Interaction Sites Research Articles

Morpholine-tethered Novel Hydrazones as Promising Non-peptidic Prolyl Oligopeptidase (POP) Inhibitors: Synthesis In Vitro and In Silico Studies.

Prolyl oligopeptidase (POP) is a pivotal druggable target implicated in diverse biological processes and linked to the development of various ailments, including neurodegenerative disorders. While conventional peptide-based inhibitors have been a centerpiece, their limitations, such as restricted bioavailability, necessitate exploration of non-peptidic inhibitors for their therapeutic potential. This study focuses on designing, synthesizing, and assessing morpholine-based hydrazones targeting the catalytic serine residue of POP. The hydrazones (5a-o), reported as moderately potent analogs compared to the renowned Z-Pro-Prolinal, demonstrated in vitro POP inhibition with IC50 values ranging from 13.60 ± 2.51 to 36.51 ± 1.82 μM. The derivative 5h, with an IC50 of 13.60 ± 2.51 μM, emerged as the most potent inhibitor. Moreover, the in vitro kinetic study of compound 5h indicated that it exhibited concentration-dependent type of inhibition. in silico docking studies of 5h revealed robust interactions in the POP enzyme's active site, yielding a docking score of -6.30 Kcal/- mol, consistent with experimental results. All findings underscored the potential of synthesized derivatives for drug development.

Education with interactif website media on patient compliance levels for implementation of post-odontectomy instructions

Introduction: Patient compliance with post odontectomy instructions is one of the factors that determine postoperative success. Lack of patient knowledge can interfere with the healing process and cause complications. Instructions given verbally often make patients less aware of post-operative care. Interactive website media through gadgets can be used as a tool to increase patient knowledge as an effort to prevent complications. The purpose of the study was to determine the effect of providing education with interactive website media on the level of patient compliance with post-odontectomy instructions. Methods: The type of research used is pre-experiment research with Post Test Only With Control Group Design. Sampling using Accidental Sampling technique obtained as many as 30 people. The variables in the study were interactive website media and compliance. Hypothesis testing using mann whitney U. Results: The results showed a difference in the level of compliance after being given education through interactive website media between the intervention group (80%) and the control group (53.35%). Mann Whitney U analysis showed that there was an effect on the provision of education with interactive website media on the level of patient compliance with the implementation of post odontectomy instructions p value = 0.049 (p < 0.05). Conclusion Interactive website media can improve patient compliance with instructions given at post odontectomy. Odontectomy patients become more aware of the importance of carrying out postoperative instructions to prevent complications

Electrically conductive polyaniline: Graphite composite on porous polyurethane sponge with enhanced sensitivity of humidity

AbstractRecent advancements in organic materials based sensor technology created a prospective field in designing sensors that are both cost‐effective and environment friendly. In view of that an electronic material system has been prepared depositing a uniform layer of graphite‐infused Polyaniline (PANI) onto the sponge framework using a straightforward physical vapor deposition technique. The prepared active materials were studied for the composition, morphology, optical properties, and charge transport characteristics. In the current context of rapidly growing sensor technology, this works describes an innovative approach of improving the response of the prepared electronic system of PANI/Gr composites, integrated into porous PU sponge substrates. Incorporation of graphite in PANI improved the electrical conductivity of the composite and porous structure of PU increased the interaction surface area. The performance of the prepared materials in humidity detection was evaluated by studying their resistive response under varying humidity levels, which was measured through current‐voltage (I‐V) characteristics. The higher interaction sites of the reported active sensing system leads to inspiring humidity sensitivity of 0.522–26.22 kΩ/% RH with reasonable response and recovery time of 572 and 416 sec respectively. Additionally, the reported sensor system consisting of degradable materials will offer a useful way of reducing electronic garbage.

WDR64, a testis-specific protein, is involved in the manchette and flagellum formation by interacting with ODF1

The WD40 repeat (WDR) domain is present in a wide range of proteins, providing sites for protein‒protein interactions. Recent studies have shown that WDR proteins play indispensable roles in spermatogenesis, such as in spermatocyte division, sperm head formation and flagellar assembly. In this study, we identified a novel testis-specific gene, WDR64, which has the typical characteristics of WD40 proteins with two β-propellers, and is highly conserved in Mammalia. RT-PCR and Western blot results revealed that WDR64 was highly expressed in testis. WDR64 protein was weakly expressed at postnatal Day 7, increased substantially at postnatal Day 28 and maintained at high levels thereafter. Further immunofluorescence demonstrated that WDR64 was localized posterior to the nucleus in steps 8–14 spermatids in line with the dynamic localization of manchette, moved to the flagella in steps 15–16 spermatids, and localized at the midpiece of the flagellum in mature spermatozoa. To explore the function of WDR64, we performed immunoprecipitation‒mass spectrometry (IP‒MS) to screen its interacting proteins and found that WDR64 interacted with ODF1 to form a complex. The WDR64/ODF1 complex is located at the manchette during nucleus shaping and finally at the midpiece of the mature spermatozoa tail, suggesting that it may be involved in the assembly of the manchette and flagella during spermiogenesis. Our findings provide the first understanding of the expression pattern of WDR64 and its potential molecular mechanism in spermiogenesis.

Prospects and challenges for parliamentarism in the conditions of the development of digital constitutionalism

The purpose of the article is to carry out a multi-faceted analysis of the dynamics of the development of parliamentarism and digital constitutionalism, to identify possible challenges and threats, as well as prospects for parliaments and parliamentary activity in the conditions of the spread of modern digital (electronic) technologies. Approaches to understanding and defining the phenomena of «parliamentarism», «constitutionalism», «digital constitutionalism», some of their modifications and features are highlighted, their multifaceted nature, transformation in the conditions of modern progressive digital development, electronic democracy is emphasized. Crisis and conflict events have largely become challenges for parliaments and parliamentary activity, as well as digitalization and digitization processes and tools. It is important that parliamentary institutions of various levels (national, supranational, inter-parliamentary associations, their auxiliary bodies and services) contribute to the development and regulation of these processes, using advanced and effective digital technologies tested in practice. The «Digital Compass» for the EU Digital Decade (2030), the concept of digital sovereignty, the adopted charters of digital rights (Catalan, Nice), the Law on Digital Markets, the Law on Digital Services, the Law on Artificial Intelligence, the Declaration on European Digital Rights and Principles should be singled out. (2022), «World e-Parliament Report» of the IPU, the concept of e-democracy of the European Parliament, the resolution on e-democracy of the PACE, recommendations on open government and electronic participation of citizens OECD. Thanks to this, new forms and tools for the implementation of existing human rights are introduced, as well as new digital rights are constituted (to access the Internet, to identity in the digital environment, to protection in the digital environment, non-discrimination, etc.). In the same way, the forms and methods of activity of parliaments are being modernized (expansion of platforms for online broadcasts of plenary and committee meetings, electronic (remote) voting, improvement of interactive websites with access to draft laws and other documents, and systems of electronic petitions and consultations with voters, electronic participation of citizens and public organizations in the legislative process). However, the development of digital parliamentarism and digital democracy is accompanied by numerous challenges (digital inequality, low digital literacy of members of parliaments and parliamentary services, cyber threats, material and financial costs etc.). This requires a thorough study of the issues raised, improvement of digital culture, improvement of parliamentary and non-parliamentary procedures, along with further development of technological infrastructure, cyber security protection of the components of digital parliamentarism.

Silencing EPHB2 diminished the malignant biological properties of esophagus cancer cells by blocking autophagy and Wnt/β-catenin pathway.

Eph receptor B2 (EPHB2) is overexpressed in some tumors and relevant to unfavorable outcomes of tumor patients. By searching Gene Expression Profiling Interactive Analysis and KM Plot websites, we discovered that EPHB2 was highly expressed in patients with esophageal cancer, leading to poor prognosis. However, the role and molecular mechanism of EPHB2 in esophagus cancer is unknown. Our study aims to unveil the underlying mechanism by which EPHB2 modulates the biological properties of esophagus cancer cells. After si-EPHB2 transfection, the malignant biological properties of esophagus cancer cells were determined by several biological experiments. IWP-4 was applied to block Wnt/β-catenin signaling pathway. The expressions of autophagy and Wnt/β-catenin signaling pathway relevant molecules were tested by western blot assay. An increased expression of EPHB2 was happened in esophagus cancer samples and loss of EPHB2 diminished esophagus cancer cells proliferation, migration, and invasion. Moreover, our data showed that depletion of EPHB2 blocked the autophagy and in-activated Wnt/β-catenin signaling pathway in esophagus cancer cells. While, IWP-4 treatment inhibited the autophagy and limited esophagus cancer cells proliferation, migration, and invasion. Moreover, EPHB2 knocked down strengthened the effect of IWP-4 treatment in regulating esophagus cancer cells proliferation, migration, and invasion. Finally, we illustrated that EPHB2 regulated the biological properties of esophagus cancer cells by modulating autophagy and Wnt/β-catenin signaling pathway. Our study illustrated that EPHB2 might be a worthwhile target considering for the treatment of esophagus cancer.

Comparative Analysis of Inhibitory and Activating Immune Checkpoints PD-1, PD-L1, CD28, and CD86 in Non-Melanoma Skin Cancer.

The establishment of immunotherapy applying immune checkpoint inhibitors (ICI) has provided an important new option for the treatment of solid malignant diseases. However, different tumor entities show dramatically different responses to this therapy. BCC responds worse to anti-PD-1 ICIs as compared to cSCC. Differential immune checkpoint expression could explain this discrepancy and, therefore, the aim of this study was to analyze activating and inhibitory immune checkpoints in cSCC and BCC tissues. Tissue microarrays of the invasive front as well as the tumor core of BCC and cSCC samples were used to evaluate PD-1, PD-L1, CD28, and CD86 expression and their topographic distribution profiles by chromogenic immunohistochemistry. QuPath was used to determine the labeling index. The expression of PD-1, PD-L1, and CD28 was significantly higher in both the tumor core and the invasive front of cSCC samples as compared to BCC (p < 0.001). In addition, the ratios of PD-L1/CD86 (p < 0.001) and CD28/CD86 (p < 0.001) were significantly higher in cSCC. The invasive front of both tumor entities showed higher expression levels of all immune markers compared to the tumor core in both tumor entities. The significantly higher expression of PD-1, PD-L1, and CD28 in cSCC, along with the predominance of the inhibitory ligand PD-L1 as compared to the activating CD86 in cSCC, provide a potential explanation for the better objective response rates to anti-PD-1 immunotherapy as compared to BCC. Furthermore, the predominant site of interaction between the immune system and the tumor was within the invasive front in both tumor types.

Structural basis of CDNF interaction with the UPR regulator GRP78

Cerebral dopamine neurotrophic factor (CDNF) is an unconventional neurotrophic factor that is a disease-modifying drug candidate for Parkinson’s disease. CDNF has pleiotropic protective effects on stressed cells, but its mechanism of action remains incompletely understood. Here, we use state-of-the-art advanced structural techniques to resolve the structural basis of CDNF interaction with GRP78, the master regulator of the unfolded protein response (UPR) pathway. Subsequent binding studies confirm the obtained structural model of the complex, eventually revealing the interaction site of CDNF and GRP78. Finally, mutating the key residues of CDNF mediating its interaction with GRP78 not only results in impaired binding of CDNF but also abolishes the neuroprotective activity of CDNF-derived peptides in mesencephalic neuron cultures. These results suggest that the molecular interaction with GRP78 mediates the neuroprotective actions of CDNF and provide a structural basis for development of next generation CDNF-based therapeutic compounds against neurodegenerative diseases.

Deciphering the adsorption machinery of Deep-Blue and Vp4, two myophages targeting members of the Bacillus cereus group.

The Bacillus cereus group comprises closely related species, including some with pathogenic potential (e.g., Bacillus anthracis and Bacillus cytotoxicus). Their toxins represent the most frequently reported cause of food poisoning outbreaks at the European level. Bacteriophage research is undergoing a remarkable renaissance for its potential in the biocontrol and detection of such pathogens. As the primary site of phage-bacteria interactions and a prerequisite for successful phage infection, adsorption is a crucial process that needs further investigation. The current knowledge about B. cereus phage adsorption is currently limited to siphoviruses and tectiviruses. Here, we present the first insights into the adsorption process of Herelleviridae Vp4 and Deep-Blue myophages preying on B. cereus hosts, highlighting the importance of polysaccharide moieties in this process and confirming the binding to the host surface of Deep-Blue Gp185 and Vp4 Gp112 receptor-binding proteins and Gp119 baseplate wedge.

Exploring Chemical Space to Identify Partial Binders Against hMPV Nucleocapsid Protein.

Human metapneumovirus (hMPV) has gained prominence in recent times as the predominant etiological agent of acute respiratory tract infections. This virus targets children, the elderly, and individuals with compromised immune systems. Given the protracted duration of hMPV transmission, it is probable that the majority of children will have acquired the virus by the age of 5. In individuals with compromised immune systems, recurrence of hMPV infection is possible. As hMPV matures, it remains latent from the time of acquisition. The genome of hMPV encompasses a pivotal protein referred to as the nucleocapsid protein (N). This protein assumes the form of a left-handed helical nucleocapsid, enveloping the viral RNA genome. The primary function of this structure is to protect nucleases, rendering it a potentially promising target for therapeutic advancements. The present study employs a methodology that involves structure-based virtual screening, followed by molecular dynamics simulation at a 250-ns time scale, to identify potential natural molecules or their derivatives from the ZINC Database. These molecules are investigated for their binding properties against the hMPV nucleoprotein. Based on an evaluation of the docking score, binding site interaction, and molecular dynamics studies, it has been found that two naturally occurring molecules, namely M1 (ZINC85629735) and M3 (ZINC85569125), have shown notable docking scores of -9.6and -10.7 kcal/mol, acceptable RMSD, RMSF, Rg, and soon calculated from molecular dynamics trajectory associated with MMGBSA binding energy of -81.94 and -99.63 kcal/mol, respectively. These molecules have shown the highest binding affinity towardnucleocapsid protein and demonstrated promising attributes as potential binders against hMPV.

Women of Gülhane: multiple femininities and gendered spaces of ‘The Festival of Spring and Flowers’

This paper explores the discursive and spatial construction of feminine norms during the ‘Festival of Spring and Flowers,’ a momentous urban event held at Gülhane Park, İstanbul, from 1950 to 1960. Gülhane was a bustling and highly frequented location during the Festival, serving as a significant site of convergence, interaction, and contestation. Consequently, Gülhane provides a crystallized and concise cross-section of the ubiquitous femininities in 1950s Istanbul. Drawing upon textual and visual archival evidence, the author(s) argue that the Festival played a pivotal role in perpetuating gendered norms by consistently portraying ‘liberated, domesticated, and pariah’ femininities. Concomitant with the trifold construction of femininity, the space itself took on a ‘gendered persona’; Gülhane was depicted as a safe and sterile location when frequented by the ‘liberated and domesticated’ women, but was perceived as an ‘immoral’ space when ‘invaded’ by the ‘pariah’ women. The gendered history of 1950s İstanbul have been largely overlooked in the current scholarship, despite the emergence of new gendered norms alongside the transforming political landscape in Turkey during that time. This article will address this literature gap by conducting an in-depth historical analysis of the gender norms within the Festival. Additionally, this study will contribute to İstanbul’s urban literature by exploring the notable yet relatively uncharted Festival.

Stereoisomeric separation and chiral recognition mechanism study of star cyclodextrin polymer as the chiral stationary phase

BackgroundAs the derivatives of cyclodextrin (CD), cyclodextrin polymers (CDPs) effectively increase the concentration of CD units and construct supramolecular structures with unique stereoselectivity by the structure design. CDPs have shown significant potential in chiral separation, however, the process of stereoselective interactions on chiral stationary phases (CSPs) and the specific contribution of intermolecular forces are still a challenge issue. A comprehensive understanding of the chiral recognition mechanism of CDPs will help to optimize chiral separation conditions and design new CSPs. ResultsThe star CDP with a supermolecular structure was synthesized by grafting β-CD onto the external 6-position hydroxyl groups using β-CD as the parent nucleus. The enhanced host-guest recognition ability of CD supramolecular polymer structure provided better inclusion interaction and increased chiral recognition of the isomers. The Star-CD CSP with star CDP as a chiral ligand performed satisfactory stereoisomer separation ability with the separation factor (α) up to 2.0 for various quinoline alkaloid isomers and 1.89 for catechins. To elucidate its chiral separation mechanism, molecular docking was used to construct the three-dimensional visual models of the binding sites and the contribution of non-covalent interactions between Star-CD CSP and quinoline alkaloid isomers. In addition, the formation sites of non-covalent interactions on the CD monomers of the polymer side chains were confirmed from the actual geometric structure by analyzing the NMR chemical shift changes before and after the formation of complexes between Star-CD polymers and isomers. Combined with the mutual evidence of molecular simulation and chiral NMR, the specific recognition mechanism of selector-selectand complexes was comprehensively expounded. SignificanceThe multi-mode CSP based on cyclodextrin supramolecular structure provides new ideas for the stereoisomeric separation of complex chiral components with multiple chiral centers in natural products. Not limited to the macroscopic performance of the chromatographic separation, molecular docking explored the theoretical model of chiral recognition from the molecular level. The chiral NMR analysis confirmed the credibility of the model from the geometry structure, and then the recognition mechanism of multi-mode CSP was fully elaborated combining the above three aspects.

Coupling of the catalytic reactions from formate dehydrogenase and hydrogenase in solution: Insights from computations and in situ IR spectroscopy

Sophisticated enzymatic systems have evolved in nature to efficiently couple distinct biochemical reactions in form of cascades. As such they serve as reference models to understand the indirect interactions of catalytic centers. Herein, we studied, in solution, the coupling of the reactions from membrane‐bound [NiFe] hydrogenase (MBH) from Cupriavidus necator (reversible H2 splitting into H+ and e‐) and the molybdenum‐dependent formate dehydrogenase from Rhodobacter capsulatus (reversible formate to CO2 interconversion). To follow their interplay via the characteristic absorptions from the MBH's active site or the respective substrate and product bands of FDH, we utilized in situ IR spectrocopy and GC(‐MS), in the absence or presence of soluble redox mediators. Coarse grained molecular dynamics (cgMD) computations revealed the lack of productive enzyme complexes for direct electron transfer (ET). Thus, the observed minor amounts of H2 or formate were produced from transient interactions between the two enzymes. On the contrary, the significantly increased product formation in the presence of methylene viologen can be related to the putative multiple interaction sites of the redox mediator with FDH identified by cgMD. Our study represents a proof‐of‐concept approach that can be used in future to develop novel coupled biocatalytic systems by identifying potential ET pathways.

Analysis of the sweetening effects of lactone and ketone compounds in a cheddar cheese matrix using sensory analysis and molecular simulation

This study evaluated the effects of lactone and ketone compounds on sweetness perception in a Cheddar cheese matrix. According to the static sensory evaluation, a ternary mixture of γ-decanolactone/δ-decanolactone/acetoin at middle and high concentrations exhibited a significant sweetening effect closely related to olfactory input. Dynamic sensory evaluation showed that a γ-decanolactone/δ-decanolactone/acetoin ternary mixture at middle and high concentrations could enhance the sweetness perception of a Cheddar cheese matrix at different sucrose concentrations during oral processing. Molecular docking showed that the ternary mixture of γ-decanolactone/δ-decanolactone/acetoin with sucrose-T1R2 increased the number of hydrogen bond binding sites and increased the number of hydrophobic interaction sites. Molecular dynamics suggested that the molecular conformation of sucrose and T1R2 was more compact and stable in the ternary mixture. Additionally, in the presence of the ternary system, the binding free energy of the re-docking of sucrose and T1R2 was −7.28 kcal/mol, indicating that the docking structure is stable. This stability is maintained by the hydrogen bonding and hydrophobic interaction between the T1R2-sucrose complex and aroma mixtures. These findings provide a theoretical basis for using cross-modal perception to sweeten Cheddar cheese to cater to consumers’ pursuit of sweetness.

Identification of the Potential Role of the E4orf4 Protein in Adenovirus A, B, C, and D Groups in Cancer Therapy: Computational Approaches.

The human adenovirus (HADV) early region 4 open reading frame 4 (E4orf4) protein plays a regulatory role in promoting viral infection by interacting with various cellular proteins. E4orf4 can induce death in cancer cells. One of the death pathways that is induced by this protein is related to the formation of membrane blebbing following the phosphorylation of tyrosine amino acids. The activation of this pathway requires the interaction of E4orf4 with Src family kinases (SFKs). The modulation mechanism of Src-dependent signaling via E4orf4 is not yet fully understood. However, evidence suggests that a physical association between the Src kinase domain and the arginine-rich motif of E4orf4 is crucial. Physically connecting E4orf4 to Src kinase leads to the deregulation of the Src-related signaling pathway, thereby inducing cytoplasmic death. In this study, we mapped the E4orf4 interaction site in Src to investigate the interaction between E4orf4 and Src in detail. We also compared the binding strength of E4orf4 proteins from different HADV groups. To this end, we performed bioinformatics structural analysis of the Src kinase domain and E4orf4 to identify E4orf4 interaction sites. The group with the lowest binding energy was predicted to be the most likely candidate for the highest cytoplasmic death in tumor cells based on the energy of the E4orf4-Src complex in various HADV groups. These results show that HADV-A and HADV-C have minimal binding energies to the E4orf4-Src complex, while the dissociation constant (Kd) of HADV-A was less than that of HADV-C. According to the obtained results, E4orf4 of the HADV-A group is more effective at triggering cytoplasmic death based on its most robust interaction with the Src kinase domain.

A combined multilevel computational and experimental approach for interfacial screening and mechanical reinforcement of basalt fiber/polyamide 66 composites

Silane coupling agents (SCAs) can improve fiber-resin interface issues, yet selecting an appropriate SCA experimentally is resource-intensive. Herein, a multi-level screening method combining simulation and experiments is proposed to identify SCA-enhancing bonding between SCAs and polyamide 66 (PA66) and the basalt fiber (BF) −PA66 interface. SCAs with organic groups composed of N and O exhibited superior interaction with PA66 due to the more pronounced negative and positive electrostatic potential (ESP) distributions. In SCAs-grafted BF-PA66 models, SCAs not only altered BF surface nanostructure but also changed the arrangement of PA66 main chains from parallel to interpenetrating BF surface atoms. Besides, (3-ureapropyl)trimethoxysilane (SCA6) significantly improved BF-PA66 bonding, which was experimentally validated by enhanced interlaminar shear and mechanical properties. O atoms on SCA6 and PA66 are more electronically bound and have the smallest ESP extremum, providing strong polar interaction sites. Additionally, both simulations and experiments demonstrated a transformation from adhesive to cohesive failure.

Sulfonated polybenzimidazole membrane with graphene oxide additive for 2,3-butanediol/water separation: A molecular simulation

Membrane separation for 2,3-butanediol (2,3-BDO) recovery from fermentation broth is highly valued for sustainable and renewable processes, but it requires efficient membrane materials. This work evaluates the sulfonated polybenzimidazole (sPBI) and its graphene oxide (GO) doped composite membrane for separating 2,3-BDO and water via atomistic simulations. Density functional theory calculations are applied to identify various forms of sPBI structures and quantify their binding interactions with 2,3-BDO and water. Classical molecular dynamic simulations are used to evaluate the structural changes, diffusivity, and selectivity of 2,3-BDO and water in different sPBI models, GO surfaces, and GO-doped sPBI composite models. Our results suggest that sPBI slightly increases the crystallinity of the membrane structures, enhances the adsorption strength for both 2,3-BDO and water, and improves the water/2,3-BDO selectivity by 2–3 times. The GO surfaces display a maximum selectivity at a surface coverage of 0.1–0.15 for both hydroxyl and epoxy surface groups. The addition of GO flakes to sPBI creates new interaction sites for 2,3-BDO and water at the interface of sPBI and GO, and the water/2,3-BDO selectivity of GO-doped sPBI models is further increased up to 3 times. This work illustrates how the integrated addition of sPBI and GO flakes offers a promising approach to selective separation of 2,3-BDO and water, providing theoretical guidance for polybenzimidazole-based membranes in the potential application of 2,3-BDO recovery.

The hybrid authenticity of virtual pilgrimage

The complex relationship between religion and markets deserves nuanced scholarly reflection and observation. The continuing explosion of online or digital expressions of religion invites researchers to consider the authenticity of virtual pilgrimage experience, for which it is situated at the intersections between religious and marketing studies that allows various dimensions of authenticity to take place. The key contribution of this conceptual paper lies in its development and application of a ‘Hybrid authenticity’ framework which helps explain the consumer-congregants’ development of perceived authenticity of virtual pilgrimage. This framework invites a shift in focus towards the prioritization of iconic-authenticity cues that facilitates a much more ‘liquid’ attachment to object-based authenticity. It also emphasizes the importance of place as an element of constructive authenticity in interpreting pilgrims’ experience and the vital contribution of integrating both interpersonal and intrapersonal authenticity. We suggest the mechanism of this hybrid framework is defined by the interdependence between its elements. We also discuss the two underlying conditions for the mechanism; technology working as an embodied site of interaction and consumer (re)negotiation of authenticity. Viewing virtual pilgrimage as part of a broader social and cultural transformation, we propose the framework also has applicability to other marketing and post-postmodern consumption studies.

SARS-CoV-2 envelope protein alters calcium signaling via SERCA interactions

The clinical management of severe COVID-19 cases is not yet well resolved. Therefore, it is important to identify and characterize cell signaling pathways involved in virus pathogenesis that can be targeted therapeutically. Envelope (E) protein is a structural protein of the virus, which is known to be highly expressed in the infected host cell and is a key virulence factor; however, its role is poorly characterized. The E protein is a single-pass transmembrane protein that can assemble into a pentamer forming a viroporin, perturbing Ca2+ homeostasis. Because it is structurally similar to regulins such as, for example, phospholamban, that regulate the sarco/endoplasmic reticulum calcium ATPases (SERCA), we investigated whether the SARS-CoV-2 E protein affects the SERCA system as an exoregulin. Using FRET experiments we demonstrate that E protein can form oligomers with regulins, and thus can alter the monomer/multimer regulin ratio and consequently influence their interactions with SERCAs. We also confirm that a direct interaction between E protein and SERCA2b results in a decrease in SERCA-mediated ER Ca2+ reload. Structural modeling of the complexes indicates an overlapping interaction site for E protein and endogenous regulins. Our results reveal novel links in the host-virus interaction network that play an important role in viral pathogenesis and may provide a new therapeutic target for managing severe inflammatory responses induced by SARS-CoV-2.

Intermolecular disulfide bond of PRRSV GP5 and M facilitates VLPs secretion and cell binding

Porcine reproductive and respiratory syndrome virus (PRRSV), the causative agent of porcine reproductive and respiratory syndrome (PRRS), continues to significantly impact on the global swine industry. GP5 and M are the primary structural proteins of PRRSV, playing crucial roles in the processes of virus attachment, entry, assembly and budding. The co-expression of GP5 and M can result in the formation of virus-like particles (VLPs). However, the underlying mechanisms remain incompletely understood. This study investigated the role of GP5-M interaction in VLPs secretion and cell binding. VLPs were generated by co-expressing GP5 and M via recombinant baculoviruses in Sf9 cells and confirmed by transmission electron microscopy. The secretion of VLPs was modulated by the expression levels of GP5 and M. Using the BirA technique, the GP5-M interaction was confirmed in Sf9 cells. Disruption of the N-terminally intermolecular disulfide bond between GP5 and M weakened, but did not completely abolish, the interaction between the proteins, leading to reduced VLPs secretion. Notably, the absence of this intermolecular disulfide bond resulted in the loss of VLPs’ ability to bind to MARC-145 cells. In summary, our findings reveal the critical function of the intermolecular disulfide bond in GP5-M interaction, which significantly contributes to VLPs secretion and cell binding, and suggest potential interaction sites between GP5 and M.