Integrin αVß3 mediates porcine deltacoronavirus infection and inflammatory response through activation of the FAK-PI3K-AKT-nf-κB signalling pathway.

Porcine deltacoronavirus (PDCoV) is an emerging porcine enteropathogenic coronavirus that causes acute watery diarrhoea in piglets, resulting in significant economic losses to the global swine industry. However, the underlying mechanism of PDCoV infection is not well defined, which seriously hinders the development of effective drugs and vaccines. Integrins (ITG) are heterodimeric transmembrane glycoproteins that play important roles in the life cycle of many viruses. In the current study, the viral entry pathways of PDCoV were explored and the role of ITGαVß3 was investigated during PDCoV infection. Our results showed that the lysosomal acidification inhibitor bafilomycin-A1 (Baf-A1) significantly reduced PDCoV infection, while exogenous protease facilitated PDCoV infection and even allowed PDCoV entry to bypass the endosomal pathway, suggesting PDCoV entry into cells via the endocytic pathway and the exogenous protease-mediated pathway simultaneously. Furthermore, ITGαVß3 was identified to be involved in PDCoV infection, especially during viral entry stages. PDCoV infection triggers the activation of the focal adhesion kinase (FAK)-phosphatidylinositol 3-kinase (PI3K)-serine/threonine-specific protein kinase (AKT) signalling pathway, and this activation is ITGαVß3-dependent, suggesting that the activation of the FAK-PI3K-AKT signalling pathway during PDCoV infection is mediated by ITGαVß3. Our results further demonstrated that PDCoV infection induced the expression of inflammatory cytokines, which was mediated by activation of the ITGαVß3-FAK-PI3K-AKT-nuclear transcription factor-κB (NF-κB) signalling pathway. Overall, the results revealed that ITGαVß3 is an essential host factor for PDCoV infection and can serve as a supplementary receptor to facilitate PDCoV infection, which can help us to explore the molecular mechanism of PDCoV infection.

Identifying the host factors required for entry will be helpful in uncovering the pathogenesis mechanisms and developing antivirals against the emerging coronavirus porcine deltacoronavirus (PDCoV). Herein, we revealed that PDCoV enters cells via the endocytic and exogenous protease-mediated pathways simultaneously. Integrins (ITG) αVß3 is a host factor required for PDCoV infection, especially during virus adhesion, invasion, and release. Most importantly, PDCoV promotes viral infection by activating the ITGαVß3-focal adhesion kinase (FAK)-phosphatidylinositol 3-kinase (PI3K)-serine/threonine-specific protein kinase (AKT) signalling pathway and induces inflammation by activating the ITGαVß3-FAK-PI3K-AKT-NF-κB signalling pathway. Overall, this is the first study to identify ITGαVß3 as an essential factor for PDCoV infection, which can help us to confirm the molecular regulatory mechanism and provide a comprehensive resource for PDCoV infection.

Unusual Migration of Bullet from the Right Scalp to Left Pyriform Fossa.

Spontaneous migration of a retained bullet is rare. Foreign bodies like bullets will be found at the distant site from the region of wound of entry in case they migrate. We report a case of a 17-year-old female with spontaneous migration of bullet from right temporo- parietal scalp to the posterior wall of left pyriform fossa. Twelve years back she had received a gunshot in right temporal region, no exploration was done to remove the bullet. She started complaining of discomfort in the upper neck of two months duration. Rigid laryngoscopy revealed smooth bulge with thinning of overlying mucosa. Radiographs of the neck showed bullet along the posterior pharyngeal wall in the region of the pyriform fossa. CT scan of the head and neck region was done to exclude the presence of any other foreign bodies. Microlaryngoscopy suspension of the larynx was done and CO2 laser was used to incise the mucosa over the bullet and it was retrieved with the help of long artery forceps. Our case illustrates that bullets may take an unexpected course from the site of entry, probably by migration. Trans oral CO2 laser microsurgery is the recommended approach for removal of such foreign bodies.

Anatomoradiological comparison between the minipterional and supraorbital eyebrow approaches to the interpeduncular region.

OBJECTIVE: Advances in surgical technology and microneurosurgery have led to increased utilization of so-called minimally invasive approaches, including the supraorbital eyebrow (SE) and minipterional (MPT) approaches for lesions involving the interpeduncular region. This study aimed to describe and compare anatomical landmarks, along with highlighting the advantages and disadvantages of the SE and MPT approaches to the interpeduncular region. METHODS: Ten formalin-fixed, latex-injected cadaveric specimens were used to perform bilateral SE and MPT approaches to the interpeduncular region. The operative depth of each approach to key anatomical landmarks was measured. Forty-five axial thin-slice computed tomography studies were reviewed to calculate the operative angles, with consideration of the midline as a reference. A 3D interactive anatomical model generated through the photogrammetry scanning technique was described. RESULTS: The depths of the operative corridors of the SE and MPT approaches to the interpeduncular fossa were 83.4 ± 1.8 mm and 67.7 ± 3.2 mm, respectively (p < 0.001). The mean angle of the MPT approach to the interpeduncular fossa was significantly wider than the one provided by the SE approach (39.9° ± 5.1° vs 28.4° ± 3.6°, p < 0.001). The interpeduncular region can consistently be accessed through the carotid-oculomotor triangle with the SE approach, as well as with the MPT approach. Furthermore, the SE route offered adequate access to the interpeduncular fossa through the opticocarotid triangle. The MPT route provided direct access to the upper prepontine cistern and anterior mesencephalic zone (AMZ). CONCLUSIONS: The MPT approach provides a wider and shorter operative corridor and can be employed for lesions in the interpeduncular region with extension to the prepontine cistern and ventrolateral midbrain lesions requiring access through the AMZ. The SE approach is better suited for ventromedial midbrain lesions requiring access via the interpeduncular fossa safe entry zone. Additional studies analyzing these approaches in a clinical setting will help to delineate their reliability and efficacy.

Polypyrimidine Tract-Binding Protein Enhances Zika Virus Translation by Binding to the 5' UTR of Internal Ribosomal Entry Site.

Objectives To identify the 5' untranslated region of Zika virus (ZIKV5'UTR) RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site (IRES) located in ZIKV5'UTR and virus production. Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining. Subsequently, liquid chromatography-tandem mass spectrometry (LC-MS/MS), bioinformatics analysis, and western blot were used to identify the candidate proteins binding to ZIKV5'UTR. Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production. Results tRSA RNA pull-down assay, LC-MS/MS, and western blot analysis showed that polypyrimidine tract-binding protein (PTB) bound to the ZIKV 5'UTR Furthermore, dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV (t = 10.220, P < 0.001), while PTB knockdown had the opposite effect (t = 4.897, P < 0.01). Additionally, virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer (t = 6.400, P < 0.01), whereas reducing PTB expression level weakened virus infectivity (t = 5.055, P < 0.01). Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.

New Therapies and Strategies to Curb HIV Infections with a Focus on Macrophages and Reservoirs.

More than 80 million people worldwide have been infected with the human immunodeficiency virus (HIV). There are now approximately 39 million individuals living with HIV/acquired immunodeficiency syndrome (AIDS). Although treatments against HIV infection are available, AIDS remains a serious disease. Combination antiretroviral therapy (cART), also known as highly active antiretroviral therapy (HAART), consists of treatment with a combination of several antiretroviral drugs that block multiple stages in the virus replication cycle. However, the increasing usage of cART is inevitably associated with the emergence of HIV drug resistance. In addition, the development of persistent cellular reservoirs of latent HIV is a critical obstacle to viral eradication since viral rebound takes place once anti-retroviral therapy (ART) is interrupted. Thus, several efforts are being applied to new generations of drugs, vaccines and new types of cART. In this review, we summarize the antiviral therapies used for the treatment of HIV/AIDS, both as individual agents and as combination therapies, and highlight the role of both macrophages and HIV cellular reservoirs and the most recent clinical studies related to this disease.

Improving United States HPV vaccination rates: Factors predictive of parental attitudes towards middle school entry requirements.

Although the human papillomavirus (HPV) vaccine is effective at preventing infection and certain types of cancer, uptake is suboptimal. HPV vaccine requirements for school entry are an underutilized strategy to increase HPV vaccine uptake among adolescents. The purpose of this study was to understand the factors that are predictive of parents' attitudes toward schools requiring the HPV vaccine for entry into middle school. Parents of adolescents ages 11-12 y were recruited to participate in an online survey via Qualtrics. Descriptive frequencies were obtained, and sequential regression analyses were conducted controlling for demographic characteristics. A total of 1,046 participants were included in the analysis. The mean age was 40.3 y (SD = 6.3) and the majority of participants were White (74.4%) and had some college education or higher (80.9%). Participant's gender, political affiliation, urban/rural setting, and education level were significantly associated with attitudes toward school entry requirements. Adding psychosocial items related to perceptions of benefits, risks, and social norms significantly increased the amount of variance explained in the model [(ΔR2 = .312, F(5, 1036) = 132.621)]. Perceived social norms was the strongest predictor of attitudes [ß = 0.321]. The results of this study can be used to inform policy changes around school-entry requirements in the United States. Further studies are needed to assess the influence of perceived social norms in vaccine hesitant groups.

MSC-extracellular vesicle microRNAs target host cell-entry receptors in COVID-19: in silico modeling for in vivo validation.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has created a global pandemic with significant morbidity and mortality. SARS-CoV-2 primarily infects the lungs and is associated with various organ complications. Therapeutic approaches to combat COVID-19, including convalescent plasma and vaccination, have been developed. However, the high mutation rate of SARS-CoV-2 and its ability to inhibit host T-cell activity pose challenges for effective treatment. Mesenchymal stem cells (MSCs) and their extracellular vesicles (MSCs-EVs) have shown promise in COVID-19 therapy because of their immunomodulatory and regenerative properties. MicroRNAs (miRNAs) play crucial regulatory roles in various biological processes and can be manipulated for therapeutic purposes. OBJECTIVE: We aimed to investigate the role of lyophilized MSC-EVs and their microRNAs in targeting the receptors involved in SARS-CoV-2 entry into host cells as a strategy to limit infection. In silico microRNA prediction, structural predictions of the microRNA-mRNA duplex, and molecular docking with the Argonaut protein were performed. METHODS: Male Syrian hamsters infected with SARS-CoV-2 were treated with human Wharton's jelly-derived Mesenchymal Stem cell-derived lyophilized exosomes (Bioluga Company)via intraperitoneal injection, and viral shedding was assessed. The potential therapeutic effects of MSCs-EVs were measured via histopathology of lung tissues and PCR for microRNAs. RESULTS: The results revealed strong binding potential between miRNA‒mRNA duplexes and the AGO protein via molecular docking. MSCs-EVs reduced inflammation markers and normalized blood indices via the suppression of viral entry by regulating ACE2 and TMPRSS2 expression. MSCs-EVs alleviated histopathological aberrations. They improved lung histology and reduced collagen fiber deposition in infected lungs. CONCLUSION: We demonstrated that MSCs-EVs are a potential therapeutic option for treating COVID-19 by preventing viral entry into host cells.

The Pentamer glycoprotein complex inhibits viral Immediate Early transcription during Human Cytomegalovirus infections.

The Pentamer complex of Human Cytomegalovirus (HCMV) consists of the viral glycoproteins gH, gL, UL128, UL130, and UL131 and is incorporated into infectious virions. HCMV strains propagated extensively in vitro in fibroblasts carry UL128, UL130, or UL131 alleles that do not make a functional complex and thus lack Pentamer function. Adding functional Pentamer to such strains decreases virus growth in fibroblasts. Here, we show that the Pentamer inhibits productive HCMV replication in fibroblasts by repressing viral Immediate Early (IE) transcription. We show that ectopic expression of the viral IE1 protein, a target of Pentamer-mediated transcriptional repression, complements the growth defect of a Pentamer-positive virus. Furthermore, we show that the Pentamer also represses viral IE transcription in cell types where HCMV in vitro latency is studied. Finally, we identify UL130 as a functional subunit of the Pentamer for IE transcriptional repression and demonstrate that cyclic AMP Response Element (CRE) and NFkB sites within the Major Immediate Early Promoter that drives IE1 transcription contribute to this repression. We conclude that the HCMV Pentamer represses viral IE transcription.

Mechanistic Insights Into an Ancient Adenovirus Precursor Protein VII Show Multiple Nuclear Import Receptor Pathways.

Adenoviral pVII proteins are multifunctional, highly basic, histone-like proteins that can bind to and transport the viral genome into the host cell nucleus. Despite the identification of several nuclear localization signals (NLSs) in the pVII protein of human adenovirus (HAdV)2, the mechanistic details of nuclear transport are largely unknown. Here we provide a full characterization of the nuclear import of precursor (Pre-) pVII protein from an ancient siadenovirus, frog siadenovirus 1 (FrAdV1), using a combination of structural, functional, and biochemical approaches. Two strong NLSs (termed NLSa and NLSd) interact with importin (IMP)ß1 and IMPα, respectively, and are the main drivers of nuclear import. A weaker NLS (termed NLSb) also contributes, together with an additional signal (NLSc) which we found to be important for nucleolar targeting and intranuclear binding. Expression of wild-type and NLS defective derivatives Pre-pVII in the presence of selective inhibitors of different nuclear import pathways revealed that, unlike its human counterpart, FrAdV1 Pre-pVII nuclear import is dependent on IMPα/ß1 and IMPß1, but not on transportin-1 (IMPß2). Clearly, AdVs evolved to maximize the nuclear import pathways for the pVII proteins, whose subcellular localization is the result of a complex process. Therefore, our results pave the way for an evolutionary comparison of the interaction of different AdVs with the host cell nuclear transport machinery.

Assessment of the re-entry tear index as a prognostic indicator for renal perfusion improvement after thoracic endovascular aortic repair in patients with acute and subacute Stanford type b aortic dissection with renal artery involvement.

Background: Aortic dissection is the most common acute aortic syndrome, and renal artery is the most common involved artery. The size and location of the re-entry tear directly affect the blood flow enhancement of the false lumen branch artery after surgery. In this study, the morphology and hemodynamics of the re-entry tear were comprehensively analyzed, and the location and size of the re-entry tear were quantitatively evaluated to calculate the re-entry tear index (RTI). This study aimed to assess the predictive capability of a comprehensive quantitative RTI for improvement in renal perfusion following thoracic endovascular aortic repair (TEVAR) in cases of acute and subacute Stanford type B aortic dissection with renal artery involvement. Methods: In this prospective cohort study, 137 patients diagnosed with acute or subacute type B aortic dissection with concomitant renal artery involvement who underwent TEVAR at Anzhen Hospital in Beijing from October 2017 to November 2021 were enrolled. Renal blood flow was estimated quantitatively with ultrasound. Based on the ultrasound findings of renal artery flow, the patients were classified into two groups: group A [postoperative volume flow (VolFlow) reduced compared to preoperative VolFlow] and group B (postoperative VolFlow increased compared to preoperative VolFlow). All re-entry tears present in the aortic trunk according to reconstructed computed tomography angiography (CTA) obtained preoperatively were included in the analysis. The general information of patients, whether the involved renal artery arose partially or wholly from the false lumen, the proximal diameter and length of the covered stent, the diameter of primary entry tear, the RTI, etc. were analyzed. Univariate and multivariate logistic regression analyses were executed to assess the risk factors associated with increased renal arterial blood flow subsequent to TEVAR. Additionally, receiver operating characteristic (ROC) curve analysis was used to ascertain the optimal cutoff value and predictive efficacy of the RTI. Results: A total of 137 patients, comprising of 32 with acute and 105 with subacute type B aortic dissection accompanied by renal artery involvement, underwent TEVAR. Among these patients, 44 (32.1%) were assigned to group A and 93 (67.9%) to group B. Renal blood flow exhibited an increase in 67.9% of the patients after TEVAR. The results of multivariate analysis indicated that the RTI is an independent risk factor for postoperative renal perfusion improvement [odds ratio =17.66; 95% confidence interval (CI): 2.13-78.55; P=0.020]. The optimal cutoff value for RTI, determined to be 0.033, demonstrated the ability to identify renal perfusion improvement in patients without hypertension with a sensitivity of 53.7% and a specificity of 68.9%. In patients with concomitant hypertension, RTI exhibited a sensitivity of 96.6% and a specificity of 60.0%, with an area under the ROC curve (AUC) of 0.792 (95% CI: 0.643-0.941; P=0.021) for identifying renal perfusion improvement. Conclusions: RTI demonstrated a favorable predictive value for improving renal malperfusion following TEVAR in cases of aortic dissection with renal artery involvement.

Risk factors for proximal and distal aortic events after type A acute aortic dissection.

OBJECTIVES: Type A acute aortic dissection (TAAAD) is a life-threatening condition often requiring emergency surgery, with approximately 30% of patients needing reoperation. This study aimed to identify predictors of long-term aortic events from early postoperative computed tomography (CT) examinations. METHODS: A total of 336 cases underwent TAAAD surgery at two institutions between 2002 and 2018. Of these, 302 patients received CT examinations immediately after initial TAAAD surgery. Predictors of aortic events were evaluated from these early postoperative CT exams. Aortic events were defined as any events involving aortic-related death, open surgery, reoperation, endovascular stenting, or thoracic aorta diameter enlargement to ≥ 55 mm. RESULTS: Excluding 34 in-hospital deaths (10.1%; 34/336), the 1-, 5-, and 10-year actuarial survival rates after primary TAAAD surgery were 98.2%, 88.6%, and 81.7%, respectively. Over a mean follow-up period of 7.4 ± 5.1 years, 67 aortic events (proximal: 19, distal: 45, both: 3) were observed. Freedom from proximal aortic events was 98.6%, 93.9%, and 85.2% at 1, 5, and 10 years, respectively. Proximal anastomosis new entry was identified as a significant risk factor for aortic events, with a 92% vs. 42% incidence at 10 years (p < 0.001). Freedom from distal aortic events was 99.6%, 84.5%, and 67.2% at 1, 5, and 10 years, respectively. A false/true area ratio greater than 1 and distal anastomosis new entry were significant risk factors for aortic events (low-risk group: 83.3% vs. high-risk group: 42.3% at 10 years, p < 0.001). CONCLUSIONS: Detailed analysis of early postoperative CT scans following primary TAAAD surgery may help identify predictors of subsequent aortic events, potentially improving long-term patient management and outcomes.

Differential functional role of Orai1 variants in constitutive Ca2+ entry and calcification in luminal breast cancer cells.

Resting cytosolic Ca2+ concentration is tightly regulated to fine-tune Ca2+-dependent cellular functions. Luminal breast cancer cells exhibit constitutive Ca2+ entry mediated by Orai1 and the secretory pathway Ca2+-ATPase, SPCA2, which result in mammary microcalcifications that constitute a prognostic marker of mammary lesions. Two Orai1 isoforms have been identified, the full-length Orai1α, consisting of 301 amino acids, and the short variant, Orai1ß, lacking the 63 or 70 N-terminal amino acids comprising residues involved in channel inactivation and binding sites with Orai1 partners. We show that only the mammalian-specific Orai1α rescues SPCA2-dependent constitutive Ca2+ entry in Orai1-KO MCF7 cells, a widely used luminal breast cancer cell line. FRET analysis and immunoprecipitation revealed that Orai1α shows a greater ability to interact with SPCA2 than Orai1ß. Deletion of the first 38 amino acids in Orai1α reduced the interaction with SPCA2 to a similar extent as Orai1ß, thus suggesting that the N-terminal 38 amino acids play a relevant role in Orai1α-SPCA2 interaction. Finally, Orai1α, but not Orai1ß, rescue the ability of Orai1-deficient cells to form in vitro microcalcifications. These findings provide compelling evidence for a functional role of Orai1α in constitutive Ca2+ entry in MCF7 cells, which might be a target to prevent the development of mammary microcalcifications in luminal breast cancer.

Evaluating Perceptions of Head and Neck Surgeons on the Role of Single-Entry Models in Managing Surgical Waitlists in Ontario: A Qualitative Study.

BACKGROUND: Long surgical wait times have long plagued health systems in Canada and abroad. This backlog and associated strain on health human resources has been exacerbated by the COVID-19 pandemic, affecting surgeries of varying degrees of urgency across all surgical specialties, including head and neck surgery. Single-entry models (SEMs) are being increasingly studied as one possible strategy to help manage surgical wait times, and a growing number of health systems have implemented SEMs within departments such as otolaryngology-head and neck surgery. We sought to evaluate the views of head and neck surgeons at all 8 designated head and neck cancer centers across Ontario on the role of SEMs in managing surgical backlogs. RESULTS: We interviewed 10 Ontario head and neck surgeons on the role of SEMs in managing wait times within the field. The following themes were elicited from interview transcripts: (1) anticipated positive impact, (2) barriers to implementation, (3) patient experience, and (4) roadmap to implementation. Participants agreed that SEMs may have utility for certain types of surgeries if implemented to address local needs. They also believe this model would have the greatest impact if employed together with other approaches, such as increasing operating room time or nursing availability. CONCLUSION: Our results highlighted the necessity for a nuanced approach to single-entry model implementation in head and neck surgery. While participants recognized the utility of SEMs for high-volume and low-variation surgeries, participants remained divided on the optimal approach to triaging patients necessitating more complex oncologic treatments. Deliberate collaboration among stakeholder organizations and senior surgeons will be critical if SEMs are to succeed in an intricate and political healthcare environment.

Effectiveness and safety of Tachosil® as a ventricular sealant: an observational cohort study.

PURPOSE: Surgery close to or in contact with the ventricular system is challenging due to the complications. We sought to evaluate the effectiveness and safety of TachoSil® as a ventricular sealant in preventing complications after cranial surgery with an open ventricular system (OVS). METHODS: This is a single-center and prospective cohort study We included patients who underwent elective surgery for supratentorial craniotomy and periventricular pathology between December 2020 and November 2023. We registered surgical complications arising from CSF dynamics (such as percutaneous cerebrospinal fluid (CSF) leakage, hydrocephalus, pseudomeningocele), infections, and other complications (postsurgical hematoma) adverse drug reactions (ADRs), reintervention or hospital readmission up to 90 days after surgery. RESULTS: Forty interventions were performed on 39 patients, whose median age was 56 years. Eleven patients (28.2%) had antecedents of previous surgery in the same location, 5 (12.8%) had previously received radiotherapy and chemotherapy, and 11 (28.2%) were smokers. Twenty-four patients (60%) underwent surgery for high-grade glioma, 8 (20%) for low-grade gliomas, 6 (15%) for metastasis and 2 (5%) for meningioma. Throughout the study and up to 90 days after surgery, none of the patients presented an ADR. Only 2 patients (5%) presented with a surgery complications derived from ventricular opening (one patient with a percutaneous CSF leakage and one patients with external hydrocephalus). Both patients resolved with a ventriculoperitoneal shunt. CONCLUSIONS: TachoSil® is a dural sealant that can be used safely and effectively intraparenchymally in patients whose surgery involves a ventricular opening. Only 5% of treated patients presented complications arising from CSF hydrodynamics. No patients had pseudomeningocele, infections or complications related to the use of this sealant. To confirm these positive results, randomized and comparative clinical trials assessing the efficacy of TachoSil® in patients after cranial surgery with an OVS are essential. TRIAL REGISTRATION NUMBER AND DATE OF REGISTRATION: This study was registered in the Clinical Trials.gov (NCT05717335). Date May 1st, 2022.

Virtual Waiting Room: The New Narrative of Waiting in Oncology Care.

This conceptual study introduces the "virtual waiting room," an innovative, interactive, web-based platform designed to enhance the waiting experience in oncology by providing personalized, educational, and supportive content. Central to our study is the implementation of the circular entry model, which allows for non-linear navigation of health information, empowering patients to access content based on their immediate needs and interests. This approach respects the individual journeys of patients, acknowledging the diverse pathways through which they seek understanding and manage their health. The virtual waiting room is designed not only to support patients but also to facilitate stronger communication and shared understanding between patients, caregivers, and families. By providing a shared digital space, the platform enables caregivers and family members to access the same information and resources, thereby promoting transparency and collective knowledge. This shared access is crucial in managing the emotional complexities of oncology care, where effective communication can significantly impact treatment outcomes and patient well-being. Furthermore, the study explores how the circular entry model within the virtual waiting room can enhance patient autonomy and engagement by offering customized interactions based on user feedback and preferences. This personalized approach aims to reduce anxiety, improve health literacy, and prepare patients more effectively for clinical interactions. By transforming passive waiting into active engagement, the virtual waiting room turns waiting time into a meaningful, informative period that supports both the psychological and informational needs of patients and their support networks.

Differential regulation of viral entry-associated genes modulated by inflammatory cytokines in the nasal epithelium.

This study aimed to investigate the impact of different types of nasal inflammation on the regulation of entry-associated genes of respiratory viruses, including severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), Middle East respiratory syndrome coronavirus (MERS-CoV), human coronavirus 229E (HCoV-229E), and influenza virus, in the nasal epithelium. Subjects were classified into three groups: control, eosinophilic chronic rhinosinusitis (ECRS), and noneosinophilic CRS (NECRS) groups. Angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine subtype 2 (TMPRSS2), alanyl aminopeptidase (ANPEP), dipeptidyl peptidase 4 (DPP4), and beta-galactoside alpha-2,6-sialyltransferase 1 (ST6GAL1), and beta-galactoside alpha-2,3-sialyltransferase 4 (ST3GAL4) were selected as key entry-associated genes for SARS-CoV-2, HCoV-229E, MERS-CoV, and influenza, respectively, and were evaluated. Brushing samples obtained from each group and human nasal epithelial cells cultured using an air-liquid interface system were treated for 7 days with typical inflammatory cytokines and analyzed using real-time polymerase chain reaction. Western blot analysis and confocal microscopy were performed. The entry-associated genes showed distinct regulation patterns in response to each interleukin-4 (IL-4), interleukin-13 (IL-13), tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ). Specifically, ACE2 significantly decreased in type 2 cytokines (IL-4 and IL-13), while TMPRSS2 significantly decreased in type 1 cytokines (TNF-α and IFN-γ). ANPEP significantly decreased in both types of cytokines. Remarkably, DPP4 significantly increased in type 2 cytokines and decreased in type 1 cytokines. Moreover, ST6GAL1 and ST3GAL4 significantly increased in type 2 cytokines and decreased in type 1 cytokines, particularly IFN-γ. These findings were supported by western blot analysis and confocal imaging results, especially for ACE2 and DPP4. The findings regarding differential regulation suggest that patients with ECRS, primarily mediated by type 2 inflammation, may have lower susceptibility to SARS-CoV-2 and HCoV-229E infections but higher susceptibility to MERS-CoV and influenza infections.

STIM Proteins: The Gas and Brake of Calcium Entry in Neurons.

Stromal interaction molecules (STIM)s are Ca2+ sensors in internal Ca2+ stores of the endoplasmic reticulum. They activate the store-operated Ca2+ channels, which are the main source of Ca2+ entry in non-excitable cells. Moreover, STIM proteins interact with other Ca2+ channel subunits and active transporters, making STIMs an important intermediate molecule in orchestrating a wide variety of Ca2+ influxes into excitable cells. Nevertheless, little is known about the role of STIM proteins in brain functioning. Being involved in many signaling pathways, STIMs replenish internal Ca2+ stores in neurons and mediate synaptic transmission and neuronal excitability. Ca2+ dyshomeostasis is a signature of many pathological conditions of the brain, including neurodegenerative diseases, injuries, stroke, and epilepsy. STIMs play a role in these disturbances not only by supporting abnormal store-operated Ca2+ entry but also by regulating Ca2+ influx through other channels. Here, we review the present knowledge of STIMs in neurons and their involvement in brain pathology.

Envelope Protein-Targeting Zika Virus Entry Inhibitors.

Zika virus (ZIKV; family, Flaviviridae), which causes congenital Zika syndrome, Guillain-Barré Syndrome, and other severe diseases, is transmitted mainly by mosquitoes; however, the virus can be transmitted through other routes. Among the three structural and seven nonstructural proteins, the surface envelope (E) protein of ZIKV plays a critical role in viral entry and pathogenesis, making it a key target for the development of effective entry inhibitors. This review article describes the life cycle, genome, and encoded proteins of ZIKV, illustrates the structure and function of the ZIKV E protein, summarizes E protein-targeting entry inhibitors (with a focus on those based on natural products and small molecules), and highlights challenges that may potentially hinder the development of effective inhibitors of ZIKV infection. Overall, the article will provide useful guidance for further development of safe and potent ZIKV entry inhibitors targeting the viral E protein.

A comparative study of the versatility of various entry points for double-puncture TMJ arthrocentesis: A randomized controlled study.

Although various authors employed various entry points for the double-puncture technique (DPT) for arthrocentesis, the literature is devoid of any comparative studies. Therefore, the current prospective study aimed to evaluate the versatility of these different points. A total of 144 TMJs in 108 patients were included and randomly divided into two categories according to ID stage: category I (disc displacement without reduction with limitation), and category II (disc displacement without reduction without limitation). Patients in every category were randomly divided into 3 groups according to the site of entry point of the second needle: group 1 (20-10 point: 20 mm anterior to the tragus and 2 mm inferior to the cantho-tragus line), group 2 (20-1 point), and group 3 (7-2 point). For all patients, the first entry point was 10-2, and the upper joint cavity was irrigated with 150 ml of Ringer's solution without subsequent intra-articular injections. Group 3 had better results than group 2 and further than group 1 in categories I and II with regard to the number of second needle relocations, ease of the procedure, duration of the procedure, and nature of the outflow, as well as pain at rest and pain on function at 1, 3, and 6 post-operative months. For the maximum mouth opining, group 3 had better results than group 2 and further than group 1 only in category I. Therefore, techniques depending on the superior posterior entry points (such as 7-2 point) were recommended.

SARS-CoV-2 Omicron subvariants progressively adapt to human cells with altered host cell entry.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant exhibits high transmissibility with a strong immune escape ability and causes frequent large-scale global infections by producing predominant subvariants. Here, using human upper/lower airway and intestinal cells, we examined the previously dominant BA.1-BA.5 and BA.2.75 subvariants, together with the recently emerged XBB/BQ lineages, in comparison to the former Delta variant. We observed a tendency for each virus to demonstrate higher growth capability than the previously dominant subvariants. Unlike human bronchial and intestinal cells, nasal epithelial cells accommodated the efficient entry of certain Omicron subvariants, similar to the Delta variant. In contrast to the Delta's reliance on cell-surface transmembrane protease serine 2, all tested Omicron variants depended on endosomal cathepsin L. Moreover, S1/S2 cleavage of early Omicron spikes was less efficient, whereas recent viruses exhibit improved cleavage efficacy. Our results show that the Omicron variant progressively adapts to human cells through continuous endosome-mediated host cell entry.IMPORTANCESARS-CoV-2, the causative agent of coronavirus disease 2019, has evolved into a number of variants/subvariants, which have generated multiple global waves of infection. In order to monitor/predict virological features of emerging variants and determine appropriate strategies for anti-viral development, understanding conserved or altered features of evolving SARS-CoV-2 is important. In this study, we addressed previously or recently predominant Omicron subvariants and demonstrated the gradual adaptation to human cells. The host cell entry route, which was altered from the former Delta variant, was conserved among all tested Omicron subvariants. Collectively, this study revealed both changing and maintained features of SARS-CoV-2 during the Omicron variant evolution.