Emilio C. Campos

Tear proteomics in evaporative dry eye disease

Purpose:To analyze tear protein variations in patients suffering from dry eye symptoms in the presence of tear film instability but without epithelial defects.Methods: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).Results:In EDE patients vscontrols, a significant decrease in levels of lactoferrin (data in %±SD): 20.15±2.64 vs24.56±3.46 (P=0.001), lipocalin-1: 14.98±2.70 vs17.73±2.96 (P=0.0001), and lipophilin A–C: 2.89±1.06 vs3.63±1.37 (P=0.006) was revealed, while a significant increase was observed for serum albumin: 9.45±1.87 vs3.46±1.87 (P=0.0001). No changes for lysozyme and zinc α-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.Conclusions:Data show that tear protein changes anticipate the onset of more extensive clinical signs in early stage dry eye disease.

Subfoveal choroidal blood flow and central retinal function in retinitis pigmentosa

PURPOSE To determine whether subfoveal choroidal blood flow is altered in retinitis pigmentosa (RP) and whether this alteration is associated with central cone-mediated dysfunction. METHODS In 31 RP patients (age range, 15-72 years) with preserved visual acuity (range: 20/30-20/20), subfoveal choroidal blood flow was measured by real-time, confocal laser Doppler flowmetry, and focal macular (18°) electroretinograms (FERGs) were elicited by 41 Hz flickering stimuli. Twenty normal subjects served as controls. The following average blood flow parameters were determined based on three 60-second recordings: volume (ChBVol), velocity (ChBVel), and flow (ChBF), the last being proportional to blood flow if the hematocrit remains constant. The amplitude and phase of the FERG first harmonic component were measured. RESULTS On average, ChBF and ChBVel were reduced by 26% (P ≤ 0.02) in RP patients compared to controls, whereas ChBVol was similar in the two groups. FERG amplitudes were reduced by 60% (P < 0.01) in patients compared with controls. FERG phases of patients tended to be delayed (P < 0.08) compared with their values in the controls. In patients, FERG phase delays were correlated (r = 0.50, P < 0.01) with ChBF and ChBVel values. FERG amplitudes were correlated (r = 0.49, P < 0.01) with ChBVol values. CONCLUSIONS These data indicate significant alterations of subfoveal choroidal hemodynamic in RP and suggest a link between the alteration of ChBF and the RP-associated central cone-mediated dysfunction as assessed by the FERG.

Pseudoexfoliation Syndrome: A Cohort Study on Intraoperative Complications in Cataract Surgery

Our aim was to determine the rate of intraoperative complications induced by pseudoexfoliation syndrome (PSX) in cataract surgery. We report our experience regarding 1,052 consecutive patients who underwent phacoemulsification. Exact logistic regression was used to examine the role of PSX in intraoperative complications. Odds ratio for intraoperative complications (vitreous loss, capsular break, zonular break) was estimated to be 5.1 for PSX present as compared to when it is absent. PSX was associated with a statistically significant increase in intraoperative complications during cataract surgery (p < 0.0001).

Effect of citicoline on visual acuity in amblyopia: preliminary results

Abstract•Background: Citicoline has been used to improve consciousness levels in cerebral trauma and as a complement for levodopa in Parkinsons disease. It has also been shown that severe glaucomatous visual field deficits improve for at least 4 months with the use of citicoline. In this paper, preliminary results are presented of an open and a double-blind study on the effect of citicoline in amblyopia.•Methods: The open study was started in 1991. Fifty patients with amblyopia were treated with citicoline (1000 mg i.m. daily for 15 days). They were selected from an age group beyond the plastic period of the visual system. Occlusion or other types of anti-amblyopic treatment were never used at the same time as citicoline. A double-blind study was conducted on 10 more patients, randomly assigned to a treatment or placebo group and followed for 6 months. A statistical test for repeated measures was performed on all the results.•Results: A statistically significant improvement in visual acuity was found both for the amblyopic and the sound eye in 46 of the 50 patients (92%). The behaviour was different for normal and amblyopic eyes. The improvement remained stable for at least 4 months. These results have been substantiated by the double-blind study. No side effects were observed.•Conclusion: Citicoline improves visual acuity, at least temporarily, in amblyopic patients beyond the plastic period of the visual system. Our results suggest that trials of citicoline as a medical treatment of amblyopia are warranted.

Diagnostic performance of tear function tests in Sjogren's syndrome patients

ObjectiveTo evaluate the diagnostic performance of the tests included in primary Sjogrens syndrome (SS-I) diagnostic criteria (Schirmer I, break-up time, vital dye staining) and to compare them with other examinations related to the ocular surface status.MethodsClinical and cytological data were collected from 177 patients (62 SS-1, 56 non-SS autoimmune diseases, 59 Sicca syndrome). Tear tests included: a validated questionnaire on symptoms, Schirmer I, Jones test, Ferning test, BUT, corneal aesthesiometry, tear clearance test, lissamine green staining, impression conjunctival cytology. Data were statistically evaluated and sensitivity, specificity, likelihood ratio (LR+), receiver-operating characteristics (ROC) curves were calculated for each test.ResultsData showed a poor diagnostic performance of Schirmer test I (sensitivity 0.42; specificity 0.76; LR+1.75) and BUT (sensitivity 0.92; specificity 0.17; LR+1.11) (area under the curve in ROC analysis <0.58). Validated subjective symptoms questionnaire (sensitivity 0.89; specificity 0.72; LR+3.18), Jones test (sensitivity 0.60; specificity 0.88; LR+5), corneal aesthesiometry (sensitivity 0.80; specificity 0.67; LR+2.42), and tear clearance test (sensitivity 0.63; specificity 0.84; LR+3.93), all exhibited a high diagnostic performance (area under the curve in the ROC analysis always >0.70). Lissamine green staining exhibited the best performance (sensitivity 0.63; specificity 0.89; LR+5.72) but the result could be distorted by an incorporation bias.ConclusionsOur data suggest to implement the items for ocular signs and symptoms contained in many SS-I diagnostic criteria with the use of a validated questionnaire, performance of Jones test, corneal aesthesiometry measurement, and tear clearance rate evaluation.

Menopause and dry eye. A possible relationship

The hormonal status and in particular sex steroids play a role in the ocular surface homeostasis and function, during the whole life and in both sexes, accomplished by estrogenic and androgenic receptors located on corneal and conjunctival epithelia and Meibomian gland. The ocular surface is an integrated unit and any dysfunction results in a scarce or unstable preocular tear film which produces dry eye, a chronic inflammatory condition with increasing incidence as people get older but always more frequent among females. In post-menopausal women endocrine changes join the aging effects in the pathogensis of dry eye, but still it remains controversial whether estrogen or androgen deficiency or their imbalance impair ocular surface function. Another questionable issue concerns the efficacy of hormonal replacement therapy in the amelioration of dry eye symptoms and recovery of tear function, since the scientific literature stands in between a therapeutic or a promoting effect of eye dryness. Therapy of dry eye is usually based upon topical administration of tear substitutes; estrogen or androgen-based eye drops represents a promising innovative treatment based upon important scientific rationale.

Hyperosmolar Stress Upregulates HLA-DR Expression in Human Conjunctival Epithelium in Dry Eye Patients and In Vitro Models

PURPOSE To investigate the immune response of human conjunctival epithelium to hyperosmolar stress. METHODS Tear osmolarity was measured in 15 normal subjects and 25 dry eye (DE) patients; conjunctival imprint cytology samples were obtained at the nasal bulbar area. Subconfluent primary human conjunctival epithelial cells (pHCECs) and human conjunctival organ cultures (hCOCs), both cultured in iso-osmolar medium (305 mOsm/L), were exposed for 24 hours to media with progressively higher osmolarity, with or without the ion channel inhibitor ruthenium red (RuR). Human leukocyte antigen (HLA)-DR expression was evaluated by immunocytochemistry, on imprints from subjects, on primary human conjunctival epithelial cells, on formalin fixed-paraffin embedded hCOCs, and by RT-PCR. Statistical evaluation was performed by applying the unpaired Students t test, as well as Spearmans rho and Pearsons r correlation coefficients (significance P < 0.05). RESULTS HLA-DR expression increased in DE subjects with respect to control (% mean ± SD, respectively, 46.16 ± 7.2 vs. 7.48 ± 1.14, P < 0.0001) and exhibited significantly high correlations with tear osmolarity values (r = 0.614; P < 0.0001). In vitro experiments showed a progressive increase in HLA-DR expression as the osmolarity of the medium was increased from 6.75 ± 1.16 (% mean ± SD) in iso-osmolar-cultured cells to 9.96 ± 1.37 and 12.94 ± 4.04 in cells cultured in, respectively, 350 and 400 mOsm/L (P < 0.05). A stepwise progressive increase was also found in hCOCs. Results were confirmed by RT-PCR. Ruthenium red significantly reduced HLA-DR expression in hyperosmolar-cultured cells. CONCLUSIONS Data from complementary techniques demonstrate that extracellular hyperosmolarity induces HLA-DR overexpression in human conjunctival epithelial cells in both DE patients and in vitro cell culture models.

Amount of Aniseikonia Compatible with Fine Binocular Vision: Some Old and New Concepts

Random Dot Stereograms (RDS) are useful, but are sometimes misleading in assessing the degree of binocular cooperation in the presence of aniseikonia. RDS may give positive results up to 15% aniseikonia, whereas a fine central binocularity is not possible with more than 5% image-size difference. It is suggested that stereopsis may not be the only parameter to test; stress is given to the importance of peripheral motor fusion in maintaining the eye alignment.

Prostaglandin analogs and blood-aqueous barrier integrity: a flare cell meter study.

Purpose: To study, with an objective method, inflammation of the anterior segment of the glaucomatous eye after treatment with latanoprost, travoprost and bimatoprost. Materials and Methods: Sixty patients with chronic open-angle glaucoma aged between 38 and 76 years (mean 64.0 ± 12.2) were randomly assigned to latanoprost 0.005, travoprost 0.004 and bimatoprost 0.03%. The study period lasted 6 months. Intraocular pressure (IOP) was measured every 2 weeks. We studied the intraocular inflammation before and after 3 and 6 months of therapy with an instrument composed of a He-Ne laser beam system, a photomultiplier mounted on a slitlamp microscope and a computer. This flare meter allows objective determination of the flare and the number of cells in the aqueous of the anterior chamber. Results: At the baseline, IOP was 26.4 ± 3.6 mm Hg. After 3 months of treatment, mean IOP in the latanoprost group was 17.9 ± 0.3 mm Hg (p < 0.001) with a mean cellularity of 12.638 ± 3.284 photons/ms (p < 0.001). The travoprost group had an IOP of 17.2 ± 0.3 mm Hg (p < 0.001) with a cellularity of 9.719 ± 1.927 photons/ms (0.001). Finally, IOP in the bimatoprost group was 17.6 ± 0.5 mm Hg (p < 0.001) with a cellularity of 6.138 ± 1.475 photons/ms (p < 0.032). After 6 months of treatment, IOP in the latanoprost group was 18.1 ± 0.3 (p < 0.001), in the travoprost group 17.3 ± 0.3 (p < 0.001) and in the bimatoprost group 17.7 ± 0.5 mm Hg (p < 0.001), whereas cellularity was 11.838 ± 3.218 (p < 0.001), 8.950 ± 3.692 (p < 0.001) and 7.617 ± 2.603 photons/ms (p < 0.001), respectively. After 3 months, the travoprost (p < 0.013) and the bimatoprost groups (p < 0.001) had less flare compared with the latanoprost group and this remained so even at 6 months. When we compared the travoprost group with the bimatoprost group, we found significantly less flare at 3 months in the bimatoprost group (p < 0.001) but not at 6 months (p < 0.246). Conclusions: The flare meter analysis shows that the eyes treated with bimatoprost and travoprost have a less significantly broken blood-aqueous barrier and their anterior chamber is also significantly less inflamed.

Efficacy of standardized and quality-controlled cord blood serum eye drop therapy in the healing of severe corneal epithelial damage in dry eye.

Purpose: We standardized quality-controlled cord blood serum (CBS)–based eye drops and evaluated the efficacy of 1-month CBS treatment in the healing of diseased corneal epithelium in severe dry eye (DE) patients. Methods: Seventeen graft-versus-host disease (GVHD) and 13 Sjogren syndrome patients with severe persistent corneal defects were enrolled in the framework of a registered clinical trial (ClinicalTrials.gov NCT01234623). Sterile CBS eye drops were prepared to supply 0.15 ng per eye per day epithelial growth factor and administered for 1 month in a 1-day dose dispensing. The extent of epithelial defect was evaluated in square millimeters area, and subjective symptom score (Ocular Surface Disease Index score), Schirmer test I, break-up time, tear osmolarity, corneal esthesiometry (Cochet–Bonnet esthesiometer), conjunctival scraping, and imprint cytology with goblet cell count were performed at baseline (V0) and after 15 (V1) and 30 (V2, endpoint) days of treatment. Satisfaction and tolerability questionnaires were evaluated at V1 and V2. Results: A significant reduction was shown at the endpoint versus baseline in corneal epithelial damage (mean ± SD, 16.1 ± 13.7 vs. 40.9 ± 30 mm2/area, respectively), discomfort symptoms (Ocular Surface Disease Index score, 22.3 ± 10.3 vs. 39.3 ± 16.9), scraping cytology score (3.8 ± 1.2 vs. 6.6 ± 2.1), and tear osmolarity (312.5 ± 7 vs. 322 ± 9.1 mOsm/L), whereas a significant improvement was shown in corneal esthesiometry (48.2 ± 2.1 vs. 49.7 ± 2.1 nylon/mm/length, P < 0.05). All patients reported a high degree of satisfaction upon drop instillation. Conclusions: Heterologous CBS-based eye drops represent a promising therapeutic approach in the healing of severely injured corneal epithelium and in subjective symptom relief. These drops can be obtained as readily available and quality-controlled blood derivative from cord blood banks on a routine basis.