Significance of Anti-TPO as an Early Predictive Marker in Thyroid Disease.

Even though most thyroid subjects are undiagnosed due to nonspecific symptoms, universal screening for thyroid disease is not recommended for the general population. In this study, our motive is to showcase the early appearance of thyroid autoantibody, anti-TPO, prior to the onset of thyroid hormone disruption; hence the addition of anti-TPO in conjunction with traditional thyroid markers TSH and FT4 would aid to reduce the long-term morbidity and associated health concerns. Here, a total of 4581 subjects were tested multiple times for TSH, FT4, anti-TPO, and anti-Tg and followed up for 2 years. We streamlined our subjects into two groups, A1 (euthyroid at first visit, but converted to subclinical/overt hypothyroidism in follow-up visits) and A2 (euthyroid at first visit, but converted to hyperthyroidism in follow-up visits). According to our results, 73% of hypothyroid subjects (from group A1) and 68.6% of hyperthyroid subjects (from group A2) had anti-TPO 252 (±33) and 277 (±151) days prior to the onset of the thyroid dysfunction, respectively. Both subclinical/overt hypothyroidism and hyperthyroidism showed a significantly higher percentage of subjects who had anti-TPO prior to the onset of thyroid dysfunction compared to the combined control group. However, there was no significant difference in the subjects who had anti-Tg earlier than the control group. Further assessment showed that only anti-TPO could be used as a standalone marker but not anti-Tg. Our results showcase that anti-TPO appear prior to the onset of thyroid hormone dysfunction; hence testing anti-TPO in conjunction with TSH would greatly aid to identify potentially risk individuals and prevent long-term morbidity.

Insights into cardiovascular risk and nutritional status in subjects with wheat-related disorders.

Objective: Wheat-related disorders are a spectrum of disorders associated with different autoimmune and non-autoimmune diseases. However, it is unclear whether these wheat-related disorders lead to adverse health effects such as cardiovascular risk, nutritional deficiencies etc. The objective of the study was to explore the lipid profiles and the nutritional status of subjects with wheat-related disorders to understand the potential threat of wheat on cardiovascular risk and nutritional deficiency. Method: A total of 1041 subjects who showed wheat-related symptoms were initially tested for the wheat protein antibody panel (Wheat Zoomer (WZ) panel and Coeliac Disease (CD) panel), then for cardiovascular panel and the micronutrient panel at Vibrant America Clinical Laboratory. Results: Subjects with both Wheat Zoomer positivity (WZ+) and Coeliac Disease positivity (CD+) had significantly low levels of high-density lipoproteins (HDL) (279/483(57.8%) and 29/47(61.7%) respectively), but only subjects with WZ + had low levels of Apo A1 (44/424(9.5%)), and high levels of Omega 6 fatty acids (53/334(15.9%)). None of the micronutrients tested showed a significant imbalance in WZ + subjects. Conclusion: Subjects with positive serology for WZ have deranged blood lipid profiles but did not show any significant micronutrient deficiency. Hence, our results showcase a significant association of wheat-related disorders to cardiovascular risk.

Analysis of Electrochemically Elusive Trace Metals with Carbon Fiber Microelectrodes.

There is great interest in rapidly monitoring metals of biological and environmental interest. Electrochemistry is traditionally a powerful tool for metal analysis but can be limited by its scope and low temporal resolution. The scope is limited by the potential window of the working electrode and rapid analysis is limited, in part, by the need for nucleation/growth for preconcentration. In prior work, we showed that a rapid equilibrium mediated preconcentration process facilitated fast scan cyclic voltammetry (FSCV) responses to Cu(II) and Pb(II) at carbon fiber microelectrodes (CFMs). In this manuscript, we apply this same principle to Ca(II), Al(III), Mg(II), and Zn(II), metal ions that are traditionally difficult to electroanalyze. We demonstrate FSCV reduction peaks for these four metals whose positions and amplitudes are dependent on scan rate. The adsorption profiles of these ions onto CFMs follow Langmuir's theory for monolayer coverage. We calculate the thermodynamic equilibrium constant of metal adsorption onto CFMs and find that these constants follow the same order as those previously reported by other groups on other activated carbon materials. Finally, a real-time complexation study is performed with ligands that have preference for divalent or multivalent ions to probe the selectivity of the real-time signal. We observe a linear relationship between formation constant ( kf) and % change in the FSCV signal and use this correlation to, for the first time, report the kf of an Al(III)-complex. This work demonstrates the versatility of FSCV as a method with capacity to extend the scope of rapid electroanalysis.

Exploring Systemic Autoimmunity in Thyroid Disease Subjects.

INTRODUCTION: Individuals with one autoimmune disease are at risk of developing a second autoimmune disease, but the pathogenesis or the sequential occurrence of multiple autoimmune diseases has not been established yet. In this study, we explored the association and sequential occurrence of antibodies in thyroid disease and systemic autoimmune disease subjects. We evaluated thyroid hormones, thyroid-stimulating hormone (TSH), free thyroxine (FT4), thyroid autoantibodies, anti-thyroperoxidase (anti-TPO), and anti-thyroglobulin (Tg) to comprehend the association with systemic autoimmune autoantibodies, anti-nuclear antibodies (ANA), and autoantibodies to extractable nuclear antigens (ENA) in subjects with thyroid-related symptoms. METHODS: A total of 14825 subjects with thyroid-related symptoms were tested at Vibrant America Clinical Laboratory for thyroid markers (TSH, FT4, anti-TPO, and anti-Tg) and an autoimmune panel (ANA panel and ENA-11 profile) from March 2016 to May 2018. Thyroid-positive (based on TSH and FT4 levels), anti-TPO-positive, and anti-Tg-positive subjects were assessed for the prevalence of ANA and anti-ENA antibodies. A 2-year follow-up study was conducted to assess the sequential order of appearance of autoimmune markers in thyroid and systemic autoimmune diseases. RESULTS: In the retrospective analysis, 343/1671 (20.5%), 2037/11235 (18.1%), and 1658/9349 (17.7%) of thyroid+, anti-TPO+, and anti-Tg+ subjects were found to be seropositive for ANA. Anti-ENA was detected in a higher prevalence than ANA with 475/1671 (28.4%), 3063/11235 (27.3%), and 2511/9349 (26.9%) in the same groups of subjects, respectively. Our results are found to be much higher than the reported prevalence of anti-ENA in general population. During the 2-year follow-up study, anti-TPO appeared significantly earlier than ANA and anti-ENA in an average of 253 (±139) and 227 (±127) days, respectively. CONCLUSIONS: A high prevalence of anti-ENA and ANA was found to be coexisting with autoimmune thyroid disease subjects, with anti-TPO occurring prior to the onset of ANA and anti-ENA. Therefore, frequent follow-ups and evaluation of ANA and anti-ENA in subjects with anti-TPO positivity would be beneficial in early detection of other systemic autoimmune diseases.

Fast voltammetry of metals at carbon-fiber microelectrodes: towards an online speciation sensor.

Speciation controls the chemical behavior of trace metals. Thus, there is great demand for rapid speciation analysis in a variety of fields. In this study, we describe the application of fast scan cyclic voltammetry (FSCV) and fast scan adsorption controlled voltammetry (FSCAV) to trace metal speciation analysis. We show that Cu2+ can be detected using FSCAV in different matrices. We find that matrices with different Cu2+ binding ability do not affect the equilibrium of Cu2+ adsorption onto CFMs, and thus are an excellent predictor for free Cu2+ ([Cu2+]free) in solution. We modelled a correlation between the FSCV response, [Cu2+]free and log Kf for 15 different Cu2+ complexes. Using our model, we rapidly predicted, and verified [Cu2+]free and Kf of a real groundwater sample spiked with Cu2+. We thus highlight the potential of fast voltammetry as a rapid trace metal speciation sensor.

Fast voltammetry of metals at carbon-fiber microelectrodes: rapid determination of solution formation constants.

Metal speciation controls the behavior of aqueous metal ions. Fundamental thermodynamic parameters, such as the formation constant (Kf) of metal-ligand equilibria, provide useful speciation information. Although this information can be determined by spectroscopic techniques with high accuracy, it comes at the expense of time and cost. In this work, we studied Cu2+ complexation with different ligands using an ultra-fast method, fast scan cyclic voltammetry (FSCV) at carbon-fiber microelectrodes (CFMs). We observed a correlation between the FSCV response and the previously reported Cu2+-ligand equilibrium constants. This relationship allowed us to model a predictive relationship between Kf and 16 model ligands. We hence present an essential proof of principle study that highlights FSCV's capability to prove speciation information in real time.

Voltammetric Characterization of Cu(II) Complexation in Real-Time.

Aqueous metal behavior is strongly regulated by speciation, which in turn is highly dependent on complexation. Trace metal complexation is difficult to characterize in dynamically changing systems due to a lack of analytical methods that can rapidly report free-metal concentrations. In this paper, we perform proof-of-principle experiments that demonstrate the utility of fast-scan cyclic voltammetry (FSCV) for providing speciation information in real-time by characterizing dynamic Cu(II) binding. We study Cu(II) FSCV responses in 3-(N-morpholino)propanesulfonic acid (MOPS) buffer and characterize the hydrodynamic aspects of our experimental setup (continuously stirred tank reactor). We observe Cu(II) complexation in real-time using five ligands with differing formation constants of Cu(II) complexation. Finally, we utilize geochemical models to fit our real-time experimental Cu(II)-binding curves. Our proof-of-principle experiments show that FSCV is a powerful tool for studying real-time Cu(II) complexation, which is essential speciation information for better interpretation of Cu(II) behavior in dynamically changing systems, such as those encountered in biology or the environment.

Real-time subsecond voltammetric analysis of Pb in aqueous environmental samples.

Lead (Pb) pollution is an important environmental and public health concern. Rapid Pb transport during stormwater runoff significantly impairs surface water quality. The ability to characterize and model Pb transport during these events is critical to mitigating its impact on the environment. However, Pb analysis is limited by the lack of analytical methods that can afford rapid, sensitive measurements in situ. While electrochemical methods have previously shown promise for rapid Pb analysis, they are currently limited in two ways. First, because of Pb's limited solubility, test solutions that are representative of environmental systems are not typically employed in laboratory characterizations. Second, concerns about traditional Hg electrode toxicity, stability, and low temporal resolution have dampened opportunities for in situ analyses with traditional electrochemical methods. In this paper, we describe two novel methodological advances that bypass these limitations. Using geochemical models, we first create an environmentally relevant test solution that can be used for electrochemical method development and characterization. Second, we develop a fast-scan cyclic voltammetry (FSCV) method for Pb detection on Hg-free carbon fiber microelectrodes. We assess the method's sensitivity and stability, taking into account Pb speciation, and utilize it to characterize rapid Pb fluctuations in real environmental samples. We thus present a novel real-time electrochemical tool for Pb analysis in both model and authentic environmental solutions.