT FROM AUTHOR]

Neuropathic pain (NP) refers to pain caused by primary or secondary damage or dysfunction of the peripheral or central nervous system, which seriously affects the physical and mental health of 7–10% of the general population. The etiology and pathogenesis of NP are complex; as such, NP has been a hot topic in clinical medicine and basic research for a long time, with researchers aiming to find a cure by studying it. Opioids are the most commonly used painkillers in clinical practice but are regarded as third-line drugs for NP in various guidelines due to the low efficacy caused by the imbalance of opioid receptor internalization and their possible side effects. Therefore, this literature review aims to evaluate the role of the downregulation of opioid receptors in the development of NP from the perspective of dorsal root ganglion, spinal cord, and supraspinal regions. We also discuss the reasons for the poor efficacy of opioids, given the commonness of opioid tolerance caused by NP and/or repeated opioid treatments, an angle that has received little attention to date; in-depth understanding might provide a new method for the treatment of NP. [ABSTRACT FROM AUTHOR]

As the climate worsens and the demand for food grows, so does the interest in nanoagriculture. The interaction between plants and nanomaterials (NMs) has been extensively and intensively examined. However, stopping at the outcome of a phenomenon is often insufficient. Therefore, we introduce three important processes of nanoparticleplant interactions: translocation, transformation, and plant metabolism. During the migration of nanoparticles, size and surface electrical properties are the main determining factors. Additionally, the interaction of nanoparticles with cell membranes is another key aspect of research. The transformation of nanoparticles in plants is mainly due to redox substances. The way that nanoparticles affect plant metabolism may be able to shed light on the interaction of nanoparticles with plants. This review adds to the existing knowledge on the design of nanoagrochemicals and summarizes the mechanism of interaction of NMs with plants. In this way, NMs can be used for their beneficial effects and thus contribute to the maintenance of food security and sustainable development. [ABSTRACT FROM AUTHOR]

Pathogenic mycobacteria gain entry to their hosts by inhalation or ingestion where they adhere to different cell types and are subsequently internalized by professional phagocytic cells, such as macrophages or dendritic cells. Multiple pathogen-associated molecular patterns present on the mycobacterial surface are recognized by and interact with a diverse panel of phagocytic pattern recognition receptors, representing the first step of the infection process. This review summarizes the current knowledge on the numerous host cell receptors and their associated mycobacterial ligands or adhesins. It further discusses the downstream molecular and cellular events resulting from the engagement of the various receptor-mediated pathways, leading to either intracellular survival of mycobacteria or to activation of host immune defenses. The content presented herein on adhesins and host receptors may serve as a resource for those developing novel therapeutic approaches, e.g. in the design of antiadhesin molecules to prevent bacterial attachment and infection. The collection of mycobacterial surface molecules highlighted in this review may also provide potential new therapeutic targets, diagnostic markers, or vaccine candidates to combat these notoriously challenging and persistent pathogens. [ABSTRACT FROM AUTHOR]

It is well-established that double-stranded RNA (dsRNA) exhibits noticeable radioprotective and radiotherapeutic effects. The experiments conducted in this study directly demonstrated that dsRNA was delivered into the cell in its native form and that it induced hematopoietic progenitor proliferation. The 68 bp synthetic dsRNA labeled with 6-carboxyfluorescein (FAM) was internalized into mouse hematopoietic progenitors, c-Kit+ (a marker of long-term hematopoietic stem cells) cells and CD34+ (a marker of short-term hematopoietic stem cells and multipotent progenitors) cells. Treating bone marrow cells with dsRNA stimulated the growth of colonies, mainly cells of the granulocyte–macrophage lineage. A total of 0.8% of Krebs-2 cells internalized FAM-dsRNA and were simultaneously CD34+ cells. dsRNA in its native state was delivered into the cell, where it was present without any signs of processing. dsRNA binding to a cell was independent of cell charge. dsRNA internalization was related to the receptor-mediated process that requires energy from ATP. Synthetic dsRNA did not degrade in the bloodstream for at least 2 h. Hematopoietic precursors that had captured dsRNA reinfused into the bloodstream and populated the bone marrow and spleen. This study, for the first time, directly proved that synthetic dsRNA is internalized into a eukaryotic cell via a natural mechanism. [ABSTRACT FROM AUTHOR]

Nanomaterials offer a broad spectrum of applications in biomedicine. The shapes of gold nanoparticles could modulate tumor cell behavior. Spherical (AuNPsp), stars (AuNPst) and rods (AuNPr) shapes of polyethylene glycol coated-gold nanoparticles (AuNPs-PEG) were synthesized. Metabolic activity, cellular proliferation, and reactive oxygen species (ROS) were measured and the impact of AuNPs-PEG in metabolic enzymes function was evaluated by RT-qPCR in PC3, DU145, and LNCaP prostate cancer cells. All AuNPs were internalized, and the different morphologies of AuNPs showed to be an essential modulator of metabolic activity. For PC3 and DU145, the metabolic activity of AuNPs was found to rank in the following order from lowest to highest: AuNPsp-PEG, AuNPst-PEG, and AuNPr-PEG. Regarding LNCaP cells, the AuNPst-PEG were less toxic, followed by AuNPsp-PEG and AuNPr-PEG, but it seems not to be dose-dependent. The proliferation was lower in AuNPr-PEG in PC3 and DU145 cells but was stimulated around 10% in most conditions (0.001–0.1 mM) in LNCaP cells (not statistically significant). For 1 mM, LNCaP cells showed a significant decrease in proliferation only for AuNPr-PEG. The outcomes of the current study demonstrated that different AuNPs conformations influence cell behavior, and the correct size and shape must be chosen considering its final application in the field of nanomedicine. [ABSTRACT FROM AUTHOR]

The purpose of this current study is to explore the effect advertisements have on the body image and self-esteem of women. Over the years, women have been represented in a stereotypical manner in advertisements which are largely focused on beauty ideals and the thin-ideal. This can significantly affect women's body image perceptions and self-esteem. This study aims to investigate how the perceptions of body image and self-esteem of women are affected by the depiction of appearance ideals, regarding women, in advertisements. A survey was conducted among 216 women between the ages of 18 to 30 years old in Pakistan. It was found that advertisements depicting appearance ideals do effect women's body image and self-esteem; as well as, lead to the internalization of sociocultural ideals regarding appearance. This study found that advertisements caused body dissatisfaction and greater internalization of sociocultural ideals in women. However, women in this study reported higher levels of self-esteem, contrary to other researches. This has been attributed to the increased diversity in the representation of appearance ideals and exposure to moderate-size models. In conclusion, this research confirms the significant effect advertisements have on how young Pakistani women perceive their body image and self-worth. [ABSTRACT FROM AUTHOR]