Search Results (101)

Celiac disease (CD), a human leukocyte antigen-associated disorder, is caused by gluten sensitivity and is characterized by mucosal alterations in the small intestine. Currently, its diagnosis involves the determination of serological markers. The traditional method for clinically determining these markers is the enzyme-linked immunosorbent assay. However, immunosensors offer sensitivity and facilitate the development of miniaturized and portable analytical systems. This work focuses on developing an amperometric immunosensor for the quantification of IgA antibodies against tissue transglutaminase (IgA anti-TGA) in human serum samples, providing information on a critical biomarker for CD diagnosis. The electrochemical device was designed on a polyimide substrate using a novel solid ink of wax and carbon nanofibers (CNFs). The working electrode microzone was defined by incorporating aminofunctionalized TiO₂ nanoparticles (TiO₂NPs). The interactions and morphology of CNFs/wax and TiO₂NPs/CNFs/wax electrodes were assessed through different characterization techniques. Furthermore, the device was electrochemically characterized, demonstrating that the incorporation of CNFs into the wax matrix significantly enhanced its conductivity and increased the active surface area of the electrode, while TiO₂NPs contributed to the immunoreaction area. The developed device exhibited remarkable sensitivity, selectivity, and reproducibility. These results indicate that the fabricated device is a robust and reliable tool for the precise serological diagnosis of CD. Full article

A large cohort of Romanian children suspected of celiac disease (CD) received comprehensive evaluation through this study regarding serological, genetic, and biochemical markers. This study investigated the relationships between anti-tissue transglutaminase (anti-tTG), anti-endomysium antibodies (EMAs), anti-gliadin deamidated (DGP) antibodies, and HLA genotyping. A strong association was observed between high anti-tTG IgA titers (>100 U/mL) and EMA IgA positivity, with a 95% concordance rate. Furthermore, anti-tTG IgA positive correlated with a significant prevalence of DGP antibodies, suggesting the complementary diagnostic role of DGP antibodies in equivocal cases. Genetic testing for HLA-DQ2/DQ8 alleles validated their association with celiac disease susceptibility, with 50% of the studied patients exhibiting these markers. The research reveals that vitamin D insufficiency affects a large number of children with anti-tTG antibodies, thus requiring both screening and supplementation practices. Furthermore, associations with other autoimmune conditions were explored, including thyroid and diabetes-related autoantibodies. This research demonstrates why CD diagnosis and management require a complete approach that combines serological tests with genetic evaluation and prompt intervention for related health conditions. Full article

Background/Objectives: Coeliac disease is an immune-mediated disorder of the gastrointestinal tract that may result in significant nutritional deficiencies. Effective management requires strict, lifelong adherence to a gluten-free diet. Both underdiagnosis and unnecessary dietary restrictions can adversely affect patients’ health and quality of life. To assess adherence to the current recommendations for the laboratory diagnosis of coeliac disease and promote evidence-based practices while reducing inter-laboratory variability, the Spanish Group on Autoimmunity of the Spanish Society of Immunology conducted a nationwide survey. Methods: A thirty-item survey was distributed to fifty autoimmune laboratories across Spain. Data were collected through a structured Excel-based questionnaire comprising multiple-choice items, which was distributed via email to the participating laboratories. It explored practices related to the diagnosis of coeliac disease in the general population and among at-risk groups as well as approaches to patient follow-up and demand management. Results: Thirty-five laboratories completed the electronic questionnaire. For the serological screening of coeliac disease, all the respondents reported using IgA anti-tissue transglutaminase (tTG-IgA) antibody testing together with total IgA measurement to assess IgA competence. However, consistent use of anti-endomysial antibody testing and HLA genotyping and adherence to pre-analytical recommendations for accurate interpretation of results were not uniform across centres. Conclusions: At the time these data were collected (the third trimester of 2021), the 2020 ESPGHAN guidelines for the diagnosis of coeliac disease in the paediatric population had not yet been fully implemented in most of the laboratories surveyed. For diagnosing adults, most laboratories adhered to local and European guidelines. Full article

Gelsemine, a naturally occurring indole alkaloid derived from plants of the Gelsemium species of the Gelsemiaceae family, has been extensively investigated for its neuroprotective and anti-inflammatory properties. Recent studies have demonstrated that gelsemine exerts neuroprotective effects against beta-amyloid (Aβ) oligomers, a key neurotoxic peptide implicated in the pathogenesis of Alzheimer’s disease (AD). However, despite these beneficial effects, the precise molecular targets underlying gelsemine’s neuroprotective actions in AD remain unidentified. Here, we employed a combination of bioinformatic, biochemical, and functional assays in neuronal models to investigate the mechanism of gelsemine’s action in AD cellular models. Our findings indicate that gelsemine inhibits the activity of transglutaminase 2 (TG2), an enzyme involved in protein cross-linking with emerging roles in Aβ aggregation and neurotoxicity. Molecular modeling and biochemical analyses reveal that gelsemine interacts with the TG2 catalytic site, leading to its inhibition. Furthermore, gelsemine modulates the TG2-mediated Aβ aggregation process, thereby attenuating Aβ-induced neurotoxicity and preserving neuronal function. These findings establish TG2 as a previously unrecognized molecular target of gelsemine and underscore the potential of Gelsemium-derived alkaloids as neuroprotective agents. The modulation of TG2 activity by natural alkaloids may provide a novel therapeutic approach for mitigating Aβ toxicity and preserving neuronal function in AD. Full article

In this study, chitooligosaccharides (COS) were introduced into black soybean protein (BSP) using transglutaminase (TG) as a biocatalyst. The film-processing properties and physiological activities of the enzymatically glycosylated black soybean protein (EGBSP) were studied. The results showed that glycosylation decreased the surface hydrophobicity, absolute value of the zeta potential, its minimum solubility, and film permeability of BSP by 69.86%, 6.04%, 36.68%, and 14.91%, respectively, while increasing the tensile strength and elongation at break of its protein film by 56.57% and 172.68%, respectively. The gel time was shortened, and the acid-induced gel properties of EGBSP were similar to those of BSP. The anticancer effect of EGBSP was evaluated by the tumor inhibition rate, flow cytometry, and morphology observation of an ascites tumor in H₂₂ tumor-bearing mice. The immune organs (spleen, thymus), immune cells (lymphocytes, NK cells), and immune factors (IL-2, IL-12) of H₂₂ tumor-bearing mice were detected to evaluate the immunomodulatory effects of EGBSP. The results showed that medium and high doses of BSP had positive effects on immune enhancement and anti-cancer activity of H₂₂ tumor-bearing mice, while almost all doses of EGBSP showed significant effects. These results indicated that glycosylation significantly improved the anti-cancer effect and immunomodulatory activity of H₂₂ tumor-bearing mice while prolonging their overall survival. In conclusion, the glycosylation method using microbial transglutaminase to catalyze the introduction of chitooligosaccharides into black bean protein can improve the film-processing properties and biological activities of BSP more effectively than the enzyme crosslinking method. Full article

A novel electrochemical detection method utilizing a cost-effective hybrid-modified electrode has been established. A glassy carbon (GC) modified electrode was tested for its ability to measure electrochemical tTG antibody levels, which are essential for diagnosing and monitoring Celiac disease (CD). Tissue transglutaminase protein biomolecules are immobilized on a quantum dots-polypyrrole nanocomposite in the improved electrode. Initial, quantum dots (QDs) were obtained from Bombyx mori silk fibroin and embedded in polypyrrole film. Using carbodiimide coupling, a polyamidoamine (PAMAM) dendrimer was linked with GQDs-polypyrrole film to improve sensor sensitivity. The tissue transglutaminase (tTG) antigen was cross-linked onto PAMAM using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC)-N-hydroxy succinimide (NHS) chemistry to develop a nanoprobe that can detect human serum anti-tTG antibodies. The physicochemical characteristics of the synthesized nanocomposite were examined by FTIR, UV-visible, FE-SEM, EDX, and electrochemical studies. The novel electrode measures anti-tissue antibody levels in real time using human blood serum samples. The modified electrode has great repeatability and an 8.7 U/mL detection limit. Serum samples from healthy people and CD patients were compared to standard ELISA kit assays. SPSS and Excel were used for statistical analysis. The improved electrode and detection system can identify anti-tissue antibodies up to 80 U/mL. Full article

Background: Celiac disease (CD) is an autoimmune disease that results from the interaction of genetic, immune, and environmental factors. According to the 2020 European Society for Pediatric Gastroenterology Hepatology and Nutrition (ESPGHAN) guidelines, an elimination diet (i.e., excluding products that may contain gluten) is the basic method of treating celiac disease. Following a gluten-free diet is extremely problematic, and patients often make unconscious deviations from the diet. According to the current Oslo definitions for celiac disease, depending on the clinical picture and adequate tests, several forms of celiac disease have been identified: typical, atypical, asymptomatic, potential, and refractory. Objective: The aim of the study was to assess the frequency of conscious diet mistakes and unconscious deviations from a gluten-free diet in a group of patients with long-standing celiac disease and their impact on the frequency of typical and atypical symptoms. Methods: The study included 57 people diagnosed with celiac disease between 1980 and 2010. After verifying the history of the disease according to the ESPGHAN guidelines from 2020, we excluded 19 patients who had Marsh grade 1 at the time of diagnosis or those without HLA DQ2 or DQ8 haplotypes detected. After verification, the study included 38 patients, 30 women and 8 men, with a verified diagnosis of typical celiac disease. The effectiveness of the gluten-free diet was assessed in all participants. Blood was collected to determine IgA anti-tissue transglutaminase II antibodies (anti-tTG) and IgG antibodies against deamidated gliadin peptides by ELISA. All survey participants provided data concerning current gastrointestinal and systemic symptoms, bowel habits, comorbidities, dietary habits, physical activity, and socioeconomic conditions. Results: A total of 25 patients (65.78%) declared strict adherence to the gluten-free diet. However, in this group, seven (18.4%) patients had significantly increased levels of anti-tTG antibodies (mean 82.3 RU/mL ± 78.9 SD at N < 20 RU/mL). Among the patients who consciously made dietary mistakes, six (46.2%) demonstrated increased levels of anti-tTG antibodies. The analysis did not reveal any difference between the frequency of intestinal and extraintestinal symptoms in patients making dietary mistakes and following the gluten-free diet. Conclusions: More than half of celiac patients unconsciously or consciously make dietary mistakes, which indicates an urgent need to increase their general knowledge of CD and the appropriate diet. Regardless of whether the gluten-free diet is followed, both typical and atypical symptoms of the disease have been observed among celiac patients. Full article

Background/Objectives: An increasing number of studies have reported liver involvement in both children and adults with celiac disease (CD). This often manifests as isolated hypertransaminasemia or hepatic steatosis (HS). The aim of this study was to define the prevalence of hypertransaminasemia and HS in a pediatric population with CD before starting a gluten-free diet (GFD) and to analyze how the introduction of a GFD could modify this condition. We also conducted a state-of-the-art literature review of the association between hypertransaminasemia, metabolic dysfunction-associated steatotic liver disease (MASLD) and CD. Methods: We retrospectively reviewed the clinical charts of pediatric CD patients diagnosed in three different pediatric units of Sicily, analyzing clinical, laboratory, ultrasound, and histology data before and 12 months after the introduction of a GFD. Results: A total of 160 patients (65.0% females, median age 6.4 (0.8–13.2) years) were included; hypertransaminasemia and HS prevalences at diagnosis were 8.1% and 6.1%, respectively. Subjects with hypertransaminasemia were younger (p = 0.01) than those without and had higher frequencies of HS (p = 0.034) and anti-tissue transglutaminase (tTg) immunoglobulin (Ig)G positivity (p = 0.046). Subjects with HS were younger (p = 0.0001) and had a higher frequency of hypertransaminasemia (p = 0.029) compared to non-steatotic ones. After 12 months of a GFD, hypertransaminasemia and HS persisted in 53.8% and 50.0% of patients, respectively. Conclusions: The prevalences of hypertransaminasemia and HS in Sicilian pediatric CD patients seem to be lower than those reported in other geographical areas. A GFD can reverse the trend of liver involvement, although periods of longer than 12 months may be necessary. However, a GFD has been associated with an increased prevalence of HS, and so regular follow-up involving a nutritionist should be recommended to guide physicians in patient management. Full article

This review explores strategies for mitigating food allergies by treating foods with biopolymers, bioactive compounds, and food-grade enzymes. Biopolymers like chitosan, alginate, and pectin show potential in reducing the allergenic properties of food. Polyphenols such as quercetin, resveratrol, curcumin, and epigallocatechin gallate demonstrate promise as anti-inflammatory molecules that can lessen the symptoms and severity of allergic reactions. Enzymes, including proteases such as pepsin, papain, and bromelain, and transferases like transglutaminase, offer the potential to reduce the allergenic potency of proteins by various mechanisms, though more research is needed for the optimization and assessment of the safety and palatability of treated foods. Overall, this review offers insights into potential strategies to alleviate allergic reactions by reducing the allergenic properties of food proteins. Full article

Background/Objectives: This study aimed to assess the nutritional status of children and adolescents with celiac disease (CD) in Kuwait and investigate the nutritional deficiencies and sociodemographic factors associated with growth stunting in this population. Methods: This case–control study included 77 CD patients aged 3–18 years diagnosed with CD using IgA anti-tissue transglutaminase and duodenal biopsy and 33 healthy controls. Nutritional status was evaluated based on demographic and clinical characteristics, anthropometric measurements, and biochemical parameters. Univariate and multivariate logistic regression models were used to determine the association between CD and growth stunting. Results: Approximately one-third (31%) of children with CD had stunted growth, 20.8% had a low body mass index for their age, and 5.2% had both growth stunting and wasting. Children with CD had higher odds of iron-deficiency anemia, vitamin D deficiency, anemia, and lower socioeconomic status. They were also younger and had decreased serum levels of vitamin D compared to the controls. These factors were all significantly associated with an increased risk of CD, collectively explaining over 50% of the risk. For growth stunting, lower education status among mothers, family income, and serum ferritin were identified as risk factors. Conclusions: A significant proportion of children and adolescents with CD had malnutrition, overt deficiencies, and impaired growth despite coherence with a gluten-free diet. Recommendation: Routine monitoring and targeted nutritional interventions are recommended for children and adolescents with CD to address malnutrition and growth stunting. Addressing socioeconomic disparities and enhancing maternal education may also help mitigate the risk factors. Full article

Fish retinal ganglion cells (RGCs) can regenerate after optic nerve lesions (ONLs). We previously reported that heat shock factor 1 (HSF1) and Yamanaka factors increased in the zebrafish retina 0.5–24 h after ONLs, and they led to cell survival and the transformation of neuro-stem cells. We also showed that retinoic acid (RA) signaling and transglutaminase 2 (TG2) were activated in the fish retina, performing neurite outgrowth 5–30 days after ONLs. In this study, we found that RA signaling and TG2 increased within 0.5 h in the zebrafish retina after ONLs. We examined their interaction with the TG2-specific morpholino and inhibitor due to the significantly close initiation time of TG2 and HSF1. The inhibition of TG2 led to the complete suppression of HSF1 expression. Furthermore, the results of a ChIP assay with an anti-TG2 antibody evidenced significant anti-TG2 immunoprecipitation of HSF1 genome DNA after ONLs. The inhibition of TG2 also suppressed Yamanaka factors’ gene expression. This rapid increase in TG2 expression occurred 30 min after the ONLs, and RA signaling occurred 15 min before this change. The present study demonstrates that TG2 regulates Yamanaka factors via HSF1 signals in the acute phase of fish optic nerve regeneration. Full article

The presenting pattern of celiac disease (CD) at diagnosis in children has changed over time, with a reduction of malabsorption-related phenotypes and an increase in regular or even excessive growth patterns. We retrospectively reviewed the body mass index (BMI) distribution of all patients with a new diagnosis of CD made in a Pediatric Gastroenterology Outpatient Clinic in 1990–2011, compared to those diagnosed in 2012–2022, according to their clinical and serological characteristics. The 1990–2011 and 2012–2022 cohorts included 250 (M:F = 90:160, mean age 7.3 ± 6.1 years) and 243 children (M:F = 81:162, mean age 7.1 ± 3.7 years, NS), respectively. The prevalence of underweight (UW) was higher in the 1990–2011 cohort (61/250, 24.4% in 1990–2011 vs. 31/243, 12.7% in 2012–2022, p = 0.0001), whereas that of overweight (OW) and obese (OB) subjects was significantly higher in 2012–2022 (10/250, 4% in 1990–2011 vs. 24/243, 9.9% in 2012–2022, p = 0.012, and 1/250, 0.4% in 1990–2011 vs. 8/243, 3.3% in 2012–2022, p = 0.018, respectively). In both cohorts, gastrointestinal symptoms were more frequent in OW/OB than in UW children (6/11, 54.5% vs. 5/61, 8.2% in 1990–2011, p < 0.0001, and 24/32, 75% vs. 10/31, 32.3%, p < 0.0001 in 2012–2022), and the extent of anti-transglutaminase antibody increase was similar in OW/OB and UW subjects. The prevalence of children with a normal or even high BMI at CD diagnosis has increased in the past three decades; therefore, CD should be suspected regardless of BMI status. Full article

Background: Celiac disease (CD) is an immune-mediated disease characterized by disruptions of the small intestine. Factors such as viral and bacterial infections can trigger CD. Recently, the reactivation of Human Endogenous Retroviruses (HERVs) has also been implicated, but little is known about their specific role in patients with celiac disease. Methods: The purpose of this study is to explore the humoral immune response mounted against epitopes derived from the envelope portion of three families of HERVs (HERV-K, HERV-H, and HERV-W) in CD patients. Reactivity against the HERV-K, HERV-H, and HERV-W env-su peptides was tested by indirect ELISAs in plasma of 40 patients with celiac disease and 41 age-matched healthy subjects (HCs). Results: HERV-K, HERV-H, and HERV-W env-su peptides triggered different antibody responses in CD patients compared to HCs, with a stronger reactivity (p = 0.0001). Conclusions: Present results show, for the first time, that epitopes of HERV-K, HERV-H, and HERV-W are more recognized in patients with CD. Taking into consideration their proinflammatory and autoimmune features, this might suggest that HERVs may contribute to the development of CD or its exacerbation in genetically predisposed subjects. Finally, to elucidate the interplay between gut inflammation and HERVs during the inflammatory process, further studies are required. Those investigations should focus on the expression levels of HERVs and their relationship with the immune response, specifically examining anti-transglutaminase 2 (TG2) antibody levels under both gluten-free and gluten-containing dietary conditions. Full article

Glioblastoma (GBM) is one of the most aggressive cancers, characterized by a decrease in antioxidant levels. Evidence has demonstrated that ferulic acid (FA), a natural antioxidant particularly abundant in vegetables and fruits, could be a promising candidate for GBM treatment. Since FA shows a high instability that compromises its therapeutic application, it has been encapsulated into Nanostructured Lipid Carriers (NLCs) to improve its bioavailability in the brain. It has been demonstrated that tissue transglutaminase (TG2) is a multi-functional protein implicated in many physiological and pathological processes, including cancer. TG2 is also involved in GBM correlated with metastasis formation and drug resistance. Therefore, the evaluation of TG2 expression levels and its cellular localization are important to assess the anti-cancer effect of FA against GBM cancer. Our results have demonstrated that treatment with free FA and FA-NLCs in the U87-MG cancer cell line differently modified TG2 localization and expression levels. In the cells treated with free FA, TG2 appeared expressed both in the cytosol and in the nucleus, while the treatment with FA-NLCs showed that the protein is exclusively localized in the cytosol, exerting its pro-apoptotic effect. Therefore, our data suggest that FA loaded in NLCs could represent a promising natural agent for supplementing the current anti-cancer drugs used for the treatment of GBM. Full article

A new chemiluminescence immunoassay method (CLIA) for detecting IgA anti-transglutaminase (atTG IgA) in celiac disease (CD) has prompted inquiries into its diagnostic performance. We conducted a systematic review and meta-analysis comparing CLIA with traditional enzyme-linked immunosorbent assay (ELISA) and fluorescence enzyme immunoassay (FEIA). We searched PubMed, Medline, and Embase databases up to March 2024. The diagnostic references were intestinal biopsy and ESPGHAN guidelines. We calculated the sensitivity and specificity of atTG IgA assessed by CLIA and the odds ratio (OR) between the assays. Eleven articles were eligible for the systematic review and seven for the meta-analysis. Sensitivity and specificity of atTG IgA CLIA-assay were 0.98 (95% CI, 0.95–0.99) and 0.97 (95% CI, 0.94–0.99), respectively. The sensitivity of atTG IgA antibody detection did not significantly vary across the three assay modalities examined (CLIA vs. ELISA OR: 1.08 (95% CI, 0.56–2.11; p = 0.8); CLIA vs. FEIA OR: 6.97 (95% CI, 0.60–81.03; p = 0.1). The specificity of atTG IgA assessed by FEIA was higher than for CLIA (OR 0.17 (95% CI, 0.05–0.62); p < 0.007). According to the systematic review, normalization of atTG IgA levels in CD patients following a gluten-free diet was delayed when using CLIA compared to ELISA and FEIA methods. Conflicting findings were reported on the antibody threshold to use in order to avoid biopsy confirmation. Full article