Enzyme-linked Immunosorbent Assay Research Articles

Study of rotavirus genotypes G and P in one Egyptian center-cross-sectional study

Abstract Background Rotavirus-associated gastroenteritis is a common health problem in children, different variations of rotavirus genotypes differ according to geographic locations and the practice of wide-scale vaccination. Therefore, the present study aimed to detect both the G and P genotypes of rotavirus in children ≤ 5 years old in one center in Egypt as a cross-sectional study, to correlate the genotypes with various demographic and clinical data in infected children and to evaluate the common mixed genotypes G and P in infected children. Method The cross-sectional study included children with acute gastroenteritis ≤ 5 years old from January 2023 till March 2024 recruited from Mansoura University Children’s Hospital, Egypt based upon laboratory diagnosis by exclusion of bacterial and protozoa pathogens. The stool samples were obtained from each child and subjected to detection of rotavirus antigen by enzyme-linked immunosorbent assay (ELISA) followed by genotypes identification of G and P genotypes by nested polymerase chain reaction (PCR). Result A nested PCR study for rotavirus genotypes revealed that G1 was the most common genotype (24.7%) followed by G2 (21.1%), G3 (20%), G9 (20%), and G4 (14.1%). The genotyping of the P genotype revealed that P9 was the commonest genotype (24.7%), followed by P4 (21.2%), P10 (20%), P8 (17.6%) and P6 (16.5%). The commonest combined genotypes of G and P were G1P4 (85.7%), G3P8(88.2%), followed by G2P6 (77.8%) and G9P9(76.5%) and G4P9 (66.7%) followed by G4P10 (33.3%), G9P10(23.5%), G2P10(22.2%), G1P10 (14.3%), G3P10(11.8%). The distribution was significant (P = 0.001). The positive rotavirus antigen was more frequently detected in females (55.3%) than males (44.7%, Odd ratio 0.2, 95% CI 0.22–0.71, P = 0.001). There was a significant association between the summer season and positive rotavirus antigen (P = 0.001) and rural residence of the patients (Odd ratio 6,9 95%CI 3,5-13.5, P = 0.001). The significant associated clinical sign with positive rotavirus antigen was fever (Odd ratio 3,3, 95%CI 1,8-6.05, P = 0.001). The genotypes G and P were significantly associated with positive rotavirus antigen as all cases positive by antigen had been detected by nested PCR with the commonest genotypes G4 (24.7%, P = 0.001) and genotype P9 (24.7%, P = 0.001). Conclusion The present study highlights the common genotypes of rotavirus at one center in Egypt, G1, G2, and G3 were the commonest G genotypes. As regard genotype P the commonest genotypes were P9, P4, and P10. The commonest combined genotypes were G1P4, G3P8, G2P6. There was no effect of the practice of rotavirus vaccination at limited rates at private health sections as the rotavirus is still a major pathogen of acute gastroenteritis in children. There is a need for the inclusion of rotavirus vaccination in the national program of children vaccination in Egypt.

Assessment of Serum Adiponectin and Leptin in Chronic Preiodontitis Over Healthy Individuals

ABSTRACT Objective: The objective of the research was to ascertain whether serum levels of the biochemical markers adiponectin and leptin are related to clinical periodontal characteristics. Materials and Method: For the study, 20 people with chronic periodontitis (Group A) and 20 healthy people (Group B) were enrolled. Assessments were conducted on periodontal markers, including bleeding on probing (POP), gingival index (GI), plaque index (PI), probing pocket depth (PPD), and clinical attachment level (CAL). An enzyme-linked immunosorbent assay (ELISA) test was used to assess the levels of adiponectin and leptin in serum samples. Result: There was significant elevation in average level of leptin in chronic periodontitis in association to that in healthy group. On the other hand, there was considerable decrease in serum adiponectin in group A in association to healthy group. The ratio of leptin\adiponectin was appreciably greater amongst periodontitis group compared to healthy group. Conclusion: The current investigation leads us to the conclusion that the pathophysiology of periodontitis is significantly influenced by serum levels of adiponectin and leptin.

Innovative Approaches for Sustainable Greenhouse Pest Management: Integrating Advanced Diagnostics and Preventive Strategies for Optimal Crop Health

Sustainable greenhouse pest management is critical for achieving good crop yields while reducing environmental impact. This paper investigates novel ways that incorporate improved diagnostic tools and preventive tactics into the framework of Integrated Pest Management (IPM) in greenhouse ecosystem. Understanding pest life cycles and utilizing cutting-edge tools such as DNA barcoding and enzyme-linked immunosorbent assays (ELISA) might help greenhouse managers achieve precise pest diagnosis and early responses. Preventive measures, such as tight cleanliness protocols, quarantine techniques, and the use of physical barriers, are critical in lowering pest incidence. Furthermore, biological controls, cultural methods such as crop rotation and intercropping, and the selective use of insecticides and biopesticides collectively help to ensure long-term pest management. This complete approach not only reduces insect damage but also creates a healthier, more resilient agricultural ecology. Furthermore, traditional diagnostic and preventive measures were also discussed in the article. The and recommendations are validated by recent studies and data, demonstrating the success of these integrated tactics in modern greenhouse operations.

Seroprevalence and associated risk factors of peste des petits ruminants in sheep and goats in three districts of the Central Oromia Region, Ethiopia

BackgroundPeste des petits ruminants (PPR) is a viral disease that affects domestic and wild small ruminants and camels in Africa, the Middle East, and Asia. Following the successful eradication of rinderpest, the World Organization for Animal Health (WOAH) and the Food and Agriculture Organization (FAO) have undertaken to eradicate PPR by 2030. Regular surveillance and monitoring of the disease in various regions of Ethiopia are crucial to achieving this goal. This study aimed to estimate the prevalence of PPR, assess community awareness of PPR infection, and examine the associated risk factors of the disease in selected districts of the Central Oromia Region, Ethiopia.MethodThe study collected 384 serum samples from 73 flocks containing 217 sheep and 167 goats using a multi-stage sampling technique. A competitive enzyme-linked immunosorbent assay (c-ELISA) was used to detect antibodies against the PPR virus. Additionally, a pre-tested questionnaire was used to gather information on community awareness and potential risk factors for PPRV infection in the study area.ResultsThe study found that the overall prevalence of PPR in flocks was 71.2% [95% confidence interval (CI): 59.4%−81.2%]. The prevalence of PPR at the animal level was 50% (95% CI: 44.9%−55.1%), with sheep having a prevalence of 54.4% (95% CI: 47.0%−60.6%) and goats having a prevalence of 44.3% (95% CI: 36.6%−52.2%). The study also found that districts, flock size, and agroecology were independent predictors of PPRV seropositivity in sheep, whereas districts, origin, and mixed species were independent predictors of PPRV seropositivity in goats.ConclusionThe study revealed a high prevalence of PPR in sheep and goats in the study area. To prevent the spread of the disease, the study suggests quarantining animals before introducing them to districts, regular PPR vaccination, and isolation and molecular characterization of the PPR virus circulating in the study area.

Multi-omic molecular characterization and diagnostic biomarkers for occult hepatitis B infection and HBsAg-positive hepatitis B infection

BackgroundThe pathological and physiological characteristics between HBsAg-positive HBV infection and occult hepatitis B infection (OBI) are currently unclear. This study aimed to explore the immune microenvironment in the peripheral circulation of OBI patients through integration of proteomic and metabolomic sequencing, and to identify molecular biomarkers for clinical diagnosis of HBsAg-positive HBV and OBI.MethodsThis research involved collection of plasma from 20 patients with OBI (negative for HBsAg but positive for HBV DNA, with HBV DNA levels < 200 IU/mL), 20 patients with HBsAg-positive HBV infection, and 10 healthy individuals. Mass spectrometry-based detection was used to analyze the proteome, while nuclear magnetic resonance spectroscopy was employed to study the metabolomic phenotypes. Differential molecule analysis, pathway enrichment and functional annotation, as well as weighted correlation network analysis (WGCNA), were conducted to uncover the characteristics of HBV-related liver disease. Prognostic biomarkers were identified using machine learning algorithms, and their validity was confirmed in a larger cohort using enzyme linked immunosorbent assay (ELISA).ResultsHBsAg-positive HBV individuals showed higher ALT levels (p=0.010) when compared to OBI patients. The influence of HBV infection on metabolic functions and inflammation was evident through the analysis of distinct metabolic pathways in HBsAg-positive HBV and OBI groups. Tissue tracing demonstrated a connection between Kupffer cells and HBsAg-positive HBV infection, as well as between hepatocytes and OBI. Immune profiling revealed the correlation between CD4 Tem cells, memory B cells and OBI, enabling a rapid response to infection reactivation through cytokine secretion and antibody production. A machine learning-constructed and significantly expressed molecule-based diagnostic model effectively differentiated HBsAg-positive and OBI groups (AUC values > 0.8). ELISA assay confirmed the elevation of FGB and FGG in OBI samples, suggesting their potential as biomarkers for distinguishing OBI from HBsAg-positive infection.ConclusionsThe immune microenvironment and metabolic status of HBsAg-positive HBV patients and OBI patients vary significantly. The machine learning-based diagnostic model described herein displayed impressive classification accuracy, presenting a non-invasive means of differentiating between OBI and HBsAg-positive HBV infections.

Melatonin attenuates cardiac dysfunction and inflammation in dilated cardiomyopathy via M2 macrophage polarization

Melatonin is a neuroendocrine hormone that exerts protective effects on the heart. Increasing evidence suggests that macrophage M2-type polarization improves myocardial regeneration and repair. Therefore, this study investigated whether melatonin ameliorates dilated cardiomyopathy (DCM) by modulating M2-type polarization. DCM mice were established by induction with doxorubicin and then treated with melatonin. Cardiac dysfunction was determined by measuring left ventricular ejection fraction (LVEF) and left ventricular internal dimensions at end-diastole and end-systole. Heart injury and fibrosis were determined by hematoxylin and eosin staining and Sirius Red staining, respectively. Serum concentrations of melatonin, tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) were measured through enzyme-linked immunosorbent assays. M2-type macrophages were analyzed by flow cytometry. Relative mRNA and protein levels were determined by reverse transcription quantitative polymerase chain reaction and western blotting, respectively. Circulating melatonin levels were significantly decreased in DCM mice and were associated with LVEF. Treatment with melatonin markedly ameliorated cardiac dysfunction, improved survival, and alleviated pathological changes and collagen deposition in DCM mice. Furthermore, melatonin-treated DCM mice displayed lower serum and cardiac levels of IL-1β, IL-6, and TNF-α, as well as higher numbers of M2-type macrophages in cardiac tissue, indicating that melatonin treatment could decrease inflammatory responses and facilitate M2 macrophage polarization in DCM mice. Thus, melatonin treatment alleviated cardiac dysfunction and inflammatory responses by promoting M2 macrophage polarization in the DCM mouse model.

Effects of CFTR-ENaC on spinal cord edema after spinal cord injury

Abstract Objective To explore the role of cystic fibrosis transmembrane conduction regulator (CFTR)-Epithelial sodium channel (ENaC) in spinal cord edema after spinal cord injury (SCI) and the related mechanism. Methods Lipopolysaccharide (LPS)-treated M1830 astrocytes were applied as the SCI in vitro model. Immunohistochemistry, real-time PCR, and Western blotting were utilized to detect CFTR and ENaC expression. Enzyme-linked immunosorbent assay was used to measure inflammatory cytokines including TNF-α, IL-1β, IL-6, and IL-18. Transmission electron microscope examined ultrastructure changes, while CFTR-172 or Capsazepine treatment assessed their effects on edema and inflammation. Western blot analysis was employed to evaluate the PI3K, p-PI3K, AKT, and p-AKT signaling pathways in treated cells. Results LPS-treated M1830 cells exhibited increased levels of CFTR and pro-inflammatory cytokines, including TNF-α, IL-1β, IL-6, and IL-18, alongside decreased ENaC expression and suppressed p-PI3K/PI3K and p-AKT/AKT levels. Degeneration of the myelin sheath and axons was observed in LPS-treated M1830, while changes in ultrastructural were recovered after adding CFTR-172 or Capsazepine. The level of CFTR, TNF-α, IL-1β, IL-6, and IL-18 was decreased, while the level of ENaC, p-PI3K/PI3K, and p-AKT/AKT was increased obviously in LPS-treated M1830 with CFTR-172, Capsazepine, or IGF-1. Conclusion Down-regulation of CFTR and up-regulation of ENaC can attenuate inflammation in SCI by activating the PI3K/AKT signaling pathway, highlighting a new therapeutic approach for SCI treatment. These findings address a critical gap in current SCI treatments and suggest a novel intervention strategy targeting ion channel regulation.

Integration of Machine Learning and Experimental Validation to Identify Anoikis-Related Prognostic Signature for Predicting the Breast Cancer Tumor Microenvironment and Treatment Response

Background/Objectives: Anoikis-related genes (ANRGs) are crucial in the invasion and metastasis of breast cancer (BC). The underlying role of ANRGs in the prognosis of breast cancer patients warrants further study. Methods: The anoikis-related prognostic signature (ANRS) was generated using a variety of machine learning methods, and the correlation between the ANRS and the tumor microenvironment (TME), drug sensitivity, and immunotherapy was investigated. Moreover, single-cell analysis and spatial transcriptome studies were conducted to investigate the expression of prognostic ANRGs across various cell types. Finally, the expression of ANRGs was verified by RT-PCR and Western blot analysis (WB), and the expression level of PLK1 in the blood was measured by the enzyme-linked immunosorbent assay (ELISA). Results: The ANRS, consisting of five ANRGs, was established. BC patients within the high-ANRS group exhibited poorer prognoses, characterized by elevated levels of immune suppression and stromal scores. The low-ANRS group had a better response to chemotherapy and immunotherapy. Single-cell analysis and spatial transcriptomics revealed variations in ANRGs across cells. The results of RT-PCR and WB were consistent with the differential expression analyses from databases. NU.1025 and imatinib were identified as potential inhibitors for SPIB and PLK1, respectively. Additionally, findings from ELISA demonstrated increased expression levels of PLK1 in the blood of BC patients. Conclusions: The ANRS can act as an independent prognostic indicator for BC patients, providing significant guidance for the implementation of chemotherapy and immunotherapy in these patients. Additionally, PLK1 has emerged as a potential blood-based diagnostic marker for breast cancer patients.

Isofraxidin alleviated radiation-induced testicular damage via the Nrf2/HO-1-NLRP3/ASC Axis

OBJECTIVE: Radiotherapy is used to treat patients with tumors; however, radiation (IR)-induced testicular injury, which has no effective treatment approved in clinical practice, significantly influences their prognosis and quality of life. The protective effects and underlying mechanisms of action of isofraxidin (IF) against IR-induced testicular injury were investigated. Methods: A mouse testis injury model was established using 5 Gy irradiation. H&E staining, immunofluorescence staining, and enzyme-linked immunosorbent assay were used to measure DNA damage, apoptosis, inflammatory reactions, and oxidative stress in the testes of mice after irradiation. The effectiveness of IF irradiation on testicular injury was evaluated, and the mechanisms of the related oxidative stress and inflammatory response pathways are discussed. Results: IF can improve IR-induced testicular injury by inhibiting the increased levels of DNA damage, apoptosis rate, oxidative stress, and inflammatory factors. The radioprotective effects of IF on testicular injury are mediated by the stimulation of Nrf2/HO-1 or suppression of NLRP3 inflammasome signaling pathways. In addition, crosstalk between the Nrf2/HO-1 and NLRP3 inflammasome signaling pathways was elucidated, in which the inhibition of the NLRP3 inflammasome was mediated by the activation of Nrf2 signaling with IF upon IR exposure. Conclusion: IF can be a potent radioprotective agent to mitigate testicular damage, and may provide a new therapeutic option to alleviate the side effects of radiotherapy in male patients with tumors.

Abstract 4138733: Predictive Value of Serum Vasoactive Intestinal Peptide for Low-Voltage Areas in Atrial Fibrillation

Introduction: Identifying low-voltage areas (LVAs) within left atrium (LA) is crucial for predicting atrial fibrillation (AF) recurrence after pulmonary vein isolation (PVI). Animal studies have shown that the parasympathetic neurotransmitter vasoactive intestinal peptide (VIP) affects atrial electrophysiological remodeling. Hypothesis: Thus, we hypothesized that elevated serum VIP levels might contribute to the development of LVAs and that serum VIP concentrations could serve as a biomarker for LVA presence in AF patients. Aims: The aim of this study is to investigate the relationship between the blood levels of VIP and the presence of LVAs in AF patients with electroanatomical mapping (EAM). Methods: We conducted an observational, prospective cross-sectional study on AF patients undergoing PVI between 2021 and 2023. Blood samples for VIP were collected before atrial septal puncture, and EAM was performed using CARTO 3® before PVI. VIP concentrations were measured using an enzyme-linked immunosorbent assay kit. Patients were divided into two groups based on LVA presence (≥5% or <5%) as determined by EAM. Results: Among the 108 patients, 51 (47%) had LVA. VIP levels were significantly higher in patients with LVA compared to those without (median [interquartile range]: 335.1 [274.1-456.5] pg/mL vs. 247.7 [196.1-294.0] pg/mL, p<0.001). No difference was observed in VIP levels by LVA sites. Multivariate logistic regression identified VIP and sex as independent predictors of LVA (per 50 pg/mL, odds ratio [OR]: 1.44, 95% confidence interval [CI]: 1.13-1.83, p<0.001, OR: 4.67, 95% CI: 1.69-12.89, p=0.002, respectively). Females with high VIP had lower conduction velocity compared to other groups. (low VIP male: 0.9 [0.6-1.0] m/s, high VIP male: 0.9 [0.7-1.1] m/s, low VIP female: 0.8 [0.7-1.0] m/s, high VIP female: 0.7 [0.3-0.8] m/s, p=0.013). Incorporating VIP levels (≥288 pg/mL) into an existing predictive score (age ≥65 years, female, and LA volume index ≥57 mL/m 2 ) significantly improved LVA detection (area under the curve: 0.784 vs. 0.707, p<0.001). Conclusions: Serum VIP levels are higher in patients with LVAs, and VIP is a reliable predictor of LVA presence. Incorporating VIP levels into existing predictive models enhances the power for identifying LVAs in AF patients. Serum VIP measurement could be valuable for detecting LVAs, improving risk stratification, and tailoring strategies in AF management.

Abstract 4134273: Low 5-hydroxytryptamine levels are associated with adverse outcomes in patients with heart failure with preserved ejection fraction

Background: Heart failure with preserved ejection fraction (HFpEF) is a heterogeneous syndrome and its pathophysiology is not fully understood. A monoamine, 5-hydroxytryptamine (5-HT), is involved in diverse biological functions and suggested to play a role in cardiovascular diseases. However, the clinical relevance of 5-HT in HFpEF remains unclear. Aims: This study aimed to elucidate the clinical significance and prognostic value of 5- HT in patients with HFpEF. Methods: We conducted a prospective study involving 240 consecutive hospitalized patients with HFpEF (mean age 72 years, 52% male). We measured circulating blood 5-HT levels using the enzyme-linked immunosorbent assay method. Clinical and outcome data were collected. Results: Correlation analysis revealed that 5-HT levels were negatively correlated with blood B-type natriuretic peptide concentration and tricuspid regurgitation pressure gradient. When patients were stratified into two groups based on the median 5-HT levels (90.5 ng/mL), Kaplan-Meier analysis showed that HFpEF patients with low blood 5-HT levels had lower event-free survival rates from the composite event of cardiac death and worsening heart failure over a median follow-up period of 725 days (Figure). In a multivariable Cox proportional hazard model adjusting for confounding variables, low levels of 5-HT were independently associated with increased risks of the composite of cardiac events (hazard ratio, 3.25; P < 0.01). Conclusion: Low 5-HT levels are associated with adverse outcomes in patients with HFpEF and 5-HT may serve as a useful biomarker for predicting prognosis in such patients.

First detection of alfalfa mosaic virus in Ficus benjamina in Chile

Ficus benjamina, commonly known as ficus or weeping fig, is an ornamental tree from Southeast Asia in the Moraceae family. It is an evergreen species grown as an interior plant worldwide, prized for its glossy green leaves, gray stem, and multiple branches. F. benjamina can be pruned and kept as a small shrub when grown indoors, with most commercially available cultivars in the ornamental industry derived from sports and propagated cuttings (Chen et al., 2010). In October 2022, symptoms indicative of alfalfa mosaic virus (AMV) infection, such as intense bright yellow spots, mottling and chlorosis were observed in leaves of F. benjamina growing in office environments in Santiago, Chile (supplementary Fig.1). Seven symptomatic and three asymptomatic plants were sampled and traced back to the same plant nursery, and only leaf tissue samples from the symptomatic plants tested positive for the presence of the AMV using enzyme-linked immunosorbent assay (ELISA) (Agdia Inc., Elkhart, IN). These results were verified using reverse transcription (RT)-PCR with specific primers AMV-F and AMV-R, targeting a region of the AMV CP gene (Xu and Nie, 2006). AMV was detected in all symptomatic leaf samples, producing 351-bp amplicons in the RT-PCR assay for all samples that were positive to AMV by ELISA. No amplification product was observed when seronegative samples or non-template control was used as templates in the RT-PCR assays. Three RT-PCR amplicons were directly sequenced in both directions. BLAST analysis of these sequences showed 100% nucleotide sequence identity to an AMV isolate previously reported (GenBank Accession No. KX458469), confirming the detection of AMV in F. benjamina. This pathogen causes disease in several species, including ornamentals plant such as Viburnum tinus L. (Peña et al., 2011), peony (Belardi and Rubies, 2003), phlox (Holcomb et al., 2006), and weeds such as Cayratia japonica (Thunb.) Gagnep, Justicia procumbens L. and Veronica persica Poir. (Gao et al., 2020). F. benjamina is among the most popular ornamental indoor plants in Chile, so nurseries should ensure AMV-free plants to restrict disease incidence and prevent the spread of this virus. However, Chile lacks specific regulations for ornamental nurseries, making adherence to pest and disease recommendations voluntary, and only imported plant material requires a sanitary certificate.

Interferon gamma kinetics associated with immunodiagnostic techniques in experimental goats infected with biofilm producer and non-biofilm producer strains of Corynebacterium pseudotuberculosis

Caseous lymphadenitis can be diagnosed using serological tests, mainly the enzyme-linked immunosorbent assay (ELISA). In this work, we evaluated interferon-gamma production in goats infected and not infected with Corynebacterium pseudotuberculosis, correlating it with the ELISA diagnostic test and analyzing the cellular and humoral responses, respectively. Eighteen Canindé goats experimentally infected with biofilm-producing and non-biofilm-producing strains were used. The sensitivity of the antigens was above 80%, with seroconversion observed from 14 days post-infection. A significant band between 64 and 70 kDa was shown in Western blotting. The production of IFN-y shows a tendency towards greater production in goats infected with the biofilm-producing strain stimulated by the TPP antigen of the same strain. The results help better understand the cellular and humoral response and antigenicity of the strains under study, correlated with the production of IFN-y, which plays a vital role in infection.

Assessment of the Concentration of Transforming Growth Factor Beta 1–3 in Degenerated Intervertebral Discs of the Lumbosacral Region of the Spine

The purpose of this study was to evaluate the feasibility of using the expression profile of transforming growth factor beta (TGF-β-1-3) to assess the progression of L/S spine degenerative disease. The study group consisted of 113 lumbosacral (L/S) intervertebral disc (IVD) degenerative disease patients from whom IVDs were collected during a microdiscectomy, whereas the control group consisted of 81 participants from whom IVDs were collected during a forensic autopsy or organ harvesting. Hematoxylin and eosin staining was performed to exclude degenerative changes in the IVDs collected from the control group. The molecular analysis consisted of reverse-transcription real-time quantitative polymerase chain reaction (RT-qPCR), an enzyme-linked immunosorbent assay (ELISA), Western blotting, and an immunohistochemical analysis (IHC). In degenerated IVDs, we noted an overexpression of all TGF-β-1-3 mRNA isoforms with the largest changes observed for TGF-β3 isoforms (fold change (FC) = 19.52 ± 2.87) and the smallest for TGF-β2 (FC = 2.26 ± 0.16). Changes in the transcriptional activity of TGF-β-1-3 were statistically significant (p < 0.05). Significantly higher concentrations of TGF-β1 (2797 ± 132 pg/mL vs. 276 ± 19 pg/mL; p < 0.05), TGF-β2 (1918 ± 176 pg/mL vs. 159 ± 17 pg/mL; p < 0.05), and TGF-β3 (2573 ± 102 pg/mL vs. 152 ± 11 pg/mL) were observed in degenerative IVDs compared with the control samples. Determining the concentration profiles of TGF-β1-3 appears to be a promising monitoring tool for the progression of degenerative disease as well as for evaluating its treatment or developing new treatment strategies with molecular targets.

Assessing the predictive potential of ADAM8 for disease control in chronic rhinosinusitis with nasal polyps

BackgroundA disintegrin and metalloproteinase 8 (ADAM8) has been implicated in eosinophilic inflammation; however, its role in chronic rhinosinusitis with nasal polyps (CRSwNP) remains to be elucidated. This study aimed to investigate the predictive significance of ADAM8 levels in nasal secretions for the endotypes and disease control status of CRSwNP.MethodsA cohort comprising 120 CRSwNP patients and 45 healthy controls (HCs) was assembled, delineating 53 non-eosinophilic CRSwNP (neCRSwNP) and 67 eosinophilic CRSwNP (eCRSwNP) patients. Immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) were utilized to measure ADAM8 levels in nasal mucosal tissues and secretions from all participants. The receiver operating characteristic (ROC) curves and Pearson correlation analysis were employed to assess the predictive capability of ADAM8 levels in predictiving CRSwNP endotypes and disease control status.ResultsADAM8 levels in nasal secretions were elevated in CRSwNP patients compared to HCs, with a more pronounced increase observed in eCRSwNP patients. Elevated ADAM8 concentrations in nasal secretions were positively correlated with peripheral blood eosinophil counts and percentages, tissue eosinophil counts, serum total IgE, Lund–Mackay scores, and Lund-Kennedy scores. Ultimately, 103 CRSwNP patients completed the follow-up protocol, with 72 classified as the controlled group and 31 as the uncontrolled group. Uncontrolled CRSwNP patients exhibited significantly higher ADAM8 levels in nasal secretions compared to the controlled group. The ROC curves indicated that ADAM8 in nasal secretions exhibits robust discriminatory capacity for eCRSwNP and postoperative disease control status.ConclusionADAM8 in nasal secretions emerges as a potential novel biomarker for the prognostication of CRSwNP endotypes and the postoperative disease control status.

Potential of epicatechin as antioxidant and antiaging in UV-induced BJ cells by regulating COL1A1, FGF-2, GPX-1, and MMP-1 gene, protein levels, and apoptosis

Background Oxidative stress caused by exposure to ultraviolet (UV) light on the skin can damage deoxyribonucleic acid (DNA) and cause keratinocytes to undergo apoptosis. Endogenous antioxidants which play a role in trapping free radicals are also unable to overcome excess reactive oxygen species (ROS) in the body due to UV exposure, so exogenous antioxidants are needed. Polyphenolic compounds extracted from natural ingredients such as flavonoids, quercetin, and epicatechin have quite strong antioxidant activity. This is influenced by the chemical structure of these compounds which are rich in hydroxyl groups and aromatic groups. This structure allows the compound to become an electron donor so that it can neutralize free radicals. In vitro research was used to see the potential effectiveness of epicatechin as an antiaging and antioxidant. The study aims to confirm the potential of epicatechin as an antiaging by in vitro assay. Methods The viability test of epicatechin on human skin fibroblast (BJ) cells was carried out using the water-soluble tetrazolium (WST) assay. BJ cells were UV-induced as a cell model of premature aging. Epicatechin 6.25, 12.5, and 25 µg/mL were administered to UV-induced BJ cells. The gene expression of Collagen I Alpha 1 (COL1A1), matrix metalloproteinase-1 (MMP-1), fibroblast growth factor-2 (FGF-2), and glutathione peroxidase-1 (GPX-1) were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Elastin (ELN), hyaluronidase (HAase), cyclooxigenase-2 (COX-2), 8-hydroxydeoxyguanosine (8-OhdG), and melatonin (MT) protein levels were analyzed by enzyme-linked immunosorbent assay (ELISA). The apoptosis of BJ cells was analyzed using flow cytometry. Results Treatment with epicatechin increased relative gene expression including COL1A1 (5.94), FGF-2 (8.34), and GPX-1 (8.09), and also decreased MMP-1 (2.90) relative gene expression compared to the UV-induced BJ cells. Epicatechin also increased levels of ELN (107.7 ng/mg protein) and MT (830 ng/mg protein) levels compared to the UV-induced BJ cells. Epicatechin treatment decreased levels of HAase (505.96 ng/mg protein), COX-2 (33.69 ng/mg protein), and 8-OHdG (97.87 ng/mg protein) compared to the UV-induced BJ cells. Epicatechin also succeeded in maintaining the percentage of live cells and reducing apoptosis, necrotic of UV-induced skin fibroblast cells. Conclusions Epicatechin has the potential to be an antiaging agent by in vitro assay.

Serological and Molecular Investigation of Infectious Laryngotracheitis Virus in Chickens from Robe Town, Southeastern Ethiopia

Infectious laryngotracheitis virus (ILTV) is responsible for avian infectious laryngotracheitis (ILT), a highly contagious acute respiratory disease affecting chickens. However, there is limited information on ILTV and its distribution in Ethiopia, particularly in the southeastern region. The aim of this study was to establish the serological prevalence and molecular evidence in commercial and backyard chickens from Robe town, Southeastern Ethiopia. A cross-sectional study was conducted between December 2021 and June 2022, collecting 240 serum samples from randomly selected chickens belonging to eight kebeles (farms) using systematic random sampling. ILTV-specific antibodies were detected using a commercial indirect enzyme-linked immunosorbent assay (ELISA). From 240 serum samples, 26.7% were positive for ILTV antibodies. Logistic regression analysis identified the type of poultry farm (backyard) and the introduction of chickens from other farms as potential risk factors associated with ILTV exposure. Tracheal tissue and oropharyngeal and tracheal swabs were collected from suspected chickens for isolation and molecular detection. A total of six samples were successfully isolated in embryonated eggs (40%), with four of them verified with a specific PCR. These findings documented the presence of ILTV in the study area, which needs further insight to fully understand the actual spread of ILTV and quantify the damage caused to the poultry sector.

Seroprevalence and silent infection rate during SARS-CoV-2 pandemic among children and adolescents in Western Pomerania: a multicenter, cross-sectional study—the COVIDKID study

Background Limited data on SARS-CoV-2 seroprevalence in rural areas of northern Germany necessitate comprehensive cohort studies. We aimed to evaluate the seroprevalence, silent infection (SI) rates and risk factors for infections among children and adolescents in Western Pomerania from December 2020 to August 2022. Methods In this cross-sectional study, serum or plasma samples from children and adolescents (6 months to 17 years) were collected during routine blood draw. SARS-CoV-2 specific antibodies (S1 and nucleocapsid) and their neutralizing capacity were analyzed using commercially available enzyme-linked immunosorbent and neutralization assays. Socio-demographic data and information about SARS-CoV-2 infection or vaccination were obtained. Multivariable logistic regression was used to identify independent risk factors for SARS-CoV-2 infections and SI. Results A total of 1,131 blood samples were included into the study. Overall, SARS-CoV-2 seroprevalence was 25.1%, strongly influenced by the pandemic course, predominant virus variants, age and approval of vaccination. SI rate was 5.4% (95%-CI [3.7%–6.8%]) among unvaccinated and undiagnosed children over the entire study period with highest rates among adolescents. Main risk factor despite the time at risk for silent infections was an infected household member (Odds ratio = 9.88, 95%-CI [4.23–22.9], p < 0.001). Factors associated with overall infections (known and silent) also include the infection of a household member (Odds ratio = 17.8, 95%-CI [10.7–29.6], p < 0.001). Conclusions We believe that the introduction of governmental measures and systematic test strategies in schools strongly impacted on the SI rate, as we suspect that asymptomatic cases have already been identified, resulting in surprisingly low SI identified in our study.

Does salivary cortisol serve as a potential biomarker for temporomandibular disorders in adults?

BackgroundThe etiology of temporomandibular disorders (TMD) is multifactorial, involving a complex interplay of psychological and physiological factors. While salivary biomarkers, particularly cortisol, play an important role in TMD pathophysiology, evidence in the literature is still scarce and inconsistent. Hence, this study aims to evaluate the applicability of salivary cortisol as a potential biomarker for TMD in adults.MethodsThe Diagnostic Criteria for Temporomandibular Disorders (DC/TMD) were used to accurately diagnose TMD patients. The study included adults, both male and female, aged 18–40 in the TMD group (n = 66) and non TMD participants (n = 66) matched for age and gender. Salivary samples were collected from participants at two time points: early and late morning. Cortisol levels in the samples were quantified using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using a one-way ANOVA to evaluate the correlations between cortisol levels and the study variables. Tukey’s post-hoc tests were applied to adjust for multiple comparisons.ResultsSalivary cortisol levels were significantly higher in the TMD group than in the corresponding controls (p = 0.034). In the TMD group, the mean cortisol levels early in the morning were 29.95 ± 75.05 (0-398.64), while the late morning levels were recorded as 4.87 ± 3.96 (0-17.13). In the control group, the mean cortisol levels early in the morning were 10.98 ± 16.83 (2.16–92.90), and late morning mean cortisol levels were 6.15 ± 6.13 (0-20.42). This indicates that the early morning levels of cortisol are higher in TMD patients (p = 0.046). In the subgroup analysis of the TMD, the mean salivary cortisol levels recorded were highest at 82.49 ± 124.34 (8.32-398.64) in patients having disc displacement without reduction with limited mouth opening. Furthermore, the mean salivary cortisol levels in the early morning were statistically higher (84.83 ± 132.80) in males compared to females (9.36 ± 9.01) (p = 0.008) with TMD.ConclusionThe result of this study suggests that salivary cortisol could be a potential biomarker for a specific TMD subtype (disc displacement without reduction with limited mouth opening). However, further studies are needed to better understand the role of cortisol biomarker in the underlying pathogenesis of TMD.

Analysis of associations of personality traits and stress with the level of serum cortisol in Sakha (Yakut) men

The purpose of this study is to test the hypothesis that personality traits and stressful situations experienced in childhood could be associated with the level of serum cortisol. The sample included 121 healthy adult men of Yakut (Sakha) ethnicity aged 18 to 27 years. To assess personality traits, the TCI temperament and character questionnaire by R. Cloninger was used. Serum cortisol levels were assessed by enzyme-linked immunosorbent assay (ELISA). It was found that such a temperament trait as “reward dependence” is associated with higher level of cortisol in the blood (p = 0.04). Experienced stressful situations are associated with the character trait “self-transcendence” (p = 0.049) but do not significantly affect cortisol levels. In individuals with high levels of stress, significant correlations were found between the “novelty seeking” (r = 0.33) and “self-directedness” (r = 0.36) with serum cortisol levels, which may reflect the prolonged effect of stress on increasing the sensitivity of the adrenal cortex. The results indicate a possible connection between one of the human temperament traits “reward dependence” and higher levels of cortisol in the blood. A high level of experienced stress situations reduces scores on the character trait “self-transcendence.”