Humoral Research Articles

HLA-C Peptide Repertoires as Predictors of Clinical Response during Early SARS-CoV-2 Infection

The human leukocyte antigen (HLA) system plays a pivotal role in the immune response to viral infections, mediating the presentation of viral peptides to T cells and influencing both the strength and specificity of the host immune response. Variations in HLA genotypes across individuals lead to differences in susceptibility to viral infection and severity of illness. This study uses observations from the early phase of the COVID-19 pandemic to explore how specific HLA class I molecules affect clinical responses to SARS-CoV-2 infection. By analyzing paired high-resolution HLA types and viral genomic sequences from 60 patients, we assess the relationship between predicted HLA class I peptide binding repertoires and infection severity as measured by the sequential organ failure assessment score. This approach leverages functional convergence across HLA-C alleles to identify relationships that may otherwise be inaccessible due to allelic diversity and limitations in sample size. Surprisingly, our findings show that severely symptomatic infection in this cohort is associated with disproportionately abundant binding of SARS-CoV-2 structural and non-structural protein epitopes by patient HLA-C molecules. In addition, the extent of overlap between a given patient’s predicted HLA-C and HLA-A peptide binding repertoires correlates with worse prognoses in this cohort. The findings highlight immunologic mechanisms linking HLA-C molecules with the human response to viral pathogens that warrant further investigation.

Preschool-age children maintain a distinct memory CD4+ T cell and memory B cell response after SARS-CoV-2 infection.

The development of the human immune system lasts for several years after birth. The impact of this maturation phase on the quality of adaptive immunity and the acquisition of immunological memory after infection at a young age remains incompletely defined. Here, using an antigen-reactive T cell (ARTE) assay and multidimensional flow cytometry, we profiled circulating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-reactive CD3+CD4+CD154+ T cells in children and adults before infection, during infection, and 11 months after infection, stratifying children into separate age groups and adults according to disease severity. During SARS-CoV-2 infection, children younger than 5 years old displayed a lower antiviral CD4+ T cell response, whereas children older than 5 years and adults with mild disease had, quantitatively and phenotypically, comparable virus-reactive CD4+ T cell responses. Adults with severe disease mounted a response characterized by higher frequencies of virus-reactive proinflammatory and cytotoxic T cells. After SARS-CoV-2 infection, preschool-age children not only maintained neutralizing SARS-CoV-2-reactive antibodies postinfection comparable to adults but also had phenotypically distinct memory T cells displaying high inflammatory features and properties associated with migration toward inflamed sites. Moreover, preschool-age children had markedly fewer circulating virus-reactive memory B cells compared with the other cohorts. Collectively, our results reveal unique facets of antiviral immunity in humans at a young age and indicate that the maturation of adaptive responses against SARS-CoV-2 toward an adult-like profile occurs in a progressive manner.

Identification of antibody targets associated with lower HIV viral load and viremic control.

High HIV viral loads (VL) are associated with increased morbidity, mortality, and on-going transmission. HIV controllers maintain low VLs in the absence of antiretroviral therapy (ART). We previously used a massively multiplexed antibody profiling assay (VirScan) to compare antibody profiles in HIV controllers and persons living with HIV (PWH) who were virally suppressed on ART. In this report, we used VirScan to evaluate whether antibody reactivity to specific HIV targets and broad reactivity across the HIV genome was associated with VL and controller status 1-2 years after infection. Samples were obtained from participants who acquired HIV infection in a community-randomized trial in Africa that evaluated an integrated strategy for HIV prevention (HPTN 071 PopART). Controller status was determined using VL and antiretroviral (ARV) drug data obtained at the seroconversion visit and 1 year later. Viremic controllers had VLs <2,000 copies/mL at both visits; non-controllers had VLs >2,000 copies/mL at both visits. Both groups had no ARV drugs detected at either visit. VirScan testing was performed at the second HIV-positive visit (1-2 years after HIV infection). The study cohort included 13 viremic controllers and 64 non-controllers. We identified ten clusters of homologous peptides that had high levels of antibody reactivity (three in gag, three in env, two in integrase, one in protease, and one in vpu). Reactivity to 43 peptides (eight unique epitopes) in six of these clusters was associated with lower VL; reactivity to six of the eight epitopes was associated with HIV controller status. Higher aggregate antibody reactivity across the eight epitopes (more epitopes targeted, higher mean reactivity across all epitopes) and across the HIV genome was also associated with lower VL and controller status. We identified HIV antibody targets associated with lower VL and HIV controller status 1-2 years after infection. Robust aggregate responses to these targets and broad antibody reactivity across the HIV genome were also associated with lower VL and controller status. These findings provide novel insights into the relationship between humoral immunity and viral containment that could help inform the design of antibody-based approaches for reducing HIV VL.

Artificial intelligence caters to banks and customer needs

Banking customers do not think, speak, text, or communicate as financial institutions do. For humans, banking is as simple as “check balance,” “transfer,” “credit card payment,” and we prefer to talk in that language. When users contact a customer service representative, they want accurate answers without spending too much time over the phone or email. However, during these “new normal” times, the waiting time to reach a customer service rep has tremendously increased. Human agents cannot scale up with the ever-increasing demand to resolve customers’ queries. More often than not, agents feel burned out as they do not have enough tools at their disposal for delivering fast response time. With conversational artificial intelligence (AI) kicking in, the technology uses machine learning (ML) and natural language processing (NLP) to create a better customer experience. With the right set of programs and rules written, AI/ML tools can do much more than chatting and providing basic information to customers. For example, you could also update your contact and residential address, check your credit card balance, activate your card, etc. Chatbots are no longer “bots” just with pre defined answers to limited questions fed into them, with conversational AI and NLP- they are at par with human like responses and answer as accurately as a human would do. The businesses which are time zone independent find them more helpful.

Heterogeneity of SARS-CoV-2 immune responses after the nationwide Omicron wave in China.

It remains unclear how previous infections and vaccinations influenced and shaped heterogeneous immune responses against Omicron and its variants in diverse populations in China. After the national wave of Omicron in early 2023, we evaluated serum levels of neutralizing antibodies (nAbs) against Omicron (B.1.1.529) and its variants (BA.5, BF.7, and CH1.1) in 33 COVID-19 convalescents and 40 uninfected vaccinees, using vesicular stomatitis virus-based pseudovirus neutralizing assay. In addition, we followed 34 Delta convalescent patients to compare their immune responses against Omicron before (late 2021) and after the Omicron wave (early 2023). NAbs at the acute phase of the disease were investigated in 50 Omicron inpatients, including 24 vaccinated and 26 unvaccinated patients. Among them, nasal mucosal IgA levels were measured in 42 subjects. Compared to vaccination, breakthrough infections significantly increased the breadth and magnitude of serum nAbs and mucosal IgA levels against Omicron variants. Exposure to Omicron but not Delta elicited stronger pan-Omicron responses. In Omicron inpatients, nAbs continued to rise as vaccination doses increased. However, in both vaccinees and convalescents, a fourth dose vaccination did not elicit higher nAbs against Omicron. Furthermore, nAbs against Omicron variants lasted longer than nAbs against WT SARS-CoV-2. Breakthrough infections of Omicron variants elicited specific immune responses against Omicron compared to vaccination and Delta infection. Although repeated vaccination revealed limited impacts on serum nAbs, populations at high risk of hospitalization may still benefit from continued vaccination.IMPORTANCEThe study described the specific humoral immunity against Omicron and its variants (BA.5, BF.7, and CH1.1) in diverse populations, including Delta-positive convalescent patients, Omicron-infected patients with a previous or current confirmed Delta infection, Omicron-positive patients, and healthy controls. In addition, we followed Delta convalescents for 1 year to evaluate the effect of a booster vaccine, breakthrough infection, and reinfection. Nasal mucosal IgA levels against SARS-CoV-2 were also examined. The findings of this study demonstrated the varied responses of individuals in different states following the outbreak of Omicron, highlighting the potential advantages of ongoing immunization for groups that are more vulnerable and have a greater likelihood of being hospitalized.

A Comparative Analysis of Different Xenorhabdus Strains Reveals a Virulent Factor, Cyclic Pro-Phe, Using a Differential Expression Profile Analysis of Non-Ribosomal Peptide Synthetases

Entomopathogenic bacteria, classified into the genus Xenorhabdus, exhibit a dual lifestyle as mutualistic symbionts to Steinernema nematodes and as pathogens to a broad range of insects. Bacterial virulence depends on toxin proteins that induce toxemia and various immunosuppressive secondary metabolites that cause septicemia. Particularly, the immunosuppressive properties of Xenorhabdus bacteria determine the variability of their insecticidal activities. This study explored the role of peptide metabolites in virulence and its variation among six bacterial strains across three species: X. nematophila, X. bovienii, and X. hominickii. Initially, their virulence significantly varied against a susceptible lepidopteran host, Maruca vitrata, but showed less variation against a tolerant coleopteran host, Tenebrio molitor, with high median lethal bacterial doses. In M. vitrata, virulence was strongly correlated with bacterial growth rate and inhibitory activity against phospholipase A2. Secondly, the six strains differed in the compositions of their secreted secondary metabolites, analyzed by GC-MS following ethyl acetate extraction. Notably, there was significant variation in the production of di- or tetra-peptides. Highly virulent strains commonly produced the cyclic Pro-Phe (cPF). Thirdly, the expression of non-ribosomal peptide synthetase (NRPS) genes varied greatly among the strains. NRPS genes were minimally expressed in the tolerant T. molitor and highly expressed in the susceptible M. vitrata. In M. vitrata, specific NRPS genes were markedly expressed in the virulent strains. Finally, cPF demonstrated potent immunosuppressive activity against the cellular and humoral responses of M. vitrata. The addition of cPF significantly enhanced the virulence against the tolerant T. molitor. These findings suggest that immunosuppression is necessary for the pathogenicity of Xenorhabdus bacteria, wherein NRPS products play a critical role in suppressing immune-associated factors in target insects.

Clonal landscape of autoantibody-secreting plasmablasts in COVID-19 patients.

Whereas severe COVID-19 is often associated with elevated autoantibody titers, the underlying mechanism behind their generation has remained unclear. Here we report clonal composition and diversity of autoantibodies in humoral response to SARS-CoV-2. Immunoglobulin repertoire analysis and characterization of plasmablast-derived monoclonal antibodies uncovered clonal expansion of plasmablasts producing cardiolipin (CL)-reactive autoantibodies. Half of the expanded CL-reactive clones exhibited strong binding to SARS-CoV-2 antigens. One such clone, CoV1804, was reactive to both CL and viral nucleocapsid (N), and further showed anti-nucleolar activity in human cells. Notably, antibodies sharing genetic features with CoV1804 were identified in COVID-19 patient-derived immunoglobulins, thereby constituting a novel public antibody. These public autoantibodies had numerous mutations that unambiguously enhanced anti-N reactivity, when causing fluctuations in anti-CL reactivity along with the acquisition of additional self-reactivities, such as anti-nucleolar activity, in the progeny. Thus, potentially CL-reactive precursors may have developed multiple self-reactivities through clonal selection, expansion, and somatic hypermutation driven by viral antigens. Our results revealed the nature of autoantibody production during COVID-19 and provided novel insights into the origin of virus-induced autoantibodies.

Agnostic B cell selection approach identifies antibodies against K. pneumoniae that synergistically drive complement activation

Antibody-dependent complement activation plays a key role in the natural human immune response to infections. Currently, the understanding of which antibody-antigen combinations drive a potent complement response on bacteria is limited. Here, we develop an antigen-agnostic approach to stain and single-cell sort human IgG memory B cells recognizing intact bacterial cells, keeping surface antigens in their natural context. With this method we successfully identified 29 antibodies against K. pneumoniae, a dominant cause of hospital-acquired infections with increasing antibiotic resistance. Combining genetic tools and functional analyses, we reveal that the capacity of antibodies to activate complement on K. pneumoniae critically depends on their antigenic target. Furthermore, we find that antibody combinations can synergistically activate complement on K. pneumoniae by strengthening each other’s binding in an Fc-independent manner. Understanding the molecular basis of effective complement activation by antibody combinations to mimic a polyclonal response could accelerate the development of antibody-based therapies against problematic infections.

Carbohydrate-Lectin Interactions Reprogram Dendritic Cells to Promote Type 1 Anti-Tumor Immunity.

Cancer vaccine development is inhibited by a lack of strategies for directing dendritic cell (DC) induction of effective tumor-specific cellular immunity. Pathogen engagement of DC lectins and toll-like receptors (TLRs) is thought to shape immunity by directing T cell function. Controlling downstream responses, however, remains a major challenge. A critical goal in advancing vaccine development involves the identification of receptors that drive type 1 cellular immunity. The immune system monitors cells for aberrant glycosylation (a sign of a foreign entity), but potent activation occurs when a second signal, such as single-stranded RNA or lipopolysaccharide, is present to activate TLR signaling. To exploit dual signaling, we engineered a glycan-costumed virus-like particle (VLP) vaccine that displays a DC-SIGN-selective aryl mannose ligand and encapsulates TLR7 agonists. These VLPs deliver programmable peptide antigens to induce robust DC activation and type 1 cellular immunity. In contrast, VLPs lacking this critical DC-SIGN ligand promoted DC-mediated humoral immunity, offering limited tumor control. Vaccination with glycan-costumed VLPs generated tumor antigen-specific Th1 CD4+ and CD8+ T cells that infiltrated solid tumors, significantly inhibiting tumor growth in a murine melanoma model. The tailored VLPs also afforded protection against the reintroduction of tumor cells. Thus, DC lectin-driven immune reprogramming, combined with the modular programmability of VLP platforms, provides a promising framework for directing cellular immunity to advance cancer immunotherapies and vaccines.

Abstract B055: Harnessing cytomegalovirus immunity against pancreatic tumors for immunotherapy

Abstract Immunotherapy has had very limited success in pancreatic cancer, due to its low mutational burden and immunosuppressive microenvironment. Our approach consists in redirecting pre- existing antiviral immunity against pancreatic tumors by delivering viral antigens using the tumor penetrating peptide iRGD .This peptide, targets the tumor vasculature through av integrins and neuropilin-1, delivering conjugated or co-administered cargo to tumors. Here we used mouse cytomegalovirus (CMV) as an infection model. CMV is a β-herpesvirus that induces a strong T-cell response in mice and humans, comprising &amp;gt;10% of all circulating CD4 and CD8 T cells mostly with effector-memory phenotype. Importantly, human CMV infects over 60% of the world’s population rendering it a suitable candidate for translation. Methods: Mice latently infected with CMV were orthotopically implanted with KPC pancreatic tumor cells and treated with systemic injections of vehicle or iRGD plus CMV class I and II binding peptides. Another cohort of age matched uninfected mice was used as a control. Tumor growth and mouse weight were monitored twice a week and CMV specific immune responses were measured in spleen, liver and tumor by flow cytometry, using tetramer staining and CMV specific peptide recall. Results: CMV infected mice receiving iRGD+CMV peptides responded to treatment as evidenced by delayed tumor growth associated with increased tumor necrosis and T cell infiltration. In addition, flow cytometry analysis of tumor, liver and spleen showed a significant increase in CMV specific T cells preferentially in the tumor. Surprisingly, these T cells preferentially infiltrated tumors regardless of the use of iRGD for tumor targeting of CMV peptides. Survival studies showed a marked increase in median survival, reaching 42 days in the infected and treated group, compared to 25 days in the vehicle treated one. The survival benefit was very similar in the presence or absence of iRGD. Combining this strategy with anti- PD1 and or anti-IL10R did not significantly improve the benefits of CMV therapy alone. On the other hand, combination with low dose gemcitabine (5mg/kg) significantly improved tumor growth delay and survival in the combination compared to either therapy alone. The median survival reached 49 days in the combination, compared to 39 for gemcitabine or CMV therapy alone and 32 for vehicle.Conclusions: We have recently performed single cell RNA sequencing in tumors from infected and CMV peptide or vehicle treated mice and will analyze the transcriptional profile and TCR sequence of the tumor infiltrating T cells. Preliminary analysis of this data shows a massive T cell infiltration and an increase in dendritic cells and macrophages in infected and treated mice. Taken together, this data shows we can deliver antigens to pancreatic tumors and elicit an anti-tumor immune response that results in delayed tumor growth and has a survival benefit. This is a mutation agnostic approach with high translational value, since more than 60% of the population has previous CMV immunity. Citation Format: Remi Marrocco, Jay Patel, Rithika Medari, Siming Sun, Kevin Gulay, Alexei Martsinkovskiy, Simon Brunel, Eduardo Lucero-Meza, Evangeline Mose, Andrew Lowy, Chris Benedict, Tatiana Hurtado de Mendoza. Harnessing cytomegalovirus immunity against pancreatic tumors for immunotherapy [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B055.

PSI-15 Validation of a rapid test to evaluate humoral immunity in Labrador Retrievers across multiple life stages

Abstract This study sought to validate the D2Dx rapid immunity test in canines and investigate the age-related changes in the humoral immune system of healthy large breed dogs. Blood plasma with EDTA anticoagulant was collected from healthy Labrador Retrievers (n = 80) housed at Four Rivers Kennel under IACUC protocol FRK-56. Age groups were defined as follows: puppy 0-1.5 yr, adults 1.5-7 yr, senior 7-10, and geriatric 10-14. Immunity biomarkers including c-reactive protein (CRP), interleukin 4 (IL-4), interferon gamma (IFN-γ), complement protein 3 (C3), and immunoglobulin G (IgG) were determined through canine specific enzyme-linked immunosorbent assay (ELISA) kits. The nanoparticle based D2Dx test, which evaluates the cumulative humoral immune response, was also performed. Data were analyzed using a one-way ANOVA between age groups, with a Tukey’s post hoc, and correlations between variables were evaluated through multivariate analysis. The D2Dx score was correlated with body weight, age, IgG, and C3 validating that the test was able to adequately measure the humoral immunity in canines (r ≥ 0.49). D2Dx score was significantly different between age groups (P &amp;lt; 0.01) with puppies having significantly reduced scores compared with all other groups (P &amp;lt; 0.01). C3 was significantly different between age groups (P &amp;lt; 0.01), with puppies significantly less than other life stages (P ≤ 0.03). The C3 of adults averaged significantly greater than seniors and trended greater than geriatric subjects (P ≤ 0.01, 0.07). IgG between ages was also significantly different, with puppies significantly less than other age groups (P &amp;lt; 0.01). There was no significant difference between age groups for IFN-γ, IL-4 ratio, IFN-γ, IL-4, or CRP (P ≥ 0.18). This study showed that the D2Dx Rapid Immunity Test is able to detect changes to humoral immunity in canines. Beyond evaluating the effectiveness of the D2Dx test, this study also provides important information about the under researched topic of canine humoral immunity development and senescence in a robust, healthy cohort of large breed dogs.

PSLBII-17 Supplementation with organic versus inorganic selenium during pregnancy to support ewe and lamb health

Abstract Selenium (Se) is essential for optimal ovine health and production, and sheep raised on Se-deficient soils can be vulnerable to Se deficiencies that can affect immune function. Organic or inorganic forms of Se supplementation can remedy these consequences. While inorganic Se is less costly, organic Se is more bioavailable and may be more efficient in providing Se to ewes and lambs. The objectives of this study were to determine how organic versus inorganic Se supplementation affects ewe Se status, Se transfer from ewes to lambs, and the antibody-mediated immune response and dermal hypersensitivity response (DHR) of ewes and their lambs. Dorset-Rideau ewes (n = 100) were enrolled in a randomized complete block design study. From gestation d (gd) 110 to postpartum d (ppd) 50, ewes were fed either a Se-deficient diet and supplemented with no Se, 0.3 mg inorganic Se, or 0.3 mg or 0.6 mg organic Se. Sera were collected on gd 110, 130, 140, and ppd 10 and 50 to determine ewe Se status. On gd 110 and 120, ewes received an intramuscular injection of 0.5 mg ovalbumin (OVA) antigen, and on gd 130 the DHR to OVA was assessed by measuring change in skinfold thickness 0, 6, 24 and 48 h post intradermal injection. To measure the transfer of Se from ewe to lamb, a subset of lambs was euthanized at ppd 10, and longissimus dorsi muscle samples were collected and analyzed for Se content. On ppd 20 and 30, lambs received an intramuscular injection of 0.5 mg hen egg white lysozyme (HEWL) antigen, and on ppd 40 the DHR to HEWL was assessed using the aforementioned protocol. Analysis of ewe sera showed a positive correlation between organic Se supplementation and greater Se concentrations (P &amp;lt; 0.01). Moreover, statistical analyses of the DHR results using a generalized least squares linear model (RStudio) indicate that Se supplementation increased the immune response compared with the non-supplemented ewes (P &amp;lt; 0.01). However, no significant differences between organic and inorganic Se supplementation were found. In lambs, greater ewe Se supplementation resulted in greater Se muscle content (P &amp;gt; 0.01). and organic Se supplementation to ewes significantly increased lamb Se muscle content compared with inorganic Se supplementation (P &amp;gt; 0.01). Finally, analyses of the lamb DHR showed that maternal organic Se supplementation improved the lamb immune response compared with the control group (P &amp;gt; 0.05). Therefore, organic Se supplementation optimally enhances ewe and lamb Se status and their adaptive immune response compared with the inorganic and control groups. This knowledge will guide optimal ewe Se-supplementation choices during pregnancy and postpartum, thus promoting lamb and ewe health.

PSIII-27 Blood biochemistry, histology, and metabolomic profiling of contaminated with mycotoxin: A repeated dose oral toxicity study in rats

Abstract Mycotoxins causes diseases and physiological responses in humans and livestock. This study was performed to estimate the effects of binary mixtures mycotoxin {e.g., deoxynivalenol (DON) and fumonisin (FB)] on blood biochemistry, histology, and metabolic profiles in rats fed with different toxin concentrations. Experimental animals were administered by oral gavage with 0.9% saline, DON (1, 5, and 10 mg/kg BW), FB (1, 5, and 10 mg/kg) at 8 wk of age. Histological changes, metabolomic profiling, and blood biochemical analysis were performed Masson’s trichrome stain kit, liquid chromatography mass spectrometry, and VetTest chemistry analyzer, respectively. The final BW of the rats contaminated with mixtures mycotoxin significantly decreased compared with the control group. Biochemistry results and fibrosis were also significantly differentially regulated according to mycotoxin concentrations. Additionally, we observed collagen fibers in the liver and kidney showed blue staining after toxin contamination. For metabolomic profiling, data were analyzed using MetaboAnalyst 4.0. Results were analyzed by multivariate analysis to elucidate the potential compounds. The toxin-treated groups observed discriminating metabolites in the liver and kidney tissue but no differences in ileum. The profiling revealed potential metabolites such as glycine, serine and threonine metabolism. Phosphatidylethanolamine and tryptophan were identified with high sensitivity as candidate biomarkers via variable importance in the projection plot in piglets fed with mycotoxin (FDR &amp;lt; 0.05; P &amp;lt; 0.05). The differently treated groups showed discriminating metabolites in different concentrations. In conclusion, these findings suggest that mycotoxin exposure disturbs histological changes and metabolites. Dietary toxins in rats may better understand the physiological effects consistent with metabolism, which are commonly reported in animals.

Impacts of dietary supplementation of chitosan nanoparticles on growth, carcass traits nutrient digestibility, blood biochemistry, intestinal microbial load and meat quality of broilers

Abstract This study explores the impact of chitosan nanoparticles (CNP) on the performance, nutrient digestibility, blood biochemical, immunity, microbial load, carcass traits and meat attributes of broilers. A total of 200 7-day-old Cobb chicks were distributed to four groups, each replicated five times, with ten birds in each replicate. The experimental diets were as follows: First group fed basal diet only (control); 2nd, 3rd and 4th groups received a basal diet supplemented with 0.2, 0.3 and 0.4 g CNP/kg of feed, respectively. Results showed that the body weight (BW) and body weight gain (BWG) significantly improved (P &amp;lt; 0.05) in the birds belonging to the 0.4 CNP group compared to the other groups. The best feed efficiency (FCR) was found in the group supplemented with a 0.4 g CNP/kg diet. The digestibility coefficients for dry matter (DM), and crude protein (CP) were significantly higher, and ether extract (EE) was significantly lower in the 0.4 g CNP/kg group than in other groups (P&amp;lt; 0.05). Broiler birds of the 0.4 CNP group had significantly (P&amp;lt; 0.05) reduced serum cholesterol, AST, and ALT levels. The humoral immunity (increased serum IgG and IgM levels) tended to improve in birds fed 0.3 and 0.4 g CNP/kg of feed. Compared to the control, total bacterial load and coliform count decreased significantly (P&amp;lt; 0.05) by supplementing 0.4 g CNP in the diet. The dressing weight, breast weight and abdominal fat % were altered in birds receiving dietary 0.4 g CNP/kg. The treatment with CNP at 0.4 g/kg feed enhanced the broiler meat quality by increasing the values for water holding capacity (WHC), ABTS [2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)], DPPH (2,2-Diphenyl-1-picrylhydrazyl) while reducing the thiobarbituric acid reactive substances (TBARS) value. Based on the results above, it could be concluded that CNP supplementation at 0.4 g/kg is recommended as a beneficial feed additive for broiler chickens.

From the Gorgon's blood to Behring's Blutserumtherapie: A long path towards serum therapy.

At the end of the 19th century medicine was turned upside down by the development of serum therapy, a great therapeutic revolution as was vaccination a few years earlier. Many serums were developed, the most famous being the German doctor Emil von Behring's diphtheria serum, which saved countless children's lives from this dreadful disease. The discovery of the serum therapy principle, allowed by the progressive understanding of humoral immunity, occurred both in Germany and France, almost at the same time. Interestingly, this principle arose from two different intellectual paths, reviving the age-old opposition between mechanism and vitalism: while Behring came to this discovery reasoning as a chemist, French researchers Charles Richet and Jules Héricourt behaved as physiologists, focusing on the role of the host in the host-pathogen interaction. However, we should maybe consider that serum therapy history begins much earlier. Great forerunners must not be forgotten, especially researchers who investigated the field of immunity as soon as in the very beginning of the microbiological revolution; but also many people throughout centuries who tried to cure diseases with blood: as a transfer of blood serum, serum therapy also lies in the tradition of blood transfusion.

Sequence variants influencing the regulation of serum IgG subclass levels

Immunoglobulin G (IgG) is the main isotype of antibody in human blood. IgG consists of four subclasses (IgG1 to IgG4), encoded by separate constant region genes within the Ig heavy chain locus (IGH). Here, we report a genome-wide association study on blood IgG subclass levels. Across 4334 adults and 4571 individuals under 18 years, we discover ten new and identify four known variants at five loci influencing IgG subclass levels. These variants also affect the risk of asthma, autoimmune diseases, and blood traits. Seven variants map to the IGH locus, three to the Fcγ receptor (FCGR) locus, and two to the human leukocyte antigen (HLA) region, affecting the levels of all IgG subclasses. The most significant associations are observed between the G1m (f), G2m(n) and G3m(b*) allotypes, and IgG1, IgG2 and IgG3, respectively. Additionally, we describe selective associations with IgG4 at 16p11.2 (ITGAX) and 17q21.1 (IKZF3, ZPBP2, GSDMB, ORMDL3). Interestingly, the latter coincides with a highly pleiotropic signal where the allele associated with lower IgG4 levels protects against childhood asthma but predisposes to inflammatory bowel disease. Our results provide insight into the regulation of antibody-mediated immunity that can potentially be useful in the development of antibody based therapeutics.

An Exploratory Analysis of ChatGPT Compared to Human Performance With the Anesthesiology Oral Board Examination: Initial Insights and Implications.

Chat Generative Pre-Trained Transformer (ChatGPT) has been tested and has passed various high-level examinations. However, it has not been tested on an examination such as the American Board of Anesthesiology (ABA) Standardized Oral Examination (SOE). The SOE is designed to assess higher-level competencies, such as judgment, organization, adaptability to unexpected clinical changes, and presentation of information. Four anesthesiology fellows were examined on 2 sample ABA SOEs. Their answers were compared to those produced by the same questions asked to ChatGPT. All human and ChatGPT responses were transcribed, randomized by module, and then reproduced as complete examinations, using a commercially available software-based human voice replicator. Eight ABA applied examiners listened to and scored the topic and modules from 1 of the 4 versions of each of the 2 sample examinations. The ABA did not provide any support or collaboration with any authors. The anesthesiology fellow's answers were found to have a better median score than ChatGPT, for the module topics scores (P = .03). However, there was no significant difference in the median overall global module scores between the human and ChatGPT responses (P = .17). The examiners were able to identify the ChatGPT-generated answers for 23 of 24 modules (95.83%), with only 1 ChatGPT response perceived as from a human. In contrast, the examiners thought the human (fellow) responses were artificial intelligence (AI)-generated in 10 of 24 modules (41.67%). Examiner comments explained that ChatGPT generated relevant content, but were lengthy answers, which at times did not focus on the specific scenario priorities. There were no comments from the examiners regarding Chat GPT fact "hallucinations." ChatGPT generated SOE answers with comparable module ratings to anesthesiology fellows, as graded by 8 ABA oral board examiners. However, the ChatGPT answers were deemed subjectively inferior due to the length of responses and lack of focus. Future curation and training of an AI database, like ChatGPT, could produce answers more in line with ideal ABA SOE answers. This could lead to higher performance and an anesthesiology-specific trained AI useful for training and examination preparation.

Fibroblast stromal support model for predicting human papillomavirus-associated cancer drug responses.

Human papillomavirus-related cancers (HPV+ cancers) remain a significant public health concern, and specific clinical approaches are desperately needed. In translating drug response data from in vitro to in vivo, the fibroblasts of the adjacent stromal support network play a key role. Our study presents the utilization of a fibroblast 2D co-culture system to better predict translational drug assessments for HPV+ cancers. We also suggest that this co-culture system should be considered for other translational approaches. Predicting even a portion of treatment paradigms that may fail in vivo with a co-culture model will yield significant time, effort, resource, and cost efficiencies.

50 Awardee Talk: The role of non-traditional inheritance patterns among Bos indicus-Bos taurus crosses for global beef value chain considerations

Abstract Bos indicus-Bos taurus crosses have been effectively used in many production environments globally. Substantially higher amounts of hybrid vigor have been reported in these F1 crosses as compared with those involving only Bos taurus or only Bos indicus breeds. However, for many years, and in many environments, “unusual” aspects of calf growth have been documented among these crosses. Among F1 crosses, Bos indicus-sired calves (Bos indicus x Bos taurus) have birth weight 8 to 12 kg heavier than contemporary Bos taurus-sired calves (Bos taurus x Bos indicus); additionally, pronounced sexual dimorphism occurs where males may weigh 5 to 10 kg more than females from Bos indicus sires (Bos taurus dams) but 1 kg or less from Bos taurus sires (Bos indicus dams) in both natural service and embryo transfer calves. These reciprocal differences have been reported for fetal and placentome size mid gestation. Reciprocal differences in birth weight have also been reported among backcrosses and F2 calves, and also exist for longer gestation length from Bos indicus sires (Bos taurus dams). Increased birth weight and gestation length occur when more Bos indicus is present in the sire vs. the dam, and when Bos indicus influence is inherited through the sire vs. the dam. There is speculation that sex-specific genomic imprinting may be involved for reciprocal birth weight differences that are analogous to mouse interspecific crosses for litter weight. Reports of hybrid vigor retention among inter se matings after the F1 have been variable historically, and this variation may relate to how the foundational F1 parents were produced (Bos indicus- vs. Bos taurus-sired). Breed-of-origin haplotype on BTA5 has been shown to affect heifer pregnancy and rebreeding among F2 Nellore-Angus cows where Nellore-derived haplotypes had much lower performance compared with Angus-derived and hybrid haplotypes. Disproportionate calf sex has been reported among reciprocal F2 Nellore (Bos indicus)-Angus (Bos taurus) with 66.6% male calves from Nellore x Angus F1 sires. When retrospectively evaluating historical literature for reciprocal crosses, it appears that several instances occur with disproportionate calf sex percentage, and this phenomenon warrants further study. The extent that potential reciprocal differences may affect post-weaning growth and carcass traits has not been widely evaluated. Recent investigations have shown reciprocal F1 differences for some cell-mediated immune responses regarding T-lymphocyte proliferation, but no differences for humoral response. There needs to be additional research that completely characterizes the X chromosome of Bos indicus and Bos taurus origins and their roles for multiple production traits. There also needs to be more investigation of non-additive and parent-of-origin effects in genetic evaluations of Bos indicus-Bos taurus composites, particularly with desired first-generation breed percentage. Improved utilization of Bos indicus cattle and crosses will be instrumental in meeting global dietary protein demands.

Clinical Validation of the Nursing Diagnosis 'Inadequate Health Self-Efficacy' in People With Hypertension.

To test the clinical validity of clinical indicators and causal relationships of aetiological factors of the new nursing diagnosis of inadequate health self-efficacy in people with hypertension. The diagnosis of inadequate health self-efficacy has both theoretical and content validity. However, a clinical validation study is needed to establish an appropriate framework for distinguishing individuals who manifest this unique human response. The study adopts a cross-sectional clinical validation approach, adhering strictly to the STROBE guidelines throughout its design and implementation. Naturalistic sampling was used to identify 302 adults diagnosed with hypertension. Their data were subjected to latent class analysis, which facilitated the identification of a comprehensive set of clinical indicators that demonstrated better diagnostic accuracy and established posterior probabilities to guide the inference of inadequate health self-efficacy. In addition, logistic regression analysis was used to assess the magnitude of the impact of aetiological factors. The prevalence of inadequate health self-efficacy was 76.61%. Among the 13 indicators examined, seven demonstrated notable sensitivity: 'risk-prone health behaviour', 'failure to take action that prevents health problems', 'inadequate self-control', 'avoidance behaviours', 'negative health self-perception', 'inadequate health-related quality of life' and 'difficulty feeling good about adopting a healthy lifestyle'. Additionally, two indicators showed high specificity: 'difficulty feeling good about adopting a healthy lifestyle' and 'inadequate adherence to treatment regimen'. Notably, 15 aetiological factors were identified as significantly associated with an increased risk of inadequate health self-efficacy. A clinical framework consisting of eight clinical indicators and 15 aetiological factors was developed to characterise inadequate health self-efficacy in individuals with hypertension. Clinical validation provides insight into the precision of clinical indicators and the magnitude of the effect of putative causal elements, thereby facilitating identification and tailored intervention for individuals with hypertension and inadequate health self-efficacy.