Serological diagnostic for SARS-CoV-2: an experimental External Quality Assessment Scheme.

OBJECTIVES: Numerous analytical systems, rapidly made available on the market throughout the SARS-CoV-2 pandemic, aim to detect COVID-19, and to continuously update and improve the same systems. Medical laboratory professionals have also developed in-house analytical procedures in order to satisfy the enormous volume of requests for tests. These developments have highlighted the need control the analytical procedures used in order to guarantee patient safety. The External Quality Assessment (EQA) Scheme, an important quality assurance tool, aims to guarantee high standard performance for laboratory and analytical procedures. The aim of the present study was to report on the results collected in an experimental EQA scheme for the serological diagnosis of SARS-CoV-2. METHODS: All qualitative results collected in the different EQA surveys were summarized in order to identify the percentage of laboratory results in relation to typology of antibodies, results and samples. RESULTS: A total of 4,867 data sets were collected. The analysis of EQA data made, demonstrates a better agreement among laboratories results for total Ig than single immunoglobulins (IgG, IgM, IgA) in the case samples positive for SARS-CoV-2, and a wide divergence between IgM results for positive samples (only 34.9% were correct). Results for negative controls and specificity controls demonstrated a better overall agreement than results for positive samples. CONCLUSIONS: Working in collaboration with the IVD manufacturers, laboratory professionals must strive to achieve harmonization of results, and to develop well-defined protocols complying with the ISO 15189 requirements.

Disease monitoring with quantitative serum IgA levels provides a more reliable response assessment in multiple myeloma patients.

Unlike IgG monoclonal proteins (MCPs), IgA MCP quantification is unreliable due to beta-migration of IgA MCPs on serum protein electrophoresis (SPEP). The utility of nephelometric quantitative IgA (qIgA) to monitor IgA multiple myeloma (MM) is unclear. We retrospectively studied disease response kinetics using qIgA versus MCPs by SPEP, and developed and validated novel qIgA disease assessment criteria in 491 IgA MM patients. The SPEP MCP nadir occurred a median of 41 (IQR 0-102) days before the qIgA. The median time to achieve a partial response (PR) was shorter using standard IMWG versus qIgA response criteria (32 vs 58 days, p < 0.001). Stratification by qIgA criteria, unlike IMWG criteria, led to clear separation of the progression-free survival curves of patients achieving a PR or very good PR. There was a consistent trend toward earlier detection of disease progression using qIgA versus IMWG progression criteria. In conclusion, monitoring IgA MM using MCP-based IMWG criteria may be falsely reassuring, given that MCP levels on SPEP decrease faster than qIgA levels. The qIgA response criteria more accurately stratify patients based on the progression risk and may detect disease progression earlier, which may lead to more consistent measurement of trial endpoints and improved patient outcomes.

Quantitative assessment of successive carbohydrate additions to the clustered O-glycosylation sites of IgA1 by glycosyltransferases.

Mucin-type O-glycosylation occurs on many proteins that transit the Golgi apparatus. These glycans impact structure and function of many proteins and have important roles in cellular biosynthetic processes, signaling and differentiation. Although recent technological advances have enhanced our ability to profile glycosylation of glycoproteins, limitations in the understanding of the biosynthesis of these glycan structures remain. Some of these limitations stem from the difficulty to track the biosynthetic process of mucin-type O-glycosylation, especially when glycans occur in dense clusters in repeat regions of proteins, such as the mucins or immunoglobulin A1 (IgA1). Here, we describe a series of nano-liquid chromatography (LC)-mass spectrometry (MS) analyses that demonstrate the range of glycosyltransferase enzymatic activities involved in the biosynthesis of clustered O-glycans on IgA1. By utilizing nano-LC-MS relative quantitation of in vitro reaction products, our results provide unique insights into the biosynthesis of clustered IgA1 O-glycans. We have developed a workflow to determine glycoform-specific apparent rates of a human UDP-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltrasnfersase (GalNAc-T EC 2.4.1.41) and demonstrated how pre-existing glycans affect subsequent activity of glycosyltransferases, such as core 1 galactosyltransferase and α2,3- and α2,6-specific sialyltransferases, in successive additions in the biosynthesis of clustered O-glycans. In the context of IgA1, these results have potential to provide insight into the molecular mechanisms implicated in the pathogenesis of IgA nephropathy, an autoimmune renal disease involving aberrant IgA1 O-glycosylation. In a broader sense, these methods and workflows are applicable to the studies of the concerted and competing functions of other glycosyltransferases that initiate and extend mucin-type core 1 clustered O-glycosylation.

Effects of a work-related stress model based mental health promotion program on job stress, stress reactions and coping profiles of women workers: a control groups study.

BACKGROUND: Work-related stress and its detrimental effects on human health have rapidly increased during the past several years. It causes many different stress reactions, related diseases and unhealthy behavior among workers, but especially women workers. Thus, the aim of this study was to examine the effects of the work-related stress model based Workplace Mental Health Promotion Programme on the job stress, social support, reactions, salivary immunoglobulin A and Cortisol levels, work absenteeism, job performance and coping profiles of women workers. METHODS: This study had a "pre-test post-test non-equivalent control groups" design and included 70 women workers (35 in each study group) selected by randomized sampling from two factories. The programme was delivered as an intervention including 12 weeks of follow-up. Reminder messages, videos, and WhatsApp texts were used at the follow-up stage. The research measurements were; the assessment form, the Brief Job Stress Questionnaire, the Brief Coping Profile Scale, salivary ELISA kits, and a self-reported check-list. RESULTS: There were no differences in sociodemographic characteristics, general health or working conditions between the Intervention and control groups(p > .05). Three months after the intervention, there was a significant decrease in job stress(p ≤ .001), physical and mental reactions' scores(p ≤ .001) and work absenteeism(p < .05), and there was an increase in job performance(p < .05), social support(p ≤ .001) among the intervention group. The programme showed positive effects on coping profiles(p < .05). After the intervention salivary-cortisol and IgA levels showed a statistically significant decrease(p < .05). A majority of effect sizes were very large (ηp2 > .14). CONCLUSIONS: Work-ProMentH was found to be effective and useful in job stress management and promotion of effective coping profiles. It enables its users to holistically assess worker stress and to plan and examine intervention programmes via a systematic approach. There is a need for more empirical studies that may support the data of the present study, but it is thought that the intervention can be maintained for the long-term. We recommend that occupational health professionals at workplaces should consider using this model-based cost-effective intervention, which seems easy and practical to apply in real-life situations. TRIAL REGISTRATION: ISRCTN registration ID: ISRCTN14333710 (2020/10/03, retrospective registration).

Postnatal distribution of ZnO nanoparticles to the breast milk through oral route and their risk assessment for breastfed rat offsprings.

Various studies in rodents have shown that nanoparticles are transferred to the breast milk. Under the present study, lactating Wistar rats were repetitively gavaged 5, 25, and 50 mg/kg bw of zinc oxide nanoparticles (ZnO-NPs) and 50 mg kg-1 bw of bulk zinc oxide (bZnO) for 19 days after parturition. The results showed that ZnO-NPs were absorbed in the small intestine of dams and distributed to the liver. Furthermore, ZnO-NPs were distributed to the intestine and liver of rat pups through dam's milk. No significant change in body weight was observed in the dams treated with ZnO-NPs or bZnO and their offsprings as compared to the control group. The spleen weight significantly increased in the rat dams treated with 50 mg kg-1 of ZnO-NPs. ZnO-NPs were mostly excreted through feces. The levels of liver cytochrome P450 reductase and serum total antioxidant capacity significantly decreased in the rat dams treated with ZnO-NPs (50 mg kg-1) and their offsprings. The levels of serum cytokines (tumor necrosis factor-alpha and interleukin-1 beta) and liver injury marker enzymes (alanine aminotransferase and aspartate aminotransferase) significantly increased in the rat dams treated with ZnO-NPs (25 and 50 mg kg-1) and their offsprings. The level of immunoglobulin A secretion in the intestinal fluid of rat dams and their offsprings is significantly increased by increasing the dose of ZnO-NPs. Histopathology of intestine and liver of offsprings whose rat dams were treated with ZnO-NPs (50 mg kg-1) showed gross pathological changes. These results provide information for the safety evaluation of ZnO-NPs use during lactation. In conclusion, a dose-dependent postnatal transfer of ZnO-NPs is hazardous to the breastfed offsprings.

Psychological effects of exercise on community-dwelling older adults.

BACKGROUND: In recent years, there have been an increasing number of older adults who suffer from mental disorders globally. OBJECTIVE: The objective of this study was to examine the effect of an intervention that consisted of an exercise program to improve the mental health of community-dwelling older adults. PARTICIPANTS AND METHODS: The recruited participants of this study were community-dwelling older adults aged ≥60 years who participated in a comprehensive health promotion program in Kakogawa, Japan. Participants in the intervention group received an exercise program that was developed for older adults using Thera-Band. To measure participants' mental health status, a Japanese version of the short form of the Profile of Mood States (POMS-SF) was used. Stress markers were measured, such as salivary cortisol, alpha-amylase, and sIgA levels. All participants provided salivary samples and completed psychological questionnaires at baseline and 6-month follow-up. RESULTS: No significant differences were observed between the intervention and control groups with respect to POMS-SF score and salivary biomarker profile at baseline. After the intervention, the intervention group showed a significant decrease in the POMS-SF "fatigue" score and cortisol level. No significant changes were observed in the control group. CONCLUSION: Simultaneous changes in feelings of fatigue and cortisol levels were observed among subjects who had received the intervention of regular exercise. Further research is needed to investigate the effectiveness of exercise intervention in improving mental health among older adults.

Paradigm shift in activity assessment of IgA nephropathy - optimizing the next generation of diagnostic and therapeutic maneuvers via glycan targeting.

INTRODUCTION: IgA nephropathy (IgAN) is the most common glomerular disease and has a poor prognosis. Appropriate therapeutic strategies are not currently available due to the lack of information regarding IgAN pathogenesis and the absence of appropriate tools to assess disease activity in IgAN, a long-term chronic disease. However, recent evidence revealed that aberrantly glycosylated serum IgA1, mostly galactose-deficient IgA1 (Gd-IgA1) and immune complexes (ICs) with autoantibodies against glycan-containing epitopes on Gd-IgA1 are essential effector molecules. AREAS COVERED: Assessing disease activity by urinalysis/renal biopsy has some limitations, resulting in conflicts regarding the efficacy of possible IgAN-specific therapies. We summarize the characteristics and molecular basis of Gd-IgA1 and related ICs, their clinical application for activity assessment and early diagnosis, and discuss glycan as a potent target of therapeutic agents based on glycan engineering in IgAN. EXPERT OPINION: Recently, Gd-IgA1 and related ICs have shown clinical value for disease activity assessment and IgAN diagnosis. This suggests a paradigm shift in IgAN treatment thus allowing development of appropriate clinical trials of patients with IgAN stages and objective evaluation of the efficacy of future treatments. Early screening and diagnosis may increase therapeutic options, including quantitative regulation of nephritogenic Gd-IgA1 using therapeutic antibodies and selective depletion of Gd-IgA1-producing cells via glycan engineering.

Effects of synbiotic supplementation on breast milk levels of IgA, TGF-ß1, and TGF-ß2.

BACKGROUND: Effects of probiotics on the immunological composition of breast milk have been investigated in a few previous studies. OBJECTIVES: The aims of this study were to determine the effects of synbiotic (probiotic plus prebiotic) supplementation on immunoglobulin A (IgA), transforming growth factor ß1 (TGF-ß1), and transforming growth factor ß2 (TGF-ß2) levels of breast milk and on diarrhea incidence in infants. METHODS: In this randomized, double-blind, and placebo-controlled trial, we recruited 80 lactating mothers who were exclusively breastfeeding their 3-month-old infants. We randomly divided the mothers into 2 groups to receive a daily synbiotic supplement (n = 40) or a placebo (n = 40) for 30 days. Demographic and clinical data (ie, health status) were obtained through an interview. The IgA levels of breast milk were detected by nephelometry, and the levels of TGF-ß1 and TGF-ß2 were measured using a commercial Platinum ELISA kit. RESULTS: The breast milk IgA increased significantly from 0.41 ± 0.09 to 0.48 ± 0.15 g/L in the supplemented group (P = .018), while in the placebo group, no significant changes were observed. Although the breast milk TGF-ß1 levels did not change significantly, the TGF-ß2 levels of breast milk increased significantly from 270 ± 37.8 to 382 ± 43.7 pg/mL in the supplemented group (P = .043). Also, the incidence of diarrhea in infants decreased significantly in the supplemented group while no significant changes were observed in the placebo group after the experimental period. CONCLUSION: Synbiotic supplementation may have positive effects on the immune composition of breast milk and the reduction of diarrhea incidence in infants.

Antibodies against deamidated gliadin peptides and tissue transglutaminase for diagnosis of pediatric celiac disease.

OBJECTIVES: The aim of the present study was to evaluate diagnostic performance and actual costs in clinical practice of immumoglobulin (Ig)G/IgA deamidated gliadin peptide antibodies (DGP) as a complement to IgA antibodies against tissue transglutaminase (tTG) for the diagnosis of pediatric celiac disease (CD). METHODS: All of the consecutive patients younger than 18 years tested for tTG and/or DGP, who underwent duodenal biopsy because of suspected CD in Stockholm and Gothenburg, Sweden, from 2008 to 2010, were included. Medical records were reviewed. RESULTS: Of 537 children who underwent duodenal biopsy, 278 (52%) had CD. A total of 71 (13%) were younger than 2 years and 16 (4%) had IgA deficiency. Sensitivity and specificity for tTG were 94% and 86%, respectively. Corresponding values for DGP were 91% and 26%. Positive predictive values (PPV) were 88% for tTG and 51% for DGP. There were 148 children who were tTG-negative and DGP-positive, of which only 5% (8/148) had villous atrophy. Among children younger than 2 years with normal IgA, PPV was 96% (25/26) for tTG and 48% (24/50) for DGP. In 16 IgA-deficient children, 11 were DGP positive, of which 5 had CD (PPV 45%). Eight of 278 cases of CD would possibly have been missed without DGP. The cost of adding DGP and consequently more biopsies to be able to detect 8 extra cases of CD was [Euro sign]399,520 or [Euro sign]49,940 per case. CONCLUSIONS: For diagnosing CD, tTG is superior to DGP, even in children younger than 2 years. Combining tTG and DGP does not provide a better tradeoff between number of missed cases of CD, number of unnecessary duodenal biopsies, and cost than tTG alone.

IgA deficiency and the MHC: assessment of relative risk and microheterogeneity within the HLA A1 B8, DR3 (8.1) haplotype.

INTRODUCTION: Selective IgA deficiency (IgAD; serum IgA concentration of <0.07 g/l) is the most common primary immunodeficiency in Caucasians with an estimated prevalence of 1/600. The frequency of the extended major histocompatibility complex haplotype HLA A1, B8, DR3, DQ2 (the "8.1" haplotype) is increased among patients with IgAD. MATERIALS AND METHODS: We carried out a direct measurement of the relative risk of homozygosity of the 8.1 haplotype for IgA deficiency in a population-based sample of 117 B8, DR3 homozygous individuals. RESULTS AND DISCUSSION: IgA deficiency was found to be present in 2 of 117 (1.7%) of these subjects, a figure that is concordant with estimates of relative risk from large case-control studies in the Swedish population. These data are consistent with a multiplicative model for the 8.1 haplotype contribution to IgA deficiency and contrasts with prior studies, suggesting a much higher risk for 8.1 homozygosity. Using a dense single nucleotide polymorphism marker analysis of the MHC region in HLA B8, DR3, DQ2 homozygous individuals, we did not observe consistent differences between cases (n = 26) and controls (n = 24). Overall, our results do not support the hypothesis that IgA deficiency is associated with a distinct subgroup of 8.1 related haplotypes, but rather indicate that risk is conferred by the common 8.1 haplotype acting in multiplicative manner.

Assessment of pulmonary antibodies with induced sputum and bronchoalveolar lavage induced by nasal vaccination against Pseudomonas aeruginosa: a clinical phase I/II study.

BACKGROUND: Vaccination against Pseudomonas aeruginosa is a desirable albeit challenging strategy for prevention of airway infection in patients with cystic fibrosis. We assessed the immunogenicity of a nasal vaccine based on the outer membrane proteins F and I from Pseudomonas aeruginosa in the lower airways in a phase I/II clinical trial. METHODS: N = 12 healthy volunteers received 2 nasal vaccinations with an OprF-OprI gel as a primary and a systemic (n = 6) or a nasal booster vaccination (n = 6). Antibodies were assessed in induced sputum (IS), bronchoalveolar lavage (BAL), and in serum. RESULTS: OprF-OprI-specific IgG and IgA antibodies were found in both BAL and IS at comparable rates, but differed in the predominant isotype. IgA antibodies in IS did not correlate to the respective serum levels. Pulmonary antibodies were detectable in all vaccinees even 1 year after the vaccination. The systemic booster group had higher IgG levels in serum. However, the nasal booster group had the better long-term response with bronchial antibodies of both isotypes. CONCLUSION: The nasal OprF-OprI-vaccine induces a lasting antibody response at both, systemic and airway mucosal site. IS is a feasible method to non-invasively assess bronchial antibodies. A further optimization of the vaccination schedule is warranted.

Quantification of cortisol, cortisol immunoreactive metabolites, and immunoglobulin A in serum, saliva, urine, and feces for noninvasive assessment of stress in reindeer.

This study was designed to develop reliable methods for quantification of cortisol and cortisol immunoreactive metabolites (C-CIM) and immunoglobulin A (IgA) in reindeer serum, saliva, urine, and feces as tools for the objective noninvasive assessment of well-being and immunocompetence in reindeer. Although C-CIM was readily quantifiable by radioimmunoassay in serum, urine, and feces, the levels in saliva samples were low, rendering quantification unreliable. Whereas IgA concentrations were high in feces samples, they were much lower, albeit quantifiable, in serum and urine; the levels in saliva samples were too low for quantification with the use of an enzyme-linked immunosorbent assay that we developed. Further studies are in progress to validate the usefulness of fecal levels of C-CIM and IgA in the assessment of welfare in reindeer.

Sulphasalazine in ankylosing spondylitis. A radiological, clinical and laboratory assessment.

In a 12-month double-blind placebo-controlled trial, the effect of sulphasalazine was studied in 40 patients with ankylosing spondylitis. The treatment group showed significant improvement in pain, stiffness, sleep disturbance (p less than 0.05), finger/floor distance, erythrocyte sedimentation rate, C-reactive protein, orosomucoid and IgA levels (p less than 0.01). There was improvement in sleep disturbance (p less than 0.05), finger/floor distance and erythrocyte sedimentation rate (p less than 0.01) in the placebo group. Sulphasalazine did not retard radiological progression as measured either by plain X-ray or computerised tomographic scans. Multiple analysis of variance did not show a significant difference in disease activity indicators between the 2 groups.

Pathophysiology of rhinitis. 1. Assessment of the sources of protein in methacholine-induced nasal secretions.

Nasal provocation tests with normal saline and methacholine (MC) were performed in 25 atopic and 27 nonatopic subjects in an effort to assess the sources of protein in induced airway secretions. Nasal lavages obtained at baseline and after provocation were analyzed for albumin, total protein, secretory IgA (sIgA), and total IgA. Compared with baseline levels or saline provocation, MC provocation increased the secretion of albumin (p less than 0.025), total protein (p less than 0.001), sIgA (p less than 0.025), and total IgA (p less than 0.025), but did not significantly affect the relative proportions of albumin-to-total protein (albumin percent) or sIgA-to-total IgA (sIgA/total IgA ratio). Nasal pretreatment with atropine significantly inhibited MC-induced secretion of all 4 proteins, again without affecting the albumin percent or the sIgA/total IgA ratio. Because MC is known to stimulate atropine-inhibitable secretion of glandular products, these data suggest that sIgA and albumin may accompany glandular secretions. Immunohistochemical analyses of nasal turbinates confirmed that secretory component was found only on serous cells within submucous glands. Thus, it appears that cholinergic stimulation may regulate sIgA secretion and thereby participate in local nasal (and possibly respiratory tract) immunity.