The smooth optical surface of the cornea is maintained by a tear film, which adheres to a variety of microprojections. These microprojections increase the cell surface area and are thought to improve the movement of oxygen, nutrients, and metabolic products across the outer cell membranes. However, little is known of these structural adaptations in vertebrates inhabiting different environments. This field emission scanning electron microscopic study examined the cell density and surface structure of corneal epithelial cells across 51 representative species of all vertebrate classes from a large range of habitats (aquatic, amphibious, terrestrial, and aerial). In particular, we wished to extend the range of vertebrates to include agnathans and some uniquely Australian species, such as the Australian lungfish (Neoceratodus forsteri), the Australian galah (Eolophus roseicapillus), the Australian koala (Phascolarctos cinereus), and the rat-tailed dunnart (Sminthopsis crassicaudata). Epithelial cell densities ranged from 28,860 +/- 9,214 cells mm(-2) in the flathead sole Hippoglossoides elassodon (a marine teleost) to 2,126 +/- 713 cells mm(-2) in the Australian koala (a terrestrial mammal), which may indicate a reduction in osmotic stress across the corneal surface. A similar reduction in cell density occurred from marine to estuarine to freshwater species. The structure and occurrence of microholes, microplicae, microridges, and microvilli are also described with respect to the demands placed on the cornea in different environments. All species that spend significant periods out of an aquatic environment possess microvilli and/or microplicae. These include all of our species of Mammalia, Aves, Reptilia, Amphibia, and even one species of Teleostei (Australian lungfish). Well-developed microridges occur only in teleosts in high osmolarity environments such as marine or estuarine habitats. Clear interspecific differences in corneal surface structure suggest a degree of adaptive plasticity, in addition to some phylogenetic trends.

Marked deficiency of muscle adhalin, a 50 kDa sarcolemmal dystrophin-associated glycoprotein, has been reported in severe childhood autosomal recessive muscular dystrophy (SCARMD). This is a Duchenne-like disease affecting both males and females first described in Tunisian families. Adhalin deficiency has been found in SCARMD patients from North Africa Europe, Brazil, Japan and North America (SLR & KPC, unpublished data). The disease was initially linked to an unidentified gene on chromosome 13 in families from North Africa, and to the adhalin gene itself on chromosome 17q in one French family in which missense mutations were identified. Thus there are two kinds of myopathies with adhalin deficiency: one with a primary defect of adhalin (primary adhalinopathies), and one in which absence of adhalin is secondary to a separate gene defect on chromosome 13. We have examined the importance of primary adhalinopathies among myopathies with adhalin deficiency, and describe several additional mutations (null and missense) in the adhalin gene in 10 new families from Europe and North Africa. Disease severity varies in age of onset and rate of progression, and patients with null mutations are the most severely affected.

X-linked recessive Duchenne muscular dystrophy (DMD) is caused by the absence of dystrophin, a membrane cytoskeletal protein. Dystrophin is associated with a large oligomeric complex of sarcolemmal glycoprotein. The dystrophin-glycoprotein complex has been proposed to span the sarcolemma to provide a link between the subsarcolemmal cytoskeleton and the extracellular matrix component, laminin. In DMD, the absence of dystrophin leads to a large reduction in all of the dystrophin-associated protein. We have investigated the possibility that a deficiency of a dystrophin-associated protein could be the cause of severe childhood autosomal recessive muscular dystrophy (SCARMD) with a DMD-like phenotype. Here we report the specific deficiency of the 50K dystrophin-associated glycoprotein (M(r) 50,000) in sarcolemma of SCARMD patients. Therefore, the loss of this glycoprotein is a common denominator of the pathological process leading to muscle cell necrosis in two forms of muscular dystrophy, DMD and SCARMD.

Primary adhalin (or alpha-sarcoglycan) deficiency due to a defect of the adhalin gene localized on chromosome 17q21 causes an autosomal recessive myopathy. We evaluated 20 patients from 15 families (12 from Europe and three from North Africa) with a primary adhalin deficiency with two objectives: characterization of the clinical phenotype and analysis of the correlation with the level of adhalin expression and the type of gene mutation. Age at onset and severity of the myopathy were heterogeneous: six patients were wheel-chair bound before 15 years of age, whereas five other patients had mild disease with preserved ambulation in adulthood. The clinical pattern was similar in all the patients with symmetric characteristic involvement of trunk and limb muscles, calf hypertrophy, and absence of cardiac dysfunction. Immunofluorescence and immunoblot studies of muscle biopsy specimens showed a large variation in the expression of adhalin. The degree of adhalin deficiency was fairly correlated with the clinical severity. There were 15 different mutations (10 missense, five null). Double null mutations (three patients) were associated with severe myopathy, but in the other cases (null/missense and double missense) there was a large variation in the severity of the disease.

Several studies have demonstrated the feasibility of gene cardium. Transgene expression is predominant in the left transfer into the heart muscle. However, all the available ventricle and the interventricular septum but limited in the data also indicate that the extent of transfection remains right ventricle. In vivo echocardiographic measurements of limited. As an alternative method to intravascular administhe left ventricular diameters at end diastolic and end systration, we have developed a novel strategy which uses tolic times show no difference between virus-and shamthe pericardial sac. When a replication-deficient adenovirus injected animals, thus indicating a good clinical tolerance containing the cDNA encoding a bacterial ␤-galactosidase to this strategy of virus delivery. The same protocol has is injected into the pericardial sac of adult Wistar rats the been used with the same efficiency in mice, which leads staining is exclusively restricted to the pericardial cell layus to propose injection into the pericardial sac as an effecers. However, injecting a mixture of collagenase and hyalutive and harmless method for gene transfer into the heart ronidase together with the virus, leads to a large diffusion muscle. of the transgene activity, reaching up to 40% of the myo-