Search Results (467)

Urine-based immunoassay is a non-invasive method with demonstrated utility in detecting anti-SARS-CoV-2 antibodies in unvaccinated patients with COVID-19. To evaluate urine’s potential for serological surveys in a real-world setting, SARS-CoV-2 serology was performed on urine samples from vaccinated individuals, both with and without prior confirmed COVID-19. (1) Methods: An in-house indirect ELISA was used to measure antibodies against recombinant spike (S) and nucleocapsid (N) proteins of SARS-CoV-2 in urine and paired serum from 149 individuals vaccinated with Janssen AD26.COV2.S, an S protein-based COVID-19 vaccine. (2) Results: Anti-S and anti-N levels were higher in the urine and serum of participants with confirmed prior COVID-19 compared to those without prior infection. Urinary anti-S effectively distinguished vaccinated individuals with (AUC = 0.96) and without (AUC = 0.88) prior infection from negative controls (non-vaccinated, non-previously infected individuals) (p < 0.0001). Among vaccinated participants, urinary anti-S and anti-N identified prior infection, with AUC values of 0.73 (p < 0.0001) and 0.60 (p = 0.03), respectively, being recorded. (3) Conclusions: Findings indicate that urinary anti-SARS-CoV-2 antibodies reflect AD26.COV2.S vaccination and previous COVID-19. To further advance the methodology, studies with larger sample sizes and a greater diversity of COVID-19 vaccines are required. Full article

This paper develops a novel design formula to estimate the residual strength of corroded stiffened cylindrical structures. It extends a previously established ultimate strength formulation for intact cylinders by introducing a corrosion-induced strength reduction factor. The foundational formula considers failure mode interactions like yielding, local buckling, overall buckling, and stiffener tripping. This research utilizes recent experimental and numerical investigations on corroded ring-stiffened cylinder models. Experimental results validate the numerical analysis method, showing good agreement in collapse pressures (2–4% difference) and shapes. The validated numerical method is then subject to an extensive parametric study, systematically varying corrosion characteristics. Results indicate a clear relationship between corrosion volume and strength reduction, with overall buckling being more sensitive. Based on these comprehensive results, a new empirical strength reduction factor (${\rho }_c$) is derived as a function of the corrosion volume ratio ($V_{non}$). This factor is integrated into the existing ultimate strength formula, allowing direct residual strength estimation for corroded structures. The proposed formula is rigorously verified against experimental and numerical data, showing excellent agreement (mean 1.00, COV 5.86%). This research provides a practical, accurate design tool for assessing the integrity and service life of corroded stiffened cylindrical structures. Full article

Since 2020, the Gs/Gd H5N1 influenza virus (clade 2.3.4.4b) has established itself within wild bird populations across Asia, Europe, and the Americas, causing outbreaks in wild mammals, commercial poultry, and dairy farms. The impacts on the bird populations and the agricultural industry has been significant, requiring a One Health approach to enhanced surveillance in both humans and animals. To support pandemic preparedness efforts, we evaluated the Cepheid Xpert Xpress CoV-2/Flu/RSV plus kit and the BioFire Respiratory 2.1 Panel for their ability to detect the presence of non-seasonal, zoonotic influenza A viruses, including circulating H5N1 viruses from clade 2.3.4.4b. Both assays effectively detected the presence of influenza virus in clinically-contrived nasal swab and saliva specimens at low concentrations. The results generated using the Cepheid Xpert Xpress CoV-2/Flu/RSV plus kit and the BioFire Respiratory 2.1 Panel, in conjunction with clinical and epidemiological findings provide valuable diagnostic findings that can strengthen pandemic preparedness and surveillance initiatives. Full article

At least three betacoronaviruses have spilled over from bats to humans and caused severe diseases, highlighting the threat of zoonotic transmission. Thus, it is important to enhance surveillance capabilities by developing tools capable of detecting a broad spectrum of bat-borne betacoronaviruses. Three monoclonal antibodies (mAbs) targeting the nucleocapsid (N) protein were generated using recombinant N proteins from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). The cross-reactivities of these mAbs were evaluated against a panel of betacoronaviruses. Sandwich ELISAs (sELISAs) were subsequently developed to detect bat-borne betacoronaviruses that have high zoonotic potential. Among the mAbs, 7A7 demonstrated the broadest cross-reactivity, recognizing betacoronaviruses from the Sarbecovirus, Merbecovirus and Hibecovirus subgenera. The first sELISA, based on mAbs 7A7 and 6G10, successfully detected N protein in all clinical swab samples from COVID-19 patients with cycle threshold (Ct) values < 25, achieving 75% positivity overall (12/16). Using this as a reference, a second sELISA was established by pairing mAb 7A7 with mAb 8E2, which binds to multiple merbecoviruses. This assay detected the N protein of two merbecoviruses, namely the human MERS-CoV and bat-borne HKU5-CoV, at high sensitivity and has a limit of detection (LOD) that is comparable to the first sELISA used successfully to detect COVID-19 infection. These broadly reactive mAbs could be further developed into rapid antigen detection kits for surveillance in high-risk populations with close contact with wild bats to facilitate the early detection of potential zoonotic spillover events. Full article

Background/Objectives: Coronaviruses (CoVs) are a large family of respiratory viruses that cause respiratory illnesses ranging from mild colds to severe diseases such as severe acute respiratory syndrome and pandemics. The nasal cavity is a primary site for CoV entry and transmission. The study aimed to prepare a novel mucoadhesive emulgel specifically formulated with simple, safe, and cost-effective excipients to create a barrier on the nasal mucosa that impedes CoV infection. This formulation strategy was specifically designed to enable rapid and straightforward in vivo translation, addressing a critical gap in preventive measures against respiratory viruses. Methods: Three emulgels, containing macadamia oil, Carbopol and different percentages of hydroxypropyl methylcellulose (1, 1.2 and 1.5% (w/v), HPMC), were properly prepared and characterized for mucoadhesion, viscosity, and spreadability. The biological activity against SARS-CoV-2 was evaluated in vitro on infected epithelial cells. Results: The emulgel with 1.2% HPMC demonstrated optimal physicochemical properties (mucoadhesion: 342 ± 9 mN/cm²; viscosity: 1080 ± 83 cp; spreadability: 7.27 ± 0.06 cm) suitable for nasal application. Importantly, in vitro biological assays demonstrated that this emulgel significantly inhibits SARS-CoV-2 infection in epithelial cells, indicating an effective barrier to viral diffusion. Conclusions: By employing readily available, safe, and inexpensive excipients, this novel mucoadhesive emulgel offers a practical, scalable, and rapidly translatable nasal prophylactic approach to prevent early SARS-CoV-2 infection, addressing a critical unmet need in pandemic preparedness. Full article

Vaccination with ChAdOx1 nCoV-19 is an important countermeasure to fight the COVID-19 pandemic. This vaccine enhances human immunoprotection against SARS-CoV-2 by inducing an immune response against the SARS-CoV-2 S protein. However, the immune-related genes induced by vaccination remain to be identified. This study employs feature ranking algorithms, an incremental feature selection method, and classification algorithms to analyze transcriptomic data from an experimental group vaccinated with the ChAdOx1 nCoV-19 vaccine and a control group vaccinated with the MenACWY meningococcal vaccine. According to different time points, vaccination status, and SARS-CoV-2 infection status, the transcriptomic data was divided into five groups, including a pre-vaccination group, ChAdOx1-onset group, MenACWY-onset group, ChAdOx1-7D group, and MenACWY-7D group. Each group contained samples with 13,383 RNA features and 1662 small RNA features. The results identified key genes that could indicate the efficacy of the ChAdOx1 nCoV-19 vaccine, and a classifier was developed to classify samples into the above groups. Additionally, effective classification rules were established to distinguish between different vaccination statuses. It was found that subjects vaccinated with ChAdOx1 nCoV-19 vaccine and infected with SARS-CoV-2 were characterized by up-regulation of HIST1H3G expression and down-regulation of CASP10 expression. In addition, IGHG1, FOXM1, and CASP10 genes were strongly associated with ChAdOx1 nCoV-19 vaccine efficacy. Compared with previous omics-driven studies, the machine learning algorithms used in this study were able to analyze transcriptome data faster and more comprehensively to identify potential markers associated with vaccine effect and investigate ChAdOx1 nCoV-19 vaccine-induced gene expression changes. These observations contribute to an understanding of the immune protection and inflammatory responses induced by the ChAdOx1 nCoV-19 vaccine during symptomatic episodes and provide a rationale for improving vaccine efficacy. Full article

Objectives: The aim of our study is to evaluate whether texture analysis of 68Ga-DOTATOC PET/CT images can predict clinical outcome in patients with neuroendocrine tumors (NET). Methods: Forty-seven NET patients who had undergone 68Ga-DOTATOC PET/CT were studied. Primary tumors were localized in the gastroenteropancreatic (n = 35), bronchopulmonary (n = 8), and other (n = 4) districts. NET lesions were segmented using an automated contouring program and subjected to texture analysis, thus obtaining the conventional parameters SUVmax and SUVmean, volumetric parameters of the primary lesion, such as Receptor-Expressing Tumor Volume (RETV) and Total Lesion Receptor Expression (TLRE), volumetric parameters of the lesions in the whole-body, such as wbRETV and wbTLRE, and texture features such as Coefficient of Variation (CoV), HISTO Skewness, HISTO Kurtosis, HISTO Entropy-log₁₀, GLCM Entropy-log₁₀, GLCM Dissimilarity, and NGLDM Coarseness. Patients were subjected to a mean follow-up period of 17 months, and survival analysis was performed using the Kaplan–Meier method and log-rank tests. Results: Forty-seven primary lesions were analyzed. Survival analysis was performed, including clinical variables along with conventional, volumetric, and texture imaging features. At univariate analysis, overall survival (OS) was predicted by age (p = 0.0079), grading (p = 0.0130), SUVmax (p = 0.0017), SUVmean (p = 0.0011), CoV (p = 0.0037), HISTO Entropy-log₁₀ (p = 0.0039), GLCM Entropy-log₁₀ (p = 0.0044), and GLCM Dissimilarity (p = 0.0063). At multivariate analysis, only GLCM Entropy-log₁₀ was retained in the model (χ² = 7.7120, p = 0.0055). Kaplan–Meier curves showed that patients with GLCM Entropy-log₁₀ >1.28 had a significantly better OS than patients with GLCM Entropy-log₁₀ ≤1.28 (χ² = 10.6063, p = 0.0011). Conclusions: Texture analysis of 68Ga-DOTATOC PET/CT images, by revealing the heterogeneity of somatostatin receptor expression, can predict the clinical outcome of NET patients. Full article

Background/Objectives: The co-circulation of both influenza viruses and SARS-CoV-2 poses a significant health risk, especially for the elderly. While vaccination against both diseases remains an effective strategy to reduce the burden of symptomatic infections, the effect of administering COVID-19 mRNA and seasonal influenza vaccines (COV-Flu) on elicited antibody responses has not been explored. Methods: Participants between 18 and 90 years old were vaccinated with COVID-19 mRNA vaccines (n = 67), seasonal influenza vaccines (n = 130), or both (n = 201) within a three-month period between 2021 and 2024. Serum hemagglutination-inhibition (HAI) titers against influenza A (H1N1, H3N2) and B (Yamagata, Victoria) strains were measured from the COV-Flu participants or the participants vaccinated with influenza vaccines only (mono-Flu). SARS-CoV-2 neutralization assays were performed on sera collected from the COV-Flu participants and the participants receiving the mRNA vaccine only (mono-COVID-19). Results: The administration of influenza virus vaccines and COVID-19 mRNA vaccines within a three-month period significantly enhanced the post-vaccination HAI titers against both influenza A and B vaccine components, particularly in the elderly (65–90) participants. There were no significant differences in SARS-CoV-2 neutralization titers in COV-Flu participants compared to mono-COVID-19 participants. Conclusions: Vaccination with both the COVID-19 mRNA and influenza vaccines enhances influenza-specific HAI titers without compromising the neutralization titers elicited by COVID-19 mRNA vaccination against SARS-CoV-2, especially in the elderly. These findings indicate the potential benefits of this approach, particularly for older adults, by boosting influenza virus vaccine-induced serum HAI activity while maintaining COVID-19 protective immunity. Full article

Introduction: In mouse models of chronic lymphocytic leukemia (CLL), an effective anti-leukemia immune response was obtained by depleting a specific regulatory T-cell (Treg) subset. While STAT5 signaling could alter the homeostasis of naïve (nTreg) and activated (aTreg) subsets, which are capable of suppressing also CLL patients’ responses to microbial antigens, perturbed STAT3 signaling could drive CXCR5 expression in circulating T-follicular regulatory cells (Tfr) and their entry into the lymph node/tumor microenvironment. Materials and Methods: By using phospho-specific flow cytometry, we monitored STAT signaling/phosphorylation (pSTAT), in vitro responses to Sars-Cov2-antigen-specific stimulation, and circulating Treg subsets in relation to Binet stage and total tumor mass/tumor distribution (TTM/TD) scoring in 62 patients with CLL. Results: The percentage of CXCR5⁺ Treg significantly increased in patients with Binet stage B disease, and Tfr-like subsets were associated with higher TTM and lower TD. The pSTAT3 levels in CD4⁺ T-cells were only significantly increased in patients undergoing therapy. Lower nTreg percentages correlated with increased disease duration, and an increased aTreg/nTreg ratio correlated with SARS-CoV-2-antigen-induced STAT5 signaling responses. Conclusions: The results show increased amounts of circulating CXCR5⁺ Tfr-like subsets in patients with extensive lymph node involvement and augmented STAT3 signaling in patients on therapy. While STAT5 responses may drive nTreg differentiation into aTreg, nTreg decline is associated with increased disease duration. Full article

This research investigates the structural behavior of high-strength concrete beams reinforced with carbon fiber-reinforced polymer (CFRP) bars and varying percentages of recycled concrete aggregate (RCA). The study examined 15 reinforced concrete beams (200 × 250 × 2000 mm) constructed with different RCA proportions (0%, 25%, 50%, 75%, and 100%) and tested at three shear span-to-depth ratios ($a/d$ = 1.5, 2.5, and 3.5), addressing a critical knowledge gap in sustainable structural engineering. Specimens exhibited compressive strengths of 55–67 MPa and reached ultimate load capacities of up to 198.4 kN. Notably, beams with 75% RCA achieved 35.7% higher capacity than control specimens at $a/d$ = 1.5, challenging conventional expectations about RCA performance. Failure modes transitioned from shear-dominated at $a/d$ = 1.5 to flexure-dominated at $a/d$= 3.5, with optimal ductility indices (up to 2.75) observed at $a/d$ = 2.5. Statistical analysis revealed significant correlations between $a/d$ ratio and performance metrics, with a perfect parabolic relationship for the ductility index (R² = 1.0, $p<0.001$). Comparison with ACI 440.1R15 predictions showed generally conservative estimates (mean experimental-to-predicted ratio = 1.02, COV = 16.9%). The findings demonstrate that high-strength concrete can successfully incorporate substantial RCA quantities (up to 75%) without compromising performance when using CFRP reinforcement, potentially reducing virgin material consumption by approximately 33% for sustainable construction applications. Full article

Concrete-filled steel tube (CFST) columns reinforced with latticed steel angles (LSA), referred to as CFST-LSA columns, have been widely adopted in practical engineering. Understanding their mechanical behavior under eccentric loading is crucial for ensuring structural safety and performance in engineering applications. Previous experimental studies have demonstrated that the incorporation of steel angles substantially improves both the axial capacity and ductility of CFST-LSA columns. Existing methods for determining the eccentric bearing capacity of CFST-LSA columns primarily rely on the normalized N/N_u-M/M_u interaction curve. However, this approach involves a complex calculation procedure for evaluating the eccentric bearing capacity. To address this limitation, this study proposes a theoretical model based on the limit equilibrium method to predict the eccentric bearing capacity of CFST-LSA columns. The proposed model explicitly integrates fundamental geometric and material parameters, thereby enabling a more efficient and programmable calculation of the eccentric bearing capacity. Comparisons between the proposed model and experimental results show good agreement, with a tested-to-predicted eccentric resistance ratio of 1.085 and a coefficient of variation (COV) of 0.022. The proposed model can serve as a practical calculation method for eccentric loading of CFST-LSA columns, facilitating their application in high-rise buildings and long-span bridges. Full article

The Fragile X mental retardation type 1 gene (FMR1) contains a CGG triplet cluster of varied length (30 repeats on average) located in its 5′ UTR. In its premutated state (54–200 repeats), FMR1 contributes to the pathogenesis of premature ovarian insufficiency (POI). Its gray zone alleles (41–54 repeats) are supposed to impair the ovarian function as well. In the case of a CGG repeat length > 200, Fragile X syndrome occurs. Post-transcriptional expression of FMR1 is regulated by microRNAs. Although miR-323a-3p overexpression suppresses FMR1 in various tissues, this relationship has not been evaluated in the human ovary. Additionally, this microRNA targets SMADs, which are suggested regulators of ovarian cell proliferation, growth, and function. This study investigated how FMR1 allele lengths with CGG repeat numbers n < 55 (normal and gray zone genotypes) relate to miR-323a-3p expression and how they may impact associated SMAD expression in human granulosa cells. COV434 cells and patient-derived GCs were used to evaluate FMR1, miR-323a-3p, and BMP/SMAD-pathway member expression levels. Briefly, miR-323a-3p was significantly upregulated in GCs of the gray zone group compared to the normal allele group (p < 0.0001), while the FMR1 level did not vary. Furthermore, the gray zone group showed a significant upregulation of BMPR2, SMAD1, SMAD4, and SMAD9. In contrast, the miR-323a-3p transfection of COV434 cells significantly downregulated SMAD3, SMAD4, SMAD5, and SMAD9, while the FMR1 and SMAD1 levels remained stable. Our findings highlight a CGG repeat number-dependent upregulation of miR-323a-3p and an alteration of the BMP/SMAD pathway, suggesting that these changes happen and contribute to impaired ovarian function independently. Full article

The peptide TAT-I24, a fusion of the TAT peptide (amino acids 48–60) and the 9-mer peptide I24, has been previously shown to neutralize several double-stranded (ds) DNA viruses in vitro. We have now extended the testing to potentially sensitive RNA viruses and analyzed the antiviral effect of the peptide against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). In Vero E6 cells, TAT-I24 neutralized the human 2019-nCoV isolate (Wuhan variant) in a dose-dependent manner, while it was unable to neutralize two SARS-CoV-2 variants of concern, Delta and Omicron. Moreover, TAT-I24 could not significantly neutralize any of the SARS-CoV-2 variants in the human lung carcinoma cell line Calu-3, which provides an alternative entry route for SARS-CoV-2 by direct membrane fusion. Therefore, a possible dependence on virus uptake by endocytosis was investigated by exposing Vero E6 cells to chloroquine (CQ), an inhibitor of endosomal acidification. The Wuhan variant was highly sensitive to inhibition by CQ, an effect which was further enhanced by TAT-I24, while the Delta variant was less sensitive to inhibition by higher concentrations of CQ compared to the Wuhan variant. The microscopic analysis of COS-7 cells using a rhodamine-labeled TAT-I24 (Rho-TAT-I24) showed the endosomal localization of fluorescent TAT-I24 and co-localization with transfected GFP-Rab14 but not GFP-Rab5. As these proteins are found in distinct endosomal pathways, our results indicate that the virus entry pathway determines sensitivity to the peptide. Full article

Background/Objectives: Determining whether 3,4-dihydroxy-6-[¹⁸F]fluoro-L-phenylalanine positron emission tomography/computed tomography ([¹⁸F]F-DOPA PET/CT) data indicate brain metastasis progression (MP) or brain radionecrosis (RN) is challenging. The aim of this study was to present a method usable in the clinical setting without dedicated software that relies on less than five minutes of SiPM PET/CT data collected immediately after [¹⁸F]F-DOPA injection. Methods: We prospectively enrolled 15 patients with 19 lesions. Each acquisition was conducted in list mode (LM) for 25 min using a four-ring SiPM PET/CT system. We reconstructed three volumes from the LM file: acquisition duration of 120 s (V120), 270 s (V270), and 10 min for the standard clinical volume (Vclin). We measured each lesion’s maximum voxel activity (LS_max) and the corresponding mean activity with its standard deviation (CL_mean, CL_sd). We then calculated the LS_max/CL_mean ratio and the coefficient of variation (COV), defined as 100 × (CL_sd/CL_mean). Results: Lesions were classified as RN (n = 7) and MP (n = 12). For all volumes, LS_max/CL_mean differed significantly. The COV parameter exhibited significant differences in all comparisons except for V120 vs. V270. The best diagnostic performances were observed for V120 and V270, with an accuracy of 94.7%. The AUC values were 97.6% in both cases. Conclusions: A simple, static [¹⁸F]F-DOPA PET/CT acquisition, starting 1.5 min after injection and lasting less than five minutes, permitted reaching excellent diagnostic performance (100% sensitivity, 91.7% specificity, and 97.6% AUC) in discriminating between RN and MP. Full article

Background: Pre-existing adenovirus immunity restricts the utilization of adenovirus-vectored vaccines. The current study aims to conduct a pooled analysis of eight clinical trials to evaluate the influence of pre-existing Ad5 neutralizing antibodies on immunogenicity of Ad5-nCoV. Methods: The primary outcome indicator of this pooled analysis is the geometric mean titers (GMTs) of live SARS-CoV-2 NAbs against the wild-type strain on day 28 post-vaccination. Participants were divided into two cohorts: an adolescent cohort comprising individuals aged 6–17 years and an adult cohort with individuals aged 18 years and older. Within each cohort, individuals were further categorized into three subgroups based on their Ad5-nCoV vaccination schedules: one subgroup received a single intramuscular dose as the primary regimen (Ad5-IM-prime), another received an intramuscular dose as the heterologous prime-boost regimen (Ad5-IM-boost), and the last subgroup received an aerosolized dose as the heterologous prime-boost regimen (Ad5-IH-boost). Results: A total of 3512 participants were included in this pooled analysis. In the Ad5-IM-prime subgroup, there were 1001 adolescents and 1450 adults; in the Ad5-IM-boost subgroup, there were 65 adolescents and 396 adults; and in the Ad5-IH-boost subgroup, there were 207 adolescents and 393 adults. In the adult cohort, the GMTs of NAbs against wild-type SARS-CoV-2 on day 28 post-vaccination for the Ad5-IM-prime, Ad5-IM-boost, and Ad5-IH-boost subgroups were 35.6 (95% CI: 32.0, 39.7), 358.3 (95% CI: 267.6, 479.6), and 2414.1 (95% CI: 2006.9, 2904.0), respectively, with negative (less than 1:12) pre-existing NAb titers compared to 10.7 (95% CI: 9.1, 12.6), 116.9 (95% CI: 84.9, 161.1), and 762.7 (95% CI: 596.2, 975.8), respectively, with high (greater than 1:1000) pre-existing NAb titers. A similar trend was observed in the adolescent cohort, where pre-existing immunity was found to reduce the peak of live SARS-CoV-2 Nabs post-vaccination. Conclusions: Regardless of whether Ad5-nCoV is administered as a primary vaccination regimen or as a heterologous prime-boost strategy, a negative impact on immunogenicity can still be observed in the presence of high pre-existing immunity. However, when primary immunization is achieved with inactivated COVID-19 vaccines, aerosol inhalation can significantly enhance the immunogenicity of Ad5-nCoV compared to intramuscular injections of Ad5-nCoV as a booster. Full article