Detection of SARS-CoV-2 infection in thyroid follicular cells from a COVID-19 autopsy series.

Objective: To understand whether thyroid cells can be directly infected by the SARS-CoV-2 virus and to establish a putative correlation with the expression of the host entry machinery: ACE-2, TMPRSS2, and furin. Methods: We assessed the presence of SARS-CoV-2 virus at the gene level by RT-PCR, viral RNA transcripts localization by in situ hybridization, and by detecting viral proteins by immunohistochemistry for the nucleocapsid and the spike proteins. Furthermore, we also described the immunoexpression of key host factors for virus entry in the COVID-19 thyroid samples. Results: We performed RT-PCR for SARS-CoV-2 in all autopsy specimens and detected viral genome positivity in 13 of 15 thyroid tissues and in a lung specimen. In 9 of the 14 positive samples, we were also able to confirm SARS-CoV-2 signal by in situ hybridization. Immunohistochemistry for the viral nucleocapsid and spike protein was also positive for ten and nine of the RT-PCR-positive cases, respectively, but revealed a lower sensitivity. We also described, for the first time in a COVID-19 series, the immunohistochemical expression of ACE-2, TMPRSS2, and furin in the thyroid. Conclusions: Our results obtained in thyroid specimens from deceased COVID-19 patients indicate that thyrocytes can be directly infected by SARS-CoV-2 since we detected the presence of SARS-CoV-2 genome in follicular cells. Nevertheless, we did not find a clear correlation between the presence of viral genome and the expression of the host factors for virus entry, namely ACE-2, TMPRSS2, and furin.

Searching for SARS-CoV-2 in Cancer Tissues: Results of an Extensive Methodologic Approach based on ACE2 and Furin Expression.

SARS-CoV-2 pandemics have been massively characterized on a global scale by the rapid generation of in-depth genomic information. The main entry gate of SARS-CoV-2 in human cells is the angiotensin-converting enzyme 2 (ACE2) receptor. The expression of this protein has been reported in several human tissues, suggesting a correlation between SARS-CoV-2 organotropism and ACE2 distribution. In this study, we selected (a series of) 90 patients who were submitted to surgery for tumor removal between the beginning of the SARS-CoV-2 pandemic and the closure of operating rooms (by the end of March 2020) in two different countries-Portugal and Brazil. We evaluated the expressions of ACE2 and furin (another important factor for virus internalization) in colon (n = 60), gastric (n = 19), and thyroid (n = 11) carcinomas. In a subseries of cases with PCR results for SARS-CoV-2 detection in the peri-operatory window (n = 18), we performed different methodological approaches for viral detections in patient tumor samples. Our results show that colon and gastric carcinomas display favorable microenvironments to SARS-CoV-2 tropism, presenting high expression levels of ACE2 and furin. From the subseries of 18 cases, 11 tested positive via PCR detection performed in tumor blocks; however, a direct association between the ACE2 expression and SARS-CoV-2 infection was not demonstrated in cancer cells using histology-based techniques, such as immunohistochemistry or in situ hybridization. This study raises the possibility of ACE2-mediated viral tropism in cancer tissues to be clarified in future studies.

Multi-element determination of metals and metalloids in waters and wastewaters, at trace concentration level, using electroanalytical stripping methods with environmentally friendly mercury free-electrodes: A review.

Nowadays, water is no longer regarded as an inexhaustible resource and the excessive release and proliferation of toxic metal(loid)s into aquatic environments has become a critical issue. Therefore, fast, accurate, simple, selective, sensitive and portable methodologies to detect multiple elements in natural waters is of paramount importance. Electrochemical stripping analysis is an efficient tool for trace metal(loid)s determinations and bring new prospects for answering the current environmental concerns. This review presents a survey of the advancements made between 2003 and 2016 on the development and application of non-toxic mercury free electrodes on the simultaneous analysis of metals and metalloids in waters and wastewaters by means of electroanalytical stripping techniques. The advantages, limitations, improvements and real applications of these "green" sensors are discussed from a critical point of view.

Determination of arsenic and antimony in seawater by voltammetric and chronopotentiometric stripping using a vibrated gold microwire electrode.

The oxidation potentials of As(0)/As(III) and Sb(0)/Sb(III) on the gold electrode are very close to each other due to their similar chemistry. Arsenic concentration in seawater is low (10-20 nM), Sb occurring at ~0.1 time that of As. Methods are shown here for the electroanalytical speciation of inorganic arsenic and inorganic antimony in seawater using a solid gold microwire electrode. Anodic stripping voltammetry (ASV) and chronopotentiometry (ASC) are used at pH ≤2 and pH 8, using a vibrating gold microwire electrode. Under vibrations, the diffusion layer size at a 5 µm diameter wire is 0.7 µm. The detection limits for the As(III) and Sb(III) are below 0.1 nM using 2 min and 10 min deposition times respectively. As(III) and Sb(III) can be determined in acidic conditions (after addition of hydrazine) or at neutral pH. In the latter case, oxidation of As(0) to As(III) was found to proceed through a transient As(III) species. Adsorption of this species on the gold electrode at potentials where Sb(III) diffused away is used for selective deposition of As(III). Addition of EDTA removes the interfering effect of manganese when analysing As(III). Imposition of a desorption step for Sb(III) analysis is required. Total inorganic arsenic (iAs=As(V)+As(III)) can be determined without interference from Sb nor mono-methyl arsenious acid (MMA) at 1.6

Simultaneous electrochemical determination of arsenic, copper, lead and mercury in unpolluted fresh waters using a vibrating gold microwire electrode.

In this work, a simple, rapid, reliable and low cost method for simultaneous electrochemical determination of As, Cu, Hg and Pb ions, on a vibrating gold microwire electrode combined with stripping voltammetry, is described for the first time. The multi-element detection was performed in the presence of oxygen by differential pulse anodic stripping voltammetry (DPASV) in HCl 0.1 M with NaCl 0.5 M. This media was found optimum in terms of peak resolution, peak shape and sensitivities, and has a composition similar to seawater to which the method could potentially be applied. The gold microwire electrode presented well defined, undistorted, sharp and reproducible peaks for trace concentrations of Cu, Hg and Pb and As presented a reproducible peak with a small shoulder. Using a gold vibrating microwire electrode of 25 µm diameter and 30s deposition time, the detection limits of As, Cu, Hg and Pb were 0.07, 0.4, 0.07 and 0.2 µgL(-1), respectively. Possible effects of Al, Cd, Cr, Fe, Mn, Ni, Sb and Zn were investigated but did not cause any significant interferences. Finally, the method was applied for the simultaneous determination of these four metals in unpolluted river water samples and the results were validated by Atomic Absorption Spectroscopy with Electrothermal Atomization (AAS-EA) or by Inductively Coupled Plasma Mass Spectrometry (ICP-MS).