A practical approach to estimating optic disc dose and macula dose without treatment planning in ocular brachytherapy using 125I COMS plaques.

BACKGROUND: It has been reported that proximity of the tumor to the optic disc and macula, and radiation dose to the critical structures are substantial risk factors for vision loss following plaque brachytherapy. However, there is little dosimetry data published on this. In this study, therefore, the relationship between distance from tumor margin and radiation dose to the optic disc and macula in ocular brachytherapy using 125I Collaborative Ocular Melanoma Study (COMS) plaques was comprehensively investigated. From the information, this study aimed to allow for estimation of optic disc dose and macula dose without treatment planning. METHODS: An in-house brachytherapy dose calculation program utilizing the American Association of Physicists in Medicine Task Group-43 U1 formalism with a line source approximation in a homogenous water phantom was developed and validated against three commercial treatment planning systems (TPS). Then optic disc dose and macula dose were calculated as a function of distance from tumor margin for various tumor basal dimensions for seven COMS plaques (from 10 mm to 22 mm in 2 mm increments) loaded with commercially available 125I seeds models (IAI-125A, 2301 and I25.S16). A prescribed dose of 85 Gy for an irradiation time of 168 h was normalized to a central-axis depth of 5 mm. Dose conversion factors for each seed model were obtained by taking ratios of total reference air kerma per seed at various prescription depths (from 1 mm to 10 mm in 1 mm intervals) to that at 5 mm. RESULTS: The in-house program demonstrated relatively similar accuracy to commercial TPS. Optic disc dose and macula dose decreased as distance from tumor margin and tumor basal dimension increased. Dose conversion factors increased with increasing prescription depth. There existed dose variations (<8%) among three 125I seed models. Optic disc dose and macula dose for each COMS plaque and for each seed model are presented in a figure format. Dose conversion factors for each seed model are presented in a tabular format. CONCLUSIONS: The data provided in this study would enable clinicians in any clinic using 125I COMS plaques to estimate optic disc dose and macula dose without dose calculations.

Assessment of p16 expression and HPV infection in adenoid cystic carcinoma of the lacrimal gland.

Purpose: Adenoid cystic carcinoma (ACC) in the lacrimal gland is a rare malignancy. P16 is encoded by the CDKN2A gene, which is recognized as a tumor suppressor due to its inactivation in many types of tumors. However, p16 overexpression is also linked to adverse tumor parameters. These contradictory observations have also been confirmed in ACCs in the salivary glands. Furthermore, evidence of human papilloma virus (HPV) infection is found in a proportion of ACCs in the salivary glands. P16 is often overexpressed in HPV-related squamous cell carcinoma in parallel. To our knowledge, the role of p16 and HPV in ACCs in the lacrimal gland is still unknown. Methods: Twenty-one ACCs in the lacrimal gland and ten matched healthy lacrimal glands were studied. P16 was detected with immunohistochemistry (IHC), and HPV was detected with in situ hybridization (ISH) and PCR in all cases. Other cell cycle proteins were also detected with IHC, including cyclin D1 and Ki67. The methylation status of the p16 promoter was detected with methylation-specific PCR (MSP) to further investigate the regulation of p16 expression. Results: The expression rates of p16 (47.6%, 10/21), cyclin D1 (100%, 21/21), and Ki67 (52.4%, 11/21) were increased in ACCs compared to healthy lacrimal glands (negative). The results showed p16 expression was limited to the inner ductal epithelial cells in the majority of the tubular and cribriform patterns. In solid ACCs, p16 was uniformly positive. HPV was negative in all 21 cases with ISH and PCR. P16 overexpression was associated with cyclin D1 overexpression (p=0.013). Only 13 cases were tested successfully with MSP. The expression rate of p16 methylation was 23.1% (3/13) of the ACCs. Compared with primary ACCs, recurrent ACCs showed higher p16, cyclin D1, and Ki67 expression (p=0.011, p=0.026, p=0.049, respectively). Conclusions: In summary, p16 overexpression was cell-type dependent in ACCs in the lacrimal gland, while HPV infection was negative. P16 overexpression was unrelated to HPV infection. The mechanism of p16 overexpression needs to be further investigated in ACCs in the lacrimal gland.

Clinicopathologic Assessment of Ocular Adnexal Lymphoproliferative Lesions at a Tertiary Eye Hospital in Iran.

BACKGROUND: The most common type of ocular lymphoma is non-Hodgkin lymphoma (NHL), categorized into two groups: indolent (slow growing) and aggressive (rapid growing). Differentiating benign reactive lymphoid hyperplasia (RLH) from malignant ocular adnexal lymphoma (OAL) is challenging. Histopathology, immunohistochemistry (IHC) and ow cytometry have been used as diagnostic tools in such cases. MATERIALS AND METHODS: In this retrospective case series, from 2002 to 2013 at Farabi Eye Center, 110 patients with ocular lymphoproliferative disease were enrolled. Prevalence, anatomical locations, mean age at diagnosis and the nal diagnosis of the disease with IHC were assessed. Comparison between previous pathologic diagnoses and results of IHC was made. Immunoglobulin light chains and B-cell and T-cell markers and other immuno-phenotyping markers including CD20, CD3, CD5, CD23, CD10, CYCLIND1 and BCL2 were evaluated to determine the most accurate diagnosis. The lymphomas were categorized based on revised European-American lymphoma (REAL) classi cation. RESULTS: Mean age±SD (years) of the patients was 55.6 ±19.3 and 61% were male. Patients with follicular lymphoma, large B-cell lymphoma or chronic lymphocytic leukemia/small cell lymphoma (CLL/SLL) tended to be older. Nine patients with previous diagnoses of low grade B-cell lymphoma were re-evaluated by IHC and the new diagnoses were as follows: extranodal marginal zone lymphoma(EMZL) (n=1), SLL(n=1), mantle cell lymphoma (MCL) (n=3), reactive lymphoid hyperplasia RLH (n=2). Two cases were excluded due to poor blocks. Flow cytometry reports in these seven patients revealed SLL with positive CD5 and CD23, MCL with positive CD5 and CyclinD1 and negative CD23, EMZL with negative CD5,CD23 and CD10. One RLH patient was negative for Kappa/Lambda and positive for CD3 and CD20 and the other was positive for all of the light chains, CD3 and CD20. Orbit (49.1%), conjunctiva (16.1%) and lacrimal glands (16.1%) were the most common sites of involvement. CONCLUSIONS: Accurate pathological classi cation of lesions is crucial to determine proper therapeutic approaches. This can be achieved through precise histologic and IHC analyses by expert pathologists.

The cost of ipilimumab toxicity: a single-centre analysis.

There are no published data on the costs associated with investigating and managing toxicity from ipilimumab treatment in patients with metastatic melanoma. Patients treated with ipilimumab at The Royal Marsden Hospital between 1 September 2010 and 1 April 2013 were identified. Data on demographics, investigations and survival outcomes were collected. Patients with grade 3 or higher immune-related adverse events were identified, and costs of investigating and managing toxicities in them were calculated on the basis of standard National Health Service tariffs. Out of the 110 patients, 29 experienced grade 3/4 immune-related adverse events. The total cost of investigating and managing these patients was £140,680, or a median cost of £2860 per patient. Patients experiencing grade 3/4 toxicities had 1-, 2- and 3-year survival rates of 79, 59 and 46%, compared with 24, 17 and 15% in the group that did not experience significant toxicity (P<0.0005). The most common treatment-related toxicity identified was colitis. Two patients died from complications associated with ipilimumab colitis. The cost of ipilimumab toxicity is marginal in comparison with the total treatment cost. Patients treated with ipilimumab who develop significant toxicity have a higher than expected 1-, 2- and 3-year survival.

[Ophthalmological imaging with ultrahigh field magnetic resonance tomography: technical innovations and frontier applications]. / Ophthalmologische Bildgebung mit Ultrahochfeld-Magnetresonanztomografie: technische Innovationen und wegweisende Anwendungen.

This review documents technical progress in ophthalmic magnetic resonance imaging (MRI) at ultrahigh fields (UHF, B(0) ≥ 7.0 T). The review surveys frontier applications of UHF-MRI tailored for high spatial resolution in vivo imaging of the eye, orbit and optic nerve. Early examples of clinical ophthalmic UHF-MRI including the assessment of melanoma of the choroid membrane and the characterisation of intraocular masses are demonstrated. A concluding section ventures a glance beyond the horizon and explores research promises along with future directions of ophthalmic UHF-MRI.

Method for verifying the air kerma strength of I-125 plaques for the treatment of ocular melanoma.

The purpose of this work was to develop a method for easily verifying that the activity or air kerma strength of pre-assembled eye plaques, used in the treatment of ocular melanomas, agrees with the activity or air kerma strength called for in the treatment plan. A Capintec CRC-7 Dose Calibrator with its standard vial/syringe sample holder was used to measure the activity of pre-assembled COMS and Eye Physics EP917 eye plaques using IsoAid Advantage I-125 seeds. Plaque activity measurements were made by placing the plaque face up in the center of a 5 cm tall Styrofoam insert in the source holder. Activity measurements were made with the source holder rotated to four angles (0°, 90°, 180°, and 270°). The average of these four values was converted to air kerma strength and divided by the assay air kerma strength, from the NIST traceable source calibration, and decayed to the plaque measurement date, to determine a plaque calibration factor. The average of the calibration factors for each plaque type was used to establish a calibration factor for each plaque type. Several partially loaded plaque configurations were included in this study and different methods were used to determine the effects of partial loading. This verification method is easy to implement with commonly available equipment and is effective in identifying possible errors. During this two-year study, the air kerma strength of 115 eye plaques was checked and 11 possible errors were identified.

Ocular adnexal lymphoma: assessment of a tumor-node-metastasis staging system.

PURPOSE: To assess distribution, correlations, and prognostic effect of tumor (T), node (N), and metastasis (M) staging on relapse and survival. DESIGN: Retrospective clinical review. PARTICIPANTS: Sixty-three patients diagnosed with primary ocular adnexal lymphoma (OAL) between January 1986 and November 2011. METHODS: Complete ocular examination and systemic evaluation were performed. Patients were staged according to the American Joint Committee on Cancer (AJCC) seventh edition tumor-node-metastasis (TNM) clinical staging system for OAL and followed every 6 to 12 months (median follow-up, 27.9 months). MAIN OUTCOME MEASURES: Relapse defined as lymphoma recurrence in the initial site of presentation, the contralateral ocular adnexal structures, or other systemic site and overall survival. RESULTS: There were 40 men (63.5%). The median age was 65 years (range, 24-85 years). The affected site was the conjunctiva in 27 patients (42.9%), orbit in 38 patients (60.3%), and eyelid in 3 patients (4.8%). The histologic subtype was extranodal marginal zone lymphoma (EMZL) in 51 patients (81.0%). A total of 14 patients (23.3%) had T1, 42 patients (70.0%) had T2, 1 patient (1.7%) had T3, and 3 patients (5.0%) had T4 disease. A total of 48 patients (82.8%) had N0 disease, and 10 patients (17.2%) had N1-4 disease. M stage was M0 in 47 patients (81.0%) and M1 in 11 patients (19.0%). With advanced T stage, there was an increase in both N1-4 (P = 0.045) and M1 disease (P = 0.041). M1 disease was greater among patients with N1-4 disease compared with N0 stage (50.0% vs. 12.5%, P = 0.003). Overall, 18 patients (28.6%) relapsed and 6 patients (9.5%) died. In Cox analysis, relapse was not associated with T stage (hazard ratio [HR], 1.14 per 1 level increase, P = 0.71), N stage (HR, 1.47; P = 0.51 N1-4 vs. N0), or M stage (HR, 1.22; P = 0.76 M1 vs. M0). T stage was not associated with survival (HR, 0.86; P = 0.81), whereas N1-4 had marginally worse survival than N0 (HR, 5.35; P = 0.07), and M1 had worse survival than M0 (HR, 9.27; P = 0.008). CONCLUSIONS: The TNM staging system for primary OAL is useful for precise characterization of extent of local disease. Although T stage does not predict relapse or survival, N1-4 and M1 stages indicated less favorable survival. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Assessment of dynamic contrast-enhanced magnetic resonance imaging in the differentiation of malignant from benign orbital masses.

OBJECTIVE: Dynamic contrast enhanced MR imaging (DCE-MRI) allows imaging of the physiology of the microcirculation. The purpose of this study was to determine the diagnostic efficacy of time intensity curve (TIC) and DCE parameters for characterization of orbital masses. METHODS: Fifty-nine patients with untreated orbital lesions underwent DCE-MRI before surgery. For each lesion, peak height (PH), maximum enhancement ratio (ERmax), time of peak enhancement (Tpeak) and maximum rise slope (Slopemax) were plotted and calculated. Receiver operator characteristics (ROC) analysis was conducted to assess the appropriate cut-off value. RESULTS: All 26 lesions that demonstrated persistent pattern (type-I) TICs were benign. Most of the masses with the washout pattern (type-III) TIC were malignant (10/14), including lymphoma (n=6) and melanoma (n=4). The Slopemax of benign lesions was statistically lower than malignant ones, while the ERmax and Tpeak values of benign lesions were significantly higher. No statistical difference was found in PH (P=0.121). The AUC for ERmax, Tpeak and Slopemax in differentiating benign orbital lesions from malignant ones were 0.683, 0.837 and 0.738, respectively. In the three DCE parameters, Slopemax cut-off value of 1.10 provided the highest sensitivity of 93.8%; however, the corresponding specificity was low (58.1%). The ERmax cut-off value of 1.37 and Tpeak cut-off value of 35.14 respectively offered the best diagnostic performances. CONCLUSION: DCE-MRI, especially the qualitative TIC pattern and quantitative value of Slopemax, ERmax and Tpeak, could be a complementary investigation in distinguishing malignant orbital tumor from benign ones.

Trabecular meshwork length in men and women by histological assessment.

PURPOSE: To evaluate the length of the trabecular meshwork (TM) from the scleral spur (SS) to Schwalbe's line (SL) and assess the detectability of the SS in histopathology specimens. METHODS: This study included 158 angle images from 79 cross-sectional slides derived from eyes enucleated for melanoma. The slides were stained with hematoxylin-eosin (HE) or periodic acid schiff (PAS). Two ophthalmologists evaluated the TM length by using the slides stained with HE to assess the interobserver reproducibility. For intraobserver reproducibility, the first observer assessed 79 images in a different session. Also, 30 images that were randomly selected for PAS stain were evaluated to assess the agreement of the measurements between HE and PAS staining. Interclass correlation coefficients (ICC) were calculated to evaluate reproducibility of measurements. The images were also evaluated for detectability of the scleral spurs. RESULTS: Among the 79 included subjects, 40 were male and 39 were female. The average trabecular meshwork length was 694.9 ± 109.0 µm in the male group and 713.2 ± 106.9 µm in the female group (p = 0.29). Intraobserver and interobserver ICC were 0.89 and 0.62, respectively. ICC for agreement between HE and PAS was 0.89. Among the 158 angles graded, the first observer graded 40 images (25.3%) and the second observer graded 45 (28.5%) as difficult to identify the scleral spur. CONCLUSIONS: There was no statistically significant difference between the average trabecular meshwork length in men and women. Among the angles evaluated, 25.3-28.5% were graded as difficult to identify the scleral spur.

Dosimetry of (125)I and (103)Pd COMS eye plaques for intraocular tumors: report of Task Group 129 by the AAPM and ABS.

Dosimetry of eye plaques for ocular tumors presents unique challenges in brachytherapy. The challenges in accurate dosimetry are in part related to the steep dose gradient in the tumor and critical structures that are within millimeters of radioactive sources. In most clinical applications, calculations of dose distributions around eye plaques assume a homogenous water medium and full scatter conditions. Recent Monte Carlo (MC)-based eye-plaque dosimetry simulations have demonstrated that the perturbation effects of heterogeneous materials in eye plaques, including the gold-alloy backing and Silastic insert, can be calculated with reasonable accuracy. Even additional levels of complexity introduced through the use of gold foil "seed-guides" and custom-designed plaques can be calculated accurately using modern MC techniques. Simulations accounting for the aforementioned complexities indicate dose discrepancies exceeding a factor of ten to selected critical structures compared to conventional dose calculations. Task Group 129 was formed to review the literature; re-examine the current dosimetry calculation formalism; and make recommendations for eye-plaque dosimetry, including evaluation of brachytherapy source dosimetry parameters and heterogeneity correction factors. A literature review identified modern assessments of dose calculations for Collaborative Ocular Melanoma Study (COMS) design plaques, including MC analyses and an intercomparison of treatment planning systems (TPS) detailing differences between homogeneous and heterogeneous plaque calculations using the American Association of Physicists in Medicine (AAPM) TG-43U1 brachytherapy dosimetry formalism and MC techniques. This review identified that a commonly used prescription dose of 85 Gy at 5 mm depth in homogeneous medium delivers about 75 Gy and 69 Gy at the same 5 mm depth for specific (125)I and (103)Pd sources, respectively, when accounting for COMS plaque heterogeneities. Thus, the adoption of heterogeneous dose calculation methods in clinical practice would result in dose differences >10% and warrant a careful evaluation of the corresponding changes in prescription doses. Doses to normal ocular structures vary with choice of radionuclide, plaque location, and prescription depth, such that further dosimetric evaluations of the adoption of MC-based dosimetry methods are needed. The AAPM and American Brachytherapy Society (ABS) recommend that clinical medical physicists should make concurrent estimates of heterogeneity-corrected delivered dose using the information in this report's tables to prepare for brachytherapy TPS that can account for material heterogeneities and for a transition to heterogeneity-corrected prescriptive goals. It is recommended that brachytherapy TPS vendors include material heterogeneity corrections in their systems and take steps to integrate planned plaque localization and image guidance. In the interim, before the availability of commercial MC-based brachytherapy TPS, it is recommended that clinical medical physicists use the line-source approximation in homogeneous water medium and the 2D AAPM TG-43U1 dosimetry formalism and brachytherapy source dosimetry parameter datasets for treatment planning calculations. Furthermore, this report includes quality management program recommendations for eye-plaque brachytherapy.

Radiobiology for eye plaque brachytherapy and evaluation of implant duration and radionuclide choice using an objective function.

PURPOSE: Clinical optimization of Collaborative Ocular Melanoma Study (COMS) eye plaque brachytherapy is currently limited to tumor coverage, consensus prescription dosage, and dose calculations to ocular structures. The biologically effective dose (BED) of temporary brachytherapy treatments is a function of both chosen radionuclide R and implant duration T. This study endeavored to evaluate BED delivered to the tumor volume and surrounding ocular structures as a function of plaque position P, prescription dose, R, and T. METHODS: Plaque-heterogeneity-corrected dose distributions were generated with MCNP5 for the range of currently available COMS plaques loaded with sources using three available low-energy radionuclides. These physical dose distributions were imported into the PINNACLE(3) treatment planning system using the TG-43 hybrid technique and used to generate dose volume histograms for a T = 7 day implant within a reference eye geometry including the ciliary body, cornea, eyelid, foveola, lacrimal gland, lens, optic disc, optic nerve, retina, and tumor at eight standard treatment positions. The equation of Dale and Jones was employed to create biologically effective dose volume histograms (BEDVHs), allowing for BED volumetric analysis of all ROIs. Isobiologically effective prescription doses were calculated for T = 5 days down to 0.01 days, with BEDVHs subsequently generated for all ROIs using correspondingly reduced prescription doses. Objective functions were created to evaluate the BEDVHs as a function of R and T. These objective functions are mathematically accessible and sufficiently general to be applied to temporary or permanent brachytherapy implants for a variety of disease sites. RESULTS: Reducing T from 7 to 0.01 days for a 10 mm plaque produced an average BED benefit of 26%, 20%, and 17% for (103)Pd, (125)I, and (131)Cs, respectively, for all P; 16 and 22 mm plaque results were more position-dependent. (103)Pd produced a 16%-35% BED benefit over (125)I, whereas (131)Cs produced a 3%-7% BED detriment, independent of P, T, and plaque size. Additionally, corresponding organ at risk physical doses were lowest using (103)Pd in all circumstances. CONCLUSIONS: The results suggest that shorter implant durations may correlate with more favorable outcomes compared to 7 day implants when treating small or medium intraocular lesions. The data also indicate that implant duration may be safely reduced if the prescription physical dose is likewise diminished and that (103)Pd offers a substantial radiobiological benefit over (125)I and (131)Cs irrespective of plaque position, implant duration, and tumor size.

Bone invasion by adenoid cystic carcinoma of the lacrimal gland: preoperative imaging assessment and surgical considerations.

PURPOSE: To identify the incidence of radiologically and histologically documented bony invasion of the lacrimal gland fossa by adenoid cystic carcinoma. PATIENTS AND METHODS: The authors reviewed the records of all 18 patients with lacrimal gland adenoid cystic carcinoma surgically treated at their institution from 1997 to 2009 for imaging findings (blinded review) and histologic findings on evaluation of the lacrimal gland fossa. Preoperative CT and/or MRI findings were available for 17 patients. RESULTS: The 8 men and 10 women ranged in age from 9 to 69 years. American Joint Committee on Cancer tumor stages after preoperative imaging were as follows: T1N0M0, 2 patients; T2N0M0, 5 patients; T3aN0M0, 2 patients; T3bN0M0, 5 patients; T3bN0M1, 2 patients; T4bN0M0, one patient; and TxN0M0, one patient. Preoperative imaging suggested bony involvement of the lacrimal gland fossa in 13 patients (76.5%); this was histologically confirmed in 11 of the 13. Preoperative imaging suggested no bone involvement in 4 patients, 3 of whom had bone involvement by histology. Overall, 14 of 17 histologically evaluable cases (82.3%) had invasion of the lacrimal gland fossa. Histologic findings of bone/periosteal involvement led to upstaging of 3 tumors. Metastases developed in 8 of 18 patients and trended with basaloid histology (p = 0.066). CONCLUSIONS: Adenoid cystic carcinoma of the lacrimal gland is associated with bone invasion in essentially all but the smallest of tumors (T1). This high rate of bone involvement may warrant addressing the bony walls during surgery for adenoid cystic carcinoma of the lacrimal gland.

A parameter study of pencil beam proton dose distributions for the treatment of ocular melanoma utilizing spot scanning.

The results of Monte Carlo calculated dose distributions of proton treatment of ocular melanoma are presented. An efficient spot scanning method utilizing active energy modulation, which also minimizes the number of target spots was developed. We simulated various parameter values for the particle energy spread and the pencil beam diameter in order to determine values suitable for medical treatment. We found that a 2.5-mm-diameter proton beam with a 5% Gaussian energy spread was suitable for treatment of ocular melanoma while preserving vision for the typical case that we simulated. The energy spectra and the required proton current were also calculated and are reported. The results are intended to serve as a guideline for a new class of low-cost, compact accelerators.

Assessment of HOPE fixation in vitrectomy specimens in patients with chronic bilateral uveitis (masquerade syndrome).

BACKGROUND: Cytological examination of rapidly transported, unfixed vitreous specimens is considered the gold standard in exclusion of primary intraocular lymphoma (PIOL) in patients with idiopathic steroid resistant chronic uveitis. These specimens are difficult to interpret, and reports of "false negatives" or "false positives" are common. Fixation using HOPE solution (Herpes-glutamic acid buffer mediated Organic solvent Protection Effect) has been successfully applied in the investigation of cytospin preparations using immunocytology, in situ hybridisation and polymerase chain reaction (PCR). The purpose of this study was to compare the cytologic features of vitrectomy specimens from patients with clinical reactive vitritis and PIOL prepared following HOPE fixation with those in conventional cytospins and to identify any diagnostic pitfalls. METHODS: Pars plana vitrectomy was performed in 15 cases of patients with chronic uveitis. The vitreous samples were halved: one half was fixed in HOPE solution; the other half remained unfixed. All samples were subject to conventional staining, immunocytology and clonality analysis using polymerase chain reaction (IgH-PCR) and GeneScan. The specimens were assessed following by two pathologists who were masked to the cytological preparation method. The specimens were evaluated for cellularity, cellular appearance, cytoplasmic and nuclear features as well as quality of the immunostains. RESULTS: Twelve of the fifteen vitreous samples were diagnosed as reactive vitritis; in three specimens, a primary intraocular lymphoma of B-cell type was diagnosed. Compared to the unfixed vitreal specimens, the quality of the cytomorphology and immunocytology improved in the HOPE-fixed specimens. IgH-PCR and GeneScan analysis demonstrated polyclonal amplification products in the reactive cases, and monoclonal B-cell populations in the B-PIOL. CONCLUSION: Our results demonstrate that cytomorphology and immunoreactivity of vitreous specimens are well preserved following HOPE fixation. DNA of sufficient quality could be extracted from HOPE-fixed vitreous biopsies, in order to perform clonality analyses. HOPE fixation appears to be promising in simplifying the transportation of these specimens, and may improve the diagnostic reliability of vitreous specimens in patients with masquerade syndrome.

Quantitative detection of circulating tumor cells in cutaneous and ocular melanoma and quality assessment by real-time reverse transcriptase-polymerase chain reaction.

PURPOSE: Inconsistent reports on the detection of melanoma cells in peripheral blood by reverse transcriptase-PCR (RT-PCR) have resulted in uncertainty on the prognostic value of circulating melanoma cells. EXPERIMENTAL DESIGN: We developed real-time RT-PCR assays for quantitation of tyrosinase, MelanA/MART1, and gp100 and for porphobilinogen deaminase housekeeping gene. Melanoma tissue (n = 18), peripheral blood samples from healthy donors (n = 21), and patients with cutaneous (n = 122) and uveal (n = 64) melanoma from our institution were analyzed. For quality control, an additional 251 samples from ongoing multicenter studies were compared with in-house samples. RESULTS: Tyrosinase was not detected in healthy donor blood samples. For the two other markers, cutoff values had to be defined to distinct patient samples from controls. Patients with stage IV uveal and cutaneous melanoma expressed all three markers more frequently and at higher levels in peripheral blood as compared with earlier stages. The variation of expression was 4 logs and correlated with tumor load and serum lactate dehydrogenase. In 2 of 3 uveal melanoma patients, detection of circulating tumor cells preceded the development of liver metastases. The diagnostic sensitivity was optimal in blood samples containing >0.1pg/ microl porphobilinogen deaminase (95.7% of in-house samples and 57.4% of multicenter samples). CONCLUSIONS: Real-time RT-PCR is able to quantitatively define the quality of a sample and provides quantitative data for melanoma markers. Disparities in the results of previous studies may be attributable to undetected differences in sample quality. The prognostic relevance of this assay is currently under evaluation in several prospective randomized trials.

Incidence and management of secondary malignancies in patients with retinoblastoma and Ewing's sarcoma.

Childhood cancer survivors at highest risk of developing a secondary malignancy are those with hereditary retinoblastoma. The majority of such secondary cancers will be sarcomas, most commonly of bone. One-third of these occur outside a typical radiation field, commonly in an extremity. Bone sarcoma is also the most commonly reported secondary cancer to develop among survivors of Ewing's sarcoma. In this group, radiation doses greater than 60 Gy as well as alkylating agent chemotherapy have been identified as contributors to the increased risk. The prognosis for patients with a secondary sarcoma has been poor, with few cures reported to date. However, an aggressive, combined modality approach, including radical resection, postoperative radiation, and adjuvant chemotherapy, may improve the survival rate.

Bromodeoxyuridine uptake in the assessment of hyperthermic therapy for intraocular tumor.

Assessment of uveal melanoma response after many nonenucleation therapies is difficult with current modalities, since tumor regression is usually delayed. The goal of most treatments, including ionizing radiation or radiation and adjunct hyperthermia, is to destroy the reproductive capacity of the tumor. Cell cycling analysis with bromodeoxyuridine, a thymidine analogue only incorporated during DNA synthesis, was a useful indication of tumor control after hyperthermia was used to treat a Greene intraocular melanoma model. Cell cycling decreased from a mean of 16% before therapy to less than 1% in all the successfully treated tumors. Cell cycling changes preceded histologic evidence of cell death. In contrast, tumors that grew after insufficient treatment had increased cell cycling to a mean of 25%. Cell cycling studies with bromodeoxyuridine represent a sensitive gauge of the reproductive integrity of the tumor.

CT-scan and intraocular tumours: detection and assessment of size and extrascleral growth of uveal melanomas.

Twenty-two patients, who were suspected of an intraocular tumour, were examined with the high resolution thin slice CT-scan (1.5 or 3 mm slices). The final diagnosis in 14 cases was an uveal melanoma, and in 3 cases in which an uveal melanoma was suspected a chorioretinal haemorrhage, an exudative macular degeneration type Junius Kuhnt and an intrascleral foreign body were respectively found. In 2 cases intraocular metastases were demonstrated, and in 2 other cases malignant lymphomata. One patient had a haemangioma. In 10 of the 14 patients with uveal melanomata enucleation was performed and 4 were treated by ruthenium application. The CT-scan was compared with ultrasonographic, operative and histological findings. From the study it appears that the CT-scan is useful for screening uveal melanomas. It is however difficult to distinguish the uveal melanoma from a secondary retinal detachment; differentiation from an exudative macular degeneration was not possible. The size of the tumour was only represented correctly on the CT-scan in half the cases. CT examination fails to demonstrate or exclude epi- and extrascleral growth. The multiple metastases and the haemangioma could only be seen with difficulty on the CT-scan; the latter was however clearly visible after the injection of contrast. The foreign body could be clearly seen without contrast.