Search Results (28)

Background/Objectives: Anti-p200 pemphigoid is a rare and likely underdiagnosed autoimmune blistering disorder. Laminin γ1 and laminin β4 have been implicated as potential target antigens in its pathogenesis. Recently, a novel indirect immunofluorescence assay targeting anti-laminin β4 antibodies has been developed, demonstrating high sensitivity and specificity, and offering a valuable tool for improved diagnosis. Methods: Of the 451 patients, 21 were selected for further laboratory analysis based on medical records. Sera from 10 patients, which showed a positive direct immunofluorescence (DIF) result and negative results in multiplex enzyme-linked immunosorbent assays (ELISAs) and/or mosaic six-parameter indirect immunofluorescence (IIF) for various autoimmune bullous diseases, were tested for the presence of anti-laminin β4 antibodies. Additionally, sera from 11 patients with positive DIF and positive ELISA for antibodies against BP180 and/or BP230 were analyzed. Results: Among the 10 patients with positive DIF and negative ELISA and/or mosaic six-parameter IIF, 6 sera were positive for anti-laminin β4 antibodies. These patients presented with atypical clinical features. In contrast, all 11 sera from patients with both positive DIF and positive ELISA for BP180 and/or BP230 were negative for anti-laminin β4 antibodies. Conclusions: In patients with a positive DIF result but negative ELISA and/or mosaic six-parameter IIF findings, testing for anti-laminin β4 antibodies should be considered. Furthermore, in cases presenting with atypical clinical features—such as acral distribution of lesions, intense pruritus, or erythematous–edematous plaques—the possibility of anti-p200 pemphigoid should be included in the differential diagnosis. Full article

Background: Antinuclear antibodies (ANAs) serve as crucial biomarkers for diagnosing systemic autoimmune diseases; however, their interpretation can be complex and may not always correlate with clinical symptoms. Methods: A comprehensive narrative review was conducted to evaluate the peer-reviewed literature published between 1961 and 2025. Databases, including PubMed and Scopus, were searched using combinations of controlled vocabulary and free-text terms relating to antinuclear antibodies and their clinical significance. The objective was to gather and synthesize information regarding the diagnostic utility and interpretation of ANA testing in routine medical practice. Discussion: The indirect immunofluorescence assay (IIF) on HEp-2 cells is established as the gold standard for detecting ANAs, facilitating the classification of various fluorescent patterns. While a positive ANA test can suggest autoimmune disorders, the presence and titre must be interpreted alongside clinical findings, as low titres often lack diagnostic significance. Findings indicate that titres higher than 1:160 may provide greater specificity in differentiating true positives from false positives in healthy individuals. The study also emphasizes the relevance of fluorescence patterns, with specific patterns linked to particular diseases, although many do not have strong clinical correlations. Moreover, certain autoantibodies demonstrate high specificity for diseases like systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Ultimately, while ANA testing is invaluable for diagnosing connective tissue diseases, healthcare providers must consider its limitations to avoid misdiagnosis and unnecessary treatment. Conclusions: ANA testing is a valuable tool in the diagnosis of connective tissue diseases, but its interpretation must be approached with caution. Clinical context remains crucial when evaluating ANA results to avoid misdiagnosis and overtreatment. This review is about the diagnostic aspects and clinical consequences of ANA testing, as well as highlighting both the diagnostic benefits and the potential limitations of this procedure in everyday clinical practice. The review fills a gap in the literature by integrating the diagnostic and clinical aspects of ANA testing, with a focus on real-world interpretation challenges. Full article

Background: Patients suffering from a new variant of endemic pemphigus foliaceus in El Bagre, Colombia, South America (El Bagre-EPF) produce autoantibodies (Abs) to different proteins in the skin (frustre form), as well as to those in other organs (Senear–Usher-like and systemic forms). Here, we hypothesize whether patients’ autoantibodies play a role in triggering epicardium and pericardium autoimmunity and pathogenicity. We based this hypothesis on knowing that these patients frequently show clinical symptoms of the chest and heart, and we hypothesize that the autoantibodies of this disease are the main contributors to the base of the pericardial conditions of these patients. Materials and Methods: A case-control study for testing the sera of patients affected by El Bagre-EPF (n = 45) and matched controls from the endemic area (n = 45) was conducted to evaluate reactivity with the pericardial tissue. Patients’ necropsies were tested by immunohistochemistry (IHC), in El Bagre-EPF patients (n = 7) and matched controls. Results: The sera from most El Bagre-EPF patients displayed polyclonal autoreactivity with both layers of the pericardium, i.e., fibrous pericardium and serous pericardium (mainly to cell junctions and sensory nerve formations), as well as with the neurovascular cell junction branches. Controls were negative (p < 0.1). These reactivities were detected by IIF, CM, and IHC using secondary Abs against total IgG, IgM, Kappa and lambda, C3C of the complement, fibrinogen, and albumin. Furthermore, Abs against MIZAP, ARVCF, desmoplakin I-II, and p0071 colocalized with the Abs of El Bagre-EPF (p < 0.1). Conclusions: Patients affected by El Bagre-EPF produce autoantibodies directed against molecules present in the cell junctions of the pericardium and adnexal structures. Further studies will focus on the clinical significance of these findings. Full article

In veterinary medicine, the diagnosis of peripheral neuropathies is currently performed using semithin sections or nerve fiber teasing from nerve biopsy. However, these methods actually fail to identify more specific length-dependent and somatosensitive neuropathies. In humans, skin punch biopsy is used to diagnose the latter, through the identification and count of intraepidermal nerve fibers (IENFs) crossing the dermal–epidermal junction, with indirect immunofluorescence (IIF). However, the current need for frozen samples for this technique limits its routine application in clinical practice. In this study, we set up an IIF protocol to identify IENFs in dogs’ skin punch biopsies. Six tests were performed on canine formalin-fixed paraffin-embedded (FFPE) 8 mm skin punches, using an antibody anti-PGP9.5, also known as ubiquitin carboxyl-terminal hydrolase-1. Three parameters were checked: (1) the effectiveness of the co-localization immunoreaction, (2) the thickness of sections, and (3) the magnification for image acquisition. The best IIF results in terms of the sharpness of fiber visualization and the possibility to count them were obtained with 10 µm sections, with a high-power field (×40), without co-localization for nuclei and epithelial structures. Reference data concerning the IENF density of different skin regions in healthy animals of different ages remain to be defined for future diagnostic applications. Full article

Background and Objectives: Autoimmune thyroid diseases are more prevalent in patients diagnosed with Sjögren disease (SD) than in the general population. SD and autoimmune thyroid diseases are two distinct yet interrelated autoimmune disorders. The objective of this study was to determine the frequency of autoimmune thyroiditis (AT), autoantibody relationships, and clinical features in patients with SD. Materials and Methods: The study included 525 patients. A retrospective evaluation was conducted on the demographic data, biochemical and serological tests, and pathological data of the patients. An anti-nuclear antibody (ANA) test was performed using the indirect immunofluorescence (IIF) method using HEp-2 (HEp-2000) cells as substrate. The Schirmer test and minor salivary gland biopsy were conducted on all patients. Results: AT was detected in 167 (31.8%) of 525 patients who participated in the study. The anti-nuclear antibody (ANA) test and anti-SS-A positivity rate were higher in the AT group (p value < 0.001 and 0.002 respectively). We found that the likelihood of developing AT increased as ANA titres increased. ANA positivity titres were found to be significant at 2+, 3+, and 4+ values (odd ratios 2.41, 3.40, and 4.21, respectively). Additionally, histological examination of salivary gland biopsies revealed a significantly higher prevalence of diffuse lymphocytic infiltration in the AT group. Conclusions: AT was present in 31% of patients with SD. The presence of ANA positivity, anti-SS-A positivity, and diffuse lymphocytic infiltration appears to exert an influence on the association between these two diseases. Full article

Background/Objectives: Systemic autoimmune rheumatic diseases (SARDs) pose diagnostic challenges, particularly in pediatric populations, due to their diverse presentations and overlapping symptoms. This study aimed to evaluate the diagnostic concordance between indirect immunofluorescence (IIF) at different dilution levels (1/80 and 1/640) and immunoblot findings for anti-centromere antibody (ACA) positivity. Additionally, the clinical significance of ACA positivity and its association with SARDs in pediatric patients was assessed. Methods: This retrospective, cross-sectional study included 58 pediatric patients evaluated for anti-nuclear antibody (ANA) testing at Ege University Hospital from 2019 to 2021. IIF was performed using HEp-20-10 cells and immunoblot testing was conducted to assess CENP-B reactivity. Statistical analyses included chi-square tests, correspondence analysis, and regression modeling to explore the relationship between IIF titers, immunoblot findings, and SARD diagnoses. Results: Among the patients, 62.1% were diagnosed with SARD. Higher IIF titers (≥1/640) were strongly associated with CENP-B 3+ immunoblot positivity, while lower titers (1/80 and 1/320) correlated with CENP-B 1+. Patients with IIF positivity at 1/80 were 15.89 times more likely to have SARD (p < 0.001). Correspondence analysis revealed significant associations between IIF dilution levels and immunoblot reactivity (χ² = 37.574, p < 0.000). Gender and age were not significant predictors of SARD positivity. Conclusions: This study highlights the diagnostic value of higher IIF dilution levels (≥1/640) in improving ACA detection and SARD diagnosis in pediatric patients. Incorporating complementary diagnostic tools, such as immunoblot testing, can enhance diagnostic accuracy. These findings support adopting higher IIF cutoff levels in clinical practice for pediatric populations. Full article

Dregs waste, a byproduct from green liquor clarification in the pulp industry, is increasingly generated as global cellulose production rises. This accumulation, along with its underutilization, presents environmental challenges and opportunities for reuse. The study focuses on the high alkali content in dregs, which can impact cement durability. The main objective is to analyze the effect of dregs on the alkali–silica reaction in mortars. Dregs were incorporated into mortar mixtures at 0%, 5%, 10%, and 15% proportions relative to cement mass, using six types of Brazilian cement and a blend with silica fume. The alkali–silica reactivity was assessed via the accelerated mortar prism test (ABNT NBR 15577-4:2018), with compressive strength tests and scanning electron microscopy evaluating structural integrity and microstructural changes. The results indicated that adding 5% dregs improved compressive strength in certain mortars, such as CP V-ARI RS, CP II-F, and CP IV. However, at 15% dregs, compressive strength significantly decreased, particularly in CP V with 10% silica fume. Cements with high pozzolanic content, such as CP IV and CP III, showed strong potential to inhibit ASR expansion. However, of the 28 mixtures analyzed, only four containing CP III had expansions within the limits set by standards. This study highlights the potential of incorporating dregs as a supplementary material in cement, promoting sustainability in the industry and reducing environmental impact. Full article

Autoimmune diseases of the liver and biliary tract require timely and accurate diagnosis. This study evaluates the D-tek panel (D-Tek, Mons, Belgium) of 10 immunodot antigens for its effectiveness in diagnosing autoimmune hepatitis (AIH) and primary biliary cholangitis (PBC). We retrospectively analysed serum samples from 111 patients who had undergone routine testing, including indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assays (ELISA), to confirm or exclude autoimmune liver or biliary tract disease. The panel tested for M2/nPDC, M2/OGDC-E2, M2/BCOADC-E2, M2/PDC-E2, gp210, sp100, LKM1, LC1, SLA, and F-actin antigens. Results showed that all positive IIF+ELISA results were confirmed by the immunodot panel, except for two samples from patients who had never been diagnosed with AIH. The immunodot test identified over 20 additional autoantibodies in samples initially negative by IIF, corroborated by laboratory imaging and medical history. The immunodot technique proved to be a quick, sensitive, and specific method with high overall accuracy. This study suggests that the immunodot technique may be an effective screening and confirmatory method for autoimmune liver diseases, potentially improving diagnostic efficiency and accuracy in clinical practice. Full article

Parietal cell autoantibodies (PCAs), which recognize the enzyme H+/K+-ATPase as a target, are considered to be a diagnostic marker of autoimmune gastritis and pernicious anemia; these conditions are characterized by the presence of corpus atrophic gastritis. Circulating PCAs can be detected using several analytical methods that are commonly available in the clinical laboratory. Traditionally, indirect immunofluorescence (IIF) on rodent or primate stomach tissue is used as a screening test for the detection of PCAs. However, IIF suffers from a high inter-observer variability and lacks standardization. In addition, like immunoblotting, results are expressed only in a qualitative or semi-quantitative manner. Based on the few available studies that are reviewed herein, quantitative enzyme-linked immunosorbent assays (ELISAs) and fluorescence enzyme immunoassays (FEIAs) using purified H+/K+-ATPase perform better than IIF in the detection of PCAs, displaying higher sensitivity and utility in monitoring the disease. In light of their higher diagnostic accuracy, these solid-phase methods should be preferred to IIF in the screening of autoimmune atrophic gastritis. The use of methods to detect antibodies versus a specific subunit of H+/K+-ATPase (α or β) is currently confined to the world of research. Further investigation is required to define the clinical utility of H+/K+-ATPase subunit detection. Full article

Existing secure data aggregation protocols are weaker to eliminate data redundancy and protect wireless sensor networks (WSNs). Only some existing approaches have solved this singular issue when aggregating data. However, there is a need for a multi-featured protocol to handle the multiple problems of data aggregation, such as energy efficiency, authentication, authorization, and maintaining the security of the network. Looking at the significant demand for multi-featured data aggregation protocol, we propose secure data aggregation using authentication and authorization (SDAAA) protocol to detect malicious attacks, particularly cyberattacks such as sybil and sinkhole, to extend network performance. These attacks are more complex to address through existing cryptographic protocols. The proposed SDAAA protocol comprises a node authorization algorithm that permits legitimate nodes to communicate within the network. This SDAAA protocol’s methods help improve the quality of service (QoS) parameters. Furthermore, we introduce a mathematical model to improve accuracy, energy efficiency, data freshness, authorization, and authentication. Finally, our protocol is tested in an intelligent healthcare WSN patient-monitoring application scenario and verified using an OMNET++ simulator. Based upon the results, we confirm that our proposed SDAAA protocol attains a throughput of 444 kbs, representing a 98% of data/network channel capacity rate; an energy consumption of 2.6 joules, representing 99% network energy efficiency; an effected network of 2.45, representing 99.5% achieved overall performance of the network; and time complexity of 0.08 s, representing 98.5% efficiency of the proposed SDAAA approach. By contrast, contending protocols such as SD, EEHA, HAS, IIF, and RHC have throughput ranges between 415–443, representing 85–90% of the data rate/channel capacity of the network; energy consumption in the range of 3.0–3.6 joules, representing 88–95% energy efficiency of the network; effected network range of 2.98, representing 72–89% improved overall performance of the network; and time complexity in the range of 0.20 s, representing 72–89% efficiency of the proposed SDAAA approach. Therefore, our proposed SDAAA protocol outperforms other known approaches, such as SD, EEHA, HAS, IIF, and RHC, designed for secure data aggregation in a similar environment. Full article

Nanocomposites serving as dual (bimodal) probes have great potential in the field of bio-imaging. Here, we developed a simple one-pot synthesis for the reproducible generation of new luminescent and magnetically active bimetallic nanocomposites. The developed one-pot synthesis was performed in a sequential manner and obeys the principles of green chemistry. Briefly, bovine serum albumin (BSA) was exploited to uptake Au (III) and Fe (II)/Fe (III) ions simultaneously. Then, Au (III) ions were transformed to luminescent Au nanoclusters embedded in BSA (AuNCs-BSA) and majority of Fe ions were bio-embedded into superparamagnetic iron oxide nanoparticles (SPIONs) by the alkalization of the reaction medium. The resulting nanocomposites, AuNCs-BSA-SPIONs, represent a bimodal nanoprobe. Scanning transmission electron microscopy (STEM) imaging visualized nanostructures with sizes in units of nanometres that were arranged into aggregates. Mössbauer spectroscopy gave direct evidence regarding SPION presence. The potential applicability of these bimodal nanoprobes was verified by the measurement of their luminescent features as well as magnetic resonance (MR) imaging and relaxometry. It appears that these magneto-luminescent nanocomposites were able to compete with commercial MRI contrast agents as MR displays the beneficial property of bright luminescence of around 656 nm (fluorescence quantum yield of 6.2 ± 0.2%). The biocompatibility of the AuNCs-BSA-SPIONs nanocomposite has been tested and its long-term stability validated. Full article

The rapidly growing requirement for data has put forward Compressed Sensing (CS) to realize low-ratio sampling and to reconstruct complete signals. With the intensive development of Deep Neural Network (DNN) methods, performance in image reconstruction from CS measurements is constantly increasing. Currently, many network structures pay less attention to the relevance of before- and after-stage results and fail to make full use of relevant information in the compressed domain to achieve interblock information fusion and a great receptive field. Additionally, due to multiple resamplings and several forced compressions of information flow, information loss and network structure redundancy inevitably result. Therefore, an Information Enhancement and Fusion Network for CS reconstruction (IEF-CSNET) is proposed in this work, and a Compressed Information Extension (CIE) module is designed to fuse the compressed information in the compressed domain and greatly expand the receptive field. The Error Comprehensive Consideration Enhancement (ECCE) module enhances the error image by incorporating the previous recovered error so that the interlink among the iterations can be utilized for better recovery. In addition, an Iterative Information Flow Enhancement (IIFE) module is further proposed to complete the progressive recovery with loss-less information transmission during the iteration. In summary, the proposed method achieves the best effect, exhibits high robustness at this stage, with the peak signal-to-noise ratio (PSNR) improved by 0.59 dB on average under all test sets and sampling rates, and presents a greatly improved speed compared with the best algorithm. Full article

(1) Background: Myositis specific antibodies (MSA) are important diagnostic biomarkers. Among the rarest and most challenging MSA are anti-OJ antibodies which are associated with anti-synthetase syndrome (ASS). In contrast to the other tRNA synthetases that are targets of ASS autoantibodies (e.g Jo-1, PL-7, PL-12, EJ, KS, Zo), OJ represents a macromolecular complex with several ribonucleoprotein subunits. Therefore, the choice of the antigen in autoantibody assays can be challenging. (2) Methods: We collected two independent cohorts with anti-OJ antibodies, one based on a commercial line immunoassay (LIA) (n = 39), the second based on protein immunoprecipitation (IP) (n = 15). Samples were tested using a particle-based multi-analyte technology (PMAT) system that allows for the simultaneous detection of antibodies to various autoantigens. For the detection of anti-OJ antibodies, two different antigens were deployed (KARS, IARS) on PMAT. The reactivity to the two antigens KARS and IARS was analyzed individually and combined in a score (sum of the median fluorescence intensities). (3) Results: In the cohort selection based on LIA, 3/39 (7.7%) samples were positive for anti-KARS and 7/39 (17.9%) for anti-IARS and 14/39 (35.9%) when the two antigens were combined. In contrast, in samples selected by IP the sensitivity of anti-KARS was higher: 6/15 (40.0%) samples were positive for anti-KARS, 4/15 (26.7%) for anti-IARS and 12/15 (80.0%) for the combination of the two antigens. 18/39 (46.2%) of the LIA samples generated a cytoplasmic IIF pattern (compatible with anti-synthetase antibodies), but there was no association with the antibody levels, neither with LIA nor with PMAT. (4) Conclusions: The combination of IARS and KARS might represent a promising approach for the detection of anti-OJ antibodies on a fully automated platform. Full article

The resolution of complex medical diagnoses using pattern recognition requires an artificial neural network-based expert system to automate autoimmune disease diagnosis in blood samples. This process is done using image-based computer-aided diagnosis (CAD) to reduce errors in the diagnosis process. This paper describes a Multistage Classification Scheme (MSCS), which uses antinuclear antibody (ANA) tests to identify and classify the existence of autoantibodies in the blood serum that bind to antigens found in the nuclei of mammalian cells. The MSCS classified HEp-2 cells into three stages by using Binary Tree (BT), Artificial Neural Network (ANN), and Support Vector Machine (SVM) as basic blocks. The Indirect Immunofluorescence (IIF) technique is used in the ANA test with Human Epithelial type-2 (HEp-2) cells as substrates. The efficiency of the proposed methodology is assessed using the dataset of ICPR 2016. The intermediate cells (IMC) and positive cells (PC) were separated in Stage 1 prior to preprocessing based on their total strength, and special preprocessing is applied to intermediate cells for improved output, and positive cells are subjected to mild preprocessing. The mean class accuracy (MCA) was 84.9% for intermediate cells and 95.8% for positive cells, although the carefully picked 24 features and SVM classifier were applied. ANN showed better performance by adjusting the weights using the SCGBP algorithm. So, the MCA is 88.4% and 97.1% for intermediate and positive cells, respectively. BT had an MCA of 95.3% for intermediate and 98.6% for positive. In Stage 2, the meta learners BT2, ANN2, and SVM2 were trained for an augmented feature set (24 + 3 results from base learners). Therefore, the performance of BT2, ANN2, and SV M2 was increased by 1.8%, 4.5%, and 4.1% as compared to Stage 1. In Stage 3, the final prediction was performed by majority voting among the results of the three meta learners to achieve 99.1% MCA. The proposed algorithm can be embedded into a CAD framework built for the ANA examination. The proposed model will improve operational efficiency, decrease medical expenses, expand accessibility to healthcare, and improve patient safety in the sector, enabling enterprises to lower unplanned downtime, develop new products or services, increase operational effectiveness, and enhance risk management. Full article

The laboratory diagnostics of primary biliary cholangitis (PBC) have substantially improved, thanks to innovative analytical opportunities, such as enzyme-linked immunosorbent assays (ELISA) and multiple immunodot liver profile tests, based on recombinant or purified antigens. This study aimed to identify the best diagnostic test combination to optimize PBC diagnosis. Between January 2014 and March 2017, 164 PBC patients were recruited at the hospitals of Parma, Modena, Reggio-Emilia, and Piacenza. Antinuclear antibodies (ANA) and anti-mitochondrial antibodies (AMA) were assayed by indirect immunofluorescence (IIF), ELISA, and immunodot assays (PBC Screen, MIT3, M2, gp210, and sp100). AMA-IIF resulted in 89.6% positive cases. Using multiple immunodot liver profiles, AMA-M2 sensitivity was 94.5%, while anti-gp210 and anti-sp100 antibodies were positive in 16.5% and 17.7% of patients, respectively. PBC screening yielded positive results in 94.5% of cases; MIT3, sp100, and gp210 were detected by individual ELISA test in 89.0%, 17.1%, and 18.9% of patients, respectively. The association of PBC screening with IIF-AMA improved the diagnostic sensitivity from 89.6% to 98.2% (p < 0.01). When multiple immunodot liver profile testing was integrated with AMA-IIF, the diagnostic sensitivity increased from 89.1% to 98.8% (p < 0.01). The combination of IIF with solid-phase methods significantly improved diagnostic efficacy in PBC patients. Full article