Immunoassay of human tear lysozyme.

We used a single radial immunodiffusion technique to determine the lysozyme content of human tears. The level in healthy subjects with this method was 1.3 mg/ml of tears. In patients with trachoma, the level was low, 0.9 mg/ml of tears. Detecting the reduction in lysozyme level in patients with trachoma proved the utility of this method.

Expression of gelatinase B in trachomatous conjunctivitis.

BACKGROUND/AIMS: Gelatinase B is a matrix metalloproteinase involved in extracellular matrix (ECM) breakdown often associated with scarring and other pathological disorders. It was investigated whether gelatinase B is involved in the pathogenesis of ECM degradation associated with trachomatous conjunctivitis. METHODS: Conjunctival biopsy specimens obtained from six patients with active trachoma, six patients with active vernal keratoconjunctivitis (VKC), and seven control subjects were studied. Immunohistochemical techniques and a specific monoclonal antibody against human gelatinase B were used, and a monoclonal antibody against macrophage CD68 to identify mononuclear cells with gelatinase B immunoreactivity. In addition, quantitative zymography was used to compare the activity of gelatinase B in conjunctival biopsy specimens from seven patients with active trachoma and seven control subjects. RESULTS: Gelatinase B was detected by immunohistochemistry only in polymorphonuclear cells located in the vascular lumens in three normal conjunctival biopsy specimens. In all trachoma specimens and in five VKC specimens, gelatinase B was localised in monocyte/macrophage cells, positive for the CD68 marker, and in polymorphonuclear cells. The majority of the latter cell type was located in intravascular spaces. Compared with VKC specimens, trachoma specimens showed significantly more immunoreactive gelatinase B monocyte/macrophage cells (52.3 (21.9) v 8.2 (6.4); p <0.001) and polymorphonuclear cells (23.2 (14.2) v 6.3 (5.4); p = 0. 013). Activated macrophages with giant cell morphology clearly stained with the gelatinase B specific monoclonal antibody were observed in trachoma specimens. Zymography revealed that gelatinase B levels in trachoma specimens were significantly higher than the levels found in normal conjunctiva (1739.6 (1078.3) v 609.3 (395.9) scanning units; p = 0.0127). CONCLUSIONS: The increased activity of gelatinase B and numbers of inflammatory cells containing gelatinase B in trachoma specimens suggest that this enzyme plays a part in the pathogenesis of conjunctival scarring in trachoma.

[Tear malate dehydrogenase, lactate dehydrogenase, and their isoenzymes in normal Chinese subjects and patients of ocular surface disorders].

Levels of malate dehydrogenase (MDH), lactate dehydrogenase (LDH) and their isoenzymes in tears of normal Chinese subjects and patients with ocular surface disorders were determined. The normal values of tear LDH and MDH were found to be 45.51 +/- 23.00-81.35 +/- 37.84 mumol.s-1/L and 11.00 +/- 5.33-19.50 +/- 9.17 mumol.s-1/L respectively, disregarding sex or eye distinction. The LDH/MDH ratio reflected sensitively the metabolism of corneal and conjunctival epithelium. The MDH isoenzymes comprised MDHs and MDHm, the former accounting for 80.0%-89.1%. The LDH isoenzymes comprised 5 varieties, of which the ratio H/M of subunit H to subunit M was 0.196 +/- 0.02. The changes in LDH isoenzymes were helpful to the differential diagnosis of external eye diseases, and the increase of MDHm reflected sensitively the degree of injury to the corneal epithelium.

Tear malate dehydrogenase, lactate dehydrogenase and their isoenzymes in normal Chinese subjects and patients of ocular surface disorders.

PURPOSE: To determine levels of malate dehydrogenase (MDH), lactate dehydrogenase (LDH) and their isoenzymes in tears of normal Chinese subjects and patients with ocular surface disorders. METHODS: The age range of normal subjects was 10-88, with 136 male and 128 female subjects. 123 patients suffered from ocular surface disorders. Tears were collected from lower fornix on Xinghua filter disc (0.1mm thick, 5mm in diameter). The values of tear MDH and LDH were determined by MONARCH-2000 Analyzer (U. S. A.). Their isoenzymes were separated by acetate cellulose electrophoresis and were determined by Model CDS-200 light densitometer. RESULTS: The normal values of tear LDH and MDH were 45.51 + 23.00-81.35 + 37.84 umol.s-1/L and 11.00 + 5.33-19.50 + 9.17 umol.s-1/L respectively, disregarding sex or eye distriction (P > 0.05). The values of tear LDH and MDH in the group aged 10-19 were significantly lower than in another groups (P < 0.05). 95% normal ranges of tear MDH aged below 19 and above 20 were 3.63-19.90 umol.s-1/L and 4.20-36.64 umol s-1/L. 95% normal ranges of tear LDH aged below 19 and above 20 were 17.69-82.93 umol.s-1/L and 21.47-150.41 umol.s-1/L. The MDH isoenzymes comprised MDHs and MDHm, the former accounting for 80.0-89.1%. The LDH isoenzymes comprised 5 varieties, of which the ratio H/M of subunit H to subunit M was 0.196 + 0.02. Levels of tear LDH,MDH and their isoenzymes in different diseases were various. CONCLUSIONS: Tear LDH/MDH ratio reflected sensitively the metabolism of corneae and conjunctival epithelium. The changes in LDH isoenzymes were helpful to the differential diagnosis of external eye diseases, and the increase of MDHm reflected sensitively the degree of injury to the corneal epithelium.

The significance of tears' lactate dehydrogenase in health, and external eye diseases.

The normal values of the lactate dehydrogenase contents of tears in the presence of pyruvate and lactate as substrates were determined. The quotients of the LDH values measured in the presence of the two different substrates, and also the ratio of the subunits (H and M) of the isoenzymes, differ from the corresponding data of blood serum. In the event of external inflammatory diseases of the eye, LDHp/LDHl decreases in the tears; this deviation is enhanced by treatment with drugs inhibiting protein synthesis, primarily iododeoxyuridine. Conversely, in trachoma the ratio of the LDH isoenzymes containing mainly M (muscle) subunits are increasing. In diabetes mellitus the shift of metabolism toward the citrate cycle is reflected to a larger extent by the LDH values in the blood serum than by those in tears.