Positive Serum Research Articles

Targeted serum proteome profiling reveals nicotinamide adenine dinucleotide phosphate (NADPH)-related biomarkers to discriminate linear IgA bullous disorder from dermatitis herpetiformis

Linear IgA bullous dermatosis (LABD) and dermatitis herpetiformis (DH) represent the major subtypes of IgA mediated autoimmune bullous disorders. We sought to understand the disease etiology by using serum proteomics. We assessed 92 organ damage biomarkers in LABD, DH, and healthy controls using the Olink high-throughput proteomics. The positive proteomic serum biomarkers were used to correlate with clinical features and HLA type. Targeted proteomic analysis of IgA deposition bullous disorders vs. controls showed elevated biomarkers. Further clustering and enrichment analyses identified distinct clusters between LABD and DH, highlighting the involvement of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Comparative analysis revealed biomarkers with distinction between LABD and DH and validated in the skin lesion. Finally, qualitative correlation analysis with DEPs suggested six biomarkers (NBN, NCF2, CAPG, FES, BID, and PXN) have better prognosis in DH patients. These findings provide potential biomarkers to differentiate the disease subtype of IgA deposition bullous disease.

Evaluation of the correlation of serological and intradermal allergen testing with clinical history in 29 dogs with atopic dermatitis.

Limited information exists about the correlation between clinical history and positive serum (SAT) and intradermal allergen test (IDAT) results in atopic dogs. To evaluate the correlation between clinical history and SAT/IDAT results in atopic dogs. Twenty-nine client-owned dogs with nonseasonal atopic dermatitis with or without seasonal exacerbation were enrolled. IDAT, SAT (immunoglobulin (Ig)M antibody capture enzyme-linked immunosorbent assay [MacELISA] with bromelain CCD inhibitor) and clinical information collected in a questionnaire regarding seasonal variations in pruritus affecting the dogs were performed on the same day. Two independent investigators (Inv A and Inv B) recorded IDAT results. The kappa coefficients agreement for positive IDAT scores between Inv A and B was substantial. The agreement between IDAT and SAT results at different ELISA absorbance units (EAU) cut-offs (>79 and ≥300) was slight and fair for both investigators, respectively. A higher agreement was observed between IDAT and SAT (≥300 EAU) than between IDAT and SAT (>79 EAU) with the exception of mite and flea allergens. There was a statistically significant association between clinical history and positive IDAT results for seasonal allergens (Inv A and Inv B, p = 0.016). There was no significance between positive SAT results and clinical history. Five (IDAT) and 12 of 13 (SAT) atopic dogs without clinical seasonal exacerbation showed positive results for seasonal allergens. The agreement between IDAT and SAT ≥300 EAU results was fair and the agreement between IDAT and SAT >79 EAU results was slight for all allergens. Only positive IDAT results significantly correlated with clinical history.

An explainable machine learning-based model to predict intensive care unit admission among patients with community-acquired pneumonia and connective tissue disease

BackgroundThere is no individualized prediction model for intensive care unit (ICU) admission on patients with community-acquired pneumonia (CAP) and connective tissue disease (CTD) so far. In this study, we aimed to establish a machine learning-based model for predicting the need for ICU admission among those patients.MethodsThis was a retrospective study on patients admitted into a University Hospital in China between November 2008 and November 2021. Patients were included if they were diagnosed with CAP and CTD during admission and hospitalization. Data related to demographics, CTD types, comorbidities, vital signs and laboratory results during the first 24 h of hospitalization were collected. The baseline variables were screened to identify potential predictors via three methods, including univariate analysis, least absolute shrinkage and selection operator (Lasso) regression and Boruta algorithm. Nine supervised machine learning algorithms were used to build prediction models. We evaluated the performances of differentiation, calibration, and clinical utility of all models to determine the optimal model. The Shapley Additive Explanations (SHAP) and Local Interpretable Model-Agnostic Explanations (LIME) techniques were performed to interpret the optimal model.ResultsThe included patients were randomly divided into the training set (1070 patients) and the testing set (459 patients) at a ratio of 70:30. The intersection results of three feature selection approaches yielded 16 predictors. The eXtreme gradient boosting (XGBoost) model achieved the highest area under the receiver operating characteristic curve (AUC) (0.941) and accuracy (0.913) among various models. The calibration curve and decision curve analysis (DCA) both suggested that the XGBoost model outperformed other models. The SHAP summary plots illustrated the top 6 features with the greatest importance, including higher N-terminal pro-B-type natriuretic peptide (NT-proBNP) and C-reactive protein (CRP), lower level of CD4 + T cell, lymphocyte and serum sodium, and positive serum (1,3)-β-D-glucan test (G test).ConclusionWe successfully developed, evaluated and explained a machine learning-based model for predicting ICU admission in patients with CAP and CTD. The XGBoost model could be clinical referenced after external validation and improvement.

Identification of a New B-Cell Epitope on the Capsid Protein of Avian Leukosis Virus and Its Application.

Avian leukosis virus (ALV) is an avian oncogenic retrovirus that can impair immunological function, stunt growth and decrease egg production in avian flocks. The capsid protein (P27) is an attractive candidate for ALV diagnostics. In the present study, a new hybridoma cell (1F8) stably secreting an anti-P27 monoclonal antibody (mAb) was developed. The mAb exhibited a high affinity constant (Ka) of 8.65 × 106.0 L/mol, and it could be used for the detection of ALV-A/B/J/K strains. Moreover, a total of eight truncated recombinant proteins and five synthetic polypeptides were utilized for the identification of the B-cell epitopes present on P27. The results revealed that 218IIKYVLDRQK227 was the minimal epitope recognized by 1F8, which had never been reported before. Additionally, the epitopes could strongly react with different ALV subgroup's specific positive serum and had a complete homology among all the ALV subgroups strains. Finally, a new sandwich ELISA method was created for the detection of ALV antigens, demonstrating increased sensitivity compared to a commercially available ELISA kit. These results offer essential knowledge for further characterizing the antigenic composition of ALV P27 and will facilitate the development of diagnostic reagents for ALV.

Seroprevalence and Endemic Status of Babesia ovis by Enzyme-Linked Immunosorbent Assay in East Azerbaijan Province, North-West of Iran.

Babesia ovis, an intraerythrocytic parasite carried by ticks and one of the most common subclinical ovine illnesses, was studied to ascertain its seroprevalence and endemic status in ram and ewe populations in East Azerbaijan Province, Iran, in lambs, yearlings, and adults of over two years of age. A total of 960 sheep from 10 cities were selected from Jan 2018 to Nov 2019. Blood samples were collected from each animal and tested for the presence of B. ovis antibodies by applying a developed enzyme-linked immunosorbent assay (ELISA) technique. Checkerboard titrations were used to determine the optimal dilution of the antigen using negative and positive control sera. To determine whether the disease is endemically stable, inoculation rates for each age group were also calculated. Correlation coefficients were calculated between age and infection rates and also between age and inoculation rates. The results revealed an average infection rate of 49.4% in East Azerbaijan Province. There was a positive correlation between the age of animals and susceptibility to infection except for lambs and yearlings, whereas there was no meaningful difference in exposure to B. ovis between rams and ewes. The negative correlation between age and inoculation rates indicates increased disease instability with age. Inoculation rate results revealed the endemically instable status of B. ovis in the studied area. High prevalence rates and endemically instable status of the disease suggest demand for vaccine development and implementation of appropriate control measures for ovine babesiosis to mitigate the associated economic losses.

A case of variant of GBS with positive serum ganglioside GD3 IgG antibody

BackgroundAcute bulbar palsy-plus (ABPp) syndrome is an unusual variant of Guillain-Barré syndrome (GBS). Anti-GT1a and anti-GQ1b antibodies have been reported in patients with ABPp, but without reports related to GD3 antibodies.MethodsClinical data of a patient diagnosed as ABPp syndrome were reviewed clinically. And we summarized the GBS patients with ABP and facial paralysis reported in the literature.ResultsWe reported a 13-year-old girl presented with asymmetric bifacial weakness, bulbar palsy and transient limb numbness, and had positive serum IgG anti-GD3 antibody. Through reviewing the GBS patients with ABP and facial paralysis reported previously, we found that facial palsy could be unilateral or bilateral. The bilateral facial palsy could present successively or simultaneously, and could be symmetrical or asymmetrical. Other common symptoms included ophthalmoplegia, sensory abnormality and ataxia. IgG anti-GT1a and IgG anti-GQ1b antibodies were the most frequent. Most of the patients had full recovery within two weeks to one year of follow-up.ConclusionsWe reported a patient with asymmetric bifacial palsy and bulbar palsy, which seemed to fit the diagnosis of ABPp syndrome. This was the first report of ABPp variant of GBS with positive serum ganglioside GD3 IgG antibody.

Sero-epidemiological study of brucellosis in cattle under pastoral/agro-pastoral and mixed crop-livestock systems in South Omo, southern Ethiopia

BackgroundIn the pastoral/agro-pastoral communities in Ethiopia, like in South Omo, brucellosis constitutes a serious health threat for livestock and the public. The public health risk is especially high in these communities, as their way of life is highly linked with their herds. ObjectiveThe study was conducted to estimate the seroprevalence and identify potential risk factors of cattle brucellosis in South Omo zone in southern Ethiopia. MethodsA total of 614 traditionally managed local zebu female cattle, above six months old, were bled and data on hypothesized risk factors were collected using a semi-structured questionnaire. The preliminary screening of the sera for Brucella antibodies was done using Rose Bengal Plate Test (RBPT) and positive sera were further subjected to complement fixation test (CFT). ResultsThe overall animal level seroprevalence of brucellosis was 2.8 % (95 % CI: 1.72–4.41) while herd level prevalence was 11.3 % (95 % CI: 6.5–19.0). Among the risk factors considered, seroprevalence was associated with herd size, new animal introduction, district, history of occurrence of abortion, and retained fetal membranes (RFM), at both individual- and herd-level (p < 0.05). Higher seroprevalence of brucellosis was observed in cows than heifers and in animals older than 4 years (p < 0.05). Brucella seroprevalence was higher in herds in lowland areas than those in mid-altitude and highlands (p < 0.05). ConclusionThe individual and herd level prevalence observed in our study indicates endemicity of brucellosis and the potential public health threat it poses in pastoral and agro-pastoral areas of southern Ethiopia. The results of the study also suggest that the disease might be responsible for significant losses in cattle productivity due to impaired reproductive performance.

Right Ventricular Strain Improves the Echocardiographic Diagnosis and Risk Stratification of Transthyretin Cardiac Amyloidosis Among Other Phenotypes of Left Ventricular Hypertrophy

Cardiac amyloidosis is a diffuse disease affecting all cardiac chambers. The value of right ventricular free-wall strain is uncertain as an echocardiographic red flag. We hypothesized that right ventricular free-wall strain is of added value for diagnostic and prognostic purposes in patients with transthyretin cardiac amyloidosis (ATTR-CA). A diagnosis of ATTR-CA required positive Tc-99m pyrophosphate scintigraphy and negative serum clonal dyscrasia. Patients with left ventricular (LV) hypertrophy (LVH; interventricular septal thickness ≥1.2cm) by echocardiography and negative pyrophosphate scintigraphy served as controls after exclusion of amyloid light-chain cardiac amyloidosis. Longitudinal strain was computed with speckle-tracking echocardiography. We studied 108 subjects with ATTR-CA and 106 controls with LVH, retrospectively. Right ventricular free-wall strain was independently associated with the diagnosis of ATTR-CA after adjusting for classical echocardiographic parameters, namely, relative apical sparing (RAS), e', and E/e'. Right ventricular free-wall strain≥-16% was incremental to LV RAS in the overall group and in the subgroup without extreme wall thickness (≤1.4cm; Harrell's C, net reclassification improvement=0.213, P<.001; and net reclassification improvement= 0.463, P=.015, respectively). Major adverse cardiovascular and cerebrovascular events (heart failure hospitalization, stroke, death) occurred in 47 ATTR-CA patients, during follow-up (median, 38; range, 6-60months). Right ventricular free-wall strain≥-16% was associated with 3-fold increased risk of MACCE in ATTR-CA patients independently of age, comorbidities, B-type natriuretic peptide, and tafamidis treatment. Right ventricular free-wall strain was additive to LV ejection fraction for risk stratification (chi square =10.2; P=.017). Right ventricular free-wall strain>-16% has incremental value to LV RAS for the differential diagnosis of ATTR-CA among LVH phenotypes and is associated with poor prognosis.

HIV rapid tests immunological internal control can be misleading

Objective: To assess whether internal controls (IC) of HIV rapid tests (HRT) and a rapid confirmatory assay (Geenius) were designed to solely prevent procedural missteps or wether they can also alert for insufficient sample quantity.Material and methods: Thirteen tests were evaluated using distilled water, phosphate buffered saline, and HIV-1 positive serum samples : Autotest VIH® (Autotest) (AAZ-LMB), INSTITM HIV-1/HIV-2 Antibody Test (INSTI) (Biolytical), MULTISURE® HIV Rapid Test (MULTISURE) (MP Diagnostics Asia Pacific Pte. CE Ltd), Biosynex® HIVTOP (Biosynex), Exacto HIVTOP® (Biosynex), Exacto® TEST HIV PRO (Biosynex), GenieTM Fast HIV 1/2 (Bio-Rad laboratories), GeeniusTM HIV 1/2 Confirmatory Assay (Geenius) (Bio-Rad laboratories), VIKIA® HIV 1/2 (bioMérieux), FIRST RESPONSE® Test VIH 1-2.O CARTE (Premier Medical Corporation), Hexagon HIV (Human), DetermineTM HIV EARLY DETECT (Determine) (Abbott), and ONE STEP Anti-HIV 1&2 (InTec PRODUCTS, Xiamen, Fujian P. R. China).Results: Among the 13 tests, 4 were invalid when tested without human serum sample: Autotest, Geenius, INSTI and MULTISURE. The Autotest and INSTI accurately labelled diluted samples, with a Test line positive at least one dilution below the IC. Geenius test line and IC were positive up to the same dilution with automated reading. However, MULTISURE IC was positive up to 1000-fold dilution higher than its test line, making it unsuitable to alert for low sample quantity.

Epitope mapping and establishment of a blocking ELISA for mAb targeting the p72 protein of African swine fever virus

The African swine fever virus (ASFV) has the ability to infect pigs and cause a highly contagious acute fever that can result in a mortality rate as high as 100%. Due to the viral epidemic, the pig industry worldwide has suffered significant financial setbacks. The absence of a proven vaccine for ASFV necessitates the development of a sensitive and reliable serological diagnostic method, enabling laboratories to effectively and expeditiously detect ASFV infection. In this study, four strains of monoclonal antibodies (mAbs) against p72, namely, 5A1, 4C4, 8A9, and 5E10, were generated through recombinant expression of p72, the main capsid protein of ASFV, and immunized mice with it. Epitope localization was performed by truncated overlapping polypeptides. The results indicate that 5A1 and 4C4 recognized the amino acid 20–39 aa, 8A9 and 5E10 are recognized at 263–282 aa, which is consistent with the reported 265–280 aa epitopes. Conserved analysis revealed 20–39 aa is a high conservation of the epitopes in the ASFV genotypes. Moreover, a blocking ELISA assay for detection ASFV antibody based on 4C4 monoclonal antibody was developed and assessed. The receiver-operating characteristic (ROC) was performed to identify the best threshold value using 87 negative and 67 positive samples. The established test exhibited an area under the curve (AUC) of 0.9997, with a 95% confidence interval ranging from 99.87 to 100%. Furthermore, the test achieved a diagnostic sensitivity of 100% (with a 95% confidence interval of 95.72 to 100%) and a specificity of 98.51% (with a 95% confidence interval of 92.02 to 99.92%) when the threshold was set at 41.97%. The inter- and intra-batch coefficient of variation were below 10%, demonstrating the exceptional repeatability of the method. This method can detect the positive standard serum at a dilution as high as 1:512. Subsequently, an exceptional blocking ELISA assay was established with high diagnostic sensitivity and specificity, providing a novel tool for detecting ASFV antibodies.Key points• Four strains of ASFV monoclonal antibodies against p72 were prepared and their epitopes were identified.• Blocking ELISA method was established based on monoclonal antibody 4C4 with an identified conservative epitope.• The established blocking ELISA method has a good effect on the detection of ASFV antibody.

No COVID-19 pandemic impact on incidence and clinical presentation of celiac disease in Italian children.

We aimed to evaluate the impact of Coronavirus Disease-19 (COVID-19) pandemic on the incidence and clinical presentation of celiac disease (CD) in children. The diagnoses of CD were compared between the COVID-19 pandemic (from April 2020 to March 2022) and the pre-pandemic period (from April 2018 to March 2020) in three Italian Paediatric Gastroenterology centres (Varese, Como, Catanzaro). Electronic patient records were reviewed and additional information were collected through parental interview. The diagnosis of CD was made according to ESPGHAN criteria. SARS-CoV-2 infection was diagnosed based on pre-vaccination positive serum antibodies or nasopharyngeal swabs. Z test and chi-square were used for statistical analysis. The overall number of paediatric diagnosis of CD did not differ between the two years pre-pandemic and pandemic periods (177 and 172 cases) in the three Italian participating centres. Clinical presentation of CD was also similar throughout the study periods. SARS-CoV-2 infection has been documented in 10.6% of children but only in 5.8% of these occurred before CD diagnosis. Different to what reported for other autoimmune diseases, the incidence and presenting symptoms of CD in our paediatric population did not change during the COVID-19 pandemic compared to the previous 2 years.

#2560 Intradialytic serum phosphate variations are associated with low PTH levels

Abstract Background and Aims Secondary hyperparathyroidism (SHPT) is one of the most common complications in patients with chronic kidney disease (CKD). The prevalence of SHPT progressively increases with the severity of CKD, affecting approximately 50-70% of patients on hemodialysis (HD). SHPT significantly impacts morbidity, mortality, hospitalization risks, healthcare costs, and the quality of life of CKD patients, making its management of paramount importance in this population. A positive serum phosphorus (P) balance is one of the critical determinants of SHPT, and the management of P levels in HD patients revolves around nutritional therapy, medications, and optimal kidney replacement therapy (KRT). Numerous studies have explored the role of KRT in P control among prevalent HD patients. However, whether the reduction of P achieved during KRT affects parathyroid hormone (PTH) levels is still a matter of debate. Currently, a direct evaluation of the relationship between intra-HD P reduction and PTH levels is lacking. Method We conducted a retrospective observational study on the prevalent HD population at the Division of Nephrology, University Hospital of Verona, from January to December 2022. We Included clinically stable adult patients undergoing HD for over 6 months, with multiple recorded visits during the follow-up. Demographic, clinical, laboratory, and medication data were collected. Time-varying variables were updated at each study visit. Food intake was assessed through a 3-day food record (HD day, non-HD day, and weekend day). Patients were instructed to maintain their usual ideally low-phosphate diet and to meticulously record and weigh all foods and preparations consumed. This assessment was conducted every 4 months during the observation period. From the 3-day food records, the dietary phosphorus intake was calculated using a national database and reported as mg of P per day. A dietitian evaluated dietary questionnaires. The primary outcome of interest was PTH levels. The absolute intra-HD change in P (intra-HD ∆P), defined as the difference between pre- and post-HD P levels, served as the main exposure. Multivariable adjusted linear mixed models were used to investigate the relationship between intra-HD ∆P and PTH levels. Potential confounders were included in the following three models of hierarchical adjustment: (i) Model 1, unadjusted; (ii) Model 2, adjusted for age and sex; (iii) Model 3, adjusted for all covariates in Model 2 plus URR, HD modality, active vitamin D, phosphate binders, calcimimetics, comorbidities. Since the relationship between intra-HD ∆P and PTH is influenced by the pre-HD P, all models were adjusted for pre-HD P. The association between PTH and intra-HD ∆P was also assessed in a subgroup of patients who provided a valid dietary diary. A multivariable linear mixed model adjusted for all covariates in Model 3 plus dietary phosphorus intake was used to assess this association. Results A total of 211 patients contributed to 904 study visits. A significant and positive relationship was observed between intra-HD ∆P and pre-HD P (β = 0.76, 95% CI 0.75, 0.78, p &amp;lt; 0.001) and urea reduction ratio (β = 0.38, 95% CI 0.35, 0.41; p &amp;lt; 0.001) (Figure). An increase in intra-HD ∆P was significantly and independently associated with low PTH levels (β = −0.17, 95% CI −0.30, −0.03; p = 0.016; Table). Overall, 86 out of 211 patients provided valid dietary data. The relationship between intra-HD ∆P and PTH levels was confirmed after additional adjustment for dietary P intake (p = 0.041). Conclusion The extent of intra-HD P reduction significantly correlates with low PTH levels. Strategies focused on optimizing or enhancing depurative efficiency in KRT can exert a substantial impact on managing positive phosphorus balance and secondary hyperparathyroidism (SHPT). The assessment of intra-HD P reduction may play a pivotal role in the management and follow-up of SHPT in HD patients.

#1862 Post-kidney transplantation de novo monoclonal gammopathy of undetermined significance—impact on patients and graft long-term outcomes

Abstract Background and Aims Monoclonal gammopathy of undetermined significance (MGUS) is an asymptomatic premalignant plasma cell condition with a reported prevalence of 3 to 4% in population older than 50 years and progression to multiple myeloma in approximately 1% of the cases. The clinical significance of MGUS in patients diagnosed prior to kidney transplantation (KT) has previously been studied, however, the impact on survival and morbidity of newly diagnosed MGUS on the period after KT, remains unclear. The aim of this study was to compare the outcomes between patients with MGUS emerging after KT versus non-MGUS patients. Method This is a retrospective single-center cohort analysis of a population of 793 kidney transplant recipients between 1985 and 2022. Diagnosis of MGUS after KT was based on positive serum protein electrophoresis and confirmed by serum immunofixation. We performed a randomized selection of a matched control group, adjusted for age, gender, and KT duration. Both groups were compared for number of infections (bacterial and viral), acute rejections, neoplastic complications, and for overall patient and graft survival. Results This study included a total of 112 patients, with a mean age of 60 years (±11), of which 86 (77%) were older than 50 years. Median follow-up duration was 149 [IQR 89-207] months. We identified 56 patients (7%) with MGUS after KT. Only 1 (1.7%) patient progressed to multiple myeloma. Induction immunosuppression type was not associated with higher development of MGUS. We found no difference between the MGUS group and the control group in the incidence of infections, acute graft rejections, or solid and hematologic neoplasia (Table 1). There was also no difference between overall patient and graft survival. Conclusion MGUS is a premalignant disease frequently present in KT recipients, with a higher prevalence compared to the general population, although its development after KT does not appear to contribute to a worse prognosis or to a higher progression to multiple myeloma.

Epidemiological analysis of the current prevalence of hepatitis B virus infection among pregnant and postpartum women in China from 2021 to 2023

Objective: To analyze hepatitis B serologic tests and the current prevalence of hepatitis B virus (HBV) infection among pregnant and postpartum women in China from 2021 to 2023. Methods: Data on managing the prevention of mother-to-child transmission of HIV, syphilis, and hepatitis were retrieved from the National Information System. A positive serum HBsAg test was used to define HBV infection. The χ(2) test was used to compare the coverage rate of the hepatitis B serologic test across different years, in early-stage pregnancy, and the current HBV infection in pregnant and postpartum women. A two-sided P value of <0.05 was considered a statistically significant difference. Results: The coverage rate for hepatitis B serological detection in pregnant (including intrapartum) and postpartum women and early-stage pregnancy rose from 99.68% (10 463 059/10 496 883) and 82.96% (8 707 765/10 496 883) to 99.94% (8 678 777/8 684 387, P < 0.001) and 88.87% (7 717 857/8 684 387, P < 0.001) in China between 2021 and 2023. The current prevalence rate of HBV infection decreased from 4.98% (521 479/10 463 059) in 2021 to 4.56% (396 148/8 678 777) in 2023 among pregnant and postpartum women (P < 0.001). The current prevalence rate of HBV infection ranged from 1.53% to 10.39% among pregnant and postpartum women in various provinces of China in 2023. Conclusion: The coverage rate for hepatitis B serologic tests in China increased significantly between 2021 and 2023 in pregnant and postpartum women. Therefore, the current prevalence rate of HBV infection has decreased significantly in pregnant and postpartum women, but a regional difference still exists.

Anti-tubulin beta 4A (TBB4A) antibody immobilized sperm in a complement-dependent manner in humans

Sperm-immobilizing antibodies (SI-Abs) are detected in the sera of 3 % of infertile women. SI-Abs are occasionally produced as allogeneic antibodies against sperm, causing immune infertility. SI-Abs inhibit the passage of sperm through the female reproductive tract. Research on anti-sperm antibodies (ASA) remains of great importance for population control. We aimed to identify the antigens recognized by SI-Abs and elucidate the pathogenesis of immune infertility. Twelve sperm-immobilization test (SIT)-positive and fourteen SIT-negative sera were analyzed by two-dimensional electrophoresis and western blotting. Antigenic materials were extracted from well-motile sperm prepared using 0.1 % sodium dodecyl sulfate. In total, 22 different spots were detected in the 12 positive sera. Among these, three positive serum samples showed two positive signals with similar migration patterns. The significant positive spots were Mr: 49 K, pI: 5.1 and Mr: 51 K, pI: 5.6. All these positive spots were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS); tubulin beta-4A (TBB4A) was identified from the spot Mr: 49 K, pI: 5.1. TBB4A is a major component of tubulin and constitutes the axoneme in the sperm tail and the centrosome in the sperm neck; it is generally located inside the cell. An authentic antibody against TBB4A showed a positive reaction in the sperm neck and tail regions in an immunofluorescence study. This antibody also inhibited sperm motility in a complement-dependent manner. Sperm membrane permeability reportedly changes during swimming and capacitation. We identified TBB4A as an antigenic molecule recognized by SI-Abs, which may be relevant to immunological contraception in the future.

A Study to Determine the Reason for Lower Pregnancy Rates in Younger Women with Diminished Oocyte Reserve-less Chance of Implanting vs. Fetal Demise

Most studies find lower live-delivered pregnancy rates (LDPRs) following in vitro fertilization-embryo transfer (IVF-ET) in women with diminished oocyte reserve (DOR) vs. normal oocyte reserve (NOR) even in a younger population. How much of a discrepancy may depend on the degree of oocyte depletion in the DOR group and the follicular stimulation protocol. Some fertility specialists favor an FSH receptor up-regulation technique as the protocol to attain the maximum LDPRs in women with DOR. The objective of this study was to compare chemical, clinical, and LDPRs following IVF-ET to determine if the main time of embryo loss is very early, as evidenced by the largest discrepancy occurring in attaining even a chemical pregnancy, and/ or a large discrepancy between a chemical pregnancy and attaining a clinical pregnancy (ultrasound evidence of a gestational sac) or later losses as evidenced by showing a greater loss rate from clinical evidence of pregnancy to live delivery in those with DOR compared to NOR. Overall, the DOR group, with a mean serum anti-Mullerian hormone (AMH) level of 0.42 ng/mL, had 50% as much chance to have an LDPR/transfer as women with NOR (AMH of 4.66) despite the same number of day 3 embryos transferred. The main reduction in LDPRs occurred from embryo transfer failing to attain a positive clinical pregnancy in the DOR group. The least discrepancy was from attaining a clinical pregnancy to live delivery. Thus, for NOR from positive pregnancy test 59% of this younger age group will have a live delivery vs. 50% for DOR. Thus, the reduction in LDPRS/transfer in young women with DOR vs. NOR seems mostly very early so the DOR group does not even attain a positive serum beta human chorionic gonadotropin level. This suggests that this inferiority in attaining a live delivery may be related to aneuploidy involving large chromosomes or a marked decrease in the mitochondrial DNA of the embryo.

Age-Related Decrease in Pellino-1 Expression Contributes to Osteoclast-Mediated Bone Loss.

Aging-related bone loss is driven by various biological factors, such as imbalanced bone metabolism from decreased osteoblast and increased osteoclast activities. Various transcriptional and post-transcriptional factors increase osteoclast activity with aging; however, studies regarding the post-translational regulators of osteoclast activity are still limited. The ubiquitin E3 ligase Pellino-1 is a well-known post-translational regulator of inflammation. However, how Pellino-1 expression regulation affects osteoclast differentiation remains unclear. This study determined that Pellino-1 levels are reduced in bone marrow monocytes (BMMs) from 40-week-old mice compared to 4-week-old mice. Interestingly, conditional Knockout (cKO) of Pellino-1 in 6-week-old mice resulted in decreased bone mass, reduced body size, and lower weight than in Pellino-1 floxed mice; however, these differences are not observed in 20-week-old mice. The increased number of tartrate-resistant acid phosphatase (TRAP)-positive cells and serum levels of C-terminal telopeptides of type I collagen, a marker of bone resorption, in 6-week-old Pellino-1 cKO mice implied a connection between Pellino-1 and the osteoclast population. Enhanced TRAP activity and upregulation of osteoclast genes in BMMs from the cKO mice indicate that Pellino-1 deletion affects osteoclast differentiation, leading to decreased bone mass and heightened osteoclast activity. Thus, targeting Pellino-1 could be a potential gene therapy for managing and preventing osteoporosis.

Two Cases of Pediatric Leucine-Rich Glioma-Inactivated Protein-1 Encephalitis: Clinical Course, Challenges, and Implications

BackgroundLeucine-rich glioma-inactivated protein 1 (LGI-1) encephalitis is a rare form of autoimmune limbic encephalitis. Although relatively well documented in adults, pediatric cases are rare and remain poorly understood. MethodsWe reviewed two pediatric cases of LGI-1 encephalitis from a single tertiary care facility retrospectively. The detailed analysis included assessment of the initial presentation, clinical progression, diagnostic challenges, treatments, and outcome. To contextualize the differences between pediatric and adult manifestations of disease, we compared these findings with existing literature. ResultsBoth cases illustrate the diagnostic challenges faced at initial presentation due to the rarity of this diagnosis in children and the absence of characteristic faciobrachial dystonic seizures, which is common in adults. The constellation of neuropsychiatric symptoms and refractory focal seizures led to a high clinical suspicion for autoimmune encephalitis, therefore, both cases were treated empirically with intravenous methylprednisolone. The diagnosis in both cases was confirmed with positive serum antibody testing, reinforcing that LGI-1 antibodies are more sensitive in the serum rather than the cerebrospinal fluid (CSF). Seizure control and improvement in cognitive symptoms was achieved through a combination of immunotherapy and antiseizure medications. ConclusionsThis case series underscores the significance of considering LGI-1 encephalitis in the differential diagnosis of pediatric patients exhibiting unexplained neuropsychiatric symptoms and focal seizures and emphasizes the importance of performing both serum and CSF antibody testing. It is necessary to conduct further research to identify the full range of pediatric presentations and to determine the optimal treatment protocol.

Rapid Diagnosis of Pneumocystis jirovecii Pneumonia and Respiratory Tract Colonization by Next-Generation Sequencing

ObjectivesTo describe the epidemiology of Pneumocystis jirovecii pneumonia and colonization diagnosed by next-generation sequencing (NGS) and explore the usefulness of the number of P. jirovecii sequence reads for the diagnosis of P. jirovecii pneumonia.MethodsWe examined the NGS results for P. jirovecii in respiratory samples collected from patients and analysed their clinical, radiological and microbiological characteristics.ResultsAmong 285 respiratory samples collected over a 12-month period (January to December 2022), P. jirovecii sequences were detected in 56 samples from 53 patients. Fifty (94.3%) of the 53 patients were HIV-negative. Following our case definitions, 37 (69.8%) and 16 (30.2%) of the 53 patients had P. jirovecii infection and colonization respectively. P. jirovecii infection was associated with presence of underlying disease with immunosuppression (94.6% vs 18.8%, P < 0.05), positive serum 1,3-β-D-glucan (41.2% vs 0%, P < 0.01) and higher number of P. jirovecii sequence reads (P < 0.005). In contrast, P. jirovecii colonization was associated with the male sex (93.8% vs 54.1%, P < 0.01), another definitive infectious disease diagnosis of the respiratory tract (43.8% vs 2.7%, P < 0.001) and higher survival (100% vs 67.6%, P < 0.01). Although P. jirovecii pneumonia was associated with higher number of P. jirovecii reads in respiratory samples, only a sensitivity of 82.14% and a specificity of 68.75% could be achieved.ConclusionDetection of P. jirovecii sequences in respiratory samples has to be interpreted discreetly. A combination of clinical, radiological and laboratory findings is still the most crucial in determining whether a particular case is genuine P. jirovecii pneumonia.

PSV-18 Improved performance of PRRSV challenged pigs fed 1-α-glycerol monolaurate

Abstract Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Enteric Diarrhea (PEDV) has posed a persistent challenge to the swine industry despite vaccinations and biosecurity measures. Feed has been shown to be a carrier for bacterial and viral pathogens (Dee et al., 2020). Feed mitigation using medium chain fatty acids can eliminate the pathogen or reduce the pathogen load below its infectious dose. However, less is known regarding the application of monoglycerides for feed mitigation. The objective of this study was to determine if using only 1-α-glycerol monolaurate (GML) can be an effective feed mitigant against PRRSV compared with untreated control feed. A total of 2 rooms, each with 100 pigs per room, and 6 pens per room were used. The initial body weight of the pigs averaged 15 kg and were sourced from a herd free of PRRSV and PEDV. GML was added to the feed at 2 kg/mT. The study period was 14 d with starting and end body weights measured. An ice block viral challenge as described by Dee et al. (2020) was used. Briefly, a 500 mL ice cube containing 1x107 50% tissue culture infectious dose of each virus [PRRSV, PEDV and Senecavirus A (SVA)] was added to 1,300 kg of feed on d 0 and again on d 6. On d 15 pigs were tested for PRRSV (serum), PEDV (rectal swabs) and SVA (tonsil samples). Control and GML fed pigs did not seroconvert to PEDV and SVA, therefore, only results for PRRSV are presented. Average daily gain was significantly improved in GML fed pigs (0.50 kg/d vs 0.20 kg/d; P &amp;lt; 0.05). Percentage of PRRSV positive feed samples were decreased (P &amp;lt; 0.05) in GML-treated feed from 100% (Ct = 32.2) in control feed to 33% (Ct = 34.3). The percentage of positive oral fluid samples decreased (P &amp;lt; 0.05) from 100% (Ct = 26.1) in control pigs to 17.0% (Ct = 33.0) in GML fed pigs. GML fed pigs exhibited no clinical signs of PRRS (dysponea), whereas 100% of the control pigs had clinical signs (P &amp;lt; 0.05). Similarly, GML fed pigs had no PRRSV positive serum samples but 100% of the control pigs had PRRSV positive serum samples. Results from this study show that the use of only GML can be an effective feed mitigant to reduce or eliminant the clinical signs of PRRS in pigs and maintain growth performance.