Dose-response of the cultured bovine lens to butyl, methyl and propyl parabens.

Pre-screening of cosmetic ingredients is vital for consumer safety. Previous in vivo techniques, such as the Draize test, have proved to be unreliable in predicting ocular irritancy and therefore there is a need for alternate testing methodologies. One such test is the scanning laser in vitro assay system which quantifies irritancy based on the focusing ability of the cultured bovine lens. In combination with confocal microscopy, a more thorough documentation of ocular irritancy can be achieved. This study investigates the response of cultured bovine lenses over time to butyl, methyl and propyl parabens, which are common antimicrobial agents found in cosmetic and ophthalmic products. The focusing ability of the lens was measured with an automated laser scanner over a period of 96 h. At 120 h post-treatment, the lenses were analysed by using a confocal laser scanning microscope to determine the characteristics of nuclei, and the morphology and distribution of mitochondria within the lenses. Irritancy to the three parabens was investigated at both an optical and cellular level. Each of the parabens was tested at 0.002% and 0.2%, where the 0.2% butyl paraben was found to be the most irritating.

Long-term lens organ culture system with a method for monitoring lens optical quality.

The Scan Tox System is a method for monitoring lens optical quality (focus or lack of focus) in culture conditions, which mimic conditions inside the eye. The ocular lens is an ideal organ for long-term culture experiments because it has no direct blood supply and no connection to the nervous system. The Scan Tox System makes it possible to keep lenses for long-term studies of up to a few weeks. The use of cultured lenses, mainly bovine, replaces the need for testing the effects of potentially damaging agents on live animals. This optical monitoring apparatus uses a computer-operated scanning laser beam, a video-camera system and a video frame analyzer to record the focal length and transmittance of the cultured lens. The scanner is designed to measure the focal length at points across the diameter of the lens. The lens container permits the lens to be exposed to a vertical laser beam from below. The laser source projects its light onto a plain mirror, which is mounted at 45 degrees C on a carriage assembly. The mirror reflects the laser beam directly up through the test lens. The mirror carriage is connected to a positioning motor, which moves the laser beam across the lens. The camera sees the cross section of the beams and, by examining the image at each position of the mirror, Scan Tox software is able to measure the quality of the lens by calculating the back vertex distance for each beam position. The cultured lenses continue to maintain their original refractive function. When foreign substances are introduced to a cultured lens, the Scan Tox System measures the resulting optical response. This provides a very sensitive means to follow early damage to the eye lens. Because the lens is maintained in an intact state in solutions that are similar to those inside the eye, the lens retains its normal recuperative powers. So in addition to measuring early damage, this system allows measurement of recovery from damage.

Contact lens design for lateralizing visual input.

A contact lens designed for use in lateralizing visual input is described. The design includes a carrier soft contact lens into which a smaller hard contact lens is inserted. The hard lens is partially painted black to obstruct a portion of the visual field, as required for experimental purposes.

Aging and the optical quality of the rat crystalline lens.

Split laser beams of varying separation were directed through the crystalline lenses of rats of young, old, and intermediate age. The variation in back vertex distance was determined from photographs of the focal effects of each lens. The results indicate that negative spherical aberration of the rat lens increases with age. In addition, local reversals in the slopes of the negative spherical aberration curves are larger and more frequent in the older lenses. The increase in negative spherical aberration is likely due to tighter packing of central lens fibers and a consequent increase in central refractive index although change in lens shape may contribute as well.

Accommodation and chromatic aberration in young children.

Retinoscopy through colored filters (chromoretinoscopy) was used to determine the portion of the chromatic aberration interval in focus when young children (2 to 6 years of age) fixate at far and near. The results indicate that the children may be divided into three distinct groups. In the youngest group there is haphazard focusing within the chromatic aberration interval at far and near. The middle group shows selective focusing of the red end of the chromatic interval at both far and near. Children in the oldest group focus the red end when fixating at far and the green end when fixating at near, thereby sparing accommodation. These results suggest that the eye's use of the chromatic aberration interval to spare accommodation, as found in previous studies carried out on adults, is learned by about the fourth year of a child's life.

Lens optical quality is a direct function of lens sutural architecture.

We analyzed the structural and functional relationship between lens sutures and lens optical quality (focal length variability) by correlative scanning electron microscopy (SEM) and laser scan analysis. Twenty-two rabbit lenses (8 pigmented and 14 albino) were used in this study. Lenses were initially scanned by a low-power helium-neon laser beam that was passed either at an acute angle to a lens suture or along a lens suture. The results of laser scan analysis with the incident beam passed at an acute angle to a lens suture showed that generally, rabbit lenses were well corrected for spherical aberration. Subsequent SEM analysis showed that areas of lenses scanned that produced the least amount of focal variability were characterized by uniform fiber cells arranged in parallel, radial cell columns. In contrast, the results of laser scan analysis with the incident beam passed along a lens suture showed that there was significant focal length variability, i.e., spherical aberration at the lens sutures. Subsequent SEM analysis showed that the areas of lenses scanned that produced the greatest amounts of focal variability (lens sutures) were characterized by nonuniform fiber cell ends arranged as erratic suture branches in single growth shells and collectively as erratic suture planes formed between growth shells extending from the embryonic nucleus to the lens periphery. Furthermore, the amount of focal variability was directly proportional to the degree of structural disorder at the lens sutures. This is the first study to unequivocally show that the relationship between lens optical quality and specific parameters of lens morphology (lens sutures) can be quantified. These findings may help to elucidate the pathologic changes that lead to presbyopia and cortical cataractogenesis because these lenses are characterized by asymmetrical suture patterns and planes.

Effects of experimentally induced ametropia on the morphology and optical quality of the avian crystalline lens.

PURPOSE: To examine the effects of refractive error on avian lens morphology and optical quality. METHODS: Hatchling white leghorn chicks were unilaterally goggled for 7 days with either a form-deprivation goggle (n = 12), a -10 D defocus goggle (n = 12), or a +10 D defocus goggle (n = 12) to induce myopia and hyperopia. Optical quality of lenses (focal length and focal length variability) from treated and contralateral control eyes was assessed using a scanning laser apparatus. Lens morphology was examined by light and electron microscopy. RESULTS: Although the induction of refractive errors did not significantly alter lens size, shape, paraxial focal length, or average focal length, average focal length variability increased. Lenses from eyes goggled with form-deprivation and +10 D defocus goggles demonstrated a twofold increase in average focal length variability, when compared with their contralateral controls. The morphology of the lens is not altered by these experimental manipulations. CONCLUSIONS: This study provides evidence that the refractive development of the chick lens is not independent of the refractive development of the ocular globe and that chick lenticular development is influenced by both genetics and visual experience.

Effect of hydrogen peroxide, DL-propranolol, and prednisone on bovine lens optical function in culture.

Lens refractive function was monitored during long-term bovine lens culture experiments in which low concentrations of potentially damaging agents were added to the culture media. The agents tested were the drugs DL-propranolol and prednisone, and hydrogen peroxide. A computer-driven scanning laser system was used to monitor lens focal length during culture. The system consists of a scanning helium-neon laser beam, a television video camera, and a video frame digitizer. The system first locates the optical center of the lens, defined as the position of little or no refractive deviation of the beam. The laser scans the lens in small steps (0.05 mm), while the digitizer measures focal length for each beam position. A graphic profile of lens focal variation is plotted. A concentration of 0.1 mM hydrogen peroxide produces no significant change in lens focus after 315 hr of incubation. When the concentration is increased 10-fold (1.0 mM), lens focal ability is radically disturbed after 60 hr of incubation. Lenses incubated with prednisone (0.03 mM) for up to 525 hr show no difference in focal characteristics in comparison to control lenses. Propranolol in a concentration of 0.1 mM disrupts lens focal ability after 250 hr of incubation, whereas a higher concentration (1.0 mM) produces a similar effect after only 150 hr. These results indicate that lens refractive function is a sensitive measure of lens function during culture.

Optical constancy of the chick lens during pre- and post-hatching ocular development.

Embryonic and post-embryonic development of the ocular lens is associated with the continual production of new secondary lens fibers by the mitotic activity of equatorial epithelial cells. This continual development affects lens size and shape and refractive index distribution. Study of embryonic lens optical function has been largely ignored. The optical characteristics of the developing chick lens, including paraxial and eccentric focal lengths, were measured during the embryonic period of development and up to 15 days after hatching. Measurements were made with an automated scanning laser system in which the video image of a helium-neon laser beam refracted by an excised lens in solution is digitized. Focal length is measured for beams moving in small steps on either side of that center. Measurements were made on excised lenses as well as with the lens in situ within the anterior segment of the eye. The results, collected from a study of a total of 80 lenses, indicates that embryo lenses at 6-7 days of incubation have long and very variable focal lengths. At the tenth embryo day, focal length drops by more than one-half and focal variations, between lenses and for different beam positions within a single lens, is reduced. Further measures for 14- and 17-day embryo lenses, as well as for lenses from hatchling and 5-, 10- and 15-day-old chicks, indicate that there is little change in focal length, either paraxially or for eccentric beam positions.(ABSTRACT TRUNCATED AT 250 WORDS)

Proteins of the vitreous humor during experimentally induced myopia.

Significant myopia was evident within 14 days after a translucent goggle was placed over the eye of a newly hatched chick. This myopia was characterized by large negative refractive error, increased axial and equatorial eye lengths, and increased wet eye weight. The volume of the vitreous humor increased in the myopic eye, because of a signficant accumulation of liquid vitreous humor. The protein concentration of the vitreous humor of the myopic and nonmyopic eyes was greater at day 14 than at day 0 (ie, first day after hatching). Moreover, over this time, the protein concentration in the liquid component decreased, whereas the protein concentration in the gel component increased in both myopic and nonmyopic eyes. No difference was found in the protein concentrations between the myopic and nonmyopic eye at day 14. Polyacrylamide gel electrophoresis revealed changes in band position and intensity of individual protein compoents between days 0 and 14. No major differences were found in the gel or the liquid vitreous humor between the myopic and the nonmyopic eyes. Comparison of vitreous humor proteins with proteins from the plasma showed that some but not all of the proteins in the vitreous had the same apparent molecular masses as proteins from these sources.

Action spectrum and recovery for in vitro UV-induced cataract using whole lenses.

PURPOSE: To establish the in vitro action spectrum for acute UV cataractogenesis using whole cultured lenses. The recovery pattern of the induced cataract was also investigated. METHODS: Aseptically dissected porcine lenses were cultured in glass chambers. At 1 week, lenses were exposed to a predetermined UV energy (J/cm(2)) at specific wavebands ranging from 270 to 370 nm at 5- and 10-nm intervals. The UV energy was generated by a PRA integrated arc lamp system using a water-cooled 1000 W, high-pressure xenon lamp. The lamp output was limited using a deionized water filter, a monochromator, and secondary optics. An electronic shutter was used to control the exposure time. The median effective dose, ED(50) (i.e., UV energy threshold) for each waveband was statistically determined using probit analysis. Irradiated spots (3.06 mm(2)) on the lenses were monitored every 6 to 12 hours up to 48 hours postirradiation for any UV-induced opacity with a dissecting microscope and photomicrography. The ED(50)s were plotted against wavelengths to obtain the action spectrum. RESULTS: The threshold values for 270, 300, and 365 nm were 0.057, 0.069, and 137.19 J/cm(2), respectively. Permanent UV-induced cataract was obtained at twice the threshold values for UVB and UVA. CONCLUSIONS: An action spectrum for in vitro UV-induced cataract using whole cultured lens is established. These data are comparable to published in vitro (with isolated lens epithelial cells) and in vivo action spectra. The recovery pattern appears to be similar to the in vivo situation.

The internalization of posterior subcapsular cataracts (PSCs) in Royal College of Surgeons (RCS) rats. II. The inter-relationship of optical quality and structure as a function of age.

PURPOSE: The Royal College of Surgeons (RCS) rat is an animal model for human retinal degenerative disease and posterior subcapsular cataracts (PSCs). The purpose of this study was to correlate the structure and optical quality of RCS lenses with PSCs as a function of their internalization, with normal, non-cataractous, age-matched control lenses. METHODS: Correlative light (LM), scanning electron microscopic (SEM), three-dimensional computer assisted drawings (3D-CADs) and low power helium-neon laser scan analysis were used to examine the structure and function of lenses. RESULTS: The optical properties (average focal length variability; sharpness of focus) of RCS rat lenses are quantitatively compromised by PSCs. Correlative LM and SEM analysis of RCS lenses at various stages of PSC internalization (1.5, 3, 6, 9, 12 and 15 months of age), revealed that the sutures formed by additional fiber growth were progressively more abnormal. During PSC internalization, two to nine small suture branches were formed and arranged in modified line to multiple y configurations rather than the normal three branch y sutures. These temporal changes were also chronicled in animated 3D-CAD videos derived from lens reconstructions based on LM and SEM micrographs from the selected time points stated above. However, laser scan analysis also revealed that as the PSCs of RCS rat lenses were progressively internalized, there was a steady improvement in total sharpness of focus that reached normal levels by 12 months of age. The correlation of laser scan and structural data from specific regions of lenses revealed the following: 1. The abnormal posterior sutures of RCS rats with internalized PSCs effect a greater reduction in optical quality than normal posterior sutures of age-matched controls; 2. However, the resulting abnormal suture plane area was cumulatively similar to that of age-matched controls; 3. Thus, total optical quality was similar between RCS lenses with internalized PSCs and age-matched controls by 12 months of age. CONCLUSIONS: The results of this study show that RCS lenses with internalized PSCs can appear grossly, and indeed optically perform, at levels comparable to aged lenses. These findings are consistent with clinical observations of spontaneous recovery from PSC. The results suggest that human PSCs that occur as a consequence of retinal degenerative disease could also be the result of abnormal posterior suture growth. If this is proven to be the case, such PSCs may have some capacity for repair or recovery thereby obviating their surgical removal.

Optical function and mitochondrial metabolic properties in damage and recovery of bovine lens after in vitro carbonyl cyanide m-chlorophenylhydrazone treatment.

In order to elucidate the correlation between lens optical function and metabolic function, in vitro bovine lens optical quality and mitochondrial integrity was measured following treatment with carbonyl cyanide m-chlorophenylhydrazone (the mitochondrial depolarizing agent, CCCP). The results indicate that in vitro exposure to CCCP resulted in concentration and time-dependent loss of sharp focus. The concentrations tested included 65.0, 32.5, 16.25 and 8.125 microm CCCP. Lenses treated with two lower concentrations show recovery from damage at the 24-h scan point. In lenses treated with 65 microM CCCP, mitochondria in lens epithelial and superficial cortical fibre cells appeared short and swollen. The results of this study indicate that lens optical function and mitochondrial integrity are closely correlated.

Impairment of eye lens cell physiology and optics by broadband ultraviolet A-ultraviolet B radiation.

The phototoxicity of ultraviolet A (UVA) alone and UVA plus ultraviolet B (UVB) combined on cultured porcine lenses was investigated by analyzing cellular function as measured with a fluorescence bioassay approach and optical integrity, in terms of sharpness of the lens focus as measured with a scanning laser system. The bioassay consisted of carboxyfluorescein diacetate-acetoxymethyl ester and alamarBlue fluorescent dyes. Aseptically dissected porcine lenses were maintained in modified medium 199 without phenol red supplemented with 1% penicillin-streptomycin and 4% porcine serum. At 1 week of preincubation, baseline measurements were obtained. Then the lenses were treated with single exposures of different UVA and UVB energy levels. The lenses treated with 86 J/cm2 UVA alone showed a significant (P < 0.05) decrease in cellular and optical integrity at 48 h after exposure, whereas those treated with 43 J/cm2 UVA alone did not show significant phototoxic effect. Lenses treated with 15.63 J/cm2 UVA plus 0.019 J/cm2 UVB combined showed significant adverse effects beginning from 48 h after exposure. Also, there was no recovery. These findings show that a high UVA dose alone and relatively low UVA in combination with low UVB radiant exposure can impair lens cellular and optical functions, respectively.

The naturally occurring accommodative response of the oscar, Astronotus ocellatus, to visual stimuli.

Observation of the naturally-occurring accommodative response of a highly visual teleost species provides the opportunity to study its stimulus-response accuracy. A video recording system, linked to a computer digitizing program, was used to measure accommodative changes in lens position in the oscar, Astronotus ocellatus, in response to known food stimuli. The largest lens deviations occurred along an axis c. 22 deg from the pupillary plane. Consistent underaccommodation was reported for the closest of targets, a finding reminiscent of accommodative lag reported in human studies.

Spherical aberration of the crystalline lens.

Split beams of varying separations from a helium-neon laser were directed through the crystalline lenses of a number of vertebrates. Photographs of the focal effects indicate the extent to which the refractive index variation of the lens and lens shape control spherical aberration. Of the fish studied, only rock bass lenses are relatively free of spherical aberration. Both goldfish and yellow perch exhibit substantial amounts of positive spherical aberration. Varying amounts of negative spherical aberration are present in frog, juvenile duck, dog and rat lenses. Positive and negative spherical aberration is found in human and cat lenses while cow, pig, lamb and rabbit lenses are almost free of aberration.

Longitudinal chromatic aberration of the vertebrate eye.

A recent study involving Abbe and Pulfrich refractometry analyses the dispersion of the human lens and the ocular media of a number of vertebrates. In general, the lens and, to a lesser extent, the cornea, are more dispersive than expected at wavelengths below 500 nm. The dispersion findings of this study were used in conjunction with reduced eye parameters of a number of vertebrates to calculate the longitudinal chromatic aberration of rock bass, frog, chicken, rat, cat, pig, cow, and human eyes. The calculated chromatic aberration of the human eye is greater than values reported earlier, because of the exaggerated dispersion of the lens at short wavelengths. While the values calculated for the additional species studies may be larger in some instances than expected, presumably due to lens dispersion as well, chromatic aberration is not large enough to account for the hyperopia found by retinoscopic study of small eyes.

Penguin vision in air and water.

Refractive states measured by retinoscopy and photorefraction indicate that rockhopper (Eudyptes crestatus), Magellanic (Spheniscus magellanicus) and gentoo (Pygoscelis papua) penguins are approximately emmetropic in air and water. Extensive myopia in air, as predicted by early authors, is nonexistent. Photorefractive measurements of refractive state in water indicate that rockhopper, gentoo, Magellanic and king (Aptenodytes patagonica) penguins can accommodate sufficiently to make up for the loss of refractive power of the cornea. Corneas of rockhopper and Megellanic penguins are flattened relative to the overall size of the eye. This feature minimizes the optical effect of submergence.

Chromatic dispersion of the ocular media.

Measurements of chromatic dispersion of aqueous and vitreous humors, cornea and lens of the eye are sparse and incomplete. The wavelength variation in refractive index of the ocular media of cow, pig, frog (Rana pipiens), chicken, rock bass (Ambloplites rupestris), albino rat and cat as well as human lenses was determined by means of Abbe and Pulfrich refractometry. While the humors are somewhat less dispersive than water, the cornea is more dispersive at short wavelengths. In general, the lens is significantly more dispersive than water with dispersion increasing asymptotically at the blue end of the spectrum. The exaggerated dispersion taking place at short wavelengths should be taken into account in calculations of chromatic aberration.

Optical development of the ocular lens of an elasmobranch, Raja eglanteria [corrected].

The shape and focal properties (spherical aberration) of the ocular lens of hatchling, yearling and adult clearnose skates (Raja eglanteria) were examined. In contrast to the spherical shape of the typical teleost lens, the lens of the clearnose skate is slightly aspherical; equatorial diameter being greater than axial diameter. The asphericity is approximately equal, in absolute measures, at all ages and is thus more obvious in younger skates. Lens spherical aberration is well corrected at all three levels of development. Yearling and adult lenses have relative focal lengths (Matthiessen's Ratio) which are close to the representative value of 2.55:1 expected for the teleost lens. However, the relative focal length of lenses from hatchling lenses is higher (about 2.75:1). This value approaches the upper limit of reports dealing with teleost lens focal characteristics. The difference between hatchling lenses and those of adult and yearling lenses may be due to the absence of extensive compression of central lens tissue by peripheral growth at early stages in development.