UBE2C expression is elevated in hepatoblastoma and correlates with inferior patient survival.

Hepatoblastoma (HB) is the most common malignant liver tumor among children. To gain insight into the pathobiology of HB, we performed RNA sequence analysis on 5 patient-derived xenograft lines (HB-243, HB-279, HB-282, HB-284, HB-295) and 1 immortalized cell line (HUH6). Using cultured hepatocytes as a control, we found 2,868 genes that were differentially expressed in all of the HB lines on mRNA level. The most upregulated genes were ODAM, TRIM71, and IGDCC3, and the most downregulated were SAA1, SAA2, and NNMT. Protein-protein interaction analysis identified ubiquitination as a key pathway dysregulated in HB. UBE2C, encoding an E2 ubiquitin ligase often overexpressed in cancer cells, was markedly upregulated in 5 of the 6 HB cell lines. Validation studies confirmed UBE2C immunostaining in 20 of 25 HB tumor specimens versus 1 of 6 normal liver samples. The silencing of UBE2C in two HB cell models resulted in decreased cell viability. RNA sequencing analysis showed alterations in cell cycle regulation after UBE2C knockdown. UBE2C expression in HB correlated with inferior patient survival. We conclude that UBE2C may hold prognostic utility in HB and that the ubiquitin pathway is a potential therapeutic target in this tumor.

Integrating Carbon Nanomaterials with Metals for Bio-sensing Applications.

Age structure in most developed countries is changing fast as the average lifespan is increasing significantly, calling for solutions to provide improved treatments for age-related neurological diseases and disorders. In order to address these problems, a reliable way of recording information about neurotransmitters from in vitro and in vivo applications is needed to better understand neurological diseases and disorders as well as currently used treatments. Likewise, recent developments in medicine, especially with the opioid crisis, are demanding a swift move to personalized medicine to administer patient needs rather than population-wide averages. In order to enable the so-called personalized medicine, it is necessary to be able to do measurements in vivo and in real time. These actions require sensitive and selective detection of different analytes from very demanding environments. Current state-of-the-art materials are unable to provide sensitive and selective detection of neurotransmitters as well as the required time resolution needed for drug molecules at a reasonable cost. To meet these challenges, we have utilized different metals to grow carbon nanomaterials and applied them for sensing applications showing that there are clear differences in their electrochemical properties based on the selected catalyst metal. Additionally, we have combined atomistic simulations to support optimizing materials for experiments and to gain further understanding of the atomistic level reactions between different analytes and the sensor surface. With carbon nanostructures grown from Ni and Al + Co + Fe hybrid, we can detect dopamine, ascorbic acid, and uric acid simultaneously. On the other hand, nanostructures grown from platinum provide a feasible platform for detection of H2O2 making them suitable candidates for enzymatic biosensors for detection of glutamate, for example. Tetrahedral amorphous carbon electrodes have an ability to detect morphine, paracetamol, tramadol, and O-desmethyltramadol. With carbon nanomaterial-based sensors, it is possible to reach metal-like properties in sensing applications using only a fraction of the metal as seed for the material growth. We have also seen that by using nanodiamonds as growth catalyst for carbon nanofibers, it is not possible to detect dopamine and ascorbic acid simultaneously, although the morphology of the resulting nanofibers is similar to the ones grown using Ni. This further indicates the importance of the metal selection for specific applications. However, Ni as a continuous layer or as separate islands does not provide adequate performance. Thus, it appears that metal nanoparticles combined with fiber-like morphology are needed for optimized sensor performance for neurotransmitter detection. This opens up a new research approach of application-specific nanomaterials, where carefully selected metals are integrated with carbon nanomaterials to match the needs of the sensing application in question.

Pt-grown carbon nanofibers for detection of hydrogen peroxide.

Removal of left-over catalyst particles from carbon nanomaterials is a significant scientific and technological problem. Here, we present the physical and electrochemical study of application-specific carbon nanofibers grown from Pt-catalyst layers. The use of Pt catalyst removes the requirement for any cleaning procedure as the remaining catalyst particles have a specific role in the end-application. Despite the relatively small amount of Pt in the samples (7.0 ± 0.2%), they show electrochemical features closely resembling those of polycrystalline Pt. In O2-containing environment, the material shows two separate linear ranges for hydrogen peroxide reduction: 1-100 µM and 100-1000 µM with sensitivities of 0.432 µA µM-1 cm-2 and 0.257 µA µM-1 cm-2, respectively, with a 0.21 µM limit of detection. In deaerated solution, there is only one linear range with sensitivity 0.244 µA µM-1 cm-2 and 0.22 µM limit of detection. We suggest that the high sensitivity between 1 µM and 100 µM in solutions where O2 is present is due to oxygen reduction reaction occurring on the CNFs producing a small additional cathodic contribution to the measured current. This has important implications when Pt-containing sensors are utilized to detect hydrogen peroxide reduction in biological, O2-containing environment.

Pt-grown carbon nanofibers for enzymatic glutamate biosensors and assessment of their biocompatibility.

Application-specific carbon nanofibers grown from Pt-catalyst layers have been shown to be a promising material for biosensor development. Here we demonstrate immobilization of glutamate oxidase on them and their use for amperometric detection of glutamate at two different potentials. At -0.15 V vs. Ag/AgCl at concentrations higher than 100 µM the oxygen reduction reaction severely interferes with the enzymatic production of H2O2 and consequently affects the detection of glutamate. On the other hand, at 0.6 V vs. Ag/AgCl enzyme saturation starts to affect the measurement above a glutamate concentration of 100 µM. Moreover, we suggest here that glutamate itself might foul Pt surfaces to some degree, which should be taken into account when designing Pt-based sensors operating at high anodic potentials. Finally, the Pt-grown and Ni-grown carbon nanofibers were shown to be biocompatible. However, the cells on Pt-grown carbon nanofibers had different morphology and formation of filopodia compared to those on Ni-grown carbon nanofibers. The effect was expected to be caused rather by the different fiber dimensions between the samples than the catalyst metal itself. Further experiments are required to find the optimal dimensions of CNFs for biological purposes.

Application-Specific Catalyst Layers: Pt-Containing Carbon Nanofibers for Hydrogen Peroxide Detection.

Complete removal of metal catalyst particles from carbon nanofibers (CNFs) and other carbon nanostructures is extremely difficult, and the envisioned applications may be compromised by the left-over impurities. To circumvent these problems, one should use, wherever possible, such catalyst materials that are meant to remain in the structure and have some application-specific role, making any removal steps unnecessary. Thus, as a proof-of-concept, we present here a nanocarbon-based material platform for electrochemical hydrogen peroxide measurement utilizing a Pt catalyst layer to grow CNFs with intact Pt particles at the tips of the CNFs. Backed by careful scanning transmission electron microscopy analysis, we show that this material can be readily realized with the Pt catalyst layer thickness impacting the resulting structure and also present a growth model to explain the evolution of the different types of structures. In addition, we show by electrochemical analysis that the material exhibits characteristic features of Pt in cyclic voltammetry and it can detect very small amounts of hydrogen peroxide with very fast response times. Thus, the present sensor platform provides an interesting electrode material with potential for biomolecule detection and in fuel cells and batteries. In the wider range, we propose a new approach where the selection of catalytic particles used for carbon nanostructure growth is made so that (i) they do not need to be removed and (ii) they will have essential role in the final application.