muramidase and Dry-Eye-Syndromes

To compare the role of topically applied serum therapy with preservative-free artificial tear (AT) drops in patients with moderate to severe dry eye in Hansen's disease along with change in tear protein profile.. 144 consecutive patients were randomly divided into three groups. After a baseline examination of clinical parameters, each of the patients received designated modality of topical therapy six times a day for 6 weeks. Post-treatment documentation of clinical parameters was done at 6 weeks, and then at 12 weeks after discontinuation of topical therapy. Analysis of three tear proteins using gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis) was done at baseline, at the first and second post-treatment visits.. In the cord blood serum (CBS) group, except for McMonnies score and staining score, all other clinical parameters showed continued improvement in the first and second post-treatment analyses. In the autologous serum (ALS) group, all the clinical parameters except Schirmer's I showed significant improvement in the first post-treatment analysis .This was sustained at a significant level in the second analysis except for tear film break-up time (TBUT) and conjunctival impression cytology grading. In the AT group, all the parameters improved at a non-significant level except for TBUT in the first analysis. In the next analysis, apart from McMonnies score and TBUT, other clinical parameters did not improve. In the ALS and CBS groups, tear lysozyme, lactoferrin levels improved in both post-treatment measurements (statistically insignificant).Total tear protein continued to increase at statistically significant levels in the first and second post-treatment analyses in the CBS group and at a statistically insignificant level in the ALS group. In the AT group, the three tear proteins continued to decrease in both the analyses.. In moderate to severe dry eye in Hansen's disease, serum therapy in comparison with AT drops, improves clinical parameters and causes betterment in tear protein profile.. CTRI/2013/07/003802.

Topics: Administration, Topical; Adult; Aged; Conjunctiva; Double-Blind Method; Dry Eye Syndromes; Electrophoresis, Polyacrylamide Gel; Eye Infections, Bacterial; Eye Proteins; Female; Fetal Blood; Humans; Lactoferrin; Leprosy; Male; Middle Aged; Muramidase; Prospective Studies; Proteomics; Serum; Tears

The purpose of this study was to investigate the impact of using a rewetting drop (RWD) containing surface active agents (OPTI-FREE RepleniSH; Alcon, Fort Worth, TX) on the clinical performance and protein deposition when using a continuous-wear (CW) silicone hydrogel (SH) contact lens.. Subjects wore lotrafilcon A SH lenses on a 30-day CW basis for two consecutive 1-month periods while inserting either 0.9% unpreserved unit-dose saline (control) or multidose OPTI-FREE RepleniSH (test RWD). Subjective comfort and symptoms were assessed after 2 and 4 weeks with each product. After 1 month of wear with each product, lenses were collected and analyzed in the laboratory for total protein, total lysozyme, and percentage of denatured lysozyme.. Symptoms of dryness and comfort varied across the day regardless of drop type (p < 0.001) with dryness being maximal on waking, least in the middle of the day, and increased towards the evening. The test RWD provided greater comfort on insertion (p = 0.02), better visual quality (p < 0.01), and less mucous discharge on waking (p = 0.02) than the control product. Lysozyme deposition was significantly reduced after the use of the test RWD as compared to saline (0.73 +/- 0.5 microg/lens vs. 1.14 +/- 0.7 microg/lens; p < 0.001) as was total protein deposition (1.17 +/- 0.7 microg/lens vs. 1.86 +/- 0.8 microg/lens; p < 0.001). Lysozyme denaturation was also reduced with the use of the test RWD compared with the control (76 +/- 10% vs. 85 +/- 7%; p < 0.01).. The use of a RWD containing surface active agents provided greater subjective satisfaction, reduced lysozyme and total protein deposition, and reduced denatured lysozyme than a RWD containing saline alone.

Topics: Adult; Blotting, Western; Contact Lenses, Extended-Wear; Cross-Over Studies; Dry Eye Syndromes; Electrophoresis, Polyacrylamide Gel; Follow-Up Studies; Humans; Hydrogels; Muramidase; Ophthalmic Solutions; Prospective Studies; Prosthesis Design; Refractive Errors; Silicones; Surface-Active Agents; Tears; Treatment Outcome

To assess the efficacy of topical cyclosporine 0.05% (Restasis) in patients with dry eye associated with graft versus host disease after stem cell transplantation.. After completing a 3-month run-in period of using only artificial tears to control dry eye symptoms in both eyes, patients who failed to achieve adequate relief (n = 8) were instructed to instill topical cyclosporine twice a day. Visual acuity, slit-lamp appearance, and intraocular pressure were evaluated every 2 weeks for a minimum of 3 months. In addition, Schirmer basal secretion tests, noninvasive fluorescein breakup time, and tear lysozyme were also performed. Patients were also given a dry eye questionnaire regarding symptoms of burning, tearing, and blurred vision.. Dry eye signs improved significantly with cyclosporine treatment in 7 of 8 patients. Cyclosporine provided statistically significant improvements in Schirmer basal secretion scores (P = 0.003), tear breakup time (P = 0.002), and tear lysozyme levels (P = 0.033) after 3 months of treatment.. The findings in this prospective study suggest that dry eye associated with graft versus host disease can be effectively treated with topical cyclosporine, especially in patients unresponsive to other treatment modalities. These findings should be further evaluated in large-scale, controlled clinical trials.

Topics: Administration, Topical; Adult; Cyclosporine; Dry Eye Syndromes; Female; Graft vs Host Disease; Humans; Immunosuppressive Agents; Male; Middle Aged; Muramidase; Prospective Studies; Surveys and Questionnaires; Tears; Treatment Outcome; Visual Acuity

Dry eye disease (DED) is a chronic condition characterized by ocular dryness and inflammation. The tear film lipid layer (TFLL) is the outermost layer composed of lipids and proteins that protect the ocular surface. However, environmental contaminants can disrupt its structure, potentially leading to DED. Although the importance of tear proteins in the TFLL functionality has been clinically recognized, the molecular mechanisms underlying TFLL-protein interactions remain unclear. In this study, we investigated tear protein-lipid interactions and analyzed their role in the TFLL functionality. The results show that lysozyme (LYZ) increases the stability of the TFLL by reducing its surface tension and increasing its surface pressure, resulting in increased TFLL evaporation and bacterial invasion resistance, with improved wettability and lubrication performance. These findings highlight the critical role of LYZ in maintaining ocular health and provide potential avenues for investigating novel approaches to DED treatment and patient well-being.

Topics: Dry Eye Syndromes; Humans; Lipids; Muramidase; Physical Phenomena; Tears

Topics: Actins; Adult; Antigens, CD20; Biomarkers; Conjunctival Diseases; Dry Eye Syndromes; Humans; Lacrimal Apparatus Diseases; Lactoferrin; Muramidase; Prospective Studies; S100 Proteins; Stevens-Johnson Syndrome

To detect the immunohistochemical changes in the main lacrimal glands of patients suffering from chronic ocular sequelae of Stevens-Johnson syndrome (SJS).. Histological sections of biopsies from the lacrimal gland of three chronic SJS patients (mean age, 33 years; 2 males) with severe dry eye disease (Schirmer = 0 mm) were assessed using double immunofluorescence techniques. Changes in the expression of secretory proteins lactoferrin (Lf), lysozyme (Ly), aquaporin 5 (AQP5), S-100, and early apoptosis marker (Annexin V) were studied.. Different morphological expressions of secretory proteins were present in the three samples. One sample had maintained the immunoreactivity for Lf, Ly, S-100, similar to healthy controls. Two samples had significantly reduced immunoreactivity for anti-Lf, anti-Ly, and S-100, the weakest being in the sample with distorted lobular architecture and mild interlobular fibrosis. AQP5 had a distinct vesicular intracytoplasmic immunoreactivity suggesting defective trafficking and integration of the protein to the apical membrane. There was no S-100 immunostaining in the acinar or ductal epithelium, whereas interstitial nerve fibers scattered in the periacinar region showed reduced immunoreactivity for S-100. There was strong Annexin V immunoreactivity in the nuclei of epithelial cells in the majority of acinar and ductal epithelia of all the samples, with distorted nuclear morphology in one sample.. Defective trafficking of AQP5 and variable expression of Ly, Lf, S-100 are the notable findings in the lacrimal glands of chronic SJS patients along with signs of early apoptosis. It suggests that the palpebral lobe of the lacrimal gland is involved in the pathological processes occurring in the conjunctiva of SJS patients.

Topics: Adult; Annexin A5; Apoptosis; Aquaporin 5; Biomarkers; Chronic Disease; Dry Eye Syndromes; Eye Proteins; Female; Fluorescent Antibody Technique, Indirect; Humans; Lacrimal Apparatus; Lactoferrin; Male; Muramidase; S100 Proteins; Stevens-Johnson Syndrome; Young Adult

To evaluate the concentration of tear lysozyme in individuals with Sjogren´s syndrome, meibomian gland dysfunction, and non-dry-eye disease.. Ninety subjects were recruited for this study, including 30 with Sjogren´s syndrome, 30 with meibomian gland dysfunction, and 30 with non-dry-eye disease. All subjects were referred to participate in the study based on a "dry eye" investigation. They underwent a complete ocular surface ophthalmic examination encompassing ocular surface disease index, biomicroscopy, tear break-up time, Schirmer test type I, conjunctival vital staining with fluorescein and lissamine green, tear lysozyme concentration, and impression cytology.. Clinical tests yielded the following results: ocular surface disease index Sjogren´s syndrome: 64.5 ± 22.6 meibomian gland dysfunction: 43.5 ± 21.4, non-dry-eye disease: 6.7 ± 4.3 (p=0.02 between groups); Schirmer I test (mm/5 min): Sjogren´s syndrome: 4.95 ± 2.25, meibomian gland dysfunction: 13.28 ± 1.53, non-dry-eye disease 13.70 ± 1.39 (p<0.01 Sjogren´s syndrome vs. non-dry-eye disease and p<0.01 meibomian gland dysfunction vs. non-dry-eye disease); tear break-up time (seconds): Sjogren´s syndrome: 3.97 ± 1.47, meibomian gland dysfunction: 3.95 ± 0.86, non-dry-eye disease: 7.25 ± 1.90 (p<0.01 Sjogren´s syndrome vs. non-dry-eye disease and p<0.01 meibomian gland dysfunction vs. non-dry-eye disease); Lissamine green score: Sjogren´s syndrome-dry-eye: 6.18 ± 2.14, meibomian gland dysfunction-dry-eye: 5.27 ± 1.27, non-dry-eye disease: 1.52 ± 0.97 (p<0.01 Sjogren´s syndrome vs. non-dry-eye disease and p<0.01 meibomian gland dysfunction vs. non-dry-eye disease); impression cytology score: Sjogren´s syndrome: 1.88 ± 0.92, meibomian gland dysfunction: 1.67 ± 0.56, non-dry-eye: 0.45 ± 0.44 (p<0.01 Sjogren´s syndrome vs. non-dry-eye disease and p<0.01 meibomian gland dysfunction vs. non-dry-eye disease) and; tear lysozyme concentration (µg/mL): Sjogren´s syndrome: 751.25 ± 244.73, meibomian gland dysfunction: 1423.67 ± 182.75, non-dry-eye disease: 1409.90 ± 188.21 (p<0.01 Sjogren´s syndrome vs. non-dry-eye disease and p<0.01 Sjogren´s syndrome vs. meibomian gland dysfunction).. The concentration of lysozyme in the tears was lower in Sjögren's syndrome patients than in meibomian gland dysfunction and non-dry-eye disease groups. Hence, the lacrimal lysozyme could be considered as a simple, non-invasive, and economical biomarker to differentiate between Sjögren's syndrome dry eye disease and meibomian gland dysfunction dry eye disease.

Topics: Dry Eye Syndromes; Humans; Meibomian Gland Dysfunction; Meibomian Glands; Muramidase; Sjogren's Syndrome; Tears

We report a method for sensing analytes in tear-fluid using commercial contact lenses (CLs) as sample collectors for subsequent analysis with a cost-effective and field-portable reader. In this study we quantify lysozyme, the most prevalent protein in tear fluid, non-specifically bound to CLs worn by human participants. Our mobile reader uses time-lapse imaging to capture an increasing fluorescent signal in a standard well-plate, the rate-of-change of which is used to indirectly infer lysozyme concentration through the use of a standard curve. We empirically determined the best-suited CL material for our sampling procedure and assay, and subsequently monitored the lysozyme levels of nine healthy human participants over a two-week period. Of these participants who were regular CL wearers (6 out of 9), we observed an increase in lysozyme levels from 6.89 ± 2.02 μg mL

Topics: Antiviral Agents; Contact Lenses, Hydrophilic; Dry Eye Syndromes; Humans; Muramidase; Tears

To evaluate the clinical efficacy of 3% diquafosol sodium ophthalmic solution for dry eye, and to analyze the concentration of tear proteins and mucin-like substances after the treatment.. Fifty eyes of 25 patients with dry eye syndrome were prospectively enrolled. The patients were treated with diquafosol solution at a dose of 1 drop in each eye 6 times daily for 4 weeks. The parameters of clinical efficacy were tear osmolarity, tear breakup time (BUT), fluorescein staining scores for the cornea and conjunctiva, Schirmer test values, and subjective symptoms evaluated using the ocular surface disease index (OSDI). Tears collected with Schirmer test strips were analyzed by high-performance liquid chromatography, and the concentrations of the total protein and the 4 major tear proteins, namely, secretory IgA, lactoferrin, lipocalin-1, lysozyme, and N-acetyl-neuraminic acid (Neu5Ac), were measured. Neu5Ac is a major sialic acid, a marker of secretory mucins.. The BUT, keratoconjunctival staining scores, and Schirmer test values were improved with statistical significance after the treatment with diquafosol solution, while changes in the other parameters, including tear osmolarity, corneal staining scores, and OSDI scores were not significant. The Neu5Ac concentration was significantly increased, which was not accompanied by changes in tear proteins.. Topical application of diquafosol significantly improved the clinical parameters of the BUT, keratoconjunctival staining scores, and Schirmer test values and was accompanied by increased sialic acid content in the tears of patients with dry eye.

Topics: Adult; Aged; Aged, 80 and over; Chromatography, High Pressure Liquid; Dry Eye Syndromes; Eye Proteins; Female; Humans; Immunoglobulin A, Secretory; Lactoferrin; Lipocalin 1; Middle Aged; Muramidase; N-Acetylneuraminic Acid; Ophthalmic Solutions; Osmolar Concentration; Polyphosphates; Prospective Studies; Purinergic P2Y Receptor Agonists; Tears; Uracil Nucleotides; Young Adult

Among the most frequently encountered pathologies examined by the ophthalmologist is dry eye syndrome (DE), which can be discovered particularly in the elderly. The initial diagnosis of DE is of high importance, but also challenging. This is because the biochemical changes in the tear film often develop before any detectable signs.. In this study, the possible relationship between ocular symptomatology, tear volume and tear break-up time (TBUT) and lipocalin, lactoferrin and lysozyme concentrations in the tear film were explored in a group of symptomatic dry-eyed postmenopausal (PM) women compared to age-matched controls.. Sixty-six healthy PM females with ages of at least 50 years were grouped in two homogeneous lots (by age, post-menopause, co-morbidities) of 33 females each, one lot presenting mild or moderate dry eye syndrome (DE) and one asymptomatic non-dry eye (NDE), based on their feedback to the Ocular Surface Disease Index (OSDI) questionnaire and noninvasive TBUT and Schirmer test results. Tears were collected via capillary tubes and an eye wash method. Tear lysozyme, lactoferrin and lipocalin concentrations were determined via electrophoresis.. OSDI responses revealed 3 mild DE, 30 moderate DE and 33 NDE. The OSDI total score and sub scores for the DE group were significantly greater than for the NDE group (p < 0.001). The mild and moderate DE group exhibited significantly shorter TBUTs compared to NDE (p < 0.001). No difference in tear lysozyme or lipocalin concentrations was found between DE and NDE groups, irrespective of the tear collection method, but a significant difference was found in lactoferrin concentration (p<0.001). No significant correlations were found between symptoms or signs of DE compared to either lipocalin, lysozyme or lactoferrin concentrations.. In a PM population, lipocalin and lysozyme are invariable, irrespective of the presence and severity of DE symptoms. However, lactoferrin shows a significant decrease. This is a comprehensive study of lipocalin, lactoferrin and lysozyme in dry-eyed PM women and our results suggested that lactoferrin could be used as a biomarker of DE in postmenopausal women.. PM = postmenopausal; DE = dry eye disease; NDE = non-dry eye; ELISA = Enzyme-linked immunosorbent assay.

Topics: Dry Eye Syndromes; Female; Humans; Lactoferrin; Lipocalin 1; Middle Aged; Muramidase; Postmenopause; Tears

This study investigates the extent of the human transcriptome that can be quantified from conjunctival impression cytology extracts. The aim is to determine if sufficient RNA can be isolated from a patient's conjunctival surface to identify differences in gene expression between dry eye and normal patients of (a) an array of 96 inflammatory biomarkers and associated receptors, and (b) if this comparison can be expanded to the entire transcriptome.. CIC was used to collect conjunctival surface cells from 53 qualifying normal and dry eye patients. Based on prior optimization of all assay steps, RNA was isolated from the samples using a Qiagen RNeasy Plus Mini Kit and qRT-PCR was used to determine gene expression of 96 genes using TaqMan Low Density Array cards. Samples from six normal and six dry eye patients were then assayed on an Illumina Human HT-12 BeadChip.. Optimization steps yielded an RNA processing procedure that improved yield from an initial 12 genes through 96, then to the entire human transcriptome. For the HT-12 BeadChip, more than 30 genes differed by a factor of >1.5 between the dry eye and normal groups and seven genes were down-regulated by a factor of >2.0 in the dry eye group: HLA-DRB5, PSCA, FOS, lysozyme, TSC22D1, CAPN13 and CXCL6.. Conjunctival impression cytology can be used to collect sufficient RNA from conjunctival surface cells that, when processed optimally, allows successful transcriptome-wide expression analysis. While the current transcriptome analysis used a limited patient group, larger studies of patients with various types and severities of dry eye should reveal significant gene expression trends that can then be targeted to improve dry eye treatment options.

Topics: Antigens, Neoplasm; Biomarkers; Calpain; Chemokine CXCL6; Conjunctiva; Dry Eye Syndromes; Female; Genome-Wide Association Study; GPI-Linked Proteins; HLA-DRB5 Chains; Humans; Male; Middle Aged; Muramidase; Neoplasm Proteins; Proto-Oncogene Proteins c-fos; Repressor Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA; Transcriptome

The aim of this study was to explore the possible interactions between vitelline membrane outer layer 1 homolog (VMO1) and other tear proteins and to determine the function of VMO1 in tear fluid.. Interactions between recombinant human VMO1 and several abundant tear proteins were determined by dot blot, His pull-down, immunoprecipitation, and Western blot assays, as well as by computer-assisted prediction and modeling of molecular interactions. Kirby-Bauer antibiotic testing was performed to determine whether VMO1 possesses antimicrobial activity. Tear samples were collected from dry eye patients and from healthy controls. The role of VMO1 in maintaining the stability of tear film was investigated by measurement of contact angles on Teflon, tear break-up time (TBUT) and the time-dependent reduction in tear film integrity in mice.. Vitelline membrane outer layer 1 homolog showed an interaction with lysozyme C (LYSC) in the dot-blot, His pull-down, and immunoprecipitation assays. Vitelline membrane outer layer 1 homolog revealed no zones of growth inhibition of standard strains of Staphylococcus aureus and Escherichia coli. Tears presented smaller contact angles on Teflon surfaces after the addition of VMO1 (P<0.05). Vitelline membrane outer layer 1 homolog-treated mice showed longer TBUTs (P<0.05). Tear films from VMO1-treated mice maintained their integrity for longer periods of time than tear films from the control group, and this effect was dose-dependent.. Vitelline membrane outer layer 1 homolog interacts with LYSC and has positive effects on the stabilization of tear film.

Topics: Animals; Blotting, Western; Disease Models, Animal; Dry Eye Syndromes; Egg Proteins; Humans; Immunoprecipitation; Mice; Mice, Inbred C57BL; Muramidase; Tears

Protein-meibum and terpenoids-meibum lipid interactions could be important in the etiology of meibomian gland dysfunction (MGD) and dry eye symptoms. In the current model studies, attenuated total reflectance (ATR) infrared (IR) spectroscopy was used to determine if the terpenoid β-carotene and the major proteins in tears and meibum affect the hydrocarbon chain conformation and carbonyl environment of wax, an abundant component of meibum. The main finding of these studies is that mucin binding to wax disordered slightly the conformation of the hydrocarbon chains of wax and caused the wax carbonyls to become hydrogen bonded or experience a more hydrophilic environment. Lysozyme and lactoglobulin, two proteins shown to bind to monolayers of meibum, did not have such an effect. Keratin and β-carotene did not affect the fluidity (viscosity) or environment of the carbonyl moieties of wax. Based on these results, tetraterpenoids are not likely to influence the structure of meibum in the meibomian glands. In addition, these findings suggest that it is unlikely that keratin blocks meibomian glands by causing the meibum to become more viscous. Among the tear fluid proteins studied, mucin is the most likely to influence the conformation and carbonyl environment of meibum at the tear film surface.

Topics: beta Carotene; Dry Eye Syndromes; Eye Proteins; Eyelid Diseases; Fatty Acids, Unsaturated; Humans; Keratins; Lactoglobulins; Meibomian Glands; Mucins; Muramidase; Protein Conformation; Spectroscopy, Fourier Transform Infrared; Viscosity

To evaluate changes in symptoms, objective tests, and signs after medical treatment of subjects with evaporative-type dry eye disease (EDE) caused by Meibomian gland dysfunction (MGD), and to analyze correlations among symptoms, signs and test results in the worse eyes (W-eyes) of the subjects.. Prospective clinical study of 21 symptomatic subjects with EDE caused by MGD. Subjects who were diagnosed with EDE in a first visit were treated for 6 weeks and re-evaluated in a second visit. The differences between initial and second visits were evaluated. Correlations among clinical symptoms, signs, and test results were performed using the data of the W-eyes. Variables evaluated included: dry eye symptoms, best corrected visual acuity (BCVA), contrast sensitivity, conjunctival hyperemia, phenol red thread test, tear break-up time (TBUT), tear meniscus height (TMH), corneal fluorescein and conjunctival rose Bengal staining, tear lysozyme concentration, Schirmer test, and lid margin assessment.. All items evaluated improved after treatment, but only conjunctival hyperemia and TMH improved significantly. TBUT and lid margin changes improved, but still remained abnormal. There were significant correlations among symptoms questionnaires and some clinical tests (TBUT, conjunctival hyperemia, TMH, and conjunctival rose Bengal staining).. Despite the instability of the tear film and lid margin alterations that continued after treatment, subjects with MGD improved symptomatically. The low degree of correlations among W-eye signs, symptoms, and tests reflects the independency of symptoms and signs in this complex pathology.

Topics: Adult; Aged; Blepharitis; Contrast Sensitivity; Cross-Sectional Studies; Dry Eye Syndromes; Female; Fluorescent Dyes; Humans; Hyperemia; Male; Meibomian Glands; Middle Aged; Muramidase; Ophthalmic Solutions; Prospective Studies; Rose Bengal; Surveys and Questionnaires; Tears; Visual Acuity

To analyze tear protein variations in patients suffering from dry eye symptoms in the presence of tear film instability but without epithelial defects.. Five microlitres of non-stimulated tears from 60 patients, suffering from evaporative dry eye (EDE) with a break-up time (BUT) <10 s, and from 30 healthy subjects as control (no symptoms, BUT >10 s) were collected. Tear proteins were separated by mono and bi-dimensional SDS-PAGE electrophoresis and characterized by immunoblotting and enzymatic digestion. Digested peptides were analyzed by liquid chromatography coupled to electrospray ionization quadrupole-time of flight mass spectrometry followed by comparative data analysis into Swiss-Prot human protein database using Mascot. Statistical analysis were performed by applying a t-test for independent data and a Mann-Whitney test for unpaired data (P<0.05).. In EDE patients vscontrols, a significant decrease in levels of lactoferrin (data in %+/-SD): 20.15+/-2.64 vs 24.56+/-3.46 (P=0.001), lipocalin-1: 14.98+/-2.70 vs 17.73+/-2.96 (P=0.0001), and lipophilin A-C: 2.89+/-1.06 vs 3.63+/-1.37 (P=0.006) was revealed, while a significant increase was observed for serum albumin: 9.45+/-1.87 vs 3.46+/-1.87 (P=0.0001). No changes for lysozyme and zinc alpha-2 glycoprotein (P=0.07 and 0.7, respectively) were shown. Proteomic analysis showed a downregulation of lipophilin A and C and lipocalin-1 in patients, which is suggested to be associated with post-translational modifications.. Data show that tear protein changes anticipate the onset of more extensive clinical signs in early stage dry eye disease.

Topics: Adipokines; Adult; Aged; Albumins; Carrier Proteins; Case-Control Studies; Dry Eye Syndromes; Electrophoresis, Polyacrylamide Gel; Eye Proteins; Female; Glycoproteins; Humans; Lactoferrin; Lipocalins; Male; Middle Aged; Muramidase; Myelin Proteins; Proteolipids; Proteomics; Secretoglobins; Tears; Uteroglobin

To investigate the potential relationship between subjective symptomatology, tear volume, and tear break up time with tear film lipocalin and lysozyme concentrations in a group of symptomatic dry-eyed postmenopausal (PM) women compared to age-matched controls.. Eighty-five healthy PM females (>50 years of age) were categorized as mild or moderate dry eye (DE), or asymptomatic [non-dry eye (NDE)] based on their responses to the Ocular Surface Disease Index (OSDI) questionnaire. Non invasive tear breakup time (NITBUT) and tear secretion were measured. Tears were collected via capillary tube and an eye wash method. Tear lysozyme and lipocalin concentrations were determined via Western blotting.. Ocular Surface Disease Index responses revealed 16 mild DE, 30 moderate DE, and 39 NDE. The OSDI total score and sub scores for the DE groups were significantly greater than for the NDE group (p < 0.001). The mild and moderate DE groups exhibited significantly shorter NITBUTs compared to NDE (p < 0.004). Tear secretion using the Phenol Red Thread (PRT) test was found to be significantly lower in the moderate DE group compared to NDE (p < 0.001). No difference in tear lysozyme or lipocalin concentration was found between DE and NDE groups, irrespective of tear collection method, although method of collection significantly influenced absolute concentrations (p < 0.008). Significant correlations were not found between symptoms or signs of DE compared to either lipocalin or lysozyme concentration.. Within a PM population, lipocalin and lysozyme are invariant, irrespective of the presence and severity of DE symptoms. This is the first comprehensive study of lipocalin and lysozyme in dry-eyed PM women and our results suggest that neither protein would offer utility as a biomarker of DE.

Topics: Aged; Blotting, Western; Dry Eye Syndromes; Eye Proteins; Female; Humans; Lipocalins; Middle Aged; Muramidase; Postmenopause; Severity of Illness Index; Surveys and Questionnaires; Tears

Adverse environmental conditions elicit dry eye (DE)-related signs and symptoms. The purpose of this work is to determine whether these conditions can alter a normal-to-borderline ocular surface in subjects with DE symptoms.. Ten minimally symptomatic contact lens (CL)-wearing subjects were exposed, without (WO-) and with (W-)CLs, to a controlled adverse environment (CAE) of 22.0 +/- 2.0 degrees C and 19.0% +/- 4.0% relative humidity (RH) for 2 hours in an environmental chamber (EC). One month later, the same subjects were placed in an indoor normal environment (INE) of 24.2 +/- 1.3 degrees C and 34.8% +/- 2.9% RH for 2 hours. DE-related signs and symptoms were evaluated before and after each exposure. The reversibility of changes provoked by CAE or INE was also evaluated.. Without CL wear, significant changes were found in DE signs (noninvasive tear break-up time [NIBUT], conjunctival hyperemia and phenol red thread test) after CAE exposure, but not found after INE exposure. However with CL wear, the same tests were altered after both CAE and INE exposure. Most of these changes returned to normal values within 1 month after environmental exposure.. Significant changes in comfort and the ocular surface tests were found after 2 hours of exposure to CAE. These results show the negative impact that an adverse environment, especially low RH, can have on the ocular surface. These alterations were fully reversible. This indicates that the CAE is a safe and functional condition in which to standardize DE diagnostic tests and evaluate therapeutics.

Topics: Adult; Atmosphere Exposure Chambers; Conjunctiva; Contact Lenses; Dry Eye Syndromes; Environment, Controlled; Environmental Exposure; Female; Hot Temperature; Humans; Humidity; Hyperemia; Male; Muramidase; Orbit; Tears

To determine whether intolerance to contact lens wear is attributable to clinical or protein characteristics of the tear film.. Thirty-eight subjects participated; 20 were successful contact lens wearers and 18 had discontinued contact lens wear because of discomfort. Baseline tear film (no lens wear) was analyzed with a range of clinical measurements and protein analyses (lactoferrin, sIgA, and lysozyme). Comfort was determined after 6 hours of lens wear, and differences in tear film characteristics between subject groups were determined. In half of the subject group (n = 19), discriminant analysis was used to develop an equation for predicting the likelihood of intolerance to lens wear. Sensitivity and specificity were determined by testing the formula on the remaining subjects. These formulas were also tested on a separate group of subjects enrolled in a contact lens-wearing trial.. Tear volume (meniscus height and phenol red thread test) and tear stability (noninvasive tear break up time [NI-TBUT]) were significantly reduced in intolerant wearers (P < 0.05). A greater number of symptoms were reported by intolerant than by tolerant wearers (P < 0.05). Tolerance was associated with clinical but not protein characteristics of the tear film. Formulas best able to predict contact lens intolerance included NI-TBUT, number of symptoms experienced, and tear film meniscus height. Formulas had high sensitivity, and specificity which ranged from 29% to 57%.. Contact lens intolerance appears to be best predicted by a combination of clinical variables, including tear film stability, tear volume, and symptom reporting.

Topics: Adult; Conjunctivitis; Contact Lenses; Discriminant Analysis; Dry Eye Syndromes; Eye Proteins; Female; Humans; Immunoglobulin A, Secretory; Keratitis; Lactoferrin; Male; Muramidase; Osmolar Concentration; Patient Satisfaction; Phenolsulfonphthalein; Sensitivity and Specificity; Tears

This study aimed to characterise the composition of the pre-ocular fluid after transplantation of the autologous submandibular gland (SMG) for patients with severe dry eye.. Stimulated and unstimulated pre-ocular fluid from 15 patients (17 eyes) with a viable SMG graft ("SMG-salivary tears"), as well as normal tears and SMG saliva (20 normal subjects/ 20 eyes), was sampled. As global tear parameters, fern pattern analysis and SDS gel electrophoresis were performed. As specific quality parameters, total protein content, secretory immunoglobulin A (SIgA), lysozyme, amylase, sodium, potassium and osmolality were measured using routine laboratory methods. The flow rate of SMG-salivary tears was determined in 5 patients by means of sequential scintillography.. The fern pattern of SMG-salivary tears was coarse and thus more similar to normal SMG saliva than tears. SDS gel electrophoresis of the SMG-salivary tears showed albumin and two unidentified proteins in addition to the normal tear pattern. Osmolality and total protein content of SMG-salivary tears were higher than in normal SMG saliva, but still lower than in normal tears. High activities of normal tear antibacterial proteins (SIgA, lysozyme and amylase) were detected in the salivary tears. Stimulation of the secretion did not alter the composition of SMG-salivary tears. The flow rate of SMG-salivary tears was closer to that of normal tears than normal SMG saliva.. Salivary tears resulting from SMG-transplantation represent condensed SMG saliva. Thus their quality is intermediate between normal tears and normal SMG saliva. High levels of secretory proteins demonstrate that the gland maintains an active function. Surgical denervation and residual tear components from the ocular surface are the most likely factors to cause the complex differences between normal SMG saliva and SMG-salivary tears. The effects of this secretion on the ocular surface are currently being evaluated in a clinical and laboratory study.

Topics: Adolescent; Adult; Aged; Amylases; Dry Eye Syndromes; Female; Humans; Immunoglobulin A, Secretory; Male; Middle Aged; Muramidase; Osmolar Concentration; Potassium; Proteins; Radionuclide Imaging; Sodium; Submandibular Gland; Tears; Transplantation, Autologous; Treatment Outcome

This study aimed to characterise the composition of the pre-ocular fluid after transplantation of the autologous submandibular gland (SMG) for patients with severe dry eye.. Stimulated and unstimulated pre-ocular fluid from 15 patients (17 eyes) with a viable SMG graft ("SMG-salivary tears"), as well as normal tears and SMG saliva (20 normal subjects/20 eyes), was sampled. As global tear parameters, fern pattern analysis and SDS gel electrophoresis were performed. As specific quality parameters, total protein content, secretory immunoglobulin A (SIgA), lysozyme, amylase, sodium, potassium and osmolality were measured using routine laboratory methods. The flow rate of SMG-salivary tears was determined in 5 patients by means of sequential scintillography.. The fern pattern of SMG-salivary tears was coarse and thus more similar to normal SMG saliva than tears. SDS gel electrophoresis of the SMG-salivary tears showed albumin and two unidentified proteins in addition to the normal tear pattern. Osmolality and total protein content of SMG-salivary tears were higher than in normal SMG saliva, but still lower than in normal tears. High activities of normal tear antibacterial proteins (SIgA, lysozyme and amylase) were detected in the salivary tears. Stimulation of the secretion did not alter the composition of SMG-salivary tears. The flow rate of SMG-salivary tears was closer to that of normal tears than normal SMG saliva.. Salivary tears resulting from SMG-transplantation represent condensed SMG saliva. Thus their quality is intermediate between normal tears and normal SMG saliva. High levels of secretory proteins demonstrate that the gland maintains an active function. Surgical denervation and residual tear components from the ocular surface are the most likely factors to cause the complex differences between normal SMG saliva and SMG-salivary tears. The effects of this secretion on the ocular surface are currently being evaluated in a clinical and laboratory study.

Topics: Adolescent; Adult; Aged; Amylases; Dry Eye Syndromes; Electrophoresis, Polyacrylamide Gel; Female; Humans; Immunoglobulin A, Secretory; Male; Middle Aged; Muramidase; Osmolar Concentration; Potassium; Saliva; Sodium; Submandibular Gland; Tears; Transplantation, Autologous

Human tear fluid contains lactoferrin at the highest concentration. In patients with dry eye such as Sjogren's syndrome, the concentration of lactoferrin in the tears is approximately half the normal value. The present study utilizes a short-term rabbit dry eye model to evaluate if lactoferrin containing eye drops can reverse any of the damage produced by blockage of blinking with an ocular speculum. Damage was evaluated based on the extent of methylene blue staining in histological sections. After 3 h of desiccation, the amount of extractable dye recovered following sacrifice increased by more than 4-fold in the vehicle-treated eyes. However, in those rabbits treated with 1% lactoferrin, dye recovery was only 40% of the value in the vehicle-treated eyes. Between 1-3 h and over a concentration range from 0.01 to 1% lactoferrin, the decreases in staining were both time and concentration dependent. Alternatively, if 1% lactoferrin was applied during the desiccation period, there was partial restoration of corneal epithelial integrity. These results suggest that lactoferrin may be of therapeutic value in decreasing the loss of corneal epithelial integrity in dry eye.

Topics: Animals; Coloring Agents; Dry Eye Syndromes; Epithelium, Corneal; Lactoferrin; Male; Methylene Blue; Muramidase; Ophthalmic Solutions; Rabbits; Serum Albumin, Bovine; Staining and Labeling

Despite the high frequency and clinical relevance of blepharitis-associated dry-eye syndrome, no agreement exists about whether diagnostic tests should be performed with or without topical anesthesia. The aim of this study was to compare the influence of topical anesthesia on the mean values of Schirmer's test, tear lysozyme, tear lactoferrin, and tear osmolarity in patients suffering from blepharitis-associated dry eye syndrome.. The authors compared the mean values of Schirmer's test, tear osmolarity, tear lysozyme (turbidimetric assay), and tear lactoferrin (radial immunodiffusion) before and after topical anesthesia in the following groups: 56 normal subjects (group 1), 62 blepharitis patients (group 2), and 15 patients with blepharitis-associated dry eye syndrome (group 3). All clinical and laboratory tests were performed by masked observers.. In group I, mean values of Schirmer's test decreased 24.8% (p < 0.01) when performed after application of topical anesthesia. The other tests were not significantly modified. In groups 2 and 3, significant differences were seen in Schirmer's test (25.33% and 24.19% respectively, p < 0.001) and the lysozyme determination (14.00% and 13.22% respectively, p < 0.01). Differences between the normal subjects (group I) and the patient groups increased when the tests were performed after application of topical anesthesia reaching statistical significance in group 3 for all the tests.. Performing diagnostic tests after topical anesthesia instillation could be useful in detecting dry eye associated with blepharitis.

Topics: Adrenergic alpha-Agonists; Adult; Anesthesia, Local; Anesthetics, Local; Blepharitis; Drug Combinations; Dry Eye Syndromes; Female; Humans; Lactoferrin; Male; Middle Aged; Muramidase; Naphazoline; Ophthalmic Solutions; Osmolar Concentration; Tears; Tetracaine

The lacrimal film contains 6 to 10 g/l of proteins, 99% of which locally synthesized by the lacrimal glands. The study of these proteins allows us to explore the lacrimal function and to reveal an inflammatory process.. The lacrimal proteinic profile included the determination of total proteins and electrophoresis on agarose gel and, if necessary, specific determinations of albumin, lactoferrin, lysozyme and immunoglobulins using a nephelemetric technique. Normal values were established on a hundred of individual tears.. The electrophoretic proteinic profile may present different abnormalities, such as an inflammatory process, a functional alteration of the lacrimal glands or a dysproteinic abnormality of the tears. The specific determination of the principal lacrimal proteins allows us to accurately quantify each of them.. The electrophoresis of the tears on an agarose gel reveals the presence of an inflammatory process or the quantitative or qualitative alteration of the lacrimal function. The immuno-nephelemetric determination of the most important proteins which are involved in theses mechanisms gives an accurate quantitative measurement of proteins and allows biological follow-up of the disease.

Topics: Albumins; Blood-Aqueous Barrier; Conjunctivitis; Dry Eye Syndromes; Electrophoresis, Agar Gel; Humans; Immunoglobulin A; Lacrimal Apparatus; Lactoferrin; Muramidase; Proteins; Tears

Topics: Dry Eye Syndromes; Electrophoresis; Electrophoresis, Paper; Humans; Keratoconjunctivitis; Keratoconjunctivitis Sicca; Lacrimal Apparatus; Muramidase; Proteins; Tears