Jan Ulrik Prause

Malignant melanoma of the conjunctiva: occurrence and survival following surgery and radiotherapy in a Danish population

Abstract• Purpose: To describe patient characteristics, risk of recurrence, and survival in patients with premalignant and malignant melanocytic lesions of the conjunctiva. • Methods: In a retrospective study, 55 cases with a minimum follow-up of 10 years were reviewed. • Results: The actuarial 5-and 10-year survival rates were 86% (95% Cl: 76–96%) and 73% (95% Cl: 60–87%) respectively. The 5-year risk of local recurrence was 35% (95% Cl: 21–49%), and the 10-year risk was 43% (95% Cl: 28–58%). Primary acquired melanosis with atypia was the first manifestation in 13 cases, malignant melanoma in 42 cases. No significant difference in survival and recurrence was found between the two groups or by gender, location of primary lesion, histological characteristics, or extent of surgical procedure. A minimum incidence of 0.052 cases a year per 100000 in Denmark was calculated. • Conclusion: The results are estimated to represent a nationwide sample of patients. No prognostic factors of significance were identified.

Primary lymphoma of the lacrimal sac: an EORTC ophthalmic oncology task force study

Aim: To define the clinical and histopathological characteristics of primary lacrimal sac lymphoma in a predominantly white population. Methods: Specimens of lacrimal sac lymphoma and follow up data were solicited from members of the Ophthalmic Oncology Task Force of the European Organization for Research and Treatment of Cancer (EORTC) and the European Ophthalmic Pathology Society (EOPS). Specimens were stained with haematoxylin and eosin and an immunohistochemical panel against leucocyte antigens was applied. Diagnosis was reached by consensus of five experienced pathologists according to the World Health Organization classification system. The histopathological findings were correlated with the clinical data. Results: Of 15 primary lacrimal sac lymphomas, five (33%) were diffuse large B cell lymphoma (DLBCL), five (33%) were extranodal marginal zone B cell lymphoma of mucosa associated lymphoid tissue (MALT lymphoma), three were classified as “transitional MALT lymphoma,” being in transition from MALT lymphoma to DLBCL, and two were unclassified B cell lymphomas. Nine of the patients were female, and the median age at the time of diagnosis was 71 years (range 45–95 years). The most frequent presenting symptoms were epiphora (85%), swelling in the region of the lacrimal sac (79%), and dacryocystitis (21%). All but one patient presented in stage I. Systemic spread occurred in three of nine patients (33%). The 5 year overall survival was 65%. Conclusions: DLBCL and MALT lymphoma are equally common in the lacrimal sac in contrast with the remaining periorbital and/or orbital region where MALT lymphoma predominates.

Cotransport of H+, lactate, and H2O in porcine retinal pigment epithelial cells.

The retinal pigment epithelium (RPE) of the eye transports water and lactate ions in the direction from retina to choroid. The water transport is important in maintenance of retinal adhesion and the transport of lactate ions serves to regulate the lactate levels and pH of the subretinal space. This study investigates by means of a non-invasive technique the mechanism of coupling between transport of H(+), lactate ion, and water in the monocarboxylate transporter (MCT1) located in the apical (retinal) membrane of a mammalian RPE. Primary cultures of porcine RPE cells were grown to confluence and placed in a perfusion chamber in which the solution facing the retinal membrane could be changed rapidly. Two types of experiments were performed: Changes in cell water volume were measured by self-quenching of the fluorescent dye Calcein, and changes in intracellular pH were measured ratiometrically using the fluorescent dye BCECF. In lactate-free solutions, mannitol addition to the retinal bath caused intracellular acidification and cell shrinkage, given by a single osmotic water permeability of 1.2+/-0.1 x 10(-4)cmsec(-1) (osmoll(-1))(-1). In solutions containing 50 mmoll(-1) lactate, however, the mannitol-induced cell shrinkage was faster and the cells alkalinized. These effects were not linear functions of the magnitude of the imposed osmotic gradients: Both volume effects and changes in intracellular pH showed apparent saturation with increasing gradients. Abrupt isosmotic replacement of Cl(-) with lactate in the concentration range from 3 to 50 mmoll(-1) caused an immediate cell swelling as well as an immediate intracellular acidification; both effects showed apparent saturation with increasing lactate concentration. The K(m) values were: 11+/-2 mmoll(-1) for the water fluxes and 13+/-4 mmoll(-1) for the H(+) and lactate fluxes. The data suggest that H(2)O is cotransported along with H(+) and lactate ions in MCT1 localized to the retinal membrane. The study emphasizes the importance of this cotransporter in the maintenance of water homeostasis and pH in the subretinal space of a mammalian tissue and supports our previous study performed by an invasive technique in an amphibian tissue.

Relation between blink frequency and break-up time?

Abstract. A possible correlation between break‐up time (BUT) and blink frequency was investigated in 32 healthy persons (17 females and 15 males) and in 29 patients suffering from primary Sjögrens syndrome (27 females and 2 males). A significant negative correlation, was found between BUT and blink frequency. In persons with high BUT, the blink frequency is low, probably just sufficient to prevent spontaneous tear breaks. In persons with low BUT, i.e. the Sjögren patients, the blink frequency is high, and tear fluid breaks are mostly prevented.

Human papillomavirus in normal conjunctival tissue and in conjunctival papilloma. Types and frequencies in a large series.

Aim: To examine conjunctival papilloma and normal conjunctival tissue for the presence of human papillomavirus (HPV). Methods: Archival paraffin wax-embedded tissue from 165 conjunctival papillomas and from 20 histological normal conjunctival biopsy specimens was analysed for the presence of HPV by PCR. Specimens considered HPV positive using consensus primers, but with a negative or uncertain PCR result using type-specific HPV probes, were analysed with DNA sequencing. Results: HPV was present in 86 of 106 (81%) β-globin-positive papillomas. HPV type 6 was positive in 80 cases, HPV type 11 was identified in 5 cases and HPV type 45 was present in a single papilloma. All the 20 normal conjunctival biopsy specimens were β-globin positive and HPV negative. Conclusion: There is a strong association between HPV and conjunctival papilloma. The study presents the largest material of conjunctival papilloma investigated for HPV and the first investigation of HPV in normal conjunctival tissue. HPV types 6 and 11 are the most common HPV types in conjunctival papilloma. This also is the first report of HPV type 45 in conjunctival papilloma.

Increasing incidence of ophthalmic lymphoma in Denmark from 1980 to 2005

PURPOSE To evaluate patient characteristics and incidence of ophthalmic lymphoma in Denmark during the period 1980 to 2005. METHODS All patients in Denmark with a diagnosis of ophthalmic lymphoma during the period 1980 to 2005 were retrieved from three different population-based registries. Specimens from all patients were collected and reclassified according to the World Health Organization (WHO) classification system. Incidence rates were calculated by using Poisson regression models. RESULTS A total of 228 patients with a histologically verified diagnosis of ophthalmic lymphoma were included. There was an equal distribution of males and females. The most frequent lymphoma subtype was extranodal marginal zone B-cell lymphoma (MALT [mucosa-associated lymphoid tissue] lymphoma, 55.5%) and most cases were located in the orbit (56.8%). High-grade lymphoma subtypes were found more frequently in males than in females. Incidence rates were highly dependent on the patients age. For all ages, a statistically significant annual average increase of 3.4% during the 26-year period was found. This increase was primarily due to a rise in the incidence of MALT lymphoma. CONCLUSIONS In the Danish population ophthalmic lymphoma consists primarily of orbital MALT lymphoma. Although it is a rare disease in mostly elderly patients, the incidence of ophthalmic lymphoma is increasing at a rapid pace.

Mantle cell lymphoma in the orbital and adnexal region

Aims: To characterise clinicopathological features of mantle cell lymphoma (MCL) in the orbital and adnexal region. Methods: Data on lymphoid lesions were retrieved searching the Danish Ocular Lymphoma Database 1980–2005. Specimens were collected from Danish pathological departments and re-evaluated with a panel of monoclonal antibodies. For all patients with confirmed MCL the complete clinical files were collected and reviewed. Results: Twenty-one patients with MCL in the orbital and adnexal region were identified comprising 9% (21/230) of all lymphoma in the ocular region. There were 18 male patients and three female patients with an age range from 60 to 90 years (median 75 years). Orbital and adnexal region MCL as first presenting symptom comprised 67% of the patients. Of these, 71% had bilateral involvement. The orbit (71%) and eyelids (64%) were the most commonly affected sites. All but two presented in stage III/IV. Secondary MCL comprised 33% of the patients. Bilateral affection (29%) was less common in this patient group. The median survival was not different between the two presentation groups. Patients receiving anti-CD20 (rituximab)-containing chemotherapy had a significantly better 5-year overall survival (OS) rate (83%) than patients in treatment regimes without rituximab (5-year OS rate, 8%). Conclusions: Orbital and adnexal region MCL presents in elderly males. The orbit and eyelid are frequently involved. There is a very high proportion of systemic involvement in general with MCL of the orbital and adnexal region. Most patients presented with stage IV disease and had multiple relapses and short survival time. Treatment with rituximab-containing chemotherapy improved survival significantly compared with combination chemotherapy without rituximab.

Collection and concentration of tear proteins studied by SDS gel electrophoresis Presentation of a new method with special reference to dry eye patients

An obvious obstacle in the analysis of tear film samples from patients with dry eyes is the drastically reduced tear volume. To overcome this problem we have developed a method by which the surface of the eye is flushed with saline, diluting the sparse tear fluid, followed by a concentration of the wash fluid. We compared undiluted tears, diluted tears, tear fluid collected by flushing, reconcentrated diluted tears and reconcentrated flush fluid. Gel electrophoresis of the tear samples obtained showed a representative collection of tear proteins, with bands ranging in molecular weight from about 10 kDa to about 90 kDa. The method may therefore provide a useful alternative to the analysis of undiluted tear fluid in such patients. After storage at −80° C for 1 month, electrophoresis often showed some weakening of band intensities. However, no specific loss of bands was found, and by ultracentrifugation a band profile similar to that of fresh, undiluted tears was obtained. For most purposes, therefore, samples may be kept by this means for later analysis.

Ocular findings in cystic fibrosis patients receiving vitamin A supplementation.

Abstract• Background: Vitamin A deficiency with eye symptoms has been reported in patients with cystic fibrosis who received the recommended daily intake of vitamin A. • Methods: We measured serum retinol, dark adaptation, contrast sensitivity, and dry eye status in 35 adult cystic fibrosis patients to ascertain whether they had ocular signs or symptons. • Results: Median serum retinol concentration was 1.95 μmol/l, range 1.08–4.01 μmol/l, with no values indicating vitamin A deficiency. Retinal light sensitivity was normal. Nineteen patients had reduced contrast sensitivity. Conjunctival imprints all showed plenty of goblet cells, but were characteristic of dry eye in 42% of patients (n=14). Decreased tear film stability was found in 49% (n=17), tear production was low in 31% (n=11), and 23% (n=8) showed an increased amount of dying epithelial cells. Nine patients (26%) had keratocon-junctivitis sicca according to the Copenhagen criteria. • Conclusion: Our patients had no biochemical or clinical signs of vitamin A deficiency. We speculate that the high incidence of dry eye could be a primary manifestation of cystic fibrosis.

Cell proliferation activity in posterior uveal melanoma after Ru-106 brachytherapy: an EORTC ocular oncology group study

AIM To evaluate the cell proliferation activity in posterior uveal melanomas after Ru-106 brachytherapy. METHODS Eyes containing choroidal or ciliary body melanoma from seven ocular oncology centres, which were enucleated after first being treated by Ru-106 brachytherapy and which had enough melanoma tissue to enable histological assessment, were included. The 57 eligible specimens were divided into a group of 44 eyes that were enucleated because of tumour regrowth, and a non-recurrent group of 13 eyes that were enucleated because of complications such as neovascular glaucoma. 46 non-irradiated eyes harbouring uveal melanoma served as a control group. All specimens underwent routine processing. They were cut into 5 μm sections, and were stained with two main cell proliferation markers: PC-10 for PCNA and MIB-1 for Ki-67. The stained sections were assessed, and the cells that were positive in the immunostaining were counted in each section. The results were evaluated by various statistical methods. RESULTS The PC-10 score showed a statistically significant difference across the three groups (p = 0.002). The control group showed the highest PC-10 score (median 31.0 PCC/HPF) followed by the tumour regrowth group (median 4.9 PCC/HPF). The lowest PC-10 scores were found in the non-recurrent tumours (median 0.05 PCC/HPF). The MIB-1 score in the control group (median 5.77 PCC/HPF) was similar to the regrowth group (median 5.4 PCC/HPF). In contrast, the MIB-1 score in the non-recurrent tumours was statistically significantly lower (median 0.42 PCC/HPF). The PC-10 and MIB-1 scores were similar in tumours composed of either spindle cells or epithelioid cells in all groups. CONCLUSIONS The non-recurrent melanomas demonstrate significantly lower cellular proliferation activity than melanomas that showed regrowth or that were not irradiated at all. In our hands, PCNA gave more meaningful information than Ki-67. Our findings strongly support the need for treating regrowing posterior uveal melanoma either by enucleation or re-treatment by brachytherapy. On the other hand, also in the non-recurrent uveal melanomas there are viable cells with potential for proliferation, although fewer in number, with unknown capacity for metastatic spread. Therefore, the irradiated tumours should be followed for many years, probably for life.