Human tear lipocalin acts as an oxidative-stress-induced scavenger of potentially harmful lipid peroxidation products in a cell culture system.
(73/947)
Human tear lipocalin [lipocalin 1 (lcn-1); von Ebner's gland protein] is a member of the lipocalin superfamily that is known to bind an unusual variety of lipophilic ligands. Because of its properties and its tissue-specific expression it has been suggested that lcn-1 might act as a physiological protection factor of epithelia. Overexpression of lcn-1 under certain disease conditions supported such a function. However, experimental investigations into its exact biological role and its mode of expression were impeded because lcn-1 was previously found to be produced only in serous glands. To overcome this problem we therefore sought a cell line that produced lcn-1 endogenously. Using reverse-transcriptase-mediated PCR analysis we found expression of lcn-1 in the human teratocarcinoma-derived NT2 precursor cells. Under normal conditions the production of lcn-1 is low. However, treatment of the cells with H(2)O(2) or FeSO(4), which typically induce lipid peroxidation, significantly enhanced the expression of lcn-1. Binding studies revealed that arachidonic acid and several lipid peroxidation products including 7beta-hydroxycholesterol, 8-isoprostane and 13-hydroxy-9,11-octadecadienoic acid specifically bind to lcn-1. To investigate the physiological consequence of this observation we purified holo-(lcn-1) from culture medium and extracted the bound ligands. The presence of F(2)-isoprostanes in the extracts obtained from the fractions containing lcn-1 indicates that these typical lipid peroxidation products are indeed ligands of the protein in vivo. These results support the idea that lcn-1 acts as a physiological scavenger of potentially harmful lipophilic molecules; lcn-1 might therefore be a novel member of the cellular defence against the deleterious effects of oxidative stress. (+info)
The effect of a new rigid gas-permeable contact lens design on lactic dehydrogenase activity in rabbit tears.
(74/947)
BACKGROUND: Most corneal damage induced by contact lenses is due to interference with corneal oxygenation. OBJECTIVE: To investigate the effect on the rabbit cornea of a rigid gas-permeable contact lens with a newly designed periphery. METHOD: We fitted New Zealand white rabbits (n = 12) with RGP contact lenses that were identical in all respects except for the design of the periphery. In each animal, one contact lens had an innovative periphery consisting of a microscopic diffractive relief lathed on the back surface; the other contact lens was of a conventional design. The lenses were worn continuously for 7 days. During this experimental period and for 1 additional week we assessed the corneal damage by daily testing lactic dehydrogenase activity in the tears. RESULTS: On the last day of the experimental week and the first 3 days of the healing period, mean tear LDH activity was significantly lower in the eyes with the new contact lens design than in eyes with the conventional lenses. CONCLUSIONS: The novel periphery design reduces corneal damage resulting from contact lens wear, as reflected by LDH levels in the tears. The new design probably facilitates the flow and exchange of tears under the contact lens, resulting in improved metabolism of the cornea. These findings may also prove applicable to soft contact lenses. (+info)
Evaluation of potentiating effect of a drop of lignocaine on tropicamide-induced mydriasis.
(75/947)
PURPOSE: To analyze whether preinstillation of lignocaine potentiates mydriasis by tropicamide in dark eyes and to determine possible mechanisms for this effect. METHODS: This investigation was conducted in two phases, the first being a double-masked, placebo-controlled, randomized clinical trial, enrolling 60 healthy dark brown eyes in 30 subjects aged 7 to 58 years. The control eye received a drop of (nonlignocaine) placebo before tropicamide 1%, and the contralateral study eye received a 4% lignocaine drop 3-minutes before the 1 drop of tropicamide was administered. A ruled pupillometer recorded pupil diameters every 10 minutes for 50 minutes. In phase II, to elucidate pathomechanisms after lignocaine, corneal and tear parameters were compared with baseline records in a further 60 such eyes. RESULTS: Pupillary diameters in the study eyes increased by 3.62 +/- 0.75 mm, significantly more than in the placebo (control) group (P = 0.000). Ninety percent of study eyes attained the clinically significant 6-mm size with preinstillation of lignocaine-many more than the 67% of control eyes (P = 0.016). The median time to achieve this critical 6-mm size was significantly faster in the study group (P = 0.005). In phase II, the 1 drop 4% lignocaine did not show corneal changes with slit lamp or fluorescein staining and did not reduce media clarity or induce a significant change in tear pH. It markedly decreased Schirmer values (P = 0.000), reduced tear break-up time (P = 0.003), and increased corneal thickness measured by optical pachymetry (P = 0.010). CONCLUSIONS: The phase II findings indicate corneal microepithelial damage and reduced tearing. Both may enhance intraocular penetration and hence potentiation of tropicamide. This remarkable phenomenon could find use with many other important topical medications. (+info)
Lacrimal immunoglobulins and complement quantified by counter-immunoelectrophoresis.
(76/947)
Immunoglobulin and complement concentrations of tears were measured using a sensitive electroimmunodiffusion assay. In the 13 subjects tested, the predominant immunoglobulin was secretory IgA (mean concentration 107 mug/mg protein; standard error of mean 15; n equal 17). The B1C component of complement was measurable in half of the subjects studied. The data illustrate that it is possible to quantify exactly certain humoral indices which may be important in the host defense of the eye. (+info)
Role of tears in keratocyte loss after epithelial removal in mouse cornea.
(77/947)
PURPOSE: To study the role of tears in the death of keratocytes after epithelium removal in the mouse cornea. METHODS: In anesthetized mice, an approximately 1-mm circle of epithelium was removed from the center of the cornea, exposing the underlying stroma. In one group of animals, access of tears to the bare stroma was allowed-in vivo, by closing the eyelids, or ex vivo, by dropping tears from another animal onto the denuded stroma of an enucleated eyeball. In another group, tear access was denied-in vivo, by bathing the cornea continuously in saline or by keeping the lids open, or ex vivo, by rinsing the denuded cornea before incubating the enucleated eyeball. In a separate group, corneal epithelial debris from another mouse was placed on the bare stroma of an enucleated eyeball. The corneas were isolated, stained with a fluorescent nuclear dye, and observed en face in a wholemount preparation under a fluorescence microscope, to evaluate the distribution of intact nuclei across the entire depth of the stroma. RESULTS: Between 1.5 and 2 hours after exposure to tears, the nuclei of the anterior keratocytes under the area of epithelial debridement invariably degenerated. When they had been protected from the tears, however, no degeneration was observed. Epithelial debris applied on the bare stroma had no effect on the underlying keratocytes. CONCLUSIONS: Factors in tear fluid trigger keratocyte loss after removal of the epithelium in the mouse cornea. (+info)
Immune response to chlamydial 60-kilodalton heat shock protein in tears from Nepali trachoma patients.
(78/947)
Although the host immune response to the 60-kDa chlamydial heat shock protein (hsp60) has been implicated in trachoma pathogenesis, no studies have examined mucosal immune responses to hsp60 in populations for which chlamydia is endemic. Tears and sera from Nepali villagers were reacted against hsp60 fusion proteins, whole hsp60, and the major outer membrane protein (MOMP). Tears from villagers without disease were anti-hsp60 immunoglobulin G (IgG) reactive in 6 (38%) of 16 villagers compared with 36 (90%) of 40 with follicular trachoma (TF) (P < 0.001); 47 (89%) of 53 with inflammatory trachoma (TI) (P < 0.001); and 31 (84%) of 37 with conjunctival scarring (TS) (P = 0.002). By multivariate analysis, odds ratios for tear hsp60 IgG immunoreactivity in villagers with TF, TI, and TS were 49.2 (confidence interval [CI], 2.7 to 898), 22.6 (CI, 3 to 170), and 13.6 (CI, 1.4 to 133), respectively. There were no significant differences for tear IgA or secretory IgA (sIgA) reactivity to hsp60 or for tear sIgA and IgG reactivity to MOMP. Serum anti-hsp60 IgG immunoreactivity was associated with TI only. These data suggest that anti-hsp60 IgG immunoreactivity represents largely locally derived antibodies, which may promote disease pathology. In contrast, nonspecific high rates of anti-hsp60 sIgA antibodies suggest chronic or repeat stimulation from an endemic source of organisms. (+info)
Focal sialadenitis in patients with ankylosing spondylitis and spondyloarthropathy: a comparison with patients with rheumatoid arthritis or mixed connective tissue disease.
(79/947)
OBJECTIVES: To investigate the occurrence of and risk factors for focal sialadenitis in patients with rheumatoid arthritis (RA), mixed connective tissue disease (MCTD), ankylosing spondylitis (AS), and spondyloarthropathy (SpA). METHODS: A total of 85 patients (25 with RA, 19 with MCTD, 19 with AS, 22 with SpA) participated in the study. Each patient filled out a questionnaire for eye and oral symptoms and for the use of medication, and was interviewed; other tests included Schirmer's test, laboratory tests, collection of unstimulated and stimulated whole saliva, and minor salivary gland biopsy. A focus score of > or =1 was regarded as an indicator of focal sialadenitis. RESULTS: Focal sialadenitis was observed in 68% (57/84) of all patients. It affected 80% (20/25) of the patients with RA, 94% (17/18) of those with MCTD, 58% (11/19) of those with AS, and 41% (9/22) of those with SpA (chi(2) test, p=0.0013). Salivary secretion correlated negatively with the focus scores-that is, severity of focal sialadenitis. Patients with focal sialadenitis had both decreased salivary secretion and decreased tear secretion significantly more often than did patients without (chi(2) test, p=0.0074 and p=0.048 respectively). Patients with positive rheumatoid factor (RF), antinuclear antibodies (ANA), or SSA or SSB antibodies had sialadenitis significantly more often than did patients with negative antibodies. In the subgroup of patients with AS or SpA, no associations were found between focal sialadenitis and the presence of these antibodies. CONCLUSION: In addition to patients with RA or MCTD, focal sialadenitis also affects a very high proportion of patients with AS or SpA. Focus scores are significantly higher in patients with RA or MCTD than in those with AS or SpA. A significant association exists between focal sialadenitis and RF, ANA, SSA and SSB. However, in the subgroup of patients with AS or SpA, no associations were found between focal sialadenitis and serological markers or clinical symptoms. (+info)
Detection of natural peptide antibiotics in human nasolacrimal ducts.
(80/947)
PURPOSE: To determine the expression and production of antimicrobial peptides by mucosal cells of the lacrimal passage in healthy and pathologic states. METHODS: Detection of bactericidal-permeability-increasing protein (BPI), heparin-binding protein (CAP37), human cationic antimicrobial protein (LL-37), human alpha-defensin 5 (HD5), human alpha-defensin 6 (HD6), human beta-defensin 1 (HBD-1), and human beta-defensin 2 (HBD-2) was performed by reverse transcription-polymerase chain reaction (RT-PCR). Intracellular deposition of lysozyme, lactoferrin, secretory phospholipase A(2), human neutrophil defensins (HNP-1, -2, and -3), human beta-defensin 1 (HBD-1), and human beta-defensin 2 (HBD-2) was analyzed immunohistochemically. Samples were obtained from 15 patients by surgery and from 10 cadavers. RESULTS: RT-PCR revealed BPI, CAP37, and HBD-1 mRNA in samples of healthy nasolacrimal duct epithelium. Additionally, HBD-2 mRNA was detected in epithelial samples from patients with dacryocystitis. Messenger RNAs for LL-37 and alpha-defensin 5 and 6 were absent in all samples investigated. Immunohistochemistry revealed lysozyme, lactoferrin, secretory phospholipase A(2), and HNP-1, -2, and -3 to be present in all samples, whereas HBD-1 was present only in some of the healthy and inflamed samples. Immunoreactive HBD-2 peptide was visible only in some of the inflamed samples. CONCLUSIONS: The data suggest that the human efferent tear ducts produce a broad spectrum of antimicrobial peptides. Under inflammatory conditions, changes in the expression pattern occurred, revealing induction of the human inducible defensin HBD-2 and in some cases downregulation of HBD-1 and CAP37. Antimicrobial peptides have a therapeutic potential in dacryocystitis, in that they have a broad spectrum of antimicrobial activity and accelerate epithelial healing. However, caution is appropriate, because defensins also promote fibrin formation and cell proliferation, which are key elements in scarring processes, such as dacryostenosis. (+info)