A pre-embedding immunogold approach for detection of synaptic endocytic proteins in situ

During the past decade, many molecular components of clathrin-mediated endocytosis have been identified and proposed to play various hypothetical roles in the process [Nat. Rev. Neurosci. 1 (2000) 161; Nature 422 (2003) 37]. One limitation to the evaluation of these hypotheses is the efficiency and resolution of immunolocalization protocols currently in use. In order to facilitate the evaluation of these hypotheses and to understand more fully the molecular mechanisms of clathrin-mediated endocytosis, we have developed a protocol allowing enhanced and reliable subcellular immunolocalization of proteins in synaptic endocytic zones in situ. Synapses established by giant reticulospinal axons in lamprey are used as a model system for these experiments. These axons are unbranched and reach up to 80-100 microm in diameter. Synaptic active zones and surrounding endocytic zones are established on the surface of the axonal cylinder. To provide access for antibodies to the sites of synaptic vesicle recycling, axons are lightly fixed and cut along their longitudinal axis. To preserve the ultrastructure of the synaptic endocytic zone, antibodies are applied without the addition of detergents. Opened axons are incubated with primary antibodies, which are detected with secondary antibodies conjugated to gold particles. Specimens are then post-fixed and processed for electron microscopy. This approach allows preservation of the ultrastructure of the endocytic sites during immunolabeling procedures, while simultaneously achieving reliable immunogold detection of proteins on endocytic intermediates. To explore the utility of this approach, we have investigated the localization of a GTPase, dynamin, on clathrin-coated intermediates in the endocytic zone of the lamprey giant synapse. Using the present immunogold protocol, we confirm the presence of dynamin on late stage coated pits [Nature 422 (2003) 37] and also demonstrate that dynamin is recruited to the coat of endocytic intermediates from the very early stages of the clathrin coat formation. Thus, our experiments show that the current pre-embedding immunogold method is a useful experimental tool to study the molecular mechanisms of synaptic vesicle recycling.

Pre-Radiotherapy Dental Assessment of Head and Neck Oncology Patients: An Audit.

Objectives: Approximately 300 people are diagnosed with Head and Neck cancer annually in Northern Ireland. The management may include treatment by surgery or by chemotherapy and radiotherapy,
or a combination of modalities. Patients whose oral cavity, teeth, salivary glands and jaws that
will be affected by treatment, particularly radiotherapy should have a pre-treatment assessment. This should be done as early as possible to maximise the time available for dental management. However, this can be challenging owing to the complexities of cancer diagnosis, treatment planning and multidisciplinary management. At the Belfast Dental Hospital, a number of patients were referred post- radiotherapy with complications after not having received a pre-treatment assessment. The referrals for pre- treatment dental assessment were also late in patients’ multidisciplinary journey, limiting the time period
for dental input. The purpose of this audit was to examine the time period between dental assessment and commencement of radiotherapy and whether this was an adequate time frame for dental management. This audit will also examine the dental diseases present and the treatments required pre-radiotherapy. Methods: Data for this audit was collected over 4 months in 2012
by analysing the dental charts and referrals of new patients who were referred to and attended the dental head and neck oncology clinic. A standardised referral pro-forma was introduced from September 2013 to improve the referral process.
A re-audit was conducted over 4 months in 2014. Data was collected similarly as previous. The time period between dental assessment and commencement of radiotherapy was examined. The presence of dental disease and subsequent treatments required were also noted.
Results: 63 new patients were examined in the dental head and neck oncology clinic over 4 months in 2012. 48 (76.2%) were examined pre-radiotherapy. The average length of time between dental assessment and radiotherapy commencement was 11 days. A new standardised referral pro-forma was introduced in 2013. In the re-audit, 65 new patients were seen over 4 months in 2014.
60 (92.3%) patients were examined pre-radiotherapy. The average length of time between dental assessment and radiotherapy commencement was 18 days.
Conclusion: Given the high prevalence of pre-existing dental disease amongst head and neck cancer patients, prompt dental assessment and treatment is vital. Efforts aimed at improving the care pathway are on-going through the implementation of a mandatory referral pro-forma and a dedicated assessment clinic.

Self-regulated Shocks in Massive Star Binary Systems

In an early-type, massive star binary system, X-ray bright shocks result from the powerful collision of stellar winds driven by radiation pressure on spectral line transitions. We examine the influence of the X-rays from the wind-wind collision shocks on the radiative driving of the stellar winds using steady-state models that include a parameterized line force with X-ray ionization dependence. Our primary result is that X-ray radiation from the shocks inhibits wind acceleration and can lead to a lower pre-shock velocity, and a correspondingly lower shocked plasma temperature, yet the intrinsic X-ray luminosity of the shocks, L X, remains largely unaltered, with the exception of a modest increase at small binary separations. Due to the feedback loop between the ionizing X-rays from the shocks and the wind driving, we term this scenario as self-regulated shocks. This effect is found to greatly increase the range of binary separations at which a wind-photosphere collision is likely to occur in systems where the momenta of the two winds are significantly different. Furthermore, the excessive levels of X-ray ionization close to the shocks completely suppress the line force, and we suggest that this may render radiative braking less effective. Comparisons of model results against observations reveal reasonable agreement in terms of log (L X/L bol). The inclusion of self-regulated shocks improves the match for kT values in roughly equal wind momenta systems, but there is a systematic offset for systems with unequal wind momenta (if considered to be a wind-photosphere collision).

Pre-Processing Power Traces with a Phase-Sensitive Detector

As cryptographic implementations are increasingly subsumed as functional blocks within larger systems on chip, it becomes more difficult to identify the power consumption signatures of cryptographic operations amongst other unrelated processing activities. In addition, at higher clock frequencies, the current decay between successive processing rounds is only partial, making it more difficult to apply existing pattern matching techniques in side-channel analysis. We show however, through the use of a phase-sensitive detector, that power traces can be pre-processed to generate a filtered output which exhibits an enhanced round pattern, enabling the identification of locations on a device where encryption operations are occurring and also assisting with the re-alignment of power traces for side-channel attacks.

High Performance Real-time Pre-Processing for Fixed-Complexity Sphere Decoder

Pre-processing (PP) of received symbol vector and channel matrices is an essential pre-requisite operation for Sphere Decoder (SD)-based detection of Multiple-Input Multiple-Output (MIMO) wireless systems. PP is a highly complex operation, but relative to the total SD workload it represents a relatively small fraction of the overall computational cost of detecting an OFDM MIMO frame in standards such as 802.11n. Despite this, real-time PP architectures are highly inefficient, dominating the resource cost of real-time SD architectures. This paper resolves this issue. By reorganising the ordering and QR decomposition sub operations of PP, we describe a Field Programmable Gate Array (FPGA)-based PP architecture for the Fixed Complexity Sphere Decoder (FSD) applied to 4 × 4 802.11n MIMO which reduces resource cost by 50% as compared to state-of-the-art solutions whilst maintaining real-time performance.

A population-based study of visual impairment among pre-school children in Beijing: the Beijing study of visual impairment in children.

PURPOSE: To evaluate the prevalence and causes of visual impairment among Chinese children aged 3 to 6 years in Beijing. DESIGN: Population-based prevalence survey. METHODS: Presenting and pinhole visual acuity were tested using picture optotypes or, in children with pinhole vision < 6/18, a Snellen tumbling E chart. Comprehensive eye examinations and cycloplegic refraction were carried out for children with pinhole vision < 6/18 in the better-seeing eye. RESULTS: All examinations were completed on 17,699 children aged 3 to 6 years (95.3% of sample). Subjects with bilateral correctable low vision (presenting vision < 6/18 correctable to >or= 6/18) numbered 57 (0.322%; 95% confidence interval [CI], 0.237% to 0.403%), while 14 (0.079%; 95% CI, 0.038% to 0.120%) had bilateral uncorrectable low vision (best-corrected vision of < 6/18 and >or= 3/60), and 5 subjects (0.028%; 95% CI, 0.004% to 0.054%) were bilaterally blind (best-corrected acuity < 3/60). The etiology of 76 cases of visual impairment included: refractive error in 57 children (75%), hereditary factors (microphthalmos, congenital cataract, congenital motor nystagmus, albinism, and optic nerve disease) in 13 children (17.1 %), amblyopia in 3 children (3.95%), and cortical blindness in 1 child (1.3%). The cause of visual impairment could not be established in 2 (2.63%) children. The prevalence of visual impairment did not differ by gender, but correctable low vision was significantly (P < .0001) more common among urban as compared with rural children. CONCLUSION: The leading causes of visual impairment among Chinese preschool-aged children are refractive error and hereditary eye diseases. A higher prevalence of refractive error is already present among urban as compared with rural children in this preschool population.

Distant activity of 67P/Churyumov-Gerasimenko in 2014: Ground-based results during the Rosetta pre-landing phase

As the ESA Rosetta mission approached, orbited, and sent a lander to comet 67P/Churyumov-Gerasimenko in 2014, a large campaign of ground-based observations also followed the comet. We constrain the total activity level of the comet by photometry and spectroscopy to place Rosetta results in context and to understand the large-scale structure of the comet's coma pre-perihelion. We performed observations using a number of telescopes, but concentrate on results from the 8m VLT and Gemini South telescopes in Chile. We use R-band imaging to measure the dust coma contribution to the comet's brightness and UV-visible spectroscopy to search for gas emissions, primarily using VLT/FORS. In addition we imaged the comet in near-infrared wavelengths (JHK) in late 2014 with Gemini-S/Flamingos 2. We find that the comet was already active in early 2014 at heliocentric distances beyond 4 au. The evolution of the total activity (measured by dust) followed previous predictions. No gas emissions were detected despite sensitive searches. The comet maintains a similar level of activity from orbit to orbit, and is in that sense predictable, meaning that Rosetta results correspond to typical behaviour for this comet. The gas production (for CN at least) is highly asymmetric with respect to perihelion, as our upper limits are below the measured production rates for similar distances post-perihelion in previous orbits.