C57BL/6 mice lacking Muc1 show no ocular surface phenotype.

PURPOSE: To test the hypothesis that a membrane-spanning mucin, Muc1, facilitates the spread of tear film and protects against bacterial adherence. METHODS: Age-matched, Muc1 null mice and wild-type mice of C57BL/6 genetic background were used for comparison. Eyes were examined by slit lamp biomicroscopy with fluorescein solution to assess epithelial damage and tear film stability. Structure of the ocular surface epithelia was examined by light microscopy, scanning and transmission electron microscopy, and wholemount confocal microscopy. Bacterial adherence assay was performed on in vivo corneas with Pseudomonas aeruginosa containing a plasmid encoding green fluorescent protein, followed by wholemount confocal microscopy. Real-time reverse transcription-polymerase chain reaction was performed using Muc4-specific primers to quantitate Muc4 mRNA expression in ocular surface tissues. RESULTS: No differences were found between Muc1 null and control mice in any parameter tested. Ocular surface epithelia of Muc1 null mice of the C57BL/6 strain had a normal appearance of surface microplicae, a well-developed glycocalyx on the apical cell membrane, and a normal appearance of goblet cell mucin packets. There was no convincing evidence that bacterial adherence on the cornea was increased in Muc1 null mice. Muc4 mRNA expression was not upregulated in Muc1 null mice compared with control. No ocular surface infections were observed in Muc1 null mice of the C57BL/6 strain (n = 204), which were housed in the animal facility over a period of 26 months. CONCLUSIONS: Muc1 null mice of C57BL/6 background appeared normal in all respects tested. These data differ from the reported phenotype in the mice of the C57BL/6 x SVJ129 background, which show development of blepharitis and conjunctivitis.

Alteration of mucin in human conjunctival epithelia in dry eye.

PURPOSE: The objective of this study was to determine whether alteration in mucins could be detected in patients with dry eye symptoms by using the monoclonal antibody H185, which recognizes carbohydrate epitopes on mucin molecules. METHODS: Immunohistochemistry and immunoelectron microscopy was used to examine binding of H185 antibody to conjunctival cells obtained by nitrocellulose filter paper stripping (impression cytology). Two study populations were examined. Study I included 22 patients with dry eye symptoms and 13 normal volunteers. Study II included 16 aqueous-deficient dry eye patients and 14 age-matched control subjects. RESULTS: Results of the studies demonstrated significant differences in binding patterns of H185 to conjunctival cells in normal eyes compared with those of patients with dry eye symptoms. In normal eyes, the antibody bound to apical cells in a mosaic pattern, with cells exhibiting either light, medium, or intense binding. A predominant pattern in patients with dry eye symptoms was loss of the mosaic pattern with replacement by a "starry sky" pattern in which there was a lack of apical cell binding (hence, dark sky) but increased binding to goblet cells (hence, stars in the sky). The starry sky pattern correlated with rose bengal staining. CONCLUSIONS: From these studies it is concluded that there is an alteration either in mucin distribution or mucin glycosylation on the surfaces of apical conjunctival cells in dry eye and that glycosylation of goblet cell mucins changes with the disease.

Actin 'purse string' filaments are anchored by E-cadherin-mediated adherens junctions at the leading edge of the epithelial wound, providing coordinated cell movement.

At the leading edge of healing embryonic epithelium, cables of actin filaments appear to extend from cell to cell, forming a ring around the wound circumference. It has been hypothesized that this actin filament cable functions as a contractile 'purse string' to facilitate wound closure. We have observed this cable in large, circular healing epithelial wounds in corneas of adult mice. To elucidate the role of the actin filament cable, we characterized the molecular components associated with the cell-cell junction where the actin filament cable inserts and with the actin filament cable itself, and we studied the effect of disruption of the cable using an E-cadherin function-blocking antibody, ECCD-1. Localization of E-cadherin and the direct association of catenins with actin filament cable at the cell-cell interface of the actin cable confirmed that the cell-cell junction associated with the actin filament cable is an adherens junction. The E-cadherin function-blocking antibody caused disruption of the actin filament cable and induction of prominent lamellipodial extensions on cells at the leading edge, leading to a ragged uneven epithelial wound margin. These data demonstrate that cell-to-cell associated E-cadherin molecules link the actin filament cable, forming a functional adherens junction, and that the actin filament cable plays a role in coordinating cell movement.

[A case of gastric cancer with peritonitis carcinomatosa and malignant biliary stenosis treated by intraperitoneal CDDP and MMC with UFT therapy].

A 55-year-old man, who complained of vomiting, was diagnosed as having a Borrmann type 3 gastric cancer (T3N3P2H0M0: Stage IVb). He was treated by distal gastrectomy. After four months, PTCD was applied for malignant biliary stenosis due to lymphnode metastasis. Since ascites due to peritonitis carcinomatosa developed about six months after operation, UFT therapy combined with CDDP 50 mg and MMC 10 mg intraperitoneally were performed. The patient became gradually less aware of subjective symptoms after chemotherapy. Three months later, he enjoyed a good general condition and all of the fixed tubes were removed. This chemotherapy seemed effective to support his quality of life.

Diagnostic usefulness and cutoff value of Schirmer's I test in the Japanese diagnostic criteria of dry eye.

BACKGROUND: Recently, the diagnostic criteria of dry eye have been tentatively approved in Japan. The aim of this study was to evaluate the diagnostic usefulness and the cutoff value of Schirmer's I test on applying these Japanese diagnostic criteria to patients with Sjögren's syndrome. METHODS: One hundred eyes of 50 patients with Sjögren's syndrome underwent a series of diagnostic tests, including Schirmer's I test, cotton thread test, tear film break-up time (BUT), fluorescein staining, and rose bengal staining. They were classified into definite dry eye, probable dry eye, and normal eye according to the Japanese criteria. The diagnostic usefulness of Schirmer's I test was evaluated in comparison with that of the cotton thread test or BUT, based on the diagnostic outcome by combination of the individual tests plus vital staining tests. The cutoff value of Schirmer's I test was evaluated, based on the results of sensitivity and specificity rates at each cutoff value from 0 to 10 mm. RESULTS: The diagnostic usefulness of Schirmer's I test was inferior to that of BUT, but superior to that of cotton thread test. The sensitivity and specificity were 80% and 53%, respectively, at a cutoff value of 5 mm and 88% and 35%, respectively, at a cutoff value of 10 mm. CONCLUSION: In investigation of dry eye, both Schirmer's I test and BUT should be performed to detect tear abnormalities. The cutoff value of 5 mm in Schirmer's I test seems to be justified provided that the purpose of the test is comparison of inter-institutional data, not screening.

Observation of precorneal tear film in patients with Sjögren's syndrome.

The present study was designed to examine whether the lipid layer of the precorneal tear film has a role in the pathogenesis of dry eye. The study involved 104 eyes of 52 patients with primary Sjögren's syndrome. The lipid layer of the precorneal tear film was observed by non-contact specular microscopy, and relationships between grade of interference color and the tear function parameters were examined. The lipid layer was classified from Grade 1, with no interference color, to Grade 4, of most intense interference color. In addition to these four grades, an oil droplet type not previously described was characterized. The grade of interference color was closely related to the intensity of vital staining. The oil droplet type was associated with more severe ocular surface damage than were Grades 1 through 4. The findings of this study suggest that the state of the lipid layer of the tear film may have some involvement in the pathogenesis of dry eye.

Ocular surface damage and tear lactoferrin in dry eye syndrome.

We studied the relationship between the severity of ocular surface damage and the level of tear lactoferrin in primary and secondary Sjögren's syndrome and keratoconjunctivitis sicca not associated with Sjögren's syndrome. A significant negative correlation was found between Rose Bengal staining score and level of tear lactoferrin in all three groups. Analysis of covariance disclosed no significant differences in regression lines for Rose Bengal staining score vs tear lactoferrin level among the three groups. The three regression lines appeared to be identical to each other. These findings indicate that the severity of ocular surface damage due to dry eye largely depends on the tear secretory function of the lacrimal gland, and that the function of the lacrimal gland can be evaluated by determination of level of tear lactoferrin using the same standards regardless of differences in pathogenesis of underlying diseases.

[A study of concentrations of CDDP in blood and cancer tissues in CDDP-treated gastrointestinal cancer].

Cisplatin (CDDP) was administered by intravenous drip in 3 cases of gastrointestinal cancer, and by intraperitoneal spraying in 2 other similar cases; and free CDDP and total CDDP levels in blood were determined with time. In the former cases, the free CDDP level rose from 30 minutes after the start of intravenous drip, to reach a peak at 2 approximately 4 hours after the end of the drip, while it was undetectable at 24 hours. The total CDDP level reached a peak 4 hours after the start of the drip, and then gradually decreased at 24, 48, 72 and 96 hours, although it still remained detectable in blood. In the latter cases, free CDDP appeared in blood at 15 minutes after the spraying, and disappeared from blood in 30 minutes to 1 hour. The total CDDP level reached a peak at 30 minutes, and then decreased in a manner similar to that seen in cases given the intravenous drip. CDDP levels in resected stomachs, resected colon and dissected lymph nodes following intravenous drip of CDDP were then measured in 4 cases of stomach cancer and another of cancer of the descending colon. There was no significant difference in CDDP level between cancer tissues, non-cancer tissues and lymph nodes. It thus appears that CDDP is taken up equally by normal tissues and cancer tissues, and that it may be necessary to study the metabolic pathways of CDDP after it has been taken up by the tissues.

c-myc Gene amplification in primary stomach cancer.

Fourteen human primary stomach cancer tissues were screened by Southern blot hybridization using six oncogene probes (myc, myb, H-ras, K-ras, abl, mos), and an amplification of c-myc oncogene was found in one tissue. This is the first report of c-onc amplification in primary stomach cancer tissue.