Glia, cytokines, and neurotoxicity

Activated glial cells (microglia and astrocytes) are a hallmark of several neurodegenerative disorders. A growing body of evidence supports the hypothesis that activation of glial cells by cytokines contributes to neurotoxicity. Although the precise mechanisms underlying glia-mediated neurotoxicity are unclear, it has been proposed that the generation of toxic free radicals or other neurotoxins is involved. This review focuses on the role of immune mediators released by activated glial cells in causing or preventing neuronal injury. Also, techniques used to assess neurotoxicity are discussed. It is hoped that research in this field will yield insights that will result in new therapies for neurodegenerative disorders.     

Attitudes of Dublin accident and emergency department doctors and nurses towards the services offered by local general practitioners

We report a case of a depressed patient who received a full course of electroconvulsive therapy (ECT) 2 weeks post four-vessel coronary artery bypass graft surgery (CABG). ECT was well tolerated, and a full remission was induced. In spite of the lack of information in the literature concerning guidelines for administering ECT to a patient with recent postoperative status, we were encouraged by this patient's response. This case suggests that ECT may be considered a viable treatment option for refractory depression associated with severe medical comorbidity and recent post-operative status such as in CABG.     

Cleavage of Müllerian inhibiting substance activates antiproliferative effects in vivo

Müllerian inhibiting substance (MIS), an inhibitor of growth and development of the female reproductive ducts in male fetuses, requires precise proteolytic cleavage to yield its biologically active species. Human plasmin is now used to cleave and, thereby, activate immunoaffinity-purified recombinant human MIS at its monobasic arginine-serine site at residues 427-428. To avoid the need for exogenous enzymatic cleavage and to simplify purification, we created an arginine-arginine dibasic cleavage site (MIS RR) using site-directed mutagenesis to change the serine at position 428 (AGC) to an arginine (cGC). The mutant cDNA was then stably transfected into a MIS-responsive ocular melanoma cell line, OM431, followed by cloning for amplified expression to test its biological activity in vitro and in vivo. Media from each clone were assayed for production of MIS RR by a sensitive ELISA for holo-MIS, and high- and low-producing clones were selected for further study. Media from the highest MIS RR producer caused Müllerian duct regression in an organ culture bioassay. Other transfections were done with an empty vector (pcDNAI Neo) or a construct lacking the leader sequence and thus failing to secrete MIS, to serve as controls. The OM431 clones containing the MIS RR mutant were growth inhibited in monolayer culture. The high- and low-producing MIS RR OM431 clones, along with transfected OM431 controls, were injected into the tail veins of immunosuppressed severe combined immunodeficiency mice for in vivo analyses. Four to 6 weeks later, pulmonary metastases were counted in uniformly inflated lungs. OM431 clones containing the more easily cleaved MIS RR displayed a significant dose-dependent reduction in pulmonary metastases when compared to the lungs of animals given injections of OM431 clones containing empty vector, leaderless MIS, or wild-type MIS that requires activation by plasmin cleavage. Since the purification protocol of MIS RR is less complicated than that for wild-type MIS, which requires subsequent enzymatic activation, MIS RR can be used for scale-up production with increased yields for further therapeutic trials against MIS-sensitive tumors.     

Corneal epithelial recovery following photorefractive keratectomy

AIMS: To further understand the morphological and functional recovery of corneal epithelium following excimer laser photorefractive keratectomy (PRK). METHODS: The right eyes (group 1) of 15 male, New Zealand white rabbits weighing 2-3 kg underwent PRK. The left eye of each rabbit (group 2) underwent simple mechanical de-epithelialisation and were examined as treated controls. Both eyes of another eight rabbits (group 3) served as untreated controls. All eyes underwent a corneal epithelial permeability study by fluorophotometry at 2, 4, and 8 weeks after surgery. Five animals in groups 1 and 2 were sacrificed at 9, 10, and 12 weeks after surgery. The animals in group 3 were sacrificed at the end of the 12 week experimental period. Both eyes of each sacrificed animal were enucleated immediately and processed for both haematoxylin and eosin stain and electron microscopic study. The electron micrograph was magnified to 14,000x and the extent of hemidesmosome formation was quantified and analysed. RESULTS: The corneal epithelial barrier to sodium fluorescein was subnormal and returned to a normal barrier state 4 weeks after PRK in group 1 whereas it was normal in group 2 throughout the examination period. The extent of hemidesmosome formation was abundant yet subnormal in both groups 1 and 2 up to 12 weeks, when compared with that in group 3. CONCLUSION: The corneal epithelium regained its functional barrier 4 weeks after PRK in rabbits while the extent of hemidesmosome formation was still subnormal 12 weeks after mechanical de-epithelialisation, with or without PRK.