Transient Silencing of a Type IV P-Type ATPase, Atp10c, Results in Decreased Glucose Uptake in C2C12 Myotubes

Atp10c is a strong candidate gene for diet-induced obesity and type 2 diabetes. To identify molecular and cellular targets of ATP10C, Atp10c expression was altered in vitro in C2C12 skeletal muscle myotubes by transient transfection with an Atp10c-specific siRNA. Glucose uptake assays revealed that insulin stimulation caused a significant 2.54-fold decrease in 2-deoxyglucose uptake in transfected cells coupled with a significant upregulation of native mitogen-activated protein kinases (MAPKs), p38, and p44/42. Additionally, glucose transporter-1 (GLUT1) was significantly upregulated; no changes in glucose transporter-4 (GLUT4) expression were observed. The involvement of MAPKs was confirmed using the specific inhibitor SB203580, which downregulated the expression of native and phosphorylated MAPK proteins in transfected cells without any changes in insulin-stimulated glucose uptake. Results indicate that Atp10c regulates glucose metabolism, at least in part via the MAPK pathway, and, thus, plays a significant role in the development of insulin resistance and type 2 diabetes

Proton Microprobe Analysis of Trace-Element Variations in Vitrinites in the Same and Different Coal Beds

The PIXE (proton-induced X-ray emission) microprobe can be used for nondestructive, in-situ analyses of areas as small as those analyzed by the electron microprobe, and has a sensitivity of detection as much as two orders of magnitude better than the electron microprobe. Preliminary studies demonstrated that PIXE provides a capability for quantitative determination of elemental concentrations in individual coal maceral grains with a detection limit of 1-10 ppm for most elements analyzed. Encouraged by the earlier results, we carried out the analyses reported below to examine trace element variations laterally (over a km range) as well as vertically (cm to m) in the I and J coal beds in the Upper Cretaceous Ferron Sandstone Member of the Mancos Shale in central Utah, and to compare the data with the data from two samples of eastern coals of Pennsylvanian age. The data obtained illustrate the sensitivity of detection obtainable with the PIXE microprobe in analysis of coal macerals. Such data are of value in tracing the geochemical conditions during deposition and diagenesis of a coal bed, and in assessing potential applications and problems of combustion, gasification, or liquefaction of particular coals

Elbow

The Elbow, the part of the human body that a blind or partially sighted person takes hold of, four fingers on the inside, thumb on the outside; holding with a firm grip to assist them with navigation. It is a common guiding technique linking two people into an unfamiliar choreography. In the summer of 2015 C&C Gallery hosted an innovative residency programme, transforming the gallery space into a dynamic studio environment in order to respond to ideas surrounding the condition of blindness. During this time eight artists variously came and went, engaging with a whole repertoire of processes, from casting to laser scanning, leaving a variety of traces of their activities. Much of the work, arrived at through a collaborative approach has sought to obliquely cast a shadow across the subject, remaining open towards the process of making, and the work remains in a state of uncertainty even at the time of writing. During a workshop in the space hosted by John Dickinson-Lilley, the artists wore blind glasses and stumbled rather pathetically around the gallery and the local high street, feeling unsure, insecure and inept, grounded only by the feeling of the floor beneath ones feet and ones fingers upon a companions elbow. Within the space; chalk lines, floor to ceiling columns and laser scanning targets have created an installation for measurement, where diverse material usage- including borax, expanding foam and cat litter humorously confound expectation

Experimental Designs for Binary Data in Switching Measurements on Superconducting Josephson Junctions

We study the optimal design of switching measurements of small Josephson junction circuits which operate in the macroscopic quantum tunnelling regime. Starting from the D-optimality criterion we derive the optimal design for the estimation of the unknown parameters of the underlying Gumbel type distribution. As a practical method for the measurements, we propose a sequential design that combines heuristic search for initial estimates and maximum likelihood estimation. The presented design has immediate applications in the area of superconducting electronics implying faster data acquisition. The presented experimental results confirm the usefulness of the method. KEY WORDS: optimal design, D-optimality, logistic regression, complementary log-log link, quantum physics, escape measurement

Use of a steerable microcatheter during superselective angiography: impact on radiation exposure and procedural efficiency

Background/purpose To study steerable microcatheter (SM) use in moderate and highly difficult vessel selection compared to conventional pre-shaped microcatheter (CM) use. Material and methods An IRB approved, single institution analysis of 40 complex angiographic procedures with and without superselective microcatheter use during an eight-month period in 2017 was performed. Target vessels were deemed moderate or highly difficult to select based on vessel size, tortuosity, and/or angulation during non-selective initial angiography. Data collected included type of microcatheter used (SM or CM), number of microcatheters and microwires used, procedure time, radiation exposure index (dose area product/DAP), target vessel location, and time to target vessel selection (TTVS; time from device placement to vessel selection). Comparison between the SM and CM groups was performed using Wilcoxon test. Results A SM (SwiftNinja, Merit Medical, South Jordan, UT, USA) was used to select 46 vessels in 20 patients. One or more CMs were used in 20 patients to select 34 vessels. Median TTVS, number of microwires used, total procedure time, and DAP (microGray.m2) were 12 vs. 462.5 s (p < 0.0001), 0 vs. 2 (p < 0.001), and 26,948 vs. 30,904 (p = 0.15) in the SM vs. CM groups, respectively. When adjusted for body mass index (BMI) using a linear model for radiation exposure, patients in the SM group had lower radiation exposure than those in the CM group (p = 0.05). Conclusions Utilization of a steerable microcatheter, without or with a guidewire, leads to easier and faster target vessel selection with shorter procedure times in complex vessel anatomy

Union renewal in historical perspective

This article revisits contemporary union renewal/revival debates through comparison with the late 1930s resurgence of trade unionism in the UK’s engineering industry. It is argued that the 1930s union renewal arose from more favourable contextual conditions than those currently obtaining. It was led by political activists, with better-articulated organisation and greater resonance in the working class than their contemporary counterparts, and who were assisted by state policy and pro-worker forces. Conclusions are drawn in relation to current debates

Synchrotron x-ray fluorescence and extended x-ray absorption fine structure analysis

The advent of dedicated synchrotron radiation sources has led to a significant increase in activity in many areas of science dealing with the interaction of x-rays with matter. Synchrotron radiation provides intense, linearly polarized, naturally collimated, continuously tunable photon beams, which are used to determine not only the elemental composition of a complex, polyatomic, dilute material but also the chemical form of the elements with improved accuracy. Examples of the application of synchrotron radiation include experiments in synchrotron x-ray fluorescence (SXRF) analysis and extended x-ray absorption fine structure (EXAFS) analysis. New synchrotron radiation x-ray microprobes for elemental analysis in the parts per billion range are under construction at several laboratories. 76 references, 24 figures

Unsatisfactory gene transfer into bone-resorbing osteoclasts with liposomal transfection systems

BACKGROUND: Bone-resorbing osteoclasts are multinucleated cells that are formed via fusion of their hematopoietic stem cells. Many of the details of osteoclast formation, activation and motility remain unsolved. Therefore, there is an interest among bone biologists to transfect the terminally differentiated osteoclasts and follow their responses to the transgenes in vitro. Severe difficulties in transfecting the large, adherent osteoclasts have been encountered, however, making the use of modern cell biology tools in osteoclast research challenging. Transfection of mature osteoclasts by non-viral gene transfer systems has not been reported. RESULTS: We have systematically screened the usefulness of several commercial DNA transfection systems in human osteoclasts and their mononuclear precursor cell cultures, and compared transfection efficacy to adenoviral DNA transfection. None of the liposome-based or endosome disruption-inducing systems could induce EGFP-actin expression in terminally differentiated osteoclasts. Instead, a massive cell death by apoptosis was found with all concentrations and liposome/DNA-ratios tested. Best transfection efficiencies were obtained by adenoviral gene delivery. Marginal DNA transfection was obtained by just adding the DNA to the cell culture medium. When bone marrow-derived CD34-positive precursor cells were transfected, some GFP-expression was found at the latest 24 h after transfection. Large numbers of apoptotic cells were found and those cells that remained alive, failed to form osteoclasts when cultured in the presence of RANKL and M-CSF, key regulators of osteoclast formation. In comparison, adenoviral gene delivery resulted in the transfection of CD34-positive cells that remained GFP-positive for up to 5 days and allowed osteoclast formation. CONCLUSION: Osteoclasts and their precursors are sensitive to liposomal transfection systems, which induce osteoclast apoptosis. Gene transfer to mononuclear osteoclast precursors or differentiated osteoclasts was not possible with any of the commercial transfection systems tested. Osteoclasts are non-dividing, adherent cells that are difficult to grow as confluent cultures, which may explain problems with transfection reagents. Large numbers of α(v)β(3 )integrin on the osteoclast surface allows adenovirus endocytosis and infection proceeds in dividing and non-dividing cells efficiently. Viral gene delivery is therefore currently the method of choice for osteoclast transfection

Connexin-mimetic peptide Gap 27 decreases osteoclastic activity

BACKGROUND: Bone remodelling is dependent on the balance between bone resorbing osteoclasts and bone forming osteoblasts. We have shown previously that osteoclasts contain gap-junctional protein connexin-43 and that a commonly used gap-junctional inhibitor, heptanol, can inhibit osteoclastic bone resorption. Since heptanol may also have some unspecific effect unrelated to gap-junctional inhibition we wanted to test the importance of gap-junctional communication to osteoclasts using a more specific inhibitor. METHODS: A synthetic connexin-mimetic peptide, Gap 27, was used to evaluate the contribution of gap-junctional communication to osteoclastic bone resorption. We utilised the well-characterised pit-formation assay to study the effects of the specific gap-junctional inhibitor to the survival and activity of osteoclasts. RESULTS: Gap 27 caused a remarked decrease in the number of both TRAP-positive mononuclear and multinucleated rat osteoclasts cultured on bovine bone slices. The decrease in the cell survival seemed to be restricted to TRAP-positive cells, whereas the other cells of the culture model seemed unaffected. The activity of the remaining osteoclasts was found to be diminished by measuring the percentage of osteoclasts with actin rings of all TRAP-positive cells. In addition, the resorbed area in the treated cultures was greatly diminished. CONCLUSIONS: On the basis of these results we conclude that gap-junctional communication is essential for the action of bone resorbing osteoclasts and for proper remodelling for bone

Different expressions of the same mode: a recent dialogue between archaeological and contemporary drawing practices

In this article we explore what we perceive as pertinent features of shared experience at the excavations of an Iron Age Hillfort at Bodfari, North Wales, referencing artist, archaeologist and examples of seminal art works and archaeological records resulting through inter-disciplinary collaboration. We explore ways along which archaeological and artistic practices of improvisation become entangled and productive through their different modes of mark-making. We contend that marks and memories of artist and archaeologist alike emerge interactively, through the mutually constituting effects of the object of study, the tools of exploration, and the practitioners themselves, when they are enmeshed in the cross-modally bound activities. These include, but are not limited to, remote sensing, surveying, mattocking, trowelling, drawing, photographing, videoing and sound recording. These marks represent the co-signatories: the gesture of the often anonymous practitioners, the voice of the deposits, as well as the imprint of the tools, and their interplay creates a multi-threaded narrative documenting their modes of intra-action, in short our practices. They occupy the conceptual space of paradata, and in the process of saturating the interstices of digital cognitive prosthetics they lend probity to their translations in both art form and archive