[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).

Avaliação da atividade larvicida de saponinas triterpênicas isoladas de Pentaclethra macroloba (Willd.) Kuntze (Fabaceae) e Cordia piauhiensis Fresen (Boraginaceae) sobre Aedes aegypti / Evaluation of larvicidal activity of triterpenoid saponins isolated of Pentaclethra macroloba (Willd.) Kuntze (Fabaceae) and Cordia piauhiensis Fresen (Boraginaceae) against Aedes aegypti

A atividade larvicida de quatro saponinas monodesmosídicas (1-4) isoladas de Pentaclethra macroloba e de uma saponina bidesmosídica (5) isolada de Cordia piauhiensis foi avaliada sobre larvas de estágio 3 de Aedes aegypti. As larvas foram expostas a várias concentrações (500, 250, 100, 50, 25 e 12,5 mg/mL) das saponinas por um período de 24 h. Os resultados indicam que somente as saponinas 1-3 mostraram alta atividade larvicida, com CL50 variando de 18,6 a 27,9 mg/mL. Estes resultados ressaltam as potencialidades destas saponinas como possíveis agentes larvicidas naturais.

The larvicidal activity of the four monodesmoside saponins (1-4) isolated from Pentaclethra macroloba and one bidesmoside saponin (5) from Cordia piauhiensis was evaluated on 3rd instar larvae of Aedes aegypti. The larvae were exposed to serial concentrations (500, 250, 100, 50, 25 and 12.5 mg/mL) saponins for a period of 24 h. The results indicate that, only the saponins 1-3 showed high larvicidal activity, with LC50 ranging of 18,6-27,9 mg/mL. These results suggest that these can be used as natural larvicidal agents.

[Femtosecond laser: a micromachining system for corneal surgery]. / Laser femtoseconde: système de micro-usinage pour chirurgie de la cornée.

INTRODUCTION: The authors present the diode-pumped, all-solid state, neodymium:glass femtosecond laser from the Laboratory of Ocular Biotechnology, Hotel-Dieu Hospital. MATERIALS AND METHODS: We worked with a 1,065-nm wavelength infrared laser. This laser is composed of an oscillator and amplification glass matrix mixed with neodymium. Its stretching and compression system is capable of producing pulses lasting a few hundred femtoseconds. The repetition rate is adjustable, ranging from 1 to 10 kHz, and can reach energies up to 60 microJ. The delivery system was set up on an optical table, with human corneal samples fixed to an anterior chamber system, which can be moved over the X-Y-Z axis by a computer-guided translation motor with micrometric precision. We analyzed the biological effects of laser impacts in human corneal tissue, obtained from the French Eye Bank. RESULTS: The femtosecond laser provides automated corneal cutting with a high level of precision, which can be verified on the corneal surface regularity by scanning electron microscopy analysis. Silicon samples can also be cut and can be used for calibration testing of the laser. CONCLUSION: The set-up composed of the femtosecond laser and the described delivery system enable precise corneal cutting and offer the opportunity to study its characteristics.

[BIG-H3 protein: mutation of codon 124 and corneal amyloidosis]. / Protéine BIGH3: mutation du codon 124 et amylose cornéenne.

In 1997, a group of hereditary corneal dystrophies was related to mutations in the TGFBI (BIGH3) gene. Within this group, some corneal dystrophies present particular biochemical features in that they are characterized by corneal amyloid deposition. Contrary to clinical and genetic knowledge, the biochemical characteristics of the encoded protein (Big-h3) and the mechanisms of its amyloid conversion remain unclear. We review the current knowledge on the Big-h3 protein and focus on the behavior of the codon 124 region. We discuss this protein's mechanisms of amyloid conversion from our results and previous reports as well as from other types of amyloidosis. These data provide a better understanding of the putative processes leading to the phenotypic variations linked with their respective codon 124 mutation.

[Corneal burns and matrix metalloproteinases (MMP-2 and -9): the effects of human amniotic membrane transplantation]. / Brûlures cornéennes et métalloprotéases: influence des greffes de membranes amniotiques.

PURPOSE: Human amniotic membranes have recently been used in ophthalmology to restore deleted ocular surface after burns. Matrix metalloproteinases-2 and -9 have been implicated in the development of neovascularization. In this study, MMP-2 and MMP-9 expression was analyzed by in situ zymography on rabbit corneal chemical burns with and without human amniotic membrane graft. METHOD: Corneal neovascularization was induced in 10 Fauve de Bourgogne rabbits by means of a heptanol chemical burn on controlled deep keratotomy using a Chiron ALK-E corneal shaper. Half of rabbits received acute amniotic membrane transplantation 30mn after chemical burn; the remaining five rabbits received medical treatment. In situ zymography is a recent nondestructive technique which preserved the fine morphological details of the cornea and showed the active enzyme location in different corneal layers. The MMP-2 and -9 substrate was gelatin, which was detected by fluorescent microscopy. RESULTS: There was an overexpression of MMP-2 and -9 in corneal burns versus control corneas. Expression of MMP-2 and -9 was low in corneal burn without amniotic membrane graft. Following amniotic membrane transplantation, MMP-2 and -9 were strongly expressed and clinical neovascularization and inflammation decreased. Active enzymes were located in epithelium layers in the uncovered group. In the covered group, the active enzymes were located in the anterior and posterior stromal layers. CONCLUSION: The results support a role for MMP-2 and MMP-9 in corneal burn neovascularization. Amniotic membrane transplantation can play a protective role by up-regulation of their biological expression.

Nineteen years of penetrating keratoplasty in the Hotel-Dieu Hospital in Paris.

PURPOSE: To identify changing trends in penetrating keratoplasties (PKs) performed at the Hotel-Dieu Hospital in Paris between January 1980 and December 1999 and to explain the reasons for the changes. METHODS: We retrospectively reviewed 3,736 of the 3,836 PKs performed between January 1, 1980, and December 31, 1999, and classified them into diagnostic categories. RESULTS: The most common indications for PK were keratoconus (28.8%), herpetic infections (10.9%), graft failures (9.9%), aphakic and pseudophakic corneal edema (9.9%), Fuchs' endothelial dystrophy (9.4%), and nonherpetic leucoma (7.7%). Other indications represented 23.4% of the cases. The incidence of aphakic and pseudophakic corneal edema progressively increased between 1980 and 1991, became the most frequent indication in 1991 (21.4%), and then progressively decreased. The annual number of PKs increased between 1980 and 1986, decreased between 1987 and 1997, and increased again after September 1997. The decrease was caused by both a shortage of corneal buttons, and, in 1987, the fear of transmitting diseases through corneal transplantation, particularly human immunodeficiency virus. Beginning in 1992, decreases were also associated with stringent governmental regulations of eye bank tissue. CONCLUSION: Changes in the incidence and management of corneal disorders were the primary factors leading to modifications of grafting until 1987. After 1987, corneal button shortage probably corresponded to the acquired immune deficiency syndrome epidemic. Governmental regulations of eye banking led to a severe corneal button shortage between 1992 and 1997. Despite an increase in the number of PKs performed after 1997, corneal buttons are still preferentially allocated to patients in whom there is a high probability of graft success.

Mineralocorticoid hormone signaling regulates the 'epithelial sodium channel' in fibroblasts from human cornea.

We investigated the regulation of sodium absorption by steroid hormones in embryologically diverse cells from the human eye. A cell extract from human corneal fibroblasts was positive for both the epithelial sodium channel (ENaC) and the mineralocorticoid receptor (MCR) as 82- to 85-kD and 102-kD bands, respectively, by the Western blot technique. In fluorescent, confocal and electron microscopy, the MCR was revealed as a nucleocytoplasmic protein, whereas the ENaC was almost exclusively membrane bound; both appeared aligned along actin filaments of corneal keratocytes, and both were widely colocalized in various cell types of human cornea in situ. Following reverse transcription and amplification of total RNA isolated from corneal fibroblasts, the ENaC and MCR genes in the PCR product were evident as predicted bands of 520 and 843 bp, respectively, whose sequence exhibited 100% identity with those from known human sources. The multiplication of corneal fibroblasts was influenced by both the MCR-specific antagonist RU 26752 and the natural hormone aldosterone, and these steroids also stimulated protein phosphorylation. In quantitative PCR, both the basal and aldosterone-induced levels of ENaC were diminished by the MCR-specific antagonist ZK 91587. Consequently, the ocular sodium channel appears to be regulated by steroid signalling in cells of diverse embryological origins, contrary to the existing notions where (a) this process would be limited exclusively to the epithelial cells and (b) ocular sodium transport would be regulated via the Na(+)-K(+)-ATPase in the basolateral membrane.

[Avellino dystrophy. Current diagnostic criteria]. / La dystrophie d'Avellino. Critères diagnostiques modernes.

We report a French family suffering from an Avellino corneal dystrophy diagnosed by using clinical, histological, ultrastructural and genetics findings. Our results indicate that direct corneal examination and routine histological examinations must always be associated with an assay for BIGH3 gene mutations to establish a modern and unambiguous diagnosis of a corneal dystrophy.

The intrastromal corneal ring in clinical refractive surgery: reference to results in rabbit eyes.

BACKGROUND: The intrastromal corneal ring (ISR) is a refractive device recently introduced for clinical application that is implanted in the mid-peripheral corneal stroma in order to correct myopia without invasion of the central optical zone. First clinical results of intracorneal ring segment implantation were published recently. These results reveal striking similarities to our own experimental data, only briefly published up to now. The aim of this study is to present the refractive and histopathological data of ISR implants in rabbits and to compare these results with the clinical data actually available. METHODS: Expansion/constriction effects were evaluated with a ring of constant size (7.5 x 0.5 x 0.2 mm) in channels of 7.0, 7.5, or 8.0 mm in diameter, volume effects by implantation of 7.5-mm rings with varying thickness (0.2, 0.3, 0.4 mm) into a channel of 7.5 mm, respectively. Refractive power was measured preoperatively and at day (D) 7, D14, D30 for the first and the second experiment, plus D60, D90 for the second experiment. Histological evaluations of the induced morphological changes were additionally performed at all time intervals. RESULTS: Significant (P<0.05) flattening of the cornea was obtained in all but the first (constant ring, 7.0-mm channel) implants postoperatively at D7 and/or D14, with mean dioptric changes up to -5.03+/-2.92 compared with controls. However, from D30 on, there was no statistically significant difference between operated and control eyes. Biomicroscopy and histology of the implanted eyes revealed good biocompatibility, with only rare major complications such as stromal abscess or massive neovascularization. CONCLUSIONS: Although our implantation technique differs slightly from that employed in the recent FDA studies, our results tend to confirm the maximal achievable refractive change of about -5 dpt with this procedure. Furthermore, this study is the first to demonstrate that in rabbits, ISR implantation has only a shortterm effect on refractive power. Our results indicate that long-term refractive follow-up may be necessary in human eyes prior to the introduction of ISRs as a routine procedure in refractive surgery.

Clinical, histologic, and ultrastructural features of the corneal dystrophy caused by the R124L mutation of the BIGH3 gene.

OBJECTIVE: This study was designed to describe the clinical, histologic, and ultrastructural features of the corneal dystrophy associated with the R124L mutation of the BIGH3 gene. DESIGN: Retrospective clinical and histologic review of a new genetic mutation. PARTICIPANTS: Thirty-four patients from five unrelated French families with corneal dystrophy caused by the R124L mutation of the BIGH3 gene were studied at the clinical, histologic, and ultrastructural levels. Records of patients carrying this mutation were compared with those from three unrelated patients with corneal dystrophy of Bowman's layer (CDB) type 2 (R555Q mutation) and from three unrelated patients with classic corneal granular dystrophy (R555W mutation). INTERVENTION: The mutational genetic status of the BIGH3 gene was determined for each patient, and the histologic and ultrastructural data available after corneal graft were analyzed. MAIN OUTCOMES MEASURES: Genomic DNA was extracted from peripheral blood leukocytes. Exons 4 and 12 of the BIGH3 gene were amplified by the polymerase chain reaction (PCR), and the PCR products were directly sequenced. RESULTS: All 34 patients with the R124L mutation displayed the clinical, histologic, and electron microscopic features of the dystrophy previously described as a superficial variant of corneal granular dystrophy. Combining molecular genetics with clinical and histologic findings established a clear distinction between the R555Q and R555W dystrophies. CONCLUSIONS: The R124L mutation of the BIGH3 gene is associated with specific clinical and morphologic criteria. This indicates that molecular studies are needed for an adequate classification of corneal dystrophies. All criteria are presently available to segregate the dystrophy caused by the R124L mutation (known as CDB1) from the dystrophy caused by the R555Q mutation (known as CDB2).

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.

Novel synthetic meshwork for glaucoma treatment. I. Design and preliminary in vitro and in vivo evaluation of various expanded poly(tetrafluoroethylene) materials.

A novel drainage implant for glaucoma filtering surgery (MESH) is proposed. After various expanded poly(tetrafluoroethylene) (e-PFTE) materials were evaluated, the feasibility and the short-term safety of the technique were assessed in this first pilot study in the rabbit. The porous structure and the in vitro resistance to aqueous flow of seven different e-PTFE membranes (5-80 microm average pore size) were compared. Eight Dutch pigmented rabbits were implanted with the T-shaped MESH implants made from either 20- or 50-microm average pore size e-PTFE membranes. Clinical examination, intraocular pressure (IOP) measurements, and histology analyses were performed over a period of 3 months. The contralateral nonoperated eyes served as controls. MESH implantation took less than 7 min. No postoperative hypotony, migration, or extrusion of the implant and no intraocular inflammation or infection occurred. A significant IOP reduction in the implanted eyes was obtained past postoperative day 21 with the 20-microm material implant. The drainage efficacy was correlated with the degree of colonization of the porous materials and the inner spacing of the implant as observed by histology. With a filtering patency 3 times longer than conventional trabeculectomy and laser sclerectomy, MESH surgery is a promising technique for glaucoma treatment. Further studies are underway to enhance the device efficacy and understand the mechanism of filtration.

Scleral and episcleral histological changes related to encircling explants in 20 eyes.

PURPOSE: To investigate scleral and episcleral histological alterations induced by encircling explants used in scleral buckling procedures. METHODS: We performed a histopathological study of 20 enucleated eyes after failure of retinal detachment surgery including encircling scleral buckle. RESULTS: Nonabsorbable materials were encapsulated and often gave rise to a limited scleral invagination. The inner capsular surface was regular in 10 silicone explants, it was partially covered with hydrogel fragments and a granulomatous foreign body giant cell reaction in 8 hydrogel explants. One Arruga thread was encased in fibrosis and a catgut circle showed no encapsulation. Other changes were mostly related to the long-standing retinal detachment: peripheral anterior synechiae, anterior uveal effusion, persistent retinal detachment, retinal gliosis, retinal atrophy, retinal breaks, and silicone oil droplets. CONCLUSION: All nonabsorbable explants underwent encapsulation and prompted scleral invagination. A granulomatous reaction accompanied hydrogel fragmentation. The long term fragmentation impact on implanted eyes remains unknown.

[Phototherapeutic keratectomy in the treatment of Groenouw's type I corneal dystrophy]. / La photokératectomie thérapeutique dans le traitement de la dystrophie de Groenouw I.

PURPOSE: This study was designed to investigate the therapeutic potential of phototherapeutic keratectomy (PTK) for the treatment of corneal granular dystrophy. PATIENTS AND METHODS: PTK was performed with the Excimed UV 200, (Summit Technology, Inc) on a series of 27 eyes of 22 patients with corneal granular dystrophy. Mean patients' age was 34.6 years. The mean follow-up period was 31 months (ranged from 6 to 52 months). The changes in best corrected visual acuity and spherical equivalent were evaluated at 6, 12, 18 and 24 months. RESULTS: All of patients achieved visual improvement. Mean preoperative best corrected visual acuity (BCVA) was 20/100 and mean postoperative BCVA was 20/30 at one year. The mean hyperopic shift caused by tissue ablation was +/- 2.8 D after one year. CONCLUSION: Corneal grafting was the standard treatment for visually disabling granular dystrophy, but PTK has significant advantages over this procedure and must now be the standard method of managing corneal granular dystrophy when intervention is required.

Exophiala (Wangiella) dermatitidis keratitis after keratoplasty.

We report the first French case of an intraocular infection due to Exophiala (Wangiella) dermatitidis. Two months after a second corneal transplant for congenital hereditary endothelial dystrophy, the patient presented with ocular pain and corneal infiltrates leading to the graft rejection. Diagnosis was established by positive direct examination and cultures of the same fungus from corneal buttons, iris biopsies and ablated sutures.