Assessment of a combination of plasma anti-histone autoantibodies and PLA2/PE ratio as potential biomarkers to clinically predict autism spectrum disorders.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by deficiencies in social interaction and repetitive behaviors. Multiple studies have reported abnormal cell membrane composition and autoimmunity as known mechanisms associated with the etiopathogenesis of ASD. In this study, multiple regression and combined receiver operating characteristic (ROC) curve as statistic tools were done to clarify the relationship between phospholipase A2 and phosphatidylethanolamine (PE) ratio (PLA2/PE) as marker of lipid metabolism and membrane fluidity, and antihistone-autoantibodies as marker of autoimmunity in the etiopathology of ASD. Furthermore, the study intended to define the linear combination that maximizes the partial area under an ROC curve for a panel of markers. Forty five children with ASD and forty age- and sex-matched controls were enrolled in the study. Using ELISA, the levels of antihistone-autoantibodies, and PLA2 were measured in the plasma of both groups. PE was measured using HPLC. Statistical analyses using ROC curves and multiple and logistic regression models were performed. A notable rise in the area under the curve was detected using combined ROC curve models. Additionally, higher specificity and sensitivity of the combined markers were documented. The present study indicates that the measurement of the predictive value of selected biomarkers related to autoimmunity and lipid metabolism in children with ASD using a ROC curve analysis should lead to a better understanding of the pathophysiological mechanism of ASD and its link with metabolism. This information may enable the early diagnosis and intervention.

Estrogenization of insulin by catecholestrogen produced high affinity autoantibodies and altered the normal function of insulin in type 1 diabetes.

AIMS: Insulin (Ins) covalently modified by catecholestrogens (CEs) was commonly found in diabetic patients who have developed insulin resistance. Estrogenization of insulin altered its molecular function and effect carbohydrates metabolisms in these patients. Insulin resistance is a common phenomenon in diabetes but the exact mechanism remains unknown. In this study, binding specificity and affinity of autoantibodies against estrogenized insulin (4-hydroxyestradiol-insulin; 4-OHE2-Ins) were assayed in the serum of type 1 diabetes (T1D) patients in order to explain the phenomena behind insulin resistance. MATERIALS AND METHODS: Specificity and affinity of autoantibodies from the sera of 66 T1D patients and 41 controls were analyzed by direct binding, competition ELISA and quantitative precipitin titration. Insulin was also estimated in the serum of T1D patients by ELISA. KEY FINDING: Estrogenized insulin (4-OHE2-Ins) exhibited high affinity and specificity to T1D autoantibodies in comparison to Ins (p < .05) or 4-OHE2 (p < .001). Estrogenization of insulin alters its interaction with the insulin receptor (IR). The affinity constant of 4-OHE2-Ins with the T1D autoantibodies was found to be 1.41 × 10-7 M. SIGNIFICANCE: Estrogenization of insulin by catecholestrogen makes these molecules highly antigenic and produced high-affinity autoantibodies in T1D patients. As a result, patients develop insulin resistance and presented this molecule as a potential biomarker for T1D.

Systematic Assessment of Immune Marker Variation in Type 1 Diabetes: A Prospective Longitudinal Study.

Immune analytes have been widely tested in efforts to understand the heterogeneity of disease progression, risk, and therapeutic responses in type 1 diabetes (T1D). The future clinical utility of such analytes as biomarkers depends on their technical and biological variability, as well as their correlation with clinical outcomes. To assess the variability of a panel of 91 immune analytes, we conducted a prospective study of adults with T1D (<3 years from diagnosis), at 9-10 visits over 1 year. Autoantibodies and frequencies of T-cell, natural killer cell, and myeloid subsets were evaluated; autoreactive T-cell frequencies and function were also measured. We calculated an intraclass correlation coefficient (ICC) for each marker, which is a relative measure of between- and within-subject variability. Of the 91 analytes tested, we identified 35 with high between- and low within-subject variability, indicating their potential ability to be used to stratify subjects. We also provide extensive data regarding technical variability for 64 of the 91 analytes. To pilot the concept that ICC can be used to identify analytes that reflect biological outcomes, the association between each immune analyte and C-peptide was also evaluated using partial least squares modeling. CD8 effector memory T-cell (CD8 EM) frequency exhibited a high ICC and a positive correlation with C-peptide, which was also seen in an independent dataset of recent-onset T1D subjects. More work is needed to better understand the mechanisms underlying this relationship. Here we find that there are a limited number of technically reproducible immune analytes that also have a high ICC. We propose the use of ICC to define within- and between-subject variability and measurement of technical variability for future biomarker identification studies. Employing such a method is critical for selection of analytes to be tested in the context of future clinical trials aiming to understand heterogeneity in disease progression and response to therapy.

Assessment of pre and post-thymectomy myasthenia gravis.

OBJECTIVE: To evaluate transcervical and transsternal thymectomy benefits in large myasthenia gravis (MG) cohort. METHOD: We retrospectively evaluated MG patients (n = 184) who had undergone thymectomy between 2004 and 2015 at National Institute of Mental Health and Neurosciences, Bangalore (India). Myasthenia gravis foundation of America guidelines were followed to assess clinical outcome. Anti-acetylcholine receptors (AChR) antibodies, repetitive nerve stimulation (RNS) and Neostigmine tests were performed at pre and post-thymectomy stage. RESULTS: Most of the patients were fell under MG grade IIA (82 of 184, 44.56%) and grade IIB (61 of 184, 33.15%). Thymoma and thymic hyperplasia was established in 64 (34.78%) and 89 (48.37%) patients respectively. Other thymic abnormalities such thymic atrophy, cysts and lipoma were established in 31 (16.85%) patients. MG patients were treated either with transcervical (n = 79) or (n = 105) transsternal thymectomy. At the pre-thymectomy stage, the majority of the patients were positive for anti-AChR antibodies (179 of 184, 97.28%), RNS (170 of 184, 92.4%), and Neostigmine (175 of 184, 95.11%). At the post-thymectomy stage, a significant reduction observed in anti-AChR antibodies positivity (p < 0.022) and RNS positivity (p < 0.015). Overall, benefits were observed in 61.41% (113 of 184) of patients. Clinical benefits (complete stable remission, pharmacological remission, minimal manifestation, and improvement) of transcervical and transsternal thymectomy observed in 69.62% (55 of 79) and 55.24% (58 of 105) of patients respectively. MG patients with thymoma showed the least improvement compared to thymic hyperplasia. DISCUSSION: Transcervical and transsternal thymectomy showed clinical benefits, however, there was no significant difference between them.

Quantitative assessment of thyroid gland elasticity with shear-wave elastography in pediatric patients with Hashimoto's thyroiditis.

OBJECTIVE: Hashimoto's thyroiditis is the most common autoimmune thyroid disorder in the pediatric age range. Measurement of thyroid gland size is an essential component in evaluation and follow-up of thyroid pathologies. Along with size, tissue elasticity is becoming a more commonly used parameter in evaluation of parenchyma in inflammatory diseases. The aim of the current study was to assess thyroid parenchyma elasticity by shear-wave elastography in pediatric patients with Hashimoto's thyroiditis; and compare the elasticity values to a normal control group. MATERIALS AND METHODS: In this study; thyroid glands of 59 patients with a diagnosis of Hashimoto's thyroiditis based on ultrasonographic and biochemical features, and 26 healthy volunteers without autoimmune thyroid disease and thyroid function disorders, were evaluated with shear-wave elastography. Patients with Hashimoto thyroiditis were further subdivided into three categories based on gray-scale ultrasonography findings as focal thyroiditis (grade 1), diffuse thyroiditis (grade 2), and fibrotic thyroid gland (grade 3). RESULTS: Patients with Hashimoto's thyroiditis (n = 59) had significantly higher elasticity values (14. 9 kPa; IQR 12.9-17.8 kPa) than control subjects (10.6 kPa; IQR 9.0-11.3 kPa) (p < 0.001). Of the 59 patients with Hashimoto's thyroiditis, 23 patients had focal thyroiditis involving less than 50% of the gland categorized as grade 1, 24 patients had diffuse involvement of the thyroid gland categorized as grade 2, and 12 patients had marked hyperechoic septations and pseudonodular appearance categorized as grade 3 on gray-scale ultrasound. Based on elastography, grade 3 patients had significantly higher elasticity values (19.7 kPa; IQR 17.8-21.5 kPa) than patients with grade 2 (15.5 kPa; IQR 14.5-17.8 kPa) and grade 1 thyroiditis (12.8 kPa; IQR 11.9-13.1 kPa) (p < 0.05). Patients with grade 2 thyroiditis had significantly higher elasticity values than those with grade 1 thyroiditis (p < 0.05). CONCLUSION: Gray-scale ultrasound findings of heterogeneous echotexture and hypoechoic echogenicity reflect a longer duration of inflammation and may not be found in the initial stages of thyroiditis. Our results indicate that shear-wave elastography could be used to evaluate the degree of fibrosis in Hashimoto's thyroiditis.

HLA genotyping as first-line screening tool for coeliac disease in children with juvenile idiopathic arthritis.

OBJECTIVES: Coeliac disease (CD) and juvenile idiopathic arthritis (JIA) often coexist. This association warrants assessment for CD in patients with JIA. We evaluated the clinical relevance and cost-effectiveness of human leucocyte antigen (HLA) genotyping in first-line screening for development of CD in children with JIA. PATIENTS AND INTERVENTIONS: 95 patients with JIA were screened for CD using CD-specific antibodies. In case of positivity, a small intestinal biopsy was performed to confirm diagnosis. In addition, HLA genotyping was performed. 110 age-matched and sex-matched Caucasian children from the same geographical area served as controls. RESULTS: CD was diagnosed in 4 of 95 patients with JIA (4.2%), a rate significantly higher compared with controls (p<0.02) and 14 times higher than in the general population. Twenty-six patients (27.4%) had one of the variants of the risk genotypes. All four patients diagnosed with CD had a HLA-DQ2.5 genotype: one was homozygote, the remainder heterozygote. Twenty-two patients are, judging by their HLA genotypes, at risk of developing CD and require repeated serological screening. None of the 69 patients without HLA-DQ2/DQ8 genotypes had CD-specific antibodies. Screening with HLA genotyping becomes cheaper than screening without after the second determination. CONCLUSIONS: In our cohort of patients with JIA, lack of HLA-DQ2/DQ8 genotypes identified a majority not at risk of CD in whom repeated serological testing is unnecessary. Genotyping is nowadays the most efficient and cost-effective way to screen for CD risk in JIA.

Assessment of the humoral immune response to cancer.

One of the deadly hallmarks of cancer is its ability to prosper within the constraints of the host immune system. Recent advances in immunoproteomics and high-throughput technologies have lead to profiling of the antibody repertoire in cancer patients. This in turn has lead to the identification of tumour associated antigens/autoantibodies. Autoantibodies are extremely attractive and promising biomarker entities, however there has been relatively little discussion on how to interpret the humoral immune response. It may be that autoantibody profiles hold the key to ultimately uncovering neoplastic associated pathways and through the process of immunosculpting the tumour may have yielded an immune response in the early stages of malignant tumour development. The aim of this review is to discuss the utility of the autoantibody response that is elicited as a result of malignancy and discuss the advantages and limitations of autoantibody profiling. This article is part of a Special Issue entitled: Translational Proteomics.

The potential roles for novel biomarkers in rheumatoid arthritis assessment.

Comprehensive management of rheumatoid arthritis (RA) requires regular monitoring of disease activity, functional status, and structural damage to facilitate optimal patient outcomes. Tight control strategies have been successfully used in other diseases including diabetes and hypertension. Tight control requires frequent disease activity measurements in order to tailor treatment for individual patients, resulting in improved patient outcomes. Current monitoring measures used in clinical practice are largely driven by subjective evaluation of signs and symptoms, which are critical but limited by assessor variability and may not reflect true biological change in a timely manner. Research suggests that novel biomarkers may provide quantitative, objective assessments of disease activity and structural damage risk in RA, which are not captured by current measures. The simultaneous use of multiple biomarkers in a single test algorithm may provide a more comprehensive quantitative representation of the overall complex heterogeneous biology of RA. This article reviews the current management strategies for monitoring RA and the potential impact that multi-biomarker assays may have on RA assessment, which may further improve clinical outcomes.

Retroperitoneal fibroses: aetiopathogenesis and taxonomic assessment.

Retroperitoneal fibrosis (RPF) is a chronic retroperitoneal inflammatory process that can entrap the retroperitoneal structures, mainly the ureters and the great vessels. Aetiology, clinical features and diagnostic appearance in several cases are protean. A true idiopathic form is present in any cases of RPF in which no potential aetiologic condition may be identified. The pathogenesis of the idiopathic RPF appears today to be related to IgG4 autoimmune mechanisms ("hyper-IgG4 disease"). Otherwise, RPF in the presence of aortic atheromatous inflammation (atheromatous aortitis), has been included, more than twenty years ago, among the secondary forms, since this condition appears to be elicited by antigen-acting oxidized-LDL and/or ceroid, that are present within the atheromatous plaque. Aetiology of other secondary RPFs refers to medications (drug-induced), infections, traumas, malignancies. Recent advances in imaging techniques (TC, RM, 18F-FDG/PET or hybrid TC/PET), together with laboratory findings (CRP, ESR, IgG, IgG4, autoantibodies, etc), allow to identify the active phases of the inflammatory process. The review focuses on the pathogenetic features of RPFs and some issues concerning their taxonomic assessment.

Assessment of the relevance of zona pellucida antibodies in follicular fluid of in-vitro fertilization (IVF) patients.

The presence of anti-zona pellucida antibodies in the follicular fluid of 11 women who underwent in-vitro fertilization (IVF) and embryo transfer was analysed. Only infertile couples with tubal or unexplained pathologies were included in our study, which was aimed at investigating the relationship between anti-zona pellucida antibodies in follicular fluid and failed fertilization. Whether or not these antibodies were present in some or all follicles in the same patient was also investigated. Out of 55 follicular fluids analysed, 36.3% were positive to the test and no fertilization was observed in oocytes from these follicles, while 63.6% were negative, and the oocyte fertilization rate associated with these was 51.4%. The presence of anti-zona pellucida antibodies was positively correlated with the degree of fertilization failure (P < 0.001 chi 2 test).