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Most literature on China and the liberal international order has described how China behaves towards the multiple elements of the order, but virtually no systematic efforts were undertaken to understand the rationale or motivation of that behaviour. Hence, such studies do not clarify whether compliance comes from an actual commitment to the order's ideas or if it is instrumental in order to acquire legitimacy and avoid social sanctions. To address this gap in literature, this article elaborates on the concept of internalization. I separate partial from full internalization: both require substantial compliance with norms, but in the former states halt compliance when interests are at stake. I identify Chinese interests to, then, assess if China internalized the three main institutions of the liberal international order: sovereignty, human rights and the market. I conclude that none of them was fully internalized yet, although there is partial internalization of sovereignty and the market. This means that China's compliance with the order's norms is higher than many assume, but that it is selective and subordinated to Chinese core interests. [ABSTRACT FROM AUTHOR]

Medium of instruction has been shifting towards English, thereby producing English as a medium of instruction (EMI) in Saudi higher education due to internationalization, globalization, economic and technological advancement. Saudi universities have been adopting EMI to address the needs of the day and the demands by the stakeholders and students. This study explores the perceptions of Saudi university students and teachers about the use of EMI in the Saudi higher education. The participants are 80 students and 20 university teachers affiliated to Prince Sattam bin Abdulaziz University, Kingdom of Saudi Arabia. A fivepoint Likert-scale questionnaire was used to collect data. The findings show that both teachers and students hold positive perceptions about the use of EMI in Saudi universities. The findings suggest that teachers hold stronger positive attitudes towards the use of EMI. Students demonstrate a positive attitudinal behavior when teachers switch to Arabic while teaching. Findings further show that the use of EMI may hamper the native language and culture as perceived by the students. A number of implications have been suggested as students may feel hesitant to speak in English due to their low English proficiency. [ABSTRACT FROM AUTHOR]

Melittin, as an agent to lyse biological membranes, may be a promising therapeutic agent in the treatment of cancer. However, because of its nonspecific actions, there is a need to use a delivery method. The conducted research determined whether carbon nanoparticles, such as graphene and graphene oxide, could be carriers for melittin to breast cancer cells. The studies included the analysis of intracellular pH, the potential of cell membranes, the type of cellular transport, and the expression of receptor proteins. By measuring the particle size, zeta potential, and FT-IT analysis, we found that the investigated nanoparticles are connected by electrostatic interactions. The level of melittin encapsulation with graphene was 86%, while with graphene oxide it was 78%. A decrease in pHi was observed for all cell lines after administration of melittin and its complex with graphene. The decrease in membrane polarization was demonstrated for all lines treated with melittin and its complex with graphene and after exposure to the complex of melittin with graphene oxide for the MDA-MB-231 and HFFF2 lines. The results showed that the investigated melittin complexes and the melittin itself act differently on different cell lines (MDA-MB-231 and MCF-7). It has been shown that in MDA-MD-231 cells, melittin in a complex with graphene is transported to cells via caveolin-dependent endocytosis. On the other hand, the melittin–graphene oxide complex can reach breast cancer cells through various types of transport. Other differences in protein expression changes were also observed for tumor lines after exposure to melittin and complexes. [ABSTRACT FROM AUTHOR]

A conventional consensus on organizational change is the momentum view, which claims that prior changes of a given kind increase the probability of a subsequent change of the same kind. This consensus has been recently challenged by the deceleration view, which argues that prior changes decrease the probability of a subsequent change. By making a distinction between experienced changes and internalized changes, this study postulates that the two views complement and coexist with each other. A change of a given kind is less likely to occur with more experienced changes of the same kind, while it is more likely to occur with more internalized changes of the same kind. This integrated view is supported in a sample of 477 U.S. local governments making decisions about whether to outsource public services during 1982–2007. This study also looks into how relatedness to internalized changes affected the occurrence of a subsequent change of the same kind. The impact of operational relatedness was positive at an early time and increasingly negative in the long run. The impact of skill relatedness, however, was positive and steadily increased as time went on. MAD statement An important debate on organizational change is how prior changes of a given kind affect the probability of a subsequent change of the same kind. Whereas the momentum view claims that prior changes breed a subsequent change, the deceleration view contends that prior changes prevent a subsequent change. By distinguishing between experienced and internalized changes, this study argues and demonstrates, with a sample of U.S. local governments, that the two views complement and coexist with each other. A change is less likely to occur with more experienced changes, while it is more likely to occur with more internalized changes. [ABSTRACT FROM AUTHOR]

Traditional storytelling has been an essential constituent of Indigenous cultures in North America, including contemporary literary production. The perception of storytelling in Indigenous theories might, however, in some aspects differ from its Western literary critical understanding. This paper outlines the conceptual frameworks which shape and are reflected in Indigenous storytelling and examines relationality as the core principle of Indigenous knowledge system. Relationality does not only represent a frequent theme in Indigenous works, but it is also manifested in their narrative structure, and simultaneously can be used for their interpretation. In addition, the paper focuses on the equal role of the storyteller and the listener in Indigenous views on storytelling, the necessity to internalize a story to derive meaning, the power and animacy of stories and the importance of this for Indigenous typologies of stories, and on the issue of cultural appropriation, which stems from the different approaches to stories and knowledge as such in Indigenous and non-Indigenous perceptions.

The article analyzes the education system role in providing the effective scienceFinnovation process in modern Russia and in the aspect of future transformation to science economy. The authors assert that the potential of education system in research and development is restrictedinasmuch as its primary function is to create the educational environment. PreFeminently, itis able to ensure an appropriate level of innovative manpower. The key role of education in organizing the interaction between science and industry, and in formation of innovation infrastructure is substantiated. The education system is also viewed as the main factor in the knowledge management systems organization.

Exposure to VEGF-A 165 a over several days leads to a persistent dysfunction of the very tight barrier formed by immortalized endothelial cells of the bovine retina (iBREC). Elevated permeability of the barrier is indicated by low cell index values determined by electric cell-substrate impedance measurements, by lower amounts of claudin-1, and by disruption of the homogenous and continuous staining of vascular endothelial cadherin at the plasma membrane. Because of findings that suggest modulation of VEGF-A's detrimental effects on the inner blood-retina barrier by the angiogenic growth factor angiopoietin-2, we investigated in more detail in vitro whether this growth factor indeed changes the stability of the barrier formed by retinal endothelial cells or modulates effects of VEGF-A. In view of the clinical relevance of anti-VEGF therapy, we also studied whether blocking VEGF-A-driven signaling is sufficient to prevent barrier dysfunction induced by a combination of both growth factors. Although angiopoietin-2 stimulated proliferation of iBREC, the formed barrier was not weakened at a concentration of 3 nM: Cell index values remained high and expression or subcellular localization of claudin-1 and vascular endothelial cadherin, respectively, were not affected. Angiopoietin-2 enhanced the changes induced by VEGF-A 165 a and this was more pronounced at lower concentrations of VEGF-A 165 a. Specific inhibition of the VEGF receptors with tivozanib as well as interfering with binding of VEGF-A to its receptors with bevacizumab prevented the detrimental effects of the growth factors; dual binding of angiopoietin-2 and VEGF-A by faricimab was marginally more efficient. Uptake of extracellular angiopoietin-2 by iBREC can be efficiently prevented by addition of faricimab which is also internalized by the cells. Exposure of the cells to faricimab over several days stabilized their barrier, confirming that inhibition of VEGF-A signaling is not harmful to this cell type. Taken together, our results confirm the dominant role of VEGF-A 165 a in processes resulting in increased permeability of retinal endothelial cells in which angiopoietin-2 might play a minor modulating role. • Angiopoietin-2 does not impair the barrier formed by retinal endothelial cells. • Angiopoietin-2 amplifies barrier impairment induced by VEGF-A. • Inhibiting VEGF signaling prevents barrier impairment induced by VEGF-A plus ANG2. • Dual binding of VEGF-A and ANG2 by faricimab is not more efficient. • Faricimab prevents internalization of ANG2 but not of VEGF-A. [ABSTRACT FROM AUTHOR]

[Display omitted] • Nuclear localization sequence (NLS) tagged with HER2-targeting rL-A9 peptide. • Confocal microscopy assessed significant cellular internalization of NLS-tagged peptide. • High binding and cellular internalization observed for [177Lu]Lu-DOTA-NLS-rL-A9. • [177Lu]Lu-DOTA-NLS-rL-A9 exhibited consistent tumor retention (48 h) with negligible washout of radioactivity. • Significantly high tumor-to-background ratios observed for [177Lu]Lu-DOTA-NLS-rL-A9. Tagging of cell permeable nuclear localization sequence (NLS) with receptor targeting peptide vectors is an attractive strategy for selectively targeted translocation of therapeutic cargoes. The present study aimed at grafting nuclear localization sequence (NLS) onto breast cancer targeting rL-A9 peptide. Molecular docking analysis revealed higher binding affinity of the peptide, DOTA-NLS-rL-A9 (−26.1 kJ/mol) towards HER2 receptor in comparison to DOTA-rL-A9 peptide (−22.2 kJ/mol). Confocal microscopy data suggested significantly enhanced cellular internalization of NLS-tagged peptide. The engineered HER2-selective, DOTA-NLS-rL-A9 peptide scaffold was radiolabeled with Lu-177 for intracellular delivery of the theranostic radionuclide into tumor cells. [177Lu]Lu-DOTA-NLS-rL-A9 exhibited significantly enhanced binding affinity (4.58 ± 1.77 nM) towards human breast carcinoma SKBR3 cells and cellular internalization (85 % at 24 h) compared to its original analog, [177Lu]Lu-DOTA-rL-A9. In vivo biodistribution studies showed consistent retention of [177Lu]Lu-DOTA-NLS-rL-A9 in the tumor with negligible washout of radioactivity (∼4.1 % ID/g at 48 h). Prolonged tumor activity with rapid off-target tissue clearance resulted in significantly high tumor-to-background ratios. The radiopeptide, [177Lu]Lu-DOTA-NLS-rL-A9 thus, being precisely confined into HER2-expressing tumor cells and exhibiting favourable pharmacokinetic features is an efficient candidate for further screening. [ABSTRACT FROM AUTHOR]

Signaling through classical death receptor Fas was mainly appreciated as a pro-death pathway until recent reports characterized pro-inflammatory outcomes of Fas-mediated activation in pathological contexts. How Fas signaling can switch to pro-inflammatory activation is poorly understood. Herein, we report that in macrophages and neutrophils, the Toll-like receptor (TLR) adapter CD14 determines the inflammatory output of Fas-mediated signaling. Our findings propose CD14 as a crucial chaperone of Fas receptor internalization in macrophages and neutrophils, resulting in Cd14 −/− myeloid cells that are protected from FasL-induced apoptosis, activate nuclear factor κB (NF-κB), and release cytokines in response. As in TLR signaling, CD14 is also required for Fas to signal through the adaptor TRIF (TIR-domain-containing adapter-inducing interferon-β) and induce a pro-death complex. Our findings demonstrate that CD14 availability can determine the switch between Fas-mediated pro-death and pro-inflammatory outcomes by internalizing the receptor. [Display omitted] • Fas-mediated death of myeloid cells requires TLR-associated protein CD14 TRIF • Without CD14, Fas activation induces NF-κB-mediated cytokine production in vitro and in vivo • CD14 promotes internalization of Fas from the cell surface and formation of a death complex Activation of the Fas receptor can be pro-death or pro-inflammatory, but little is known about the switch between outcomes. Magri et al. demonstrate that TLR adaptor CD14 can mediate this switch between death and inflammation in myeloid cells by internalizing the Fas receptor to a pro-death complex and preventing inflammation. [ABSTRACT FROM AUTHOR]

[Display omitted] • New A56 -based PROTACs target-degrading PD-L1 have been designed and synthesized. • Compounds 9i and 9j induce the internalization and degradation of PD-L1 in HCC-827 cells. • Compounds 9i and 9j degrade of PD-L1 through dual means of lysosome and proteasome. • The compounds have good anti-tumor activity by activating immunity in vivo. Currently, antibody drugs targeting programmed cell death ligand 1 (PD-L1) have achieved promising results in cancer treatment, while the development of small-molecule drugs lags behind. In this study, we designed and synthesized a series of PD-L1-degrading agents based on the PROTAC design principle, utilizing the PD-L1 inhibitor A56. Through systematic screening of ligands and linkers and investigating the structure–activity relationship of the degraders, we identified two highly active compounds, 9i and 9j. These compounds enhance levels of CD4+, CD8+, granzyme B, and perforin, demonstrating significant in vivo antitumor effects with a tumor growth inhibition (TGI) of up to 57.35 %. Both compounds facilitate the internalization of PD-L1 from the cell surface and promote its degradation through proteasomal and lysosomal pathways, while also maintaining inhibition of the PD-1/PD-L1 interaction. In summary, our findings provide a novel strategy and mechanism for developing biphenyl-based PROTAC antitumor drugs targeting and degrading PD-L1. [ABSTRACT FROM AUTHOR]

The internalization of using resources is meant to empower the local economy, and under the current crisis circumstances it could help as a powerful tool to the recovery of the rural sector. From the farming point of view, the cutting cost policy is the main theoretical justification for internalization of actives, and by extension, of activities, and in the European case it concerns the entire continental economic system. Internalization emphasizes the protectionist economic activities, mainly in line with anti-crisis measures for guiding markets. This persisting discrepancy among European states in terms of development and yearly performances emphasizes the necessity of empowering the internal efforts of our country in the field of production, and agricultural production. This is a new shape of introducing the dirigisme and the macroeconomic interventionism as a fundamental tool against the negative effects of the economic slowing-down, especially in zoo-economic activities, after many critics addressed to the centralized economic intervention.

Abstract Background Use of dietary supplements (DS) and muscle enhancing dietary supplements (MEDS) is frequent among students despite the lack of evidence of effects and health risks related to consumption. We need to increase our understanding of students’ motivation, examine potential gender differences, and explore explanatory factors, to address preventive measures related to use. Therefore, this study aimed to explore the frequency of, and the reasons for, DS use among university students, as well as explanatory factors for use of MEDS. Method Male and female students from nine Norwegian universities participated in this cross-sectional study. Participants responded to questions about demographics, DS and MEDS use, internalization of body ideals, physical activity- and exercise level, motives for exercise, and exercise context. Independent t-test, Chi-square test, Pearson’s correlation, and logistic regression were used to investigate between group differences, associations, and explanatory factors for use, respectively. P-values ≤ 0.05 were defined as significant. Results A total of 1001 males (34%) and females, with a mean(sd) age of 24.21(4.76) years, participated. The frequency of DS use was 42% and 40% (p = .414) in males and females, respectively, in which more males than females used DS to improve physical or mental performance (p =

This research aims to obtain the results of an analysis of how to internalize basic values in the business unit of the University of Darussalam Gontor in human resource development. This research was conducted using field research. The research was conducted by the Business Unit of The University of Darussalam Gontor, located on the Siman Highway, Siman Subdistrict, Ponorogo Regency, East Java Province. Informants in this study include 1 President Director, 2 Company Managers, 2 Staff who are also Students, and 1 Permanent Employee in the Business Unit of The University of Darussalam Gontor. In this study, the type of observation used passive participation means that researchers are not involved in the Business Unit activities of The University of Darussalam Gontor. Researchers conduct interviews with a free but controlled system, where the interview does freely what the interview will do to the interviewer but still leads to talks. The results showed that the implementation of internalization of basic values including Sincerity, Simplicity, Independence, Ukhuwah Islamiyah or Togetherness, and Freedom in the development of human resources Business Unit Darussalam Gontor University has been in accordance with the theory of internalization stages, internalization strategy, training and development methods and has a function to improve the moral and moral human resources. Further quantitative analysis is needed, such as analyzing influential variables to enhance the performance of the Business Unit of Universitas Darussalam Gontor.

Sepsis is defined as a life-threatening dysfunction due to a dysregulated host response to infection. It is a common and complex syndrome and is the leading cause of death in intensive care units. The lungs are most vulnerable to the challenge of sepsis, and the incidence of respiratory dysfunction has been reported to be up to 70%, in which neutrophils play a major role. Neutrophils are the first line of defense against infection, and they are regarded as the most responsive cells in sepsis. Normally, neutrophils recognize chemokines including the bacterial product N-formyl-methionyl-leucyl-phenylalanine (fMLP), complement 5a (C5a), and lipid molecules Leukotriene B4 (LTB4) and C-X-C motif chemokine ligand 8 (CXCL8), and enter the site of infection through mobilization, rolling, adhesion, migration, and chemotaxis. However, numerous studies have confirmed that despite the high levels of chemokines in septic patients and mice at the site of infection, the neutrophils cannot migrate to the proper target location, but instead they accumulate in the lungs, releasing histones, DNA, and proteases that mediate tissue damage and induce acute respiratory distress syndrome (ARDS). This is closely related to impaired neutrophil migration in sepsis, but the mechanism involved is still unclear. Many studies have shown that chemokine receptor dysregulation is an important cause of impaired neutrophil migration, and the vast majority of these chemokine receptors belong to the G protein-coupled receptors (GPCRs). In this review, we summarize the signaling pathways by which neutrophil GPCR regulates chemotaxis and the mechanisms by which abnormal GPCR function in sepsis leads to impaired neutrophil chemotaxis, which can further cause ARDS. Several potential targets for intervention are proposed to improve neutrophil chemotaxis, and we hope that this review may provide insights for clinical practitioners.

Cellular interactions between delta and mu opioid receptors (DORs and MORs), including heteromerization, are thought to regulate opioid analgesia. However, the identity of the nociceptive neurons in which such interactions could occur in vivo remains elusive. Here we show that DOR-MOR co-expression is limited to small populations of excitatory interneurons and projection neurons in the spinal cord dorsal horn and unexpectedly predominates in ventral horn motor circuits. Similarly, DOR-MOR co-expression is rare in parabrachial, amygdalar, and cortical brain regions processing nociceptive information. We further demonstrate that in the discrete DOR-MOR co-expressing nociceptive neurons, the two receptors internalize and function independently. Finally, conditional knockout experiments revealed that DORs selectively regulate mechanical pain by controlling the excitability of somatostatin-positive dorsal horn interneurons. Collectively, our results illuminate the functional organization of DORs and MORs in CNS pain circuits and reappraise the importance of DOR-MOR cellular interactions for developing novel opioid analgesics.

Human norovirus (NoV) is a leading cause of fresh produce associated outbreaks. Previous research indicates that the roots of growing leafy greens and berries internalize human NoV. However the effect of plant type and inoculum level on internalization rates has not been directly compared. In this study we compared the internalization and dissemination rates of human NoV and its surrogate, Tulane virus (TV) in green onion, radishes, and Romaine lettuce. We also evaluated the effect inoculum level and plant growth matrix on the rate of viral internalization. In the hydroponic growth system, we detected internalization and dissemination of human NoV RNA in green onions. In hydroponically growing green onions inoculated with high titer TV, we found higher rates of internalization and dissemination compared to green onions inoculated with low titer TV. In soil growth systems, no infectious TV was detected in either green onion or radishes. However, in Romaine lettuce plants grown in soil approximately 4 log10 PFU/g was recovered from all tissues on day 14 p.i. Overall, we found that the type of plant, growth matrix, and the inoculum level influences the internalization and dissemination of human NoV and TV.

Recently, we showed that HIV-1 is sequestered, i.e., trapped, in the intracellular vesicles of oral and genital epithelial cells. Here, we investigated the mechanisms of HIV-1 sequestration in vesicles of polarized tonsil, foreskin and cervical epithelial cells. HIV-1 internalization into epithelial cells is initiated by multiple entry pathways, including clathrin-, caveolin/lipid raft-associated endocytosis and macropinocytosis. Inhibition of HIV-1 attachment to galactosylceramide and heparan sulfate proteoglycans, and virus endocytosis and macropinocytosis reduced HIV-1 sequestration by 30-40%. T-cell immunoglobulin and mucin domain 1 (TIM-1) were expressed on the apical surface of polarized tonsil, cervical and foreskin epithelial cells. However, TIM-1-associated HIV-1 macropinocytosis and sequestration were detected mostly in tonsil epithelial cells. Sequestered HIV-1 was resistant to trypsin, pronase, and soluble CD4, indicating that the sequestered virus was intracellular. Inhibition of HIV-1 intraepithelial sequestration and elimination of vesicles containing virus in the mucosal epithelium may help in the prevention of HIV-1 mucosal transmission.

Peripheral endothelial cells are capable of erythrophagocytosis, but data on brain endothelial erythrophagocytosis are limited. We studied the relationship between brain endothelial erythrophagocytosis and cerebral microhemorrhage, the pathological substrate of MRI-demonstrable cerebral microbleeds. To demonstrate the erythrophagocytic capability of the brain endothelium, we studied the interactions between brain endothelial cells and red blood cells exposed to oxidative stress in vitro, and developed a new in vitro cerebral microbleeds model to study the subsequent passage of hemoglobin across the brain endothelial monolayer. Using multiple approaches, our results show marked brain endothelial erythrophagocytosis of red blood cells exposed to oxidative stress compared with control red blood cells in vitro. This brain endothelial erythrophagocytosis was accompanied by passage of hemoglobin across the brain endothelial monolayer with unaltered monolayer integrity. In vivo and confocal fluorescence microscopy studies confirmed the extravasation of RBC exposed to oxidative stress across brain endothelium. These findings, demonstrating erythrophagocytosis mediated by the brain endothelial monolayer and the subsequent passage of iron-rich hemoglobin in vitro and RBC in vivo, may have implications for elucidating mechanisms involved in the development of cerebral microbleeds that are not dependent on disruption of the microvasculature.

The uptake and trafficking of cell surface receptors can be monitored by a technique called 'antibody-feeding' which uses an externally applied antibody to label the receptor on the surface of cultured, live cells. Here, we adapt the traditional antibody-feeding experiment to polarized epithelial cells (Madin-Darby Canine Kidney) grown on permeable Transwell supports. By adding two tandem extracellular Myc epitope tags to the C-terminus of the SNARE protein syntaxin 3 (Stx3), we provided a site where an antibody could bind, allowing us to perform antibody-feeding experiments on cells with distinct apical and basolateral membranes. With this procedure, we observed the endocytosis and intracellular trafficking of Stx3. Specifically, we assessed the internalization rate of Stx3 from the basolateral membrane and observed the ensuing endocytic route in both time and space using immunofluorescence microscopy on cells fixed at different time points. For cell lines that form a polarized monolayer containing distinct apical and basolateral membranes when cultured on permeable supports, e.g., MDCK or Caco-2, this protocol can measure the rate of endocytosis and follow the subsequent trafficking of a target protein from either limiting membrane.

Several studies previously showed that the NFL-TBS.40–63 peptide (NFL-peptide) is capable to specifically penetrating several glioblastoma cell lines (rat, mouse, human) and inhibiting their cell division in vitro and their tumor development in vivo. When lipid nanocapsules (LNCs) are functionalized with the NFL-peptide, their absorption is targeted in glioblastoma cells both in vitro and in vivo. In the present study, we investigated the molecular architecture of these nanovectors (LNC-NFL) by using several microscopy techniques (transmission electron microscopy, cryo-electron microscopy, and cryo-electron tomography). We also used high-performance liquid chromatography (UPLC) technique to evaluate the interaction between LNCs and peptides. The work shows that the NFL-peptide forms stable long filaments along which the lipid nanocapsules interact strongly to form some sort of nanomolecular bracelets. This new construction composed of the NFL-peptide and lipid nanocapsules shows a better internalization in rat glioblastoma cells (F98 cells) than lipid nanocapsules alone.

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While automated feedback is becoming readily accessible to student writers, how students employ resources and strategies to use such feedback remains largely unexplored. Informed by activity theory and the construct of appropriation, this study conceptualizes students' use of automated feedback as social appropriation mediated by resources and internalization strategies. Based on multiple sources of data including students' drafts, automated feedback and submission information, semi-structured interviews and documents, this study explored a cohort of English as a Foreign Language (EFL) students' experiences of appropriating automated feedback in their writing activities. The findings revealed three forms of appropriation (i.e., regular, partial, and rare) among the students, who had used various artifacts (e.g., dictionary) and rules (e.g., teacher requirement), taken on different roles (e.g., student writer, spotlight-avoider), and resorted to different community members (e.g., peers) to mediate their appropriation. The findings also showed that the students further differed in their internalization of the resources through three sub-processes: selecting, emotion-regulating, and goal-setting. These findings pose a critical need to revisit the idea that submission for immediate automated feedback is motivating and to watch the potential washback effect of setting a threshold score when pedagogically using automated feedback. The study argues that students should develop their awareness of the mediating resources and the internationalization strategies for more self-regulated use of automated feedback. [ABSTRACT FROM AUTHOR]

Cardiac hormones act on the regulation of blood pressure (BP) and cardiovascular homeostasis. These hormones include atrial and brain natriuretic peptides (ANP, BNP) and activate natriuretic peptide receptor-A (NPRA), which enhance natriuresis, diuresis, and vasorelaxation. In this study, we established the ANP-dependent homologous downregulation of NPRA using human embryonic kidney-293 (HEK-293) cells expressing recombinant receptor and MA-10 cells harboring native endogenous NPRA. The prolonged pretreatment of cells with ANP caused a time- and dose-dependent decrease in 125I-ANP binding, Guanylyl cyclase (GC) activity of receptor, and intracellular accumulation of cGMP leading to downregulation of NPRA. Treatment with ANP (100 nM) for 12 h led to an 80% decrease in 125I-ANP binding to its receptor, and BNP decreased it by 62%. Neither 100 nM c-ANF (truncated ANF) nor C-type natriuretic peptide (CNP) had any effect. ANP (100 nM) treatment also decreased GC activity by 68% and intracellular accumulation cGMP levels by 45%, while the NPRA antagonist A71915 (1 µM) almost completely blocked ANP-dependent downregulation of NPRA. Treatment with the protein kinase G (PKG) stimulator 8-(4-chlorophenylthio)-cGMP (CPT-cGMP) (1 µM) caused a significant increase in 125I-ANP binding, whereas the PKG inhibitor KT 5823 (1 µM) potentiated the effect of ANP on the downregulation of NPRA. The transfection of miR-128 significantly reduced NPRA protein levels by threefold compared to control cells. These results suggest that ligand-dependent mechanisms play important roles in the downregulation of NPRA in target cells. [ABSTRACT FROM AUTHOR]

Deciphering the endocytosis mechanisms of nanoparticle entry in cells is crucial to understand the fate of nanoparticles and the biological activity of the transported cargo. Such studies require the use of reporter agents such as fluorescent markers. Previously, we have reported the synthesis of self-fluorescent HAp nanoparticles as efficient nucleic acid delivery agents in prokaryotic and eukaryotic cells. Here, we show the application of biocompatible self-fluorescent nano delivery vehicle based on HAp and CPP- octa-arginine as an efficient system to study the mechanisms of intracellular fate of a gene delivery agent. The pathway of octa-arginine functionalized HAp NP (R8HNP) and HAp NP uptake in R1 ESCs was elucidated using confocal microscopy with the help of endocytic inhibitors. The NPs mainly enter R1 ESCs by clathrin mediated and macropinocytosis pathways. It was established that the NPs escape endosomal degradation by proton sponge effect owing to their ability to buffer the pH and trigger osmotic rupture. The functionalization of CPP, effectively enhanced the internalization and endosomal escape in R1 ESCs. The detailed results of these studies are outlined in this manuscript. • Self-fluorescence property of NPs facilitates intracellular tracking in R1 ESCs. • R8HNPs and HAp NPs are internalized in R1 ESCs majorly by endocytosis. • As-synthesized NPs escape endosomal degradation by proton sponge effect. • R8HNPs undergo enhanced escape from endosomes due to presence of CPP- octa-arginine. • Self-fluorescence property of the NPs makes them multifunctional in nature with application in bio-imaging and gene delivery. [ABSTRACT FROM AUTHOR]

Interprofessional collaboration (IPC) in the OR enhances safe and effective patient care. The aim of this qualitative study was to explore perioperative nurses' perspectives on their contributions to IPC. We conducted a secondary analysis of 19 semistructured interviews with perioperative RNs and completed inductive thematic analysis with subsequent categorization of the themes into the Interprofessional Education for Collaborative Patient‐Centred Practice Framework. Nurses expressed the importance of being heard through effective communication, feeling confident in their role, being aware of interdependent roles, and sharing a common understanding. From nurses' perspectives, use of structured processes enabled organization of interdisciplinary patient care. Nurses showed leadership skills when they anticipated the needs of the team and recognized they needed support to develop these skills. They contributed to IPC through their shared understanding of common goals, leadership skills in the OR, and active involvement in delivering structured processes. [ABSTRACT FROM AUTHOR]

Synaptic strength is altered during synaptic plasticity by controlling the number of AMPA receptors (AMPARs) at excitatory synapses. During long-term potentiation and synaptic upscaling, AMPARs are accumulated at synapses to increase synaptic strength. Neuronal activity leads to phosphorylation of AMPAR subunit GluA1 (also known as GRIA1) and subsequent elevation of GluA1 surface expression, either by an increase in receptor forward trafficking to the synaptic membrane or a decrease in receptor internalization. However, the molecular pathways underlying GluA1 phosphorylation-induced elevation of surface AMPAR expression are not completely understood. Here, we employ fluorescence recovery after photobleaching (FRAP) to reveal that phosphorylation of GluA1 serine 845 (S845) predominantly plays a role in receptor internalization, rather than forward trafficking, during synaptic plasticity. Notably, internalization of AMPARs depends upon the clathrin adaptor AP2, which recruits cargo proteins into endocytic clathrin-coated pits. In fact, we further reveal that an increase in GluA1 S845 phosphorylation upon two distinct forms of synaptic plasticity diminishes the binding of the AP2 adaptor, reducing internalization and resulting in elevation of GluA1 surface expression. We thus demonstrate a mechanism of GluA1 phosphorylation-regulated clathrin-mediated internalization of AMPARs. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus main internalization mechanism in osteoblasts relies on a tripartite interaction between bacterial fibronectin-binding proteins, extracellular matrix soluble fibronectin, and osteoblasts' β1 integrins. Caveolins, and particularly caveolin-1, have been shown to limit the plasma membrane microdomain mobility, and consequently reduce the uptake of S. aureus in keratinocytes. In this study, we aimed to deepen our understanding of the molecular mechanisms underlying S. aureus internalization in osteoblasts. Mechanistically, S. aureus internalization requires endosomal recycling of β1 integrins as well as downstream effectors such as Src, Rac1, and PAK1. Surprisingly, in β1 integrin deficient osteoblasts, S. aureus internalization is restored when Caveolin-1 is absent and requires αvβ3/5 integrins as backup fibronectin receptors. Altogether, our data support that β1 integrins regulate the level of detergent-resistant membrane at the plasma membrane in a an endosomal and Caveolin-1 dependent manner., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Weight stigma (devaluation due to body weight) in healthcare is common and influences one's engagement in healthcare, health behaviors, and relationship with providers. Positive patient-provider relationships (PPR) are important for one's healthcare engagement and long-term health., Purpose: To date, no research has yet investigated whether weight bias internalization (self-stigma due to weight; WBI) moderates the effect of weight stigma on the PPR. We predict that weight stigma in healthcare is negatively associated with (i) trust in physicians, (ii) physician empathy, (iii) autonomy and competence when interacting with physicians, and (iv) perceived physician expertise. We also predict that those with high levels of WBI would have the strongest relationship between experiences of weight stigma and PPR outcomes., Methods: We recruited women (N = 1,114) to complete a survey about weight stigma in healthcare, WBI and the previously cited PPR outcomes., Results: Weight stigma in healthcare and WBI were associated with each of the PPR outcomes when controlling for age, BMI, education, income, race, and ethnicity. The only exception was that WBI was not associated with trust in physicians. The hypothesis that WBI would moderate the effect of weight stigma in healthcare on PPR outcomes was generally not supported., Conclusions: Overall, this research highlights how weight stigma in healthcare as well as one's own internalization negatively impact PPRs, especially how autonomous and competent one feels with their provider which are essential for one to take an active role in their health and healthcare., (© Society of Behavioral Medicine 2024. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Wiskott-Aldrich syndrome (WAS) is an X-linked immunodeficiency condition caused by ablation of functional WAS protein (WASP) expression, and associated with susceptibility to infections, eczema, and autoimmunity. Regulatory T cell (Treg) defects are an important cause of autoimmunity in WAS. Currently, the mechanisms underlying cytoskeleton involvement in Treg-regulated autoimmunity remain unclear, and WAS is an excellent model for investigation of this question. Here, we examined patients with WAS and WASP knockout (WASp -/- ) mice to uncover a new mechanism involving the actin nucleation promoting factor, WASP, in regulating Treg tolerance by modulating their surface IL-2 receptor (IL-2R) levels. Surface expression levels of IL-2R and its downstream signaling molecules, phosphoinositide 3-kinase/pSTAT5, are decreased in WASp -/- Tregs. Low dosage IL-2 combined with anti-IL-2 monoclonal antibody (IL2 complex) treatment can compensate for Treg deficiency in WAS in vitro and in vivo. Moreover, IL2 complex treatment relieved autoimmune colitis in WASp -/- mice. Reduced surface IL-2R is primarily caused by elevated IL-2R internalization and degradation, and lysosomal and endosomal genes associated with these processes are upregulated in WASp -/- Tregs. Finally, spatiotemporal analysis of dynamin and Neural Wiskott Aldrich Syndrome Protein (N-WASP) recruitment, by generating lipid bilayers and total internal reflection fluorescence microscopy, showed that WASP deficiency promoted IL-2R internalization and degradation by enhancing N-WASP activation. Consistently, N-WASP inhibition in Tregs using wiskostatin reduced IL-2R internalization. Together, our results reveal a novel intrinsic role of WASP in regulation of surface IL-2R dynamics in Tregs, highlighting a potential new therapeutic approach for autoimmune diseases., Competing Interests: Declaration of competing interest None., (Copyright © 2024. Published by Elsevier Ltd.)

The Zika virus (ZIKV) has garnered significant public attention, particularly following the outbreak in Brazil, due to its potential to cause severe damage to the central nervous system and its ability to cross the placental barrier, resulting in microcephaly in infants. Despite the urgency, there remains a lack of targeted therapies or vaccines for the prevention or treatment of ZIKV infection and its related diseases. Fangchinoline (FAN), an alkaloid derived from traditional Chinese medicinal herbs, has a range of biological activities. In this study, we employed both in vitro and in vivo infection models to demonstrate the efficacy of FAN in inhibiting ZIKV. Our findings indicate that FAN effectively suppresses the replication of ZIKV viral RNA and protein, thereby validating its anti-ZIKV capabilities in living organisms. Further analysis through dosing time assays and infectious inhibition assays revealed that FAN exerts its antiviral effects by impeding the early stages of infection, specifically by inhibiting the internalization of ZIKV. These results underscore the potential of FAN as a candidate for anti-ZIKV drug development and offer novel insights into drug design strategies that target the virus's internalization process.

The sarco (endo)plasmic reticulum Ca 2+ -ATPase 2a (SERCA2a)-phospholamban (PLN) system within the sarcoplasmic reticulum is crucial for regulating intracellular Ca 2+ cycling in ventricular cardiomyocytes. Given that impaired Ca 2+ cycling is associated with heart failure, modulating SERCA2a activity represents a promising therapeutic strategy. Previously, we engineered an RNA aptamer (Apt30) that binds to PLN, thereby activating SERCA2a by alleviating PLN's inhibitory effect. However, Apt30 alone cannot reach intracellular PLN, necessitating the development of a mechanism for its specific internalization into cardiomyocytes. Using the systematic evolution of ligands by exponential enrichment (SELEX) method, we isolated RNA aptamers capable of internalizing into cardiomyocytes. These aptamers demonstrated sub-micromolar EC 50 values for cardiomyocyte internalization and exhibited significantly reduced activity against various non-myocardial cells, highlighting their specificity for cardiomyocytes. Moreover, some of these cardiomyocyte-internalizing aptamers could be linked to Apt30 as a single RNA strand without compromising their internalization efficacy. Supplementing the culture medium with these hybrid aptamers enhanced Ca 2+ transients and contractile function in rat cardiomyocytes. These findings provide critical insights for developing novel therapeutics directly acting on PLN in cardiomyocytes, potentially compensating for the disadvantages of conventional methods that involve viral vector-mediated intracellular transduction or alterations in endogenous protein expression., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Arrestins are essential proteins for the regulation of G protein-coupled receptors (GPCRs). They mediate GPCR desensitization after the activated receptor has been phosphorylated by G protein receptor kinases (GRKs). In addition, GPCR-arrestin interactions may trigger signaling pathways that are distinct and independent from G proteins. The non-visual GPCRs encompass hundreds of receptors with varying phosphorylation patterns and amino acid sequences, which are regulated by only two human non-visual arrestin isoforms. This review describes recent findings on GPCR-arrestin complexes, obtained by structural techniques, biophysical, biochemical, and cellular assays. The solved structures of complete GPCR-arrestin complexes are of limited resolution ranging from 3.2 to 4.7 Å and reveal a high variability in the relative receptor-arrestin orientation. In contrast, biophysical and functional data indicate that arrestin recruitment, activation and GPCR-arrestin complex stability depend on the receptor phosphosite sequence patterns and density. At present, there is still a manifest lack of high-resolution structural and dynamical information on the interactions of native GPCRs with both GRKs and arrestins, which could provide a detailed molecular understanding of the genesis of receptor phosphorylation patterns and the specificity GPCR-arrestin interactions. Such insights seem crucial for progress in the rational design of advanced, arrestin-specific therapeutics., (© 2024 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)