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Epidermolysis bullosa acquisita diagnosed by direct immunoelectron microscopy of the conjunctiva.

OBJECTIVE: To describe for the first time the direct immunoelectron microscopic pattern of immune deposits on the conjunctival basement membrane in epidermolysis bullosa acquisita (EBA). DESIGN: Case reports. PARTICIPANTS: Two patients. INTERVENTION: Epidermolysis bullosa acquisita associated with cicatrizing conjunctivitis. MAIN OUTCOME MEASURES: Direct immunofluorescence and direct immunoelectron microscopy without freezing on conjunctival and skin biopsy specimens, indirect immunofluorescence, Western immunoblot analysis. RESULTS: Results of direct immunoelectron microscopic examination of the conjunctiva showed the presence of immune deposits in the anchoring fibril zone, just beneath the lamina densa, in both patients. This finding was the same as the direct immunoelectron microscopic pattern shown in the skin of these patients, which is known to be very specific for EBA. Direct immunofluorescence was positive in the conjunctiva of only one patient. Indirect immunofluorescence and Western immunoblot analysis failed to detect circulating autoantibodies. CONCLUSIONS: Direct immunoelectron microscopy on the conjunctiva is a useful diagnostic tool to differentiate EBA from other related autoimmune mucocutaneous blistering diseases.

Adult↗

The prevalence of herpes family virus DNA in the conjunctiva of patients positive and negative for human immunodeficiency virus using the polymerase chain reaction.

OBJECTIVE: To help understand the pathogenesis of herpes family virus ocular infection among patients positive for HIV, the authors compared the rates of detection of herpes family virus DNA from the conjunctiva of patients who are positive and negative for human immunodeficiency virus (HIV) using the polymerase chain reaction (PCR). DESIGN: Cross-sectional study. PARTICIPANTS: The conjunctival scrapings of 30 patients positive for HIV and 30 patients negative for HIV were examined. INTERVENTION: PCR was used to assay for the presence of herpes simplex virus type 1 (HSV), varicella-zoster virus (VZV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV) DNA (n = 240 samples). MAIN OUTCOME MEASURE: The rate of detection of virus DNA in the two groups, controlling for age, gender, and race, was measured. RESULTS: HSV and VZV DNA were not detected in any of the HIV-positive or HIV-negative samples. CMV DNA was detected in 20% (6 of 30) of patients positive for HIV and was undetected in control subjects negative for HIV (P = 0.01). EBV DNA was detected in 40% (12 of 30) of patients positive for HIV and in 47% (14 of 30) of control subjects negative for HIV (P = 0.58). CONCLUSIONS: There was no difference in the frequency of detection of HSV, VZV, or EBV DNA from the conjunctiva of patients positive or negative for HIV. Only CMV DNA was detected at a significantly higher rate in the conjunctiva of patients positive for HIV compared with control subjects negative for HIV. These different rates of peripheral virus shedding may be one possible explanation for the different rates of clinical infection among the herpes family viruses among patients positive for HIV.

Adult↗

Arginine vasopressin transport and metabolism in the pigmented rabbit conjunctiva.

The purpose of this study was to evaluate the transepithelial transport and metabolism of arginine vasopressin (AVP) in the pigmented rabbit conjunctiva, both in the absence and presence of protease inhibitors. The apparent permeability coefficient, P(app), for 3H-AVP was determined in the modified Ussing chamber, and AVP metabolites were monitored by reversed phase HPLC using a C18 column. At 50 nM donor 3H-AVP, the P(app) in the mucosal-to-serosal (ms) direction was about five times higher than that in the opposite direction. Excess (0.1 mM) AVP decreased the P(app) for labelled AVP in the mucosal-to-serosal (ms) direction by about 50%. However, intact AVP transport showed neither concentration nor direction dependence. HPLC analysis revealed two subspecies of 3H-AVP in the receiver fluid and virtually no degradation products in the donor fluid following 3 h flux experiments. 3H-AVP transported in the ms direction underwent extensive hydrolysis (73%), which was decreased by 33% with mucosal application of 2 mM camostat mesylate (an aminopeptidase inhibitor) or by 27% with 0.5 mM leupeptin (a serine protease inhibitor). By contrast, 3H-AVP transported in the serosal-to-mucosal (sm) direction resulted in only 37% hydrolysis, and mucosal application of either inhibitor did not significantly affect the P(app) for intact AVP. These data suggest that intact AVP transport in the conjunctiva may be mediated mostly by passive diffusion and enzymatic degradation of AVP may be mediated by proteolytic enzymes present on the mucosal side of the conjunctiva.

Animals↗

The importance of cellular changes in the conjunctiva of patients with uveitic glaucoma undergoing trabeculectomy.

The results of trabeculectomy in patients with glaucoma secondary to uveitis are poor, but the exact cause of this remains unknown. Therefore, a study was undertaken to determine differences between conjunctiva from patients with glaucoma secondary to uveitis and that from patients with primary open angle glaucoma. Conjunctival biopsies from three patient groups were quantitatively analysed by light microscopy. Group A consisted of 10 patients with glaucoma secondary to uveitis who, prior to surgery, had been treated with topical steroids and topical beta-blocker drugs. Control groups (groups B and C) each consisted of 10 patients with primary open angle glaucoma for whom primary trabeculectomy (group B) or trabeculectomy following failure of topical beta-blockers alone (group C) was planned. Treatment with topical beta-blocker drugs was found to redistribute mast cells within the conjunctiva but to have no significant effect on cell numbers. In comparison with either control group, uveitic conjunctiva was found to contain significantly more fibroblasts, lymphocytes and macrophages. It is possible that these differences are responsible for an enhanced risk of excessive external bleb fibrosis, failure of filtration surgery and poor results with adjunctive 5-fluorouracil.

Adrenergic beta-Antagonists↗

The effect of topical cyclosporin on conjunctiva-associated lymphoid tissue (CALT).

Topical cyclosporin A is increasingly being used in the treatment of ocular surface immune-mediated disorders. The availability of the drug in oil-based vehicles or collagen shields has restricted its use because of ocular irritation or blurring of vision. Although topical cyclosporin is being used more frequently, its effect on the immunocompetent cells of the conjunctiva is not known. Our aim was to study the effect of cyclosporin instillation on the immunocomponent cells of conjunctiva-associated lymphoid tissue (CALT) of Lewis rat, using a novel method of topical drug delivery. A suspension of collagen bits impregnated with cyclosporin A was instilled into eyes of Lewis rats for 4 days (group 1) or 8 days (group 2). Control rats (group 3) received the suspension without cyclosporin. Frozen sections of eyelids and conjunctiva were immunostained with the following monoclonal antibody markers: W3/13 (CD3), W3/25 (CD4, macrophages), OX-8 (CD8), MARD-3 (B cells), ED1, ED2 (macro/monocytes), OX-6 (class II MHC, Ia) and OX-39 (CD25, IL-2 receptor). Intraepithelial (IE) and substantia propria cells for each subset were counted and expressed as numbers per section. By day 8, intraepithelial and substantia propria cells for all the above markers, except B cells, showed a significant reduction in numbers. The p values were < 0.02 for W3/13 (CD3), W3/25 (CD4), OX-8 (CD8), OX-39 (CD25) (IE only), ED1, ED2 and OX-6 positive cells. Goblet cells of control animals showed strong positive reaction with OX-39 (CD25) antibody. This was completely abolished following 8 days of topical cyclosporin. This study demonstrated that topical cyclosporin A induces a marked reduction in numbers of all subtypes of immunocompetent cells in the conjunctival epithelium and substantia propria.

Administration, Topical↗

Expression of growth factors in the conjunctiva from patients with active trachoma.

PURPOSE: The blinding complications of trachoma are associated with progressive conjunctival fibrosis due to excessive accumulation of extracellular matrix (ECM) components. We studied the processes involved in the regulation of fibrosis in trachoma by investigating the expression of the fibrogenic and angiogenic connective tissue growth factor (CTGF) and basic fibroblast growth factor (bFGF), the angiogenic vascular endothelial growth factor (VEGF), the angiogenesis-associated endothelial cell marker CD105 (endoglin), and the ECM protein tenascin in the conjunctiva. METHODS: Conjunctival biopsy specimens from six patients with active trachoma, and six control subjects were studied by immunohistochemical techniques using monoclonal and polyclonal antibodies directed against CTGF, bFGF, VEGF, CD105, and tenascin. RESULTS: In the normal conjunctiva, weak immunoreactivity for VEGF was observed in epithelial cells. There was no immunoreactivity for the other antibodies. In all trachoma specimens, immunoreactivity for CTGF and bFGF was localized in monocytes/macrophages, positive for the CD68 marker. Strong immunoreactivity for VEGF was observed in epithelial cells and on vascular endothelial cells. CD105 immunoreactivity was observed on vascular endothelial cells. Immunoreactivity for tenascin was noted in the upper substantia propria. CONCLUSIONS: These findings suggest that macrophages play an active role in conjunctival scarring, upregulated local production of CTGF, bFGF, and VEGF contributes to both fibrous tissue growth and angiogenesis, vascular endothelial cells are activated and are undergoing active angiogenesis, and deposition of tenascin reflect remodelling of the conjunctiva in trachomatous conjunctivitis.

Adolescent↗

Pterygium treatment using 5-FU as adjuvant treatment compared to conjunctiva autograft.

BACKGROUND: The use of conjunctiva autograft, adjunct antimetabolite therapy has been shown to be effective in preventing pterygium recurrence. OBJECTIVE: To compare 5 fluorouracil (5-FU) to conjunctival autograft in the treatment of large, fleshy pterygium. METHODS: A randomised controlled prospective study of outcome of pterygium treatment using 5-FU as adjuvant treatment compared to conjunctiva autograft. Thirty-five eyes with large pterygium treated with bare sclera conjunctival excision plus 5-FU were compared with 33 eyes treated with excision and conjunctival autograft alone. RESULTS: Post-operative pterygium recurrence was observed in four (11.4%) eyes treated with 5-FU and 4 (12.1%) eyes treated with conjunctiva autograft (P>0.05). The post-operative complications included, granuloma formation 11.4% for 5-FU and 3.0% for autograft and conjunctival discharge 5.7% for 5-FU group only. CONCLUSION: 5-FU is marginally superior to conjunctival autograft in the prevention of pterygium recurrence but neither gives a more desirable single digit recurrence rate. Randomised studies combining both conjunctival autograft and 5-FU in pterygium treatment is advocated to further explore their effect.

Adult↗

[General metastases in squamous cell carcinoma of the conjunctiva].

BACKGROUND: Squamous cell carcinoma of the conjunctiva is rare. Together with epithelial dysplasia of the conjunctiva the incidence is reported to be 1.9 per 100,000 population and year. After adequate therapy total cure is often achieved. Generalised metastases are however extremely rare. PATIENT AND METHOD: The case of a 71-year old white male patient is presented. Because of a recurrent conjunctival tumour in his right eye local resections had to be performed. The histopathological diagnosis was epithelial dysplasia. After a 3-year history a squamous cell carcinoma developed. In spite of multiple operations total excision of the tumour could not be achieved. Exenteration was refused by the patient. After radiotherapy of the orbit a temporary improvement was observed. However, 9 years later the patient died of general metastases to the long bones, the spine, and the ribs. RESULTS: Histological examination of material obtained during hemilaminectomy of the vertebrate bodies 8 and 9 confirmed the diagnosis of metastases of a squamous cell carcinoma. CONCLUSION: Although extremely rare, general metastases after squamous cell carcinoma of the conjunctiva must be kept in mind. This has to be considered when obtaining informed consent of the patient, planning adequate therapy and follow-up.

Aged↗

[Radical conjunctivectomy in therapy of malignant melanoma of the conjunctiva].

BACKGROUND: In tumor excision of large malignant melanomas of the conjunctiva or in the excision of widespread precancerous melanosis the development of symblepharon is a threatening complication after conjunctivectomy, especially if the conjunctival fornix is afflicted. PATIENTS: Three patients with suspected nodular melanoma and surrounding precancerous melanosis of the conjunctiva were treated by extended excision of the conjunctiva. Instead of a conjunctival graft, spontaneous epithelization under the protection of an 'Illig' contact lens was achieved. After this, two patients with histologically proven malignant melanomas were treated by beta-ray radiation with 90-Strontium. RESULTS: Within four weeks after the excision a complete epithelization of the wound occurred and the functional as well as the cosmetic results were very satisfying. During the follow-up between 12 and 22 months no recurrence of the melanomas and no formation of symblepharon was observed. CONCLUSIONS: With regard to these results extended tumor excision without conjunctival graft seems to be a reasonable therapy for large nodular melanomas and widespread melanosis, while the application of an 'Illig' contact lens avoids the formation of symblepharon.

Adult↗

Kinetic evidence for Na(+)-glucose co-transport in the pigmented rabbit conjunctiva.

PURPOSE: To obtain kinetic evidence for the existence of a Na(+)-coupled glucose co-transport process on the mucosal (tear) side of the pigmented rabbit conjunctiva. METHODS: The excised pigmented rabbit conjunctiva was mounted in the modified Ussing chamber for measurement of 22Na and 3H-3-O-methyl-D-glucose (3-O-MG) fluxes. RESULTS: In the presence of 5 mM glucose, the conjunctival tissue showed net Na+ absorption in the mucosal-to-serosal direction at an approximate rate of 0.15 microEq/cm2/h. This net Na+ absorption was abolished by serosally added 0.5 mM ouabain, but not affected by mucosally added 0.1 mM or 1 mM amiloride. There was a 40-60% reduction in net Na+ absorption under the glucose-free condition or in the mucosal presence of 0.5 mM phlorizin. Moreover, serosally added ouabain and mucosally added phlorizin (both at 0.5 mM) significantly decreased the 3-O-MG permeability coefficient in the mucosal-to-serosal direction by about 70%, whereas mucosally instilled 0.1-1.0 mM amiloride was without any effect. Three-O-MG absorption in the mucosal-to-serosal direction appeared to be coupled with Na+ transport with a 1:1 stoichiometry. In addition, this process exhibited temperature dependency, saturability, and directionality. CONCLUSION: Our findings are consistent with Na(+)-glucose cotransport as being one of the mechanisms for mucosal Na+ entry into the epithelial cells of the pigmented rabbit conjunctiva.

3-O-Methylglucose↗

Expression of nuclear retinoic acid receptor and retinoid X receptor mRNA in the cornea and conjunctiva.

PURPOSE: The effects of retinoic acid in cells are mediated by the nuclear retinoic acid receptors (RARs) and retinoid X receptors (RXRs). Although vitamin A is essential for the normal development and maintenance of the ocular surface, the RARs and RXRs have not been studied in cornea and conjunctiva. The purpose of this study was to identify the mRNA for these receptors in corneal and conjunctival cells in culture and to determine whether all-trans retinoic acid is able to induce expression of RAR mRNA. METHODS: Total RNA was extracted from cultured rabbit corneal stroma and conjunctival fibroblasts and rabbit corneal epithelial cells. RNA was analyzed by Northern blotting using the cDNA probes for RAR alpha, RAR beta, RAR gamma, RXR alpha, RXR beta and RXR gamma mRNA. To investigate induction of retinoid receptors, cells were exposed to 10(-6) M all-trans retinoic acid for 2-48 h before preparation of RNA. Effects of retinoic acid on cell proliferation were also investigated. RESULTS: RAR alpha mRNA transcripts (3.7 kb), RAR beta mRNA transcripts (3.3 kb) and RAR gamma mRNA transcripts (3.3 kb) are expressed by all the cell types studied, as are the RXR alpha mRNA transcripts (5.0 kb) and RXR beta mRNA transcripts (3.3 kb). RXR gamma mRNA is not detectable in corneal and conjunctival cells. All-trans retinoic acid induced RAR beta mRNA expression in corneal and conjunctival fibroblasts. Increased mRNA levels were detectable after 4-8 h and peaked by 24 h. RAR beta mRNA was not induced by retinoic acid in corneal epithelial cells. Retinoic acid also inhibited proliferation of conjunctival and corneal fibroblasts but had no effect on growth of corneal epithelial cells. CONCLUSIONS: The expression of RARs and RXRs in the cornea and conjunctiva is similar to that reported in other tissues. The identification of these receptors may lead to a better understanding of gene transcription pathways in the cornea and conjunctiva and of the mechanisms that control keratinization, differentiation and proliferation of the cells of these tissues. The data suggest a relationship between the induction of RAR beta mRNA expression and inhibition of cell proliferation by retinoic acid.

Animals↗

Predominance of infiltrating IL-4-producing T cells in conjunctiva of patients with allergic conjunctival disease.

PURPOSE: Previous reports have suggested that type 2 cytokine responses at the site of inflammation are important in the pathogenesis of allergic disease. In this study, we investigated the frequency of IFN-gamma- or IL-4-producing T cells from conjunctiva or peripheral blood mononuclear cells (PBMC) from patients with allergic conjunctival diseases. METHODS: We obtained conjunctival samples using Cytobrush and peripheral blood samples from the patients with allergic conjunctival disease. Conjunctival samples and PBMC were stimulated with phorbol myristate acetate and calcium ionophore. Frequencies of cytokine-producing T cells were analyzed by flow cytometry based on the intracellular cytokine staining. RESULTS: The frequency of IL-4-producing conjunctival T cells in patients with vernal keratoconjunctivitis (VKC) and atopic keratoconjunctivitis (AKC) were significantly higher than that in patients with allergic conjunctivitis (AC). The frequencies of IL-4-producing conjunctival CD4+ T cells of the patients with AC, AKC, or VKC were significantly higher than those of peripheral blood CD4+ T cells of the same disease. Further, increased frequency of P-selectin ligand expressing IL-4-producing T cells was observed in conjunctiva compared to that in PBMC of the patients with allergic conjunctival diseases. The frequencies of IL-4-producing peripheral blood T cells in the patients with ACD were significantly higher than that in normal controls. CONCLUSION: These results suggest that IL-4-producing T cells that have infiltrated into the conjunctiva play an important role in the immunopathogenesis of allergic conjunctival diseases, and P-selectin ligand is involved in the entry of T cells into allergic inflammatory site.

Adult↗

Alpha2-adrenergic receptors are present in normal human conjunctiva.

PURPOSE: Despite the passage of medications, including antiglaucoma drugs, through the ocular surface, and despite the increasing relevance of neurogenic inflammation in the ocular surface, the presence of some neuroreceptors in the conjunctiva has not been ascertained. This study describes the presence of alpha2-adrenergic receptors in normal human conjunctiva. METHODS: Immunofluorescence microscopy, electrophoresis, and Western blot analyses were done in human conjunctival biopsies and rat control tissues. Antibodies against alpha2-adrenergic receptor subtypes alpha2A, alpha2B, and alpha2C were used. RESULTS: Immunoreactivity for alpha2A- and alpha2B-, but not alpha2C-adrenergic receptors was evenly distributed in epithelial cells of human conjunctiva cryosections. Immunoreactive bands were detected for the three alpha2-adrenergic receptor subtypes: a major band of 48-50 kDa and fellow bands of 65-67 kDa. CONCLUSIONS: Normal human conjunctival epithelial cells express alpha2A-, alpha2B-, and alpha2C-adrenergic receptors. Further studies to determine the functional implications of these receptors in ocular surface homeostasis are warranted.

Aged↗

Applying secondary ion mass spectrometry to the analysis of elements in goblet cells of conjunctiva.

We investigated the location of elements in the goblet cells of rat conjunctiva by analyzing ion images produced by secondary ion mass spectrometry (SIMS) and comparing them with those produced by energy dispersive X-ray analyser (EDX). Conjunctivas of normal Spraque-Dawley rats were quenched in propane prechilled liquid nitrogen. Semi-thin sections were made with a cryo-ultramicrotome, freeze-dried, carbon-coated and observed under a light microscope, SIMS and scanning electron microscope (SEM). In the element analysis by SIMS, images of positive ions were examined with an O2+ primary ion source and images of negative ions with a Ga+ ion source. The same sections were observed and analysed with SEM-EDX. Morphological features and images of elements with SIMS and EDX were compared. Na, Mg, K, and Ca were detected as positive ions and OH, CN, P, S, and Cl as negative ions with SIMS, but C, N, O, Na, Mg, P, S, Cl, K, and Ca were detected with EDX. The spatial resolution of SIMS in element location was higher than that of EDX. Many elements were clearly located in the goblet cells on ion images by SIMS. Element ion images were demonstrated more densely in goblet cells than in other parts within conjunctiva and by SIMS compared to EDX. SIMS is a useful method for the detection of elements and their locations in ocular tissues and cells.

Animals↗

MUC7 expression in the human lacrimal gland and conjunctiva.

PURPOSE: Several mucins including MUC1, MUC2, MUC4, and MUC5AC have been identified at the ocular surface and in tears. The lacrimal gland, however, is not generally considered a source of ocular mucin. Because the lacrimal glands are similar to the salivary glands, we hypothesized that the lacrimal gland would express MUC7, a distinctive salivary mucin. We report the presence of MUC7 RNA and protein in normal human lacrimal glands as determined by reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and Western blot analysis. METHODS: RNA from lacrimal glands and conjunctivae was isolated and subjected to RT-PCR with primers specific for MUC7. The identity of the PCR products was confirmed by sequencing. In situ hybridization with PCR product-based riboprobes was used to locate MUC7 transcripts in the lacrimal gland. MUC7 protein was detected by Western blot analysis. RESULTS: Of six normal human lacrimal glands from which relatively intact mRNA could be extracted, four expressed MUC7. Hybridization with an antisense riboprobe for MUC7 indicates the presence of MUC7 transcripts in the cytoplasm of acinar cells. Western blot analysis confirms expression of the protein in the lacrimal gland. The presence of MUC1, MUC4, and MUC5B was also demonstrated by RT-PCR in lacrimal gland tissue. MUC7 transcripts and protein were also detected in normal human conjunctivae. CONCLUSIONS: The mucin profile of the lacrimal gland resembles that of the salivary gland. Both RNA and protein corresponding to MUC7 are present in the normal human lacrimal gland. Reverse transcription-polymerase chain reaction indicates that other transcripts of MUC1, MUC4, and MUC5B are present as well. Ocular MUC7 is also produced by the conjunctival mucosa. The lacrimal gland, therefore, contributes not only to the aqueous component of tears but also, in concert with the conjunctiva, may contribute to the total pool of ocular surface mucins.

Aged↗

Histamine receptors and the conjunctiva.

PURPOSE OF REVIEW: The purpose of this review is to evaluate the effect of histamine on various receptors in the conjunctiva. A Medline search from 1980 was performed on the histamine receptor subtypes H1, H2 and H3 in the human conjunctiva. RECENT FINDINGS: In the conjunctiva, histamine has been shown to induce various physiological and immunological changes through both H1 and H2 receptor stimulation. Histamine binding to conjunctival H1 receptors through the phospholipase C-dependent inositol phosphate pathway leads to the symptom of pruritus while histamine stimulation of the conjunctival H2 receptors has been indirectly shown to cause vasodilation. SUMMARY: The effect of histamine on conjunctival H1 receptors appears to be the primary target for ocular allergy treatment as it is primarily involved in ocular pruritus. The exact interaction of the conjunctival H2 receptors appears to work in a complementary fashion to the H1 receptor in controlling other features of ocular allergy such as vasodilation and injection. Thus, oral and topical antihistamines with multiple histamine receptor binding activities may provide an improved treatment paradigm for the various signs and symptoms of ocular allergy. The histamine H1, H2 and H3 receptor affinities of ketotifen, pyrilamine, and epinastine appear to have the strongest H1 and H2 affinities.

Animals↗

Heterogeneity in the ultrastructure of the mucous (goblet) cells of the rabbit palpebral conjunctiva.

AIM: The purpose of this study was to assess objectively the ultrastructure of the secretory granules in rabbit conjunctival mucin-producing 'goblet' cells. METHOD: The upper eyelids from five young adult dioestrous female rabbits were dissected out, stretched onto a cardboard support and prepared for transmission electron microscopy by repeated application of an isotonic two per cent glutaraldehyde fixative at room temperature. Post-fixation treatment included osmium tetroxide and staining with uranyl acetate and lead citrate. Low magnification micrographs were taken of the goblet cells of the conjunctiva, printed at a magnification of approximately 6,000 and the number, size and features of the secretory granules assessed. RESULTS: Across the entire palpebral conjunctiva of all five rabbits, the majority of mucous cells displayed a goblet shape and the secretory granules were uniformly pale in staining. The average width of the goblet cells was 10.8 +/- 1.1 microm and the diameter of the secretory granules was 0.82 +/- 0.16 microm. However, in localised regions across the palpebral conjunctiva of two of the rabbits, some goblet cells were different in that the secretory granules had either a denser-staining core, in which some of the granules were densely staining (while others were pale) or most of the granules were densely staining. These mucous cells had an average diameter of 10.3 +/- 1.7 microm and the granule diameters averaged 0.88 +/- 0.01 microm. For these abnormal goblet cells, inflammatory cells were found in their immediate vicinity. Occasionally, goblet cells were seen to be in the process of degranulation with associated apparent cell necrosis and the mucin granule diameter was close to 1 microm. CONCLUSIONS: The ultrastructure of the mucin-containing secretory granules of the conjunctival mucous cells is not necessarily homogeneous in character and further attention needs to be given to the effects of localised inflammation in the tissue and to possible hormonal influences.

Animals↗

Involvement of tachykinins in plasma extravasation induced by bradykinin and low pH medium in the guinea-pig conjunctiva.

1. The effect of bradykinin, capsaicin, substance P and low pH medium on plasma extravasation in the guinea-pig conjunctiva has been studied. Evans blue dye was measured in the conjunctiva after local instillation of the agents into the conjunctival sac. 2. Bradykinin (2-50 nmol), capsaicin (20-50 nmol) and substance P (0.5-5 nmol) caused a dose-dependent increase in plasma extravasation with the following order of potency: substance P > bradykinin = capsaicin. The effect of capsaicin (50 nmol) and substance P (5 nmol) was abolished by the tachykinin NK1 receptor antagonist, CP-99,994 (8 mumol kg-1, i.v.) (P < 0.01), whereas CP-100,263 (8 mumol kg-1, i.v.) the inactive enantiomer of CP-99,994 was without effect. CP-99,994 inhibited by 70% (P < 0.01) the effect of bradykinin. 3. The kinin B2 receptor antagonist, Hoe 140 (icatibant, 10 nmol kg-1, i.v.) abolished the response to bradykinin (50 nmol) (P < 0.01), but did not affect the responses to capsaicin (50 nmol) or substance P (5 nmol). Plasma extravasation induced by low pH medium (pH 1) was abolished by CP-99,994 (P < 0.01) and by Hoe 140 (P < 0.01). 4. The present findings suggest that: endogenous or exogenous tachykinins increase plasma extravasation in the guinea-pig conjunctiva by activation of NK1 receptors; bradykinin-induced plasma extravasation is mediated by tachykinin release from sensory nerve endings; low pH media cause plasma extravasation via release of kinins that by activation of B2 receptors release tachykinins from sensory nerve endings.

Animals↗