FM1-43 dye behaves as a permeant blocker of the hair-cell mechanotransducer channel

Hair cells in mouse cochlear cultures are selectively labeled by brief exposure to FM1-43, a styryl dye used to study endocytosis and exocytosis. Real-time confocal microscopy indicates that dye entry is rapid and via the apical surface. Cooling to 4°C and high extracellular calcium both reduce dye loading. Pretreatment with EGTA, a condition that breaks tip links and prevents mechanotransducer channel gating, abolishes subsequent dye loading in the presence of calcium. Dye loading recovers after calcium chelation with a time course similar to that described for tip-link regeneration. Myo7a mutant hair cells, which can transduce but have all mechanotransducer channels normally closed at rest, do not label with FM1-43 unless the bundles are stimulated by large excitatory stimuli. Extracellular perfusion of FM1-43 reversibly blocks mechanotransduction with half-blocking concentrations in the low micromolar range. The block is reduced by high extracellular calcium and is voltage dependent, decreasing at extreme positive and negative potentials, indicating that FM1-43 behaves as a permeant blocker of the mechanotransducer channel. The time course for the relief of block after voltage steps to extreme potentials further suggests that FM1-43 competes with other cations for binding sites within the pore of the channel. FM1-43 does not block the transducer channel from the intracellular side at concentrations that would cause complete block when applied extracellularly. Calcium chelation and FM1-43 both reduce the ototoxic effects of the aminoglycoside antibiotic neomycin sulfate, suggesting that FM1-43 and aminoglycosides enter hair cells via the same pathway

158µJ pulses from a single transverse mode, large mode-area EDFA

We report the amplification of 10pJ-100pJ, semiconductor diode pulses up to an energy of 158µJ and peak powers >100kW in a multi-stage fibre amplifier chain based on a single-mode, large mode-area erbium doped amplifier design. These results represent the highest single-mode pulse energy ever extracted from any doped fibre system

Dependence of the spike-triggered average voltage on membrane response properties

The spike-triggered average voltage (STV) is an experimentally measurable quantity that is determined by both the membrane response properties and the statistics of the synaptic drive. Here, the form of the STV is modelled for neurons with three distinct types of subthreshold dynamics; passive decay, h-current sag, and damped oscillations. Analytical expressions for the STV are first obtained in the low-noise limit, identifying how the subthreshold dynamics of the cell affect its form. A second result is then derived that captures the power-law behaviour of the STV near the spike threshold

Generators of simple Lie algebras in arbitrary characteristics

In this paper we study the minimal number of generators for simple Lie algebras in characteristic 0 or p > 3. We show that any such algebra can be generated by 2 elements. We also examine the 'one and a half generation' property, i.e. when every non-zero element can be completed to a generating pair. We show that classical simple algebras have this property, and that the only simple Cartan type algebras of type W which have this property are the Zassenhaus algebras.Comment: 26 pages, final version, to appear in Math. Z. Main improvements and corrections in Section 4.

Improving College Access and Completion for Low-Income and First-Generation Students: The Role of College Access and Success Programs

Resequencing of Agaricus bisporus herbarium specimen 99817338

How to Open the Treasure Chest? Optimising DNA Extraction from Herbarium Specimens

Herbarium collections are potentially an enormous resource for DNA studies, but the use of herbarium specimens in molecular studies has thus far been slowed down by difficulty in obtaining amplifiable DNA. Here we compare a set of commercially available DNA extraction protocols and their performance in terms of DNA purity and yield, and PCR amplification success as measured by using three differentially sized markers, the rbcL barcoding marker (cpDNA), the LEAFY exon 3 (nrDNA), and the trnL(UAA) P6 loop (cpDNA). Results reveal large differences between extraction methods, where DNA purity rather than yield is shown to be strongly correlated with PCR success. Amplicon size shows similarly strong correlation with PCR success, with the shortest fragment showing the highest success rate (78%, P6 loop, 10–143 base pairs (bp)) and the largest fragment the lowest success (10%, rbcL, 670 bp). The effect of specimen preparation method on PCR success was also tested. Results show that drying method strongly affects PCR success, especially the availability of fragments longer than 250 bp, where longer fragments are more available for PCR amplification in air dried material compared to alcohol dried specimens. Results from our study indicate that projects relying on poor-quality starting material such as herbarium or scat samples should focus on extracting pure DNA and aim to amplify short target regions

Accessing elite nurses for research: reflections on the theoretical and practical issues of telephone interviewing

Elite groups are interesting as they frequently are powerful (in terms of position, knowledge and influence) and enjoy considerable authority. It is important, therefore, to involve them in research concerned with understanding social contexts and processes. This is particularly pertinent in healthcare, where considerable strategic development and change are features of everyday practice that may be guided or perceived as being guided, by elites. This paper evolved from a study investigating the availability and role of nurses whose remit involved leading nursing research and development within acute NHS Trusts in two health regions in Southern England. The study design included telephone interviews with Directors of Nursing Services during which time the researchers engaged in a reflective analysis of their experiences of conducting research with an `elite' group. Important issues identified were the role of gatekeepers, engagement with elites and the use of the telephone interview method in this context. The paper examines these issues and makes a case for involving executive nurses in further research. The paper also offers strategies to help researchers design and implement telephone interview studies successfully to maximise access to the views and experiences of `hard to reach groups', such as elites, while minimising the associated disruption

Estimating Associations Between Annual Concentrations of Particulate Matter and Mortality in the United States, Using Data Linkage and Bayesian Maximum Entropy

Background: Exposure to fine particulate matter (PM2.5) is an established risk factor for human mortality. However, previous US studies have been limited to select cities or regions or to population subsets (e.g., older adults). Methods: Here, we demonstrate how to use the novel geostatistical method Bayesian maximum entropy to obtain estimates of PM2.5 concentrations in all contiguous US counties, 2000–2016. We then demonstrate how one could use these estimates in a traditional epidemiologic analysis examining the association between PM2.5 and rates of all-cause, cardiovascular, respiratory, and (as a negative control outcome) accidental mortality. Results: We estimated that, for a 1 log(μg/m3) increase in PM2.5 concentration, the conditional all-cause mortality incidence rate ratio (IRR) was 1.029 (95% confidence interval [CI]: 1.006, 1.053). This implies that the rate of all-cause mortality at 10 µg/m3 would be 1.020 times the rate at 5 µg/m3. IRRs were larger for cardiovascular mortality than for all-cause mortality in all gender and race–ethnicity groups. We observed larger IRRs for all-cause, nonaccidental, and respiratory mortality in Black non-Hispanic Americans than White non-Hispanic Americans. However, our negative control analysis indicated the possibility for unmeasured confounding. Conclusion: We used a novel method that allowed us to estimate PM2.5 concentrations in all contiguous US counties and obtained estimates of the association between PM2.5 and mortality comparable to previous studies. Our analysis provides one example of how Bayesian maximum entropy could be used in epidemiologic analyses; future work could explore other ways to use this approach to inform important public health questions