Surrogate Test Research Articles

Clinical utility of quantitative immunoassays and surrogate virus neutralization tests for predicting neutralizing activity against the SARS-CoV-2 Omicron BA.1 and BA.5 variants.

Developing new antibody assays for emerging SARS-CoV-2 variants is challenging. SARS-CoV-2 surrogate virus neutralization tests (sVNT) targeting Omicron BA.1 and BA.5 have been devised, but their performance needs to be validated in comparison with quantitative immunoassays. First, using 1749 PRNT-positive sera, we noticed that log-transformed optical density (OD) ratio of wild-type (WT) sVNT exhibited better titer-correlation with plaque reduction neutralization test (PRNT) than % inhibition value. Second, we tried 798 dilutional titration tests with 103 sera, but nonlinear correlation between OD ratio and antibody concentration limited titration of sVNT. Third, the titer-correlations of two sVNT kits for BA.1 and two quantitative immunoassays for WT were evaluated with BA.1 and BA.5 PRNT. All tested kits exhibited a linear correlation with PRNT titers, but the sVNT kits exhibited high false-negative rates (cPass-BA.1 kit, 45.4% for BA.1and 44.2% for BA.5; STANDARD F-BA.1 kit, 1.9% for BA.1and 2.2% for BA.5), while quantitative immunoassays showed 100% sensitivity. Linear mixed-effects model suggested superior titer-correlation with PRNT for quantitative immunoassays compared to sVNT kits. Taken together, the use of quantitative immunoassays for WT, rather than rapid development of new kits, would be practical for predicting neutralizing activities against emerging new variants.

Humoral Response Kinetics and Cross-Immunity in Hospitalized Patients with SARS-CoV-2 WT, Delta, or Omicron Infections: A Comparison between Vaccinated and Unvaccinated Cohorts.

With the ongoing evolution of severe acute respiratory virus-2 (SARS-CoV-2), the number of confirmed COVID-19 cases continues to rise. This study aims to investigate the impact of vaccination status, SARS-CoV-2 variants, and disease severity on the humoral immune response, including cross-neutralizing activity, in hospitalized COVID-19 patients. This retrospective cohort study involved 122 symptomatic COVID-19 patients hospitalized in a single center. Patients were categorized based on the causative specific SARS-CoV-2 variants (33 wild-type (WT), 54 Delta and 35 Omicron) and their vaccination history. Sequential samples were collected to assess binding antibody responses (anti-S/RBD and anti-N) and surrogate virus neutralization tests (sVNTs) against WT, Omicron BA.1, and BA.4/5. The vaccinated breakthrough infection group (V) exhibited higher levels of anti-S/RBD compared to the variant-matched unvaccinated groups (UVs). The Delta infection resulted in a more rapid production of anti-S/RBD levels compared to infections with WT or Omicron variants. Unvaccinated severe WT or Delta infections had higher anti-S/RBD levels compared to mild cases, but this was not the case with Omicron infection. In vaccinated patients, there was no difference in antibody levels between mild and severe infections. Both Delta (V) and Omicron (V) groups showed strong cross-neutralizing activity against WT and Omicron (BA.1 and BA.4/5), ranging from 79.3% to 97.0%. WT (UV) and Delta (UV) infections had reduced neutralizing activity against BA.1 (0.8% to 12.0%) and BA.4/5 (32.8% to 41.0%). Interestingly, patients who received vaccines based on the ancestral spike exhibited positive neutralizing activity against BA.4/5, even though none of the study participants had been exposed to BA.4/5 and it is antigenically more advanced. Our findings suggest that a previous vaccination enhanced the humoral immune response and broadened cross-neutralizing activity to SARS-CoV-2 variants in hospitalized COVID-19 patients.

Pre-Operative Coagulation Test Results Do Not Correlate with Self-Bleeding Assessment Tool (Self-BAT) Scores

Background: Elective surgical procedures are planned hemostatic challenges and laboratory coagulation testing is often the typical approach to pre-operative hemostatic assessments. This is unfortunate as the prothrombin time [PT] and activated partial thromboplastin time [aPTT] are of suboptimal predictive value for surgical bleeding risk and are have limited screening ability for inherited bleeding disorders. To the contrary, personal and family history of bleeding are important predictors of inherited bleeding disorders, and thus also likely predictive of perioperative bleeding risk. Quantitative bleeding assessment tools (BATs) have been developed to standardize the patient bleeding history, and a patient self-administered BAT (Self-BAT) has been validated for the diagnosis of type 1 von Willebrand disease. As part of a large, multicentre prospective cohort study to assess the accuracy of the Self-BAT in predicting peri-operative bleeding, we aimed to determine the correlation between results of coagulation assays and the Self-BAT score in patients undergoing elective surgery. Methods: We enrolled adult patients with no previously identified bleeding disorder who were undergoing general elective surgery between 2017 to 2018. We excluded patients with any prior history of therapeutic dose antithrombotic use, and those undergoing cardiac, vascular, emergency, ophthalmologic, or dental surgeries. Bloodwork was drawn at the pre-operative visit, and included aPTT, PT, von Willebrand factor (VWF) antigen, VWF GP1b (ristocetin calibrated) activity, factor VIII activity, platelet function assay (PFA)-100 closure times. The pre-operative Self-BAT and all laboratory tests were analyzed at the same site . We used Spearman's correlation coefficient to determine the relationship between results of laboratory testing and the Self-BAT scores. An abnormal Self-BAT score was defined as ≥6 for women and ≥4 for men. R v4.3.1 was used to perform the analysis. Results: A total of 368 were enrolled in the study, the mean age was 56 years old (standard deviation: 14.4) and 64.3% were women. A total of 12 individuals (3.3%) had a first degree relative with a bleeding disorder. The most common planned procedure was orthopedic (61%), followed by neurosurgery (14.4%), and general surgery (8.2%). The median Self-BAT score for women was 2 (interquartile range [IQR]: 1-4.3) and 1 (IQR: 0-2) for men. Of the 236 women and 131 men, 42 (17.8%) and 17 (13%) had abnormal Self-bat score. We found no significant correlations between the aPTT PT, Factor VIII, VWF panel nor PFA-100 and the Self-BAT score (Table 1). There was also no correlation found for sex stratified analyses. Conclusions: Our findings confirm that unselected ‘routine’ and specialized coagulation test results do not correlate with an individual's bleeding history. Thus, indiscriminate testing should not be used as surrogate screening tests for bleeding disorders in the pre-operative setting. Specialized coagulation testing should be reserved for when a hemostatic defect is suspected based on a positive personal or family bleeding history. We look forward to the results of the primary objective of the larger cohort study; enrollment is more than half completed to determine if the Self-BAT score itself is an accurate predictor of peri-operative bleeding.

Asciminib Is Effective Against ABL1 Gene Fusions in Acute Lymphoblastic Leukemia but Only When the ABL1 SH3 Domain Is Present

Background Ph-like acute lymphoblastic leukemia (ALL) is high risk and comprised of many kinase activating gene fusions including those involving the ABL1 gene ( ABL1-rearranged, ABL1r). ABL1r ALL is associated with poor relapse-free and overall survival and incidence increases with age. Tyrosine kinase inhibitors (TKIs) such as dasatinib effectively target BCR::ABL1+ ALL and are in clinical trials for ABL1r ALL. However, resistance to TKIs is often observed, and novel treatments are required. The allosteric inhibitor asciminib binds the myristate pocket of Bcr-Abl kinase and is currently in Phase III trials for treatment of BCR::ABL1+leukemias. We assessed asciminib efficacy in ABL1r ALL. Methods Ba/F3 cells were transduced with four ABL1 fusions observed in ALL and asciminib efficacy evaluated in AnnexinV/7-AAD cell death assays (LD 50). Specific regions of the ABL1 fusion partner were deleted by site-directed mutagenesis (SDM). STAT5/CRKL phosphorylation and hCD45 levels were investigated by flow cytometry. Cells from three NUP214::ABL1 ALL patients were injected into NSG mice to establish patient-derived xenograft (PDX) models. Once mice reached 5% hCD45+ cells in peripheral blood (PB), treatment was commenced: vehicle control, dasatinib, asciminib. At experimental endpoint (>50% hCD45+ cells in PB) mice were humanely killed and organs harvested for analysis. Results Transduced Ba/F3 cells demonstrated varying sensitivity to asciminib (Table 1). While asciminib was not efficacious against Ba/F3 RCSD1::ABL1 and SNX2::ABL1 cells, Ba/F3 RANBP2::ABL1 (Heatley et al 2021)and NUP214::ABL1 cells demonstrated LD 50 concentrations within the clinically achievable range (3 µM, p<0.0001; 0.8 µM, p=0.0238 compared with control, respectively). Following asciminib treatment, Ba/F3 NUP214::ABL1 cells demonstrated reduced phosphorylation of STAT5 (mean fluorescence intensity [MFI]=1820 to 790, 57% reduction) and CRKL (MFI=2069 to 951, 54% reduction), confirming asciminib inhibits kinase signalling. No phosphorylation of STAT5 or CRKL was observed in control cells (MFI=290 and 406, STAT5 and CRKL, respectively). Additionally, ex vivo assessment of ETV6::ABL1 patient blasts demonstrated reduced phosphorylation of STAT5 (MFI=3395 to 1788, 47% reduction) and CRKL (MFI=5039 to 3000, 40% reduction) following asciminib treatment. Interrogation of the different ABL1 fusion breakpoints (Figure 1) revealed fusions demonstrating asciminib sensitivity retained exons 2 or 3 of ABL1 (corresponding to complete/partial SH3 and complete SH2 domains). Conversely, gene fusions lacking exons 1-3 (truncating the SH3 domain and retaining a partial SH2 domain) were resistant to asciminib suggesting a critical region in the SH3 and/or SH2 domain/s required for asciminib efficacy. To further investigate this we systematically deleted regions of ABL1 exon 3 using the NUP214::ABL1 construct as the model system and generated transduced Ba/F3 cells expressing different SDM regions. Cell death assays indicated the SH3 domain is critical for asciminib efficacy (Table 1). Asciminib also demonstrated efficacy in pre-clinical mouse modelsof NUP214::ABL1 ALL (where ABL1 exon 3 and partial SH3 domain is retained). Asciminib treatment significantly increased survival outcomes compared with control mice (84 vs 46d, p=0.01) by a time comparable to that of dasatinib treatment (93 vs 46d, p=0.0046). Results were confirmed in two additional NUP214::ABL1 PDX models demonstrating asciminib's potential to treat NUP214::ABL1 patients. Asciminib treatment reduced spleen and liver weights to that of healthy mice and resolved the leukemic immunophenotype of PB and bone marrow cells. Conclusion ABL1r ALL is associated with high rates of treatment failure and relapse. For the first time, we demonstrate the efficacy of asciminib in this high risk disease. Our data support addition of asciminib to treatment regimens of patients with NUP214::ABL1 ALL, the most common ABL class gene fusion affecting up to 6% of B-ALL patients and up to 10% of T-ALL patients. We also identify a critical region of ABL1 required for asciminib efficacy in ABL1r ALL. We propose interrogation of ABL1r patient breakpoints to determine inclusion/exclusion of the ABL1 SH3 domain as a surrogate test to be used in conjunction with in vitro kinase sensitivity assays for predicting whether a patient is likely to respond to asciminib treatment.

Hybrid immunity from SARS-CoV-2 infection and mRNA BNT162b2 vaccine among Thai school-aged children

ObjectiveTo compare the immune response of hybrid immunity – arising from SARS-CoV-2 infection and mRNA BNT162b2 vaccination – to that of 2-doses of vaccine. MethodsIn a subanalysis of BNT162b2 vaccine trial in 5 to 11-year-old children, There were 179 children who had hybrid immunity compared with 134 children with solely 2-dose vaccine. The immunological outcome was a surrogate virus neutralization test (sVNT) against the Omicron strain, BA.1, (%inhibition). An sVNT level ≥68 % inhibition was considered as protective immune response. ResultsFrom February to April 2022, 179 children had COVID-19 natural infection resulting in hybrid immunity included: Group1;prior vaccination(n = 17), Group2;after the first dose(n = 61), and Group3;after the second dose(n = 97). The proportion of children with protective immune response was higher in Group 3 and Group 1 – 61.9 % and 58.8 %, compared to 36.1 % and 34.3 % in Group 2 and comparator group (2 doses of vaccine), respectively. The geometric mean % inhibition of sVNT was higher in Group 1 (68.5, 95 %CI 55.5–84.6) and Group 3 (63.5, 95 %CI 55.5–72.6), followed by comparator group (49.6, 95 %CI 44.8–54.9) and Group 2 (42.1, 95 %CI 34.6–51.3), p < 0.001. ConclusionsImmune response that arises from BNT162b2 vaccine after natural infection and infection after 2 doses of BNT162b2 was higher than infection after partially-vaccinated children.

Estimation of fibre architecture and scar in myocardial tissue using electrograms: An in-silico study

Atrial Fibrillation (AF) is characterized by disorganized electrical activity in the atria and is known to be sustained by the presence of regions of fibrosis (scars) or functional cellular remodelling, both of which may lead to areas of slow conduction. Estimating the effective conductivity of the myocardium and identifying regions of abnormal propagation is therefore crucial for the effective treatment of AF. We hypothesize that the spatial distribution of tissue conductivity can be directly inferred from an array of concurrently acquired contact electrograms (EGMs). We generate a dataset of simulated cardiac AP propagation using randomized scar distributions and a phenomenological cardiac model and calculate contact EGMs at various positions on the field. EGMs are enriched with noise extracted from biological data acquired in the lab. A deep neural network, based on a modified U-net architecture, is trained to estimate the location of the scar and quantify conductivity of the tissue with a Jaccard index of 91%. We adapt a wavelet-based surrogate testing analysis to confirm that the inferred conductivity distribution is an accurate representation of the ground truth input to the model. We find that the root mean square error (RMSE) between the ground truth and our predictions is significantly smaller (pval<0.01) than the RMSE between the ground truth and surrogate samples.

Use of a point-of-care test to rapidly assess levels of SARS-CoV-2 nasal neutralising antibodies in vaccinees and breakthrough infected individuals

Despite SARS-CoV-2 vaccines eliciting systemic neutralising antibodies (nAbs), breakthrough infections still regularly occur. Infection helps to generate mucosal immunity, possibly reducing disease transmission. Monitoring mucosal nAbs is predominantly restricted to lab-based assays, which have limited application to the public. In this multi-site study, we used lateral-flow surrogate neutralisation tests to measure mucosal and systemic nAbs in vaccinated and breakthrough infected individuals in Australia and Singapore. Using three lateral flow assays to detect SARS-CoV-2 nAbs, we demonstrated that nasal mucosal nAbs were present in 71.4 (95% CI 56.3–82.9%) to 85.7% (95% CI 71.8–93.7%) of individuals with breakthrough infection (positivity rate was dependent upon the type of test), whereas only 20.7 (95% CI 17.1–49.4%) to 34.5% (95% CI 19.8–52.7%) of vaccinated individuals without breakthrough infection had detectible nasal mucosal nAbs. Of the individuals with breakthrough infection, collective mucosal anti-S antibody detection in confirmatory assays was 92.9% (95% CI 80.3–98.2%) of samples, while 72.4% (95% CI 54.1–85.5%) of the vaccinated individuals who had not experienced a breakthrough infection were positive to anti-S antibody. All breakthrough infected individuals produced systemic anti-N antibodies; however, these antibodies were not detected in the nasal cavity. Mucosal immunity is likely to play a role in limiting the transmission of SARS-CoV-2 and lateral flow neutralisation tests provide a rapid readout of mucosal nAbs at the point-of-care.

Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2.

SARS-CoV-2 infects humans and a broad spectrum of animal species, such as pets, zoo animals, and nondomestic animals. Monitoring infection in animals is important in terms of the risk of interspecies transmission and the emergence of new viral variants. Economical, fast, efficient, and sensitive diagnostic tests are needed to analyze animal infection. Double-antigen sandwich ELISA has the advantage of being multispecies and can be used for detecting infections caused by pathogens that infect several animal hosts. This study aimed to develop a double-antigen sandwich ELISA using two SARS-CoV-2 proteins, N and RBD. We compared its performance, when using these proteins separately, with an indirect ELISA and with a surrogate virus neutralization test. Positive and negative controls from a cat population (n = 31) were evaluated to compare all of the tests. After confirming that double-antigen sandwich ELISA with both RBD and N proteins had the best performance (AUC= 88%), the cutoff was adjusted using positive and negative samples from cats, humans (n = 32) and guinea pigs (n = 3). The use of samples from tigers (n = 2) and rats (n = 51) showed good agreement with the results previously obtained using the microneutralization test. Additionally, a cohort of samples from dogs with unknown infection status was evaluated. These results show that using two SARS-CoV-2 proteins in the double-antigen sandwich ELISA increases its performance and turns it into a valuable assay with which to monitor previous infection caused by SARS-CoV-2 in different animal species.

Efficacy of octenidine- and chlorhexidine-based wash-mitts against Candida albicans and Candida auris – a comparative study

Management of outbreaks of the newly emerging pathogen Candida auris may include use of antimicrobial wash-mitts for decolonization. However, currently there is little clinical evidence to support the wide adoption of 'whole-body decolonization' as part of the protocol to effectively manage C.auris outbreaks. The aim of this study was to investigate the chemical tolerance of C.auris compared with the surrogate test organism Candida albicans as established in the European Standards (EN). Two commercially available antiseptic-impregnated wash-mitts based on either chlorhexidine digluconate (CHG) or octenidine dihydrochloride (OCT) were studied. Comparison of susceptibility of C.auris and C.albicans was investigated based on the standardized test protocol EN 13624. Experiments were conducted using the impregnation liquid squeezed from the wash-mitts at a contact time of 30s at different concentrations between 0.5% and 97% in the presence of low organic soiling. Yeasticidal efficacy according to EN 13624 was found for the OCT wash-mitts at 30s at ≥10% concentration with C.albicans. In comparison, reduction ≥4 log10 was found at a much lower concentration of ≥1% for both C.auris strains. For the CHG wash-mitts, efficacy against C.albicans was below 2 log10 reduction at 97% concentration within 30s. Efficacy against the two C.auris strains was around 3 log10 reduction. Both C.auris strains were found to be significantly more susceptible when compared with C.albicans. Data also demonstrate that not all antiseptic-impregnated wash-mitts are equally effective against C.auris with OCT having a higher efficacy compared with CHG.

Vaccine-induced SARS-CoV-2 antibody response: the comparability of S1-specific binding assays depends on epitope and isotype discrimination.

Quantification of the SARS-CoV-2-specific immune response by serological immunoassays is critical for the management of the COVID-19 pandemic. In particular, neutralizing antibody titers to the viral spike (S) protein have been proposed as a correlate of protection (CoP). The WHO established the First International Standard (WHO IS) for anti-SARS-CoV-2 immunoglobulin (Ig) (NIBSC 20/136) to harmonize binding assays with the same antigen specificity by assigning the same unitage in binding antibody units (BAU)/ml. In this study, we analyzed the S1-specific antibody response in a cohort of healthcare workers in Germany (n = 76) during a three-dose vaccination course over 8.5 months. Subjects received either heterologous or homologous prime-boost vaccination with ChAdOx1 nCoV-19 (AstraZeneca) and BNT162b2 (Pfizer-BioNTech) or three doses of BNT162b2. Antibodies were quantified using three anti-S1 binding assays (ELISA, ECLIA, and PETIA) harmonized to the WHO IS. Serum levels of neutralizing antibodies were determined using a surrogate virus neutralization test (sVNT). Binding assays were compared using Spearman's rank correlation and Passing-Bablok regression. All assays showed good correlation and similar antibody kinetics correlating with neutralizing potential. However, the assays show large proportional differences in BAU/ml. ECLIA and PETIA, which detect total antibodies against the receptor- binding domain (RBD) within the S1 subunit, interact similarly with the convalescent plasma-derived WHO IS but differently with vaccine serum, indicating a high sensitivity to the IgG/IgM/IgA ratio. All three binding assays allow monitoring of the antibody response in COVID-19-vaccinated individuals. However, the assay-specific differences hinder the definition of a common protective threshold in BAU/ml. Our results highlight the need for the thoughtful use of conversion factors and consideration of method-specific differences. To improve the management of future pandemics and harmonize total antibody assays, we should strive for reference material with a well-characterized Ig isotype composition.

DETECTION OF SARS-COV-2 NEUTRALIZING ANTIBODIES IN RETROPHARYNGEAL LYMPH NODE EXUDATES OF WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) FROM NEBRASKA, USA.

Disease surveillance testing for emerging zoonotic pathogens in wildlife is a key component in understanding the epidemiology of these agents and potential risk to human populations. Recent emergence of SARS-CoV-2 in humans, and subsequent detection of this virus in wildlife, highlights the need for developing new One Health surveillance strategies. We used lymph node exudate, a sample type that is routinely collected in hunter-harvested white-tailed deer (WTD, Odocoileus virginianus) for surveillance of chronic wasting disease, to assess anti-SARS-CoV-2 neutralizing antibodies. A total of 132 pairs of retropharyngeal lymph nodes collected from Nebraska WTD harvested in Nebraska, US, in 2019 (pre-SARS-CoV-2 pandemic) and 2021 (post-SARS-CoV-2 pandemic) were tested for SARS-CoV-2 with reverse transcription PCR. Thereafter, exudates obtained from these same lymph nodes were tested for SARS-CoV-2 neutralizing antibodies using a surrogate virus neutralization test. Neutralizing antibodies were detected in the exudates with high diagnostic specificity (100% at proposed cutoff of 40% inhibition). Application of this testing approach to samples collected for use in other disease surveillance activities may provide additional epidemiological data on SARS-CoV-2 exposure, and there is further potential to apply this sample type to detection of other pathogens of interest.

Impact of Tyrosine Kinase Inhibitors on the Immune Response to SARS-CoV-2 Vaccination in Patients with Non-Small Cell Lung Cancer.

Immune dysregulation and cancer treatment may affect SARS-CoV-2 vaccination protection. Antibody production by B-cells play a vital role in the control and clearance of the SARS-CoV-2 virus. This study prospectively explores B-cell seroconversion following SARS-CoV-2 immunization in healthy individuals and non-small cell lung cancer (NSCLC) patients undergoing oncological treatment. 92 NSCLC patients and 27 healthy individuals' blood samples were collected after receiving any COVID-19 vaccine. Serum and mononuclear cells were isolated, and a serum surrogate virus neutralization test kit evaluated SARS-CoV-2 antibodies. B-cell subpopulations on mononuclear cells were characterized by flow cytometry. Patients were compared based on vaccination specifications and target mutation oncological treatment. A higher percentage of healthy individuals developed more SARS-CoV-2 neutralizing antibodies than NSCLC patients (63% vs. 54.3%; p = 0.03). NSCLC patients receiving chemotherapy (CTX) or tyrosine kinase inhibitors (TKIs) developed antibodies in 45.2% and 53.7%, of cases, respectively, showing an impaired antibody generation. CTX patients exhibited trends towards lower median antibody production than TKIs (1.0, IQR 83 vs. 38.23, IQR 89.22; p = 0.069). Patients receiving immunotherapy did not generate antibodies. A sub-analysis revealed that those with ALK mutations exhibited non-significant trends towards higher antibody titers (63.02, IQR 76.58 vs. 21.78, IQR 93.5; p = 0.1742) and B-cells quantification (10.80, IQR 7.52 vs. 7.22, IQR 3.32; p = 0.1382) against the SARS-CoV-2 spike protein than EGFR patients; nonetheless, these differences were not statistically significant. This study shows that antibodies against SARS-CoV-2 may be impaired in patients with NSCLC secondary to EGFR-targeted TKIs compared to ALK-directed treatment.

Longitudinal Analysis of SARS-CoV-2-Specific Cellular and Humoral Immune Responses and Breakthrough Infection following BNT162b2/BNT162b2/BNT162b2 and ChAdOx1/ChAdOx1/BNT162b2 Vaccination: A Prospective Cohort in Naive Healthcare Workers.

Assessing immune responses post-SARS-CoV-2 vaccination is crucial for optimizing vaccine strategies. This prospective study aims to evaluate immune responses and breakthrough infection in 235 infection-naïve healthcare workers up to 13-15 months after initial vaccination in two vaccine groups (108 BNT/BNT/BNT and 127 ChAd/ChAd/BNT). Immune responses were assessed using the interferon-gamma enzyme-linked immunospot (ELISPOT) assay, total immunoglobulin, and neutralizing activity through surrogate virus neutralization test at nine different time points. Both groups exhibited peak responses one to two months after the second or third dose, followed by gradual declines over six months. Notably, the ChAd group exhibited a gradual increase in ELISPOT results, but their antibody levels declined more rapidly after reaching peak response compared to the BNT group. Six months after the third dose, both groups had substantial cellular responses, with superior humoral responses in the BNT group (p < 0.05). As many as 55 breakthrough infection participants displayed higher neutralization activities against Omicron variants, but similar cellular responses compared to 127 infection-naïve individuals, suggesting cross-immunity. Distinct neutralization classifications (<30%, >80% inhibition) correlated with different ELISPOT results. Our study reveals diverse immune response patterns based on vaccine strategies and breakthrough infections, emphasizing the importance of understanding these dynamics for optimized vaccination decisions.

Heparin-Loaded Composite Coatings on Porous Stent from Pure Magnesium for Biomedical Applications.

Challenges associated with drug-releasing stents used in percutaneous transluminal coronary angioplasty (PTCA) encompass allergic reactions, prolonged endothelial dysfunction, and delayed stent clotting. Although absorbable stents made from magnesium alloys seem promising, fast in vivo degradation and poor biocompatibility remain major challenges. In this study, zirconia (ZrO2) layers were used as the foundational coat, while calcium phosphate (CaP) served as the surface layer on unalloyed magnesium specimens. Consequently, the corrosion current density was decreased to 3.86, from 13.3 μA/cm2. Moreover, a heparin-controlled release mechanism was created by co-depositing CaP, gelatin (Gel), and heparin (Hep) on the specimens coated with CaP/ZrO2, thereby boosting magnesium's blood compatibility and prolonging the heparin-releasing time. Techniques like X-ray diffractometry (XRD), focused ion beam (FIB) system, toluidine blue testing, UV-visible spectrometry, field emission scanning electron microscopy (FESEM), and surrogate tests for endothelial cell viability were employed to examine the heparin-infused coatings. The drug content rose to 484.19 ± 19.26 μg/cm2 in multi-layered coatings (CaP-Gel-Hep/CaP-Hep/CaP/ZrO2) from 243.56 ± 55.18 μg/cm2 in a single layer (CaP-Hep), with the controlled release spanning beyond 28 days. Also, cellular viability assessments indicated enhanced biocompatibility of the coated samples relative to those without coatings. This suggests the potential of magnesium samples after coating ZrO2 and CaP with Gel as candidates for porous biodegradable stents or even scaffolds in biomedical applications.

An Improved Canonical Correlation Analysis for EEG Inter-Band Correlation Extraction.

(1) Background: Emotion recognition based on EEG signals is a rapidly growing and promising research field in affective computing. However, traditional methods have focused on single-channel features that reflect time-domain or frequency-domain information of the EEG, as well as bi-channel features that reveal channel-wise relationships across brain regions. Despite these efforts, the mechanism of mutual interactions between EEG rhythms under different emotional expressions remains largely unexplored. Currently, the primary form of information interaction between EEG rhythms is phase-amplitude coupling (PAC), which results in computational complexity and high computational cost. (2) Methods: To address this issue, we proposed a method of extracting inter-bands correlation (IBC) features via canonical correlation analysis (CCA) based on differential entropy (DE) features. This approach eliminates the need for surrogate testing and reduces computational complexity. (3) Results: Our experiments verified the effectiveness of IBC features through several tests, demonstrating that the more correlated features between EEG frequency bands contribute more to emotion classification accuracy. We then fused IBC features and traditional DE features at the decision level, which significantly improved the accuracy of emotion recognition on the SEED dataset and the local CUMULATE dataset compared to using a single feature alone. (4) Conclusions: These findings suggest that IBC features are a promising approach to promoting emotion recognition accuracy. By exploring the mutual interactions between EEG rhythms under different emotional expressions, our method can provide valuable insights into the underlying mechanisms of emotion processing and improve the performance of emotion recognition systems.

Dogs and Cats Are Less Susceptible to the Omicron Variant of Concern of SARS-CoV-2: A Field Study in Germany, 2021/2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a pandemic of unprecedented extent. Besides humans, a number of animal species can be infected; however, in some species, differing susceptibilities were observed depending on the virus variant. Here, we serologically investigated cats and dogs living in households with human COVID-19 patients. The study was conducted during the transition period from delta as the dominating variant of concern (VOC) to omicron (BA.1/BA.2) to investigate the frequency of virus transmission of both VOCs from infected owners to their pets. The animal sera were tested by surrogate virus neutralization tests (sVNT) using either the original receptor-binding domain (RBD), enabling the detection of antibodies against the delta variant, or an omicron-specific RBD. Of the 290 canine samples, 20 tested positive by sVNT, but there were marked differences between the sampling time and, related thereto, the virus variants the dogs had contact to. While in November 2021, infected owners led to 50% seropositive dogs (18/36), only 0.8% (2/254) of animals with household contacts to SARS-CoV-2 between December 2021 and April 2022 tested positive. In all cases, the positive reaction was recorded against the original RBD. For cats, a similar pattern was seen, as in November 2021, 38.1% (16/42) tested positive, and between December 2021 and March 2022, only 5.0% (10/199). The markedly reduced ratio of seropositive animals during the period of omicron circulation suggests a considerably lower susceptibility of dogs and cats to this VOC. To examine the effect of further omicron subvariants, sera taken in the second and third quarter of 2022 from randomly selected cats were investigated. 2.3% (11/372) tested seropositive, and all of them showed a stronger reaction against the original RBD, further supporting the assumption of a lower susceptibility of companion animals to the omicron VOC.

Validation of a SARS-CoV-2 Surrogate Neutralization Test Detecting Neutralizing Antibodies against the Major Variants of Concern.

SARS-CoV-2 infection and/or vaccination elicit a broad range of neutralizing antibody responses against the different variants of concern (VOC). We established a new variant-adapted surrogate virus neutralization test (sVNT) and assessed the neutralization activity against the ancestral B.1 (WT) and VOC Delta, Omicron BA.1, BA.2, and BA.5. Analytical performances were compared against the respective VOC to the reference virus neutralization test (VNT) and two CE-IVD labeled kits using three different cohorts collected during the COVID-19 waves. Correlation analyses showed moderate to strong correlation for Omicron sub-variants (Spearman's r = 0.7081 for BA.1, r = 0.7205 for BA.2, and r = 0.6042 for BA.5), and for WT (r = 0.8458) and Delta-sVNT (r = 0.8158), respectively. Comparison of the WT-sVNT performance with two CE-IVD kits, the "Icosagen SARS-CoV-2 Neutralizing Antibody ELISA kit" and the "Genscript cPass, kit" revealed an overall good correlation ranging from 0.8673 to -0.8773 and a midway profile between both commercial kits with 87.76% sensitivity and 90.48% clinical specificity. The BA.2-sVNT performance was similar to the BA.2 Genscript test. Finally, a correlation analysis revealed a strong association (r = 0.8583) between BA.5-sVNT and VNT sVNT using a double-vaccinated cohort (n = 100) and an Omicron-breakthrough infection cohort (n = 91). In conclusion, the sVNT allows for the efficient prediction of immune protection against the various VOCs.

Exploring potential of Marshall-RT as simple performance test to evaluate rutting resistance of asphalt mixtures

ABSTRACT This study explored a new simple monotonic Marshall rutting test (Marshall-RT) to evaluate the rutting resistance of asphalt mixtures. The Marshall-RT test setup combines the Marshall fixture (at the bottom) and the IDT fixture (at the top). The rutting resistance of asphalt mixtures was evaluated using peak load (Pmax) and work done for the deformation damage obtained from the load-displacement curve. Nine plant-produced asphalt mixtures having different types of binders (unmodified, crumb rubber modified binder (CRMB)), and polymer modified binder (PMB)) and gradations having different nominal maximum aggregate sizes (NMAS) were evaluated. The Marshall-RT test parameters (i.e. Pmax and W P m a x ) have shown a strong correlation with fundamental (i.e. Flow Number (FN)), simulative (i.e. Hamburg wheel tracking test (HWTT)), and simple monotonic (i.e. Ideal rutting test (IDEAL-RT)) rutting tests. Moreover, the Marshall-RT test has shown good repeatability, the highest differentiation potential, and consistent ranking with other rutting tests. Furthermore, for high traffic levels (i.e. 100 MSA (Million Standard Axle)), at 50°C preliminary minimum threshold criteria were proposed for Marshall-RT (Pmax and W P m a x ), and IDEAL-RT (RTIndex) as 5 kN, 14 kN-mm, and 90, respectively, to avoid premature rutting failure. Thus, the newly developed monotonic Marshall-RT test can also be used as a surrogate simple performance test to estimate the rutting potential of asphalt mixtures during the mix design and plant production phases as quality control (QC)/quality assurance (QA).

The seroprevalence of SARS-CoV-2 infection in Malaysia: 7 August to 11 October 2020.

From the beginning of the COVID-19 pandemic until mid-October 2020, Malaysia recorded ~15,000 confirmed cases. But there could be undiagnosed cases due mainly to asymptomatic infections. Seroprevalence studies can better quantify underlying infection from SARS-CoV-2 by identifying humoral antibodies against the virus. This study was the first to determine the prevalence of SARS-CoV-2 infection in Malaysia'sgeneral population, as well as the proportion of asymptomatic and undiagnosed infections. This cross-sectional seroprevalence study with a two-stage stratified random cluster sampling design included 5,131 representative community dwellers in Malaysia aged ≥1year. Data collection lasted from 7 August to 11 October 2020 involving venous blood sampling and interviews for history of COVID-19 symptoms and diagnosis. Previous SARS-CoV-2 infection was defined as screened positive using the Wantai SARS-CoV-2 Total Antibody enzyme-linked immunosorbent assay and confirmed positive using the GenScript SARS-CoV-2 surrogate Virus Neutralization Test. We performed a complex sampling design analysis, calculating sample weights considering probabilities of selection, non-response rate and post-stratification weight. The overall weighted prevalence of SARS-CoV-2 infection was 0.49% (95%CI 0.28-0.85) (N = 150,857). Among the estimated population with past infection, around 84.1% (95%CI 58.84-95.12) (N = 126 826) were asymptomatic, and 90.1% (95%CI 67.06-97.58) (N = 135 866) were undiagnosed. Our study revealed a low pre-variant and pre-vaccination seroprevalence of SARS-CoV-2 infection in Malaysia up to mid-October 2020, with a considerable proportion of asymptomatic and undiagnosed cases. This led to subsequent adoption of SARS-CoV-2 antigen rapid test kits to increase case detection rate and to reduce time to results and infection control measures.

B-071 Comparison of Humoral and Cellular Immune Responses Between ChAd-BNT Heterologous Vaccination and BNT-BNT Homologous Vaccination Following the third BNT Dose: A Prospective Cohort Study

Abstract Background The differential immune responses after two additional BNT162b2 (BNT) doses between ChAdOx1 nCoV-10 (ChAd)-primed and BNT-primed groups have not been elucidated. The aim of this study was to compare vaccine-induced humoral and cellular immune responses and evaluate breakthrough infection between the two vaccination strategies. Methods In 221 healthy subjects (111 in the ChAd group), longitudinal immune responses were monitored at 3, 4, and 6 months after the 2nd dose and 1, 3, and 6 months after the 3rd dose. Humoral immunity was measured by two fully automated chemiluminescent immunoassays (Elecsys and Abbott) and a surrogate virus neutralization test (sVNT). Cellular immunity was assessed by two interferon-γ (IFN-γ) release assays (QuantiFERON SARS-CoV-2 and Covi-FERON). Results After the 2nd dose of BNT vaccination, total antibody levels were higher in the ChAd group, but IgG antibody and sVNT results were higher in the BNT group. Following the 3rd dose vaccination, binding antibody titers were significantly elevated in both groups (ChAD-BNT; 15.4 to 17.8-fold, BNT-BNT; 22.2 to 24.6-fold), and the neutralizing capacity was increased by 1.3-fold in both cohorts. The ChAd-BNT group had lower omicron neutralization positivity than the BNT-BNT group (P = 0.001) at 6 months after the 3rd dose. Cellular responses to the spike antigen also showed 1.7 to 3.0-fold increases after the 3rd dose, which gradually declined to the levels equivalent to before the 3rd vaccination. The ChAd cohort tended to have higher IFN-γ level than the BNT cohort for 3–6 months after the 2nd and 3rd doses. The frequency of breakthrough infection was higher in the ChAd group (44.8%) than in the BNT group (28.1%) (P = 0.0219). Breakthrough infection induced increased humoral responses in both groups, and increase of cellular response was significant in the ChAd group. Conclusion Our study showed differential humoral and cellular immune responses between ChAd-BNT-BNT heterologous and BNT-BNT-BNT homologous vaccination cohorts. The occurrence of low antibody levels in the ChAd-primed cohort in the humoral immune response may be associated with an increased incidence of breakthrough infections. Further studies are needed on the benefits of enhanced cellular immunity in ChAd-primed cohorts.