[The connective tissues, from the origin of the concept to its "Maturation" to extracellular matrix. Application to ocular tissues. Contribution to the history of medical sciences]. / Les tissus conjonctifs, de l'origine du concept à sa « mutation ¼ en matrice extracellulaire. Application aux tissus oculaires. Contribution à l'histoire des sciences médicales.

The "Tissue" concept emerged apparently in the medical literature at about the French revolution, during the second half of the 18(th) century. It was found in the texts written by the physicians of Béarn and Montpellier, the Bordeu-s and also by the famous physician, Felix Vicq d'Azyr, the last attending physician of the queen Marie-Antoinette, "Bordeu et al. (1775) et Pouliquen (2009)". It was elaborated into a coherent doctrine somewhat later by Xavier Bichat, considered as the founder of modern pathological anatomy, Bichat. With the advent of histochemistry, from the beginning of the 20(th) century, several of the principal macromolecular components of connective tissues, collagens, elastin, "acid mucopolysaccharides" (later glycosaminoglycans and proteoglycans) and finally structural glycoproteins were characterized. These constituents of connective tissues were then designated as components of the extracellular matrix (ECM), closely associated to the cellular components of these tissues by adhesive (structural) glycoproteins as fibronectin, several others and cell receptors, "recognising" ECM-components as integrins, the elastin-receptor and others. This molecular arrangement fastens cells to the ECM-components they synthesize and mediates the exchange of informations between the cells to the ECM (inside-out) and also from the ECM-components to the cells (outside-in). This macromolecular arrangement is specific for each tissue as a result of the differentiation of their cellular components. It is also the basis and condition of the fulfillment of the specific functions of differentiated tissues. This is a short description of the passage of the "tissue" concept from its vague origin towards its precise identification at the cellular and molecular level up to the recognition of its functional importance and its establishment as an autonomous science. This can be considered as a new example of the importance of metaphors for the progress of science, Keller (1995).

Classical LCAT deficiency resulting from a novel homozygous dinucleotide deletion in exon 4 of the human lecithin: cholesterol acyltransferase gene causing a frameshift and stop codon at residue 144.

Lecithin: cholesterolacyltransferase (LCAT) transacylates the fatty acid at the sn-2 position of lecithin to the 3beta-OH group of cholesterol forming lysolecithin and the majority of cholesteryl ester found in plasma. LCAT participates in the reverse cholesterol transport pathway in man where it esterifies tissue-derived cholesterol following efflux from peripheral cells into HDL. Only 38 unique mutations in the human LCAT gene have been reported worldwide. Our French female proband presented with corneal opacity and no detectable plasma LCAT activity using either endogenous or exogenous assays. Her total plasma cholesterol and HDL cholesterol were low (2.34 mmol/l and 0.184 mmol/l, respectively) with a very high cholesterol/cholesteryl ester molar ratio (10.9:1). Plasma triglycerides were 0.470 mmol/l with low apo B (40.5 mg/dl), apo A-I (14.7 mg/dl), apo A-II (6.8 mg/dl) and apo E (2.1 mg/dl) levels. Plasma lipoprotein analysis by ultracentrifugation showed very low HDL concentrations and a characteristic shift of the lipoprotein profile towards larger, less dense particles. No proteinuria, renal dysfunction or signs of atherosclerosis were noted at age 45. Sequence analysis of her LCAT gene showed a novel homozygous TG-deletion at residues 138-139 that resulted in a frameshift causing the generation of a stop codon and premature termination of the LCAT protein at amino acid residue 144. Western blotting of the patient's plasma using a polyclonal IgY primary antibody against human LCAT failed to demonstrate the presence of a truncated LCAT protein. A 53 bp mismatched PCR primer was designed to generate an Fsp 1 restriction site in the wild type sequence of exon 4 where the mutation occurred. The 155 bp PCR product from the wild type allele produced a 103 bp and 52 bp fragment with Fsp 1 and no cleavage products with the mutant allele thus permitting rapid screening for this novel mutation.

Retinal S-antigen in human subretinal fluid.

Studies in experimental models of retinal detachment have proposed that the degree of visual recovery following retinal reattachment depends upon the extent of photoreceptor degeneration. A means of assessing this degeneration would help in establishing postoperative prognosis. S-antigen (S-Ag) is a unique retinal protein found in outer segment disc membranes and photoreceptor cells. In 36 cases of human rhegmatogenous retinal detachment, subretinal fluid (SRF) concentrations of S-Ag, measured by radioimmunoassay, ranged from 43 to 170 ng/ml (serum: 1-28 ng/ml). Analysis of variance showed a positive correlation with the duration of detachment (P less than 0.001). There was a two-fold increase in S-Ag concentrations during the first 2 weeks of detachment (P less than 0.005), with constant levels thereafter. These findings reflect progressive photoreceptor degeneration and/or ongoing synthesis of outer segment proteins in the detached retina that stop after the second week of detachment. SRF S-Ag levels may provide a prognostic indicator of visual recovery after reattachment as well as a sensitive measure of retinal metabolic activity during detachment.

Dibutyryl cyclic adenosine monophosphate and forskolin alter the paracellular pathway in cultured corneal endothelial cells.

PURPOSE: This study describes the effects of the cyclic adenosine monophosphate (cAMP) pathway on the tight junctional barrier of the corneal endothelium, which plays a critical role in maintaining the corneal stroma in an underhydrated, transparent state. METHODS: Subcultured bovine corneal endothelial cells grown on filters were used to study the effects of dibutyryl-cAMP and forskolin on transendothelial electrical resistance and [3H]inulin flux. The tight junction-associated protein ZO-1 (zonula occludens protein-1) and F-actin were visualized by indirect immunofluorescence, and the ultrastructural organization of junctional complexes was studied by freeze-fracture electron microscopy. RESULTS: Cells formed a continuous monolayer of closely apposed hexagonal-type cells separated by a discontinuous belt of tight junctions with a transendothelial electrical resistance of 20.8 +/- 0.6 omega.cm2. Dibutyryl-cAMP (10(-4) M) and forskolin (10(-5) M) increased cell cAMP, significantly decreased the transendothelial resistance by 54% and 43%, respectively, and increased the flux of [3H]inulin from the apical to the basal side of the cells by 56% and 40%, respectively. Both agents also induced condensation of F-actin at the cell borders without any marked changes in the immunostaining of ZO-1 that delineated cell peripheries. However, freeze-fracture studies showed that dibutyryl-cAMP and forskolin induced dispersion of the tight junction network. CONCLUSIONS: These data suggest that activation of the cAMP-dependent pathway, leading to structural changes of the tight junctional network, may modulate the passive fluxes mediated by the paracellular pathway of the corneal endothelial barrier.

Reduction of corneal edema in endotoxin-induced uveitis after application of L-NAME as nitric oxide synthase inhibitor in rats by iontophoresis.

PURPOSE: To investigate the involvement of the cornea during endotoxin-induced uveitis (EIU) in the rat and the effect of Ngamma-nitro-L-arginine methyl ester (L-NAME) as nitric oxide synthase (NOS) inhibitor, administered by iontophoresis. METHODS: EIU was induced in Lewis rats that were killed at 8 and 16 hours after lipopolysaccharide (LPS) injection. The severity of uveitis was evaluated clinically at 16 hours, and nitrite levels were evaluated in the aqueous humor at 8 hours. Corneal thickness was measured, 16 hours after LPS injection, on histologic sections using an image analyzer. Transmission electron microscopy (TEM) was used for fine analysis of the cornea. Transcorneoscleral iontophoresis of L-NAME (100 mM) was performed either at LPS injection or at 1 and 2 hours after LPS injection. RESULTS: At 16 hours after LPS injection, mean corneal thickness was 153.7+/-5.58 microm in the group of rats injected with LPS (n=8) compared with 126.89+/-11.11 microm in the saline-injected rats (n=8) (P < 0.01). TEM showed stromal edema and signs of damage in the endothelial and epithelial layers. In the group of rats treated by three successive iontophoreses of L-NAME (n=8), corneal thickness was 125.24+/-10.36 microm compared with 146.76+/-7.52 microm in the group of rats treated with iontophoresis of saline (n=8), (P=0.015). TEM observation showed a reduction of stromal edema and a normal endothelium. Nitrite levels in the aqueous humor were significantly reduced at 8 hours by L-NAME treatment (P=0.03). No effect on corneal edema was observed after a single iontophoresis of L-NAME at LPS injection (P=0.19). Iontophoresis of saline by itself induced no change in corneal thickness nor in TEM structure analysis compared with normal rats. CONCLUSIONS: Corneal edema is observed during EIU. This edema is significantly reduced by three successive iontophoreses of L-NAME, which partially inhibited the inflammation. A role of nitric oxide in the corneal endothelium functions may explain the antiedematous effect of L-NAME.

Cytotoxic effects of FGF2-saporin on bovine epithelial lens cells in vitro.

PURPOSE: To test the ability of two preparations of FGF2-saporin, either FGF2 chemically conjugated to saporin (FGF2-SAP) or genetically engineered FGF2-saporin (rFGF2-SAP) to inhibit the growth of bovine epithelial lens (BEL) cells in vitro when in solution and when immobilized on heparin surface-modified (HSM) polymethylmethacrylate (PMMA) intraocular lenses (IOLs). METHOD: Bovine epithelial lens cells were incubated with various concentrations FGF2-saporin for as long as 4 days. The number of surviving cells was determined by counting the number of nuclei. Because FGF2 binds to heparin, FGF2-saporin was incubated with HSM PMMA IOLs; excess toxin was washed off, and the BEL cells were grown on the FGF2-saporin-treated IOLs (HSM and non-HSM) for 4 days. Cell density was determined by image analysis. RESULTS: Both FGF2-SAP and rFGF2-SAP were highly cytotoxic (nM range), with rFGF2-SAP 10 times less active than FGF2-SAP. FGF2-saporin bound to the surface of HSM IOLs and eluted by 2M NaCl retained its activity. Toxin bound to HSM IOLs killed more than 90% of the BEL cells placed on the IOL surface within 4 days. The ability of FGF2-saporin to prevent the growth of cells on the IOL surface was strictly dependent on the presence of heparin on the IOL. CONCLUSIONS: FGF2-saporin is bound to HSM PMMA IOLs and prevents the growth of epithelial cells on the surface of the lens.

In vivo inhibition of lens regrowth by fibroblast growth factor 2-saporin.

PURPOSE: To investigate the ability of fibroblast growth factor (FGF) 2-saporin to prevent lens regrowth in the rabbit. METHODS: Chemically conjugated and genetically fused FGF2-saporin (made in Escherichia coli) were used. Extracapsular extraction of the lens was performed on the rabbit, and the cytotoxin either was injected directly into the capsule bag or was administered by FGF2-saporin-coated, heparin surface-modified (HSM) polymethylmethacrylate intraocular lenses. The potential of the conjugate was checked by slit lamp evaluation of capsular opacification and by measuring crystallin synthesis. Toxin diffusion and sites of toxin binding were assessed by immunohistochemistry. Possible toxicity was determined by histologic analysis of ocular tissues. RESULTS: FGF2-saporin effectively inhibited lens regrowth when it was injected directly into the capsular bag. However, high concentration of the toxin induced transient corneal edema and loss of pigment in the iris. Intraocular lenses coated with FGF2-saporin reduced lens regrowth and crystallin synthesis without any detectable clinical side effect. After implantation, FGF2-saporin was shown to have bound to the capsules and, to a lesser extent, to the iris; no histologic damage was found on ocular tissues as a result of implantation of drug-loaded HSM intraocular lenses. CONCLUSIONS: Chemically conjugated (FGF2-SAP) and genetically fused FGF2-saporin (rFGF2-SAP) bound to HSM intraocular lenses can prevent lens regrowth in the rabbit.

Antibiotics and corneal epithelial wound healing.

The healing of a de-epithelialized cornea was measured by serial standardized photography after treatment with different antibiotics. Histological studies were also performed. Low doses of bacitracin (500 units/mL), gentamicin sulfate (3 mg/mL), neomycin (3.5 mg/mL), and chloramphenicol (4 mg/mL) did not retard normal epithelial healing. However, higher doses of bacitracin (10,000 units/mL), gentamicin sulfate (10 mg/mL), and neomycin (8 mg/mL) significantly inhibited reepithelialization.

Excimer laser keratectomy for myopia with a rotating-slit delivery system.

We performed argon fluoride excimer laser (193-nm) superficial keratectomy for myopia on human donor eyes and on a resected corneal disc. The laser beam was shaped by a rotating slit to produce a circular ablation 7.5 mm in diameter, with a mathematically defined profile to correct myopia. The fluence at the surface of the cornea was 200 mJ/cm2; the laser was fired at 20 Hz. Each 4.5-mJ laser pulse etched a 0.17-micron deep image of the slit in the cornea. Since the slit moved (0.03 Hz), each successive pulse etched an area adjacent to the previous one, reducing damage from repetitive pulses striking the same area. The slit scanned the cornea many times and the summation of these individual ablations produced the smooth myopic ablation profile, as shown by computerized keratographs and light and electron microscopy.

Corneal wound healing in monkeys after repeated excimer laser photorefractive keratectomy.

Five rhesus monkey eyes underwent repeated argon fluoride (193 nm) excimer laser myopic photorefractive keratectomy 3 months following an initial ablation that had produced mild subepithelial haze. At 3 months all eyes had development of a dense subepithelial opacity and a thickened epithelium (12 cells, 80 microns) with vacuolization of basal cells, fragmented basement membrane, and a layer of subepithelial fibrosis containing activated fibroblasts. By 6 months the opacity was clearing; epithelium was thinner (50 microns); subepithelial fibrosis was more lamellar. By 15 months only mild haze persisted clinically; epithelium was 30 microns thick, with persistent basal vacuolization and focal basement membrane disruption; subepithelial fibrous tissue was more organized. Early repeated excimer laser ablation of the monkey cornea apparently induces vigorous stromal wound healing. Use of shallower ablations, corticosteroids, or a longer delay between ablations may be necessary for repeated laser surgery to be practical clinically.

Keratoprosthesis with biocolonizable microporous fluorocarbon haptic. Preliminary results in a 24-patient study.

BACKGROUND: Most complications of a keratoprosthesis occur at the tissue-to-implant interface. The ideal prosthesis would eliminate this interface by having the tissue actually grow into the supporting material. We present a prospective clinical human study of a novel biocolonizable keratoprosthesis in 24 eyes of 24 patients. DESIGN: To promote implant stability, the 9-mm-diameter haptic was fashioned using a custom-made microporous fluorocarbon with a 4-mm-diameter, 2.67-mm-long, central optic made of medical grade polymethylmethacrylate, giving a global visual field of 110 degrees to 130 degrees. Only bilaterally blind patients with untreatable corneal diseases were included in the study. The haptic was inserted into a lamellar pocket delaminated in the stroma, and the optic was positioned through a hole trephined in the central cornea. RESULTS: The average follow-up was 15.7 months (range, 4 to 28 months). The host corneal fibroblasts penetrated and proliferated into the peripheral microporous fluorocarbon and provided anchorage between the cornea and prosthesis. Seventeen patients (70.8%) had visual acuity improvements. Mean corrected final visual acuity was 20/100 (range, 20/30 to 20/400). Five anatomic failures occurred in the first 6 months (three extrusions, one dislocation of the optic, and one endophthalmitis). We had one case (4.1%) of treatable glaucoma. We successfully removed four of five retroprosthetic membranes that had occurred. No retinal detachment occurred. CONCLUSION: The biocompatible inert microporous polymer did not eliminate all mechanical complications associated with a keratoprosthesis.

Wound healing after excimer laser keratomileusis (photorefractive keratectomy) in monkeys.

Laser myopic keratomileusis (photorefractive keratectomy) was performed on 29 rhesus monkey corneas with an argon fluoride (193-nm) excimer laser and a computer-controlled, moving slit delivery system. The 4-mm-diameter central ablation zone ranged in depth from 11 microns (-2 diopters effect) to 46 microns (-8 diopters effect). Corneas were studied for the 9 months postoperatively by clinical slit-lamp microscopy, and periodically with light and transmission electron microscopy. By 6 weeks, mild to moderate subepithelial haze was apparent in 93% of the corneas, with considerable variability in density. Progressive clearing occurred so that by 6 to 9 months 12 of 13 surviving corneas (92%) were either completely clear (4 corneas) or trace hazy (8 corneas). The epithelium was thickened at 21 days after ablation and returned to normal thickness by 3 months. At 3 weeks, subepithelial fibroblasts were three times the density of normal keratocytes and returned to nearly normal numbers by 9 months. We concluded that the anterior monkey cornea demonstrated a mild, typical wound healing response after excimer laser keratomileusis.

Corneal stromal wound healing in rabbits after 193-nm excimer laser surface ablation.

An argon fluoride excimer laser (193 nm) with a moving slit delivery system was used to perform anterior myopic keratomileusis in both eyes of 24 New Zealand white rabbits. Rabbits were killed immediately after ablation and at intervals up to 100 days. By slit-lamp microscopy, four rabbits at day 100 exhibited four clear corneas and four corneas had central, spotty, subepithelial haze. Light and electron microscopy documented corneal healing. In the early stages a transient acellular zone in the anterior stroma appeared over a period of three weeks, followed by an increased number of fibrocytes. In the corneas with opacification, focal areas of 20-microns-thick subepithelial scarring were present. An unexpected finding was transient damage to posterior stromal keratocytes and endothelial cells. The endothelium produced a layer of granular material that migrated anteriorly across Descemet's membrane. Immunochemistry at day 6 showed a marked staining for collagen IV, proteoglycans, fibronectin, and laminin.

PMMA model of steep central islands induced by excimer laser photorefractive keratectomy.

We designed a polymethyl methacrylate (PMMA) model of refractive steep central islands (CIs) induced by PRK. A standardized photorefractive ablation procedure was performed using seven different excimer lasers on test PMMA specimens including 70 flat plates and 98 convex contact lenses. The resulting surface was analyzed by high-resolution confocal microscopy and computerized videokeratoscopy using both TMS-1 and CAS-2000 systems. A total of 50 (54.9%) CIs were observed using computerized videokeratoscopy. The rate of occurrence of CIs was significantly reduced by shock wave absorption (P = 0.0001), aspiration of fumes (P = 0.0044), and smaller diameter ablation (P = 0.0296). The diameter of the CIIs was significantly increased for broad-beam mode ablation (P = 0.016) and for larger ablation zones, (P = 0.042). The refractive power of CIs was significantly increased in the absence of a shock wave absorption system (P = 0.001). Only 20 (40%) of the CIs detected by the TMS-1 device were identified on CAS-2000 at a 0.5 diopter (D) scale resolution level. Shock wave induced deformation and subsequent dynamic alteration of convection forces applied to emitted particles may be the primary mechanism underlying the formation of CIs after PRK, regardless of any biological response of the ablated tissue. Reported rates and characteristics of CIs may largely depend upon the specific design of videokeratoscopes.

Parameters influencing human mu opioid receptor over-expression in baculovirus-infected insect cells.

The cDNA encoding the human mu opioid receptor (hMOR) was cloned in the baculovirus Autographa californica (AcMNPV) under the control of the polyhedrin promoter. We investigated the influence of different molecular constructions on receptor expression levels: the receptor was fused either to an amino- or a carboxy-terminal histidine tag (hMOR-N-His and hMOR-C-His respectively), or to the cleavable sequence signal of the baculovirus gp64 glycoprotein (gp-hMOR and gp-hMOR-C-His). Two cell lines, Spodoptera frugiperda (Sf9) and Trichoplusia ni (BTI-TN-5B1-4), in combination with three different culture media were also tested for their ability to produce maximal protein expression. Molecular constructions and culture conditions were both shown to influence substantially protein production. The best results were obtained using cells adapted to serum-free medium combined with constructions in fusion with the endogenous signal sequence of the baculovirus gp64 protein. Those conditions led to maximal expression and shortened the time required for receptor production. We also showed that an amino-terminal location of a hexahistidine tag was more detrimental to the expression level than a carboxy-terminal position.

Videokeratoscopy of recipient peripheral corneas in combined penetrating keratoplasty, cataract extraction, and lens implantation.

PURPOSE: We performed a prospective clinical trial to evaluate computerized videokeratoscopic analysis of the peripheral recipient cornea in intraocular lens power calculations for triple procedures: penetrating keratoplasty, cataract extraction, and intraocular lens insertion. METHODS: Patients with Fuchs' dystrophy underwent consecutive triple procedures. Surgery was performed in 16 eyes by a single surgeon (O.N.S.) using a single technique. If videokeratoscopic analysis disclosed dioptric powers greater than 40 diopters in the circumference of the corneal map, the surgeon's average postoperative central corneal power of 46 diopters was used with the regression formula. If dioptric powers less than 40 diopters were detected in the circumference of the corneal map, 45 diopters was used to avoid postoperative hyperopic shifts and to decrease deviation from intended refractive error. Refraction and videokeratoscopic analysis were performed six months after suture removal (18 to 24 months postoperatively). RESULTS: Analysis of covariance demonstrated that preoperative peripheral videokeratoscopic data of the recipient cornea correlated (P = .0001) with postoperative central corneal power, whereas preoperative central corneal power of the recipient cornea did not correlate (P = .35). Deviation from intended refraction (range, -2.54 to +1.22 diopters) was within 2 diopters in 14 eyes (88%) and within 3 diopters in all eyes. No patients had anisometropia greater than 3 diopters. CONCLUSION: Preoperative data from computerized videokeratoscopic analysis of the recipient peripheral cornea correlated with postoperative central corneal power, and improved postoperative refractive outcomes compared with previously reported results of triple procedures.

Ultrasound biomicroscopy of Baikoff anterior chamber phakic intraocular lenses.

BACKGROUND: The purpose of the study was to analyze qualitatively and quantitatively anatomic relationships of anterior chamber Baikoff phakic intraocular lenses (IOL) to the cornea, angle, iris, and lens in myopic eyes. METHODS: Thirteen phakic myopic eyes (-9.00 to -15.00 diopters) corrected by minus power, angle-supported, anterior chamber intraocular lenses of the ZB5M style (Chiron-Domilens), with a mean follow-up of 25 months (range 18 to 36 months), were examined clinically and by ultrasound biomicroscopy. RESULTS: The mean distance (+/- SD) between the central cornea and the IOL was 2.05 +/- 0.18 mm (range 1.8 to 2.47 mm); mean distance between the corneal periphery and the IOL was 1.56 +/- 0.17 mm (range 1.33 to 1.95 mm); and the mean distance between the IOL and the lens was 0.58 +/- 0.12 mm (range 0.48 to 0.81 mm). IOL footplates seemed to be correctly positioned in the angle in all eyes. We observed no goniosynechiae. In six eyes, we found localized posterior indentation of the iris caused by the haptics. Four of these six eyes exhibited oval pupils associated with an oversized IOL. CONCLUSIONS: High frequency ultrasound biomicroscopy can define anatomic relationships of anterior chamber phakic IOLs, and help analyze the mechanisms of corneal and iris complications.

Penetrating keratoplasty for keratoconus: role of videokeratoscopy and trephine sizing.

PURPOSE: To evaluate whether determining graft-host trephine disparity on the basis of videokeratoscopic data of keratoconus patients having penetrating keratoplasty (PKP) reduces ametropia and to correlate preoperative videokeratoscopic values, posterior axial length (PAL), and trephine disparity with postoperative refractive outcomes. SETTING: Hôtel-Dieu, University of Paris, France. METHODS: This randomized clinical trial comprised 18 keratoconus patients who had PKP. After computerized videokeratoscopic analysis, patients were randomly assigned to a test or control group with matching for midperipheral corneal keratoscopic criteria. One surgeon performed all grafts using the same technique (except for donor button punching with an 8.00 or 8.25 mm blade, depending on preoperative keratoscopy) with suction trephination (8.00) and a running 10-0 nylon suture. The PAL (total axial length minus the distance from the anterior corneal surface to the anterior lens surface) was measured by applanation ultrasonography. Refraction and videokeratoscopic analysis were done 18 months postoperatively (6 months after suture removal). RESULTS: The mean deviation from emmetropia corrected for PAL in test group patients who had trephine sizing based on the hypothesis that preoperative videokeratoscopy is a useful determining factor was 1.12 diopters (D) +/- 0.74 (SD), which was significantly smaller (P = .005) than that in the control group (2.19 +/- 0.85 D). The test group had uncorrected visual acuities of 20/50 or better. Postoperative spherical equivalent was affected by PAL (P = .0001), preoperative keratoscopy (P = .0001), and trephine disparity (P = .01). Central corneal power after grafting was influenced by keratoscopy (P = .0001) and trephine disparity (P = .002). Uncorrected visual acuity was affected by PAL (P = .001) and keratoscopic data (P = .01). CONCLUSIONS: Parameters for reducing ametropia after grafting of keratoconus patients can be developed for each surgeon based on trephine disparity dependent on preoperative keratoscopic values of the recipient midperipheral cornea and PAL. If the PAL is between 19.0 and 21.0 mm, preoperative midperipheral corneal videokeratoscopy to choose same-size or 0.25 mm different donor and recipient trephine blades is useful to achieve refractive results approximating emmetropia with the described technique.

Scanning slit delivery system.

We describe the structure and function of a delivery system designed for use with an argon fluoride excimer laser for corneal surgery. The basic principle of the delivery system is to shape the laser beam with a slit mask to a defined configuration. The image of the slit is moved across the cornea in a mathematically defined pattern to ablate a given amount of tissue, creating a new anterior radius of curvature for the cornea. The location of the beam on the cornea is determined by a rotating dove prism and a translating spherical lens, each of which is controlled by computer-regulated servomotors. The system is versatile and can be used for laser myopic or hyperopic keratomileusis, for correction of astigmatism by surface ablation, for creation of radial or transverse linear cuts, for excision of corneal buttons, and for uniform lamellar corneal smoothing procedures. The delivery system is currently used in the laboratory for the ablation of nonhuman primate corneas.

Ultrastructural study of the posterior cornea of the dogfish "Scyliorhinus canicula L".

The cornea of Elasmobranchs remains transparent and exhibits a remarkable resistance against swelling when deprived of its endothelium. Sutural fibers, which traverse the corneal stroma anteroposteriorly, may be responsible for this property. We studied the structure of the cornea of Scyliorhinus canicula L., a galeiform Elasmobranch, with special emphasis on the presence of a corneal endothelium and on its appearance during development. Observations were made with the aid of several microscopic techniques: light and specular microscopy, transmission and scanning electron microscopy. We found that the posterior corneal surface was always covered by a fibrillar material. A structural counterpart of endothelial-like cells on the posterior cornea or in the anterior chamber was never observed in adult fishes and in embryos of different stages. In contrast, in Torpedo ocellata, a torpediniform Elasmobranch, endothelial cells were seen. The significance of these findings is discussed in context with the possibility of technical artefacts and compared with other studies.