Caterina Ripamonti

Long-term effect of gene therapy on Leber's congenital amaurosis.

BACKGROUND Mutations in RPE65 cause Lebers congenital amaurosis, a progressive retinal degenerative disease that severely impairs sight in children. Gene therapy can result in modest improvements in night vision, but knowledge of its efficacy in humans is limited. METHODS We performed a phase 1-2 open-label trial involving 12 participants to evaluate the safety and efficacy of gene therapy with a recombinant adeno-associated virus 2/2 (rAAV2/2) vector carrying the RPE65 complementary DNA, and measured visual function over the course of 3 years. Four participants were administered a lower dose of the vector, and 8 were administered a higher dose. In a parallel study in dogs, we investigated the relationship among vector dose, visual function, and electroretinography (ERG) findings. RESULTS Improvements in retinal sensitivity were evident, to varying extents, in six participants for up to 3 years, peaking at 6 to 12 months after treatment and then declining. No associated improvement in retinal function was detected by means of ERG. Three participants had intraocular inflammation, and two had clinically significant deterioration of visual acuity. The reduction in central retinal thickness varied among participants. In dogs, RPE65 gene therapy with the same vector at lower doses improved vision-guided behavior, but only higher doses resulted in improvements in retinal function that were detectable with the use of ERG. CONCLUSIONS Gene therapy with rAAV2/2 RPE65 vector improved retinal sensitivity, albeit modestly and temporarily. Comparison with the results obtained in the dog model indicates that there is a species difference in the amount of RPE65 required to drive the visual cycle and that the demand for RPE65 in affected persons was not met to the extent required for a durable, robust effect. (Funded by the National Institute for Health Research and others; ClinicalTrials.gov number, NCT00643747.).

Measurements of the effect of surface slant on perceived lightness

When a planar object is rotated with respect to a directional light source, the reflected luminance changes. If surface lightness is to be a reliable guide to surface identity, observers must compensate for such changes. To the extent they do, observers are said to be lightness constant. We report data from a lightness matching task that assesses lightness constancy with respect to changes in object slant. On each trial, observers viewed an achromatic standard object and indicated the best match from a palette of 36 grayscale samples. The standard object and the palette were visible simultaneously within an experimental chamber. The chamber illumination was provided from above by a theater stage lamp. The standard objects were uniformly-painted flat cards. Different groups of naive observers made matches under two sets of instructions. In the Neutral Instructions, observers were asked to match the appearance of the standard and palette sample. In the Paint Instructions, observers were asked to choose the palette sample that was painted the same as the standard. Several broad conclusions may be drawn from the results. First, data for most observers were neither luminance matches nor lightness constant matches. Second, there were large and reliable individual differences. To characterize these, a constancy index was obtained for each observer by comparing how well the data were accounted for by both luminance matching and lightness constancy. The index could take on values between 0 (luminance matching) and 1 (lightness constancy). Individual observer indices ranged between 0.17 and 0.63 with mean 0.40 and median 0.40. An auxiliary slant-matching experiment rules out variation in perceived slant as the source of the individual variability. Third, the effect of instructions was small compared to the inter-observer variability. Implications of the data for models of lightness perception are discussed.

An Equivalent Illuminant Model for the Effect of Surface Slant on Perceived Lightness

In the companion study (C. Ripamonti et al., 2004), we present data that measure the effect of surface slant on perceived lightness. Observers are neither perfectly lightness constant nor luminance matchers, and there is considerable individual variation in performance. This work develops a parametric model that accounts for how each observers lightness matches vary as a function of surface slant. The model is derived from consideration of an inverse optics calculation that could achieve constancy. The inverse optics calculation begins with parameters that describe the illumination geometry. If these parameters match those of the physical scene, the calculation achieves constancy. Deviations in the models parameters from those of the scene predict deviations from constancy. We used numerical search to fit the model to each observers data. The model accounts for the diverse range of results seen in the experimental data in a unified manner, and examination of its parameters allows interpretation of the data that goes beyond what is possible with the raw data alone.

Invariant cone-excitation ratios may predict transparency

Cone-excitation ratios for pairs of surfaces are almost invariant under changes in illumination and offer a possible basis for color constancy [Proc. R. Soc. London Ser. B 257, 115 (1994)]. We extend this idea to the perception of transparency on the basis of the close analogy between the changes in color signals that occur for surfaces when the illumination changes and the changes in color signals when the surfaces are covered by a filter. This study presents measurements and simulations to investigate the conditions under which cone-excitation ratios are statistically invariant for physically transparent systems. The invariance breaks down when the spectral transmission of the filters is low at some or all wavelengths. We suggest that cone-excitation ratios might be useful to define the stimulus conditions necessary for the perception of transparency.

The effect of sildenafil citrate (Viagra®) on visual sensitivity

The erectile dysfunction medicine sildenafil citrate (Viagra) inhibits phosphodiesterase type 6 (PDE6), an essential enzyme involved in the activation and modulation of the phototransduction cascade. Although Viagra might thus be expected to impair visual performance, reports of deficits following its ingestion have so far been largely inconclusive or anecdotal. Here, we adopt tests sensitive to the slowing of the visual response likely to result from the inhibition of PDE6. We measured temporal acuity (critical fusion frequency) and modulation sensitivity in four subjects before and after the ingestion of a 100-mg dose of Viagra under conditions chosen to isolate the responses of either their short-wavelength-sensitive (S-) cone photoreceptors or their long- and middle-wavelength-sensitive (L- and M-) cones. When vision was mediated by S-cones, all subjects exhibited some statistically significant losses in sensitivity, which varied from mild to moderate. The two individuals who showed the largest S-cone sensitivity losses also showed comparable losses when their vision was mediated by the L- and M-cones. Some of the losses appear to increase with frequency, which is broadly consistent with Viagra interfering with the ability of PDE6 to shorten the time over which the visual system integrates signals as the light level increases. However, others appear to represent a roughly frequency-independent attenuation of the visual signal, which might also be consistent with Viagra lengthening the integration time (because it has the effect of increasing the effectiveness of steady background lights), but such changes are also open to other interpretations. Even for the more affected observers, however, Viagra is unlikely to impair common visual tasks, except under conditions of reduced visibility when objects are already near visual threshold.

Classical and inverted White's effects.

In classical Whites effect, intermediate-luminance targets appear lighter when they interrupt the dark stripes of a grating and darker when they interrupt the light stripes. The effect is reversed when targets are of double-increment or double-decrement luminance, relative to the luminances of grating stripes. To find a common explanation for classical and inverted effects, we ran two experiments. In experiment 1, we utilised intermediate-target displays to show that perceived transparency dominates over occlusion only when the target luminance is close to the luminances of top regions. This result weakens transparency-based accounts of Whites effect. In experiment 2, we varied grating contrast and target luminance to measure the classical effect in seven intermediate-target cases, as well as the inverted effect in four double-increment and four double-decrement cases. Both types of effect are explained by a common model, based on assimilation to the top region and contrast with the interrupted region, weighted by adjacency along the luminance continuum.

The loss of the PDE6 deactivating enzyme, RGS9, results in precocious light adaptation at low light levels

The GTPase activating protein, RGS9-1, is vital for the deactivation and regulation of the phototransduction cascade (C. K. Chen et al., 2000; C. W. Cowan, R. N. Fariss, I. Sokal, K. Palczewski, & T. G. Wensel, 1998; W. He, C. W. Cowan, & T. G. Wensel, 1998; A. L. Lyubarsky et al., 2001). Its loss through genetic defects in humans has been linked to a slow recovery to changes in illumination (K. M. Nishiguchi et al., 2004). Such a deficit is to be expected because RGS9-1 normally speeds up the deactivation of the activated phosphodiesterase effector molecule, PDE6*, and thus accelerates the turning off of the visual response. Paradoxically, however, we find that the cone response in an observer lacking RGS9-1 is faster at lower light levels than it is in a normal observer. Though surprising, this result is nonetheless consistent with molecular models of light adaptation (e.g., E. N. Pugh, S. Nikonov, & T. D. Lamb, 1999), which predict that the excess of PDE6* resulting from the loss of RGS9-1 will shorten the visual integration time and speed up the visual response at inappropriately low light levels. The gain in speed caused by the superfluity of PDE6* at lower light levels compensates for the loss caused by its slow deactivation; thus quickening the response relative to that in the normal. As the light level is increased and the PDE6* concentration in the normal rises relative to that in the observer lacking RGS9-1, the temporal advantage of the latter is soon lost, leaving only the deficit due to delayed deactivation.

Evaluation of a multiscale color model for visual difference prediction

How different are two images when viewed by a human observer? There is a class of computational models which attempt to predict perceived differences between subtly different images. These are derived from theoretical considerations of human vision and are mostly validated from psychophysical experiments on stimuli, such as sinusoidal gratings. We are developing a model of visual difference prediction, based on multiscale analysis of local contrast, to be tested with psychophysical discrimination experiments on natural-scene stimuli. Here, we extend our model to account for differences in the chromatic domain by modeling differences in the luminance domain and in two opponent chromatic domains. We describe psychophysical measurements of objective (discrimination thresholds) and subjective (magnitude estimations) perceptual differences between visual stimuli derived from colored photographs of natural scenes. We use one set of psychophysical data to determine the best parameters for the model and then determine the extent to which the model generalizes to other experimental data. In particular, we show that the cues from different spatial scales and from the separate luminance and chromatic channels contribute roughly equally to discrimination and that these several cues are combined in a relatively straightforward manner. In general, the model provides good predictions of both threshold and suprathreshold image differences arising from a wide variety of geometrical and optical manipulations. This implies that models of this class can be generally useful in specifying how different two similar images will look to human observers.

Conditions for perceptual transparency

We review the conditions that are necessary for the perception of transparency and describe the spatiochromatic constraints for achromatic and chromatic transparent displays. These constraints can be represented by the convergence model and are supported by psychophysical data. We present an alternative representation of the constraints necessary for transparency perception that is based on an analogy with a model of colour constancy and the invariance of cone-excitation ratios. Recent psychophysical experiments are described that suggest that displays where the cone-excitation ratios are invariant produce a stronger impression of transparency than displays where the cone excitations are convergent. We argue that the spatial relations in an image are preserved when a Mondrian-like surface is partially covered by a transparent filter and therefore show an intriguing link between transparency perception and colour constancy. Finally, we describe experiments to relate the strength of the transparency percept with the number of unique patches in the image display. We find that the greater the number of surfaces in the display that are partially covered by a transparent filter the stronger the impression of transparency.

A multiresolution color model for visual difference prediction

How different are two images when viewed by a human observer? Such knowledge is needed in many situations including when one has to judge the degree to which a graphics representation may be similar to a high-quality photograph of the original scene. There is a class of computational models which attempt to predict such perceived differences. These are derived from theoretical considerations of human vision and are mostly validated from experiments on stimuli such as sinusoidal gratings. We are developing a model of visual difference prediction based on multi-scale analysis of local contrast, to be tested with psychophysical discrimination experiments on natural-scene stimuli. Here, we extend our model to account for differences in the chromatic domain. We describe the model, how it has been derived and how we attempt to validate it psychophysically for monochrome and chromatic images.