Analysis of polyubiquitin conjugates reveals that the Rpn10 substrate receptor contributes to the turnover of multiple proteasome targets

The polyubiquitin receptor Rpn10 targets ubiquitylated Sic1 to the 26S proteasome for degradation. In contrast, turnover of at least one ubiquitin-proteasome system (UPS) substrate, CPY*, is impervious to deletion of RPN10. To distinguish whether RPN10 is involved in the turnover of only a small set of cell cycle regulators that includes Sic1 or plays a more general role in the UPS, we sought to develop a general method that would allow us to survey the spectrum of ubiquitylated proteins that selectively accumulate in rpn10 cells. Polyubiquitin conjugates from yeast cells that express hexahistidine-tagged ubiquitin (H6-ubiquitin) were first enriched on a polyubiquitin binding protein affinity resin. This material was then denatured and subjected to IMAC to retrieve H6-ubiquitin and proteins to which it may be covalently linked. Using this approach, we identified 127 proteins that are candidate substrates for the 26S proteasome. We then sequenced ubiquitin conjugates from cells lacking Rpn10 (rpn10) and identified 54 proteins that were uniquely recovered from rpn10 cells. These include two known targets of the UPS, the cell cycle regulator Sic1 and the transcriptional activator Gcn4. Our approach of comparing the ubiquitin conjugate proteome in wild-type and mutant cells has the resolving power to identify even an extremely inabundant transcriptional regulatory protein and should be generally applicable to mapping enzyme substrate networks in the UPS

Ray-optical refraction with confocal lenslet arrays

Two parallel lenslet arrays with focal lengths f1 and f2 that share a common focal plane (that is, which are separated by a distance f1+f2) can refract transmitted light rays according to Snell's law, but with the 'sin's replaced with 'tan's. This is the case for a limited range of input angles and other conditions. Such confocal lenslet arrays can therefore simulate the interface between optical media with different refractive indices, n1 and n2, whereby the ratio η=-f2/f1 plays the role of the refractive-index ratio n2/n1. Suitable choices of focal lengths enable positive and negative refraction. In contrast to Snell's law, which leads to nontrivial geometric imaging by a planar refractive-index interface only for the special case of n1=±n2, the modified refraction law leads to geometric imaging by planar confocal lenslet arrays for any value of η. We illustrate some of the properties of confocal lenslet arrays with images rendered using ray-tracing software

Extinction of cue-evoked food seeking recruits a GABAergic interneuron ensemble in the dorsal medial prefrontal cortex of mice

Animals must quickly adapt food-seeking strategies to locate nutrient sources in dynamically changing environments. Learned associations between food and environmental cues that predict its availability promote food-seeking behaviors. However, when such cues cease to predict food availability, animals undergo 'extinction' learning, resulting in the inhibition of food-seeking responses. Repeatedly activated sets of neurons, or 'neuronal ensembles', in the dorsal medial prefrontal cortex (dmPFC) are recruited following appetitive conditioning and undergo physiological adaptations thought to encode cue-reward associations. However, little is known about how the recruitment and intrinsic excitability of such dmPFC ensembles are modulated by extinction learning. Here, we used in vivo 2-Photon imaging in male Fos-GFP mice that express green fluorescent protein (GFP) in recently behaviorally-activated neurons to determine the recruitment of activated pyramidal and GABAergic interneuron mPFC ensembles during extinction. During extinction, we revealed a persistent activation of a subset of interneurons which emerged from a wider population of interneurons activated during the initial extinction session. This activation pattern was not observed in pyramidal cells, and extinction learning did not modulate the excitability properties of activated neurons. Moreover, extinction learning reduced the likelihood of reactivation of pyramidal cells activated during the initial extinction session. Our findings illuminate novel neuronal activation patterns in the dmPFC underlying extinction of food-seeking, and in particular, highlight an important role for interneuron ensembles in this inhibitory form of learning

Is diagnostic tonsillectomy indicated in all children with asymmetrically enlarged tonsil?

Objectives. The aims of the study were: (i) to determine the necessity for diagnostic tonsillectomy in children with asymmetrically enlarged tonsils; (ii) to determine the accuracy of clinical assessment of tonsillar asymmetry; and (iii) to determine how to manage children with clinical tonsillar asymmetry in a developing-world practice. Methods. A prospective study was carried out at Red Cross War Memorial Children’s Hospital in Cape Town, over an 8-month period. All children undergoing tonsillectomy or adenotonsillectomy had a clinical assessment of tonsil symmetry done, and all tonsil and adenoid specimens were examined histologically. The maximum diameter and volume of the resected tonsils were measured. A comparison was done of true tonsil asymmetry in patients with asymmetrical tonsils and a subgroup of matched controls with symmetrical tonsils. Results. A total of 344 tonsils were analysed (172 patients). The 13 patients (7.6%) diagnosed as having clinically asymmetrically enlarged tonsils had no significant pathological diagnosis. In the patients with symmetrical tonsils there were abnormal pathological findings (tuberculosis of the adenoids and T-cell lymphoma of the tonsils and adenoids). In the clinically asymmetrical tonsil group, true tonsillar asymmetry was 3 mm (maximum diameter), and 2.2 cm3 (volume), compared with 1.9 mm and 1.5 cm3 in the symmetrical tonsil group. When patients with clinical tonsillar asymmetry and symmetry were compared, the difference in maximum diameter (p = 0.62) and volume (p = 0.73) was not significantly different. Conclusions. Clinical tonsillar asymmetry is usually apparent rather than real. The incidence of significant pathology in children with asymptomatic, asymmetrical tonsils is low. Diagnostic tonsillectomy is indicated in children with asymmetrically enlarged tonsils associated with constitutional symptoms, cervical lymphadenopathy, rapid tonsil enlargement or significant tonsillar asymmetry

High Gaussicity feedhorns for sub-/ millimeter wave applications

In feedhorn design, the power coupling to the fundamental free-space LG00 mode, or Gaussicity, is a good proxy for high performance, particularly the sidelobe and cross-polar levels and the near-field behavior. Gaussicity can be maximized by ensuring that the first few horn modes reach the aperture with the appropriate phase and amplitude relationship. We present two feedhorn designs for which the Gaussicity was maximized in order to achieve high performance. The first is a 94 GHz corrugated horn with a tanh-linear profile, manufactured by electroforming, which achieves a Gaussicity of 99.92% at band center and sidelobes at the -60 dB level. The second is a 340 GHz smooth-walled spline horn which achieves a Gaussicity of >99.2% over a 10% bandwidth, sidelobes below -30 dB and excellent near-field behavior. This design has been successfully fabricated in E-plane split block suitable for low volume manufacture, for example for imaging arrays.Postprin

The Stokes-Einstein-Sutherland equation at the nanoscale revisited

The Stokes-Einstein-Sutherland (SES) equation is at the foundation of statistical physics, relating a particle's diffusion coefficient and size with the fluid viscosity, temperature and the boundary condition for the particle-solvent interface. It is assumed that it relies on the separation of scales between the particle and the solvent, hence it is expected to break down for diffusive transport on the molecular scale. However, a number of experimental studies showed a remarkable small, if any, violation of this equation down to the size of a nm, where there is no scale separation. To resolve this puzzle we combine analytical ultracentrifugation experiments and molecular dynamics simulations to study the transport of buckminsterfullerene C$_{60}$ suspended in toluene at infinite dilution. We show that this system clearly violates the conditions of slow momentum relaxation. Yet, through a linear response to a constant force, we show both in experiments and in simulations that the SES equation can be recovered in the long time limit with no more than 4% uncertainty. This nonetheless requires partial slip on the particle interface, extracted consistently from all the data. Our results, thus, resolve a long-standing discussion on the validity and limits of the SES equation at the molecular scale

The secretion inhibitor Exo2 perturbs trafficking of Shiga toxin between endosomes and the trans-Golgi network

The small-molecule inhibitor Exo2 {4-hydroxy-3-methoxy-(5,6,7,8-tetrahydrol[1]benzothieno[2,3-d]pyrimidin-4-yl)hydraz-one benzaldehyde} has been reported to disrupt the Golgi apparatus completely and to stimulate Golgi–ER (endoplasmic reticulum) fusion in mammalian cells, akin to the well-characterized fungal toxin BFA (brefeldin A). It has also been reported that Exo2 does not affect the integrity of the TGN (trans-Golgi network), or the direct retrograde trafficking of the glycolipid-binding cholera toxin from the TGN to the ER lumen. We have examined the effects of BFA and Exo2, and found that both compounds are indistinguishable in their inhibition of anterograde transport and that both reagents significantly disrupt the morphology of the TGN in HeLa and in BS-C-1 cells. However, Exo2, unlike BFA, does not induce tubulation and merging of the TGN and endosomal compartments. Furthermore, and in contrast with its effects on cholera toxin, Exo2 significantly perturbs the delivery of Shiga toxin to the ER. Together, these results suggest that the likely target(s) of Exo2 operate at the level of the TGN, the Golgi and a subset of early endosomes, and thus Exo2 provides a more selective tool than BFA for examining membrane trafficking in mammalian cells

Local light-ray rotation

We present a sheet structure that rotates the local ray direction through an arbitrary angle around the sheet normal. The sheet structure consists of two parallel Dove-prism sheets, each of which flips one component of the local direction of transmitted light rays. Together, the two sheets rotate transmitted light rays around the sheet normal. We show that the direction under which a point light source is seen is given by a Mobius transform. We illustrate some of the properties with movies calculated by ray-tracing software.Comment: 9 pages, 6 figure

A solvable model of a random spin-1/2 XY chain

The paper presents exact calculations of thermodynamic quantities for the spin-1/2 isotropic XY chain with random lorentzian intersite interaction and transverse field that depends linearly on the surrounding intersite interactions.Comment: 14 pages (Latex), 2 tables, 13 ps-figures included, (accepted for publication in Phys.Rev.B