Aniko Rentka

Membrane array and multiplex bead analysis of tear cytokines in systemic sclerosis

Although serious ocular manifestations of systemic sclerosis (SSc) have been described, tear analysis of patients with SSc has not been performed in previous studies. Our aim was to measure a wide panel of cytokines and chemokines in tears of patients with SSc and to assess the most significant molecules with a more sensitive and specific method. Unstimulated tear samples were collected from nine patients with SSc and 12 age- and gender-matched healthy controls. The relative levels of 102 different cytokines were determined by a cytokine array, and then absolute levels of four key cytokines were determined by a magnetic bead assay. Array results revealed shifted cytokine profile characterized by predominance of inflammatory mediators. Of the 102 analyzed molecules, nine were significantly increased in tears of patients with SSc. Based on the multiplex bead results, C-reactive protein, interferon-γ-inducible protein-10, and monocyte chemoattractant protein-1 levels were significantly higher in tears of patients with SSc. Our current data depict a group of inflammatory mediators, which play a significant role in ocular pathology of SSc; furthermore, they might function as excellent candidates for future therapeutic targets in SSc patients with ocular manifestations.

Evaluation of commonly used tear sampling methods and their relevance in subsequent biochemical analysis

The human precorneal tear film is a special body fluid, since it is a complex mixture of proteins, lipids, small bioactive molecules, and their concentrations and relative distribution represent not only the metabolic state of the ocular surface but also the systemic and local homeostasis of the outer eye and the human body. This suggests that biochemical analysis of the precorneal tear film composition may provide a non-invasive tool for diagnosis and monitoring of disease progression or treatment efficacy in human medicine. However, collecting tears is demanding, and obtaining reproducible and unaltered samples is challenging because of the small sample volumes of tears. Several methods are available for tear collection as a preparatory step of precorneal tear film analysis, and the collection method used has to be assessed since it has a critical impact on the effectiveness of the assays and on the quality of the results. Each sampling method has advantages and disadvantages; therefore, it is not easy to choose the appropriate collecting method for tear collection. To overcome these limitations various methods have been recommended by different authors for special aspects of specific tests. The aim of our review was to evaluate tear sampling methods with regard to our ongoing biochemical analysis.

Vascular Endothelial Growth Factor in Tear Samples of Patients with Systemic Sclerosis.

Background. Systemic sclerosis is an autoimmune disease, characterized by widespread small vessel vasculopathy, immune dysregulation with production of autoantibodies, and progressive fibrosis. Changes in levels of proangiogenic cytokines had already been determined largely in serum. Our aim was to assess the levels of VEGF in human tears of patients with SSC. Patients and methods. Forty-three patients (40 female and 3 men, mean (SD) age 61 (48–74) years) with SSc and 27 healthy controls were enrolled in this study. Basal tear sample collection and tear velocity investigations were carried out followed by an ophthalmological examination. Total protein concentrations and VEGF levels were determined in tear samples. Results. The average collected tear fluid volume developed 10.4 μL (1.6–31.2) in patients and 15.63 μL (3.68–34.5) in control subjects. The average total protein level was 6.9 μg/μL (1.8–12.3) in tears of patients and control tears contained an average of 4.132 μg/μL (0.1–14.1) protein. In patients with SSc the average concentration of VEGF was 4.9 pg/μL (3.5–8.1) and 6.15 pg/μL (3.84–12.3) in healthy samples. Conclusions. Total protein production was increased because of the smaller tear volume. Decreased VEGF in tear of SSc patients can be explained also by the decreased tear secretion of patients.

Factors to be Considered when Performing Cataract Surgery in Patients Unable to Recline Flat

ABSTRACT Purpose: Although phacoemulsification is routinely performed with the patient in a supine position, certain locomotor deformities and other severe ailments can lead to the inability of the patient to recline flat. Methods: The present article surveys publications and summarizes our own experience regarding positioning techniques during cataract surgery on patients unable to adopt a supine position. Results: The successful and effective performance of cataract surgery by phacoemulsification depend on the comfortable positioning of both the patient and the surgeon. In order to achieve the highest possible success rate in phacoemulsification surgery, optimal positioning of the patient and the operating microscope are essential. There are other features of pivotal importance, including the construction of the operating table, the anesthesia technique, and the surgical approach. Conclusions: Surgeons often have to rely on their own imagination and experience in finding optimal positioning in order to perform effective and successful phacoemulsification surgery.

Corneal Manifestations of Systemic Sclerosis

ABSTRACT Purpose: Corneal involvement in systemic sclerosis (SSc) is rare, but due to rich collagen composition cornea is especially vulnerable to connective tissue diseases. Therefore, our aim was to evaluate corneal parameters of SSc patients. Methods: The study included 32 SSc patients and 39 control subjects with no ocular symptoms or ocular surface disorders. All study participants underwent Pentacam evaluation and objective signs of dry eye disease (DED), and clinical parameters were evaluated. Results: All pachymetric values, most of the corneal front surface, corneal volume, as well as anterior chamber depth measurements were significantly lower in the SSc group than in the control group (p < 0.05). Significant negative correlation was found between corneal parameters and age on the one hand, and disease duration on the other. Conclusions: Early recognition of corneal impairment, a possible extraintestinal manifestation of SSc, should be included in the check-up of the disease in order to reduce sight-threatening complications.

AB0632 Assessment of vegf and other cytokines in the tear of patients with systemic sclerosis

Background Systemic sclerosis (SSc) is an autoimmune disease, characterized by widespread small vessel vasculopathy, immune dysregulation with production of autoantibodies, and progressive fibrosis. SSc may be associated with sicca syndrome. Changes in levels of proangiogenic and proinflammatory cytokines had already been determined largely in serum, however, the local inflammatory and cytokine milieu in the tear of SSc patients has not yet been evaluated. Objectives We wished to determine VEGF and other cytokine and chemokine levels in tear samples of SSc patients. Methods First, forty-three patients (40 female and 3 men, mean (SD) age 61 (48–74) years) with SSc and 27 healthy controls were enrolled in the VEGF study. Basal tear sample collection and tear velocity investigations were carried out followed by an ophthalmological examination. Total protein concentrations and VEGF levels were determined in tear samples. In the multiple cytokine study, unstimulated tear samples were collected from nine patients with SSc and 12 age- and gender-matched healthy controls. The relative levels of 102 different cytokines were determined by a cytokine array, and then absolute levels of four key cytokines were determined by a magnetic bead assay. Results In the first study, the mean collected tear fluid volume developed 10.4 L (1.6–31.2) in patients and 15.63 L (3.68–34.5) in control subjects. The mean total protein level was 6.9 g/L (1.8–12.3) and 4.1 g/L (0.1–14.1) in tear samples of SSc patients and controls, respectively. In patients with SSc, the mean VEGF tear concentration was 4.9 pg/L (3.5–8.1) compared to 6.15 pg/L (3.84–12.3) in healthy samples. Multicytokine-array studies revealed shifted cytokine profile characterized by predominance of proinflammatory mediators in the tear samples of SSc patients. Out of the 102 analyzed proteins, nine were significantly increased in tears of patients with SSc. Based on the multiplex bead results, CRP, interferon-inducible protein 10 (IP-10) and monocyte chemoattractant protein-1 (mcp-1) levels were significantly higher in tears of patients with SSc compared to controls. Conclusions Impaired angiogenesis has been found by other investigators in SSc. This is reflected by lower VEGF levels in the tear samples of SSc patients compared to controls. The multi-cytokine array study revealed increased production of CRP and two important pro-inflammatory chemokines in the tear of SSc patients. Our current data depict a group of inflammatory mediators, which may play a significant role in ocular pathology of SSc. Disclosure of Interest None declared