Daniela Suesskind

Keratoepithelin in secondary corneal amyloidosis

BackgroundAmyloid is found in several corneal dystrophies, including distinct lattice corneal dystrophies (LCD) and Avellino corneal dystrophy. Recently, point mutations in the transforming growth factor-beta-induced gene (TGFBI) encoding for keratoepithelin (KE) have been demonstrated in these corneal disease entities. We intended to investigate if KE was also a component of the rarely seen secondary corneal amyloid deposits.MethodsImmunohistochemical staining with a polyclonal antibody against KE was performed on formalin-fixed paraffin-embedded tissue of five corneal buttons with secondary amyloid obtained after keratoplasty. Secondary amyloidosis was due to Fuchs´ endothelial dystrophy (FED) with bullous keratopathy and/or recurrent erosions in all cases. The diagnosis had been established by light microscopy using Congo red staining. Two cases of LCD type I served as positive controls and three corneas with FED and one with keratoconus without amyloid served as negative controls.ResultsAll corneas with secondary amyloidosis as well as LCD type I revealed positive staining in the respective amyloid deposits. KE was localized in the subepithelial pannus and in the anterior stroma in the corneas with secondary amyloidosis. In the specimens with LCD type I it was distributed in the amyloid deposits located in the anterior and mid-stroma. Staining for KE showed a granular appearance in all cases. The intensity of staining was variable among the specimens.ConclusionsKE is found not only in primary amyloid deposits of hereditary corneal dystrophies, but also in secondary amyloidosis of the cornea of diverse ethiologies.

Circulating melanoma cells in peripheral blood of patients with uveal melanoma before and after different therapies and association with prognostic parameters: a pilot study

Acta Ophthalmol. 2011: 89: 17–24

GDF-15: a novel serum marker for metastases in uveal melanoma patients

BackgroundAbout 50% of patients with uveal melanoma (UM) develop metastases during the course of their disease. We analyzed serum levels of Growth Differentiation Factor-15 (GDF-15), with the aim of identifying patients with early metastases.MethodsGDF-15 concentration was measured using an enzyme-linked immunosorbent assay (ELISA) in serum samples from 188 UM patients (170 patients without metastases; 18 patients with clinically detectable metastases) and 18 healthy control individuals. Data were analyzed with respect to differences between patients with and without clinically detectable UM metastases. GDF-15 serum levels were further analyzed with regard to significant patient and tumor characteristics as revealed by histology and multiplex ligation-dependent probe amplification (MLPA) to determine chromosome 3 copy number. GDF-15 expression in UM was investigated by immunohistochemistry.ResultsPatients with clinically detectable metastases had significantly higher GDF-15 serum levels compared to those without clinically detectable metastases as well as to healthy individuals (ANOVA; pu2009<u20090.001). GDF-15 concentrations in UM patients with overt clinically detectable metastases were significantly higher than those in UM patients with a second malignancy in remission but without clinically detected UM metastases (ANOVA; pu2009<u20090.001). No association between serum concentration of GDF-15 and clinical, pathological, and genetic features was observed. GDF-15 protein was only expressed in a minority of UM cells in most tumors.ConclusionsOur data suggest that GDF-15 can be used as a serum marker for the diagnosis of metastases in UM patients. Further data collection and analysis are necessary to evaluate a possible prognostic role of GDF-15 in predicting early metastases.

Sulcus anatomy and diameter in pseudophakic eyes and correlation with biometric data: evaluation with a 50 MHz ultrasound biomicroscope.

PURPOSE: To evaluate the sulcus anatomy and possible correlations between sulcus diameter and white‐to‐white (WTW) diameter in pseudophakic eyes, data that may be important in the stability of add‐on intraocular lenses (IOLs). SETTING: University Eye Hospital, Tuebingen, Germany. DESIGN: Case series. METHODS: In pseudophakic eyes, the axial length (AL) and horizontal WTW were measured by the IOLMaster device. Cross‐sectional images were obtained with a 50 MHz ultrasound biomicroscope on the 4 meridians: vertical, horizontal (180 degrees), temporal oblique, and nasal oblique. Sulcus‐to‐sulcus (STS), angle‐to‐angle (ATA), and sclera‐to‐sclera (ScTSc) diameters were measured. The IOL optic diameter (6.0 mm) served as a control. To test reliability, optic measurements were repeated 5 times in a subset of eyes. RESULTS: The vertical ATA and STS diameters were statistically significantly larger than the horizontal diameter (P=.0328 and P=.0216, respectively). There was no statistically significant difference in ScTSc diameters. A weak correlation was found between WTW and horizontal ATA (r = 0.5766, P<.0001) and between WTW and horizontal STS (r = 0.5040, P=.0002). No correlation was found between WTW and horizontal ScTSc (r = 0.2217, P=.1217). CONCLUSIONS: The sulcus anatomy had a vertical oval shape with the vertical meridian being the largest, but it also had variation in the direction of the largest meridian. The WTW measurements showed a weak correlation with STS. In pseudophakic eyes, Soemmerring ring or a bulky haptic may affect the ciliary sulcus anatomy. Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned.

Pars plana vitrectomy for treatment of advanced Coats’ disease—presentation of a modified surgical technique and long-term follow-up

BackgroundTo present a modified surgical technique in the treatment of retinal detachment secondary to advanced Coats’ disease in children, and report on long-term anatomical and functional outcome.MethodsWe analysed an interventional case series of 13 patients (13 eyes) with advanced Coats’ disease characterised by retinal detachment in addition to massive subretinal exudates and vascular malformation. The presented patients underwent pars plana vitrectomy (PPV), including a modified technique of exocryotherapy applied after fluid–air exchange in order to achieve complete treatment of the vascular changes, to reduce associated side-effects, and to avoid retinectomy and silicone oil tamponade.ResultsWithin a median follow-up period of 37xa0months (range: 18–66xa0months), no enucleation was necessary. Four eyes (31xa0%) did not need any further therapy, and in nine eyes (69xa0%) additional treatments were performed. Six patients (46xa0%) required revisional surgery with silicone oil tamponade. In ten eyes (77xa0%), the pathologic vessels and exudates finally regressed and the retina reattached. Visual acuity (VA) could be stabilized in the majority of patients: in three eyes (27xa0%) VA improved, in four eyes (36xa0%) VA remained stable, in four eyes (36xa0%) visual acuity (VA) deteriorated, and in two eyes VA could not be evaluated.ConclusionsThe presented modified technique allows for sufficient cryotherapy of vascular malformations, even in the presence of massive exudation, in a subset of patients with advanced Coats’ disease, and thus may reduce surgery-related complications and improve the rehabilitation process of these young patients.

Conjunctival amyloidosis — clinical and histopathologic features

PurposeConjunctival amyloidosis is a rare disorder. It is often clinically not suspected or diagnosed. This study intended to demonstrate the clinical and histopathologic features of this infrequent disease, including an immunohistochemical search for amyloidotic proteins.MethodsRetrospective case series of the clinical and histopathologic characteristics of six patients with conjunctival amyloidosis. Immunohistochemical analysis with respect to possible amyloidotic components of the conjunctival deposits was performed.ResultsThe diagnosis of amyloidosis was not suspected in all six cases presenting with an amelanotic conjunctival lesion. In three patients a conjunctival tumor of unknown origin, in one case each a papillomatous alteration of the conjunctiva, a conjunctival granulomatous inflammation, and a lymphoma were assumed respectively. The diagnosis of amyloidosis was made by histopathology. Immunohistochemical examination found lambda and kappa light chains as well as prealbumin within the amyloid deposits in one of the six specimens.ConclusionsThe diagnosis of amyloidosis has to be kept in mind in cases with an unclear conjunctival mass or inflammatory process. Only a tissue biopsy is able to prove the diagnosis. A possible underlying systemic disease has to be ruled out.

CRITICAL EVALUATION OF THE USABILITY OF AUGMENTED REALITY OPHTHALMOSCOPY FOR THE TRAINING OF INEXPERIENCED EXAMINERS

Purpose: To measure the value of augmented reality technology usage to teach the medical students performing binocular indirect ophthalmoscopy. Methods: Thirty-seven medical students were randomly assigned to the training of binocular indirect ophthalmoscopy either in the conventional way or with augmented reality ophthalmoscopy (ARO). For testing students skills, they had to examine a real person using a conventional ophthalmoscopy system and draw the optic disk. They also had to fill out a questionnaire. Subjective and objective evaluations were performed. Results: Thirty-seven students were randomly assigned to two groups. Eighteen students were trained with conventional ophthalmoscopy and 19 students with ARO. The questionnaires showed no differences. Performing an objective analysis, the median ophthalmoscopy training score for the conventional ophthalmoscopy group was 1.2 (range, 0.67–2) and showed a significant difference (P < 0.0033) to the ARO group (median 2; range, 0.67–2). Conclusion: The study provides evidence that a single ARO training is efficient to improve ophthalmoscopy skills. As the objective analysis showed, the ARO group had a significantly superior performance. Our study also indicates that subjective evaluation of the fundus drawings without systematic analysis is prone to errors.

Cataract formation: a possible complication of intra-arterial chemotherapy for retinoblastoma.

Purpose To delineate and discuss a not yet described possible ocular complication of selective intra-arterial chemotherapy (SIAC) for treatment of retinoblastoma. Methods A 23-month-old girl with a large unilateral retinoblastoma was treated with repeated SIAC using 5 mg melphalan between July 2010 and January 2012. Clinical course of tumor and further ocular changes after therapy and histopathologic findings are described. Results In total, 5 SIAC were performed over a time period of 18 months. After the last SIAC, diffuse dense cataract prevented further funduscopy. In addition, anterior chamber seeding was obvious, leading to the decision to enucleate the eye. Histopathologically, nearly complete regression of the main tumor mass with prominent calcifications, but vital tumor seeding in the vitreous, on the lens surface, on the ciliary body, and in the anterior chamber, was observed. Peculiar vacuolation of the lens epithelial cells, liquefaction of the subepithelial lens fibers, and diffuse small vacuoles within the lens were striking. Conclusions Repeated SIAC with melphalan may induce cataract formation, possibly as a toxic effect of the chemotherapeutic to the lens, maybe combined with radiation exposure during fluoroscopy. This ocular complication should be taken into consideration as a limitation of the number of feasible repeated treatments.

Retrospective Evaluation of Patients With Uveal Melanoma Treated by Stereotactic Radiosurgery With and Without Tumor Resection

IMPORTANCEnThe present study intended to analyze the suitability of single-dose stereotactic radiotherapy in the treatment of uveal melanoma that cannot be handled with ruthenium-brachytherapy and therefore is a challenge for ophthalmologists concerning local tumor control, as well as preservation of the eye and visual function.nnnOBJECTIVESnTo evaluate local tumor control, eye preservation, visual course, radiation complications, metastases, and death after single-dose stereotactic radiotherapy (SDRT) applied exclusively or combined with tumor resection in uveal melanomas that are neither suitable nor favorably located for ruthenium brachytherapy.nnnDESIGNnRetrospective, observational case series.nnnSETTINGnPrimary care center.nnnPARTICIPANTSnSeventy-eight patients with uveal melanoma were treated.nnnINTERVENTIONnBetween June 3, 2003, and March 18, 2008, patients with uveal melanoma received SDRT monotherapy (group 1, 60 patients) or SDRT combined with tumor resection (group 2, 18 patients). Radiotherapy was performed with a tumor-surrounding dose of 25 Gy on a linear accelerator.nnnMAIN OUTCOME MEASURESnLocal tumor control, eye preservation, visual results, and radiation complications.nnnRESULTSnWithin a median follow-up of 33.7 months (range, 0.13-81.13 months), 6 recurrences occurred in group 1; none recurred in group 2. The Kaplan-Meier estimate for local control was 85% at 3 years in group 1 and 100% in group 2 (P = .22). Eye preservation rate was 77% vs 87% at 3 years (groups 1 and 2, respectively) (P = .82). Visual acuity decreased with a median loss of -18 Snellen lines (group 1) and -22 Snellen lines (group 2). More retinopathies (P = .07), opticopathies (P = .27), and rubeotic glaucomas (P = .10) occurred in group 1. No significant difference was observed in the development of metastases (P = .33). The groups differed in overall survival because of 2 deaths occurring shortly after surgery in group 2 for unexplained reasons (P = .06).nnnCONCLUSIONS AND RELEVANCEnSurvival analysis suggested that SDRT with combined tumor resection might be associated with increased tumor control and fewer radiation complications than SDRT as monotherapy. Both groups had similar eye retention rates and were comparable concerning the decrease in visual function in most eyes. However, the protocol was stopped after 3 unexplainable deaths after surgery.

Characterisation of novel uveal melanoma cell lines under serum-free conditions

BackgroundThe establishment of long-term uveal melanoma (UM) cell lines is difficult. However, studying living cells and their behaviour in the presence of other cells and the extracellular matrix is important in terms of understanding tumour biology and malignant behaviour. We have established three UM cell lines and report a first characterisation of these cell lines.MethodsThree established UM cell lines (UMT2, UMT26 and UMT33) were analysed according to their morphologic characteristics, melanocytic differentiation, adhesion on different extracellular matrices and proliferative activity. Copy number changes of chromosomes 1, 3, 6 and 8 were studied by multiplex ligation-dependent probe amplification (MLPA). Oncogenic mutations in UM involving exons 4 and 5 of GNAQ and GNA11, respectively, were analysed by sequencing.ResultsAll cell lines grew in suspension. UMT2 cells were homogeneous, UMT26 and UMT33 cells heterogeneous with regard to cell size and pigmentation. All UM cell lines revealed a melanocytic differentiation. UMT2 and 33 adhered on various extracellular matrices, while UMT26 only adhered to basal membrane extract (BME). This difference corresponded to the different expression of various integrins. Ki67 was expressed by 89xa0% of UMT2 and 95xa0% of UMT33 cells, which thus were in a proliferative stage, while only 2xa0% of UMT26 cells revealed immunostaining for this proliferation marker. The doubling time of UMT2 was 3xa0days, 12xa0days for UMT33, and circa 3–4xa0months for UMT26. MLPA revealed disomy 3 in UMT2 and monosomy 3 in UMT33. The same point mutation was found in UMT2, 26 and 33, in exon 5 of GNA11 at codon 209 (p.Q209L).ConclusionsThe establishment of UM cell lines under serum-free conditions is possible. Characterisation of UMT2, 26, and 33 revealed obvious differences in cytomorphology, melanocytic differentiation, adhesion on extracellular matrices, and proliferative activity. UMT2, 26 and 33 showed the same oncogenic mutation in exon 5 of GNA11.