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PubMed2000

Tear levels of interferon-gamma, interleukin (IL) -2, IL-4 and IL-5 in patients with vernal keratoconjunctivitis, atopic keratoconjunctivitis and allergic conjunctivitis.

BACKGROUND: A predominance of TH2 activity in chronic allergic diseases, such as vernal keratoconjunctivitis (VKC) and atopic keratoconjunctivitis (AKC), has been suggested recently. However, there is no published study on tear levels of cytokines of the two different subgroups, TH1 and TH2, in patients with ocular allergy. OBJECTIVES: We measured interferon (IFN)-gamma, interleukin (IL)-2, IL-4 and IL-5 levels in tears by ELISA, to determine whether the levels of these cytokines are elevated in allergic ocular diseases when compared among patient groups and normal controls. METHODS: Tear levels of IL-2, IFNgamma, IL-4 and IL-5 were measured by ELISA using samples from patients with VKC, AKC (AKC-NP, without proliferative lesions; and AKC-P, with proliferative lesions), allergic conjunctivitis (AC) and normal subjects. The levels of these cytokines in tears and the clinical severity of AD were also compared. RESULTS: Tear IL-4 level in patients with AKC was significantly higher than those in VKC, AC and controls, and tear IL-4 levels in patients with AKC-P vs VKC differed significantly. Tear IL-5 levels in patients with diseases associated with proliferative lesions, VKC and AKC-P, were higher than those in AC and normal controls. However, tear level of IL-5 in patients with AKC-P was significantly higher than that in AKC-NP. Although the dermatological severity of AD correlated significantly with tear IL-4 level, IFNgamma, IL-2 and IL-5 levels did not correlate with dermatological severity of AD. CONCLUSION: These results indicate that the TH2-like cytokines play an important pathophysiological role in severe ocular allergic conditions such as AKC and VKC and that tear level of IL-5 may be a candidate marker to evaluate the clinical status of ocular allergy. The different patterns of tear levels of IL-4 and IL-5 among ocular allergic diseases may reflect the origin and immunological basis of these cytokines.

Adolescent↗
PubMed1998

Fluorophotometric measurement of the buffering action of human tears in vivo.

PURPOSE: The buffering action of human tears is thought to be important to keep its pH constant. We measured the change in pH in the precorneal tear film in vivo when the acidic solution is challenged, using a fluorophotometric technique. METHODS: Twelve eyes from 6 healthy subjects were entered in this study. Each subject was pretreated with either one drop of 0.4% oxybuprocaine for once (light anesthesia), three times (deep anesthesia), or none (controls). The measurement was initiated by instilling 20 microl of 0.067 M phosphate buffer at pH 5.5 containing 2 mM bis-carboxyethyl-carboxyfluorescein free acid, a pH sensitive dye, into the subject's eye. The pH was determined by the ratio of fluorescent intensities at two excitation wavelengths (490 and 430 nm). pH recovery time (PHRT) as defined by the time required for pH to reach 95% of pH at equilibrium was used for the marker of tear buffering action. Tear turnover rate was also determined using the fluorescent decay curve at 430 nm, which was independent of pH, but dependent on dye concentration. RESULTS: Immediately after the instillation, the pH value in the tear film was around 6.0-6.5 in all cases. The tear film rapidly became more alkaline, reaching its normal value in 2.3 +/- 0.5 min in untreated eyes. The pretreatment with 0.4% oxybuprocaine retarded the neutralization process. A single regression analysis revealed that the PHRT had a significant negative correlation with the tear turnover rate (r = -0.78). CONCLUSIONS: Our results suggest that the neutralization process of tears largely depends on the tear turnover rate. The buffering action of tears in vivo consists of the tear turnover as well as its chemical buffering capacity.

Acid-Base Equilibrium↗
PubMed1999

Components responsible for the surface tension of human tears.

PURPOSE: It was previously thought that the surface tension of tears was due to dissolved mucin, but it has recently been shown that very little mucin is present. The surface tensions of solutions of commercial mucin, lysozyme, lactoferrin or secretory IgA are all higher than that of tears. The influence of tear lipocalin and lipids remained to be tested. METHODS: Surface tension was determined by a micro-method on pooled intact stimulated human tears, and following extraction with lipid solvents. The extracted material was also added back, as was a variety of lipid standards (phospholipids, glycolipids, sterols, etc.). TLC and GLC were used in partial identification of the extract. Another lipocalin, bovine beta-lactoglobulin, was also tested alone and mixed with tear lipids, model lipids, or model tear proteins. RESULTS: Intact tears had a surface tension of 42-46 mN/m, but after lipid extraction this rose to 53-55.5 mN/m. Addition of lipids to the delipidised tear fluid gave a range of tensions from 42 to 49 mN/m, with the greatest effects shown by phospholipids (phosphatidylcholine, sphingomyelin), but full recovery was only achieved by using the extracted lipid material. Human meibomian oil was less effective. The GLC peak profile of the extract was markedly different from meibomian oil, and the TLC pattern was consistent with the presence of glycolipids. CONCLUSIONS: The surface tension of tears is due to a complex of tear lipocalin with a polar lipid fraction extractable from tears by lipid solvents and different from meibomian lipid. Lipocalin and this lipid fraction may be secreted together by the lacrimal gland.

Centrifugation↗
PubMed1999

Tear lysozyme activity in frozen Schirmer strips and salivary gland biopsy as parameters of lacrimal gland function.

PURPOSE: The lysozyme concentration in human tears is an important parameter for tear gland function. The decline of lysozyme in tears reflects lacrimal gland destruction. In Sjögren's patients, lacrimal gland destruction parallels labial salivary gland destruction. The objective of this study was to determine whether human tear lysozyme that was frozen on Schirmer strips at -20 degrees C for several years maintained activity and whether there was a linear relation with inflammatory changes in labial salivary glands. METHODS: A total of 200 frozen Schirmer strips were processed. They were collected from 20 randomly selected patients each year of five consecutive years, all attending the UCSF Sjögren's Clinic. The tear lysozyme in the Schirmer strips was measured by a colorimetric assay. The average lysozyme concentration each year was calculated and compared. One third of the patients underwent labial salivary gland biopsy. The correlation was calculated between the tear lysozyme concentration and the lymphocytic focus scores in biopsy specimens. RESULTS: No significant difference of average lysozyme concentration in the Schirmer strips was found when the five different years of collection were compared. The linear relation between the tear lysozyme concentrations and the focus score in labial salivary gland biopsies showed a coefficient of r = -0.41. The linear relation between other diagnostic measurements, like Schirmer test, tear breakup time, or rose bengal staining pattern, and the focus score was lower. CONCLUSIONS: Human tear lysozyme in Schirmer strips can be stored at -20 degrees C for at least five years. There is little difference in lysozyme activity of frozen compared to unfrozen specimens. The lysozyme concentration in tears correlates better with the lymphocytic focus score in labial salivary gland biopsy than does clinical assessment and is therefore a parameter for the actual degree of tear gland destruction.

Biopsy↗
PubMed1996

Tear film changes associated with normal aging.

We examined a group of normal subjects to evaluate the tear film function by measuring reflex tear capacity using the Schirmer test without anesthetic, steady state tear flow using fluorophotometry, meibomian gland function based on gland drop-out, expressible lipid volume and viscosity, tear loss from evaporation, and tear osmolarity. We found the average tear flow to be 0.19 +/- 0.19 microliters/min, lower than previously measured. The average tear volume was also correspondingly less, 2.74 +/- 2.0 microliters. We found there was a decline associated with advancing age for all parameters except the decay constant and lipid volume. The highest correlations with aging were tear volume (-0.64), Schirmer test (-0.63), and tear osmolarity (0.59). Our results indicate that there is a decline in tear film function throughout life and that tear flow is considerably less than previously estimated.

Adolescent↗
PubMed2003

Use of retroillumination to visualize optical aberrations caused by tear film break-up.

PURPOSE: The aim of the current study was to develop quantitative methods to assess optical aberrations caused by tear film disruption. METHODS: We used standard fluorescein imaging (FL) and a novel retroillumination (RI) method to image tear film disruption in 12 eyes. Using a clinical slit lamp biomicroscope, we alternated between widefield blue and narrow-beam white light to obtain an interleaved series of FL and RI images of the time course and pattern of tear film break-up. We developed an optical analysis that indicates that the RI image should be proportional to the spatial derivative of the FL image. Intensity fluctuations in the RI images are due to thickness changes in the tear film, whereas intensity fluctuations in FL images are directly determined by tear film thickness. RESULTS: As predicted by optical analysis of RI, the spatial distribution of gaps in the tear film seen with fluorescein appeared as pairs of light and dark contours in the RI images, and a precise correspondence between the spatial derivative of the FL image (slope) and the RI image was found. Both methods showed a gradual spreading of the tear disruption during blink suppression that varied tremendously among eyes in both time and spatial pattern. Resumption of normal blinking did not produce an immediate reconstitution of the normal tear film, and areas of tear break-up created during blink suppression remained abnormal for up to several minutes of normal blinking. CONCLUSIONS: Our analysis indicates that both FL and RI have the potential to quantify optical changes occurring during tear break-up. These results support an interpretation of RI as an intensity-based method for mapping the highly irregular optical aberrations of the eye produced by tear film disruption.

Adult↗
PubMed2003

The effect of artificial tears on visual performance in normal subjects wearing contact lenses.

PURPOSE: Disruption of the anterior refracting surface of the eye reduces visual performance. In the case of a contact lens wearer, this surface is the prelens tear film. This study determined whether artificial tears stabilize the tear film in contact lens wearers and maintained optimal visual performance for a prolonged period. METHODS: Five normal subjects all in good general and ocular health and adapted to contact lens wear were fitted with Acuvue lenses. A temporal, two-alternative, forced-choice paradigm was used to measure contrast sensitivity. The stimuli were vertically oriented sine-wave gratings (between 0.5 and 14 cpd) presented for 16.67 ms. The stimuli were presented at two different times after blink detection: 2 s after blink detection (i.e., before tear layer break-up) or 4 s after tear film break-up. Three conditions were investigated at 4 s after tear layer break-up: (1) without artificial tears added, (2) with Clerz2 (Ciba Vision) instilled, and (3) with Sensitive Eyes (Bausch & Lomb) applied. The artificial tears were instilled at 10-min intervals during the experimental run. RESULTS: High spatial frequency contrast sensitivity was found to be reduced after tear film break-up and was not enhanced by either tear solution. CONCLUSIONS: This study indicates that there is no effect of Clerz2 or Sensitive Eyes in maintaining or improving visual performance beyond the normal prelens tear film break-up time in normal subjects wearing Acuvue lenses.

Adult↗
PubMed2004

Effect of contact lens materials on tear physiology.

PURPOSE: This study measured evaporation rate, thinning characteristics, and lipid layer changes in the prelens tear film (PLTF) associated with wearing of different soft contact lens materials, in an attempt to determine the biocompatibility of the material with the PLTF. METHODS: Twenty habituated contact lens wearers wore five different soft materials in a random order on the left eye at visits separated by at least 24 h. The soft contact lens materials were polymacon (Optima 38), omafilcon A (Proclear Compatibles), phemfilcon A (DuraSoft 2), balafilcon A (PureVision), and etafilcon A (Acuvue). Tear film evaporation rate was measured by a modified Servo Med Evaporimeter and tear thinning time by HirCal grid. Tear film structure, elimination rate, and lens wetting ability were recorded dynamically with a Doane tear film video interferometer and graded according to a new system developed for the study. Baseline measurements were taken of the precorneal tear film before lens insertion, and PLTF was determined 30 min after commencing lens wear. RESULTS: No statistically significant differences were found for any of the baseline (precorneal tear film) data. There was also no significant difference in evaporation rate change (analysis of variance) and in tear thinning time (Friedmann) between the five contact lenses. In the PLTF structure grading, omafilcon A had significantly more stable grades than phemfilcon A (Friedmann, p = 0.0033) and polymacon (p = 0.004). In PLTF observation of tear thinning and elimination rate, there was a significantly slower rate of elimination observed for omafilcon A than phemfilcon A (Friedmann, p = 0.0023) and polymacon (p = 0.0023). There was no significant difference in the overall PLTF wetting ability grading between any of the lenses worn. CONCLUSION: Generally, all soft contact lens materials significantly and adversely affected tear physiology by increasing the evaporation rate and decreasing tear thinning time. The surface wetting ability of all contact lens materials exhibited no significantly difference irrespective of the special surface treatments. Only in PLTF structure and in PLTF elimination rate were differences found from the conventional low water content materials; omafilcon A was better in PLTF structure and in PLTF elimination.

Adolescent↗
PubMed1999

Acetaminophen (paracetamol) levels in human tears.

This study was designed to measure acetaminophen (paracetamol) levels in tears, and to compare it to serum levels. Paracetamol levels were measured in 20 paired tears and serum samples from 10 healthy volunteers, 1 and 2 hours after ingesting 1.5 g paracetamol. Tears were collected using glass microcapillary tubes while stimulating the conjunctiva with a small sponge placed in the lower fornix. Blood samples were taken simultaneously. The samples were analyzed for paracetamol levels using homogeneous enzyme immunoassay. Tears and serum paracetamol levels 1 hour after ingestion were 16.3 microg/mL +/- 7.2 (mean +/- SD), and 21.4 microg/mL +/- 7.7 (mean +/- SD) respectively. Tears and serum levels 2 hours after ingestion were 14.4 microg/mL +/- 7.8 (mean +/- SD), and 17 microg/mL +/- 7.6 (mean +/- SD) respectively. Tears and serum paracetamol levels of all the 20 paired samples (1 h and 2 h after ingestion) were 15.35 microg/mL +/- 7.4, and 19.25 microg/mL +/- 7.8, respectively (mean +/- SD). There was a strong and highly significant correlation between paracetamol levels in serum and in tears 1 and 2 hours after ingestion (r = 0.8, p = 0.005, r = 0.85, p = 0.002 respectively). Mean +/- SD ratio of tears/serum paracetamol levels 1 hour and 2 hours after ingestion were 0.77 +/- 0.21 and 0.81 +/- 0.25 respectively. Delta tears (difference in mean levels at 1 and 2 hours) paracetamol levels is significantly correlated with delta serum levels (r = 0.7, p = 0.025). A reliable, convenient, and feasible noninvasive method is described for measuring paracetamol in tears. There is no information in the literature about tears paracetamol secretion, and little information of tears drugs concentration.

Acetaminophen↗
PubMed2005

Effect of experimental dry eye on tear sodium concentration in the mouse.

PURPOSE: To examine the changes in tear volume, sodium concentration, and osmolarity in BALB/c and C57BL/6J mice in response to experimental dry eye (EDE). METHODS: Tear volumes were determined at baseline and after 2 days of EDE using phenol red-impregnated cotton thread. Tear fluid was collected by instilling 1 microl of distilled water into the conjunctival sac and collecting the diluted tear fluid in a 0.5-microL glass capillary tube. CoroNa Red in dimethyl sulfoxide DMSO was added, and the fluorescent intensity was measured, which was compared to a standard curve of sodium concentrations. The resulting concentration was then corrected by the proper dilution factor. The tear osmolarity owing to sodium was calculated by comparing the calculated tear sodium concentration to a standard curve established by a vapor pressure osmometer. Serum osmolarity was also determined using a vapor pressure osmometer. RESULTS: After 2 days of EDE, tear volume significantly decreased from 0.093 microL to 0.028 microL in BALB/c mice and from 0.066 microL to 0.026 microL in C57BL/6J mice. There was a concomitant significant increase in tear osmolarity from 177 mOsm/L to 300 mOsm/L in C57BL/6J mice. Tear osmolarity nearly doubled from 285 mOsm/L to 559 mOsm/L in BALB/C mice with EDE, approaching statistical significance (P=0.12). No change in the serum osmolarity was observed in either mouse strain. CONCLUSIONS: These experiments show the ability of these new techniques for determining tear volume and estimating tear osmolarity in mice. By mimicking the findings of human dry eye disease, these findings validate the relevance of the mouse model for studying the pathogenesis of human keratoconjunctivitis sicca.

Animals↗
PubMed1996

Detection of allergen-specific IgE in tears of grass pollen-allergic patients with allergic rhinoconjunctivitis.

BACKGROUND: Allergic conjunctivitis is a common symptom amongst Type I (IgE-mediated) allergic diseases; and most frequently seen as rhinoconjunctivitis. However, the site of production and the significance of allergen specific IgE needs further elucidation. OBJECTIVE: We investigated whether the presence of IgE in tears of grass pollen allergic patients correlated with disease and clinical symptoms, whether the IgE binding pattern to the different grass pollen antigens was different in sera and tears, and whether IgA antibodies to grass pollen allergens were present in tears. Finally, we looked whether specific IgE was produced locally or was exudated from serum. METHODS: Sera from 44 grass pollen allergic patients suffering from either allergic rhinitis (n = 11) or allergic rhinoconjunctivitis (n = 33) and from healthy controls (n = 7) were used for the experiments. Binding of specific IgE and IgA antibodies to the different groups of grass pollen allergens (Phleum pratense) was evaluated by means of immunoblotting. RESULTS: Specific IgE was detected in sera as well as in tears of allergic patients, whereby tear-derived allergen-specific IgE exerted similar specificities to the corresponding IgE from serum. The correlation between symptoms of ocular allergy and the presence of allergen-specific IgE in tears was highly significant (P < 0.0001). In contrast, only a poor correlation was found between specific and/or total IgE in sera and the manifestation of ocular allergy (P = 0.73). CONCLUSION: Allergen-specific IgE antibodies in tears seem to be produced locally rather than exudated from serum. IgE in tears seems to be responsible for allergic conjunctivitis. IgA in tears cannot exert a protective function since the IgA antibodies recognize different antigens in a grass pollen (Phleum pratense) extract than IgE antibodies. The highly significant correlation between allergic conjunctivitis and the presence of specific tear IgE emphasizes the diagnostic value of immunoblots with tear IgE, especially in cases in which serum provides inconclusive results.

Allergens↗
PubMedDate not supplied

Tear urea nitrogen and creatinine levels in horse and their correlation with serum values.

The objective of this paper was to determine the physiological values of urea nitrogen and creatinine in tears, and to compare the results with those obtained from serum. Thirty healthy thoroughbred horses were included in the study. Tear fluid samples were obtained using a glass capillary tube placed in lower conjunctival cul-de-sac. Blood samples were taken from the jugular vein. Tear and serum urea nitrogen and creatinine levels were quantitatively analyzed by an enzymatic colorimetric method. Urea nitrogen values were 4.22+/-1.84 mmol/l in tears and 4.44+/-1.78 mmol/l in serum, whereas creatinine values in tears were 14.14+/-7.74 micromol/l and in serum 147.63+/-12.17 micromol/l. Statistical analysis confirmed a significant correlation between serum and tear urea levels (P<0001). However, there was no significant correlation between blood and tear creatinine values. Mean value of creatinine obtained from tears was 9.6% of the mean value from serum. Urea nitrogen and creatinine levels can be measured in tears. A significant correlation was found between serum and tears urea levels. This finding may permit development of a new alternative laboratory diagnosis of uremia based on the content of urea in tears.

Animals↗
PubMed2005

Surfactant protein D is present in human tear fluid and the cornea and inhibits epithelial cell invasion by Pseudomonas aeruginosa.

We have previously shown that human tear fluid protects corneal epithelial cells against Pseudomonas aeruginosa in vitro and in vivo and that protection does not depend upon tear bacteriostatic activity. We sought to identify the responsible tear component(s). The hypothesis tested was that collectins (collagenous calcium-dependent lectins) were involved. Reflex tear fluid was collected from healthy human subjects and examined for collectin content by enzyme-linked immunosorbent assay (ELISA) and Western blot with antibody against surfactant protein D (SP-D), SP-A, or mannose-binding lectin (MBL). SP-D, but not SP-A or MBL, was detected by ELISA of human reflex tear fluid. Western blot analysis of whole tears and of high-performance liquid chromatography tear fractions confirmed the presence of SP-D, most of which eluted in the same fraction as immunoglobulin A. SP-D tear concentrations were calculated at approximately 2 to 5 microg/ml. Depletion of SP-D with mannan-conjugated Sepharose or anti-SP-D antibody reduced the protective effect of tears against P. aeruginosa invasion. Recombinant human or mouse SP-D used alone reduced P. aeruginosa invasion of epithelial cells without detectable bacteriostatic activity or bacterial aggregation. Immunofluorescence microscopy revealed SP-D antibody labeling throughout the corneal epithelium of normal, but not gene-targeted SP-D knockout mice. SP-D was also detected in vitro in cultured human and mouse corneal epithelial cells. In conclusion, SP-D is present in human tear fluid and in human and mouse corneal epithelia. SP-D is involved in human tear fluid protection against P. aeruginosa invasion. Whether SP-D plays other roles in the regulation of other innate or adaptive immune responses at the ocular surface, as it does in the airways, remains to be explored.

Animals↗
PubMed2005

Tear film dynamics in floppy eyelid syndrome.

PURPOSE: Floppy eyelid syndrome (FES) presents nonspecific ocular surface irritation. The hypothesis for the current study was that one contributing factor is the abnormality in tear film dynamics. METHODS: Sixteen patients with FES were consecutively examined. Tear film dynamics were evaluated by kinetic tear interference images, infrared thermometry, water evaporation rate, tear break-up time, and fluorescein clearance test. Data showing evaporation rate and thermometry were compared with those of 10 normal subjects. RESULTS: There was a high correlation between the eye with the worse symptoms and the eyes with the more severe floppy lids (P < 0.01) and with ocular surface evaporation rate (P = 0.02). Except for one patient, all others showed abnormal tear film, with an average tear break-up time of 2.9 +/- 3.7 seconds. Kinetic analysis of tear interference images revealed that lipid spread in a vertical or mixed pattern in 18 eyes (75%) with a delayed spread time (P = 0.0007), indicating that most of the patients had lipid tear deficiency. The ocular skin temperature and water evaporation rate were higher in the FES group (P = 0.0003 and 0.026, respectively). Nearly all patients with FES showed eyelid hyperpigmentation. The ocular surface evaporation rate in the FES group was also higher than that of the normal subjects (P < 0.0001). Multiple regression analysis showed that a vertical pattern of lipid spread had a significant influence on ocular surface evaporation rate (P = 0.003). CONCLUSIONS: Tear film abnormality is prevalent in patients with FES and is characterized by lipid tear deficiency, leading to rapid tear evaporation. The FES lid skin is also characterized by high temperature, high water evaporation rate, and hyperpigmentation. Studies directed to investigating the linkage of lid changes and meibomian gland dysfunction may shed new lights on the pathogenesis of FES.

Adult↗
PubMed2005

Thinning rate of the precorneal and prelens tear films.

PURPOSE: To apply interferometry as an in vivo measure of tear film thinning between blinks. METHODS: Wavelength-dependent interference was used to measure the tear film thinning rates in 20 normal contact lens wearers, and spectra were captured at a rate of 4.5 per second for 20 seconds. Four recordings of precorneal tear film (PCTF) thinning were made, followed by 1 hour of hydrogel lens wear and then four recordings of prelens tear film (PLTF) thinning. Subjects were asked to blink 1 second after the beginning of the recording and then hold their eyes open for an additional 19 seconds, followed by 2 minutes of rest between recordings. RESULTS: The average thinning rate of the PLTF was greater than that of the PCTF and average initial tear film thickness of the PLTF was less than that of the PCTF. For both these reasons, the "tear thinning time" (time to reach 0 thickness) was typically shorter for the PLTF than for the PCTF. Histograms of PCTF and PLTF thinning rates showed a narrow peak corresponding to slow thinning of approximately 1 microm/min, but also many examples of rapid thinning of approximately 10 microm/min, with greater variability. Both the initial thickness and thinning rate of the PLTF correlated with corresponding values for the PCTF, although many more rapidly changing values were associated with the PLTF. Plots of rapid thinning of PCTF and PLTF were both linear and were not accompanied by any significant increase in thickness of corneal epithelium or contact lens, respectively. CONCLUSIONS: Tear film thinning can be analyzed in terms of flow in three spatial directions: (1) outward (evaporation), (2) inward (into the epithelium or contact lens), and (3) parallel to the tear film surface. The results indicate that the second mechanism may be unimportant. Studies have shown a range of tear film evaporation rates from 0.24 to 1.45 microm/min, whereas, when the lipid layer is washed away from the tear film, the thinning rate, due to evaporation, would be approximately 7 microm/min. Thus, slow thinning rates may be due to tear film evaporation, whereas rapid rates (which are often greater than 7 microm/min) presumably include other mechanisms such as dewetting, Marangoni flow (i.e., surface tension gradients), and pressure-gradient flow.

Adult↗
PubMed2006

Identification and comparison of the polar phospholipids in normal and dry eye rabbit tears by MALDI-TOF mass spectrometry.

PURPOSE: To identify and compare the phosphorylated lipids in normal and dry eye rabbit tears using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). METHODS: MALDI-TOF MS studies were performed on tear samples from normal and dry eyes of female New Zealand White rabbits. Experimental dry eye was induced by complete removal of the main and accessory lacrimal glands and nictitating membranes. A solid ionic crystal MALDI matrix of paranitroaniline and butyric acid was used to enhance the mass spectral responses of the phospholipids. In addition, a novel lipid isolation, preconcentration, and clean-up method using pipettes containing immobilized metal ion affinity chromatography (IMAC) medium was used. RESULTS: The polar phospholipids present in the normal and dry eye rabbit tears showed both similarities and differences. Species related to platelet-activating factor (PAF) and/or lyso-phosphatidylcholine (lyso-PC), phosphatidylcholine (PC), and sphingomyelin (SM) were found in both the normal and dry eye rabbit tears. However, the number of types and the concentrations of SM molecules were markedly greater in the dry eye tears than in the normal tears. In addition, phosphatidylserine (PS) species that were readily detectable in dry eye tears were not found in normal tears. CONCLUSIONS: The combination of immobilized metal ion affinity chromatography and the solid ionic crystal matrix for MALDI enabled the detection and study of phosphorylated lipids in the tears. Specific differences between phospholipid levels in normal and dry eye tears were observable with this methodology. The appearance of various SM species only in the dry eye tears may provide markers for this disease state in the future.

Animals↗
PubMed1988

HSV-specific IgA from tears blocks virus attachment to the cell membrane.

The results of this investigation demonstrate that the IgA antibody in tears inhibits the attachment of herpes simplex virus (HSV) to cells and implicate immunoglobulin A (IgA) isotype antibody in the protection of the ocular surface. Tears were collected from the eyes of infected and uninfected rabbits and tested for the presence of infectious virus, for viral neutralization titer, and in an enzyme-linked immunoassay (ELISA) for antibody to HSV. Neither the tears from the uninfected rabbits nor the tears from the infected rabbits produced viral cytopathic effects on indicator cells in vitro. Tears from uninfected rabbits had no virus neutralization capacity while the tears from infected rabbits had titers of 1:10 to 1:40. Tears collected from HSV-1 infected rabbits, but not uninfected rabbits, had anti-HSV antibody titers as determined by the enzyme-linked immunosorbent assay. Lectin affinity column enrichment of IgA from the tears of uninfected and infected rabbits was examined for the capacity to block viral adsorption to cells using 32P-labeled virus. It was found that exposure of the radio-actively-labeled virus to the purified IgA fraction of tears from infected rabbits reduced the amount of virus binding to Vero cells, whereas the purified IgA obtained from uninfected tears had minimal activity. These results may imply a role for the IgA-containing fraction of tears from HSV-infected animals in preventing viral attachment to cells and suggest that antibody of this isotype plays a protective role at the ocular surface.

Animals↗
PubMed2001

[The influence of the hormonal replacement therapy on the amount and stability of the tear film among peri- and postmenopausal women].

PURPOSE: To estimate the amount of the tear-film and its stability among women treated due to endocrinological or gynecological indications with hormone replacement therapy. MATERIAL AND METHODS: 60 women treated with hormonal replacement therapy (HRT) were divided into 3 groups of examination: group 1 (20 women), treated with HRT after up to 1 year of menopausal symptoms, group 2 (20 women) treated with HRT between 1-5 years of menopausal symptoms, group 3 (20 women) treated with HRT after more than 5 years of menopausal symptoms. The secretion of the tear-film was measured using Schirmer I test and lisozyme test. The stability of the tear-film was measured with respect to the break-up time (BUT). The examinations were performed 3, 6, 12 months after HRT treatment. The values obtained before HRT treatment served as control. RESULTS: Before HRT treatment we observed significantly decreased tear secretion and lower stability in group 2 and group 3 as compared with group 1. We noticed the negative correlation between the secretion of the tear film as compared to age of examined women. HRT made the tear secretion significantly increased in group 2 and group 3 after 6, 12 and 3, 6, 12 months of HRT treatment respectively. We assessed a significant improvement of the tear film stability with respect to BUT in group 2 and 3 in all months of HRT treatment. We found significant correlation between modified Schirmer I test and lisozyme test (p < 0.001). CONCLUSION: The age and changes of the hormonal homeostasis have negative influence on the tear secretion, tear-film stability among peri and postmenopausal women. HRT treatment has beneficial influence on the tear film secretion and tear stabilization. Our results suggest that hormonal replacement therapy can be suitable for the treatment of the postmenopausal dry eye syndrome.

Aged↗