Human Protection Research Articles

B-087 Sars-cov-2 Nt Chip test performance: a novel microarray-based enzymatic immunoassay for in vitro identification of neutralizing antibodies against emerging variants

Abstract Background The COVID-19 pandemic has created a need for reliable and adaptable diagnostic tools to detect exposure and neutralizing antibodies to emerging SARS-CoV-2 variants. To address this need, we evaluated the performance of the SARS-CoV-2 NT Chip Test, an ELISA-like immunoblot assay that incorporates a customizable panel of purified antigens from the variants of interest. This assay detects antibodies that neutralize the interaction between the receptor-binding domain (RBD) of the Wuhan strain and the Omicron BA1 variant with the cellular ACE-2 receptor and provides a marker of prior infection through the nucleocapsid protein. Methods We analyzed 73 clinical samples from six distinct groups: GROUP 1: Pre-pandemic samples (33); GROUP 2: Samples from children naive to Wuhan and infected with the Omicron variant (10); GROUP 3: Samples from individuals infected only with the original Wuhan strain (10); GROUP 4: Samples from individuals vaccinated and/or infected with the Wuhan strain and the Omicron variant (10); GROUP 5: Samples collected from individuals with 0-2 days after Omicron BA.1 infection (5) and GROUP 6: Samples collected from the same individuals 10 days after Omicron BA.1 infection (5). Evaluation utilized a proprietary non-numerical (qualitative) validation spreadsheet. Performance metrics included sensitivity, specificity, observed agreement (≥90%), and kappa index (≥0.70). Results were compared against the manufacturer's validation using the PRNT viral neutralization assay. Results Our comparative evaluation showed the following results for each parameter: 1) Wuhan neutralizing antibodies (sensitivity and specificity of 92%, agreement of 92%, and kappa index of 0.84); 2) Omicron neutralizing antibodies (sensitivity and specificity of 100%, agreement of 100%, and kappa index of 1.00); 3) Anti-nucleocapsid antibodies (sensitivity of 97%, specificity of 98%, agreement of 97%, and kappa index of 0.94). Three samples from GROUP 1 showed positive results for Wuhan RBD and one for Nucleocapsid, all close to the cutoff value. Four samples from GROUP 2 showed positive results for Wuhan RBD, which may be attributed to maternal antibodies. Three samples from GROUP 3 and three samples from GROUP 4, showed negative results for nucleocapsid, agreeing with the manufacturer's validation. Samples from GROUP 5 and GROUP 6 showed expected results, with Omicron RDB negative in GROUP 5, becoming positive in GROUP 6. Conclusions The test showed excellent performance in the verification of the three parameters in the populations studied, being approved in our laboratory. Due to its ability to incorporate antigens from emerging variants, it enables sensitive tracking of changes in neutralizing antibody profiles over time. The automated and customizable nature of the test provides an important tool for both public health epidemiological studies, monitoring changes in humoral immunity as this virus evolves, and for individual diagnostics, revealing the level of humoral protection of a patient against circulating strains in a timely manner to guide care. Although further studies are still warranted, the assay's ability to map evolving strain-specific serological immunity may offer valuable insights.

A Fluorescent Probe for Hydrazine Based on 4-hydroxycoumarin with High Selectivity and Sensitivity

Background: Therefore, the development of reliable analytical techniques with high selectivity and sensitivity to detect hydrazine is required for the protection of human health and safety. Objectives: Traditional methods for detecting N2H4 are frequently time-consuming, less accurate, and unsuitable for the analysis of living systems. Numerous fluorescent probes for hydrazine have been produced and gained some valuable results recently. The creation of a simple fluorescent probe for hydrazine detection is the goal of this project Method: In this study, 300 µL of probe 3-methyl-2-oxo-2H-chromen-7-yl propionate (MOCP) was mixed with an equivalent amount of the solution of each analyte to obtain the measurement solution. Following a 10-minute room temperature incubation period, the fluorescence spectra of the resultant solution were recorded. Results: The fluorescence intensity of the probe was noticeably enhanced when N2H4 was added to the probe, but almost no fluorescence enhancement was observed when other competitive ions were added. Conclusion: A hydrazine fluorescent probe based on 4-hydroxycoumarin fluorophore was developed. The probe MOCP displayed high sensitivity and selectivity for hydrazine, with a color change from colourless to blue for detection by the naked eye. Moreover, it demonstrated a low detection limit of 20 nM and a fast reaction time of 30 s.

Adjuvant-dependent impacts on vaccine-induced humoral responses and protection in preclinical models of nasal and genital colonization by pathogenic Neisseria.

Neisseria gonorrhoeae, which causes the sexually transmitted infection gonorrhea and N. meningitidis, a leading cause of bacterial meningitis and septicemia, are closely related human-restricted pathogens that inhabit distinct primary mucosal niches. While successful vaccines against invasive meningococcal disease have been available for decades, the rapid rise in antibiotic resistance has led to an urgent need to develop an effective gonococcal vaccine. Several surface antigens are shared among these two pathogens, making cross-species protection an exciting prospect. However, the type of vaccine-mediated immune response required to achieve protection against respiratory versus genital infection remains ill defined. In this study, we utilize well established mouse models of female lower genital tract colonization by N. gonorrhoeae and upper respiratory tract colonization by N. meningitidis to examine the performance of transferrin binding protein B (TbpB) vaccines formulated with immunologically distinct vaccine adjuvants. We demonstrate that vaccine-mediated protection is influenced by the choice of adjuvant, with Th1/2-balanced adjuvants performing optimally against N. gonorrhoeae, and both Th1/2-balanced and Th2-skewing adjuvants leading to a significant reduction in N. meningitidis burden. We further establish a lack of correlation between protection status and the humoral response or bactericidal titre. Combined, this work provides supports the feasibility for a single vaccine formulation to achieve pan-neisserial coverage.

Fc-engineered antibodies promote neutrophil-dependent control of Mycobacterium tuberculosis

Mounting evidence indicates that antibodies can contribute towards control of tuberculosis (TB). However, the underlying mechanisms of humoral immune protection and whether antibodies can be exploited in therapeutic strategies to combat TB are relatively understudied. Here we engineered the receptor-binding Fc (fragment crystallizable) region of an antibody recognizing the Mycobacterium tuberculosis (Mtb) capsule, to define antibody Fc-mediated mechanism(s) of Mtb restriction. We generated 52 Fc variants that either promote or inhibit specific antibody effector functions, rationally building antibodies with enhanced capacity to promote Mtb restriction in a human whole-blood model of infection. While there is likely no singular Fc profile that universally drives control of Mtb, here we found that several Fc-engineered antibodies drove Mtb restriction in a neutrophil-dependent manner. Single-cell RNA sequencing analysis showed that a restrictive Fc-engineered antibody promoted neutrophil survival and expression of cell-intrinsic antimicrobial programs. These data show the potential of Fc-engineered antibodies as therapeutics able to harness the protective functions of neutrophils to promote control of TB.

A guide to selecting high-performing antibodies for Protein-glutamine gamma-glutamyltransferase 2 (TGM2) for use in western blot, immunoprecipitation and immunofluorescence.

Protein-glutamine gamma-glutamyltransferase 2 (TGM2) is a Ca 2+ dependent enzyme that catalyzes transglutaminase cross-linking modifications. TGM2 is involved in various diseases, either in a protective or contributory manner, making it a crucial protein to study and determine its therapeutic potential. Identifying high-performing TGM2 antibodies would facilitate these investigations. Here we have characterized seventeen TGM2 commercial antibodies for western blot and sixteen for immunoprecipitation, and immunofluorescence. The implemented standardized experimental protocol is based on comparing read-outs in knockout cell lines against their isogenic parental controls. This study is part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While the use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.

Non-Abelian braiding of Fibonacci anyons with a superconducting processor

Quantum many-body systems with a non-Abelian topological order can host anyonic quasiparticles. It has been proposed that anyons could be used to encode and manipulate information in a topologically protected manner that is immune to local noise, with quantum gates performed by braiding and fusing anyons. Unfortunately, realizing non-Abelian topologically ordered states is challenging, and it was not until recently that the signatures of non-Abelian statistics were observed through digital quantum simulation approaches. However, not all forms of topological order can be used to realize universal quantum computation. Here we use a superconducting quantum processor to simulate non-Abelian topologically ordered states of the Fibonacci string-net model and demonstrate braidings of Fibonacci anyons featuring universal computational power. We demonstrate the non-trivial topological nature of the quantum states by measuring the topological entanglement entropy. In addition, we create two pairs of Fibonacci anyons and demonstrate their fusion rule and non-Abelian braiding statistics by applying unitary gates on the underlying physical qubits. Our results establish a digital approach to explore non-Abelian topological states and their associated braiding statistics with current noisy intermediate-scale quantum processors.

Manipulated Notions of Islam Regarding Women: An Analysis of The Breadwinner by Deborah Ellis under Islamic Feminism

This research attempts to examine the suffocated lives of women in Afghanistan in the novel, The Breadwinner by Deborah Ellis under the lens of Islamic Feminism presented by Fatema Mernissi in Islam and Democracy. Islam has given all the fundamental rights to women. Despite being under the protection of men, women ought to have their own individuality as both genders shall stand equal for their deeds on the day of judgement. This research identifies the manipulated notions of Islam regarding women in the respective novel through the framework of Islamic feminism. The theory emphasizes that Islam allows women to celebrate their femininity, but the misinterpretation of Quranic verses has misled Muslims into believing otherwise. Thus, the research contributes to the struggle of identifying Afghan women's rights within the outside and domestic sphere which have been demolished by men, with the aid of Islamic feminism. In The Breadwinner, Taliban’s agenda is to set a religious narrative that women are submissive and they should be confined inside the four walls. In contrast, the reality is entirely opposite as Islam provides equal rights to both sexes on human grounds.

Duration of Protection and Humoral Immune Response in Nile Tilapia (Oreochromis niloticus L.) Vaccinated against Streptococcus agalactiae.

Streptococcosis caused by Streptococcus agalactiae (S. agalactiae) is a major bacterial disease affecting the production of Nile tilapia (Oreochromis niloticus L.), causing significant economic losses due to mortality in the growing phase. Vaccination is the most effective method for preventing streptococcosis on Nile tilapia farms. In Brazil, the major tilapia-producing regions have long production cycles (6-10 months) and harvest tilapias weighing over 900 g for fillet production. Thus, data on the duration of the humoral immune response and protection in farmed tilapia have not been reported or are poorly described. Furthermore, the efficiency of serological testing for the long-term monitoring of immune responses induced by vaccination against S. agalactiae has never been addressed. This study evaluated the duration of protection and humoral immune response induced in Nile tilapia vaccinated against S. agalactiae until 300 days post-vaccination (dpv). The immunization trial was composed of two groups: vaccinated (Vac), vaccinated intraperitoneally with a commercial vaccine, and unvaccinated (NonVac) group, injected fish with sterile saline solution. At 15, 30, 150, 180, 210, and 300 dpv, blood sampling was conducted to detect anti-S. agalactiae IgM antibodies using indirect Enzyme-Linked Immunosorbent Assay (ELISA), and the fish were challenged with pathogenic S. agalactiae to determine the duration of vaccine protection through relative percentage survival (RPS). Spearman's rank correlation was performed between the ELISA optical density (OD) of vaccinated tilapia and the duration of vaccine protection (RPS). The mean cumulative mortality in NonVac and Vac groups ranged from 65 to 90% and less than 35%, respectively. The average RPS was 71, 93, 94, 70, 86, and 67% at 15, 30, 150, 180, 210, and 300 dpv, respectively. RPS revealed that the vaccine provided protection from 15 to 300 dpv. The specific anti-S. agalactiae IgM antibody levels were significantly higher in the Vac group than that non-Vac group up to 180 dpv. The vaccinated fish exhibited significant protection for up to 10 months after vaccination. There was a positive correlation between the antibody response and RPS. This study revealed that a single dose of commercial vaccine administered to Nile tilapia can confer long-term protection against S. agalactiae and that indirect ELISA can monitor the duration of the humoral immune response for up to six months following vaccination. Finally, vaccine protection over six months can be associated with other components of the fish immune system beyond the humoral immune response by IgM antibodies.

Efficient signal sequence of mRNA vaccines enhances the antigen expression to expand the immune protection against viral infection

The signal sequence played a crucial role in the efficacy of mRNA vaccines against virus pandemic by influencing antigen translation. However, limited research had been conducted to compare and analyze the specific mechanisms involved. In this study, a novel approach was introduced by substituting the signal sequence of the mRNA antigen to enhance its immune response. Computational simulations demonstrated that various signal peptides differed in their binding capacities with the signal recognition particle (SRP) 54 M subunit, which positively correlated with antigen translation efficiency. Our data revealed that the signal sequences of tPA and IL-6-modified receptor binding domain (RBD) mRNA vaccines sequentially led to higher antigen expression and elicited more robust humoral and cellular immune protection against the SARS-CoV-2 compared to the original signal sequence. By highlighting the importance of the signal sequence, this research provided a foundational and safe approach for ongoing modifications in signal sequence-antigen design, aiming to optimize the efficacy of mRNA vaccines.

High Transferability of Neutralizing Antibodies against SARS-CoV-2 to Umbilical Cord Blood in Pregnant Women Vaccinated with BNT162b2 XBB.1.5: A Retrospective Cohort Study.

Coronavirus disease 2019 (COVID-19) can lead to severe respiratory illness, rapid disease progression, and higher rates of intensive care unit admission in pregnant women. Infection during pregnancy is associated with an increased risk of preterm delivery, cesarean section, fetal dysfunction, preeclampsia, and perinatal death. Vertical transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from pregnant women to their fetuses has also been observed. Although severe infections in neonates and infants are rare, newborns can experience serious consequences from COVID-19 due to their suboptimal humoral immune system protection. The amino acids in the structural proteins of SARS-CoV-2 are constantly mutating. Since around January 2023, COVID-19, caused by omicron-type SARS-CoV-2 variants, has been prevalent globally. These variants can evade the immune response triggered by traditional mRNA-based COVID-19 vaccines, such as BNT162b2. Therefore, vaccination with BNT162b2 XBB.1.5, which provides protection against omicron-type SARS-CoV-2 variants, is recommended. This retrospective cohort study included 148 pregnant women who received the BNT162b2 XBB.1.5 vaccine at 30 partner medical institutions from September 2023 to January 2024. We examined the titers of anti-spike glycoprotein SARS-CoV-2 immunoglobin G (IgG) and IgA in the blood and umbilical cord blood obtained from the participants using ELISA. Anti-spike glycoprotein SARS-CoV-2 IgG and IgA titers were highest in the blood and cord blood at late gestational age (28-34 weeks). No serious side effects or adverse events were observed in either the pregnant women or their newborns. Pregnant women who received the BNT162b2 XBB.1.5 vaccine during gestational weeks 28 to 34 had the highest titers of anti-omicron SARS-CoV-2 variant antibodies in their blood. Moreover, these antibodies were transferred to their umbilical cord blood. To validate our findings, large cohort clinical studies involving numerous pregnant women are warranted. This study was funded by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) and Grants-in-Aid for Medical Research from the Japan Agency for Medical Research and Development (AMED).

A guide to selecting high-performing antibodies for Protein-glutamine gamma-glutamyltransferase 2 (TGM2) for use in western blot, immunoprecipitation and immunofluorescence

Protein-glutamine gamma-glutamyltransferase 2 (TGM2) is a Ca2+ dependent enzyme that catalyzes transglutaminase cross-linking modifications. TGM2 is involved in various diseases, either in a protective or contributory manner, making it a crucial protein to study and determine its therapeutic potential. Identifying high-performing TGM2 antibodies would facilitate these investigations. Here we have characterized seventeen TGM2 commercial antibodies for western blot and sixteen for immunoprecipitation, and immunofluorescence. The implemented standardized experimental protocol is based on comparing read-outs in knockout cell lines against their isogenic parental controls. This study is part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While the use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.

Secreted phospholipase PLA2G2E contributes to regulation of T cell immune response against influenza virus infection.

The involvement of secreted phospholipase A2s in respiratory diseases, such as asthma and respiratory viral infections, is well-established. However, the specific role of secreted phospholipase A2 group IIE (PLA2G2E) during influenza virus infection remains unexplored. Here, we investigated the role of PLA2G2E during H1N1 influenza virus infection using a targeted mouse model lacking Pla2g2e gene (Pla2g2e-/-). Our findings demonstrated that Pla2g2e-/- mice had significantly lower survival rates and higher viral loads in lungs compared to wild-type mice following influenza virus infection. While Pla2g2e-/- mice displayed comparable innate and humoral immune responses to influenza virus challenge, the animals showed impaired influenza-specific cellular immunity and reduced T cell-mediated cytotoxicity. This indicates that PLA2G2E is involved in regulating specific T cell responses during influenza virus infection. Furthermore, transgenic mice expressing the human PLA2G2E gene exhibited resistance to influenza virus infection along with enhanced influenza-specific cellular immunity and T cell-mediated cytotoxicity. Pla2g2e deficiency resulted in perturbation of lipid mediators in the lung and T cells, potentially contributing to its impact on the anti-influenza immune response. Taken together, these findings suggest that targeting PLA2G2E could hold potential as a therapeutic strategy for managing influenza virus infections.IMPORTANCEThe influenza virus is a highly transmissible respiratory pathogen that continues to pose a significant public health concern. It effectively evades humoral immune protection conferred by vaccines and natural infection due to its continuous viral evolution through the genetic processes of antigenic drift and shift. Recognition of conserved non-mutable viral epitopes by T cells may provide broad immunity against influenza virus. In this study, we have demonstrated that phospholipase A2 group IIE (PLA2G2E) plays a crucial role in protecting against influenza virus infection through the regulation of T cell responses, while not affecting innate and humoral immune responses. Targeting PLA2G2E could therefore represent a potential therapeutic strategy for managing influenza virus infection.

The crinoid Jimbacrinus bostocki from the Lower Permian Cundlego Formation near Gascoyne Junction, Western Australia

Invertebrate macrofossils are superbly preserved in slabs of calcareous silty sandstone exposed in the bed of the Gascoyne River just west of Gascoyne Junction, Western Australia. Based on regional mapping, these rocks have been assigned to the shallow marine Cundlego Formation, which has been dated to the Early Permian. Many of the slabs have preserved the cladid, inadunate crinoid Jimbacrinus bostocki in large numbers, with densities of 150 individuals or more per square metre. Articulated crowns with complete or partial stems attached are often haphazardly aggregated in pods. Details on the morphology of J. bostocki are provided, particularly on juveniles which have cirriferous stems. Adult stems lack any obvious holdfast and are acutely geniculate below the calyx before being preferentially directed linearly in the homocrinid (E–BC) symmetry plane in the E ray direction. Crowns are commonly preserved with their arms arranged in a protective manner, also referred to as the ‘trauma’ posture. It is inferred that such individuals were likely transported alive by storm induced turbidity currents from a feeding position on a marine shelf to deeper, possibly more anoxic, waters, often accumulating en masse in syndepositional lows such as swales. They were subsequently smothered by sediment and rapidly buried. These fossiliferous deposits are interpreted as an “obrution” Lagerstätte. Using taphonomic and functional morphology analysis, possible options for the feeding posture of J. bostocki are proposed. Because of its advanced arm morphology comprising mainly oblique muscular articulations between brachials, the question whether adults were motile, rather than sessile, is addressed. Robbert J Willink [robblink@ozemail.com.au], 11 Coral Sea Court, Sunshine Beach, Queensland 4567, Australia. Tom Kapitany [tomk@crystal-world.com] Crystal World, 13 Olive Road, Devon Meadows, Victoria 3977, Australia.

Comparing population-level humoral and cellular immunity to SARS-Cov-2 in Bangalore, India

Two types of immunity, humoral and cellular, offer protection against COVID. Humoral protection, contributed by circulating neutralizing antibodies, can provide immediate protection but decays more quickly than cellular immunity and can lose effectiveness in the face of mutation and drift in the SARS-CoV-2 spike protein. Therefore, population-level seroprevalence surveys used to estimate population-level immunity may underestimate the degree to which a population is protected against COVID. In early 2021, before India began its vaccination campaign, we tested for humoral and cellular immunity to SARS-Cov-2 in representative samples of slum and non-slum populations in Bangalore, India. We found that 29.7% of samples (unweighted) had IgG antibodies to the spike protein and 15.5% had neutralizing antibodies, but at up to 46% showed evidence of cellular immunity. We also find that prevalence of cellular immunity is significantly higher in slums than in non-slums. These findings suggest (1) that a significantly larger proportion of the population in Bangalore, India, had cellular immunity to SARS-CoV-2 than had humoral immunity, as measured by serological surveys, and (2) that low socio-economic status communities display higher frequency of cellular immunity, likely because of greater exposure to infection due to population density.

Elevated Serum Levels of SARS-CoV-2 Anti-Spike S1 RBD IgM, IgG, and IgA in Pfizer-BioNTech Vaccinated, COVID-19 Infected, and COVID-19 Infected after Pfizer-BioNTech Vaccinated Individuals after One Month

Background: While protective immunity against some viruses, such as coronaviruses, is relatively short-lived, healing from acute infections of many different viruses, such as those caused by yellow fever, polio, measles, and smallpox, can give cell-mediated and humoral immunity for a lifetime. The main element in the long-term prevention of reinfection by most viruses might be due to the specific antibodies generated by plasma cells. Aim: This study aims to estimate the levels of SARS-CoV-2 Anti-Spike S1 RBD IgM, IgA, and IgG in serum among people in Baghdad after one month of receiving the Pfizer-BioNTech vaccine, infected with the SARS-CoV-2 virus after vaccination and COVID-19 infected patients respectively. Methods: A total of 120 volunteers were enrolled in this study, which was conducted between the 1st of November 2022 and the 13th of January 2023, and they were divided into four groups, each group containing 30 individuals. The study groups were categorized after one month into vaccinated with the Pfizer-BioNTech vaccine (BNT162b2), infected with SARS-CoV-2 after Pfizer vaccination, COVID-19 patients, and control. Results: The study presented a significant difference where (P value < 0.05) in the serum levels of Anti-Spike S1 RBD IgM, IgA, and IgG for all groups compared to the control. Serum levels of S1 RBD IgM Anti-Spike of SARS-CoV-2 in all groups were significantly increased (P value >0.05) compared to each other. For Anti-Spike S1 RBD IgA and IgG, there was no significant difference (P value >0.05) between the COVID-19-infected patients group and those infected by SARS-CoV-2 after the Pfizer vaccination group. Positive correlations were found among Anti-Spike S1 RBD IgM, IgA, and IgG levels in the serum. Conclusions: Natural infection by SARS-CoV-2 or vaccination with Pfizer-BioNTech (BNT162b2) provides significant humoral Immunological protection.

Impact of Antenatal SARS-CoV-2 Exposure on SARS-CoV-2 Neutralization Potency.

A pregnancy booster dose significantly reduces the risk and severity of COVID-19, and it is widely recommended. A prospective cohort study was conducted to compare the transplacental passage of maternal antibodies from vaccination or infection during three trimesters against both the vaccine-targeted Wuhan strain and the Omicron strain of SARS-CoV-2. Maternal-infant dyads from vaccinated mothers were collected between 6 June 2022 and 20 September 2022. We analyzed 38 maternal-infant dyads from mothers who had been infected with COVID-19 and 37 from mothers without any previous infection. Pregnant women who received their last COVID-19 vaccine dose in the third trimester exhibited the highest anti-spike protein antibody levels and neutralizing potency against both the Wuhan strain and Omicron BA.2 variant in their maternal and cord plasma. Both second- and third-trimester vaccination could lead to a higher level of neutralization against the Wuhan and Omicron strains. COVID-19 infection had a negative effect on the transplacental transfer ratio of SARS-CoV-2 antibodies. A booster dose during the second or third trimester is encouraged for the maximum transplacental transfer of humoral protection against COVID-19 for infants.

Humoral profiles of toddlers and young children following SARS-CoV-2 mRNA vaccination

Although young children generally experience mild symptoms following infection with SARS-CoV-2, severe acute and long-term complications can occur. SARS-CoV-2 mRNA vaccines elicit robust immunoglobulin profiles in children ages 5 years and older, and in adults, corresponding with substantial protection against hospitalizations and severe disease. Whether similar immune responses and humoral protection can be observed in vaccinated infants and young children, who have a developing and vulnerable immune system, remains poorly understood. To study the impact of mRNA vaccination on the humoral immunity of infant, we use a system serology approach to comprehensively profile antibody responses in a cohort of children ages 6 months to 5 years who were vaccinated with the mRNA-1273 COVID-19 vaccine (25 μg). Responses are compared with vaccinated adults (100 μg), in addition to naturally infected toddlers and young children. Despite their lower vaccine dose, vaccinated toddlers elicit a functional antibody response as strong as adults, with higher antibody-dependent phagocytosis compared to adults, without report of side effects. Moreover, mRNA vaccination is associated with a higher IgG3-dependent humoral profile against SARS-CoV-2 compared to natural infection, supporting that mRNA vaccination is effective at eliciting a robust antibody response in toddlers and young children.

Privacy-Preserving Identification of Cancer Subtype-Specific Driver Genes Based on Multigenomics Data with Privatedriver.

Identifying cancer subtype-specific driver genes from a large number of irrelevant passengers is crucial for targeted therapy in cancer treatment. Recently, the rapid accumulation of large-scale cancer genomics data from multiple institutions has presented remarkable opportunities for identification of cancer subtype-specific driver genes. However, the insufficient subtype samples, privacy issues, and heterogenous of aberration events pose great challenges in precisely identifying cancer subtype-specific driver genes. To address this, we introduce privatedriver, the first model for identifying subtype-specific driver genes that integrates genomics data from multiple institutions in a data privacy-preserving collaboration manner. The process of identifying subtype-specific cancer driver genes using privatedriver involves the following two steps: genomics data integration and collaborative training. In the integration process, the aberration events from multiple genomics data sources are combined for each institution using the forward and backward propagation method of NetICS. In the collaborative training process, each institution utilizes the federated learning framework to upload encrypted model parameters instead of raw data of all institutions to train a global model by using the non-negative matrix factorization algorithm. We applied privatedriver on head and neck squamous cell and colon cancer from The Cancer Genome Atlas website and evaluated it with two benchmarks using macro-Fscore. The comparison analysis demonstrates that privatedriver achieves comparable results to centralized learning models and outperforms most other nonprivacy preserving models, all while ensuring the confidentiality of patient information. We also demonstrate that, for varying predicted driver gene distributions in subtype, our model fully considers the heterogeneity of subtype and identifies subtype-specific driver genes corresponding to the given prognosis and therapeutic effect. The success of privatedriver reveals the feasibility and effectiveness of identifying cancer subtype-specific driver genes in a data protection manner, providing new insights for future privacy-preserving driver gene identification studies.

THE EFFECT OF EXTRACTION METHODS ON THE DETERMINATION OF THE CONTENT OF POLYPHENOLS AND ANTI-OXIDANT ACTIVITIES OF ENRICHED EGG POWDER

Background. The utilization of phenolic compounds is limited by many factors, such as low solubility, low permeability, lability, rapid release, vulnerability towards environmental impacts and low bioavailability. To overcome these constraints, polyphenols can be closed within lipid-based and protein-based nanoparticles. In this study, nanocapsules derived from egg powder, enriched with chokeberry pomace extracts were examined. The content of antioxidant compounds (polyphenols, anthocyanins, phenylpropanoids, total phenolic compounds) and antioxidative potential (with the participation of radicals: DPPH•, ABTS•+ and FRAP assays) were determined to assess the influence of the extraction method (heat extraction, ultrasound assisted extraction, stirring extraction) and encapsulation process of the fruit extract. Results and conclusions. In the present study, the highest amounts of most examined compounds (regardless of the powder type) were achieved by heat extraction. The lowest values were obtained by stirring the solution at room temperature. The highest antioxidative potential towards radicals was assessed when ultrasounds were used as the extraction factor. Furthermore, the closure of fruit extracts in the protein shell appears to act in a protective manner on the antioxidant compounds present in the extract. The results show that the extraction method applied in the phase of preparing samples to the analysis is of great importance for determining the antioxidant compound content and antioxidant capacity.

The dynamics of the content of immunoglobulins in the blood serum of piglets according to the actions of immunostimulating agents

Inconsistency in housing and feeding conditions, production stress, a combination of environmental factors, and technological solutions reduce the protective properties of the animal body, which often leads to a decrease in their productive properties. The work aimed to investigate changes in indicators of the humoral protection factor, in particular, the quantitative composition of immunoglobulins M, G, and A in the body of young pigs under the influence of immunostimulating agents Globigen® Pig Doser and Globigen® Jump Start, which include specific Ig Y. To perform this task in the conditions of the private farm of FG PE "Glynyany Agro" of Lviv region, Zolochiv district, which uses a single-phase method of raising pigs, a study was conducted with the involvement of 36 piglets-analogues of crossbreeds of the Great White and Landrace breeds, which were formed into three groups of 12 animals each. Piglets of the control group were fed feed of a standard diet. The piglets of the first experimental group were orally administered 2 ml/g of Globigen®Pig Doser during the first three days after birth. Then, according to the technology, they consumed the feed prescribed in the diet. The piglets of the second experimental group were given Globigen®Jump Start, which was mixed with the feed of the basic ration in the amount of 2 kg per ton of feed and, starting from the age of 7 days, was fed to the piglets according to the technology. It was established that the use of immunostimulating agents was characterized by an increase in Ig G and Ig A in the blood serum of piglets in the most critical period of their lives. Thus, in the blood serum of 30-day-old piglets under the influence of Globigen® Pig Doser, with a high level of probability (Р < 0.01), an increase of this class of antibodies to the level of 13.47 mg/ml was detected, with the use of Globigen® Jump Start, this indicator reached the mark 12.51 mg/ml. Since this period, colostral immunity, represented by placental immunoglobulins of class G and colostrum immunoglobulins of class A, is already exhausted; we assume that the formation of the body's immune response of young pigs has begun. We believe that the increase in the content of class A immunoglobulins, most likely the secretory form of the piglets themselves, when using Globigen® Jump Start, which reached the mark of 1.48 mg/ml, indicates the formation of immune resistance mechanisms. The effectiveness, specificity, and mechanism of action of these agents is based on the ability of their components, particularly Ig Y, to bind to pathogens or their fragments, to neutralize and safely remove them from the body, which helps to increase the barrier function of the intestinal mucosa. The obtained research results confirm the practicality of using Globigen® Pig Doser and Globigen® Jump Start immunostimulants during single-phase rearing of piglets to increase the level of non-specific resistance and obtain healthy young animals with a high level of viability.