Investigating Trace Element Homeostasis In Vivo

Overview

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Key numbers

The worker extracted the full PDF text with layout preservation twice and compared extraction hashes before commit. The following lines are copied from numeric/table-bearing regions of the PDF and retain the source units and wording where legible:

Table 1: ICP-MS parameters for mercury quantification… 49
Table 2: Total content of the trace elements in feed and water in the animal study. … 50
Table 4: Thermal cycle conditions for cDNA synthesis. … 54
Table 5: Thermal cycle conditions for RT-qPCR. … 55
Table 6: Concentrations of total Hg in the basolateral compartment of the in vitro BBB
Table 7: Cellular Hg content in PBCECs and CCF-STTG1 cells grown on Transwell® filters after
Supplementary Table S2: Priming oligonucleotide sequences and RT-qPCR target
Supplementary Table S3: Overview of parameters determined in the animal experiment. … 114
Supplementary Table S4: Overview of statistical testing of determined parameters in
Supplementary Table S5: Comparison of the EC30 and IC50 values regarding different
Supplementary Table S6: List of chemicals… 124
demanding, requiring about 20% of the body’s total oxygen consumption (45), a disturbance of oxygen
sources, with 80 – 100% of total Hg in fish muscle and 50 – 80% of total Hg in seafood other than fish
(JECFA) has set a provisional tolerable weekly intake (PTWI) of 4 µg/kg body weight (bw) for inorganic
Hg (125) and a PTWI of 1.6 µg/kg bw for MeHg (126), respectively. These values are based on the most
for MeHg of 1.3 µg/kg bw (123), which indicates a more conservative approach to account for potential
Se uptake occurs mainly in the lower part of the small intestine with an efficiency of 70 to 95% (200).
promoting the formation of stable Hg-Se complexes (257, 260, 262). In addition, demethylation has
Cells were stored in a freezing medium composed of 90% fetal calf serum (FCS) and 10% dimethyl
reaching 80% confluence after two to three days. For passaging, cells were washed twice with
phosphate-buffered saline (PBS-UVC) and then treated with trypsin (0.25% (v/v) in PBS-EDTA) for cell
supplemented with 10% (v/v) FCS, 1.4 mM L-glutamine, 100 U/mL penicillin, and 100 mg/mL
passaging once a week, they were briefly washed with trypsin (0.25% (v/v) in PBS-EDTA) and then
placed in 70% (v/v) ice-cold ethanol and transported in ice-cold PBS containing 200 U/mL penicillin and
streptomycin, and 100 µg/mL gentamicin). Following a two-hour dispase digestion at 37 °C (6.5% (v/v)
(0.7 mM L-glutamine, 100/mL penicillin, 100 µg/mL streptomycin, 100 µg/mL gentamicin, and 10% FCS
glutamine, 100 U/mL penicillin, 100 µg/mL streptomycin, and 10% (v/v) FCS). The cells were trypsinised
(0.0625% (v/v) trypsin in PBS) before cryopreservation in FCS containing 10% DMSO, as described
were cultured in RPMI 1640, containing 10% (v/v) FCS, 1.4 mM L-glutamine, 100 U/mL penicillin, and
only results with a difference under 10% were included in the calculation of the mean.
medium before incubation for 3 h at 37 °C for dye uptake. The cells were washed with 100 µL of 0.5%
formaldehyde solution in PBS-UVC. Dye extraction was performed by adding 100 µL of an acidified (1%
were fixed with 3.7% formaldehyde in PBS-UVC for 10 min at 37°C and permeabilised with 2.2%
well plates. After incubation with the test compound, cells were fixed with 3.7% formaldehyde in PBS-
UVC for 30 min at RT. Cells were washed with a washing buffer (PBS-UVC containing 0.05% Tween®20)
following permeabilisation with 0.2% Triton™ X-100 in PBS-UVC for 15 min at RT. After another
washing step with washing buffer, the cells were blocked with 1% bovine serum albumin (BSA) in PBS-
trypsin (0.25% (v/v) in PBS-EDTA) for 3 min at 37 °C. The reaction was stopped by adding 300 µL of ice-
cold PBS-UVC with 5% (v/v) FCS. Cells were detached and transferred into 2 mL tubes, and kept on ice.
design of the spray chamber and sample line, only about 1-2% of the droplets enter the plasma.
demonstrated that using a preparation matrix containing 5% nitric acid (HNO3; v/v) and 5%
100 µg/L iridium (Ir); final concentration: 10 µg/L Ir) and 200 µL of an HNO3/HCl solution (50% (v/v)

Methods (brief)

QUANTIFICATION OF TOTAL MERCURY BY ICP-MS … 46
3.7.1 Sample Preparation – Acid Microwave-Assisted Digestion … 47
3.7.2 ICP-MS Measurement… 48
3.8.2 Element Quantification via ICP-MS/MS … 51
4.3.4 Analytical Considerations and Bias for Mercury Determination via ICP-MS … 79
A DILUTION SCHEME FOR ICP-MS SAMPLE PREPARATION … 109
Table 1: ICP-MS parameters for mercury quantification… 49
Table 3: ICP-MS/MS parameters for TE quantification. … 53
Supplementary Table S1: Dilution scheme of the samples from different time points and
experiments for microwave-assisted acid digestion and ICP-MS
ICP-MS Inductively coupled plasma mass spectrometry
ICP-MS/MS Tandem inductively coupled plasma mass spectrometry
LOD Limit of detection
LOQ Limit of quantification
The TE status in the cerebellum was determined by TE quantification via ICP-MS/MS analysis.
streptomycin, and 100 µg/mL gentamicin). Following a two-hour dispase digestion at 37 °C (6.5% (v/v)
nylon mesh (pore size: 180 µm). Capillaries were isolated by trituration before a second digestion to
consideration (1:5 for LUHMES and 1:2 for CCF-STTG1). Each sample was at least measured twice, and

Implications

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Update history

The five most recent substantive edits to this page, classified major (evidence or structure moved), correction (a published value or statement was wrong and has been fixed), or minor (narrative rewritten without changing the underlying evidence). Each description is derived from what the edit did to this page; the linked commit is the authoritative record, routine regeneration passes are excluded, and the full version history lives in git. When DOI minting comes online (see schema docs), each entry below will also link to a version-pinned DataCite DOI.

Commit	Date	Change	Description
b01ec52c	2026-08-04	major	2 sections added
d49e450f	2026-08-03	major	5 sections added; narrative text revised