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With the advance of precision medicine, the availability of tumor tissue for molecular analysis has become a limiting factor. This is particularly the case for bone metastases which are frequently occurring in cancer types such as prostate cancer. Due to the necessary decalcification process it was long thought that transcriptome analysis will not be feasible from decalcified formalin-fixed, paraffin-embedded (DFFPE) in a large manner. Here we demonstrate that mRNA extraction from DFFPE is feasible, quick, robust and reproducible and that decalcification does not hamper subsequent gene expression analysis. This might assist in implementing transcriptome analysis from DFFPE into every day practice.

Tripartite motif (TRIM) family proteins are post-translational protein modifiers with E3-ubiquitin ligase activity, thereby involved in various biological processes. The molecular mechanisms driving prostate cancer (PCa) bone metastasis (BM) are incompletely understood, and targetable genetic alterations are lacking in the majority of cases. Therefore, we aimed to explore the expression and potential functional relevance of 71 TRIM members in bone metastatic PCa. We performed transcriptome analysis of all human TRIM family members and 770 cancer-related genes in 29 localized PCa and 30 PCa BM using Nanostring. KEGG, STRING and Ubibrowser were used for further bioinformatic gene correlation and pathway enrichment analyses. Compared to localized tumors, six TRIMs are under-expressed while nine TRIMs are over-expressed in BM. The differentially expressed TRIM proteins are linked to TNF-, TGFβ-, PI3K/AKT- and HIF-1-signaling, and to features such as proteoglycans, platelet activation, adhesion and ECM-interaction based on correlation to cancer-related genes. The identification of TRIM-specific E3-ligase-substrates revealed insight into functional connections to oncogenes, tumor suppressors and cancer-related pathways including androgen receptor- and TGFβ signaling, cell cycle regulation and splicing. In summary, this is the first study that comprehensively and systematically characterizes the expression of all TRIM members in PCa BM. Our results describe post-translational protein modification as an important regulatory mechanism of oncogenes, tumor suppressors, and pathway molecules in PCa progression. Therefore, this study may provide evidence for novel therapeutic targets, in particular for the treatment or prevention of BM.

Ryanodine receptors (RyRs) are Ca2+-regulated Ca2+channels of 2.2-megadalton in muscles and neurons for calcium signaling. How Ca2+regulates ion conduction in the RyR channels remains elusive. We determined a 2.6-Å cryo-EM structure of rabbit skeletal muscle RyR1, and used multiscale dynamics simulations to elucidate cation interactions with RyR1. We investigated 21 potential cation-binding sites that may together rationalize biphasic Ca2+response of RyR1. The selectivity filter captures a cation hydration complex by hydrogen-bonding with both the inner and outer hydration shells of water molecules. Molecular dynamics simulations suggest that adjacent Ca2+ions moving in concert along ion-permeation pathway are separated by at least two cation-binding sites. Our analysis reveals that RyR1 has been evolved to favor its interactions with two hydration shells of cations.

Tetrabromobisphenol A (TBBPA) and cadmium chloride (CdCl2) are the typical representative pollutants of brominated flame retardants and heavy metals found in the air of e-waste recycling workshops. However, their metabolic kinetics through mixture inhalation is unknown. In the present study, 8-week old Institute of Cancer Research (ICR) male mice were whole-body exposed to TBBPA and CdCl2 mixtures by inhalation. Tissue samples were collected for TBBPA and cadmium (Cd) analysis at 2, 4, 6, and 8 weeks during exposure and at 4 and 8 weeks after the completion of the 8-week exposure period. TBBPA was mainly distributed to the lungs, liver, kidney, testis, and spleen, with a high amount accumulated in the brain, liver, and spleen. Cd was mainly distributed to the lungs, liver, and kidney, with a high amount accumulated in the liver, kidney, and testis and a low amount accumulated in brain and serum. Tissue burden of TBBPA and Cd in all organs increased in a dose- and time-dependent manner during the exposure period. However, 4 weeks after the completion of an 8-week exposure, TBBPA concentrations in the liver, testis, brain, and serum and Cd concentrations in the liver, testis, and kidney were higher than the corresponding tissue concentrations during the exposure period. The rapid accumulation of both TBBPA and Cd in the lungs after inhalation exposure indicated a high risk of the respiratory system diseases for workers in e-waste recycling workshops. In addition, the migration of both TBBPA and Cd from lungs to liver and testis may result in more complex toxic effects in vivo.

This study was designed to investigate a prevalent brominated flame retardant tetrabromobisphenol A (TBBPA) and four heavy metals of Pb, Cr, As, Cd in dust samples (52 indoor and 52 outdoor) collected from residential houses in an e-waste recycling area in Southeast China. For TBBPA, the mean concentration in indoor dust (3435 ng/g, dw) was higher than that in outdoor dust (1998 ng/g, dw). For heavy metals, the mean concentrations of Pb, Cr, As, Cd were 399, 151, 48.13, and 5.85 mg/kg in indoor dust, respectively, and were 328, 191, 17.59, and 4.07 mg/kg in outdoor dust, respectively. Except for As, concentrations of TBBPA and other metals decreased with the increased distance away from the e-waste recycling center, suggesting significant contribution of e-waste activities. The daily exposure doses of TBBPA ranged from 0.04 to 7.50 ng/kg-bw/day for adults and from 0.31 to 58.54 ng/kg-bw/day for children, representing the highest values reported to date for TBBPA exposure via dust ingestion. Daily exposure doses of Cr, As, and Cd were all below the reference doses. However, daily exposure dose of Pb for children in areas near the e-waste processing center was above the reference dose, posing significant health concern for children in that region.