Utilization of Microscale Silicon Cantilevers to Assess Cellular Contractile Function In Vitro

The development of more predictive and biologically relevant in vitro assays is predicated on the advancement of versatile cell culture systems which facilitate the functional assessment of the seeded cells. To that end, microscale cantilever technology offers a platform with which to measure the contractile functionality of a range of cell types, including skeletal, cardiac, and smooth muscle cells, through assessment of contraction induced substrate bending. Application of multiplexed cantilever arrays provides the means to develop moderate to high-throughput protocols for assessing drug efficacy and toxicity, disease phenotype and progression, as well as neuromuscular and other cell-cell interactions. This manuscript provides the details for fabricating reliable cantilever arrays for this purpose, and the methods required to successfully culture cells on these surfaces. Further description is provided on the steps necessary to perform functional analysis of contractile cell types maintained on such arrays using a novel laser and photo-detector system. The representative data provided highlights the precision and reproducible nature of the analysis of contractile function possible using this system, as well as the wide range of studies to which such technology can be applied. Successful widespread adoption of this system could provide investigators with the means to perform rapid, low cost functional studies in vitro, leading to more accurate predictions of tissue performance, disease development and response to novel therapeutic treatment

Application of Reservoir Characterization and Advanced Technology to Improve Recovery and Economics in a Lower Quality Shallow Shelf San Andres Reservoir Progress Report

The Class 2 Project at West Welch was designed to demonstrate the use of advanced technologies to enhance the economics of improved oil recovery (IOR) projects in lower quality Shallow Shelf Carbonate (SSC) reservoirs, resulting in recovery of additional oil that would otherwise be left in the reservoir at project abandonment. Accurate reservoir description is critical to the effective evaluation and efficient design of IOR projects in the heterogeneous SSC reservoirs. Therefore, the majority of Budget Period 1 was devoted to reservoir characterization. Technologies being demonstrated include: (1) Advanced petrophysics; (2) Three-dimensional (3-D) seismic; (3) Crosswell bore tomography; (4) Advanced reservoir simulation; (5) Carbon dioxide (CO{sub 2}) stimulation treatments; (6) Hydraulic fracturing design and monitoring; and (7) Mobility control agents

Application of Reservoir Characterization and Advanced Technology to Improve Recovery and Economics in a Lower Quality Shallow Shelf San Andres Reservoir Progress Report

The Class 2 Project at West Welch was designed to demonstrate the use of advanced technologies to enhance the economics of improved oil recovery (IOR) projects in lower quality Shallow Shelf Carbonate (SSC) reservoirs, resulting in recovery of additional oil that would otherwise be left in the reservoir at project abandonment. Accurate reservoir description is critical to the effective evaluation and efficient design of IOR projects in the heterogeneous SSC reservoirs. Therefore, the majority of Budget Period 1 was devoted to reservoir characterization. Technologies being demonstrated include: (1) Advanced petrophysics; (2) Three-dimensional (3-D) seismic; (3) Crosswell bore tomography; (4) Advanced reservoir simulation; (5) Carbon dioxide (CO{sub 2}) stimulation treatments; (6) Hydraulic fracturing design and monitoring; and (7) Mobility control agents

The Family Navigator: A pilot intervention to support intensive care unit family surrogates

Background Although communication problems between family surrogates and intensive care unit (ICU) clinicians have been documented, there are few effective interventions. Nurses have the potential to play an expanded role in ICU communication and decision making. Objectives To conduct a pilot randomized controlled trial of the Family Navigator (FN), a distinct nursing role to address family members’ unmet communication needs early in an ICU stay. Methods An inter-disciplinary team developed the FN protocol. A randomized controlled pilot intervention trial of the FN was performed in a tertiary referral hospital ICU to test the feasibility and acceptability of the intervention. The intervention addressed informational and emotional communication needs through daily contact using structured clinical updates, emotional and informational support modules, family meeting support and follow-up phone calls. Results Twenty-six surrogate/patient pairs (13 per study arm) were enrolled. Surrogates randomized to the intervention had contact with the FN 90% or more of eligible patient days. All surrogates agreed or strongly agreed that they would recommend the FN to other families. Open-ended comments from both surrogates and clinicians were uniformly positive. For both groups, 100% of baseline data collection interviews and 81% of 6–8 week follow-up interviews were completed. Conclusions A fully integrated nurse empowered to facilitate decision making is a feasible intervention in the ICU setting. It is well-received by ICU families and staff. A larger randomized controlled trial is needed to demonstrate an impact on important outcomes, such as surrogate well-being and decision quality

Myths and Methodologies:Standardisation in human physiology research—should we control the controllables?

The premise of research in human physiology is to explore a multifaceted system whilst identifying one or a few outcomes of interest. Therefore, the control of potentially confounding variables requires careful thought regarding the extent of control and complexity of standardisation. One common factor to control prior to testing is diet, as food and fluid provision may deviate from participants’ habitual diets, yet a self‐report and replication method can be flawed by under‐reporting. Researchers may also need to consider standardisation of physical activity, whether it be through familiarisation trials, wash‐out periods, or guidance on levels of physical activity to be achieved before trials. In terms of pharmacological agents, the ethical implications of standardisation require researchers to carefully consider how medications, caffeine consumption and oral contraceptive prescriptions may affect the study. For research in females, it should be considered whether standardisation between‐ or within‐participants in regards to menstrual cycle phase is most relevant. The timing of measurements relative to various other daily events is relevant to all physiological research and so it can be important to standardise when measurements are made. This review summarises the areas of standardisation which we hope will be considered useful to anyone involved in human physiology research, including when and how one can apply standardisation to various contexts

On a higher dimensional version of the Benjamin--Ono equation

We consider a higher dimensional version of the Benjamin--Ono equation, $\partial_t u -\mathcal{R}_1\Delta u+u\partial_{x_1} u=0$, where $\mathcal{R}_1$ denotes the Riesz transform with respect to the first coordinate. We first establish sharp space--time estimates for the associated linear equation. These estimates enable us to show that the initial value problem for the nonlinear equation is locally well-posed in $L^2$-Sobolev spaces $H^{s}(\mathbb{R}^d)$, with $s>5/3$ if $d=2$ and $s>d/2+1/2$ if $d\ge 3$. We also provide ill-posedness results.Comment: We also show that in dimension 2 our results are shar

Identifying the science and technology dimensions of emerging public policy issues through horizon scanning

Public policy requires public support, which in turn implies a need to enable the public not just to understand policy but also to be engaged in its development. Where complex science and technology issues are involved in policy making, this takes time, so it is important to identify emerging issues of this type and prepare engagement plans. In our horizon scanning exercise, we used a modified Delphi technique [1]. A wide group of people with interests in the science and policy interface (drawn from policy makers, policy adviser, practitioners, the private sector and academics) elicited a long list of emergent policy issues in which science and technology would feature strongly and which would also necessitate public engagement as policies are developed. This was then refined to a short list of top priorities for policy makers. Thirty issues were identified within broad areas of business and technology; energy and environment; government, politics and education; health, healthcare, population and aging; information, communication, infrastructure and transport; and public safety and national security.Public policy requires public support, which in turn implies a need to enable the public not just to understand policy but also to be engaged in its development. Where complex science and technology issues are involved in policy making, this takes time, so it is important to identify emerging issues of this type and prepare engagement plans. In our horizon scanning exercise, we used a modified Delphi technique [1]. A wide group of people with interests in the science and policy interface (drawn from policy makers, policy adviser, practitioners, the private sector and academics) elicited a long list of emergent policy issues in which science and technology would feature strongly and which would also necessitate public engagement as policies are developed. This was then refined to a short list of top priorities for policy makers. Thirty issues were identified within broad areas of business and technology; energy and environment; government, politics and education; health, healthcare, population and aging; information, communication, infrastructure and transport; and public safety and national security

Comparative study of in situ N2 rotational Raman spectroscopy methods for probing energy thermalisation processes during spin-exchange optical pumping

Spin-exchange optical pumping (SEOP) has been widely used to produce enhancements in nuclear spin polarisation for hyperpolarised noble gases. However, some key fundamental physical processes underlying SEOP remain poorly understood, particularly in regards to how pump laser energy absorbed during SEOP is thermalised, distributed and dissipated. This study uses in situ ultra-low frequency Raman spectroscopy to probe rotational temperatures of nitrogen buffer gas during optical pumping under conditions of high resonant laser flux and binary Xe/N2 gas mixtures. We compare two methods of collecting the Raman scattering signal from the SEOP cell: a conventional orthogonal arrangement combining intrinsic spatial filtering with the utilisation of the internal baffles of the Raman spectrometer, eliminating probe laser light and Rayleigh scattering, versus a new in-line modular design that uses ultra-narrowband notch filters to remove such unwanted contributions. We report a ~23-fold improvement in detection sensitivity using the in-line module, which leads to faster data acquisition and more accurate real-time monitoring of energy transport processes during optical pumping. The utility of this approach is demonstrated via measurements of the local internal gas temperature (which can greatly exceed the externally measured temperature) as a function of incident laser power and position within the cell

Studying the Underlying Event in Drell-Yan and High Transverse Momentum Jet Production at the Tevatron

We study the underlying event in proton-antiproton collisions by examining the behavior of charged particles (transverse momentum pT > 0.5 GeV/c, pseudorapidity |\eta| < 1) produced in association with large transverse momentum jets (~2.2 fb-1) or with Drell-Yan lepton-pairs (~2.7 fb-1) in the Z-boson mass region (70 < M(pair) < 110 GeV/c2) as measured by CDF at 1.96 TeV center-of-mass energy. We use the direction of the lepton-pair (in Drell-Yan production) or the leading jet (in high-pT jet production) in each event to define three regions of \eta-\phi space; toward, away, and transverse, where \phi is the azimuthal scattering angle. For Drell-Yan production (excluding the leptons) both the toward and transverse regions are very sensitive to the underlying event. In high-pT jet production the transverse region is very sensitive to the underlying event and is separated into a MAX and MIN transverse region, which helps separate the hard component (initial and final-state radiation) from the beam-beam remnant and multiple parton interaction components of the scattering. The data are corrected to the particle level to remove detector effects and are then compared with several QCD Monte-Carlo models. The goal of this analysis is to provide data that can be used to test and improve the QCD Monte-Carlo models of the underlying event that are used to simulate hadron-hadron collisions.Comment: Submitted to Phys.Rev.

The Interdomain Linker of AAV-2 Rep68 Is an Integral Part of Its Oligomerization Domain: Role of a Conserved SF3 Helicase Residue in Oligomerization

The four Rep proteins of adeno-associated virus (AAV) orchestrate all aspects of its viral life cycle, including transcription regulation, DNA replication, virus assembly, and site-specific integration of the viral genome into the human chromosome 19. All Rep proteins share a central SF3 superfamily helicase domain. In other SF3 members this domain is sufficient to induce oligomerization. However, the helicase domain in AAV Rep proteins (i.e. Rep40/Rep52) as shown by its monomeric characteristic, is not able to mediate stable oligomerization. This observation led us to hypothesize the existence of an as yet undefined structural determinant that regulates Rep oligomerization. In this document, we described a detailed structural comparison between the helicase domains of AAV-2 Rep proteins and those of the other SF3 members. This analysis shows a major structural difference residing in the small oligomerization sub-domain (OD) of Rep helicase domain. In addition, secondary structure prediction of the linker connecting the helicase domain to the origin-binding domain (OBD) indicates the potential to form α-helices. We demonstrate that mutant Rep40 constructs containing different lengths of the linker are able to form dimers, and in the presence of ATP/ADP, larger oligomers. We further identified an aromatic linker residue (Y224) that is critical for oligomerization, establishing it as a conserved signature motif in SF3 helicases. Mutation of this residue critically affects oligomerization as well as completely abolishes the ability to produce infectious virus. Taken together, our data support a model where the linker residues preceding the helicase domain fold into an α-helix that becomes an integral part of the helicase domain and is critical for the oligomerization and function of Rep68/78 proteins through cooperative interaction with the OBD and helicase domains