Lucia Terzuoli

Overcoming a "Probable" diagnosis in antimitochondrial antibody negative primary biliary cirrhosis: Study of 100 sera and review of the literature

Serum anti-mitochondrial antibodies (AMA) are the serological hallmark of primary biliary cirrhosis (PBC), yet up to 15% of PBC sera are AMA negative at routine indirect immunofluorescence (IIF) while being referred to as “probable” cases. The diagnostic role of PBC-specific antinuclear antibodies (ANA) remains to be determined. We will report herein data on the accuracy of new laboratory tools for AMA and PBC-specific ANA in a large series of PBC sera that were AMA-negative at IIF. We will also provide a discussion of the history and current status of AMA detection methods. We included IIF AMA-negative PBC sera (n = 100) and sera from patients with other chronic liver diseases (n = 104) that had been independently tested for IIF AMA and ANA; sera were blindly tested with an ELISA PBC screening test including two ANA (gp210, sp100) and a triple (pMIT3) AMA recombinant antigens. Among IIF AMA-negative sera, 43/100 (43%) manifested reactivity using the PBC screening test. The same test was positive for 6/104 (5.8%) control sera. IIF AMA-negative/PBC screen-positive sera reacted against pMIT3 (11/43), gp210 (8/43), Sp100 (17/43), both pMIT3 and gp210 (1/43), or both pMIT3 and Sp100 (6/43). Concordance rates between the ANA pattern on HEp-2 cells and specific Sp100 and gp210 ELISA results in AMA-negative subjects were 92% for nuclear dots and Sp100 and 99% for nuclear rim and gp210. Our data confirm the hypothesis that a substantial part of IIF AMA-negative (formerly coined “probable”) PBC cases manifest disease-specific autoantibodies when tested using newly available tools and thus overcome the previously suggested diagnostic classification. As suggested by the recent literature, we are convinced that the proportion of AMA-negative PBC cases will be significantly minimized by the use of new laboratory methods and recombinant antigens.

Comparative determination of purine compounds in carotid plaque by capillary zone electrophoresis and high-performance liquid chromatography

Allantoin, uric acid (UA), hypoxanthine (Hx) and xanthine (X) were determined on carotid plaque by capillary zone electrophoresis (CZE) and high-performance liquid chromatography (HPLC). Comparison of the results showed that capillary zone electrophoresis may have similar or even superior analytical performance to HPLC, especially for the determination of allantoin in biological samples.

Association between stressful life events and autoimmune diseases: A systematic review and meta-analysis of retrospective case–control studies

BACKGROUND Evidence of a relationship between stressful life events and the onset of autoimmune diseases is not univocal and there are no meta-analyses in the literature on the question. AIM To look for differences in the number and type of stressful life events in the premorbid period between patients with autoimmune diseases and healthy subjects. METHOD Review of the literature in PubMed and Scopus (January 1963-May 2015). INCLUSION CRITERIA We included retrospective case-control studies that compared patients diagnosed with autoimmune disorders and controls regarding the incidence of stressful events occurring before diagnosis, and investigated said events with validated questionnaires. EFFECT-SIZE INDEXES By random effect meta-analysis, two independent researchers calculated effect-size indexes as the difference between the means of the clinical groups and the control group in relation to the combined standard deviation. RESULTS The database searches produced 2490 articles, 14 of which were selected (3201 patients). Analysis showed a moderate but significant mean effect-size index [d=0.63, p<0.01], suggesting that autoimmune disorders are effectively associated with major stressful events in the premorbid period. The relationship between stressful events and autoimmune disease was weaker in studies with a high proportion of female subjects [β=-0.004, p<0.01] and stronger in studies that considered a longer interval between stressors and onset of disease [β=0.16, p<0.01]. CONCLUSIONS The results of this meta-analysis suggest that stressors may play an important role in the etiopathogenesis of autoimmune disorders. Only prospective studies can provide more certain inference about the causality of this relationship.

Fatty acid composition of phospholipids, triglycerides and cholesterol in serum of castrated and estradiol treated rats.

We have studied the levels of phospholipids, triglycerides, cholesterol esters, and their fatty acid composition in serum for normal, castrated and estradiol treated rats. The sex hormones did not greatly affect the levels of the various lipid fractions which did not undergo great significant variations, under the different treatments. More evident variations occurred in the percent composition of fatty acid and in the content of the various saturated (SAT), unsaturated (UNSAT), essential (EFA) and non essential fatty acids (NEFA). We studied the most important ratios: EFA/NEFA; UNS/SAT; 16:0/16:1; 18:0/18:1, 18:2/18:3; 18:2/20:4. 16:0/16:1; 18:0/18:1 represent the delta9 desaturase, one specific for palmitic, the other for stearic acid. 18:2/18:3 ratio is an index of the delta6 desaturase activity: 18:2/20:4 ratio of delta5 desaturase-elongase. Most changes were evident in triglycerides. We observed a different behaviour of the UNS/SAT and EFA/NEFA ratios in phospholipids and cholesterol esters, which may reflect either an effect of the sex hormones on the exchange of fatty acids between the same lipid fractions, or a redistribution of lipids among different tissues. Great variations were observed of the ratios 16:0/16:1; 18:0/18:1; 18:2/18:3; 18:2/20:4, which are ascribed a different effect of the sex hormones of delta9, delta6, delta5 desaturases.

Determination and separation of allantoin, uric acid, hypoxanthine, and xanthine by capillary zone electrophoresis.

Hypoxanthine, xanthine, uric acid and allantoin are the main products of purine nucleotide catabolism and they are formed through the sequence: nucleotide→ hypoxanthine → xanthine→ uric acid→ allantoin. Under normal conditions, they represent the balance between synthesis and breakdown of nucleotides. Their levels change, for example, under oxidizing conditions and may be useful for quantifying oxidant generation in human extracts. Uric acid is oxidized to allantoin by variuos reactive oxygen species and is thought to act as an antioxidant in human bodily fluids. Allantoin concentrations may therefore indicate free radical damage in vivo.

Celiac and non-celiac gluten sensitivity: a review on the association with schizophrenia and mood disorders

An association between many psychiatric and gluten-related disorders has been known for some time. In the case of schizophrenia and mood disorders, the major psychiatric disorders, there is much evidence, not without contradictions, of a possible association between schizophrenia and celiac disease. The association between mood disorders and gluten-related disorders, especially celiac disease, has only been studied for depression, often coupled with anxiety, and very recently for bipolar disorder. Since non-celiac gluten sensitivity is now known to be different from celiac disease, many studies have shown that gluten sensitivity is also associated with major psychiatric disorders. Here we review the literature on the association between schizophrenia/mood disorders and celiac disease/gluten sensitivity, pointing out the differences between these associations.

Assessment of a Test for the Screening and Diagnosis of Celiac Disease

Celiac disease (CD) is an immune‐mediated intolerance to dietary gluten, affecting genetically predisposed individuals. ELISA‐based serological tests help to decide if further duodenal biopsy is necessary, for this the diagnostic kits have to be accurate, specific, and sensible. In this study, we investigate the performance of an ELISA that uses the purified cross‐linked complex of tissue transglutaminase and gliadin, referred as the “neoepitope” (AESKULISA® tTG New Generation), as antigen.

Identification and structural characterization of a transient radical species in the uricase reaction mechanism

Uricase catalyzes the oxidation of urate to form allantoin, carbon dioxide and hydrogen peroxide. In this article, we demonstrate for the first time the presence of a radical intermediate involved in the reaction mechanism. Such radical species was entrapped using 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide as spin trap and the relative adduct was detected by electron paramagnetic resonance (EPR) technique. A structure of such radical (5-hydroperoxy isourate) is proposed, through chemical results and density functional theory calculations of the EPR coupling constants.

Decreased plasma endogenous soluble RAGE, and enhanced adipokine secretion, oxidative stress and platelet/coagulative activation identify non-alcoholic fatty liver disease among patients with familial combined hyperlipidemia and/or metabolic syndrome

OBJECTIVE In patients with familial combined hyperlipidemia (FCHL), without metabolic syndrome (MS), occurrence of non-alcoholic fatty liver disease (NAFLD) is related to a specific pro-inflammatory profile, influenced by genetic traits, involved in oxidative stress and adipokine secretion. Among FCHL or MS patients, hyperactivity of the ligand-receptor for advanced glycation-end-products (RAGE) pathway, as reflected by inadequate protective response by the endogenous secretory (es)RAGE, in concert with genetic predisposition, may identify those with NAFLD even before and regardless of MS. METHODS We cross-sectionally compared 60 patients with vs. 50 without NAFLD. Each group included patients with FCHL alone, MS alone, and FCHL plus MS. RESULTS NAFLD patients had significantly lower plasma esRAGE, IL-10 and adiponectin, and higher CD40 ligand, endogenous thrombin potential and oxidized LDL. The effects of MS plus FCHL were additive. The genotypic cluster including LOX-1 IVS4-14A plus ADIPO 45GG and 256 GT/GG plus IL-10 10-1082G, together with higher esRAGE levels highly discriminate FCHL and MS patients not developing NAFLD. CONCLUSIONS Among FCHL or MS patients, noncarriers of the protective genotypic cluster, with lower esRAGE and higher degree of atherothrombotic abnormalities coincide with the diagnosis of NAFLD. This suggests an interplay between genotype, adipokine secretion, oxidative stress and platelet/coagulative activation, accelerating NAFLD occurrence as a proxy for cardiovascular disease.

Detection of Autoantibodies Against Actin Filaments in Celiac Disease

Serum autoantibodies specifically directed toward intracellular cytoskeletal actin filaments (anti‐actin antibodies, AAA) were found to be associated with intestinal villous atrophy (IVA) in celiac disease (CD). The aim of this study was to assess IgA‐AAA with a commercial test that uses sections of rat intestinal epithelial cells in a well‐selected cohort of patients and to evaluate the relationship between the presence of serum IgA‐AAA and the severity of intestinal mucosa damage. J. Clin. Lab. Anal. 27:21–26, 2013.