Loss of hSef promotes metastasis through upregulation of EMT in prostate cancer

We have previously reported that the negative signaling regulator Similar Expression to FGF (hSef) is downregulated in prostate cancer and its loss is associated with clinical metastasis. Here, we explored the mechanistic basis of this finding. We first confirmed our clinical observation by testing hSef manipulation in an $\textit{in vivo}$ metastasis model. hSef stable expressing cells (PC3M-hSef) or empty vector controls (PC3M-EV) were injected subcutaneously into the lateral thoracic walls of NOD-SCID gamma mice and lungs were harvested at autopsy. In this model, 6/7 PC3M-EV xenografts had definitive lung micro-metastasis whilst only 1/6 PC3M-hSef xenografts exhibited metastasis recapitulating the clinical scenario ($\textit{p}$ = 0.03). Gene expression studies revealed key perturbations in genes involved in cell motility and epithelial to mesenchymal transition (EMT) along with alterations in cognate signaling pathways. These results were validated in an EMT specific PCR array whereby hSef over-expression and silencing reciprocally altered E-Cadherin expression ($\textit{p}$ = <0.001) amongst other EMT markers. Immunohistochemistry of excised tumors from the xenografts also confirmed the effect of hSef in suppressing E-Cadherin expression at the protein level. Phosphokinase arrays further demonstrated a role for hSef in attenuating signaling of not only ERK-MAPK but also the JNK and p38 pathways as well. Taken together, these data suggest evidence that loss of hSef may be a critical event facilitating tumor dissemination of prostate cancer through alteration of EMT. Detection of downregulated hSef, along with other negative regulators, may therefore be a useful biomarker heralding a transition to a metastatic phenotype and warrants further exploration in this context.Grant sponsor: Medical Research Council/The Prostate Cancer Charity Clinical Research Training Fellowship (S.H.); Grant number: G1001961; Grant sponsor: The Royal College of Surgeons Surgical Research Fellowship (S.H.), Addenbrooke’s Charitable Trust Oncology Research Fellowship (S.H.) and the University of Cambridge Raymond and Beverly Sackler Studentship (S.H.) DOI: 10.1002/ijc.30604

Assessing Human Embryonic Stem Cell-Derived Dopaminergic Neuron Progenitor Transplants Using Non-invasive Imaging Techniques

Abstract: Purpose: Human pluripotent stem cell (hPSC)-derived dopaminergic neuron progenitor cells (DAPCs) are a potential therapy for Parkinson’s disease (PD). However, their intracranial administration raises safety concerns including uncontrolled proliferation, migration and inflammation. Here, we apply a bimodal imaging approach to investigate the fate of DAPC transplants in the rat striatum. Procedures: DAPCs co-expressing luciferase and ZsGreen or labelled with micron-sized particles of iron oxide (MPIOs) were transplanted in the striatum of RNU rats (n = 6 per group). DAPCs were tracked in vivo using bioluminescence and magnetic resonance (MR) imaging modalities. Results: Transgene silencing in differentiating DAPCs accompanied with signal attenuation due to animal growth rendered the bioluminescence undetectable by week 2 post intrastriatal transplantation. However, MR imaging of MPIO-labelled DAPCs showed that transplanted cells remained at the site of injection for over 120 days. Post-mortem histological analysis of DAPC transplants demonstrated that labelling with either luciferase/ZsGreen or MPIOs did not affect the ability of cells to differentiate into mature dopaminergic neurons. Importantly, labelled cells did not elicit increased glial reactivity compared to non-labelled cells. Conclusions: In summary, our findings support the transplantation of hPSC-derived DAPCs as a safe treatment for PD

Stem cell culture conditions and stability: a joint workshop of the PluriMes Consortium and Pluripotent Stem Cell Platform

Human stem cells have the potential to transform medicine. However, hurdles remain to ensure that manufacturing processes produce safe and effective products. A thorough understanding of the biological processes occurring during manufacture of what can be very complex products, is fundamental to assuring these qualities and thus, their acceptability to regulators and clinicians. Leaders in both human pluripotent and somatic stem cells, were brought together with experts in clinical translation, bio-manufacturing and regulation, to discuss key issues in assuring appropriate manufacturing conditions for delivery of effective and safe products from these cell types. This report summarizes the key issues discussed and records consensus reached by delegates and emphasizes the need for accurate language and nomenclature in the scientific discourse around stem cells

A reference human induced pluripotent stem cell line for large-scale collaborative studies

Human induced pluripotent stem cell (iPSC) lines are a powerful tool for studying development and disease, but the considerable phenotypic variation between lines makes it challenging to replicate key findings and integrate data across research groups. To address this issue, we sub-cloned candidate human iPSC lines and deeply characterized their genetic properties using whole genome sequencing, their genomic stability upon CRISPR-Cas9-based gene editing, and their phenotypic properties including differentiation to commonly used cell types. These studies identified KOLF2.1J as an all-around well-performing iPSC line. We then shared KOLF2.1J with groups around the world who tested its performance in head-to-head comparisons with their own preferred iPSC lines across a diverse range of differentiation protocols and functional assays. On the strength of these findings, we have made KOLF2.1J and its gene-edited derivative clones readily accessible to promote the standardization required for large-scale collaborative science in the stem cell field

A Critical Review of Mineral Matter Related Issues during Gasification of Coal in Fixed, Fluidized, and Entrained Flow Gasifiers

Gasification of coal is gaining more popularity due to its clean operation, and its ability to generate products for various markets. However, these technologies are not widely commercialized due to reliability and economic issues. Mineral matter in coal plays an important role in affecting the availability/reliability of a gasifier. Agglomeration in the bed, slag mobility and blockage of the syngas exit section are some of the operations related concerns in fixed-bed gasifiers, while ash deposition and sudden defluidization are the major concerns in fluidized bed gasifiers. In the case of entrained flow gasifiers, syngas cooler fouling and blockage, corrosion and erosion of refractory, and slag mobility are some of the major issues affecting the operations and the reliability of the gasifier. This review is aimed at critically examining various mineral matter related issues contributing to the operation and reliability problems in three types of generic gasifiers (fixed bed, fluidized bed and entrained flow gasifiers). Based on the review, some strategies to counter the potential mineral matter related issues are presented

Effect of Temperature, Pressure, Feed Particle Size, and Feed Particle Density on Structural Characteristics and Reactivity of Chars Generated during Gasification of Pittsburgh No.8 Coal in a High-Pressure, High-Temperature Flow Reactor

The gasification behavior of coal under high-temperature and high-pressure conditions is important from the perspective of designing and optimizing high efficiency gasifiers and troubleshooting existing gasifiers. The effect of feed particle size, density, temperature, and pressure on char porous structure, morphology, reflectance, and reactivity under conditions relevant to entrained-flow gasification was investigated. The chars were generated over a range of temperatures (1100, 1300, and 1400 &deg;C at 11.3 bar for the &minus;150 + 106 &micro;m fraction), pressures (3.4, 6.2, 11.3, 15.5, and 21.7 bar at 1300 &deg;C for the &minus;150 + 106 &micro;m fraction), for various size fractions (&minus;106 + 75, &minus;150 + 106, &minus;212 + 150, &minus;420 + 212 &micro;m at 1300 &deg;C and 11.3 bar), and density fractions (&lt;1.3, 1.3&ndash;1.6, &gt;1.6g/cc for the &minus;106 + 75 &micro;m at 1300 &deg;C and 11.3 bar) of Pittsburgh No.8 bituminous coal using a high-pressure, high-temperature flow reactor (HPHTFR) in a equimolar mixture of CO2 and N2. Chars were characterized for conversion, morphology, thermal swelling ratio, and reactivity using ash tracer technique, oil immersion microscopy, tap density technique, and a thermogravimetric analyzer, respectively, and the results were statistically analyzed to determine for effects by feed particle density, feed particle size, temperature, and pressure. The results showed that the conversion was most affected by temperature, followed by feed particle size, pressure, and feed particle density. In the case of structural characteristics (i.e., thermal swelling ratio and group-I char concentration), feed particle density affected group-I concentration, while both feed particle size and feed particle density affected thermal swelling ratio. Variation in vitrinite content and fragmentation affected the thermal swelling ratio and group-I char concentration. In the case of intrinsic reactivity, particle density showed the largest effect, followed by temperature, particle size, and pressure. An increase in reflectance and temperature was found to inversely affect intrinsic reactivity

Effect of Heterogeneity in Coal Ash Chemical Composition on the Onset of Conditions Favorable for Agglomeration in Fluid Beds

Ash agglomeration issues that arise due to the sticking of slag-wetted, colliding particles have been creating operational difficulties and monetary losses for the fluidized bed combustion (FBC) industry. Difficulties have been experienced in the detection of slag-liquid at the low operating temperatures in fluidized bed combustors (FBCs) and predicting the agglomeration behavior of fuel. This study aims to study the effect of heterogeneity in ash composition on the detection of slag-liquid in FBCs. It quantifies the slag-liquid amounts at the particle-level, under oxidizing environments, by dividing the bulk fuel into density classes. FactSage™ thermodynamic simulations of each of the particle classes, along with experimental validation of the trends with thermo-mechanical analysis (TMA) and high temperature X-ray diffraction (HT-XRD) were performed. The results obtained can be used to estimate the stickiness of particles in the development of ash agglomeration models based on particle collisions. The study of these particle classes shows that particle classes with specific minerals can form low temperature eutectics and lead to onset of slag-liquid formation at temperatures below those predicted by bulk analysis alone. Comparison of the differences in slag-liquid formation tendencies under reducing and oxidizing environments is also presented

Influence of Pyrolysis Gas on Volatile Yield and CO2 Reaction Kinetics of the Char Samples Generated in a High-Pressure, High-Temperature Flow Reactor

The influence of pyrolysis atmosphere on volatile yield, structural characteristics, and CO2 reaction kinetics have been examined on chars generated from Pittsburgh No. 8 coal at 6.2 bar pressure and 1100 &deg;C in a high-pressure, high-temperature flow reactor (HPHTFR) in Ar, N2, 50 (vol. %) CO2 and N2 (i.e., CO2/ N2) atmospheres. The chars were characterized for volatile yield, thermal swelling ratio, surface area, pore size distribution, crystallite structure, defects to graphitic intensity ratio, and char-CO2 reactivity. Coal pyrolyzed in CO2/N2 showed higher volatile yield (27%) compared to coal pyrolyzed in argon (~16%) and nitrogen (~19%). Except for volatile yield, there was no significant difference in structural properties for chars generated in different pyrolysis atmospheres. The difference in volatile yield was found to be due to presence of unconverted tetrahydrofuran (THF) soluble tar/soot. The results also showed that the intrinsic reactivity was highest for char generated in N2 atmosphere and lowest for char generated in CO2/N2 atmosphere. The kinetic parameters (activation energy and pre-exponential factor) for the char-CO2 reaction were ascertained using nth order model. The activation energies did not differ significantly among the chars generated in different pyrolysis atmospheres. The order of reaction was found to follow: CO2/N2 char &gt; N2 char &asymp; Ar char