5 '-Untranslated regions with multiple upstream AUG codons can support low-level translation via leaky scanning and reinitiation

Upstream AUGs (uAUGs) and upstream open reading frames (uORFs) are common features of mRNAs that encode regulatory proteins and have been shown to profoundly influence translation of the main ORF. In this study, we employed a series of artificial 5'-untranslated regions (5'-UTRs) containing one or more uAUGs/uORFs to systematically assess translation initiation at the main AUG by leaky scanning and reinitiation mechanisms. Constructs containing either one or two uAUGs in varying contexts but without an in-frame stop codon upstream of the main AUG were used to analyse the leaky scanning mechanism. This analysis largely confirmed the ranking of different AUG contextual sequences that was determined previously by Kozak. In addition, this ranking was the same for both the first and second uAUGs, although the magnitude of initiation efficiency differed. Moreover, similar to10% of ribosomes exhibited leaky scanning at uAUGs in the most favourable context and initiated at a downstream AUG. A second group of constructs containing different numbers of uORFs, each with optimal uAUGs, were used to measure the capacity for reinitiation. We found significant levels of initiation at the main ORF even in constructs containing four uORFs, with nearly 10% of ribosomes capable of reinitiating five times. This study shows that for mRNAs containing multiple uORFs/uAUGs, ribosome reinitiation and leaky scanning are efficient mechanisms for initiation at their main AUGs.

Psilocybin impairs high-level but not low-level motion perception

The hallucinogenic serotonin(IA&2A) agonist psilocybin is known for its ability to induce illusions of motion in otherwise stationary objects or textured surfaces. This study investigated the effect of psilocybin on local and global motion processing in nine human volunteers. Using a forced choice direction of motion discrimination task we show that psilocybin selectively impairs coherence sensitivity for random dot patterns, likely mediated by high-level global motion detectors, but not contrast sensitivity for drifting gratings, believed to be mediated by low-level detectors. These results are in line with those observed within schizophrenic populations and are discussed in respect to the proposition that psilocybin may provide a model to investigate clinical psychosis and the pharmacological underpinnings of visual perception in normal populations.

On the optimal ratio of heavy to light chain genes for efficient recombinant antibody production by CHO cells

Monoclonal antibodies (Mab) are heterotetramers consisting of an equimolar ratio of heavy chain (HC) and light chain (LC) polypeptides. Accordingly, most recombinant Mab expression systems utilize an equimolar ratio of heavy chain (he) to light chain (lc) genes encoded on either one or two plasmids. However, there is no evidence to suggest that this gene ratio is optimal for stable or transient production of recombinant Mab. In this study we have determined the optimal ratio of hc:lc genes for production of a recombinant IgG(4) Mab, cB72.3, by Chinese hamster ovary (CHO) cells using both empirical and mathematical modeling approaches. Polyethyleneimine-mediated transient expression of cB72.3 at varying ratios of hc:lc genes encoded on separate plasmids yielded an optimal Mab titer at a hc:lc gene ratio of 3:2; a conclusion confirmed by separate mathematical modeling of the Mab folding and assembly process using transient expression data. On the basis of this information, we hypothesized that utilization of he genes at low hc:lc gene ratios is more efficient. To confirm this, cB72.3 Mab was transiently produced by CHO cells at constant he and varying lc gene dose. Under these conditions, Mab yield was increased with a concomitant increase in lc gene dose. To determine if the above findings also apply to stably transfected CHO cells producing recombinant Mab, we compared the intra- and extracellular ratios of HC and LC polypeptides for three GS-CHO cells lines transfected with a 1:1 ratio of hc:lc genes and selected for stable expression of the same recombinant Mab, cB72.3. Intra- and extracellular HC:LC polypeptide ratios ranged from 1:2 to 1:5, less than that observed on transient expression of the same Mab in parental CHO cells using the same vector. In conclusion, our data suggest that the optimal ratio of hc:lc genes used for transient and stable expression of Mab differ. In the case of the latter, we infer that optimal Mab production by stably transfected cells represents a compromise between HC abundance limiting productivity and the requirement for excess LC to render Mab folding and assembly more efficient.

Nociceptive scores and endorphin-containing cells reduced by low-level laser therapy (LLLT) in inflamed paws of Wistar rat

Objective: This study aimed to investigate how local pain relief is mediated by laser therapy and how dose affects the relationship. Methods: Inflammation was induced in the hind-paws of Wistar rats. Two groups of rats received 780-nm laser therapy (Spectra-Medics Pty Ltd.) at one of two doses (2.5 and 1 J/cm(2)). One group acted as a control. Scores of nociceptive threshold were recorded using paw pressure and paw thermal threshold measures. Results: A dose of 1 J/cm(2) had no statistically significant effect on antinociceptive responses. A dose of 2.5 J/cm(2) demonstrated a statistically significant effect on paw pressure threshold (p < 0.029) compared to controls. There was no difference in paw thermal threshold responses and paw volumes at either dose. Immunohistochemistry in control animals demonstrated normal beta-endorphin containing lymphocytes in control inflamed paws but no beta-endorphin containing lymphocytes in rats that received laser at 2.5 J/cm(2). Conclusion: The results confirm previous findings that the effect of laser therapy is dose-related. The mechanism of effect may occur via a differentiated pressure-sensitive neural pathway rather than a thermal-sensitive neural pathway. The significance of the immunohistochemistry findings remains unknown.

Low level microsatellite instability may be associated with reduced cancer specific survival in sporadic stage C colorectal carcinoma

Background: Colorectal cancers (CRCs) may be categorised according to the degree of microsatellite instability (MSI) exhibited, as MSI-high (MSI-H), MSI-low (MSI-L), or microsatellite stable (MSS). MSI-H status confers a survival advantage to patients with sporadic CRC. Aims: To determine if low levels of MSI are related to the clinicopathological features and prognosis of sporadic stage C CRC. Patients: A total of 255 patients who underwent resection for sporadic stage C CRC were studied. No patient received chemotherapy. Minimum follow up was five years. Methods: DNA extracted from archival malignant and non-malignant tissue was amplified by polymerase chain reaction using a panel of 11 microsatellites. MSI-H was defined as instability at greater than or equal to40% of markers, MSS as no instability, and MSI-L as instability at >0% but,40% of markers. Patients with MSI-H CRC were excluded from analysis as they have previously been shown to have better survival. Results: Thirty three MSI-L and 176 MSS CRCs were identified. There was no difference in biological characteristics or overall survival of MSI-L compared with MSS CRC but MSI-L was associated with poorer cancer specific survival (hazard ratio 2.0 (95% confidence interval 1.1-3.6)). Conclusions: Sporadic MSI-L and MSS CRCs have comparable clinicopathological features. Further studies are required to assess the impact of MSI-L on prognosis.

Right ventricle extraction by low level and model-based algorithm

In this paper we present an algorithm as the combination of a low level morphological operation and model based Global Circular Shortest Path scheme to explore the segmentation of the Right Ventricle. Traditional morphological operations were employed to obtain the region of interest, and adjust it to generate a mask. The image cropped by the mask is then partitioned into a few overlapping regions. Global Circular Shortest Path algorithm is then applied to extract the contour from each partition. The final step is to re-assemble the partitions to create the whole contour. The technique is deemed quite reliable and robust, as this is illustrated by a very good agreement between the extracted contour and the expert manual drawing output.

Designing a Computer Vision System for Underwater Robotic Interventions

This thesis deals with the challenging problem of designing systems able to perceive objects in underwater environments. In the last few decades research activities in robotics have advanced the state of art regarding intervention capabilities of autonomous systems. State of art in fields such as localization and navigation, real time perception and cognition, safe action and manipulation capabilities, applied to ground environments (both indoor and outdoor) has now reached such a readiness level that it allows high level autonomous operations. On the opposite side, the underwater environment remains a very difficult one for autonomous robots. Water influences the mechanical and electrical design of systems, interferes with sensors by limiting their capabilities, heavily impacts on data transmissions, and generally requires systems with low power consumption in order to enable reasonable mission duration. Interest in underwater applications is driven by needs of exploring and intervening in environments in which human capabilities are very limited. Nowadays, most underwater field operations are carried out by manned or remotely operated vehicles, deployed for explorations and limited intervention missions. Manned vehicles, directly on-board controlled, expose human operators to risks related to the stay in field of the mission, within a hostile environment. Remotely Operated Vehicles (ROV) currently represent the most advanced technology for underwater intervention services available on the market. These vehicles can be remotely operated for long time but they need support from an oceanographic vessel with multiple teams of highly specialized pilots. Vehicles equipped with multiple state-of-art sensors and capable to autonomously plan missions have been deployed in the last ten years and exploited as observers for underwater fauna, seabed, ship wrecks, and so on. On the other hand, underwater operations like object recovery and equipment maintenance are still challenging tasks to be conducted without human supervision since they require object perception and localization with much higher accuracy and robustness, to a degree seldom available in Autonomous Underwater Vehicles (AUV). This thesis reports the study, from design to deployment and evaluation, of a general purpose and configurable platform dedicated to stereo-vision perception in underwater environments. Several aspects related to the peculiar environment characteristics have been taken into account during all stages of system design and evaluation: depth of operation and light conditions, together with water turbidity and external weather, heavily impact on perception capabilities. The vision platform proposed in this work is a modular system comprising off-the-shelf components for both the imaging sensors and the computational unit, linked by a high performance ethernet network bus. The adopted design philosophy aims at achieving high flexibility in terms of feasible perception applications, that should not be as limited as in case of a special-purpose and dedicated hardware. Flexibility is required by the variability of underwater environments, with water conditions ranging from clear to turbid, light backscattering varying with daylight and depth, strong color distortion, and other environmental factors. Furthermore, the proposed modular design ensures an easier maintenance and update of the system over time. Performance of the proposed system, in terms of perception capabilities, has been evaluated in several underwater contexts taking advantage of the opportunity offered by the MARIS national project. Design issues like energy power consumption, heat dissipation and network capabilities have been evaluated in different scenarios. Finally, real-world experiments, conducted in multiple and variable underwater contexts, including open sea waters, have led to the collection of several datasets that have been publicly released to the scientific community. The vision system has been integrated in a state of the art AUV equipped with a robotic arm and gripper, and has been exploited in the robot control loop to successfully perform underwater grasping operations.

Effect of light filters on reading speed in normal and low vision due to age-related macular degeneration

Purpose: To investigate the effects of light filters on reading speed in normal and low vision due to age-related macular degeneration (AMD). Methods: Reading speed was determined for 12 subjects with normal vision and 12 subjects with non-exudative AMD using stationary lowercase nonsensical print in Times Roman font and four light filters; a yellow Corning Photochromic Filter (CPF) 450, a grey neural density (ND) filter, an individual filter obtained using the Intuitive Colorimeter® and a clear filter. Results: There was no statistically significant light filter effect on reading speed for the normal subjects. The AMD group demonstrated a statistically significant 5% average improvement in reading speed with the CPF450 compared with the other filters although some AMD subjects had improvements of 10-15%. Conclusions: Light filters obtained using the Intuitive Colorimeter® performed poorly when compared with the CPF450, ND and clear filters for both the study groups. For the AMD group, average reading speed was statistically greater with the CPF450 than the other filters, however it is questionable whether the improvement (5%) would be clinically significant. As some of the subjects with AMD had greater improvements with the CPF450 we advocate clinical assessment of light filters using existing protocols on an individual basis. © 2004 The College of Optometrists.

Determination of the Pu autoionizing level angular momenta by a multistep excitation with linearly polarized laser light

The modified polarization spectroscopy method was applied for determination of angular momenta of autoionizing states of Pu in multistep resonance ionization processes. In comparison with the known one, our method does not require circular polarization at all, only linear polarizations are needed. This simplicity was reached using a three-dimensional excitation geometry. Angular momenta of nine new autoionizing ²⁴²Pu states were determined. The method suggested could be applied for efficiency improvement in multistep RIMS applications as well as for the odd-even isotope separation for elements with a J = 0 ground state (Pu, Yb, Sm etc.).

A magnetoencephalographic study of low-level auditory processing in developmental dyslexia

The possibility that developmental dyslexia results from low-level sensory processing deficits has received renewed interest in recent years. Opponents of such sensory-based explanations argue that dyslexia arises primarily from phonological impairments. However, many behavioural correlates of dyslexia cannot be explained sufficiently by cognitive-level accounts and there is anatomical, psychometric and physiological evidence of sensory deficits in the dyslexic population. This thesis aims to determine whether the low-level (pre-attentive) processing of simple auditory stimuli is disrupted in compensated adult dyslexics. Using psychometric and neurophysiological measures, the nature of auditory processing abnormalities is investigated. Group comparisons are supported by analysis of individual data in order to address the issue of heterogeneity in dyslexia. The participant pool consisted of seven compensated dyslexic adults and seven age and IQ matched controls. The dyslexic group were impaired, relative to the control group, on measures of literacy, phonological awareness, working memory and processing speed. Magnetoencephalographic recordings were conducted during processing of simple, non-speech, auditory stimuli. Results confirm that low-level auditory processing deficits are present in compensated dyslexic adults. The amplitude of N1m responses to tone pair stimuli were reduced in the dyslexic group. However, there was no evidence that manipulating either the silent interval or the frequency separation between the tones had a greater detrimental effect on dyslexic participants specifically. Abnormal MMNm responses were recorded in response to frequency deviant stimuli in the dyslexic group. In addition, complete stimulus omissions, which evoked MMNm responses in all control participants, failed to elicit significant MMNm responses in all but one of the dyslexic individuals. The data indicate both a deficit of frequency resolution at a local level of auditory processing and a higher-level deficit relating to the grouping of auditory stimuli, relevant for auditory scene analysis. Implications and directions for future research are outlined.

Simultaneous en-face imaging of two layers in human retina by low-coherence reflectometry

We show that with a fiberized multiple Michelson-interferometer-type configuration, transverse images from several layers in the human eye can be simultaneously obtained. We demonstrate the principle by producing simultaneous 100×100 pixel en-face images of a 4 mm×4 mm region on a postmortem retina for two depth positions 250 µm apart.

Low level feature detection in human vision

Influential models of edge detection have generally supposed that an edge is detected at peaks in the 1st derivative of the luminance profile, or at zero-crossings in the 2nd derivative. However, when presented with blurred triangle-wave images, observers consistently marked edges not at these locations, but at peaks in the 3rd derivative. This new phenomenon, termed ‘Mach edges’ persisted when a luminance ramp was added to the blurred triangle-wave. Modelling of these Mach edge detection data required the addition of a physiologically plausible filter, prior to the 3rd derivative computation. A viable alternative model was examined, on the basis of data obtained with short-duration, high spatial-frequency stimuli. Detection and feature-making methods were used to examine the perception of Mach bands in an image set that spanned a range of Mach band detectabilities. A scale-space model that computed edge and bar features in parallel provided a better fit to the data than 4 competing models that combined information across scale in a different manner, or computed edge or bar features at a single scale. The perception of luminance bars was examined in 2 experiments. Data for one image-set suggested a simple rule for perception of a small Gaussian bar on a larger inverted Gaussian bar background. In previous research, discriminability (d’) has typically been reported to be a power function of contrast, where the exponent (p) is 2 to 3. However, using bar, grating, and Gaussian edge stimuli, with several methodologies, values of p were obtained that ranged from 1 to 1.7 across 6 experiments. This novel finding was explained by appealing to low stimulus uncertainty, or a near-linear transducer.