Changing threshold for cataract surgery in Denmark between 1980 and 1992. Results from the Danish Cataract Surgery Outcomes Study. II

In Denmark the number of cataract extractions has increased 350% from 1980 to 1991. During the same period the elderly population at risk has only increased 17%, and thus cannot account for the large increase in the number of extractions. In order to investigate whether more comprehensive clinical indications could be a possible explanation, we compared pre-operative visual acuity and visual impairment in two consecutive samples of Danish cataract surgery patients obtained in 1980 (n = 73) and in 1992 (n = 290). Criteria for inclusion were similar and both samples were representative for the whole country. During the period mean pre-operative visual acuity increased from 0.04 to 0.16 in the eye enlisted for surgery (p < 0.00001). Visual functional impairment could be compared by using the same questionnaire for patient interview in 1992 as was used in 1980. In 1992 the degree of impairment was significantly less for reading, outdoor orientation and self care activities. A change in surgical threshold or clinical indications for surgery appears to be a major contributing factor to the large increase in surgical rates, even though a trend to perform second eye cataract surgery more often might also be of some importance.     

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.     

Immunological and functional characterization of Mycobacterium leprae protein antigens: an overview

A major focus of leprosy research in the last 10 years has been the identification and characterization of antigens of Mycobacterium leprae that interact with antibodies and T cells of the host's immune response. Through the combined efforts of many different laboratories, a substantial number of protein antigens have been identified and characterized. In this MicroReview we present an updated list of M. leprae protein antigens, and, with emphasis on recent developments, summarize what is known regarding their functional and immunological features.     

Malignant oncocytoma of the lacrimal sac: ultrastructure and immunocytochemistry

Malignant oncocytoma of the lacrimal sac is a rare tumor. A third example is presented here. In addition to oncocytes, tumor acini also harbored a few mucin cells and myoepithelial cells. The presence of immunoreactive lactoferrin and secretory piece suggested a resemblance to parotid acinic cell carcinomas. The difference was marked by a compact acinar structure and myoepithelial cell differentiation in the oncocytoma.     

The development and cross-validation of methods based on radioimmunoassay and LC/MS-MS for the quantification of the class III antiarrhythmic agent, MK-0499, in human plasma and urine

An analytical method based on radioimmunoassay (RIA) has been developed for the determination of the antiarrhythmic agent, MK-0499, in plasma and urine. Owing to the potency of the drug, the specificity of this assay in human plasma could not be adequately determined using conventional RIA procedures. A highly specific procedure, based on LC/MS-MS, was developed to cross-validate the RIA. The lower quantifiable limits of the RIA and LC/MS-MS-based methods were 0.05 and 0.013 ng ml-1, respectively. Cross-validation data, compared using paired student's t-test regression analysis, showed excellent correlation between methods. The mass spectrometric assay was also used to simultaneously measure plasma concentrations of unlabeled and 14C-labeled MK-0499 following administration of the drug at high specific activity to volunteers.