Multi-element analysis of metals in human pathological and unchanged thyroid glands - pilot study.

Disturbances in the homeostasis of the elemental composition of thyroid tissue may have serious metabolic and health consequences. It is believed that the accumulation of some metals or the deficiency of others may even cause lethal tumours. Due to the fact that metallomics most often uses human serum to analyse macro and microelements as well as trace elements, it was decided to use material that is more difficult to obtain, but also adds credibility to the research - thyroid tissue samples biopsy. The experiments were conducted on 17 patients diagnosed with: nodular (10) and colloidal goitre (2), chronic thyroiditis (2), follicular adenoma (2) and papillary carcinoma (1). They were recruited by collecting a tumour fragment, control fragment and serum from each of them. The content of Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Zn was examined using ICP-OES (Inductively Coupled Plasma - Optical Emission Spectrometers). Simultaneously, biochemical methods were used to determine the markers of inflammation, glycation and peroxidation: malondialdehyde, pentosidine, reactive free amine content, compounds with thiol groups and galectin 3 in the sera of the examined patients. Three statistically significant correlations were identified: Ca-Mg and Cu-Zn in control tissues (p < 0.05) and Cr-Mn in pathological tissues (p < 0.05). A comparison of individual groups of patients shows that there are some potentail tendencies to increase or decrease in the concentration of certain elements or markers of inflammation and glycation, therefore we discuss potential relationships between a given parameter and a thyroid disorder. The pilot study is an introduction to a deeper analysis aimed at tracing the pathomechanism of the development of thyroid diseases, so that the risk of developing these diseases can be effectively minimized.

Association between Leukocyte Cell-Derived Chemotaxin 2 and Metabolic and Renal Diseases in a Geriatric Population: A Pilot Study.

LECT2 is not a routine diagnostic marker for any disease, but it has been associated with many pathologies, including systemic amyloidosis, rheumatoid arthritis, diabetes, atherosclerosis, and metabolic syndrome. With human aortic sections (n = 22) and sera from geriatric subjects (n = 79), we analyzed the relationships that could be observed between this protein and other parameters related to metabolic diseases. As a result, we observed a relatively high (r~0.8, p < 0.05) positive correlation between SRA and LECT2 and a negative correlation between EGFR and LECT2 (r~-0.4, p < 0.05). We observed LECT2 expression in macrophages, myocytes, and other aortic cells, with a tendency to be overexpressed in developed atherosclerotic plaques. We conclude that LECT2 exerts its chemotactic effects not only as a protein synthesized in the liver and secreted and circulating in the blood but also as a locally expressed protein within atherosclerotic plaque development. The LECT2-EGFR correlation suggests an association of this protein with loss of normal renal function. This fact can be associated with LECT2 amyloidosis, although it should be verified whether in the geriatric population there is indeed a widespread accumulation of LECT2 with the progression of aging or whether it is rather a marker of general deterioration of renal function.

How Diabetes and Other Comorbidities of Elderly Patients and Their Treatment Influence Levels of Glycation Products.

Medical care for geriatric patients is a great challenge, mainly due to various overlapping deficits relevant to numerous coexisting diseases, of which the most common are diabetes mellitus and atherosclerosis. In the case of diabetes, the glycation process is intensified, which accelerates atherosclerosis development and diabetic complications. Our goal was to investigate the relationship between the classical biochemical parameters of diabetes and atherosclerosis, as well as parameters which may indicate a nephropathy, and the parameters strictly related to glycation, taking into account the pharmacological treatment of patients. Methods: We analyzed the patients' serum concentrations of fluorescent glycation product-pentosidine, concentrations of soluble receptors for advanced glycation products (sRAGE), lipoprotein receptor-1 (LOX-1), galectin 3 (GAL3), scavenger receptor class A (SR-A), and scavenger receptor class B (SR-BI), as well as the level of lipid peroxidation and free amine content. Among the identified correlations, the most interesting are the following: sRAGE with triglycerides (r = 0.47, p = 0.009), sRAGE with SR-BI (r = 0.47, p = 0.013), SR-BI with LOX-1 (r = 0.31, p = 0.013), and SR-BI with HDL (r = -0.30, p = 0.02). It has been shown that pentosidine and reactive free amine contents are significantly higher in elderly patients with ischemic heart disease. Pentosidine is also significantly higher in patients with arterial hypertension. Malondialdehyde turned out to be higher in patients with diabetes mellitus type 2 that was not treated with insulin or metformin than in those treated with both medications (p = 0.052). GAL3 was found to be lower both in persons without diabetes and in diabetics treated with metformin (p = 0.005). LOX-1 was higher in diabetic patients not treated with metformin or insulin, and lowest in diabetics treated with both insulin and metformin, with the effect of metformin reducing LOX-1 levels (p = 0.039). Our results were the basis for a discussion about the diagnostic value in the clinical practice of LOX-1 and GAL3 in geriatric patients with diabetes and also provide grounds for inferring the therapeutic benefits of insulin and metformin treatment.

Contribution of Glycation and Oxidative Stress to Thyroid Gland Pathology-A Pilot Study.

The patho-mechanism of changes in the thyroid gland, including carcinogenesis, is a complex process, which involves oxidative stress. The goal of our investigation was to verify the extent of stress in the thyroid gland related to glycation. The study samples were comprised of blood sera, thyroid, and adipose tissue sections probed from 37 patients diagnosed with thyroid cancers and goiter. Using immuno-enzymatic and fluorometric assays we analyzed the content of advanced glycation end-products (AGEs), pentosidine, receptors for advanced glycation end-products (RAGE), scavenger receptor class (SR)-A, SR-B, glutathione, malondialdehyde and nitric oxide synthase. In addition to classic AGEs, a recent study detected the melibiose-derived glycation (MAGE) product. We demonstrated the presence of AGEs, MAGE and their receptors of the RAGE and SR-A. In addition, in the control samples of thyroid glands SR-B groups were detected as well as of pathological groups without noticeable tendency to antigen concentration in the area of carcinogenesis. Fluorescent AGEs correlate positively with glutathione, which supports the assumption that glycation stress leads to augmentation of oxidative stress and increase of the intensity of antioxidant mechanisms.

The diagnostic usefulness of the basophil activation test (BAT) with annexin V in an allergy to Alternaria alternata.

BACKGROUND: The basophil activation test (BAT) is an effective diagnostic tool in mold allergy, which is still not sufficiently known. OBJECTIVES: The aim of our study was to assess the degree of annexin V binding to the surface of the basophil cell membrane after stimulation with anti-immunoglobulin E (anti-IgE) and Alternaria alternata allergenic extract. MATERIAL AND METHODS: Alternaria alternata allergic patients (n = 32) and healthy volunteers (n = 33) were evaluated using skin prick tests (SPT), quantification of specific IgE (sIgE) and the BAT. Basophil activation was detected as a percentage degree of annexin V binding to the surface of the basophil cell membrane. RESULTS: Receiver operating characteristic (ROC) curve analysis yielded a threshold value of 4.95% of activated basophils when the tested group and control group were studied, with a sensitivity and specificity of 100% (area under curve [AUC] = 1; p = 0.00000) for 100 SBU/mL Alternaria alternata allergen extract. The threshold value was 10.28% with a sensitivity of 93.8% and specificity of 100% (AUC = 0.98958; p = 0.00000) for 10 SBU/mL mold extract, and 9.37% with a sensitivity of 90.3% and specificity of 100% (AUC = 0.96307; p = 0.00000) for 1 SBU/mL Alternaria alternata allergen extract. The method was least efficacious in antiIgE stimulation, where the threshold value was 5.48% with a sensitivity of 90.6% and specificity of 30.3% (AUC = 0.46780; p = 0.67039). CONCLUSIONS: The BAT with annexin V and sIgE measurement against Alternaria alternata increase the capability of a diagnostic laboratory for detecting mold sensitization. Both methods may certainly replace SPT, which are currently routinely used in allergy diagnosis. Annexin V may be considered a new basophil activation marker with an efficacy comparable to that of CD63 or CD203c.

Genome-wide analysis of gene expression after one year of venom immunotherapy.

Background Insect venom immunotherapy (VIT) is used to protect patients against Hymenoptera insects' venom allergy (HVA), which can result in severe systemic or even life-threatening conditions. Molecular mechanisms triggered by VIT remain largely unknown. Objective To compare genome-wide gene expression of patients with severe HVA prior to VIT and 12 months after. Methods Whole blood RNA samples were analyzed on an expression array. Results from differential expression obtained on a microarray platform were confirmed by quantitative real -time PCR (qRT-PCR). Subsequently we applied unsupervised clustering. Relative blood cell proportions and gene expression profiles were used as an input to csSAM to compute cell specific differential gene expression. Finally, transcription factor enrichment analysis was performed in MotifLab. Results & conclusions Comparison of genome-wide expression patterns for whole blood and qRT-PCR experiments revealed no significantly up and/or down regulated genes. This has been corroborated by unsupervised clustering. We found a significant upregulation of 26 genes in macrophages, of 15 genes in monocytes and 2 genes in T regulatory cells (Tregs). Analysis of the promoter sequences of these upregulated genes revealed a significant over-representation of binding motifs specific for kruppel-like factor 4, retinoic acid receptor gamma, and vitamin D receptor. Our results indicate that changes of gene expression invoked by VIT in peripheral blood may have a too small effect to be detected by conventional biostatistical approaches. When blood cell composition was taken into account we uncovered numerous changes of cell-specific gene expression. Given the regulatory implications we hypothesize that above-mentioned alterations may contribute to activation of anti-inflammatory signals in the innate branch of the immune system.