949 research outputs found
Boundary-induced anisotropy of the avalanches in the sandpile automaton
We study numerically the avalanches in a two--dimensional critical height
sandpile model with sand grains added at the center of the system. Smaller
avalanches near the center of the system are isotropic. Larger avalanches are,
however, affected by the boundary of the system, to a degree that increases
with the avalanche size. Up to linear system size , we did not find an
obvious indication for lattice--induced anisotropy.Comment: 7 pages, LaTeX, preprint HLRZ 38/9
A Simple, Highly Efficient Method for Heterologous Expression in Mammalian Primary Neurons Using Cationic Lipid-mediated mRNA Transfection
Expression of heterologous proteins in adult mammalian neurons is a valuable technique for the study of neuronal function. The post-mitotic nature of mature neurons prevents effective DNA transfection using simple, cationic lipid-based methods. Adequate heterologous protein expression is often only achievable using complex techniques that, in many cases, are associated with substantial toxicity. Here, a simple method for high efficiency transfection of mammalian primary neurons using in vitro transcribed mRNA and the cationic lipid transfection reagent Lipofectamine⹠2000 is described. Optimal transfection conditions were established in adult mouse dissociated dorsal root ganglion (DRG) neurons using a 96-well based luciferase activity assay. Using these conditions, a transfection efficiency of 25% was achieved in DRG neurons transfected with EGFP mRNA. High transfection efficiencies were also obtained in dissociated rat superior cervical ganglion (SCG) neurons and mouse cortical and hippocampal cultures. Endogenous Ca2+ currents in EGFP mRNA-transfected SCG neurons were not significantly different from untransfected neurons, which suggested that this technique is well suited for heterologous expression in patch clamp recording experiments. Functional expression of a cannabinoid receptor (CB1R), a G protein inwardly rectifying K+ channel (GIRK4) and a dominant-negative G protein α-subunit mutant (GoA G203T) indicate that the levels of heterologous protein expression attainable using mRNA transfection are suitable for most functional protein studies. This study demonstrates that mRNA transfection is a straightforward and effective method for heterologous expression in neurons and is likely to have many applications in neuroscience research
Rapid Modification of Proteins Using a Rapamycin-Inducible Tobacco Etch Virus Protease System
The ability to disrupt the function of a specific protein on a rapid time scale provides a powerful tool for biomedical research. Specific proteases provide a potential method to selectively cleave a chosen protein, but rapid control of protease activity is difficult.A heterologous expression system for rapid target-directed proteolysis in mammalian cells was developed. The system consists of an inducible NIa protease from the tobacco etch virus (TEVp) and a chosen protein into which a TEVp substrate recognition sequence (TRS) has been inserted. Inducible activity was conferred to the TEVp using rapamycin-controlled TEVp fragment complementation. TEVp activity was assayed using a FRET-based reporter construct. TEVp expression was well tolerated by mammalian cells and complete cleavage of the substrate was possible. Cleavage at 37 degrees C proceeded exponentially with a time constant of approximately 150 minutes. Attempts to improve cleavage efficiency were hampered by substantial background activity, which was attributed to inherent affinity between the TEVp fragments. A second TEVp assay, based on changes in inactivation of a modified K(V)3.4 channel, showed that functional properties of a channel can be using altered using an inducible TEVp system. Similar levels of background activity and variability were observed in both electrophysiological and FRET assays.The results suggested that an optimum level of TEVp expression leading to sufficient inducible activity (with minimal background activity) exists but the variability in expression levels between cells makes the present system rather impractical for single cell experiments. The system is likely to be more suitable for experiments in which the cell-to-cell variability is less of an issue; for example, in experiments involving large populations of cells
Missing the target: including perspectives of women with overweight and obesity to inform stigma-reduction strategies
OBJECTIVE: Pervasive weight stigma and discrimination have led to ongoing calls for efforts to reduce this bias. Despite increasing research on stigma-reduction strategies, perspectives of individuals who have experienced weight stigma have rarely been included to inform this research. The present study conducted a systematic examination of women with high body weight to assess their perspectives about a broad range of strategies to reduce weight-based stigma.
METHODS: Women with overweight or obesity (N = 461) completed an online survey in which they evaluated the importance, feasibility and potential impact of 35 stigma-reduction strategies in diverse settings. Participants (91.5% who reported experiencing weight stigma) also completed self-report measures assessing experienced and internalized weight stigma.
RESULTS: Most participants assigned high importance to all stigma-reduction strategies, with school-based and healthcare approaches accruing the highest ratings. Adding weight stigma to existing anti-harassment workplace training was rated as the most impactful and feasible strategy. The family environment was viewed as an important intervention target, regardless of participants\u27 experienced or internalized stigma.
CONCLUSION: These findings underscore the importance of including people with stigmatized identities in stigma-reduction research; their insights provide a necessary and valuable contribution that can inform ways to reduce weight-based inequities and prioritize such efforts
Free Thermal Convection Driven by Nonlocal Effects
We report and explain a convective phenomenon observed in molecular dynamics
simulations that cannot be classified either as a hydrodynamics instability nor
as a macroscopically forced convection. Two complementary arguments show that
the velocity field by a thermalizing wall is proportional to the ratio between
the heat flux and the pressure. This prediction is quantitatively corroborated
by our simulations.Comment: RevTex, figures is eps, submited for publicatio
Plasma convection in the magnetotail lobes: statistical results from Cluster EDI measurements
A major part of the plasma in the Earth's magnetotail is populated through transport of plasma from the solar wind via the magnetotail lobes. In this paper, we present a statistical study of plasma convection in the lobes for different directions of the interplanetary magnetic field and for different geomagnetic disturbance levels. The data set used in this study consists of roughly 340 000 one-minute vector measurements of the plasma convection from the Cluster Electron Drift Instrument (EDI) obtained during the period February 2001 to June 2007. The results show that both convection magnitude and direction are largely controlled by the interplanetary magnetic field (IMF). For a southward IMF, there is a strong convection towards the central plasma sheet with convection velocities around 10 km s<sup>&minus;1</sup>. During periods of northward IMF, the lobe convection is almost stagnant. A <I>B<sub>y</sub></I> dominated IMF causes a rotation of the convection patterns in the tail with an oppositely directed dawn-dusk component of the convection for the northern and southern lobe. Our results also show that there is an overall persistent duskward component, which is most likely a result of conductivity gradients in the footpoints of the magnetic field lines in the ionosphere
Thermal convection in fluidized granular systems
Thermal convection is observed in molecular dynamic simulation of a fluidized
granular system of nearly elastic hard disks moving under gravity, inside a
rectangular box. Boundaries introduce no shearing or time dependence, but the
energy injection comes from a slip (shear-free) thermalizing base. The top wall
is perfectly elastic and lateral boundaries are either elastic or periodic. The
observed convection comes from the effect of gravity and the spontaneous
granular temperature gradient that the system dynamically develops.Comment: 4 pages, 5 figure
Do Health Beliefs and Behaviors Differ According to Severity of Obesity? A Qualitative Study of Australian Adults
Public responses to obesity have focused on providing standardized messages and supports to all obese individuals, but there is limited understanding of the impact of these messages on obese adults. This descriptive qualitative study using in-depth interviews and a thematic method of analysis, compares the health beliefs and behaviors of 141 Australian adults with mild to moderate (BMI 30â39.9) and severe (BMI â„ 40) obesity. Mildly obese individuals felt little need to change their health behaviors or to lose weight for health reasons. Most believed they could âlose weightâ if they needed to, distanced themselves from the word obesity, and stigmatized those âfatterâ than themselves. Severely obese individuals felt an urgent need to change their health behaviors, but felt powerless to do so. They blamed themselves for their weight, used stereotypical language to describe their health behaviors, and described being âat warâ with their bodies. Further research, particularly about the role of stigma and stereotyping, is needed to fully understand the impact of obesity messaging on the health beliefs, behaviors, and wellbeing of obese and severely obese adults
Responses to gestational weight management guidance: a thematic analysis of comments made by women in online parenting forums
Background: The National Institute for Health and Clinical Excellence (NICE) published guidance on weight management in pregnancy in July 2010[1], and this received considerable press coverage across a range of media. This offered an opportunity to examine how gestational weight management guidance was received by UK women.
Methods: A thematic analysis was conducted of 400 posts made in UK-based parenting internet forums in the week following the publication of the NICE guidance. This allowed us to examine the naturally occurring comments from 202 women who posted about the guidance on public forums.
Results: Three main themes were identified and explored: i) Perceived control/responsibility ii) Risk perception iii) Confused messages.
Conclusions: Women differed in their perceptions of the level of control that they had over being overweight with some feeling responsible and motivated to maintain a healthy lifestyle. Others felt there were multiple factors influencing their weight issues beyond their control. There were reports of feeling guilty about the impact of weight on the growing baby and experiencing significant obesity stigma from the public and health professionals. Information about the risks of overweight and obesity in pregnancy were difficult messages for women to hear, and for health professionals to deliver. Women reported being confused by the messages that they received. Health messages need to be delivered sensitively to women, and health professionals need support and training to do this. Risk information should always be accompanied with clear advice and support to help women to manage their weight in pregnancy.
Keywords: internet-mediated research, gestational weight gain, parenting forums, NICE, women, views, risk perception</p
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