Amniotic membrane graft: histopathological findings in five cases.

Amniotic membrane transplantation (AMT) is an effective treatment for ocular surface reconstruction; however, the mechanisms through which amniotic membrane (AM) exerts its effects as well as its fate after transplantation have not been entirely elucidated and have been investigated only in part. We evaluate the integration of AM in the host cornea in five patients who underwent AMT as the result of Bowen's disease, band keratopathy, radio- or cryotherapy-induced keratopathy, chemical burn or post-herpetic deep corneal ulcer with descemetocele. Due to persistent opacification in four cases and a progressing tumor in one case, penetrating keratoplasty (PK) and enucleation were performed as early as 2 months and up to 20 months after AMT. The corneas were analyzed histopathologically. To evaluate AM remnants, corneas were stained with periodic acid Schiff's reaction (PAS), Alcian blue, and Gomory and Masson trichrome; immunostaining including collagens III and IV antibodies was also performed. None of the corneas showed remnants of AM. In all cases, we observed discontinuity of Bowman's membrane. In three cases, the corneal epithelium was completely restored, ranging from three to six cell layers. In the other two cases, we detected an intense inflammatory reaction with rich neovascularization; the epithelial surface of the central cornea was completely restored, while at the periphery of the cornea goblet mucus-producing cells were present. Although clinically useful in all cases, restoration of a stable corneal epithelium through AMT is limited by the extent and severity of limbal stem cell deficiency (LSCD). The lack of histologically documented AM remnants in our cases seems to explain the efficacy of AMT more through its biological properties than through its mechanical properties.

Spleen depletion in neonatal sepsis and chorioamnionitis.

Neonatal sepsis and chorioamnionitis induce morphologic modifications and shrinkage of the thymus. We show fetal and neonatal morphologic modifications of the spleen in the same autopsy subjects as previously used to describe thymus shrinkage, including 10 preterm or full-term neonates who died of proven sepsis within 48 hours after birth and 20 fetuses spontaneously aborted because of extensive ascending chorioamnionitis. Control subjects included 10 fetuses from induced termination of pregnancy and 10 neonates who died suddenly during the perinatal period without evidence of chorioamnionitis. Spleen cell populations were studied by means of immunohistochemical analysis. Neonatal sepsis occurred with severe spleen depletion, involving both B and T lymphocytes (P < .001). Fetuses with chorioamnionitis also showed spleen cell depletion. These observations, to our knowledge not described before, indicate that preterm and term neonates show an inflammatory reaction similar to that of adult patients and that severe chorioamnionitis is associated with a nonspecific inflammatory response comparable to that of sepsis.

Digital surface microscopy analysis of conjunctival pigmented lesions: a preliminary study.

The objective of this study was to investigate whether digital surface microscopy (DSM) could be used for the follow-up and comparison of malignant and benign conjunctival pigmented lesions (CPLs). Thirty-nine CPLs [16 de novo malignant melanomas (MMs), one MM arising from primary acquired melanosis (PAM), six PAMs and 16 naevi] were digitally analysed and biopsied. All of the PAMs and 10 naevi, which had not been surgically excised, were followed up using DSM. Thirty parameters were evaluated grouped into four categories: geometry, colour, texture and islands of colour. None of the CPLs that were followed up, which comprised 10 naevocytic naevi and seven PAMs, showed any morphological change at DSM analysis, except for one PAM which developed an MM 1 year later. Of the geometric variables examined, the area, maximum diameter and minimum diameter showed significantly higher values in MMs compared with benign CPLs. With regard to the colour of CPLs, MMs were significantly darker and bluer than naevi. In the texture group, contrast was significantly higher in MMs. In the islands-of-colour group, the imbalance of blue-grey regions and the presence of dark areas were significantly higher in MMs. DSM greatly simplified the follow-up of CPLs, such as PAMs with atypia, by providing satisfactory quality images with high reproducibility; this technique is also easy to use and well accepted by patients. Moreover, this preliminary study allowed us to determine which objective variables could be important for distinguishing between benign CPLs and conjunctival MMs.

Nerve growth factor expression in human dystrophic muscles.

Nerve growth factor (NGF) is a neurotrophin that is expressed during muscle development and is also capable of favoring muscle regeneration in experimental studies. The presence of NGF in muscular dystrophies, such as Duchenne and Becker muscular dystrophies, has never been fully explored. By means of immunohistochemistry, we show that regenerating muscle fibers from such patients consistently express NGF, as do myofibroblasts and mast cells. By contrast, rest fibers from dystrophic patients, as well as muscle fibers from healthy, control patients and even regenerative muscle fibers in polymyositis do not show NGF immunoreactivity. The paracrine effect of NGF on muscle regeneration, as well as its chemoattractant capacities for mast cells, may contribute to explaining why regenerating fibers most frequently occur in clusters and why mast cells are more numerous in dystrophic muscles. Moreover, being a mediator of wound healing and tissue fibrosis, NGF may contribute to long-term muscle regeneration impairment by tissue fibrosis in the muscular dystrophies.