Virion-associated influenza hemagglutinin clusters upon sialic acid binding visualized by cryo-electron tomography.

Influenza viruses are enveloped, negative sense single-stranded RNA viruses covered in a dense layer of glycoproteins. Hemagglutinin (HA) accounts for 80-90% of influenza glycoprotein and plays a role in host cell binding and membrane fusion. While previous studies have characterized structures of receptor-free and receptor-bound HA in vitro, the effect of receptor binding on HA organization and structure on virions remains unknown. Here, we used cryo-electron tomography (cryoET) to visualize influenza virions bound to a sialic acid receptor mimic. Overall, receptor binding did not result in significant changes in viral morphology; however, we observed rearrangements of HA trimer organization and orientation. Compared to the even inter-glycoprotein spacing of unliganded HA trimers, receptor binding promotes HA trimer clustering and formation of a triplet of trimers. Subtomogram averaging and refinement yielded 8-10 Å reconstructions that allowed us to visualize specific contacts between HAs from neighboring trimers and identify molecular features that mediate clustering. Taken together, we present new structural evidence that receptor binding triggers clustering of HA trimers, revealing an additional layer of HA dynamics and plasticity.

Amputation Triggers Long-Range Epidermal Permeability Changes in Evolutionarily Distant Regenerative Organisms.

Previous studies have reported that amputation invokes body-wide responses in regenerative organisms, but most have not examined the implications of these changes beyond the region of tissue regrowth. Specifically, long-range epidermal responses to amputation are largely uncharacterized, with research on amputation-induced epidermal responses in regenerative organisms traditionally being restricted to the wound site. Here, we investigate the effect of amputation on long-range epidermal permeability in two evolutionarily distant, regenerative organisms: axolotls and planarians. We find that amputation triggers a long-range increase in epidermal permeability in axolotls, accompanied by a long-range epidermal downregulation in MAPK signaling. Additionally, we provide functional evidence that pharmacologically inhibiting MAPK signaling in regenerating planarians increases long-range epidermal permeability. These findings advance our knowledge of body-wide changes due to amputation in regenerative organisms and warrant further study on whether epidermal permeability dysregulation in the context of amputation may lead to pathology in both regenerative and non-regenerative organisms.

Network meta-analysis on the mechanisms underlying alcohol augmentation of acute pancreatitis and diabetes type II.

BACKGROUND: Pancreatitis is a severe inflammatory pathology that occurs from pancreatic duct and exocrine acinar injury, leading to improper secretion of digestive enzymes, auto-digestion of the pancreas, and subsequent inflammation. Clinical reports show that 60%-90% of pancreatitis patients have a history of chronic alcohol use. More recent studies reveal that exocrine pancreas disorders like acute pancreatitis can precede diabetes type II onset, though mechanisms are not yet fully known. This study identified molecules and key signaling pathways underlying alcohol-induced acute pancreatitis and their effects on diabetes type II onset. METHODS: Data on human peripheral blood samples with or without acute pancreatitis were retrieved from the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (accession number GSE194331). Acute pancreatitis-mediated differentially expressed genes (DEGs) were generated from GSE194331 using CLC Genomics Workbench 12. Molecules associated with ethanol (EtOH), acute pancreatitis, and diabetes type II were collected from QIAGEN Knowledge Base (QKB). The relationship between the molecules and signaling pathways associated with EtOH, acute pancreatitis, or diabetes type II was examined using various Ingenuity Pathway Analysis (IPA) tools. RESULTS: Our investigation showed that acute pancreatitis-mediated DEGs were closely associated with EtOH by revealing that EtOH-induced acute pancreatitis appears to lead to the onset of diabetes type II. We found that diabetes type II onset was mediated by pro-inflammatory and metabolic mechanisms underlying EtOH-induced acute pancreatitis, involving increased expression of cytokines including macrophage migration inhibitory factor (MIF), and decreased expression of hormones such as insulin. CONCLUSIONS: Exposure to alcohol may promote diabetes type II by affecting the activity of key inflammatory and metabolic mediators involved in acute pancreatitis. These findings call for further investigation into the role of pro-inflammatory and metabolic mediators like resistin, IL-6, and insulin in EtOH-induced diabetes type II associated with acute pancreatitis pathologies.

Putative epithelial-mesenchymal transitions during salamander limb regeneration: Current perspectives and future investigations.

Previous studies have implicated epithelial-mesenchymal transition (EMT) in salamander limb regeneration. In this review, we describe putative roles for EMT during each stage of limb regeneration in axolotls and other salamanders. We hypothesize that EMT and EMT-like gene expression programs may regulate three main cellular processes during limb regeneration: (1) keratinocyte migration during wound closure; (2) transient invasion of the stump by epithelial cells undergoing EMT; and (3) use of EMT-like programs by non-epithelial blastemal progenitor cells to escape the confines of their niches. Finally, we propose nontraditional roles for EMT during limb regeneration that warrant further investigation, including alternative EMT regulators, stem cell activation, and fibrosis induced by aberrant EMT.

Antitumor activity of afatinib in EGFR T790M-negative human oral cancer therapeutically targets mTOR/Mcl-1 signaling axis.

PURPOSE: This study investigates the role and effectiveness of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) in oral cancer, focusing on the clinical relevance of EGFR and myeloid cell leukemia-1 (Mcl-1) in head and neck cancers (HNCs). It aims to explore the molecular mechanism of afatinib, a TKI, in treating human oral cancer. METHODS: We conducted an in silico analysis using databases like The Cancer Genome Atlas, Gene Expression Omnibus, and Clinical Proteomic Tumor Analysis Consortium, along with immunohistochemistry staining, to study EGFR and Mcl-1 expression in HNCs. For investigating afatinib's anticancer properties, we performed various in vitro and in vivo analyses, including trypan blue exclusion assay, Western blotting, 4'-6-diamidino-2-phenylindole staining, flow cytometry, quantitative real-time PCR, Mitochondrial membrane potential assay, overexpression vector construction, transient transfection, and a tumor xenograft model. RESULTS: Higher expression levels of EGFR and Mcl-1 were observed in HNC patient tissues compared to normal tissues, with their co-expression significantly linked to poor prognosis. There was a strong correlation between EGFR and Mcl-1 expressions in oral cancer patients. Afatinib treatment induced apoptosis and suppressed Mcl-1 in oral cancer cell lines without the EGFR T790M mutation. The mechanism of afatinib-induced apoptosis involved the EGFR/mTOR/Mcl-1 axis, as shown by the effects of mTOR activator MHY1485 and inhibitor rapamycin. Afatinib also increased Bim expression, mitochondrial membrane permeabilization, and cytochrome c release. It significantly lowered tumor volume without affecting body, liver, and kidney weights. CONCLUSION: Afatinib, targeting the EGFR/mTOR/Mcl-1 axis, shows promise as a therapeutic strategy for oral cancer, especially in patients with high EGFR and Mcl-1 expressions.

Management of Recurrent Giant Cell Tumor of Distal Tibia With Intramedullary Hindfoot Fusion Nail and Trabecular Metal Implant Construct.

Reconstruction options for giant cell tumors (GCTs) of bone are limited and challenging due to the amount of structural compromise and the high recurrence rates. This is especially true for GCTs of the foot and ankle, as the area is vital for weight bearing and function. The typical treatment for GCTs is currently excision, curettage, and cementation, although that is not always effective. A 36-year-old otherwise healthy female presented with an original diagnosis of a large aneurysmal bone cyst (ABC) of the distal tibia that had recurred despite two previous attempts at treatment with resection and cementation. She was treated with surgical resection of the lesion, reconstruction, and ankle and subtalar joint arthrodesis with a tibiotalocalcaneal intramedullary nail in combination with a trabecular metal cone. The final pathology of the intraoperative samples was consistent with GCT. Postoperatively, she recovered well, and her imaging was consistent with a successful fusion. This case report provides evidence that tibiotalocalcaneal fusion with a unique combination of hindfoot nail and trabecular metal cone construct in a single procedure is a successful option for the treatment of large, recurrent GCT lesions in the distal tibia.

Food impaction on three-dimensional printed models with periodontal ligament simulation.

OBJECTIVE: This study aims to evaluate food impaction on three-dimensional (3D) printed models with periodontal ligament simulation. MATERIALS AND METHODS: Based on a commercial typodont pair, 3D maxillary and mandibular models were created with no teeth and with tooth sockets that were 1 mm wider than the original ones from 24 to 27 or 34 to 37 for periodontal ligament simulation with vinyl polysiloxane impression material. In total, 35 pairs of 7 combinations, including maxillary/mandibular typodonts in occlusion with maxillary/mandibular 3D models with/without a distal gap of canines on 3D models (tooth 23 or 33) were mounted on hinge articulators and divided into seven groups (n = 5). Each sample experienced the same manual chewing simulation on a customized device. The proximal surfaces were photographed to measure the percentage of food impaction area using ImageJ software. RESULTS: Group with fixed maxillary and mandibular teeth showed more food impaction than other groups with significant differences in the average of maxilla and the average of all proximal areas. CONCLUSION: The flexibility of the periodontal ligament and the degree of freedom of the teeth in their sockets may contribute to the extent of food impaction in proximal spaces.

Effects of dentin bonding agents and silanization on bond strength between 3D printed resin and composite resin.

This study aimed to evaluate the effects of dentin bonding agents and silanization on the bond strength between 3D printed resin and composite resin and compare it with a conventional composite resin. 3D printed resin cylinders (PCB) and composite resin substrates (Z250) were prepared and divided into eight subgroups based on the bonding agents used (n=12). The shear bond strength was measured using a universal testing machine, and the failure modes were evaluated. The bond strength was found to vary significantly among the bonding agents and substrate types. Silane application did not significantly improve the bond strength. Among the bonding agents, the universal adhesives exhibited the highest bond strengths for both substrates. Compared to PCB, Z250 demonstrated stronger bonds and exhibited more cohesive failures. Further research is needed to optimize the surface treatments and resin formulations for enhanced bond strength and durability between 3D printed and composite resins.

Trueness, physical properties, and surface characteristics of additive-manufactured zirconia crown.

OBJECTIVE: This study aimed to conduct a comparison of trueness and physical and surface properties among five distinct types of additive manufactured (AM) zirconia crowns and zirconia crowns produced using the subtractive manufacturing (SM). MATERIAL AND METHODS: Zirconia crowns were fabricated using five distinct techniques, each varying in the method of slurry transfer and photocuring source. Each experimental group utilized either one of the four digital light processing (DLP)-based techniques (DLP spreading, DLP spreading gradation, DLP vat and DLP circular spreading) or the stereolithography (SLA)-based technique (SLA spreading). The control (CON) group employed SM. To assess accuracy, trueness was measured between the scan and reference data. To analyze the physical properties, voids were examined using high-energy spiral micro-computed tomography scans, and the crystal structure analysis was performed using X-ray diffraction (XRD). Surface roughness was assessed through laser scanning microscopy. RESULTS: Differences in the trueness of internal surfaces of crowns were found among the groups (P < 0.05). Trueness varied across the measurement surfaces (occlusal, lateral, and marginal) in all the groups except for the DLP spreading gradation group (P < 0.05). Voids were observed in all AM groups. All groups showed similar XRD patterns. All AM groups showed significantly greater surface roughness compared to the CON group (P < 0.001). CONCLUSION: The AM zirconia crowns showed bubbles and a rougher surface compared to the SM crowns. All groups exhibited typical zirconia traits and trueness levels within clinically acceptable limits, suggesting that current zirconia AM techniques could be suitable for dental applications.

Post-polymerization of three-dimensional printing resin using a dental light curing unit.

Background/Purpose: In vat photopolymerization, post-polymerization of the three-dimensional (3D) printing resin is necessary to ensure the optimum physical properties of the printed objects. This study aimed to evaluate the potential use of a handheld polywave light-emitting diode (LED) dental light-curing unit (LCU) for post-polymerizing 3D printed resins by measuring the microhardness and biaxial flexural strength of the post-polymerized resin. Material and methods: 3D printed 1- and 2-mm-thick disks were irradiated with a dental LCU at 3200 mW/cm2. Post-polymerization was repeated either on one side from the top surface: two cycles (T2), four cycles (T4), and eight cycles (T8), or on both sides from the top and bottom surfaces: one cycle (T1B1), two cycles (T2B2), and four cycles (T4B4) for each side. The microhardness and biaxial strength of the disks were compared to those post-polymerized by a conventional desktop polymerizing unit (PC) and those without post-polymerization (NC). Results: Microhardness of the disks varied between the top and bottom surfaces of the 1-mm and 2-mm-thick disks, depending on the post-polymerization methods. T8 and T4B4 produced comparable microhardness on the top surface to PC for both thicknesses. In contrast, PC, T2B2, and T4B4 exhibited the highest microhardness on the bottom surface. Except for NC, the 1-mm-thick disks had a higher biaxial flexural strength than the 2-mm-thick disks. T4B4 resulted in the highest biaxial flexural strength for both thicknesses, which was comparable to that of the desktop polymerizing unit. Conclusion: The microhardness and biaxial flexural strengths of the post-polymerized 3D-printed disks increase with polymerization time. With sufficient polymerization from both sides, the polywave LCU has the potential to be a viable alternative to desktop polymerization units.

Determinants of Dentist-Patient Relationships: A Scoping Review.

OBJECTIVES: The dentist-patient relationship (DPR) is considered to be a key element in dental clinical settings. This scoping review aimed to examine the extent of previous research on DPR, focussing on its determinants for the reification of the construct. METHODS: This research was directed by the guidance for systematic scoping reviews from the Joanna Briggs Institute. The inclusion/exclusion criteria were based on participants of general adults and dentists, the concept of determinants of DPR, and the context of dental health care encounters. A literature search was performed in 6 major electronic databases in July 2023. Key information from included articles was extracted to chart the results, mainly to identify the determinants of DPR. Each determinant of DPR was classified according to the conceptual model of DPR. RESULTS: A total of 1727 records were initially identified, and 16 articles were included in the review. Nine studies used a quantitative method and 7 were nonempirical articles. All but 2 articles were from the perspective of patients. Factors were grouped into 6 main domains: dentist, patient, society/environment, clinical structure, clinical process, and outcome. Amongst the 6 domains of DPR, most determining factors were related to the clinical process. "Communication" was most frequently counted, at 8 times, followed by "trust" (frequency, 6). At the patient level, "dental fear/anxiety" was frequently used to measure DPR. CONCLUSIONS: Previous literature about DPR indicated a few common and dentistry-specific determinants from the patient perspective. Further studies are encouraged to develop a more comprehensive framework and evaluation scale of DPR.

Impaired sleep is associated with tau deposition on 18F-flortaucipir PET and accelerated cognitive decline, accounting for medications that affect sleep.

BACKGROUND: Impaired sleep is commonly associated with Alzheimer's disease (AD), although the underlying mechanisms remain unclear. Furthermore, the moderating effects of sleep-affecting medications, which have been linked to AD pathology, are incompletely characterized. Using data from the Alzheimer's Disease Neuroimaging Initiative, we investigated whether a medical history of impaired sleep, informant-reported nighttime behaviors, and sleep-affecting medications are associated with beta-amyloid and tau deposition on PET and cognitive change, cross-sectionally and longitudinally. METHODS: We included 964 subjects with 18F-florbetapir PET scans. Measures of sleep impairment and medication use were obtained from medical histories and the Neuropsychiatric Inventory Questionnaire. Multivariate models, adjusted for covariates, were used to assess associations among sleep-related features, beta-amyloid and tau, and cognition. Cortical tau deposition, categorized by Braak stage, was assessed using the standardized uptake value peak alignment (SUVP) method on 18F-flortaucipir PET. RESULTS: Medical history of sleep impairment was associated with greater baseline tau in the meta-temporal, Braak 1, and Braak 4 regions (p = 0.04, p < 0.001, p = 0.025, respectively). Abnormal nighttime behaviors were also associated with greater baseline tau in the meta-temporal region (p = 0.024), and greater cognitive impairment, cross-sectionally (p = 0.007) and longitudinally (p < 0.001). Impaired sleep was not associated with baseline beta-amyloid (p > 0.05). Short-term use of selective serotonin reuptake inhibitors and benzodiazepines slightly weakened the sleep-tau relationship. CONCLUSIONS: Sleep impairment was associated with tauopathy and cognitive decline, which could be linked to increased tau secretion from neuronal hyperactivity. Clinically, our results help identify high-risk individuals who could benefit from sleep-related interventions aimed to delay cognitive decline and AD.

Shear bond strength of dual-cured resin cements on zirconia: The light-blocking effect of a zirconia crown.

Abstract Background/purpose: The presence of restorative material between resin cement and the light-curing unit can reduce light transmission. This study aimed to evaluate the effect of the light-blocking effect of zirconia crown on shear bond strength (SBS) between three dual-cured resin cements and the zirconia surface. Materials and methods: Sixty zirconia specimens were prepared and divided into three groups according to the type of resin cement [RXU (RelyX Ultimate); SC2 (SmartCem2); MEC (Maxcem Elite Chroma)]. Each group was further divided into two subgroups, with or without a 1-mm-thick zirconia crown (n = 10). The specimens were light-cured from five different directions for 20 s each. All specimens were thermocycled 5000 times and subjected to SBS testing, followed by scanning electron microscope examination. Results: The presence of a 1-mm-thick zirconia crown had no significant effect on the SBS in all resin cements. However, the SBS was significantly affected by type of resin cement. RXU showed the highest SBS (8.35 MPa with crown; 8.57 MPa without crown), followed by SC2 (5.48 MPa with crown; 5.57 without crown) and then MEC (3.37 MPa with crown; 4.04 MPa without crown. Fractured surfaces exhibited varying degrees of mixed failure patterns. Conclusion: A 1-mm-thick zirconia crown material between the light source and the dual-cured resin cement did not significantly influence the SBS of the resin cements on the zirconia substrates. RXU exhibited the highest SBS regardless of zirconia crown coverage. With sufficient light-curing, dual-cured resin cements can be a good choice for zirconia crown cementation.

Investigating the role of chlorogenic acids and coffee type in coffee-induced teeth discoloration.

OBJECTIVE: Coffee is one of the most popular beverages in the world, with millions of people consuming it every day. The effect of coffee on teeth discoloration has long been a concern for both coffee drinkers and dental professionals. To address this concern, this study aimed to investigate the role of chlorogenic acids (CGAs) and the type of coffee in coffee-induced teeth discoloration. MATERIALS AND METHODS: High-performance liquid chromatography with a photodiode array detector was used to determine the CGA contents of instant coffee produced by five manufacturers (Starbucks, Dunkin' Donuts, Kanu, Ediya, Coffee Bean). A total of 180 bovine tooth specimens were immersed in the coffee samples for varying durations (3, 9, 24, 48, and 72 h), and the discoloration levels were measured using a spectrophotometer. A linear mixed-effects model analysis was used to determine the significance of L*, a*, and b* values in relation to the duration of coffee immersion and coffee type. RESULTS: Both immersion time and coffee type had significant effects on tooth discoloration (p < 0.001), with some types of coffee being more strongly associated with tooth discoloration than others. The amount of CGAs present in coffee was found to be positively correlated with the degree of discoloration (p = 0.030). CONCLUSIONS: Prolonged exposure to coffee can exacerbate teeth staining, and different types of coffee can cause varying degrees of discoloration. Furthermore, coffee with higher levels of CGAs may lead to greater tooth discoloration.

Assessing the efficacy of jet dispenser versus direct syringe injection for calcium hydroxide paste placement in artificial root canals.

This study aimed to evaluate the efficacy of a novel jet dispenser (JD) for filling simulated straight and curved canals with calcium hydroxide (Ca(OH)2) paste by comparing the level of intracanal medicament with traditional direct syringe delivery. The Ca(OH)2 paste was delivered into the canals using either a direct syringe with binding tip and unbinding tip or a JD. The analysis of the ratio of the canal filled area (RCFA) in the coronal, middle, and apical thirds was conducted using the Kruskal-Wallis and Mann-Whitney U tests (α = 0.05). The JD resulted in a significantly higher RCFA of 1.00 in the middle and apical thirds in both the straight and curved canals than the direct syringe method (p < 0.001). Voids were frequently observed in the direct syringe group, whereas JD showed no voids within the filled area. In the direct syringe groups, the apical area was more readily filled in both canal types when the tip was bound (p < 0.001). The JD was found to be more effective in filling the entire canal space than the binding and unbinding conditions of the direct injection method, particularly in the apical area.

Rare variants in long non-coding RNAs are associated with blood lipid levels in the TOPMed whole-genome sequencing study.

Long non-coding RNAs (lncRNAs) are known to perform important regulatory functions in lipid metabolism. Large-scale whole-genome sequencing (WGS) studies and new statistical methods for variant set tests now provide an opportunity to assess more associations between rare variants in lncRNA genes and complex traits across the genome. In this study, we used high-coverage WGS from 66,329 participants of diverse ancestries with measurement of blood lipids and lipoproteins (LDL-C, HDL-C, TC, and TG) in the National Heart, Lung, and Blood Institute (NHLBI) Trans-Omics for Precision Medicine (TOPMed) program to investigate the role of lncRNAs in lipid variability. We aggregated rare variants for 165,375 lncRNA genes based on their genomic locations and conducted rare-variant aggregate association tests using the STAAR (variant-set test for association using annotation information) framework. We performed STAAR conditional analysis adjusting for common variants in known lipid GWAS loci and rare-coding variants in nearby protein-coding genes. Our analyses revealed 83 rare lncRNA variant sets significantly associated with blood lipid levels, all of which were located in known lipid GWAS loci (in a ±500-kb window of a Global Lipids Genetics Consortium index variant). Notably, 61 out of 83 signals (73%) were conditionally independent of common regulatory variation and rare protein-coding variation at the same loci. We replicated 34 out of 61 (56%) conditionally independent associations using the independent UK Biobank WGS data. Our results expand the genetic architecture of blood lipids to rare variants in lncRNAs.

Improving radiotherapy in immunosuppressive microenvironments by targeting complement receptor C5aR1.

An immunosuppressive microenvironment causes poor tumor T cell infiltration and is associated with reduced patient overall survival in colorectal cancer. How to improve treatment responses in these tumors is still a challenge. Using an integrated screening approach to identify cancer-specific vulnerabilities, we identified complement receptor C5aR1 as a druggable target, which when inhibited improved radiotherapy, even in tumors displaying immunosuppressive features and poor CD8+ T cell infiltration. While C5aR1 is well-known for its role in the immune compartment, we found that C5aR1 is also robustly expressed on malignant epithelial cells, highlighting potential tumor cell-specific functions. C5aR1 targeting resulted in increased NF-κB-dependent apoptosis specifically in tumors and not normal tissues, indicating that, in malignant cells, C5aR1 primarily regulated cell fate. Collectively, these data revealed that increased complement gene expression is part of the stress response mounted by irradiated tumors and that targeting C5aR1 could improve radiotherapy, even in tumors displaying immunosuppressive features.

Malnutrition in heart failure with preserved ejection fraction.

BACKGROUND: Malnutrition may be an important geriatric condition in adults with heart failure with preserved ejection fraction (HFpEF), but studies on its prevalence and associated clinical outcomes are limited. The aim of this study was to determine if malnutrition is associated with short-term morbidity and mortality in ambulatory patients with HFpEF. METHODS: We examined 231 patients with a clinical diagnosis of HFpEF seen at two dedicated academic HFpEF programs (Weill Cornell Medicine and Michigan Medicine) from June 2018 to April 2022. Malnutrition was defined by Mini-Nutritional Assessment Short Form (MNA-SF) scores ≤11. The primary endpoint was a 6-month composite of all-cause mortality and all-cause hospitalization. A Cox proportional-hazard models was used to examine the association between malnutrition and the primary endpoint, adjusting for race, prior hospitalization history, and the validated Meta-Analysis Global Group in Chronic (MAGGIC) heart failure prognostic risk score. RESULTS: The median age of the cohort was 73 years (interquartile range 64-81). The most common comorbid conditions included hypertension (prevalence 81%), atrial fibrillation (43%), and obesity (63%). The prevalence of malnutrition was 42% (n = 97), and MNA-SF scores did not significantly correlate with body mass index (R = -0.02, p = 0.71). At the 6-month follow-up, 62 patients (26.8%) were hospitalized and four patients died (1.7%). In a fully-adjusted analysis, malnutrition was independently associated with the composite outcome of all-cause mortality and all-cause hospitalization (HR 1.94 [95% CI: 1.17-3.20], p = 0.01). CONCLUSION: Despite a high prevalence of obesity, two out of five ambulatory adults with HFpEF are malnourished. Malnutrition was independently associated with adverse outcomes at 6 months. Future work is necessary to develop interventions that can address malnutrition.