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We have compared clinical (Schirmer I test, BUT and rose bengal staining), laboratory (lysozyme and lactoferrin tear levels) and histological tests (impression cytology) in 165 eyes from 85 patients with primary Sjögren's syndrome and in 80 eyes from 40 control subjects. Impression cytology can provide the location of cellular alterations on the ocular surface and information on the severity of the disease. The upper bulbar and interpalpebral areas from patients with primary Sjögren's syndrome were shown to have cellular alterations early in the course of the disease, while the lower bulbar and lower palpebral areas were only affected in severe cases. Statistical analysis has shown that impression cytology and rose bengal staining are the most specific and sensitive methods for the diagnosis of primary Sjögren's syndrome.

A phytoene dehydrogenase-deficient mutant of Mucor circinelloides accumulating only phytoene was transformed with the gene encoding the corresponding enzyme (carB gene) of Phycomyces blakesleeanus. Carotenoids derived from phytoene were detected in the transformants showing that the P. blakesleeanus carB gene complements the M. circinelloides carB mutation. These newly formed carotenoids accumulated in low quantities, indicating that functional complementation was poor. carB mRNA molecules correctly transcribed were detected in the transformants, but they represented a small proportion of the total population of carB-derived mRNAs, mostly constituted by truncated transcripts and by transcripts longer than the transcript that is functional in Phycomyces. These results showed that the P. blakesleeanus carB gene was expressed in M. circinelloides and suggested that the poor complementation observed was owing, at least in part, to the lack of specificity in the recognition of the transcription initiation and termination signals of the P. blakesleeanus carB gene by the M. circinelloides transcriptional machinery.

Purpose The aim of this work was to evaluate ultrastructural alterations of the conjunctiva during the clinical course of keratoconjunctivitis sicca (KCS), and to detect its earliest and most characteristic morphological changes. Methods The conjunctiva was studied in biopsies from 75 patients and 10 controls. Patients were classified according to the results of the Schirmer I test, break-up time, rose Bengal staining, osmolarity and impression cytology. Results The conjunctiva in these KCS patients showed progressive hyperplasia, hypertrophy and cellular flattening, with diminution of goblet cell density and microvilli. In the severe cases, the epithelial cells lost their organelles, and fibrous material increased. From the early phases of KCS, clear nuclear alterations (indentation, binucleation) were found, but pyknotic nuclei or anucleated cells were only observed in the most severe cases. From the earliest stages to the most severe cases of KCS, decreases in cell membrane interdigitations were observed parallel to increases in the number and size of desmosomes. There were also increases in the number of inflammatory cells. Alterations in blood vessels were only observed in the most severe cases. Conclusions Morphological studies alone were able even in the earliest phases of KCS, to detect the squamous metaplasia that progresses from the surface of the epithelium to the connective tissue. This degenerative or adaptative cellular process was characterized mainly by marked proliferation of the cytoskeleton and a general loss of organelles, mitochondria being the least affected.