The effect of artificial tear administration on visual field testing in patients with glaucoma and dry eye.

AIM: To examine the effects of artificial tear administration on perimetry of primary open-angle glaucoma patients with dry eye. METHODS: A total of 40 patients with primary open-angle glaucoma experienced in automated perimetry with symptoms of dry eye were enrolled in this study. After their pretest visit, they were instructed to use artificial tear four times a day in both eyes for 1 week. After 1 week, patients had visual field testing. Test taking time, reliability parameters (false-positive and false-negative errors) visual field indices and number of depressed points at different probability levels (P<5%, P<2%, P<1%, P<0.5%) in both total and pattern deviation plots were compared using paired Ttest. RESULTS: We found significant improvement in reliability parameters (false-positive errors from 2.4+/-2.1 to 2.1+/-1.9, P=0.02; and false-negative errors from 7.3+/-6.4 to 4.8+/-3.6, P=0.01) and visual field indices (MD increased from 5.97+/-5.61 to 4.57+/-4.53, P=0.001; PSD from 4.67+/-2.95 to 4.13+/-2.77, P=0.04 and SF decreased from 2.24+/-1.23 to 1.83+/-0.77, P=0.04) in the second testing after artificial tear administration. Test time significantly increased from 11.66+/-2.55 min to 14.26+/-1.36, P=0.001. The number of depressed points at probability levels P<1% (P=0.03) and P<0.5% (P=0.04) at total deviation plot and P<2% (P=0.02) and P<0.5% (P=0.009) in pattern deviation plot decreased significantly. CONCLUSION: Artificial tear administration in glaucomatous patients with dry eye seems to improve significantly reliability parameters and visual field indices.

Procollagen type I N-terminal propeptide (PINP) as an indicator of type I collagen metabolism: ELISA development, reference interval, and hypovitaminosis D induced hyperparathyroidism.

A sandwich enzyme-linked immunosorbent assay (ELISA) for quantification of the N-terminal propeptide of human procollagen type I (PINP) utilizing purified alpha 1-chain specific rabbit antibodies is described. The ELISA measured the content of the alpha 1-chain of PINP independent of the molecular form of the molecule. A parallelism was found between amniotic fluid (calibrator), normal and patient serum, and purified PINP (alpha 1), as well as the high and low molecular weight forms of PINP (alpha 1). The concentration of PINP in the calibrator (second trimester amniotic fluid) was determined to 25 micrograms/mL and the detection limit was 62 pg/mL measured in amniotic fluid, and 41 pg/mL measured in serum. The interassay coefficients of variation were 4.6% (low control) and 5.3% (high control), and the corresponding intraassay parameters were 2.9% and 4.9%. Recovery studies revealed an accuracy between 93% and 105%. The normal range (n = 57) for PINP was 56 ng/mL (median) the 10th and 90th centiles being 30 and 82 ng/mL, respectively. Patients with hyperparathyroidism due to hypovitaminosis D had median serum level of 168 ng/mL with a 10th centile of 44 ng/mL and a 90th centile of 450 ng/mL, these values being significantly different from the normal range (p < 0.001). The PINP-ELISA was superior to commercially available assays for PICP and osteocalcin in separation between healthy controls and patients with osteomalaci.

Aminopropeptide of human procollagen type I: a marker for the identification of blood from children in the mosquito blood meal.

A capture enzyme-linked immunosorbent assay (ELISA) to distinguish between blood from children and adults in the mosquito blood meal was examined using the alpha 1 chain of the aminopropeptide of human procollagen type I (PINP) as antigenic marker. Rabbit anti-human PINP (alpha 1) antibody was used as catching antibody, and either normal serum from 288 African and 58 Caucasian children and adults, or blood meals from 93 fed Aedes aegypti, were examined. PINP in excess of 40 optical density units (ODU) was detected in all children aged 0-13 years, whereas adults aged 21-77 years had PINP levels less than 25 ODU. In the transitional age group (14-20 years), the PINP levels ranged from 1 to 166 ODU. The PINP levels in 95% of the mosquito blood meals obtained from children exceeded the control levels, using 13 ODU as a cut-off value, whereas none of the blood meals from adults exceeded 13 ODU. The PINP levels in the mosquito blood meals were constant 1 and 8 h after ingestion, but they had decreased significantly after 16-19 h. Our data suggest that the test can be used to identify host preferences in studies of mosquitoes collected within 16 h after the blood meal. A field evaluation is necessary to determine the potential of the antigenic marker PINP as a tool in the identification of mosquito host preference.

The effect of cholesterol on the morphology and reactivity of the mixture of lipids used in syphilis serology.

The effect of increasing amounts of cholesterol on the morphology of the liposomes constituting the VDRL-antigen was studied. The morphological parameters examined were the shape of the lipoidal particles and especially the number of lamellae on each particle in the various mixtures of lipids studied. Cholesterol in the presence of cardiolipin and lecithin is observed as rhomboid crystals, indicating that the majority of the cholesterol is located exterior to the lamellar membranes of lecithin and cardiolipin. It is shown that the effect of cholesterol is to reduce the number of individual lamellae per liposome, mainly by mechanically dispersing the cardiolipin and lecithin on the surface of the cholesterol crystals. It is suggested that cholesterol has no effect on the structure of the epitopes which react with antibodies in sera from patients with syphilis, but that, as a result of the mechanical dispersion of cardiolipin and lecithin, it creates liposomes with more accessible epitopes.

The specificity of anti-cardiolipin antibodies from syphilis patients and from patients with systemic lupus erythematosus.

In order to elucidate the fine specificity of anti-cardiolipin antibodies (ACA) in patients with SLE compared to patients with syphilis (SY) various inhibition experiments were performed. Seven SLE sera and eight SY sera positive for ACA were diluted and preincubated with either cardiolipin VDRL-antigen, mitochondial particles, dsDNA, ssDNA or dilution buffer. The sera were subsequently assayed for residual ACA activity of IgG or IgM class using a sensitive ELISA technique. Significant inhibition of IgM ACA activity in SLE sera was found with cardiolipin, VDRL-antigen and mitochondrial particles. Cardiolipin inhibited binding to a significantly higher extent than the other antigens. In SY sera significant inhibition of the IgM ACA activity was found with all antigens used. The strongest inhibition was seen using VDRL-antigen. Inhibition of IgG ACA activity could only be clearly estimated in SY sera where VDRL-antigen was found to be a much stronger inhibitor than the rest, purified cardiolipin being the weakest. Only two out of seven SLE sera were IgG ACA positive which made a clear conclusion impossible but a strong inhibitory capability of pure cardiolipin and a weaker inhibition with VDRL-antigen was found. This study disclosed a difference between SLE and SY sera showing strong reactivity of ACA in SLE sera with purified cardiolipin, contrasting to ACA in SY sera which predominantly reacted with cardiolipin in the liposome environment, as found in the VDRL-antigen and in mitochondrial particles.

Enzyme-linked immunosorbent assay for detection of antibodies to the venereal disease research laboratory (VDRL) antigen in syphilis.

An enzyme-linked immunosorbent assay (ELISA) for detection of immunoglobulin G (IgG) and IgM to cardiolipin, lecithin, and cholesterol (VDRL [Venereal Disease Research Laboratory] ELISA) is described. The specificity of the VDRL ELISA for IgG and IgM was 99.6 and 99.5%, respectively, with sera from 1,008 persons without syphilis. For a group of patients with false-positive results in traditional nontreponemal tests and for patients with autoimmune diseases, the VDRL ELISA for IgG had a higher specificity than the VDRL ELISA for IgM. The sensitivity for IgG and IgM with 118 sera from patients with untreated syphilis was 96.6 and 94.9%, respectively, which was equivalent to the sensitivities of the traditional nontreponemal tests. The performance of the VDRL ELISA was compared with that of an ELISA that uses cardiolipin as the antigen (cardiolipin ELISA). The VDRL ELISA was significantly more sensitive (P less than or equal to 0.01) than the cardiolipin ELISA with 25 sera from syphilis patients but was less sensitive (P less than or equal to 0.01) with 53 sera from patients with autoimmune diseases. The antibody reactivity in the VDRL ELISA could not be absorbed out by lecithin and cholesterol, and the sera from patients with syphilis did not react in an ELISA that uses cholesterol and lecithin as the antigen. This indicates that cholesterol and lecithin, although not antigenic by themselves, may change the structural form of the epitope on cardiolipin so that it becomes more recognizable for antibodies in syphilis and less recognizable for antibodies in autoimmune diseases. The results of the VDRL ELISA were expressed in percentages of the absorbance value of a positive control. The VDRL ELISA gave, without titration of sera, quantitative results that correlated with the quantitative results of the traditional nontreponemal tests obtained by titration. The VDRL ELISA will be well suited for large-scale testing for syphilis and may replace other nontreponemal tests.