A new ELISA for autoantibodies to steroid 21-hydroxylase.

BACKGROUND: A new ELISA for autoantibodies to steroid 21-hydroxylase (21-OH Ab) is described. METHODS: In the assay test sample autoantibodies form a bridge between 21-OH coated onto the plate well and liquid phase 21-OH-biotin. Bound 21-OH-biotin is detected by the addition of streptavidin peroxidase and colorogenic peroxidase substrate. RESULTS: Of 100 samples from patients with autoimmune Addison's disease, 86 (86%) were positive for 21-OH Ab ELISA whereas 84 (84%) were positive in an immunoprecipitation assay based on 125I-labeled 21-OH. Six (0.6%) of 928 healthy adult blood donors and 1 (2.0%) of 49 adult patients with type 1 diabetes mellitus (T1DM) were positive by ELISA. No samples from adult patients with Graves' disease (GD; n=50), celiac disease (n=29), systemic lupus erythematosis (n=9) or rheumatoid arthritis (n=20) were positive by ELISA. However, 2/51 (3.9%) children with GD, 3/69 (4.3%) children with Hashimoto's thyroiditis (HT) and 3/119 (2.5%) children with T1DM alone or associated with autoimmune thyroid disorders were ELISA positive. CONCLUSIONS: The new assay should be useful for screening patients known to be at increased risk of developing clinical autoimmune Addison's disease, in particular children with HT, GD and/or T1DM.

Assessment of autoantibodies to interferon-ω in patients with autoimmune polyendocrine syndrome type 1: using a new immunoprecipitation assay.

BACKGROUND: Measurements of autoantibodies to interferon-ω (IFN-ω) in patients with autoimmune polyglandular syndrome type 1 (APS-1) were performed using a new immunoprecipitation assay (IPA) based on 125I-labeled IFN-ω. METHODS: We have developed and validated a new IPA based on 125I-labeled IFN-ω. Sera from 78 patients (aged 3-78 years) with clinically diagnosed APS-1, 35 first degree relatives, 323 patients with other adrenal or non-adrenal autoimmune diseases and 84 healthy blood donors were used in the study. In addition, clinical features and autoimmune regulator (AIRE) genotype for the APS-1 patients were analyzed. RESULTS: Sixty-six (84.6%) of 78 APS-1 patients were positive for IFN-ω Ab using 125I-labeled IFN-ω IPA. IFN-ω Ab was the most prevalent of the six different autoantibodies tested in this group of APS-1 patients. All 66 IFN-ω Ab-positive APS-1 patients had AIRE mutations and 7 IFN-ω Ab-negative patients had no detectable AIRE mutations, whereas 3 (3.8%) patients were discrepant for IFN-ω Ab positivity and AIRE mutation results. Out of autoimmune controls studied, two patients were positive for IFN-ω Ab. Positivity and levels of IFN-ω Ab in the APS-1 patients studied were similar irrespective of patient's clinical phenotype and AIRE genotype. Furthermore, IFN-ω Ab levels did not change over time (up to 36 years of disease duration) in 8 APS-1 patients studied. CONCLUSIONS: We have developed a novel, highly sensitive and specific assay for measurement of IFN-ω Ab. It provides a simple and convenient method for the assessment of patients with APS-1 and selecting patients suspected of having APS-1 for AIRE gene analysis.

Evaluation of the stiffness characteristics of rapid palatal expander screws.

BACKGROUND: The aim of this study is to evaluate the mechanical properties of the screws used for rapid expansion of the upper jaw. METHODS: Ten types of expansion screw were assessed, seven with four arms: Lancer Philosophy 1, Dentaurum Hyrax Click Medium, Forestadent Anatomic Expander type "S", Forestadent Anatomic Expander type "S" for narrow palates, Forestadent Memory, Leone A 2620-10 with telescopic guide, and Leone A 0630-10 with orthogonal arms; and three with two arms: Dentaurum Variety S.P., Target Baby REP Veltri, and Leone A 362113. A test expander with the mean dimensions taken from measurements on a sample of 100 expanders was constructed for each screw. The test expanders were connected to the supports of an Instron 4467 (Instron Corp., USA) mechanical testing machine equipped with a 500 N load cell, and the compression force exerted after each activation was measured. The mean forces expressed by the two- and four-arm expanders were then compared. RESULTS: After five activations, the forces expressed by the two-arm devices were double than those expressed by the four-arm devices on average (224 ± 59.9 N vs. 103 ± 32.9 N), and such values remained high after subsequent activations. CONCLUSIONS: The expanders tested demonstrated stiffness characteristics compatible with opening of the palatine sutures in pre-adolescent patients. The stiffness of such devices can be further increased during the construction phase.

Organ-specific autoimmunity in relation to clinical characteristics in children with long-lasting type 1 diabetes.

BACKGROUND: The aim of this study was to assess the prevalence of diabetes and other organ-specific autoantibodies (Ab) associated with various autoimmune conditions, in Polish children with type 1 diabetes mellitus (T1DM). METHODS: In this study 114 patients, aged 13.4 years, with mean diabetes duration 5.2 years were included. Ab to islet cell antigens: glutamic acid decarboxylase (GAD), insulinoma antigen 2 (IA-2), zinc transporter 8 (ZnT8), together with thyroid peroxidase Ab (TPO Ab), thyroglobulin Ab (Tg Ab), tissue transglutaminase Ab (tTG Ab) and 21-hydroxylase Ab (21-OH Ab) were measured. RESULTS: The prevalence of at least one diabetes associated Ab was found in 87%, with the highest prevalence of 64% for ZnT8 Ab. In patients with disease duration <5 years, at least one antibody was present in 90%, the most prevalent was ZnT8 Ab (72%). In patients with duration >10 years, 50% had at least one antibody. The prevalence of other than islet cell autoimmunity was high (34%). Thyroid Ab were detected in 26% patients, 42% in girls vs. 8% in boys, p<0.001. tTG Ab were found in 11% patients, with a greater prevalence in children with early onset (p=0.01). 21-OH Ab were found in 2.6% T1DM patients. CONCLUSIONS: Islet Ab were found in most T1DM children and remained positive even 10 years after onset. ZnT8 Ab emerged as an important marker for the diagnosis of T1DM in the Polish children. Screening for non-diabetes Ab in T1DM may be helpful in identifying subclinical cases of autoimmune thyroid, celiac or Addison's disease (AD).

Cranial reconstruction: 3D biomodel and custom-built implant created using additive manufacturing.

Additive manufacturing (AM) technology from engineering has helped to achieve several advances in the medical field, particularly as far as fabrication of implants is concerned. The use of AM has made it possible to carry out surgical planning and simulation using a three-dimensional physical model which accurately represents the patient's anatomy. AM technology enables the production of models and implants directly from a 3D virtual model, facilitating surgical procedures and reducing risks. Furthermore, AM has been used to produce implants designed for individual patients in areas of medicine such as craniomaxillofacial surgery, with optimal size, shape and mechanical properties. This work presents AM technologies which were applied to design and fabricate a biomodel and customized implant for the surgical reconstruction of a large cranial defect. A series of computed tomography data was obtained and software was used to extract the cranial geometry. The protocol presented was used to create an anatomic biomodel of the bone defect for surgical planning and, finally, the design and manufacture of the patient-specific implant.

AIRE gene mutations and autoantibodies to interferon omega in patients with chronic hypoparathyroidism without APECED.

OBJECTIVE: To assess autoimmune regulator (AIRE) gene mutations, class II HLA haplotypes, and organ- or non-organ-specific autoantibodies in patients with chronic hypoparathyroidism (CH) without associated Addison's disease (AD) or chronic candidiasis (CC). DESIGN, PATIENTS AND MEASUREMENTS: Twenty-four patients who had CH without AD or CC were included in the study. AIRE gene mutations in all 14 exons were studied using PCR in 24 patients, 105 healthy controls and 15 first-degree relatives of CH patients with AIRE mutations. Human leucocyte antigens (HLA) were determined for all 24 patients and 105 healthy controls. Autoantibodies to a range of antigens including NACHT leucine-rich-repeat protein-5 (NALP5) and interferon omega (IFNω) were tested in all 24 patients. RESULTS: AIRE gene mutations were found in 6 of 24 (25%) patients, all females, and this was significantly higher (P < 0·001) compared with AIRE mutations found in healthy controls (2/105). Three patients (12·5%) had homozygous AIRE mutations characteristic of Autoimmune-Poly-Endocrinopathy-Candidiasis-Ectodermal-Dystrophy and all three were also positive for IFNω-autoantibodies. Three patients (12·5%) had heterozygous AIRE mutations; two of these were novel mutations. One of the patients with heterozygous AIRE mutations was positive for both NACHT leucine-rich-repeat protein 5 and IFNω autoantibodies. Heterozygous AIRE mutations were found in 10 of 15 first-degree relatives of CH patients with AIRE mutations, although none was affected by CH. Class II HLA haplotypes were not statistically different in patients with CH compared to healthy controls. CONCLUSIONS: Analysis of AIRE gene mutations together with serum autoantibody profile should be helpful in the assessment of patients with CH, in particular young women with associated autoimmune diseases.