Green steel from red mud through climate-neutral hydrogen plasma reduction.

Red mud is the waste of bauxite refinement into alumina, the feedstock for aluminium production1. With about 180 million tonnes produced per year1, red mud has amassed to one of the largest environmentally hazardous waste products, with the staggering amount of 4 billion tonnes accumulated on a global scale1. Here we present how this red mud can be turned into valuable and sustainable feedstock for ironmaking using fossil-free hydrogen-plasma-based reduction, thus mitigating a part of the steel-related carbon dioxide emissions by making it available for the production of several hundred million tonnes of green steel. The process proceeds through rapid liquid-state reduction, chemical partitioning, as well as density-driven and viscosity-driven separation between metal and oxides. We show the underlying chemical reactions, pH-neutralization processes and phase transformations during this surprisingly simple and fast reduction method. The approach establishes a sustainable toxic-waste treatment from aluminium production through using red mud as feedstock to mitigate greenhouse gas emissions from steelmaking.

The high optical brightness of the BlueWalker 3 satellite.

Large constellations of bright artificial satellites in low Earth orbit pose significant challenges to ground-based astronomy1. Current orbiting constellation satellites have brightnesses between apparent magnitudes 4 and 6, whereas in the near-infrared Ks band, they can reach magnitude 2 (ref. 2). Satellite operators, astronomers and other users of the night sky are working on brightness mitigation strategies3,4. Radio emissions induce further potential risk to ground-based radio telescopes that also need to be evaluated. Here we report the outcome of an international optical observation campaign of a prototype constellation satellite, AST SpaceMobile's BlueWalker 3. BlueWalker 3 features a 64.3 m2 phased-array antenna as well as a launch vehicle adaptor (LVA)5. The peak brightness of the satellite reached an apparent magnitude of 0.4. This made the new satellite one of the brightest objects in the night sky. Additionally, the LVA reached an apparent V-band magnitude of 5.5, four times brighter than the current International Astronomical Union recommendation of magnitude 7 (refs. 3,6); it jettisoned on 10 November 2022 (Universal Time), and its orbital ephemeris was not publicly released until 4 days later. The expected build-out of constellations with hundreds of thousands of new bright objects1 will make active satellite tracking and avoidance strategies a necessity for ground-based telescopes.

2,6-Bis(5-(tert-butyl)-1H-pyrazol-3-yl)pyridine: Effects of the Peripheral Aliphatic Side Chain on the Coordination of Actinides(III) and Lanthanides(III).

To improve our understanding of the interaction mechanism in trivalent lanthanide and actinide complexes, studies with structurally different hard and soft donor ligands are of great interest. For that reason, the coordination chemistry of An(III) and Ln(III) with 2,6-bis(5-(tert-butyl)-1H-pyrazol-3-yl)pyridine (C4-BPP) has been explored. Time-resolved laser fluorescence spectroscopy (TRLFS) studies have revealed the formation of [Cm(C4-BPP)n]3+ (n = 1-3) (log ß1' = 7.2 ± 0.4, log ß2' = 10.1 ± 0.5, and log ß3' = 11.8 ± 0.6) and [Eu(C4-BPP)m]3+ (m = 1-2) (log ß1' = 4.9 ± 0.2 and log ß2' = 8.0 ± 0.4). The absence of the [Eu(C4-BPP)3]3+ complex shows a more favorable complexation of Cm(III) over that of Eu(III). Additionally, complementary NMR measurements have been conducted to examine the M(III)-N bond in Ln(III) and Am(III) C4-BPP complexes. 15N NMR data have revealed notable differences in the chemical shifts of the coordinating nitrogen atoms between the Am(III) and Ln(III) complexes. In the Am(III) complex, the coordinating nitrogen atoms have shown a shift by 260 ppm, indicating a higher fraction of covalent bonding in the Am(III)-N bond compared with the Ln(III)-N bond. This observation aligns excellently with the differences in the stability constants obtained from TRLFS studies.

Insights into the Complexation Mechanism of a Promising Lipophilic PyTri Ligand for Actinide Partitioning from Spent Nuclear Fuel.

The challenging issue of spent nuclear fuel (SNF) management is being tackled by developing advanced technologies that point to reduce environmental footprint, long-term radiotoxicity, volumes and residual heat of the final waste, and to increase the proliferation resistance. The advanced recycling strategy provides several promising processes for a safer reprocessing of SNF. Advanced hydrometallurgical processes can extract minor actinides directly from Plutonium and Uranium Reduction Extraction raffinate by using selective hydrophilic and lipophilic ligands. This research is focused on a recently developed N-heterocyclic selective lipophilic ligand for actinides separation to be exploited in advanced Selective ActiNide EXtraction (SANEX)-like processes: 2,6-bis(1-(2-ethylhexyl)-1H-1,2,3-triazol-4-yl)pyridine (PyTri-Ethyl-Hexyl-PTEH). The formation and stability of metal-ligand complexes have been investigated by different techniques. Preliminary studies carried out by electrospray ionization mass spectrometry (ESI-MS) analysis enabled to qualitatively explore the PTEH complexes with La(III) and Eu(III) ions as representatives of lanthanides. Time-resolved laser fluorescence spectroscopy (TRLFS) experiments have been carried out to determine the ligand stability constants with Cm(III) and Eu(III) and to better investigate the ligand complexes involved in the extraction process. The contribution of a 1:3 M/L complex, barely identified by ESI-MS analyses, was confirmed as the dominant species by TRLFS experiments. To shed light on ligand selectivity toward actinides over lanthanides, NMR investigations have been performed on PTEH complexes with Lu(III) and Am(III) ions, thereby showing significant differences in chemical shifts of the coordinating nitrogen atoms providing proof of a different bond nature between actinides and lanthanides. These scientific achievements encourage consideration of this PyTri ligand for a potential large-scale implementation.

Phosphorus solubility and dynamics in a tropical soil under sources derived from wastewater and sewage sludge.

Conventional phosphate fertilizers are usually highly water-soluble and rapidly solubilize when moistened by the soil solution. However, if this solubilization is not in alignment with plants demand, P can react with the soil colloidal phase, becoming less available over time. This is more pronounced in acidic, oxidic tropical soils, with high P adsorption capacity, reducing the efficiency of P fertilization. Furthermore, these fertilizers are derived from phosphate rock, a non-renewable resource, generating an environmental impact. To assess these concerns, waste-recycled P sources (struvite, hazenite and AshDec®) were studied for their potential of reducing P fixation by the soil and improving the agronomic efficiency of the P fertilization. In our work, we compared the solubilization dynamics of struvite, hazenite, AshDec® to triple superphosphate (TSP) in a sandy clay loam Ferralsol, as well as their effect on solution pH and on soil P pools (labile, moderately-labile and non-labile) via an incubation experiment. Leaching columns containing 50 g of soil with surface application of 100 mg per column (mg col-1) of P from each selected fertilizer and one control (nil-P) were evaluated for 60 days. Daily leachate samples from the column were analyzed for P content and pH. Soil was stratified in the end and submitted to P fractionation. All results were analyzed considering p < 0.05. Our findings showed that TSP and struvite promoted an acid P release reaction (reaching pHs of 4.3 and 5.5 respectively), while AshDec® and hazenite reaction was alkaline (reaching pHs of 8.4 and 8.5 respectively). Furthermore, TSP promoted the highest P release among all sources in 60 days (52.8 mg col-1) and showed rapid release dynamic in the beginning, while struvite and hazenite showed late release dynamics and lower total leached P (29.7 and 15.5 mg col-1 P respectively). In contrast, no P-release was detected in the leachate of the AshDec® over the whole trial period. Struvite promoted the highest soil labile P concentration (7938 mg kg-1), followed by hazenite (5877 mg kg-1) and AshDec® (4468 mg kg-1), all higher than TSP (3821 mg kg-1), while AshDec® showed high moderately-labile P (9214 mg kg-1), reaffirming its delayed release potential.

5-Aminosalicylic acid inhibits stem cell function in human adenoma-derived cells: implications for chemoprophylaxis in colorectal tumorigenesis.

BACKGROUND: Most colorectal cancers (CRC) arise sporadically from precursor lesions: colonic polyps. Polyp resection prevents progression to CRC. Risk of future polyps is proportional to the number and size of polyps detected at screening, allowing identification of high-risk individuals who may benefit from effective chemoprophylaxis. We aimed to investigate the potential of 5-aminosalicylic acid (5-ASA), a medication used in the treatment of ulcerative colitis, as a possible preventative agent for sporadic CRC. METHODS: Human colorectal adenoma (PC/AA/C1, S/AN/C1 and S/RG/C2), transformed adenoma PC/AA/C1/SB10 and carcinoma cell lines (LS174T and SW620) were treated with 5-ASA. The effect on growth in two- and three-dimensional (3D) culture, ß-catenin transcriptional activity and on cancer stemness properties of the cells were investigated. RESULTS: 5-ASA was shown, in vitro, to inhibit the growth of adenoma cells and suppress ß-catenin transcriptional activity. Downregulation of ß-catenin was found to repress expression of stem cell marker LGR5 (leucine-rich G protein-coupled receptor-5) and functionally suppress stemness in human adenoma and carcinoma cells using 3D models of tumorigenesis. CONCLUSIONS: 5-ASA can suppress the cancer stem phenotype in adenoma-derived cells. Affordable and well-tolerated, 5-ASA is an outstanding candidate as a chemoprophylactic medication to reduce the risk of colorectal polyps and CRC in those at high risk.

Impact of Solvent Polarity on the Ligand Configuration in Tetravalent Thorium N-Donor Complexes.

A combined NMR spectroscopic and theoretical study on the complexation of diamagnetic Th(IV) with 2,6-bis(5,6-dipropyl-1,2,4-triazin-3-yl)pyridine (nPr-BTP) was performed. Different ligand configurations were observed for [Th(nPr-BTP)3]4+ complexes depending on the solvent's ability to actively form hydrogen bonds. In polar aprotic solvents, a complex is observed, which is isostructural with [M(nPr-BTP)3]3+ (M = Am, Ln) complexes studied earlier. In contrast, 1H, 13C, and 15N NMR spectra recorded in polar protic solvents showed twice as many signals, indicating a breakdown of symmetry. Supported by density functional theory (DFT) calculations, this difference is explained by the solvent effect on the steric arrangement of the propyl moieties located on the triazine rings. Important information on bonding properties was obtained by 15N NMR. In contrast to the respective Am(III) complex showing a significant covalent contribution, the Th(IV)-BTP interaction is mainly electrostatic.

Characterization of six Merkel cell polyomavirus-positive Merkel cell carcinoma cell lines: Integration pattern suggest that large T antigen truncating events occur before or during integration.

Merkel cell carcinoma (MCC), an aggressive neuroendocrine skin tumor, is a polyomavirus-induced human cancer. To study the causal relationship of MCC carcinogenesis with the integrated Merkel cell polyomavirus (MCPyV) in detail, well-characterized MCC cell lines are needed. Consequently, in the current study, we established and characterized six MCPyV-positive MCC cell lines. Microarray-based comparative genomic hybridization revealed a stable genome carrying only a limited number of chromosomal gains and deletions. All cell lines expressed MCC markers Keratin-20 and neuron-specific enolase as well as truncated MCPyV-encoded large T antigen (LT). For five cell lines, we were able to identify the MCPyV-integration sites in introns of different genes. The LT-truncating stop codon mutations and integration sites were affirmed in the respective clinical patient samples. Inverse PCR suggested that three of the cell lines contained MCPyV genomes as concatemers. This notion was confirmed for the two cell lines with known integration sites. Importantly, our observation of distinct stop codon mutations in cell lines with concatemeric MCPyV integration indicates that these LT-truncating mutations occur before integration. In summary, we provide the detailed characterization of six MCPyV-positive MCC cell lines, which are likely to serve as valuable tools in future MCC research.

Activation of the Aromatic Core of 3,3'-(Pyridine-2,6-diylbis(1H-1,2,3-triazole-4,1-diyl))bis(propan-1-ol)-Effects on Extraction Performance, Stability Constants, and Basicity.

The "CHON" compatible water-soluble ligand 3,3'-(pyridine-2,6-diylbis(1H-1,2,3-triazole-4,1-diyl))bis(propan-1-ol) (PTD) has shown promise for selectively stripping actinide ions from an organic phase containing both actinide and lanthanide ions, by preferential complexation of the former. Aiming at improving its complexation properties, PTD-OMe was synthesized, bearing a methoxy group on the central pyridine ring, thus increasing its basicity and hence complexation strength. Unfortunately, solvent extraction experiments in the range of 0.1-1 mol/L nitric acid proved PTD-OMe to be less efficient than PTD. This behavior is explained by its greater pKa value (pKa = 2.54) compared to PTD (pKa = 2.1). This counteracts its improved complexation properties for Cm(III) (log ß3(PTD-OMe) = 10.8 ± 0.4 versus log ß3(PTD) = 9.9 ± 0.5).

Improved metal allergen reactivity of artificial skin models by integration of Toll-like receptor 4-positive cells.

BACKGROUND: Reconstructed human epidermis (RhE) is widely used to replace animal models in order to assess the proinflammatory and allergenic effects of chemicals. Unfortunately, RhE lacks proinflammatory responsiveness for metal haptens, which are the most prevalent human contact allergens, raising concerns about its reliability for predicting skin allergens. OBJECTIVES: To investigate whether this limitation of RhE might be attributable to a lack of functional expression of Toll-like receptor 4 (TLR4), which governs proinflammatory sensitivity to nickel and cobalt. MATERIALS AND METHODS: RhE, dendritic cell (DC)-containing RhE and full-thickness skin equivalent (FTSE) were compared regarding their proinflammatory responsiveness to metal allergens. RESULTS: The incorporation of dermal fibroblasts was sufficient to confer metal sensitivity to RhE. Unlike keratinocytes, normal human fibroblasts expressed high levels of TLR4 mRNA and induced interleukin-8 expression upon stimulation with nickel or cobalt. Consistently, dermal isolates from FTSE expressed considerable amounts of TLR4 mRNA, whereas RhE or epidermis isolated from FTSE, normal human epidermis or inflamed human epidermis failed to express TLR4. Similarly, co-culture with TLR4-positive DCs bestowed RhE with proinflammatory responsiveness to metals. CONCLUSION: Our data suggest that FTSE or DC/RhE co-culture models can circumvent the shortcomings of RhE assays, and combine the benefits of complex and monoculture-based test systems in a single assay.

Potent NLRP3 Inflammasome Activation by the HIV Reverse Transcriptase Inhibitor Abacavir.

There is experimental and clinical evidence that some exanthematous allergic drug hypersensitivity reactions are mediated by drug-specific T cells. We hypothesized that the capacity of certain drugs to directly stimulate the innate immune system may contribute to generate drug-specific T cells. Here we analyzed whether abacavir, an HIV-1 reverse transcriptase inhibitor often inducing severe delayed-type drug hypersensitivity, can trigger innate immune activation that may contribute to its allergic potential. We show that abacavir fails to generate direct innate immune activation in human monocytes but potently triggers IL-1ß release upon pro-inflammatory priming with phorbol ester or Toll-like receptor stimulation. IL-1ß processing and secretion were sensitive to Caspase-1 inhibition, NLRP3 knockdown, and K+ efflux inhibition and were not observed with other non-allergenic nucleoside reverse transcriptase inhibitors, identifying abacavir as a specific inflammasome activator. It further correlated with dose-dependent mitochondrial reactive oxygen species production and cytotoxicity, indicating that inflammasome activation resulted from mitochondrial damage. However, both NLRP3 depletion and inhibition of K+ efflux mitigated abacavir-induced mitochondrial reactive oxygen species production and cytotoxicity, suggesting that these processes were secondary to NLRP3 activation. Instead, depletion of cardiolipin synthase 1 abolished abacavir-induced IL-1ß secretion, suggesting that mitochondrial cardiolipin release may trigger abacavir-induced inflammasome activation. Our data identify abacavir as a novel inflammasome-stimulating drug allergen. They implicate a potential contribution of innate immune activation to medication-induced delayed-type hypersensitivity, which may stimulate new concepts for treatment and prevention of drug allergies.

NMReDATA, a standard to report the NMR assignment and parameters of organic compounds.

Even though NMR has found countless applications in the field of small molecule characterization, there is no standard file format available for the NMR data relevant to structure characterization of small molecules. A new format is therefore introduced to associate the NMR parameters extracted from 1D and 2D spectra of organic compounds to the proposed chemical structure. These NMR parameters, which we shall call NMReDATA (for nuclear magnetic resonance extracted data), include chemical shift values, signal integrals, intensities, multiplicities, scalar coupling constants, lists of 2D correlations, relaxation times, and diffusion rates. The file format is an extension of the existing Structure Data Format, which is compatible with the commonly used MOL format. The association of an NMReDATA file with the raw and spectral data from which it originates constitutes an NMR record. This format is easily readable by humans and computers and provides a simple and efficient way for disseminating results of structural chemistry investigations, allowing automatic verification of published results, and for assisting the constitution of highly needed open-source structural databases.

Phosphorus in recycling fertilizers - analytical challenges.

The importance of secondary raw materials for phosphorus (P) fertilizer production is expected to increase in the future due to resource depletion, supply risks, and heavy metal contamination of fossil phosphate resources. Municipal wastewater is a promising source for P recovery. In Germany for instance, it contains almost 50% of the total amount of P that is currently applied as mineral fertilizer. Several procedures have been developed to recover and re-use P resulting in a growing number of recycling fertilizers that are currently not regulated in terms of fertilizer efficiency. We tested various materials and matrices for their total P content, solubility of P in neutral ammonium citrate (Pnac) and water, and performed robustness tests to check if existing analytical methods are suitable for those new materials. Digestion with inverse aqua regia was best suited to determine the total P content. Pnac sample preparation and analyses were feasible for all matrices. However, we found significant time and temperature dependencies, especially for materials containing organic matter. Furthermore, several materials didn't reach equilibrium during the extractions. Thus, strict compliance of the test conditions is strongly recommended to achieve comparable results.

Serine 220 phosphorylation of the Merkel cell polyomavirus large T antigen crucially supports growth of Merkel cell carcinoma cells.

Merkel cell polyomavirus (MCPyV) is regarded as a major causal factor for Merkel cell carcinoma (MCC). Indeed, tumor cell growth of MCPyV-positive MCC cells is dependent on the expression of a truncated viral Large T antigen (LT) with an intact retinoblastoma protein (RB)-binding site. Here we determined the phosphorylation pattern of a truncated MCPyV-LT characteristically for MCC by mass spectrometry revealing MCPyV-LT as multi-phospho-protein phosphorylated at several serine and threonine residues. Remarkably, disruption of most of these phosphorylation sites did not affect its ability to rescue knockdown of endogenous T antigens in MCC cells indicating that phosphorylation of the respective amino acids is not essential for the growth promoting function of MCPyV-LT. However, alteration of serine 220 to alanine completely abolished the ability of MCPyV-LT to support proliferation of MCC cells. Conversely, mimicking the phosphorylated state by mutation of serine 220 to glutamic acid resulted in a fully functional LT. Moreover, MCPyV-LT(S220A) demonstrated reduced binding to RB in co-immunoprecipitation experiments as well as weaker induction of RB target genes in MCC cells. In conclusion, we provide evidence that phosphorylation of serine 220 is required for efficient RB inactivation in MCC and may therefore be a potential target for future therapeutic approaches.

CD40-independent natural killer-cell help promotes dendritic cell vaccine-induced T-cell immunity against endogenous B-cell lymphoma.

It is well established that an interplay between natural killer (NK) cells and dendritic cells (DCs) gives rise to their reciprocal activation and provides a Th1-biased cytokine milieu that fosters antitumor T-cell responses. Ex vivo-differentiated DCs transferred into mice strongly stimulate endogenous NK cells to produce interferon (IFN)-γ and initiate a cascade that eventually leads to cytotoxic T-lymphocyte responses. We show that the ability of exogenous DCs to trigger this pathway obviates CD40 signaling and CD4(+) T-cell help and depends on a preceding maturation step. Importantly, this mechanism was also effective in endogenously arising tumors where IFN-γ production is compromised in contrast to transplantable tumors. In c-myc-transgenic mice developing spontaneous lymphomas, injection of unpulsed DCs caused NK-cell activation and induced CD8(+) T cells capable of recognizing the lymphoma cells. Animals treated with unpulsed DCs showed a survival benefit compared to untreated myc mice. Hence, tumor immunity induced by DC-based vaccines not only depends on specific antigens loaded on the DCs. Rather, DC vaccines generate broader immune responses, because endogenous DCs presenting tumor antigens may also become stimulated by NK cells that were activated by exogenous DCs. Thus, the DC/NK-cell/cytotoxic T lymphocyte axis may commonly have relevance for DC-based vaccination protocols in clinical settings.

Complete survey of German sewage sludge ash.

The amount of sewage sludge produced worldwide is expected to further increase due to rising efforts in wastewater treatment. There is a growing concern against its direct use as fertilizer due to contamination of the sludge with heavy metals and organic pollutants. Incinerating the sludge degrades organic compounds almost completely and concentrates heavy metals and phosphorus. However, the sewage sludge ash (SSA) is almost completely disposed of and with it all resources are removed from the economic cycle. Comprehensive knowledge of the composition of SSA is crucial to assess the resource recovery potentials. We conducted a survey of all SSA emerging in Germany and determined the respective mass fractions of 57 elements over a period of one year. The median content of phosphorus was 7.9%, indicating an important recovery potential. Important trace elements were Zn (2.5 g/kg), Mn (1.3 g/kg), and Cu (0.9 g/kg). Mass fractions of technology metals such as V, Cr, Ga, Nb, and rare earths were comparatively low. Considering the possible use of SSA as secondary raw material for fertilizer production it should be noted that its Cd and U content (2.7 mg/kg and 4.9 mg/kg respectively) is significantly lower than that of rock phosphate based mineral fertilizers.

Selective removal of zinc and lead from electric arc furnace dust by chlorination-evaporation reactions.

Re-melting of scrap in an electric arc furnace (EAF) results in the accumulation of filter dust from off-gas treatment that predominantly consists of iron and zinc oxides. Filter dust is classified as hazardous waste due to its high contents of potentially toxic or ecotoxic elements such as Pb, Cr, Cd, and As. A promising processing route for this waste is selective chlorination, in which the non-ferrous metal oxides are chlorinated and selectively evaporated in form of their respective chlorides from the remaining solids via the process gas flow. Here, we investigate stepwise thermochemical treatment of EAF dust with either waste iron(II) chloride solution or hydrochloric acid at 650, 800, and 1100 °C. The Zn and Pb contents of the thermochemically processed EAF dust could be lowered from 29.9% and 1.63% to 0.09% and 0.004%, respectively. Stepwise heating allowed high separation between zinc chloride at the 650 °C step and sodium-, potassium-, and lead-containing chlorides at higher temperatures. Furthermore, the lab-scale results were transferred to the use of an experimental rotary kiln highlighting the possibilities of upscaling the presented process. Selective chlorination of EAF dust with liquid chlorine donors is, therefore, suggested as a potential recycling method for Zn-enriched steelworks dusts.

Heavy metal removal from sewage sludge ash by thermochemical treatment with polyvinylchloride.

Sewage sludge ash (SSA) is a prospective phosphorus source for the future production of recycling P-fertilizers. Due to its high heavy metals contents and the relatively low P plant-availability, SSA must be treated before agricultural utilisation. In this paper SSA was thermochemically treated with PVC in a bench-scale rotary furnace in order to remove heavy metals via the chloride pathway. PVC has a high Cl-content of 52-53% and a high heating value that can be beneficially used for the thermochemical process. Large amounts of waste PVC are already recovered in recycling processes, but there are still some fractions that would be available for the proposed thermochemical process, for example, the low quality near-infrared(NIR)-fraction from waste separation facilities. Heavy metals were effectively removed at temperatures in the range of 800-950 °C via the gas phase by utilisation of PVC as Cl-donor. The resulting P plant-availability was comparable to SSA thermochemically treated with MgCl(2) as Cl-donor if MgO was used as an additive (Mg-donor). A further increase of the plant availability of phosphorus was achieved by acid post-treatment of the thermochemically treated SSA.

4-[(5-Methyl-1H-pyrazol-3-yl)amino]-2H-phenyl-1-phthalazinone Inhibits MCPyV T Antigen Expression in Merkel Cell Carcinoma Independent of Aurora Kinase A.

Merkel cell carcinoma (MCC) is frequently caused by the Merkel cell polyomavirus (MCPyV), and MCPyV-positive tumor cells depend on expression of the virus-encoded T antigens (TA). Here, we identify 4-[(5-methyl-1H-pyrazol-3-yl)amino]-2H-phenyl-1-phthalazinone (PHT)-a reported inhibitor of Aurora kinase A-as a compound inhibiting growth of MCC cells by repressing noncoding control region (NCCR)-controlled TA transcription. Surprisingly, we find that TA repression is not caused by inhibition of Aurora kinase A. However, we demonstrate that ß-catenin-a transcription factor repressed by active glycogen synthase kinase 3 (GSK3)-is activated by PHT, suggesting that PHT bears a hitherto unreported inhibitory activity against GSK3, a kinase known to function in promoting TA transcription. Indeed, applying an in vitro kinase assay, we demonstrate that PHT directly targets GSK3. Finally, we demonstrate that PHT exhibits in vivo antitumor activity in an MCC xenograft mouse model, suggesting a potential use in future therapeutic settings for MCC.

An intact retinoblastoma protein-binding site in Merkel cell polyomavirus large T antigen is required for promoting growth of Merkel cell carcinoma cells.

Merkel cell carcinoma (MCC) is a highly aggressive skin cancer that frequently harbours Merkel cell polyomavirus (MCV) DNA integrated in the genome of the tumor cells. In our study, we elaborate our recent finding that MCV-positive MCC cell lines require the expression of the viral T antigens (TA). Indeed, in a xeno-transplantation model, we prove that TA expression is essential also in an in vivo situation, as knock down of TA leads to tumor regression. Moreover, rescuing TA short hairpin RNA (shRNA)-treated MCV-positive MCC cells by ectopic expression of shRNA-insensitive TAs clearly demonstrates that the observed effect is caused by TA knockdown. Notably, introduction of a mutation in the LTA protein interfering with LTA binding to the retinoblastoma protein (RB) ablated this rescue. The importance of this interaction was further confirmed as LTA-specific knockdown leads to explicit cell growth inhibition. In summary, the presented data demonstrate that established MCV-positive MCC tumors critically depend on TA expression, in particular the LTA and RB interaction, for sustained tumor growth. Consequently, interference with LTA/RB interaction appears as promising strategy to treat MCC.