Nanoscale intermittent contact-scanning electrochemical microscopy

A major theme in scanning electrochemical microscopy (SECM) is a methodology for nanoscale imaging with distance control and positional feedback of the tip. We report the expansion of intermittent contact (IC)-SECM to the nanoscale, using disk-type Pt nanoelectrodes prepared using the laser-puller sealing method. The Pt was exposed using a focused ion beam milling procedure to cut the end of the electrode to a well-defined glass sheath radius, which could also be used to reshape the tips to reduce the size of the glass sheath. This produced nanoelectrodes that were slightly recessed, which was optimal for IC-SECM on the nanoscale, as it served to protect the active part of the tip. A combination of finite element method simulations, steady-state voltammetry and scanning electron microscopy for the measurement of critical dimensions, was used to estimate Pt recession depth. With this knowledge, the tip-substrate alignment could be further estimated by tip approach curve measurements. IC-SECM has been implemented by using a piezo-bender actuator for the detection of damping of the oscillation amplitude of the tip, when IC occurs, which was used as a tip-position feedback mechanism. The piezo-bender actuator improves significantly on the performance of our previous setup for IC-SECM, as the force acting on the sample due to the tip is greatly reduced, allowing studies with more delicate tips. The capability of IC-SECM is illustrated with studies of a model electrode (metal/glass) substrate

Characterization of nanopipettes

Nanopipettes are widely used in electrochemical and analytical techniques as tools for sizing, sequencing, sensing, delivery and imaging. For all of these applications, the response of a nanopipette is strongly affected by its geometry and surface chemistry. As the size of nanopipettes becomes smaller, precise geometric characterization is increasingly important, especially if nanopipette probes are to be used for quantitative studies and analysis. This contribution highlights the combination of data from voltage-scanning ion conductivity experiments, transmission electron microscopy (TEM) and finite element method (FEM) simulations to fully characterize nanopipette geometry and surface charge characteristics, with an accuracy not achievable using existing approaches. Indeed, it is shown that presently used methods for nanopipette characterization can lead to highly erroneous information on nanopipettes. The new approach to characterization further facilitates high-level quantification of the behavior of nanopipettes in electrochemical systems, as demonstrated herein for a scanning ion conductance microscope (SICM) setup

Developments and applications of electrochemical microscopy

This thesis is concerned with the use of electrochemical microscopy, in particular the development and application of the scanning electrochemical microscope (SECM). The concept of intermittent contact (IC), i.e. detecting the oscillation amplitude damping of an SECM imaging probe as it makes physical contact with a surface, is presented as a non-electrochemical means to provide tip-substrate distance feedback, in IC-SECM. This is briefly demonstrated for localised etch pit formation on a calcite crystal. A new imaging mode that incorporates a hopping imaging mode with the principles of intermittent contact is demonstrated for a range of samples, in hopping (H)IC-SECM. HIC-SECM uses an oscillating probe, so alternating current data are also obtained, and this type of scanning mode allows three-dimensional visualisation of the flux around an interface, which greatly enhances the information content compared to other types of electrochemical imaging. The resolution achievable by constant-distance imaging using IC-SECM is greater than constant-height imaging, although the dimensions of the electrode used in imaging will limit the resolution. Pt disk nanoelectrodes were also prepared, that were milled using focussed ion beam-scanning electron microscopy (FIB-SEM). These probes were used to image a model substrate, a gold band on glass, using IC-SECM in a low-force soft-tapping setup. The extension of this work to electrochemical systems focuses on studies of electrodeposition of silver nanoparticles on basal plane highly oriented pyrolytic graphite (HOPG). A nucleation-aggregative growth-detachment mechanism is proposed as an important feature of the process, through both macroscopic and microscopic scanning electrochemical cell microscopy (SECCM). The active sites for electron transfer on HOPG, a topic of recent debate, were also probed in macroscopic and microscopic experiments for silver electrodeposition. The difficulties of working on the electrochemistry of quinones in non-aqueous media are highlighted, with particular focus on methods to overcome the issue of electrode fouling. Different degrees of surface fouling were observed depending both on the electrode material, the compound studied and its concentration. IC-SECM was introduced as a means of establishing a close tip-substrate gap without needing to do electrochemistry for positioning and thus minimising fouling effects, from which high rate constants could be measured and the effect of electrode material on electron transfer kinetics investigated

The validity of the Distress Thermometer in female partners of men with prostate cancer

© 2018 John Wiley & Sons Ltd Female partners of prostate cancer (PCa) survivors experience heightened psychological distress that may be greater than that expressed by PCa patients. However, optimal approaches to detect distressed, or at risk of distress, partners are unclear. This study applied receiver operating characteristics analysis to evaluate diagnostic accuracy, sensitivity and specificity of the Distress Thermometer (DT) compared to widely used measures of general (Hospital Anxiety and Depression Scale) and cancer-specific (Impact of Events Scale-Revised) distress. Participants were partners of men with localised PCa (recruited around diagnosis) about to undergo or had received surgical treatment (N = 189), and partners of men diagnosed with PCa who were 2–4 years post-treatment (N = 460). In both studies, diagnostic utility of the DT overall was not optimal. Although area under the curve scores were acceptable (ranges: 0.71–0.92 and 0.83–0.94 for general and cancer-specific distress, respectively), sensitivity, specificity and optimal DT cut-offs for partner distress varied for general (range: ≥2 to ≥5) and cancer-specific (range: ≥3 to ≥5) distress both across time and between cohorts. Thus, it is difficult to draw firm conclusions about the diagnostic capabilities of the DT for partners or recommend its use in this population. More comprehensive screening measures may be needed to detect partners needing psychological intervention

Time-resolved detection and analysis of single nanoparticle electrocatalytic impacts

There is considerable interest in understanding the interaction and activity of single entities, such as (electro)catalytic nanoparticles (NPs), with (electrode) surfaces. Through the use of a high bandwidth, high signal/noise measurement system, NP impacts on an electrode surface are detected and analyzed in unprecedented detail, revealing considerable new mechanistic information on the process. Taking the electrocatalytic oxidation of H2O2 at ruthenium oxide (RuOx) NPs as an example, the rise time of current–time transients for NP impacts is consistent with a hydrodynamic trapping model for the arrival of a NP with a distance-dependent NP diffusion-coefficient. NP release from the electrode appears to be aided by propulsion from the electrocatalytic reaction at the NP. High-frequency NP impacts, orders of magnitude larger than can be accounted for by a single pass diffusive flux of NPs, are observed that indicate the repetitive trapping and release of an individual NP that has not been previously recognized. The experiments and models described could readily be applied to other systems and serve as a powerful platform for detailed analysis of NP impacts

The HSP90 inhibitor ganetespib: a potential effective agent for Acute Myeloid Leukemia in combination with cytarabine

HSP90 is a multi-client chaperone involved in regulating a large array of cellular processes and is commonly overexpressed in many different cancer types including hematological malignancies. Inhibition of HSP90 holds promise for targeting multiple molecular abnormalities and is therefore an attractive target for heterogeneous malignancies such as Acute Myeloid Leukemia (AML). Ganetespib is a highly potent second generation HSP90 inhibitor which we show is significantly more effective against primary AML blasts at nanomolar concentrations when compared with cytarabine (p < 0.001). Dose dependant cytotoxicity was observed with an apoptotic response coordinate with the loss of pro-survival signaling through the client protein AKT. Combination treatment of primary blasts with ganetespib and cytarabine showed good synergistic interaction (combination index (CI): 0.47) across a range of drug effects with associated reduction in HSP70 feedback and AKT signaling levels. In summary, we show ganetespib to have high activity in primary AMLs as a monotherapy and a synergistic relationship with cytarabine when combined. The combination of cytotoxic cell death, suppression of cytoprotective/drug resistance mechanisms such as AKT and reduced clinical toxicity compared to other HSP90 inhibitors provide strong rationale for the clinical assessment of ganetespib in AML

CD4+ T Cells Reactive to Enteric Bacterial Antigens in Spontaneously Colitic C3H/HeJBir Mice: Increased T Helper Cell Type 1 Response and Ability to Transfer Disease

C3H/HeJBir mice are a new substrain that spontaneously develop colitis early in life. This study was done to determine the T cell reactivity of C3H/HeJBir mice to candidate antigens that might be involved in their disease. C3H/HeJBir CD4+ T cells were strongly reactive to antigens of the enteric bacterial flora, but not to epithelial or food antigens. The stimulatory material in the enteric bacteria was trypsin sensitive and restricted by class II major histocompatibility complex molecules, but did not have the properties of a superantigen. The precursor frequency of interleuken (IL)-2–producing, bacterial-reactive CD4+ T cells in colitic mice was 1 out of 2,000 compared to 1 out of 20,000–25,000 in noncolitic control mice. These T cells produced predominately IL-2 and interferon γ, consistent with a T helper type 1 cell response and were present at 3–4 wk, the age of onset of the colitis. Adoptive transfer of bacterial-antigen–activated CD4+ T cells from colitic C3H/HeJBir but not from control C3H/HeJ mice into C3H/HeSnJ scid/scid recipients induced colitis. These data represent a direct demonstration that T cells reactive with conventional antigens of the enteric bacterial flora can mediate chronic inflammatory bowel disease

Hopping intermittent contact-scanning electrochemical microscopy (HIC-SECM) as a new local dissolution kinetic probe : application to salicylic acid dissolution in aqueous solution

Dissolution kinetics of the (110) face of salicylic acid in aqueous solution is determined by hopping intermittent contact-scanning electrochemical microscopy (HIC-SECM) using a 2.5 μm diameter platinum ultramicroelectrode (UME). The method operates by translating the probe UME towards the surface at a series of positions across the crystal and inducing dissolution via the reduction of protons to hydrogen, which titrates the weak acid and promotes the dissolution reaction, but only when the UME is close to the crystal. Most importantly, as dissolution is only briefly and transiently induced at each location, the initial dissolution kinetics of an as-grown single crystal surface can be measured, rather than a surface which has undergone significant dissolution (pitting), as in other techniques. Mass transport and kinetics in the system are modelled using finite element method simulations which allows dissolution rate constants to be evaluated. It is found that the kinetics of an ‘as-grown’ crystal are much slower than for a surface that has undergone partial bulk dissolution (mimicking conventional techniques), which can be attributed to a dramatic change in surface morphology as identified by atomic force microscopy (AFM). The ‘as-grown’ (110) surface presents extended terrace structures to the solution which evidently dissolve slowly, whereas a partially dissolved surface has extensive etch features and step sites which greatly enhance dissolution kinetics. This means that crystals such as salicylic acid will show time-dependent dissolution kinetics (fluxes) that are strongly dependent on crystal history, and this needs to be taken into account to fully understand dissolution

Language motivation in a reconfigured Europe: access, identity, autonomy

In this paper, I propose that we need to develop an appropriate set of conceptual tools for examining motivational issues pertaining to linguistic diversity, mobility and social integration in a rapidly changing and expanding Europe. I begin by drawing on research that has begun to reframe the concept of integrative motivation in the context of theories of self and identity. Expanding the notion of identity, I discuss the contribution of the Council of Europe's European Language Portfolio in promoting a view of motivation as the development of a plurilingual European identity and the enabling of access and mobility across a multilingual Europe. Next, I critically examine the assumption that the individual pursuit of a plurilingual identity is unproblematic, by highlighting the social context in which motivation and identity are constructed and embedded. To illuminate the role of this social context, I explore three inter-related theoretical frameworks: poststructuralist perspectives on language motivation as 'investment'; sociocultural theory; and theories of autonomy in language education. I conclude with the key message that, as with autonomy, language motivation today has an inescapably political dimension of which we need to take greater account in our research and pedagogical practice

Combination of a mitogen‐activated protein kinase inhibitor with the tyrosine kinase inhibitor pacritinib combats cell adhesion‐based residual disease and prevents re‐expansion of FLT3 ‐ITD acute myeloid leukaemia

Minimal residual disease (MRD) in acute myeloid leukaemia (AML) poses a major challenge due to drug insensitivity and high risk of relapse. Intensification of chemotherapy and stem cell transplantation are often pivoted on MRD status. Relapse rates are high even with the integration of first‐generation FMS‐like tyrosine kinase 3 (FLT3) inhibitors in pre‐ and post‐transplant regimes and as maintenance in FLT3 ‐mutated AML. Pre‐clinical progress is hampered by the lack of suitable modelling of residual disease and post‐therapy relapse. In the present study, we investigated the nature of pro‐survival signalling in primary residual tyrosine kinase inhibitor (TKI)‐treated AML cells adherent to stroma and further determined their drug sensitivity in order to inform rational future therapy combinations. Using a primary human leukaemia‐human stroma model to mimic the cell–cell interactions occurring in patients, the ability of several TKIs in clinical use, to abrogate stroma‐driven leukaemic signalling was determined, and a synergistic combination with a mitogen‐activated protein kinase (MEK) inhibitor identified for potential therapeutic application in the MRD setting. The findings reveal a common mechanism of stroma‐mediated resistance that may be independent of mutational status but can be targeted through rational drug design, to eradicate MRD and reduce treatment‐related toxicity