Enzyme-linked Immunosorbent Assay Research Articles

Correlation between PDGF-BB and M1-type macrophage in inflammatory bowel disease: a case-control study

BackgroundInflammatory bowel disease (IBD) is a chronic disease in which macrophages play an important role in its pathogenesis. Platelet-derived growth factor-BB (PDGF-BB) secreted by macrophages is involved in the repair of vascular endothelial injury during inflammatory reactions.MethodsThe expression levels of M1 macrophages and PDGF-BB in serum and colonic mucosa of 30 patients with Crohn’s disease (CD) and 30 patients with ulcerative colitis (UC) were measured using enzyme-linked immunosorbent assays and immunohistochemistry. Logistic regression was used for univariate and multivariate analyses, and receiver operating characteristic curves were used to evaluate diagnostic value. Associations were evaluated using Spearman correlation analysis.ResultsThe expression of serum PDGF-BB and M1 macrophages with positive CXCL9 expression in patients with active-stage IBD [206.55(160.41,262.90)and 337.30(217.73,472.28) pg/ml] was higher than that in patients with remission stage [153.42(107.02,219.68)and 218.37(144.49,347.33)pg/ml] and controls [156.19(91.16,216.08)and 191.20(121.42,311.76)pg/ml](P < 0.05). The expression of PDGF-BB, CD86, and CXCL9 in the colon of patients with active-stage IBD [0.380(0.266,0.542) 0.663(0.480,0.591) and 0.564(0.378,0.765) /µm2] was higher than that in the remission stage [0.308(0.214,0.420), 0.376(0.206,0.591) and 0.413(0.275,0.570) /µm2] and controls [0.265(0.185,0.384), 0.416(0.269,0.534) and 0.497(0.415,0.642) /µm2] (P < 0.05). A positive correlation was observed between CD86 and PDGF-BB, and CXCL9 and PDGF-BB levels in patients with IBD (P < 0.05). CD86 and PDGF-BB in the colonic mucosa were independent risk factors for active IBD, and the area under the curve for their combined diagnosis was 0.754 (95%CI: 0.654–0.852, P < 0.05).ConclusionsPDGF-BB was associated with M1 macrophages and has a potential diagnostic value for active IBD.Trial registrationNot applicable.

Comparative analysis of bone health and glycemic control in postmenopausal women: insights from clinical data

Abstract Background Menopause leads to significant physiological and biochemical alterations that impact different aspects of health, such as bone mineral metabolism and glycemic control. It is imperative to comprehend these alterations in order to identify potential health hazards and develop preventative measures. This comparative cross-sectional study aimed to evaluate bone-associated markers and glycemic control indices in postmenopausal women (PMP). Methods A sample of 100 women aged 30 years or older was randomly selected and categorized into three groups: PMP (n = 60), premenopausal (PRM) (n = 20), and women in the reproductive age group (RWA) (n = 20). Venous blood samples were collected, and fasting blood glucose (FBG), calcium, phosphorus, uric acid, and alkaline phosphatase (ALP) levels were analyzed using standard laboratory techniques. The insulin and estrogen levels were evaluated using enzyme-linked immunosorbent assay (ELISA), and Homeostasis model assessment of insulin resistance (HOMA-IR), Quantitative insulin sensitivity check index (QUICKI), and Fasting insulin resistance index (FIRI) were calculated. A significance level of p &lt; 0.05 was used in the statistical analysis conducted using SPSS version 25.0. Results The PMP and PRM groups showed significantly greater mean values of FBG, insulin, and HOMA-IR when compared to RWA (p &lt; 0.05), while these groups had lower QUICKI levels. Similar trends were noted for bone-associated markers, with significant differences among the groups (p &lt; 0.05). The correlation results showed that QUICKI had positive correlations with both ALP and phosphorus, while FBG and FIRI had negative connections with both. Conclusion Postmenopausal women demonstrated higher levels of glycemic control indices and bone-related markers compared to premenopausal and reproductive-age women, indicating potential risks for osteoporosis and obesity.

TNF-α Regulated Bidirectional Interaction Between Bone Marrow Mesenchymal Stem Cells and Articular Chondrocytes.

Articular chondrocytes (ACs) secrete a variety of extracellular matrix components to maintain the functions of articular cartilage. Degeneration of ACs leads to the degeneration of articular cartilage and consequently to osteoarthritis. The secretion of bone marrow mesenchymal stem cells (BMSCs) is capable of protecting ACs from degeneration, and thus BMSCs are widely applied to treat osteoarthritis. This study aims to explore whether BMSCs and ACs will affect the functions of each other through their secretions in the context of osteoarthritis. BMSCs and ACs isolated from rabbits were identified using flow cytometry and immunocytochemistry. Conditioned medium of BMSCs and ACs treated with 0, 5, 10, 20, and 40 ng/ml of tumor necrosis factor-alpha (TNF-α) were collected and used to treat ACs and BMSCs, respectively. The viabilities of ACs and BMSCs treated with condition medium were assessed using a Cell Count Kit-8 (CCK-8) kit. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunoblotting, and enzyme-linked immunosorbent assay (ELISA) methods were employed to evaluate the relative expression levels of genes and proteins, as well as the cytokine concentrations in the supernatant. Immunofluorescence and flow cytometry results indicated that the purity of isolated cells exceeded 95%. CCK-8 analysis showed that 6 hours of treatment with a conditioned medium did not affect the viability of BMSCs and ACs. However, treatment for 12 hours or longer significantly increased the viability of BMSCs (p < 0.05) and significantly decreased the viability of ACs (p < 0.01). RT-qPCR results demonstrated that the relative expression levels of Runx2 (1.15-3.91), Alp (1.06-2.84), TNF (BMSCs: 0.94-2.54; ACs: 1.03-2.64), IL6 (BMSCs: 0.98-2.78; ACs: 0.96-3.71), IL17A (BMSCs: 1.08-5.91; ACs: 0.90-4.20), and IL10 (BMSCs: 0.93-2.82; ACs: 0.89-2.25) genes in conditioned medium-treated BMSCs and ACs were dose-dependently elevated (p < 0.001) by TNF-α treatment. Immunoblotting analysis revealed that the expression levels of RUNX2 (0.53-0.86) and ALP (0.49-0.85) proteins were also dose-dependently elevated (p < 0.001) by TNF-α treatment. ELISA results showed similar TNF-α dose-dependent increases (p < 0.001) in the supernatant concentrations of pro-inflammatory cytokines TNF-α (BMSCs: 36.90 ± 0.75 to 199.38 pg/ml; ACs: 29.76 to 293.99 pg/ml), interleukin (IL)-6 (BMSCs: 4.96-48.24 pg/ml; ACs: 6.12-38.15 pg/ml), IL-17 (BMSCs: 3.06-28.99 pg/ml; ACs: 3.08-28.51 pg/ml), as well as the anti-inflammatory cytokine IL-10 (BMSCs: 6.34-65.02 pg/ml; ACs: 5.30-34.85 pg/ml). Together, these results indicate a TNF-α-regulated bidirectional interaction between BMSCs and ACs, deepening our understanding of the pathogenesis of osteoarthritis and aiding in its prevention and treatment.

Development of validated ultrasensitive enzyme-linked immunosorbent assay method of human epidermal growth factor and in vivo pharmacokinetic study

Development of validated ultrasensitive enzyme-linked immunosorbent assay method of human epidermal growth factor and in vivo pharmacokinetic study

Invastigate The Serum Level of Interleukin-4 In Patients with Chronic Rhinosinositis with Nasal Polyps and Healthy Individuals

Objective: Interleukin-4 plays several biological roles, including stimulating the proliferation of activated B and T cells and differentiating B cells into plasma cells. This study aimed to determine whether the etiology of chronic rhinosinusitis with nasal polyps (CRSwNPs) is associated with serum levels of interleukin-4. Patients and Methods: a blood samples from 35 Iraqi patients—21 men and 14 women—who had nasal polyps as well as a control group of 35 healthy volunteers were collected. The serum was then separated in a plain tube, and an enzyme-linked immunosorbent assay method was used to determine the optical density of Interleukin-4 in the serum (ELISA) Results: Serum levels of interleukin-4 were significantly higher in the patient group compared to the control group (P &lt; 0.001). There was no significant correlation between serum interleukin-4 levels and age or gender in either group. Conclusions: Individuals who have NPs had elevated degrees of IL-4

Assessment of urine calprotectin and YKL-40 levels in urinary tract infection diagnosis in children under 2 years of age

Urinary tract infections (UTI) are common cause of fever in children. They often manifest with non-specific symptoms. New markers assessed in non-invasively collected urine are needed for quick and reliable diagnosis. This study evaluates the usefulness of calprotectin and YKL-40 in the diagnosis of febrile UTIs among children. 67 children aged between 1 month and 2 years took part in the study. The study group (group UTI) consisted of children with the first episode of febrile UTI (n = 20). Two control groups were created: group F - febrile children with other than UTI cause of the fever (n = 36), group H - healthy children (n = 11). Concentrations of calprotectin and YKL-40 in urine samples were determined with the use of the enzyme-linked immunosorbent assay method (ELISA). In the UTI group, the average calprotectin concentration was 1273.3 ng/ml, which was significantly higher than in group F (35.4 ng/ml) and in group H (8.0 ng/ml) (p < 0.001). The YKL-40 concentration in the UTI group was 15,542.1 pg/ml, which was also significantly higher than in group F (138.1 pg/ml) and in group H (162 pg/ml) (p < 0.01). These findings suggest that urinary YKL-40 and calprotectin could be useful markers in the differential diagnosis of febrile UTI among children.

Semaphorin 5A promotes Th17 differentiation via PI3K-Akt-mTOR in systemic lupus erythematosus

BackgroundPreviously, we reported that serum Semaphorin 5 A (Sema5A) levels were increased in systemic lupus erythematosus (SLE) patients compared with healthy controls (HC), and elevated Sema5A correlated with disease activity and lupus nephritis in SLE patients. In this study, we aimed to further understand the role of Sema5A in promoting Th17 cells differentiation in SLE.MethodsSema5A, interferon gamma (IFN-γ), interleukin 4 (IL-4), interleukin 17 A (IL-17 A) and interleukin 10 (IL-10) were measured by Enzyme Linked Immunosorbent Assay (ELISA). RNA and protein were isolated from peripheral blood mononuclear cells (PBMCs) in SLE patients and HC. Expression of PlexinA1 and PlexinB3 were measured by quantitative RT-PCR (qRT-PCR) and Western Blot. Th cell subsets were detected by flow cytometry. Treatment with recombinant human Sema5A (rhSema5A) and small interfering RNA (siRNA) were employed to examine the in vitro effect of Sema5A in CD4+T cell differentiation in SLE patients.ResultsIL-17 A elevated in SLE patients and positively correlated with Sema5A. PlexinA1 was upregulated and mainly expressed in CD4+ T cells of SLE; Sema5A treatment induced the differentiation of Th17 cells, while did not affect the Th1 and Th2 skewing. These effects were associated with an upregulation of the transcription factor RORγt by Th17 cells, but not T-bet or GATA3 in Th1 and Th2 cells, respectively. Knock down PlexinA1 regulates IL-17 A production by CD4+T cells. Functional assays showed that Sema5A-PlexinA1 axis promoted Th17 cells differentiation via PI3K/Akt/mTOR signaling.ConclusionsThese findings demonstrated that Sema5A-PlexinA1 axis acts as a key mediator on Th17 differentiation, suggesting that Sema5A might be a novel therapeutic target in SLE.

Hypoxia-treated adipose mesenchymal stem cells derived exosomes enhance the therapeutic effects on unilateral ureteral obstruction mice.

The exosomes from adipose-derived mesenchymal stem cells (AMSCs) had therapeutic effects. However, whether the exosomes derived from hypoxia-treated AMSCs could improve renal functions in unilateral ureteral obstruction (UUO) mice remains unclear. The exosomes were characterized using a transmission electron microscope and Western blot. Its size distribution was determined using the Zetasizer Nano ZS analysis system. The differentiation ability was assessed by alkaline phosphatase and oil red staining. Consequently, the function of exosomes in inhibiting inflammatory factors was evaluated using an enzyme-linked immunosorbent assay, and apoptosis inhibition was evaluated by Western blot. Finally, the function of exosomes to ameliorate kidney fibrosis using qPCR, Western blot, hematoxylin-eosin staining and Masson staining. The cultured AMSCs could differentiate into osteoblast and adipocyte. Meanwhile, the cultured AMSCs could effectively secrete the exosomes, which were characterized with around 110 nm diameter and surface marker expression. Exosomes derived from hypoxia-treated AMSCs improved renal functions in UUO mice. The mechanism exploration revealed that exosomes could decrease the TNF-α and IL-6 and inhibit cell apoptosis. Finally, the fibrosis-associated protein was reversed, and the renal dysfunctions were ameliorated in UUO mice. The exosomes derived from the hypoxia-treated AMSCs have a better effect than those from normal AMSCs in ameliorating renal dysfunctions in UUO mice.

Intestinal fatty acid binding protein is associated with coronary artery disease in long-term type 1 diabetes—the Dialong study

BackgroundIndividuals with type 1 diabetes are at increased risk of accelerated atherosclerosis, causing coronary artery disease (CAD). The underlying mechanisms remain unclear, but new theories proposed are damage of gut mucosa causing leakage and translocation of gut microbiota products into the circulation, leading to inflammatory responses and atherosclerosis. We therefore aimed to study the associations between gut related inflammatory biomarkers and coronary atherosclerosis in individuals with long-term type 1 diabetes.MethodsIn this cross-sectional, controlled study of 102 participants with type 1 diabetes and 63 control subjects, we measured circulating levels of intestinal fatty acid binding protein (I-FABP), soluble cluster of differentiation 14 (sCD14), lipopolysaccharide binding protein (LBP) and interleukin 18 (IL-18) by enzyme-linked immunosorbent assay (ELISA), and further gene expression of CD14 and toll-like receptor 4 (TLR4) by real time PCR in circulating leukocytes and peripheral blood mononuclear cells (PBMCs). The participants had either established coronary heart disease (CHD) or underwent computed tomography coronary angiography (CTCA) to assess for coronary atherosclerosis, including total, calcified and soft/mixed plaque volumes.ResultsIn the diabetes group, the levels of I-FABP were significantly higher in participants with established CHD or significant stenosis on CTCA compared to the participants with normal arteries or non-significant stenosis, with median 1.67 ng/ml (interquartile range [IQR] 1.02–2.32) vs. median 1.09 ng/ml (IQR 0.82–1.58), p = 0.003. I-FABP was associated with significant coronary artery stenosis by CTCA (> 50%) or previously established CHD in the adjusted analysis (odds ratio [OR] = 2.32, 95% confidence interval [CI]: 1.09–4.95; p = 0.029). The levels of I-FABP correlated also to total coronary plaque volume (r = 0.22, p < 0.05). This association remained significant after adjusting for age, sex, persistent albuminuria, eGFR, statin treatment, diabetes duration and mean time-weighted variables; HbA1c, LDL-cholesterol and systolic blood pressure (OR = 1.97, 95% CI: 1.28–3.01; p = 0.002).ConclusionsIn this cohort of individuals with long-term type 1 diabetes I-FABP associated significantly with coronary artery stenosis, suggesting a potential role of gut mucosa damage in the process of atherosclerosis in type 1 diabetes.

Recurrent BMP4 variants in exon 4 cause non-HFE-associated hemochromatosis via the BMP/SMAD signaling pathway

BackgroundHereditary hemochromatosis (HH) is an iron overload disorder and can be caused by variants in non-HFE genes in Chinese patients. However, there is still a considerable proportion of patients suffering from unexplained iron overload. In our previous study, we had identified the p.R269Q variant in exon 4 of the Bone morphogenetic protein 4 (BMP4) gene in Chinese patients with unexplained primary iron overload by Whole Exome sequencing, and then the BMP4 p.H251Y variant was identified by Sanger sequencing in a Chinese patient with secondary iron overload. Our study aimed to explore the pathogenicity and underlying mechanism of BMP4 p.H251Y and BMP4 p.R269Q variants in patients with iron overload.MethodsSanger sequencing was conducted to identify the novel variants in the BMP4 gene of patients with unexplained iron overload. MRI and liver biopsy were used to display iron overload in the liver of the patient harboring the BMP4 p.H251Y variant. The BMP4 and hepcidin levels in BMP4 knockdown and BMP4 variant cells were examined by enzyme-linked immunosorbent assay. The effects of BMP4 p.H251Y and BMP4 p.R269Q variants on the hepcidin-regulation pathway were studied.ResultsOne of 54 HH patients (1.85%) harbored the BMP4 p.R269Q variant. One of 148 patients (0.68%) with secondary hemochromatosis harbored the BMP4 p.H251Y variant, and these two variants were not found in 100 Chinese general population. For the patient harboring the BMP4 p.H251Y variant, abdominal MRI and Perl's staining of liver tissue displayed iron overload in the liver. Cells transfected with the BMP4 p.H251Y and p.R269Q variants showed down-regulation of hepcidin level and BMP/SMAD pathway compared with cells transfected with the wild-type BMP4 vector.ConclusionThe BMP4 p.H251Y and p.R269Q variants can downregulate hepcidin levels by inhibiting the BMP/SMAD axis, suggesting they may play pathogenic roles in iron overload.

Transdermal Delivery of Tofacitinib Citraste via Mannose-Decorated Transferosomes Loaded with Tofacitinib Citrate in Arthritic Joints.

Transdermal drug delivery systems are a promising option for the treatment of rheumatoid arthritis (RA) because they can lower systemic adverse effects of immunosuppressants. Janus kinase (JAK) inhibitors were found to be effective for the treatment of RA by inhibiting the JAK-STAT pathway and preventing autoimmune joint destruction. The aim of this study is to deliver tofacitinib (a JAK 1 and 3 inhibitor) through mannose-decorated transferosomes (MDTs) directly to inflamed joints. Transferosomes are composed of phospholipids, Cremophor A25, PEG400, Labrafac lipophile, and oleic acid to enhance the permeation of tofacitinib and control nanovesicle size (∼70-200 nm). Permeation through rat skin was evaluated, where the skin permeation of MDTs (Q24: 38.8 ± 9.82 μg/cm2) and flux (0.5311 ± 0.072 μg/cm2/h) were significantly greater than those of the uncoated transferosomes (Q24 of T1: 1.522 ± 0.329 μg/cm2, Q24 of T2: 3.5002 ± 0.998 μg/cm2, and Q24 of T3: 18.226 ± 5.25 μg/cm2). In addition, MDTs seem to permeate the skin intact, as shown by the transmission electron microscopy (TEM) images of the recipient buffer removed from the Franz diffusion cell. A histopathology assay was performed during the in vivo evaluation of MDTs in an arthritic rat model, in which, significantly less inflammation was observed when MDTs were applied directly to the joint compared to when applied to the dorsal skin and untreated arthritic joints. Furthermore, significantly lower tumor necrosis factor-α (TNFα), IL-6, and IL-1β levels (P < 0.05) were detected by enzyme-linked immunosorbent assay (ELISA) in homogenates of the joints treated with MDTs than in untreated arthritic joints. In conclusion, this study proposed effective MDTs that could deliver tofacitinib directly to inflamed joints possibly by targeting the macrophages circulating in the proximity of the site of inflammation.

Exploring the Role of Adipose Tissue Dysregulation in Vitiligo Pathogenesis: A Body Composition Analysis.

Vitiligo is a commonly acquired pigmentary disease characterized by the progressive destruction of melanocytes. The role of adipose tissue in vitiligo remains unclear. To investigate the role of adipose tissue in the pathogenesis of vitiligo, 50 patients newly diagnosed with vitiligo and 40 age- and sex-matched healthy controls were enrolled, and fat mass and distribution using dual-energy X-ray absorptiometry (DXA) and serum adipokine levels using enzyme-linked immunosorbent assay were assessed. The trunk and whole-body fat mass and the trunk/limb fat mass ratio were significantly higher in patients with vitiligo. Higher leptin and resistin levels and lower adiponectin levels were observed. Positive correlation of the trunk fat mass index with vitiligo area scoring index scores (r = 0.38, p = 0.0071) was found. Additionally, plasma adiponectin levels were negatively correlated with vitiligo disease activity scores (r = -0.307, p < 0.05). Receiver operating characteristic curve analysis showed that the percentage of limb lean tissue mass, trunk/limb fat mass ratio, and serum adipokine levels achieved a high area under the curve score for distinguishing patients with vitiligo from healthy controls. In conclusion, the incidence of central obesity and adipokine dysregulation was higher in patients with vitiligo. The potential role of adipose tissue in the pathogenesis of vitiligo should be emphasized.

Mechanism of vitamin D deficiency involvement in the development of pulmonary arterial hypertension related to monocrotaline-induced connective tissue disease in rats

Objective: To investigating the impact of vitamin D (VitD) deficiency on the jagged 1 protein (Jagged1)/Notch3 signaling pathway in the pulmonary arteries of rats with monocrotaline (MCT)-induced connective tissue disease (CTD)-related pulmonary arterial hypertension (PAH) and to explore the pathological and molecular mechanisms of VitD involvement in the development of CTD-PAH. Methods: Twenty-four 7-week-old male Wistar rats were divided into a normal diet group and a VitD-free diet group using random number table, with 12 rats in each group. After 5 weeks of feeding, the rats were further randomly divided into saline and MCT groups, forming group A (normal diet+saline), group B (normal diet+MCT), group C (VitD-free+saline), and group D (VitD-free+MCT), with 6 rats in each group, and the rats were continued to be fed for another 4 weeks. The MCT group was injected with MCT solution subcutaneously on the back of the neck to construct the CTD-PAH model, and the saline group was injected with an equal amount of saline as a control. At the end of the experiment, blood samples were collected from all rats, and the serum 1, 25-dihydroxy vitamin D levels were determined by enzyme-linked immunosorbent assay. Hemodynamic measurements were performed on all rats, recording and calculating the mean values of right ventricular pressure and pulmonary artery pressure. Lung tissues and pulmonary arteries of all rats were stained with hematoxylin-eosin staining, the inner and outer diameters of pulmonary vessels were measured, and the percentage of pulmonary artery medial layer thickness was calculated. The right ventricular hypertrophy index was determined by weighing the parts of the heart and calculating the ratio. The apoptosis of pulmonary artery smooth muscle cells was assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. The expression levels of Jagged1, Notch3, and hairy and enhancer of split-1 (Hes1) mRNA and protein in the pulmonary arteries were analyzed by reverse transcription quantitative polymerase chain reaction and Western blotting. Results: The serum 1, 25-dihydroxy vitamin D levels in group D were significantly lower than those in groups A, B, and C [(51.01±0.96) μg/L, (65.15±1.83), (57.49±0.28), (54.52±2.87) μg/L, respectively, all P<0.05]. The right ventricular pressure, pulmonary artery systolic pressure, pulmonary artery diastolic pressure, and mean pulmonary artery pressure in Group D rats were significantly higher than those in groups A, B and C (all P<0.05). The percentage of pulmonary artery medial layer thickness in group D (56.28%±9.51%) was significantly higher than that in Groups A (21.28%±1.89%), B [22.72% (22.16%, 30.10%)], and C (38.73%±7.34%) (all P<0.05); the right ventricular hypertrophy index in group D (45.74%±12.68%) was significantly higher than that in groups A (21.78%±2.80%), B (40.93%±9.10%), and C (23.71%±1.22%) (all P<0.05). The apoptosis of pulmonary artery smooth muscle cells in group D (0.07±0.01) was significantly lower than that in groups A (1.00±0.08), B (0.17±0.02), and C (0.49±0.07) (all P<0.05). The levels of Jagged1, Notch3, and Hes1 mRNA and protein in Group D were all significantly higher than those in Groups A, B, and C (all P<0.05). Conclusions: VitD deficiency is involved in the pathophysiological mechanisms of CTD-PAH development. This occurs by upregulating the expression of the Jagged1/Notch3/Hes1 pathway in the pulmonary arteries of rats with MCT-induced PAH. The consequences include pulmonary artery thickening, increased pulmonary artery pressure, increased right ventricular pressure, right ventricular hypertrophy, and decreased apoptosis of pulmonary artery cells.

Differential murine responses to Schistosoma mansoni eggs in the liver and small intestine lead to downmodulation of hepatic but not intestinal periovular granulomas.

To control schistosomiasis mansoni, it is important to attempt preventing the worms' egg-induced pathology in the liver and limiting pathogen transmission following egg exit from the intestines to the exterior. Therefore, the present study aimed to clarify the reasons behind the decades-long riddle of periovular granulomas downmodulation in the liver, but not the small intestine, with the progression of murine schistosomiasis mansoni. Outbred female CD-1 mice were percutaneously exposed to 15 Schistosoma mansoni cercariae. The liver and small intestine were collected from mice harboring a minimum of a worm couple at 8, 12, 16, and 20 weeks post-infection, assessed for egg counts/g and histopathological changes, and used to prepare Triton X-100 extracts. Content of cytokines, saturated and unsaturated fatty acids, triglycerides, cholesterol, reactive oxygen species, and uric acid per mg tissue extract proteins were evaluated using capture enzyme-linked immunosorbent assays, gas chromatography-flame ionization detector, and standard commercially available reagents, respectively. Examination of hematoxylin-eosin-stained tissue sections confirmed the decrease in size and changes in cellular composition of periovular granulomas in the liver but not the small intestine, associated with wide differences in released cytokines types and amounts, and content of the bioactive lipids, arachidonic and docosahexaenoic acids, reactive oxygen species, and uric acid. The results together disclosed that the downmodulation of hepatic, but not the small intestine, circumoval granulomas with the progression of murine S. mansoni naturally results from site- and tissue- specific immunological and biochemical responses to the egg-derived antigens and molecules and suggested that the intestines appear to harbor immune-privileged sites.

Association of Chronic Urticaria With Hepatitis C Infection In Female Patients Visiting Tertiary Care Hospital In Pakistan

Objective: To evaluate association of chronic urticaria with hepatitis C infection in female patients. Study Design: comparative cross-sectional study. Place and Duration of Study: Fauji Foundation Hospital Rawalpindi. One-year duration (Jan 2020-Jan 2021). Methodology: One Hundred female patients (group 1) of age 60 years or less with chronic urticaria were enrolled in this study. Patients with chronic liver disease or signs of hepatic decompensation were excluded. Equal number of age and gender matched patients not having urticarial symptoms were selected as group 2. Anti-Hepatitis C Virus Antibody (HCV ab) was evaluated by Enzyme linked immunosorbent assay (ELISA). Disease severity was calculated by Urticaria activity score 7(UAS 7). Association of HCV infection with chronic urticaria and its severity was analyzed by applying appropriate statistical tests. Results: Anti HCV antibody positivity frequency was 14/100(14%) in group 1 and 4/100(4%) in group 2 (p value: 0.013; significant). Mean ages in group 1 and 2 were 39.88+1.38 and 38.79+1.35 years respectively (p value: 0.57; insignificant). Mean UAS 7 calculated in the HCV negative Urticaria cases was 6.73+2.94 while it was significantly higher in HCV positive group i.e. 8.78+6.459 (p value 0.01). Frequency of HCV positivity in patients with well controlled urticaria and mild urticaria was 6/59(10.0%) and 5/36(13.8%) respectively while in moderate urticaria was 3/5(60%) which was significantly higher (p value 0.04). Conclusion: There is a significant association of chronic urticaria with anti HCV antibody positivity. HCV positive patients have more severe disease as compared to HCV negative patients.

Inhibition of iron-induced cofilin activation and inflammation in microglia by a novel cofilin inhibitor.

Neuroinflammatory conditions linked to iron dysregulation pose significant challenges in neurodegenerative diseases. Iron-loaded microglia are observed in the brains of patients with various neuroinflammatory conditions, yet how iron overload affects microglial function and contributes to various neuroinflammatory processes is poorly understood. This invitro study elucidates the relationship between excess iron, cofilin activation, and microglial function, shedding light on potential therapeutic avenues. Iron overload was induced in Human Microglial Clone 3 cells using ferrous sulfate, and the expressions of ferritin heavy chain, ferritin light chain, divalent metal transporter 1, cofilin, p-cofilin, nuclear factor-κB (NF-κB), and various inflammatory cytokines were analyzed using real-time quantitative polymerase chain reaction, immunocytochemistry, Western blotting, and enzyme-linked immunosorbent assay. Results revealed a notable increase in cofilin, NF-κB, and inflammatory cytokine expression levels following excess iron exposure. Moreover, treatment with deferoxamine (DFX), a known iron chelator, and a novel cofilin inhibitor (CI) synthesized in our laboratory demonstrate a mitigating effect on iron-induced cofilin expression. Furthermore, both DFX and CI exhibit promising outcomes in mitigating the inflammatory consequences of excess iron, including the expression of pro-inflammatory cytokines and NF-κB activation. These findings suggest that both DFX and CI can potentially alleviate microglia-induced neuroinflammation by targeting both iron dysregulation and cofilin-mediated pathways. Overall, this study provides valuable insights into iron-induced cofilin activation and microglial activation, offering avenues for potential targeted therapies for neuroinflammatory conditions associated with iron and cofilin dysregulation in neurodegenerative diseases.

The overexpression of eIF4E decreases oxytocin levels and induces social-cognitive behavioral disorders in mice.

Overexpression of the eukaryotic initiation factor 4E (eIF4E) gene has been associated with excessive stereotypic behaviors and reduced sociability, which manifest as autism-like social cognitive deficits. However, the precise mechanisms by which eIF4E overexpression induces insufficiently these autism-like behaviors and the specific brain regions implicated remain insufficiently understood . Oxytocin, a neurotransmitter known for its role in social behavior, has been proposed to modulate certain autism-related symptoms by influencing microglial function and attenuating neuroinflammation. Nonetheless , the contributions of the hippocampus and oxytocin in the content of eIF4E overexpression-induced autistic behaviors remain elucidated . To investigate this issue,esearchers utilized the three-chamber social interaction test, the open field test, and the Morris water maze to evaluate the social cognitive behaviors of the two groups of mice. Additionally, enzyme-linked immunosorbent assay (ELISA), immunofluorescence, Western blotting, and RT-qPCR were employed to quantify oxytocin levels and assess hippocampal microglial activation.The results indicate that overexpression of eIF4E in mice is associated with significant impairments in social cognition, alongside pronounced marked hyperactivation of hippocampal microglia.Significance statement Autism spectrum disorder (ASD) encompasses a range of neurodevelopmental disorders characterized by social cognitive impairment.Research has indicated a correlation between the overexpression of the eukaryotic initiation factor 4E (eIF4E) gene and autism-like social cognitive impairment.Oxytocin (OXT), a neurotransmitter, plays a role in regulating hippocampal microglial activity and attenuating neuroinflammation. This modulation may impact social cognition in individuals with autism.Nevertheless,it remains unclear whether there is an involvement of the hippocampus and oxytocin in autism-like social cognitive impairments due to eIF4E overexpression. The present study suggests that overexpression of eIF4E may induce hyperactivation of microglia and contribute to social cognitive impairment by decreasing oxytocin levels in the hippocampus.These findings offer molecular insights into the manifestation of autism-like behavior resulting from eIF4E overexpression and may guide future clinical interventions.

Relationship Between Serum Levels of Oxidized Lipoproteins, Circulating Levels of Myeloperoxidase and Paraoxonase 1, and Diet in Young Subjects with Insulin Resistance.

Oxidized low-density lipoproteins (ox-LDLs) are involved in atherosclerotic plaque formation and progression and have been linked to insulin resistance (IR). Myeloperoxidase is a potent oxidant of lipoproteins related to atherogenic risk. High-density lipoproteins (HDLs) are considered antioxidants due to their association with paraoxonase 1 (PON1). However, HDL can also be oxidized (ox-HDL), and its relationship with IR has not been described. This study evaluated the relationship between circulating levels of myeloperoxidase and paraoxonase 1, diet, and serum levels of ox-LDL and ox-HDL in young people with IR. This cross-sectional study examined 136 young subjects (67 and 69 with and without insulin resistance, respectively). Serum levels of ox-LDL, ox-HDL, myeloperoxidase, and PON1 were quantified using an enzyme-linked immunosorbent assay. The nutritional dietary content of the foods was determined with a food frequency questionnaire, which was analyzed with Nutrimind 2013 software. Serum ox-HDL levels were higher in young subjects without IR than those with IR (p = 0.031). Women with IR presented increased ox-LDL levels compared with women without IR (p = 0.012) and men with IR (p < 0.001). In the IR group, serum ox-LDL levels were negatively correlated with total cholesterol, triglycerides, and LDL-C, whereas the correlation was positive in the insulin-sensitive group. Consumption of vitamins B1 and B2 was related to increased HDL-C levels, while higher ox-LDL levels were related to vitamin K intake. In addition, low energy consumption and phosphorus increased PON1 levels. The results suggest that insulin resistance in young women may promote lipoprotein oxidation, and the intake of B complex vitamins may have an antiatherogenic effect.

Neuroprotective and inflammatory biomarkers in pediatric drug-resistant epilepsy: Interplay between GDNF, IL-1β and vitamin D 25-OH

Drug-resistant epilepsy in pediatric patients is associated with neuroinflammation and neurodegeneration. Vitamin D 25-OH exerts neuroprotective effects, while glial cell line-derived neurotrophic factor (GDNF) and the proinflammatory cytokine interleukin-1β (IL-1β) are implicated in the mechanisms of neuroinflammation and epileptogenesis. The aim of this study was to investigate the relationship between vitamin D 25-OH, IL-1β, and GDNF levels with seizure severity and frequency in children with drug-resistant epilepsy. A cross-sectional study was conducted at Adam Malik Hospital, Medan, Indonesia, among children with drug-resistant epilepsy. Vitamin D 25-OH, IL-1β and GDNF levels were measured using enzyme-linked immunosorbent assay (ELISA). Epilepsy severity was assessed using the Hague Seizure Severity Scale (HASS), while seizure frequency was assessed using the Global Assessment of Severity of Epilepsy (GASE). The present study identified a significant correlation between GDNF levels and epilepsy severity, as measured by the HASS score (r=0.318; p=0.006). However, no significant correlation was observed between vitamin D 25-OH or IL-1β levels and epilepsy severity or seizure frequency (p&gt;0.05). IL-1β levels correlated significantly with GDNF levels (r=0.525; p=0.001), but IL-1β did not directly correlate with seizure frequency or epilepsy severity. In conclusion, GDNF levels significantly correlated with epilepsy severity, suggesting that GDNF may serve as a potential biomarker for assessing epilepsy severity. However, further studies investigating the role of GDNF as a potential neurotrophic factor in the pathophysiology of epilepsy and its possible application as a therapeutic target are important.

Evidence of Site-Specific Mucosal Autoantibody Secretion in Rheumatoid Arthritis.

Anti-citrullinated protein antibodies (ACPA) have been detected in sputum and saliva, indicating that anti-modified protein antibodies (AMPA) can be produced at mucosal sites in patients with rheumatoid arthritis (RA). However, the body's largest mucosal compartment, the gut, has not yet been examined. We therefore investigated the presence of several AMPA(ACPA, anti-carbamylated protein antibodies [anti-CarP], and anti-acetylated protein antibodies [AAPA]) at different mucosal sites, including the intestinal tract. Paired fecal/ileal wash, saliva, and serum samples of patients with RA and healthy volunteers were collected in two independent cohorts. Data involving feces were replicated in a third cohort. In these secretions, AMPA were analyzed using in-house enzyme-linked immunosorbent assay with unmodified peptides as control. In fecal samples, total IgA and anti-Escherichia coli IgA were measured. ACPA, anti-CarP, and AAPA IgA were measurable in saliva of seropositive patients with RA (prevalence 9%-40%). No AMPA could be detected in feces. IgA was present because total IgA and anti-E. coli IgA were detectable in feces of ACPA-positive patients with RA and healthy donors. Results were confirmed in another cohort using colonoscopically collected ileal wash samples. Our study shows the presence of ACPA, anti-CarP, and AAPA IgA in saliva of ACPA-seropositive patients with RA. However, no AMPA could be detected in feces/ileal wash samples of these patients, although our assays were able to measure other antigen-specific antibodies. These data suggest that mucosal autoantibody secretion may occur in the oral mucosa of patients with RA, whereas no evidence could be found for this process in the lower gastrointestinal tract.