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11 pages, 538 KiB  
Article
Evaluation of the HIV-1 Rapid Recency Assay and Limiting Antigen Avidity Enzyme Immunoassay for HIV Infection Status Interpretation in Long-Term Diagnosed Individuals in Thailand
by Supaporn Suparak, Petai Unpol, Kanokwan Ngueanchanthong, Siriphailin Jomjunyoung, Wipawee Thanyacharern, Sirilada Pimpa Chisholm, Nitis Smanthong, Thitipong Yingyong and Pilailuk Akkapaiboon Okada
Diagnostics 2025, 15(4), 444; https://doi.org/10.3390/diagnostics15040444 - 12 Feb 2025
Viewed by 288
Abstract
Background/Objectives: Accurate surveillance of recent HIV infections is crucial for effective epidemic control and timely intervention. The Limited Antigen Avidity Enzyme Immunoassay (LAg-EIA) allows precise differentiation between recent and long-term HIV infections. To enhance accessibility, it has been developted into a point-of-care test, [...] Read more.
Background/Objectives: Accurate surveillance of recent HIV infections is crucial for effective epidemic control and timely intervention. The Limited Antigen Avidity Enzyme Immunoassay (LAg-EIA) allows precise differentiation between recent and long-term HIV infections. To enhance accessibility, it has been developted into a point-of-care test, the Asanté™ HIV-1 Rapid Recency® Assay (ARRA), a rapid immunoassay. This study evaluated the performance and false recent rates (FRRs) of the ARRA, interpreted both visually and via a strip reader, in comparison with the LAg-EIA. Methods: Plasma samples were collected from two groups: 634 long-term HIV-infected individuals, identified through routine diagnostic testing, who had not received antiretroviral therapy for over one year, and 224 individuals from high-risk populations. High-risk individuals, including pregnant women, female sex workers, and men who have sex with men, were selected based on behavioral and demographic risk factors. Concordance between the ARRA and LAg-EIA was assessed, and FRRs were calculated for both assays. McNemar’s test was used to evaluate agreement, while Spearman’s rho was applied to assess correlation between the two methods. Results: Visual interpretation of ARRA demonstrated perfect agreement with LAg-Avidity EIA results (FRR = 0.00%), while the strip reader misclassified two specimens as recent infections (FRR = 0.32%). McNemar’s test indicated no significant differences between the methods (p > 0.05). Moderate agreement (Spearman’s rho = 0.434) was observed between ARRA strip reader results and LAg-Avidity EIA optical density values. Among high-risk populations, ARRA misclassified one sample as recent, resulting in an inconsistency rate of 0.45%. Conclusions: This study highlights ARRA’s reliability in identifying long-term infections and its potential as a point-of-care tool. Its rapid results and ease of use make it a valuable asset for effective HIV surveillance, facilitating targeted epidemic monitoring and enhancing public health interventions. Full article
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
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15 pages, 599 KiB  
Article
Prevalence and Risk Factors of Occult HCV Infection in the Adult Population of Mexico City
by Luis Antonio Uribe-Noguez, María Erandhi Prieto-Torres, Luis Octavio Uribe-Noguez, José Antonio Mata-Marín, Carla Ileana Arroyo-Anduiza, Rebeca Paquentín-Jimenez, Alberto Chaparro-Sanchez, Wendy Guadalupe Vazquez-Gonzalez, Andrea Santos Coy-Arechavaleta, Ericka Nelly Pompa-Mera, Jesus Gaytán-Martínez, Julio Elias Alvarado-Yaah, Clara Esperanza Santacruz-Tinoco and Alicia Ocaña-Mondragón
Viruses 2025, 17(2), 236; https://doi.org/10.3390/v17020236 - 8 Feb 2025
Viewed by 619
Abstract
Occult HCV infection (OCI) is defined by the presence of HCV RNA in hepatocytes and/or peripheral blood mononuclear cells (PBMCs) without detectable HCV RNA or anti-HCV antibodies in plasma. OCI is underrecognized and may contribute to HCV transmission. This study estimated OCI prevalence [...] Read more.
Occult HCV infection (OCI) is defined by the presence of HCV RNA in hepatocytes and/or peripheral blood mononuclear cells (PBMCs) without detectable HCV RNA or anti-HCV antibodies in plasma. OCI is underrecognized and may contribute to HCV transmission. This study estimated OCI prevalence and associated risk factors in adults from Mexico City. Methods: A retrospective cross-sectional study was conducted, analyzing 507 general population volunteers. Demographic data and potential risk factors were collected via questionnaire. Anti-HCV detection was performed using two techniques: immunochromatographic rapid test and chemiluminescent microparticle immunoassay (CMIA). Nested PCR was employed to detect HCV RNA in plasma and PBMCs. Positive samples were genotyped through sequencing and phylogenetic analysis of the Core/E1 region. Results: Of 507 participants, four were anti-HCV positive. HCV RNA was found in PBMCs of 27 individuals, while plasma samples tested negative, indicating a 5.3% OCI prevalence. OCI was significantly associated with blood donation (p = 0.015), drug use (p = 0.019), particularly cocaine (p = 0.001), and endoscopy (p = 0.043). Genotypes 1b, 1a, 2b, 3a, and 2j were detected in OCI cases. Conclusions: OCI prevalence in Mexico City’s general population is notable, with significant links to blood donation, cocaine use, and endoscopy. Enhanced diagnostic strategies are crucial to detect OCI and mitigate HCV transmission. Full article
(This article belongs to the Special Issue Viral Hepatitis and Liver Diseases)
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14 pages, 492 KiB  
Article
Risk Factors and Hospitalizations Associated with Pediatric Adenovirus and Rotavirus Infections in Northern Lebanon
by Sara Khalife, Marwan Osman, Sara Moubayed, Issmat I. Kassem and Dima El Safadi
Medicina 2025, 61(2), 296; https://doi.org/10.3390/medicina61020296 - 8 Feb 2025
Viewed by 525
Abstract
Background and Objectives: Acute gastroenteritis (AGE) is a major contributor to pediatric morbidity and mortality worldwide. There is a scarcity of data on AGE in North Lebanon, a region profoundly affected by the Syrian refugee crisis and water sanitation issues. This study [...] Read more.
Background and Objectives: Acute gastroenteritis (AGE) is a major contributor to pediatric morbidity and mortality worldwide. There is a scarcity of data on AGE in North Lebanon, a region profoundly affected by the Syrian refugee crisis and water sanitation issues. This study examines the prevalence, risk factors, and seasonal variations in adenovirus and rotavirus infections in children with AGE in North Lebanon. Materials and Methods: A cross-sectional study was conducted from September 2022 to August 2023 on 400 children (1 month to 5 years old) with AGE that were admitted to pediatric departments of three private hospitals in North Lebanon. Stool samples were collected and tested using chromatographic immunoassays. Comprehensive demographic and clinical data were collected and analyzed. Results: Rotavirus was the most frequent viral agent (28%), followed by adenovirus (12.3%) and mixed infections (5.5%). Rotavirus vaccination demonstrated a significant protective effect, and lower infection rates were associated with breastfeeding and consumption of bottled water (p < 0.001). Higher infection rates correlated with lower levels of maternal education and household incomes (p < 0.001). Malnutrition significantly increased susceptibility to rotavirus infections (p < 0.001). Conclusions: This study emphasized the urgent need for targeted public health interventions in North Lebanon to mitigate the burden of rotavirus and adenovirus-induced acute gastroenteritis among children. Full article
(This article belongs to the Section Pediatrics)
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25 pages, 719 KiB  
Review
Diagnostic Assays for Avian Influenza Virus Surveillance and Monitoring in Poultry
by Shahan Azeem and Kyoung-Jin Yoon
Viruses 2025, 17(2), 228; https://doi.org/10.3390/v17020228 - 6 Feb 2025
Viewed by 577
Abstract
Diagnostic testing plays a key role in a surveillance program as diagnostic testing aims to accurately determine the infection or disease status of an individual animal. Diagnostic assays for AIV can be categorized into four broad types: tests for detecting the virus, its [...] Read more.
Diagnostic testing plays a key role in a surveillance program as diagnostic testing aims to accurately determine the infection or disease status of an individual animal. Diagnostic assays for AIV can be categorized into four broad types: tests for detecting the virus, its antigen, its genomic material, and antibodies to the virus. Virus characterization almost always follows virus detection. The present article surveys the current literature on the goals, principles, test performance, advantages, and disadvantages of these diagnostic assays. Virus isolation can be achieved using embryonating eggs or cell cultures in a lab setting. Virus antigens can be detected by antigen-capturing immunoassays or tissue immunoassays. Viral RNA can be detected by PCR-based assays (gel-based reverse transcription–polymerase chain reaction (RT-PCR), or probe or SYBR® Green-based real-time RT-PCR), loop-mediated isothermal amplification, in situ hybridization, and nucleic acid sequence-based amplification. Antibodies to AIV can be detected by ELISA, agar gel immunodiffusion, hemagglutination inhibition, and microneutralization. Avian influenza virus can be characterized by hemagglutination inhibition, neuraminidase inhibition, sequencing (dideoxynucleotide chain-termination sequencing, next-generation sequencing), genetic sequence-based pathotype prediction, and pathogenicity testing. Novel and variant AIVs can be recognized by DNA microarrays, electron microscopy, mass spectroscopy, and Biological Microelectromechanical Systems. A variety of diagnostic tests are employed in AIV surveillance and monitoring. The choice of their use depends on the goal of testing (fit for purpose), the time of testing during the disease, the assay target, the sample matrix, assay performance, and the advantages and disadvantages of the assay. The article concludes with authors’ perspective of the use of diagnostic assays in the surveillance and monitoring of AIV in poultry. Full article
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15 pages, 10872 KiB  
Article
Enhanced Sensitivity Mach–Zehnder Interferometer-Based Tapered-in-Tapered Fiber-Optic Biosensor for the Immunoassay of C-Reactive Protein
by Lei Xiao, Xinghong Chen, Xuejin Li, Jinghan Zhang, Yan Wang, Dongqing Li, Xueming Hong, Yonghong Shao and Yuzhi Chen
Biosensors 2025, 15(2), 90; https://doi.org/10.3390/bios15020090 - 6 Feb 2025
Viewed by 473
Abstract
A Mach–Zehnder interferometer-based tapered-in-tapered fiber-optic biosensor was introduced in this paper. By integrating a micro-tapered fiber into a single tapered fiber structure, the design enhances sensitivity, signal-to-noise ratio, and resolution capability, while reducing the length of the sensing fiber. Through simulation analysis, it [...] Read more.
A Mach–Zehnder interferometer-based tapered-in-tapered fiber-optic biosensor was introduced in this paper. By integrating a micro-tapered fiber into a single tapered fiber structure, the design enhances sensitivity, signal-to-noise ratio, and resolution capability, while reducing the length of the sensing fiber. Through simulation analysis, it was found that the tapered-in-tapered fiber significantly improved the refractive index detection sensitivity by exciting a stronger evanescent field effect. The experimental comparison between the tapered-in-tapered fiber and traditional tapered fiber showed a 1.7-fold increase in sensitivity, reaching 3266.78 nm/RIU within the refractive index range of 1.3326 to 1.3414. Furthermore, to expand its application prospects in the biomedical field, glutaraldehyde cross-linking technology was used to immobilize C-reactive protein (CRP) antibodies on the surface of the tapered-in-tapered fiber, successfully creating a biosensing platform for the specific recognition of CRP. The experimental results demonstrate that this novel biosensor can rapidly and accurately detect CRP molecules at different concentrations with a detection limit of 0.278 μg/mL, and that it exhibits good selectivity and repeatability. This tapered-in-tapered fiber-optic biosensor provides new insights into the development of high-performance fiber-optic immunosensors and shows broad application potential in immunology research and early disease diagnosis. Full article
(This article belongs to the Section Optical and Photonic Biosensors)
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20 pages, 4942 KiB  
Article
Universal First-Trimester Screening Biomarkers for Diagnosis of Preeclampsia and Placenta Accreta Spectrum
by Angelika V. Timofeeva, Ivan S. Fedorov, Alla M. Tarasova, Yuliya V. Sukhova, Vyacheslav G. Kolod’ko, Tatiana Yu. Ivanets and Gennady T. Sukhikh
Biomolecules 2025, 15(2), 228; https://doi.org/10.3390/biom15020228 - 4 Feb 2025
Viewed by 572
Abstract
Background: Disruptions in epigenetic mechanisms regulating placentation, particularly imbalances in the levels of small non-coding RNAs, contribute to various pregnancy complications, including preeclampsia (PE) and placenta accreta spectrum (PAS). Given that abnormal trophoblast differentiation, invasiveness, and angiogenesis—reduced in PE and excessive in PAS—are [...] Read more.
Background: Disruptions in epigenetic mechanisms regulating placentation, particularly imbalances in the levels of small non-coding RNAs, contribute to various pregnancy complications, including preeclampsia (PE) and placenta accreta spectrum (PAS). Given that abnormal trophoblast differentiation, invasiveness, and angiogenesis—reduced in PE and excessive in PAS—are central to the pathogenesis of these conditions, this study aimed to identify universal circulating piRNAs and their targets. Methods: Small RNA deep sequencing, quantitative reverse transcription combined with real-time polymerase chain reaction, magnetic bead-based multiplex immunoassay, ELISA, and Western blotting were employed to quantify circulating piRNAs and proteins in the blood serum of pregnant women during the 11th–14th weeks of gestation. Results: Statistically significant negative correlations were identified between PE- and PAS-associated piRNAs (hsa_piR_019122, hsa_piR_020497, hsa_piR_019949, and piR_019675) and several molecules, including Endoglin, IL-18, VEGF-A, VEGF-C, Angiopoietin-2, sFASL, HB-EGF, TGFα, and Clusterin. These molecules are involved in processes such as angiogenesis, inflammation, the epithelial–mesenchymal transition, cell proliferation, adhesion, and apoptosis. A first-trimester pregnancy screening algorithm was developed using logistic regression models based on Clusterin concentration and the levels of hsa_piR_020497, hsa_piR_019949, piR_019675, and hsa_piR_019122. Conclusions: The proposed screening tool for early pregnancy monitoring may enable the prediction of PE or PAS in the first trimester, allowing timely interventions to reduce maternal and perinatal morbidity and mortality. Full article
(This article belongs to the Section Molecular Medicine)
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16 pages, 2306 KiB  
Article
Mapping Inflammatory Markers in Cerebrospinal Fluid Following Aneurysmal Subarachnoid Hemorrhage: An Age- and Sex-Matched Analysis
by Katharina Sophie Seyfried, Benedikt Kremer, Catharina Conzen-Dilger, Michael Veldeman, Ulf Bertram, Christian Blume, Christian Andreas Mueller, Tianshu Bi, Kerstin Jütten, Hans Clusmann and Anke Höllig
Int. J. Mol. Sci. 2025, 26(3), 1302; https://doi.org/10.3390/ijms26031302 - 3 Feb 2025
Viewed by 391
Abstract
Despite extensive research on aneurysm treatment and neurocritical care, aneurysmal subarachnoid hemorrhage (SAH) is still a life-threatening disease, often leaving survivors with lasting neurological and cognitive impairments. Early brain injury (EBI) and delayed cerebral ischemia (DCI) are the main contributors to brain damage, [...] Read more.
Despite extensive research on aneurysm treatment and neurocritical care, aneurysmal subarachnoid hemorrhage (SAH) is still a life-threatening disease, often leaving survivors with lasting neurological and cognitive impairments. Early brain injury (EBI) and delayed cerebral ischemia (DCI) are the main contributors to brain damage, with neuroinflammation being a critical shared pathophysiological process. While numerous inflammatory markers and their temporal profiles in cerebrospinal fluid (CSF) have already been identified, comparisons with age- and sex-matched controls are limited. This study analyzed CSF from 17 SAH patients requiring an external ventricular drain (EVD) due to symptomatic hydrocephalus, sampled on days 4 and 10 post-ictus. An age- and sex-matched control group included 17 cerebrovascularly healthy patients requiring lumbar drains during aortic surgery. Chemokines and cytokines were quantified using immunoassays. Significantly elevated markers in SAH patients across both time points included MCP-1, CXCL-13, Eotaxin-1, CXCL-10, IL-8, and MIF. MIP-1α and MIP-1β showed significant differences at particular time points, indicating a distinct temporal profile for each parameter. These findings highlight neuroinflammation’s key role in intracranial and systemic pathophysiology following SAH, emphasizing its complexity and individual variability. Knowing demographic factors impact the specific manifestations of pathophysiological processes, the comparison with an age- and sex-matched control group is meaningful. Full article
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15 pages, 1611 KiB  
Article
Towards the Measurement of Acute-Phase Proteins in Saliva in Farm Conditions: Development and Validation of a Lateral Flow Assay for the Measurement of C-Reactive Protein in Pigs
by Fernando Tecles, Asta Tvarijonaviciute, Simone Cavalera, Fabio Di Nardo, Claudio Baggiani, José Joaquín Cerón, Antonio González-Bulnes, María Elena Goyena, Silvia Martínez-Subiela, Lorena Franco-Martínez and Laura Anfossi
Chemosensors 2025, 13(2), 44; https://doi.org/10.3390/chemosensors13020044 - 2 Feb 2025
Viewed by 534
Abstract
Point-of-care diagnostic tests, such as lateral-flow immunoassay (LFIA), have emerged as a fast diagnostic tool in both human and veterinary medicine. In this paper, a gold nanoparticle-based LFIA device was developed for the measurement of C-reactive protein (CRP) in porcine saliva, using a [...] Read more.
Point-of-care diagnostic tests, such as lateral-flow immunoassay (LFIA), have emerged as a fast diagnostic tool in both human and veterinary medicine. In this paper, a gold nanoparticle-based LFIA device was developed for the measurement of C-reactive protein (CRP) in porcine saliva, using a monoclonal anti-porcine CRP antibody. The dilution ratio for the saliva samples was optimized at 1:5 with an assay buffer. The reaction time was optimized to 20 min, since this provided a positive signal with high CRP concentration saliva samples, but a negative result with an assay buffer or samples with a low CRP concentration. Linear results were observed when two samples with a high CRP concentration were serially diluted. Also, a linear relationship was observed with a validated quantitative method. The assay was precise when samples with high CRP concentration were measured five times in a single assay run. No overlap was observed when samples from healthy and diseased animals were analyzed. The LFIA allowed the detection of high CRP concentrations in porcine saliva samples. The intensity of the result was proportional to the CRP concentration obtained with the quantitative method, allowing for the possible use of the test for semiquantitative purposes. Full article
(This article belongs to the Special Issue Rapid Point-of-Care Testing Technology and Application)
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12 pages, 4661 KiB  
Communication
Evaluation and Validation of Colloidal Gold Immunochromatographic Qualitative Testing Products for the Detection of Emamectin Benzoate, Isocarbophos, and Fipronil in Cowpea Samples
by Anning Song, Miao Wang, Yongxin She, Maojun Jin, Zhen Cao, A. M. Abd El-Aty and Jing Wang
Viewed by 474
Abstract
Pesticide residues still pose a risk to human health. With the rapid development of rapid testing technology, the levels of different types of pesticide residues in agricultural products can be identified in a shorter period; thus, the safety of food can be guaranteed. [...] Read more.
Pesticide residues still pose a risk to human health. With the rapid development of rapid testing technology, the levels of different types of pesticide residues in agricultural products can be identified in a shorter period; thus, the safety of food can be guaranteed. However, the effectiveness of commercially available testing products has yet to be evaluated. In this study, colloidal gold immunochromatographic qualitative testing products manufactured by 34 companies were tested for their assay performance on Emamectin Benzoate, Isocarbophos, and fipronil with standardized cowpea samples. The results indicated that most of the evaluated products were identified as having ‘passed’. Most pesticide residue rapid test immunoassay products can be considered ideal means for testing certain pesticide residues. However, further evaluation of pesticide residue rapid test immunoassay products is needed, as detection technologies are still developing. Full article
(This article belongs to the Special Issue Residue Detection and Safety Control of Food Chemical Contaminants)
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14 pages, 2405 KiB  
Article
A Dual Nano-Signal Probe-Based Electrochemical Immunosensor for the Simultaneous Detection of Two Biomarkers in Gastric Cancer
by Li-Ting Su, Zhen-Qing Yang, Hua-Ping Peng and Ai-Lin Liu
Biosensors 2025, 15(2), 80; https://doi.org/10.3390/bios15020080 - 31 Jan 2025
Viewed by 600
Abstract
Detecting multiple tumor markers is of great importance. It helps in early cancer detection, accurate diagnosis, and monitoring treatment. In this work, gold nanoparticles–toluidine blue–graphene oxide (AuNPs-TB–GO) and gold nanoparticles–carboxyl ferrocene–tungsten disulfide (AuNPs–FMC–WS2) nanocomposites were prepared for labeling Carcinoembryonic antigen (CEA) [...] Read more.
Detecting multiple tumor markers is of great importance. It helps in early cancer detection, accurate diagnosis, and monitoring treatment. In this work, gold nanoparticles–toluidine blue–graphene oxide (AuNPs-TB–GO) and gold nanoparticles–carboxyl ferrocene–tungsten disulfide (AuNPs–FMC–WS2) nanocomposites were prepared for labeling Carcinoembryonic antigen (CEA) antibody and Carbohydrate antigen 72–4 (CA72-4) antibody, respectively, and used as two kinds of probes with different electrochemical signals. With the excellent magnetic performance of biotin immune magnetic beads (IMBs), the biofunctional IMBs were firmly deposited on the magnetic glassy carbon electrode (MGCE) surface by applying a constant magnetic field, and then the CEA and CA72-4 antibody were immobilized on the IMBs by the avidin–biotin conjugation. The assay was based on the change in the detection peak current. Under the optimum experimental conditions, the linear range of detection of CEA is of the two-component immunosensor is from 0.01 to 120 ng/mL, with a low detection limit of 0.003 ng/mL, and the linear range of detection of CA72-4 is from 0.05 to 35 U/mL, with a detection limit of 0.016 U/mL. The results showed that the proposed immunosensor enabled simultaneous monitoring of CEA and CA72-4 and exhibited good reproducibility, excellent high selectivity, and sensitivity. In particular, the proposed multiplexed immunoassay approach does not require sophisticated fabrication and is well-suited for high-throughput biosensing and application to other areas. Full article
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19 pages, 4594 KiB  
Article
Microarray-Based Avidity Assay for Assessment of Thyroid Autoantibodies
by Elena Savvateeva, Vera Sokolova, Marina Yukina, Nurana Nuralieva, Elena Kulagina, Maxim Donnikov, Lyudmila Kovalenko, Maria Kazakova, Ekaterina Troshina and Dmitry Gryadunov
Diagnostics 2025, 15(3), 341; https://doi.org/10.3390/diagnostics15030341 - 31 Jan 2025
Viewed by 435
Abstract
Background/Objectives: The aim of this study was to evaluate the avidity of thyroid autoantibodies (Abs) in sera of patients with autoimmune thyroid disease (AITD) and thyroid autoantibody carriers without diagnosed AITD. Methods: A hydrogel microarray-based multiplex assay with the chaotrope destruction stage was [...] Read more.
Background/Objectives: The aim of this study was to evaluate the avidity of thyroid autoantibodies (Abs) in sera of patients with autoimmune thyroid disease (AITD) and thyroid autoantibody carriers without diagnosed AITD. Methods: A hydrogel microarray-based multiplex assay with the chaotrope destruction stage was developed to measure the avidity of thyroid disease-associated autoantibodies, including those targeting thyroperoxidase (TPO), thyroglobulin (Tg), and other minor antigens. Results: Evaluation of the assay in three independent cohorts of patients, totaling 266 individuals with and without AITD, demonstrated the heterogeneous avidity of autoantibodies to thyroid proteins. For the confirmation study, the median avidity index (AI) for AbTg was 29.9% in healthy autoantibody carriers, 52.6% for AITD patients, and 92.7% for type 1 diabetes (T1D) thyroid autoantibody carriers. The median AI for AbTPO was 39.9% in healthy carriers, 73.4% in AITD patients, 83.2% in T1D thyroid autoantibody carriers, and 98.5% in AITD patients with thyroid neoplasm. In patients with Hashimoto’s thyroiditis and known disease duration, changes in the avidity maturation of AbTPO over time were demonstrated. Conclusions: Longitudinal studies of TPO- and/or Tg-positive healthy individuals (with an interval of 1–2 years between visits) are needed to evaluate the maturation of autoantibody avidity during the asymptomatic phase and to assess the potential of autoantibody avidity as a prognostic marker for disease development. Full article
(This article belongs to the Special Issue Recent Advances in Clinical Laboratory Immunology)
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16 pages, 2795 KiB  
Article
Mitochondria-Derived Vesicles and Inflammatory Profiles of Adults with Long COVID Supplemented with Red Beetroot Juice: Secondary Analysis of a Randomized Controlled Trial
by Emanuele Marzetti, Hélio José Coelho-Júnior, Riccardo Calvani, Giulia Girolimetti, Riccardo Di Corato, Francesca Ciciarello, Vincenzo Galluzzo, Clara Di Mario, Barbara Tolusso, Luca Santoro, Ottavia Giampaoli, Alberta Tomassini, Walter Aureli, Matteo Tosato, Francesco Landi, Cecilia Bucci, Flora Guerra and Anna Picca
Int. J. Mol. Sci. 2025, 26(3), 1224; https://doi.org/10.3390/ijms26031224 - 30 Jan 2025
Viewed by 557
Abstract
In a recent clinical trial, beetroot juice supplementation for 14 days yielded positive effects on systemic inflammation in adults with long COVID. Here, we explored the relationship between circulating markers of mitochondrial quality and inflammation in adults with long COVID as well as [...] Read more.
In a recent clinical trial, beetroot juice supplementation for 14 days yielded positive effects on systemic inflammation in adults with long COVID. Here, we explored the relationship between circulating markers of mitochondrial quality and inflammation in adults with long COVID as well as the impact of beetroot administration on those markers. We conducted secondary analyses of a placebo-controlled randomized clinical trial testing beetroot juice supplementation as a remedy against long COVID. Analyses were conducted in 25 participants, 10 assigned to placebo (mean age: 40.2 ± 11.5 years, 60% women) and 15 allocated to beetroot juice (mean age: 38.3 ± 7.7 years, 53.3% women). Extracellular vesicles were purified from serum by ultracentrifugation and assayed for components of the electron transport chain and mitochondrial DNA (mtDNA) by Western blot and droplet digital polymerase chain reaction (ddPCR), respectively. Inflammatory markers and circulating cell-free mtDNA were quantified in serum through a multiplex immunoassay and ddPCR, respectively. Beetroot juice administration for 14 days decreased serum levels of interleukin (IL)-1β, IL-8, and tumor necrosis factor alpha, with no effects on circulating markers of mitochondrial quality control. Significant negative associations were observed between vesicular markers of mitochondrial quality control and the performance on the 6 min walk test and flow-mediated dilation irrespective of group allocation. These findings suggest that an amelioration of mitochondrial quality, possibly mediated by mitochondria-derived vesicle recycling, may be among the mechanisms supporting improvements in physical performance and endothelial function during the resolution of long COVID. Full article
(This article belongs to the Special Issue Advances in Anti-Aging Treatment Development, 2nd Edition)
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12 pages, 623 KiB  
Article
PPARγ Expression in Human Spermatozoa and Its Relationship with Seminal F2-Isoprostanes and Resolvin D1 in the Presence of Varicocele and Urogenital Infections
by Giulia Collodel, Elena Moretti, Caterina Marcucci, Laura Liguori, Daniela Marchini, Roberta Corsaro, Gabriele Centini and Cinzia Signorini
Biology 2025, 14(2), 137; https://doi.org/10.3390/biology14020137 - 28 Jan 2025
Viewed by 544
Abstract
Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) is a regulating agent in antioxidant response also involved in controlling inflammation. The impact of varicocele and urogenital infections on sperm PPARγ expression was studied. The PPARγ gene expression was investigated in spermatozoa of 26 normozoospermic men grouped [...] Read more.
Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) is a regulating agent in antioxidant response also involved in controlling inflammation. The impact of varicocele and urogenital infections on sperm PPARγ expression was studied. The PPARγ gene expression was investigated in spermatozoa of 26 normozoospermic men grouped according to their clinical conditions: normal semen parameters (N), normal semen parameters and varicocele (N + V), and normal semen parameters and urogenital infections (N + UI). Sperm PPARγ expression was correlated with F2-isoprostanes (F2-IsoPs), as markers of lipid peroxidation, and Resolvin D1 (RvD1), a pro-resolving mediator in inflammation. Sperm PPARγ expression was evaluated through comparative real-time PCR, and F2-IsoPs and RvD1 were quantified in the seminal plasma via GC/NCI-MS/MS and immunoassay, respectively. PPARγ expression correlates positively with sperm morphology and vitality and negatively with F2-IsoPs and RvD1. Sperm morphology positively correlates with vitality and negatively with F2-IsoP and RvD1 levels. Despite the normozoospermia in the three examined groups, sperm morphology and PPARγ expression were significantly reduced in N + V and N + UI groups compared to the N group. Additionally, F2-IsoP and RvD1 levels were elevated in N + V and N + UI patients. These data suggest that PPARγ expression is compromised by inflammation and lipoperoxidation, providing new insights to further explore new possibilities of targeted treatment of male infertility. Full article
(This article belongs to the Section Developmental and Reproductive Biology)
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13 pages, 1975 KiB  
Article
Novel Automated Chemiluminescent Immunoassay for the Detection of Autoantibodies Against Aquaporin-4 in Neuromyelitis Optica Spectrum Disorders
by Nozomi Yamazaki, Toshiyuki Takahashi, Tatsuro Misu and Yukihiro Nishikawa
Diagnostics 2025, 15(3), 298; https://doi.org/10.3390/diagnostics15030298 - 27 Jan 2025
Viewed by 548
Abstract
Background/Objectives: Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune-related neurological disease that primarily affects the optic nerve and spinal cord. According to current international consensus guidelines for NMOSD, confirming the presence of aquaporin-4 immunoglobulin G antibody (AQP4-IgG) is one of the most [...] Read more.
Background/Objectives: Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune-related neurological disease that primarily affects the optic nerve and spinal cord. According to current international consensus guidelines for NMOSD, confirming the presence of aquaporin-4 immunoglobulin G antibody (AQP4-IgG) is one of the most important diagnostic criteria because AQP4-IgG is a significant diagnostic biomarker of NMOSD. Several assays are currently available for detecting AQP4-IgG, including cell-based assays (CBAs) and enzyme-linked immunosorbent assays (ELISAs). However, each assay has certain disadvantages, including insufficient sensitivity and specificity, the need for sophisticated techniques, and semi-quantitative results. Methods: We developed a fully automated chemiluminescent enzyme immunoassay (CLEIA) to detect AQP4-IgG (AQP4-CLEIA). This assay utilizes the recombinant antigen purified from the newly generated AQP4-M23 stably expressing Chinese hamster ovary cell line and an anti-AQP4 monoclonal antibody as a calibrator. Results: In analytical performance studies, the assay demonstrates good precision and linearity over the entire measurement range. Moreover, this assay showed no high-dose hook effect and interference from endogenous substances. In clinical validation studies, patients with AQP4-IgG positive NMOSD, multiple sclerosis, or myelin oligodendrocyte glycoprotein antibody-associated disorder and healthy individuals were tested. A cutoff value of 10.0 U/mL was determined by receiver operating characteristic curves based on the results of a microscopic live CBA. The sensitivity and specificity for AQP4-IgG-positive NMOSD were 97.0% and 100.0%, respectively, at the cutoff value. Conclusions: The results suggest that AQP4-CLEIA is a convenient automated method for measuring AQP4-IgG titers in hospitals and clinical laboratories, offering an effective alternative to the gold-standard CBA. Full article
(This article belongs to the Special Issue Diagnostic Challenges in Neuroimmunology)
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15 pages, 1863 KiB  
Article
Blood-Based Lateral-Flow Immunoassays Dipstick Test for Damaged Mitochondrial Electron Transport Chain in Pyruvate Treated Rats with Combined Blast Exposure and Hemorrhagic Shock
by Evans Okonkwo, Biswajit Saha, Geetaram Sahu, Alakesh Bera and Pushpa Sharma
J. Clin. Med. 2025, 14(3), 754; https://doi.org/10.3390/jcm14030754 - 24 Jan 2025
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Abstract
Blast trauma presents a unique challenge due to its complex mechanism of injury, which impacts the brain and other vital organs through overpressure waves and internal bleeding. Severe blood loss leads to an inadequate oxygen supply and insufficient fuel delivery to cells, impairing [...] Read more.
Blast trauma presents a unique challenge due to its complex mechanism of injury, which impacts the brain and other vital organs through overpressure waves and internal bleeding. Severe blood loss leads to an inadequate oxygen supply and insufficient fuel delivery to cells, impairing ATP production by mitochondria—essential for cell survival. While clinical symptoms of metabolic disruption are evident soon after injury, the molecular, cellular, and systemic damage persists for days to years post-injury. Current challenges in treating traumatic brain injury (TBI) stem from (1) the lack of early blood-based biomarkers for detecting metabolic failure and mitochondrial damage and (2) the limited success of mitochondrial-targeted therapeutic strategies. Objectives: To identify blood-based mitochondrial biomarkers for evaluating the severity of brain injuries and to investigate therapeutic strategies targeting mitochondria. Methods: A preclinical rat model subjected to blast exposure, with or without hemorrhagic shock (HS), followed by resuscitation was utilized. Blood samples were obtained at baseline (T0), post-injury (T60), and at the conclusion of the experiment (T180), and analyzed using a validated dipstick assay to measure mitochondrial enzyme activity. Results: Blast and HS injuries led to a significant decrease in the activity of mitochondrial enzymes, including complex I, complex IV, and the pyruvate dehydrogenase complex (PDH), compared to baseline (p < 0.05). Concurrently, blood lactate concentrations were significantly elevated (p < 0.001). An inverse correlation was observed between mitochondrial enzyme dysfunction and blood lactate levels (p < 0.05). Treatment with sodium pyruvate post-injury restored complex I, complex IV, and PDH activity to near-baseline levels, corrected hyperlactatemia, and reduced reactive oxygen species (ROS) production by mitochondria. Conclusions: Serial monitoring of blood mitochondrial enzyme activity, such as complex I, complex IV, and PDH, may serve as a valuable tool for prognostication and guiding the use of mitochondrial-targeted therapies. Additionally, mitochondrial enzyme assays in blood samples can provide insights into the global redox status, potentially paving the way for novel therapeutic interventions in TBI. Full article
(This article belongs to the Special Issue Traumatic Brain Injury (TBI): Clinical Updates and Perspectives)
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