Mitigating the underlying causes of differential immunity from rotavirus infection and vaccination requires a comprehensive understanding of rotavirus natural history and immune response. A US-based birth cohort provides a unique opportunity to longitudinally examine rotavirus immune response. Serum was collected from children at birth (cord blood), 6 weeks, and 6, 12, 18 and 24 months of age. Maternally-derived rotavirus-specific IgG (birth samples) and rotavirus-specific IgA and IgG (6-week through 24-month samples) were measured via enzyme immunoassay. Stool samples were collected weekly and during episodes of acute gastroenteritis to identify rotavirus infections. Serum samples prior to and after infection or vaccination were compared to assess their impact on IgA geometric mean titers (GMT). A Generalized Estimating Equation was fit to identify predictors of IgA GMT. The median change in IgA GMT from vaccination comparing 6-week and 6-month samples was 38.4 U/mL [IQR 4.4-138.1]. The change in IgA following infection varied widely (median=64.5 [IQR-2.0-192.9]) with pre- and post-infection GMTs of 35.7 U/mL (95% CI:21.4-59.6) and 102.6 U/mL (95% CI:41.1-256.0), respectively. In adjusted analyses, IgA GMT was higher when mother and/or child was a secretor (ratio of means (RoM)=1.3, 95% CI: 1.1-1.4) compared to non-secretor mother-child pairs. Being fully vaccinated was more strongly associated with IgA GMT in the first (RoM=1.9, 95% CI: 1.7-2.0) compared to the second year of life (RoM=1.3, 95% CI: 1.0-1.7). This longitudinal assessment of rotavirus-specific immune response contributes to our understanding of multifactorial drivers of rotavirus immunity in a post-vaccine setting.

Background. One of the key pathogenetic mechanisms in myelodysplastic syndrome (MDS) is a disruption of the hematopoietic stem cell microenvironment, which is accompanied by changes in the secretion of pro-inflammatory cytokines, in particular tumor necrosis factor alpha (TNF). Given the immunoinflammatory nature of MDS pathogenesis, the use of immunomodulatory drugs, in particular lenalidomide, has shown clinical efficacy in low-risk patients. Objective: to assess the clinical and hematological status and colony-forming activity of bone marrow cells in patients with MDS and refractory anemia with excess blasts 1 (RANB-1) depen­ding on the concentration of TNF in the blood serum. Materials and methods. Twenty-seven patients receiving lenalidomide were examined. Serum TNF level was determined with enzyme-linked immunosorbent assay using standard production kits. The analysis was performed on an enzyme immunoassay analyzer Multiskan EX ( = 430 nm). Plasma from healthy donors served as a control. ­Results. It was found that a decrease in TNF is accompanied by clinical improvement, increased erythropoiesis and increased co­lony formation in vitro. Serum TNF concentration significantly decreases in patients with MDS and RANB-1 when achieving a complete or partial response to lenalidomide therapy. TNF level demonstrates high predictive accuracy (AUC = 1.00) for differentiating response to treatment. Functional activity of progenitor cells (CFU-GM) and the level of CD34+/CD117+ in the bone marrow are inversely related to TNF. It is noteworthy that in the group of patients with MDS and RANB-1 who did not respond to lenalidomide therapy, a deterioration in the general condition was noted due to worsening anemia, which can be assessed as a clinical situation for correction of the therapeutic route in order to prevent emergencies. Conclusions. The obtained results confirm the feasibility of including TNF and colony formation in the set of diagnostic markers for risk stratification and predicting the course of MDS.

To evaluate the diagnostic sensitivity of real-time PCR to detect Coccidioides spp DNA from oropharyngeal, nasal cavity, and conjunctival swab samples in dogs with pulmonary coccidioidomycosis. This was a prospective cohort study of client-owned dogs with newly diagnosed pulmonary coccidioidomycosis from January 2024 to October 2024. Sterile cotton swabs were gently rubbed within the nasal cavity and against the mucosa of the oropharynx and conjunctiva during the initial investigation for respiratory tract signs. Swabs were tested for Coccidioides spp, Blastomyces spp, Histoplasma spp, Pneumocystis jirovecii, and Pneumocystis canis DNA by multiplex quantitative PCR. Antibody serology assays (ie, agar gel immunodiffusion and enzyme immunoassay) were performed at a single reference diagnostic laboratory. 24 dogs were included. Seventy-two PCR tests (nasal cavity, n = 24; oropharyngeal, 24; and conjunctival, 24) were performed, and none of the tests positively detected Coccidioides spp DNA. Forty-six percent (11 of 24) and 71% (17 of 24) of dogs had positive detection of anti-Coccidioides spp immunoglobulin M and G antibodies with agar gel immunodiffusion, respectively. Seventy-nine percent (19 of 24) of dogs had positive detection of immunoglobulin G antibodies with enzyme immunoassay. Respiratory tract signs improved (50%; n = 12) or completely resolved (42%; 10) after approximately 3 months of PO fluconazole administration. Coccidioides spp DNA was not detected in any swab sample from the upper respiratory tract. Quantitative PCR to detect Coccidioides spp DNA from the nasal cavity, oropharynx, and conjunctiva using sterile cotton swabs provides no added diagnostic value in dogs with pulmonary coccidioidomycosis.

Our goal was to investigate immunoglobulin G1 (IgG1) and immunoglobulin G3 (IgG3) antibody responses to Chlamydia trachomatis proteins Pgp3 and Hsp60 in males of subfertile couples and to explore the association of these antibodies with semen parameters and male factor infertility. Serum samples were collected from 256 male partners of subfertile couples. Serum IgG1 and IgG3 antibodies to C trachomatis Pgp3 and Hsp60 were measured using enzyme immunoassays. Semen samples were analyzed for volume, sperm concentration, and motility according to World Health Organization criteria. Altogether, 74 (29.8%) men were seropositive to either C trachomatis Pgp3 IgG1 or IgG3, and 67 (27.0%) to either Hsp60 IgG1 or IgG3. Chlamydia trachomatis Pgp3 IgG1 and IgG3 antibodies were associated with impaired sperm motility (asthenozoospermia) (18.6% vs 6.3%, P = .006 for Pgp3 IgG1; and 21.4% vs 8.0%, P = .03 for Pgp3 IgG3). After adjusting for smoking, alcohol risk consumption, and body mass index, the association between serum C trachomatis Pgp3 IgG1 seropositivity and asthenozoospermia remained statistically significant (odds ratio, 3.0 [95% confidence interval, 1.12-8.01]; P = .03). The presence of Hsp60 IgG1 antibody was associated with a higher teratozoospermia index (1.47 ± 0.15 vs 1.39 ± 0.16; P = .001). Our results suggest that prior Chlamydia trachomatis infection, as indicated by Pgp3 seropositivity, may negatively impact male fertility potential by affecting sperm motility and morphology.

Cortisol, a stress biomarker, may be associated with lower physical activity (PA) tolerance. This study examined the associations between stress (both subjective and objective) and PA in patients with heart failure (HF) with systolic, diastolic, and combined (systolic + diastolic) HF phenotypes. In this cross-sectional study, patients with a diagnosis of heart failure/cardiomyopathyreported stress levels subjectively using the Perceived Stress Scale (PSS-10). Hair strands, collected from the posterior vertex of each participant's head, were processed using enzyme immunoassay to measure hair cortisol concentration (HCC; pg/mg). PA was assessed using the International Physical Activity Questionnaire Short Form (IPAQ-SF). Participants included 28 systolic, 7 diastolic, and 11 combined HF patients. PA levels were low across all groups, with combined HF patients showing a trend toward the lowest PA levels. Combined HF patients also demonstrated a higher trend toward perceived stress (16.9+5.1) and HCC (15.3+17.9 pg/mg). HCC did not show a strong associated with perceived stress in any group. However, HCC was strongly and positively associated with PA among patients with diastolic HF (r = 0.91), but not among patients with systolic HF (r = -0.09) or combined HF (r = 0.26). A potential dissociation between psychological and physiological stress was noted. PA was significantly and positively associated with HCC in diastolic HF, suggesting that PA may serve as a physiological stressor. Both subjective and objective markers of stress were higher in patients with combined HF. More research is needed to investigate these findings.

Correction for Suzuki et al., "Evaluation of fully automated chemiluminescent enzyme immunoassays for hepatitis B core-related antigen components, phosphorylated and non-phosphorylated hepatitis B core antigens: clinical significance and dynamics during hepatitis B e antigen seroconversion".

Cupressales species are widely planted in urban landscapes, where their pollen represents a growing source of biological air pollution and contributes to seasonal allergies. Linking their flowering dynamics with atmospheric allergen loads is essential for understanding ecological flows of airborne particles in cities. We aimed to quantify interspecific differences in allergenic potential among common urban Cupressales taxa and to determine how flowering periods and pollen production shape airborne allergen levels. We specifically assessed variation in Cup a 1 homolog abundance to evaluate its relevance for landscape-scale allergy risk. The study was conducted in Poznań (Poland) during the 2023-2024 pollen seasons. Phenological observations, aerobiological monitoring, and immunoenzymatic assays were combined to relate flowering timing, atmospheric pollen concentrations, and allergen content across ten Cupressales species. Species differed substantially in flowering phenology, pollen output, and allergen levels. Taxus baccata, Thuja occidentalis, and Th. plicata released large amounts of pollen but contained low levels of Cup a 1 homologs. Conversely, Juniperus taxa and Callitropsis nootkatensis showed consistently high allergenicity, contributing disproportionately to peak atmospheric allergen loads despite moderate pollen release. These patterns indicate that allergen exposure risk is driven not only by pollen abundance but also by species-specific allergen potency. Urban planting dominated by high-allergenicity taxa may therefore intensify allergy symptoms at the neighbourhood scale. Our findings demonstrate that allergenicity metrics can inform urban vegetation planning. Selecting species with favourable phenological and allergenic profiles may help reduce airborne allergens and support healthier, more resilient urban landscapes.

Abstract Background: Clinical assessment of breast cancer progression and therapeutic response typically relies on qualitative immunohistochemistry results for ER, PR, and HER2 proteins, which may be improved by adherence to ASCO/CAP Guidelines. To identify more robust, clinically relevant molecular markers, we conducted in silico analyses of a comprehensive, de-identified database that integrates quantified protein biomarker levels, qPCR, and microarray gene expression data, as well as long-term clinical outcomes. Because TNBC represents an aggressive disease that is difficult to treat, our multifarious investigation focuses on differentiating biomarkers for this subtype that improve clinical outcomes. Methods: Biomarker Triads were created from ER and PR proteins quantified by radio-ligand binding (n = 13,966) and enzyme immunoassays (n = 4,408), expressed as fmol/mg cytosol protein. HER2 protein was quantified in 504 and 901 specimens using either ELISA or EIA, respectively, and expressed as HNU/mg membrane protein. Clinical data were available for 1,194 ER/PR cases, and complete ER/PR/HER2 protein Triads were available for 189 of these. However, survival outcomes (PFS and OS) were only available for a subset of these patients. Expression levels of these genes and 82 other cancer-associated genes were validated by RTqPCR of 580 specimens after RNA extraction and purification. Microarray analysis (∼22,000 genes) was conducted on RNA from each of 247 LCM-isolated carcinoma samples. Biomarker Triads were defined as ER+/PR+/HER2+ (TPBC) or ER-/PR-/HER2- (TNBC), using either protein levels or cognate gene expression (ESR1, PGR, ERBB2). Differential gene expression was assessed using nonparametric tests and Spearman correlations in R (version 4.4.2). For the subset of patients with survival data, exploratory Kaplan-Meier and Cox proportional hazards models were used to assess associations between gene expression and progression-free survival (PFS) or overall survival (OS). Results: Neither ER/PR assay platform influenced ER+/PR+ or ER-/PR- status, and neither ER nor PR levels were associated with HER2 expression. TPBC biopsies showed elevated expression of BL2 (CADM1), CAXII, ERBB2, and ERBB4 compared to TNBC, supporting their use in subtype definition. TPBC also exhibited increased IL10 and VEGFATV2, while TGFB1 was downregulated in both TPBC and ER+ cases. BRCA1/2 expression was reduced in ER-/PR- tumors, while NAT1/2 and COMT were elevated in ER+, ER+/PR+, and TPBC samples. FOXA1 and ART3 emerged as potential targets from gene-based Triad definitions using microarray data. In a subset of cases with available clinical outcomes, exploratory survival analysis suggested that TPBC triads were associated with more favorable PFS and OS compared to TNBC, and elevated expression of certain genes (e.g., ESR1, FOXA1) trended with improved survival outcomes. Conclusions: This integrative dataset, which links rigorously collected clinical biopsies, precise biomarker protein quantification, and gene expression profiles from LCM-procured carcinoma cells, enabled the discovery of genes differentially expressed across biomarker-defined subtypes. Biomarker Triads based on protein and gene expression levels in primary breast tissue biopsies revealed a landscape of genomic diversity in carcinomas, which improved molecular subtype classification. Select genes, including the transcription factor FOXA1 and ART3, reported to be involved in cell migration, represent probable targets for drug development and/or companion diagnostics in ER+ and triple-negative breast carcinomas. Citation Format: M. W. Daniels, J. L. Wittliff. Genomic Diversity of Breast Carcinomas Exhibiting Biomarker Triads Derived from Quantified Levels of ER, PR and HER2 Proteins or from Expression of Their Cognate Genes [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2025; 2025 Dec 9-12; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2026;32(4 Suppl):Abstract nr PS4-05-20.

Abstract The ability to monitor response to therapy and disease progression in patients with metastatic breast cancer (MBC) is a major step in patient management. Sequential imaging remains the method of choice for the assessment of disease status and monitoring of disease response or progression. Serum levels of tumor-associated biomarkers such as CA15-3 and CEA are also measured to follow MBC patients’ disease status. However, it may also be valuable to measure levels of novel biomarkers that have been shown to be overexpressed in breast cancer tumors and play a role in drug resistance and for which therapeutic development is on-going. Progranulin, also called Glycoprotein 88kDa (PGRN/GP88) is an autocrine growth factor overexpressed in several cancers including breast cancer. PGRN plays a significant role in breast tumorigenesis. PGRN/GP88 overexpression in invasive ductal carcinoma (IDC) is associated with malignant phenotype, estrogen (ER) independence, increased proliferation, survival, angiogenesis, and drug resistance. High PGRN/GP88 tumor expression measured by immunohistochemistry in ER receptor positive IDC is an independent prognostic marker associated with increased risk of recurrence and mortality. Clinical studies have demonstrated that serum PGRN levels determined by sandwich enzyme immunoassay are elevated in breast cancer (BC) patients, compared to healthy individuals. In MBC patients, low serum PGRN levels correlate with increased overall survival. Based on these observations, an IRB approved, prospective study was established at the University of Maryland Greenebaum Comprehensive Cancer Center to examine serum PGRN/GP88 levels in association with disease status as measured by RECIST 1.1 criteria in MBC patients receiving standard of care therapies. A total of 103 MBC patients with measurable or evaluable metastatic disease will be consented and enrolled. For inclusion, patients must have been re-staged within 4 weeks of study entry and continue or begin new anticancer therapy. Currently, 50 patients have been enrolled with blood samples collected every 2-3 months or whenever there is a disease event such as clinical or imaging progression. Standard of care (SOC) laboratory assessments and radiographic imaging/staging will be done on study to complement blood collection to measure PGRN/GP88. The samples are stored at -70C until evaluation of PGRN/GP88 using an enzyme linked immunoassay developed by A&G Pharmaceutical. The serum PGRN/GP88 levels will then be correlated with disease response by RECIST 1.1 (progression, stable for < 6 vs >=6 months, partial/complete response), progression free survival (PFS), duration of response (DOR) and/or need for change of therapy. In this study the decision about the change of cancer therapy will be based on the SOC approaches and will not be guided by PGRN/GP88 results. This study is supported by grant R44CA210817 from the National Cancer Institute to Ginette Serrero and the University of Maryland Greenebaum Comprehensive Greenebaum Cancer Center. Citation Format: K. H. Tkaczuk, K. Hu, P. Y. Rosenblatt, N. Tait, Y. BinBin, G. Serrero. Prospective longitudinal study of circulating serum progranulin (PGRN/GP88) levels and its association with tumor response to therapies in patients with metastatic breast cancer (MBC) [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2025; 2025 Dec 9-12; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2026;32(4 Suppl):Abstract nr PS5-09-03.

Lipemia is a common comorbidity in horses with obesity or equine metabolic syndrome, but the impact of lipemia on the measurement of insulin and adiponectin has not been evaluated. To evaluate endogenous and exogenous lipemic interference with equine insulin and adiponectin measurements via several commercial assays. Endogenous lipemia was evaluated using plasma and serum samples with triglyceride concentrations of < 40, 40-250, 250-500, 500-1000, and > 1000 mg/dL (n = 6 each). Sample insulin concentrations were determined via fluorescence enzyme immunoassay (FEIA), ELISA, and lateral flow assay (LFA). Test agreement was assessed using Pearson's correlation coefficient, Passing-Bablok regression, and Bland-Altman analysis. Exogenous lipemia was evaluated using pools of serum, plasma, and whole blood (n = 5 each) spiked with Intralipid 20% to triglyceride concentrations of 0, 50, 100, 250, 500, and 1000 mg/dL. Insulin concentrations were measured via FEIA, ELISA, and LFA, and adiponectin concentrations via immunoturbidometric assay (ITA). Interferograms were created and a Kruskal-Wallis test was used to compare bias between triglyceride concentrations. With endogenous lipemia, agreement between the three assays was excellent (r > 0.90) with no appreciable impact of triglyceride concentration. For exogenous lipemia, significant (p < 0.05) negative interference was observed at triglyceride concentrations of 1000 mg/dL with the insulin ELISA using plasma. No significant interference was found for the insulin ELISA or FEIA using serum, insulin LFA using whole blood or plasma, or adiponectin ITA using serum. Falsely low insulin values may be obtained at high triglyceride concentrations with the insulin ELISA using plasma.

Constrained bicyclic peptides (Bicycle molecules) with high affinity for biological targets have emerged as potentially powerful therapeutic agents, particularly for the in vivo targeting of cancer receptors. However, their antibody-mimetic properties have yet to be explored for use in diagnostic immunoassays. These synthetically derived compounds serve as biorecognition scaffolds that allow for facile site-selective modification and large-scale production. A phage display screen against various constructs of the SARS-CoV-2 nucleocapsid (N) protein identified several Bicycle molecules with binding affinities ranging from the micromolar to the low nanomolar range. These Bicycle molecules were validated in the development of enzyme- and nanozyme-linked immunosorbent assays, as well as enzymatic and colorimetric nanoparticle-based lateral flow immunoassays (LFIA) for the detection of ultralow concentrations of the SARS-CoV-2 N protein. We envision that these moieties enable robust, cost-effective, and large-scale development of ultrasensitive biosensors for a diverse range of biomarkers by leveraging their high binding affinity, minimalistic scaffold, and synthetic accessibility.

Background: Rotaviruses are among the most common causal pathogens of severe dehydrating diarrhoea in children. Little is known about the burden of rotavirus diarrhoea in Gabon. Aim: This study aimed to determine the proportion of rotavirus infection in children under 5 years with diarrhoea in Lambaréné and seen at the hospital and factors associated with rotavirus infection. Setting: The data used in this study were collected between February 2020 and February 2021 in children presenting with acute diarrhoea in the Albert Schweitzer Hospital paediatric ward. Methods: A cross-sectional study was carried out. Stool samples were tested for rotavirus antigens using the rotavirus Standard Diagnostic (SD) BIOLINE Rota and Adeno enzyme immunoassay detection kit. Results: A total of 178 children were included in the study. The proportion of rotavirus infection was 22% (n = 39/178; 95% confidence interval [CI]: 16% – 29%). In the multivariate analysis, the rotavirus was independently associated with dehydration (adjusted odds ratio [aOR] = 2.65; 95% CI: 1.09–6.86), vomiting (aOR = 3.15; 95% CI: 1.29–8.25), lethargy (aOR = 3.12; 95% CI: 1.16–8.71) and hospitalisation (aOR = 4.63; 95% CI: 1.7–13.65). Conclusion: Rotavirus infection was associated with severe diarrhoea and hospitalisation. This study shows the need to integrate and support free rotavirus vaccination into the expanded vaccination programme in Gabon. Contribution: This study provides evidence that could guide public health strategies and inform vaccine policies that could ultimately reduce the burden of rotavirus-associated diarrhoea in children.

Accurate molecular epidemiology relies on recovering Clostridioides difficile from clinical specimens and is essential for informing public health responses, characterizing strains that cause disease, and tracking strain prevalence. We performed a cross-sectional study of the recovery of C. difficile isolates from test-positive stools received January 2020 through December 2022 from the Colorado and Georgia Emerging Infections Program sites. Multivariable logistic regression models determined the adjusted odds ratio (aOR) and 95% confidence intervals (CIs) for factors which may influence C. difficile recovery in specimens that tested positive by either a dedicated or multiplex polymerase chain reaction (PCR) as part of either a reverse testing algorithm (PCR-positive, arbitrated by toxin enzyme immunoassay [EIA]) or a PCR-only testing protocol. Whole-genome sequencing was performed to determine the multilocus sequence types. C. difficile was recovered by culture from 84.3% (1,527/1,811) of all specimens. In specimens tested with a reverse testing algorithm, C. difficile culture recovery was less likely with multiplex versus dedicated PCR (aOR: 0.53; 95% CI: 0.34-0.85) and more likely in toxin EIA-positive versus toxin EIA-negative specimens (aOR: 8.07; 95% CI: 4.10-18.31). Recovery of C. difficile from stool cultures in PCR-only protocol cases was less likely when C. difficile was detected by multiplex PCR (aOR: 0.16; 95% CI: 0.03-0.55). ST1 was associated with toxin EIA-positive specimens, whereas ST2/110 was associated with toxin EIA-negative specimens. These findings can be used to maximize C. difficile surveillance programs, thereby enhancing the ability to generate robust molecular epidemiology data to inform prevention and control efforts.IMPORTANCEPublic health surveillance of Clostridioides difficile is essential for tracking strains, understanding transmission, and informing public health strategies. The foundation of the surveillance is the successful recovery of C. difficile from clinical specimens for molecular typing. This study reveals that the choice of diagnostic test significantly impacts the ability to recover C. difficile by culture. These data reveal that culture recovery was lower from specimens that tested positive by syndromic multiplex polymerase chain reaction (PCR) panels compared to dedicated C. difficile PCR assays. Furthermore, recovery was more than eight times more likely from toxin enzyme immunoassay (EIA)-positive specimens than from toxin EIA-negative specimens, with high-virulence strains, such as PCR-ribotype 027, being associated with toxin EIA-positive results. These findings demonstrate that common diagnostic practices could introduce biases into surveillance data, potentially misrepresenting the true prevalence and epidemiology of clinically important strains. Understanding these factors is crucial for optimizing surveillance programs to generate accurate molecular epidemiologic data.

Environmental changes, such as transfers and social introductions, can be major sources of stress in wild animals. Monitoring the physiological response through glucocorticoids is an effective tool to assess how animals cope with and adjust to these changes. In this study, we followed the habituation and social integration of Suzie, a former pet and entertainment chimpanzee confiscated and transferred to a primate rescue center. Our objectives were to (1) monitor her fecal glucocorticoid metabolite (FGM) levels throughout rehabilitation, and (2) investigate how association-related factors (e.g. number of individuals, ages of association partners) influenced her FGM levels and social behaviors during associations with the resident chimpanzees. A total of 169 fecal samples from Suzie were collected over 10months and quantified using enzyme immunoassay, along with 38 opportunistic samples from the resident chimpanzees. During association sessions, data on aggressive and affiliative events were recorded. Suzie's FGM levels ranged from 11.01 to 113.04ng/g of feces. After a four-month adjustment period (118days) before returning to basal hormone activity, her FGM concentrations aligned with individual mean levels of the resident chimpanzees (range: 24.19 ± 7.99 to 29.69 ± 8.93ng/g of feces). The presence of a broker individual during associations impacted the likelihood of affiliative events (p < 0.01). Furthermore, early social housing conditions of the resident chimpanzees influenced social dynamics, with affiliative behaviors being more likely to occur while interacting with individuals who were housed in social groups during infancy, and aggressive events being more likely to occur while interacting with those housed alone (p < 0.05). Our findings underscore the value of non-invasive hormone monitoring and behavioral assessments to better understand individual experiences.

Diagnosing atypical IgG4-mediated anti-glomerular basement membrane (anti-GBM) disease is challenging because conventional serological assays poorly detect IgG4 antibodies. Here we study which commercial assays are affected and we describe proof-of-concept of improved IgG4anti-GBM detection. To investigate the scope of the diagnostic dilemma detecting IgG4anti-GBM antibodies, serum from a patient with atypical IgG4-mediated anti-GBM was distributed to 36 laboratories participating in the Dutch External Quality Assessment (EQA) program. To improve IgG4anti-GBM detection in the fluorescent enzyme immunoassay (FEIA), the standard anti-IgG conjugate was replaced with anti-IgG4 conjugate. We report the diagnostic delay of a patient with atypical anti-GBM disease who presented with an indolent disease course. Histopathology comprised hallmarks of classic anti-GBM disease including bright linear IgG deposits along the GBM but without the typical findings of diffuse crescentic and necrotizing glomerulonephritis. IgG anti-GBM test results were repeatedly negative. Histopathological subclass analysis demonstrated that the linear IgG deposits were predominantly IgG4. All 36 Dutch EQA-participants reported negative anti-GBM test results, demonstrating that the diagnostic challenge of detecting IgG4anti-GBM is broadly applicable across multiple diagnostic assays. A minor modification to the manufacturer's standard protocol of the FEIA anti-GBM dramatically improved the assay's performance for measuring antibodies of the IgG4 isotype. Using the modified IgG4-GBM test, we were able to diagnose and monitor one more patient with atypical IgG4-mediated anti-GBM disease. Here we demonstrate proof-of-concept of a modified IgG4anti-GBM blood test allowing serological confirmation of atypical anti-GBM disease and sensitive monitoring of therapy response.

Comparative evaluation of SARS-CoV-2 antigens as capture and detection elements in an in-house antigen-based ELISA for COVID-19 total antibody detection.

Differential profiles of GH, IGF-1, and fructosamine in follicular fluid and plasma of cyclic mares.

Clostridioides difficile infection (CDI) poses a diagnostic challenge in patients with inflammatory bowel disease (IBD), as symptoms and endoscopic findings frequently overlap with those of disease flares, and asymptomatic colonization is relatively common. Consequently, positive microbiological test results do not always indicate active infection. We report the case of a 53-year-old woman with ulcerative colitis (UC) who presented with worsening bloody diarrhea and abdominal pain during maintenance therapy with infliximab. At initial admission, she had clinically significant diarrhea with elevated inflammatory markers. Stool testing demonstrated concordant positivity for glutamate dehydrogenase (GDH) antigen and toxin A/B enzyme immunoassay, supporting a diagnosis of active CDI concomitant with a UC flare. Treatment with oral vancomycin led to clinical improvement. Three weeks later, the patient re-presented with mild recurrent diarrhea. Repeat stool testing showed discordant results, including negative toxin A/B but positive GDH, polymerase chain reaction, and culture. Given the mild symptoms, normal inflammatory markers, and spontaneous symptom resolution without further antimicrobial therapy, these findings were interpreted as colonization or residual test positivity rather than recurrent CDI. This case illustrates key principles in CDI diagnosis: testing should be guided by the presence of clinically significant diarrhea, multistep diagnostic algorithms must be interpreted in the clinical context, and positive molecular or culture-based results in IBD patients do not necessarily reflect active infection. Symptom-based assessment is essential to avoid overdiagnosis and unnecessary antimicrobial treatment in patients with UC and suspected CDI.

Equine oviduct-specific glycoprotein is modulated by hormones and sperm cells.

The aim of the work was to determine the effect of feeding a liposomal vitamin complex with zinc gluconate under heat stress (HS) on the concentration of individual hormones and the quality of boar sperm. The experiment was conducted on nine clinically healthy breeding boars, aged 2–4 years of Landrace, Pietren and Duroc breeds. Three stages of the study were conducted, each lasting 30 days, in which the selection of material and its analysis were similar: 1) under normal thermal conditions (&lt;23 °C); 2) under HS conditions (25–30 °C); 3) under feeding a complex liposomal supplement under HS conditions (25–30 °C). In the third stage of the research on the background of HS, all boars were individually given a feed additive in the form of a liposomal emulsion for 30 days, which included vitamins A, D3, E, and C with zinc gluconate in a dose of 2 ml. At the end of each stage of the experiment, blood samples were taken from the experimental boars. The concentration of testosterone, cortisol, and thyroxine in blood plasma was determined by the enzyme-linked immunoenzymatic assay method. After completing each stage, ejaculates were taken from the boars by manual method twice a week for two weeks. The parameters of motility and morphology of germ cells, the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were determined. It was found that under the influence of HS in the blood plasma of boars, the level of cortisol (P&lt;0.01) and thyroxine (P&lt;0.05) increased, while the concentration of testosterone significantly decreased (P&lt;0.05). The negative effect of HS on spermatogenesis is confirmed by a significant (P&lt;0.001) decrease in the concentration of testosterone in the plasma of sperm. Moderate HS reduces the overall motility of sperm of breeding boars (P&lt;0.001) and the activity of germ cells with rectilinear translational movement (progressive motility; P&lt;0.01), and doubles the percentage of degenerated sperm (P&lt;0.001). Under the influence of HS, a decrease in the activity of GPx and CAT (P&lt;0.001) is observed in the sperm of boars against the background of a slight increase in the activity of SOD. After feeding the liposomal supplement under the action of HC, the concentration of cortisol and thyroxine in the blood of boars significantly (P&lt;0.05–0.01) decreased, the level of testosterone in the blood and semen of boars significantly increased (P&lt;0.05–0.001). This led to an improvement in sperm motility and their morphological characteristics: the total sperm motility significantly increased (P&lt;0.01) and the activity of sperm with progressive motility (p&lt;0.01) with a simultaneous decrease in the percentage of degenerated sperm (P&lt;0.01). At the same time, the activity of SOD significantly decreased (p&lt;0.01) with a simultaneous increase in the activity of GPx (P&lt;0.001) and CAT (P&lt;0.001).