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Epithelial thinning and breaks in Bowman’s layer are the most common histopathological findings in keratoconic explanted corneas. However, histopathological changes can also be observed in all five layers of the cornea. Compaction of deep stromal layers and fold or breaks in Descemet’s membrane are associated with severe keratoconus. This chapter provides an overview of histopathological findings in explanted corneas of keratoconus as well as guidelines for evaluating a corneal specimen macro- and microscopically.

There is currently no clinical risk-assessment tool allowing identification of patients at risk for developing uveal melanoma (UM) who might benefit from regular screening. As a first step toward the elaboration of such a tool, we systematically reviewed UM risk factors already established by meta-analysis.Two reviewers independently screened Pubmed, Medline, Embase, and Web of Science from their respective inception dates until July 2016 using a combination of keywords and MeSH terms. Eligible studies were meta-analyses or systematic reviews providing pooled odds ratios (ORs) of risk factors for UM development or sufficient information to calculate them. Methodological quality was evaluated using the Assessment of Multiple Systematic Reviews tool.Four meta-analyses with a mean methodological quality score of 65.9% (min: 54.5%; max: 72.7%) were included. The following significant risk factors were identified: atypical cutaneous nevi (OR 2.82, 95% CI 1.10-7.26), welding (OR 2.05, 95% CI 1.20-3.51), occupational cooking (OR 1.81, 95% CI 1.33-2.46), fair skin color (OR 1.80, 95% CI 1.31-2.47), light eye color (OR 1.75, 95% CI 1.31-2.34), common cutaneous nevi (OR 1.74, 95% CI 1.27-2.39), propensity to sunburn (OR 1.64, 95% CI 1.29-2.09), iris nevi (OR 1.53, 95% CI 1.03-2.27), and cutaneous freckles (OR 1.27, 95% CI 1.09-1.49). Non-significant factors included outdoor leisure activity, occupational sunlight exposure, latitude of birth, and hair color.Moderate quality of evidence determined nine significant risk factors for developing UM. Knowledge of these variables will assist researchers in the elaboration of a formal risk-assessment tool allowing clinicians to estimate susceptibility to the disease and necessity of regular screening.

Purpose The aim of this study was to determine the frequency of clinically unsuspected ocular surface squamous neoplasia (OSSN) in cases of biopsied pterygium (PT). Methods We reviewed 15,016 cases presented at the Henry C. Witelson Ocular Pathology Laboratory during the period 1993–2013. All cases with a clinical diagnosis of PT were included. Histopathological diagnoses were reviewed and demographic data were retrieved from histopathological request forms. All cases associated with OSSN were re-evaluated independently by two ocular pathologists. The classification of OSSN in PT cases was made based on the Armed Forces Institute of Pathology (AFIP) recommendations. Results Two hundred and fifteen cases were diagnosed clinically as PT (1.43%) and 54% were from male patients. The average age at diagnosis was 53.4±15.5years. OSSN was identified in five cases (2.33%), and four of these cases were from female patients (80%). The average age of patients with PT and OSSN was similar to PT patients without OSSN ( P >0.05). Cases with OSSN were diagnosed as conjunctival intraepithelial neoplasia (CIN) I (60%), CIN II (20%), and CIN III (20%). There was complete agreement between the two pathologists (100%). Conclusions The relatively high rate of dysplasia in a low ultraviolet light index area challenges the main cause of this disease in our population, a hypothesis that should be evaluated in future studies. We suggest that all PT samples should be sent for histopathological evaluation even in areas with low ultraviolet light index.

Objectives: The aim of this study was to evaluate heat shock protein 90 (HSP90) expression in squamous lesions (SLs) and to assess its diagnostic value for different lesions within the SL spectrum. Methods: A total of 70 conjunctival SLs, including 19 papillomas, 22 cases of conjunctival intraepithelial neoplasia (ConINs) I, 11 cases of ConIN II, six cases of ConIN III, and 12 squamous carcinomas (sqCAs), were evaluated using the German immunoreactive score against HSP90. Results: Cytoplasmic HSP90 expression differed between low- and high-grade lesions ( P < .001). Among high-grade lesions, the nuclear HSP90 score was higher in the ConIN III–sqCA group than in the ConIN II group ( P = .0162). A percentage of total thickness staining of less than 73% differentiated between ConIN III and sqCA. Conclusions: The expression of HSP90 is particularly useful to differentiate low-grade from high-grade lesions of the conjunctiva. HSP90 may play an important role in the malignant transformation of SLs and could be a new target for therapy.

Background. Ocular involvement in mycosis fungoides (MF) cases occurs in one-third of patients with the eyelid being the most frequent site affected; however, conjunctival involvement is rarely reported. Herein, we report a rare case of conjunctival involvement of MF.Case Presentation. A 66-year-old man who was previously diagnosed with MF in 2010 and was treated presented in 2014 complaining of foreign body sensation and redness in both eyes. Slit lamp examination of both eyes showed erythematous conjunctival growth that extended circumferentially. Physical examination revealed erythematous skin lesions on different body parts. Conjunctival biopsy was performed and revealed a dense, highly polymorphic lymphocytic population. The immunophenotype demonstrated a neoplastic T-cell origin consistent with MF. A diagnosis of conjunctival involvement by MF was made. The conjunctiva was treated with radiotherapy resulting in tumor regression. There were no recurrences at the 6-month follow-up.Conclusion. T-cell lymphoma should be considered in patients with a history of MF presenting with conjunctival and skin lesions.

The �耀eld of wilderness therapy has placed increasing importance on measuring effectiveness. Though studies demonstrate positive outcomes for adolescents, we lack representative samples, post-discharge data, and replication of positive results. This three-year study sought to measure outcomes and to identify mechanisms of change in wilderness therapy. We found statistically signi�耀cant change from intake to discharge on the Youth Outcome Questionnaire ® and on measures of hope, life effectiveness, and treatment expectancy. We discuss when change occurs as well as mechanisms of change, demographic differences, and critical lessons we learned about conducting research in a clinical setting.

Purpose/aim: Posterior capsular opacification (PCO) is the leading complication following cataract surgery. However, PCO grading methods vary between studies and rely on subjective scoring methods; hence, they are prone to error. Therefore, software that reduces subjectivity and enhances reproducibility is needed. The aim of the study was to evaluate the reproducibility and objectivity of a novel automated custom-designed PCO detection software.In total, 165 fixed eyes with intraocular lenses were sectioned to obtain a Miyake Apple view (MAV) image. The capsular bag (CB) with the in situ IOL was removed and imaged using the Olympus DSX110 stereoscope. Central, peripheral, and Soemmering's ring area (SRA) and intensity (SRI) were graded by two ophthalmologists using a scale of 0-4. Software was developed to detect PCO, and these values were correlated with subjective scoring.Intra-observer agreement for MAV and CB images ranged from poor to moderate, with very good agreement for SRI (k = 0.88, 0.86, respectively). Agreement between graders was fair to good (k = 0.31-0.77). Miyake Apple view and capsular bag images showed good correlation; central PCO showed the least correlation for both evaluators (k = 0.29, 0.72). Subjective average grading versus software correlation demonstrated very good correlation for intensity and good for area (r = 0.85, 0.61). Reproducibility of the methodology resulted in good to very good correlation.Software correlates with previous scoring methods and is a reliable and reproducible system. Moreover, the capsular bag view, as opposed to the Miyake Apple view, allows visualization of the capsular bag directly and avoids overlapping structures (vitreous, cornea) that may interfere with proper PCO quantification.

Purpose: To evaluate the effects of ranibizumab and amfenac in human uveal melanoma cell lines and to explore the ability of these compounds to sensitize uveal melanoma cells to radiation therapy. Methods: The 92.1 human uveal melanoma cell line was cultured and subjected to the proposed treatment (ranibizumab, amfenac, and a combination of both). Proliferation, migration, and invasion assays of the 92.1 uveal melanoma cell line were assessed after pretreatment with ranibizumab (125 mg/mL), amfenac (150 nM), or a combination of both. In addition, proliferation rates were assessed after treatment with ranibizumab and amfenac, and the cells were subsequently exposed to various radiation doses (0, 4, and 8 Gy). Results: Proliferation assay: cells treated with a combination of ranibizumab and amfenac had lower proliferation rates than controls (p=0.016) and than those treated with only ranibizumab (p=0.033). Migration assay: a significantly lower migration rate was observed in cells treated with amfenac than the control (p=0.014) and than those treated with ranibizumab (p=0.044). Invasion assay: there were no significant differences among the studied groups. Irradiation exposure: in the 4 Gy dose group, there were no significant differences among any groups. In the 8 Gy dose group, treatment with ranibizumab, amfenac, and their combination prior to application of the 8 Gy radiation led to a marked reduction in proliferation rates (p=0.009, p=0.01, and p=0.034, respectively) compared with controls. Conclusion: Combination of ranibizumab and amfenac reduced the proliferation rate of uveal melanoma cells; however, only amfenac monotherapy significantly decreased cell migration. The radiosensitivity of the 92.1 uveal melanoma cell line increased following the administration of ranibizumab, amfenac, and their combination. Further investigation is warranted to determine if this is a viable pretreatment strategy to render large tumors amenable to radiotherapy. RESUMO Objetivo: Avaliar os efeitos do ranibizumabe em associação com o amfenac nas células de melanoma uveal humano e explorar a capacidade desses compostos em sensibilizar as células de melanoma uveal à radioterapia. Métodos: Células de melanoma uveal humano do tipo 92.1 foram cultivadas e submetidas ao tratamento proposto (ranibizumabe, amfenac e a combinação de ambos). Ensaios de proliferação, migração e invasão com as células de melanoma uveal do tipo 92.1 foram avaliados após tratamento com ranibizumabe (125 mg/ml), amfenac (150 nM) e a combinação de ambos. Além disso, as taxas de proliferação foram avaliadas após tratamento com ranibizumabe e amfenac com subsequente exposição das células a diferentes doses de radiação (0 Gy, 4 Gy e 8 Gy). Resultados: Ensaio de proliferação: células tratadas com ranibizumabe e amfenac combinados apresentaram taxas de proliferação inferiores em comparação ao grupo controle (p=0,016), do que as tratadas apenas com ranibizumabe (p=0,033). Ensaio de migração: foi observada uma taxa de migração significativamente mais baixa nas células tratadas com amfenac do que no grupo controle (p=0,014) e do que nas tratadas com ranibizumabe (p=0,044). Ensaio de invasão: não houve diferenças significativas entre os grupos estudados. Exposição à irradiação: no grupo da dose de 4 Gy, não houve diferença significante entre os grupos. No grupo da dose de 8 Gy, o tratamento com ranibizumabe, afenac e sua combinação antes da aplicação da radiação de 8 Gy levou a uma redução acentuada nas taxas de proliferação (p=0,009, p=0,01 e p=0,034, respectivamente) em comparação aos grupos controle. Conclusão: A combinação de ranibizumabe e amfenac reduziu a taxa de proliferação das células de melanoma uveal; no entanto, apenas o amfenac diminuiu significativamente a migração celular. A radiossensibilidade das células de melanoma uveal do tipo 92.1 aumentou após a administração de ranibizumabe, amfenac e sua combinação. Mais investigações são necessárias para determinar se esta é uma estratégia de pré-tratamento viável para tornar grandes tumores passíveis de radioterapia.

Purpose: To investigate the effect of nicotinamide on the secretion of pro-an giogenic and pro-inflammatory cytokines in uveal melanoma cell lines. Methods: Two human uveal melanoma cell lines (92.1 and OCM-1) were treated with nicotinamide (10 mmol/L) or control media for 48 hours in culture. The su perna tant from each culture was used in sandwich enzyme-linked immuno sorbent assay-based angiogenesis and inflammation arrays to evaluate the effects of exogenously administered nicotinamide on the secretion of a total of 20 pro-an gio genic and pro-inflammatory proteins. Results: Seven pro-angiogenic cytokines were detected under control conditions for both uveal melanoma cell lines. Treatment with nicotinamide resulted in a significant decrease in secretion of the following pro-angiogenic cytokines: angiogenin, angiopoietin-2, epidermal growth factor, and vascular epithelial growth factor-A in the 92.1 cells; basic fibroblast growth factor in the OCM-1 cells; and placenta growth factor in both cell lines. Among the pro-inflammatory proteins, monocyte chemotactic protein-1 and interleukin-8 were expressed in both untreated cell lines and both were significantly reduced when treated with nicotinamide. Conclusions: Results from this in vitro model suggest that nicotinamide may have anti-inflammatory and anti-angiogenic properties, which may open the possibility of using it as a chemopreventive agent for uveal melanoma; however, further studies including animal models are warranted. RESUMO Objetivo: Acredita-se que a nicotinamida (NIC) seja capaz de diminuir a angiogênese induzida pelo fator de crescimento endotelial vascular (VEGF). Investigar os efeitos da nicotinamida sobre a secreção de citocinas pró-angiogênicas e pró-inflamatórias em linhagens de células de melanoma uveal humano (UM). Métodos: Duas linhagens de células humanas de UM (92,1 e OCM-1) foram tratadas com NIC (10 mmol/L) ou apenas com meio de cultura por 48 horas. O sobrenadante das culturas obtido após a administração de nicotinamida foi comparado com o sobrenadante das culturas controle quanto à expressão de 20 fatores pró-angiogênicos e pró-inflamatórios, pela técnica de enzyme-linked immunosorbent assay (ELISA). Resultados: Sete citocinas pró-angiogênicas foram detectadas nas condições de controle em ambas as linhagens de células de UM. O tratamento com nicotinamida promoveu uma redução significativa da secreção das seguintes citocinas angiogênicas: Angiogenina, ANG2, EGF e VEGF-A em células 92.1; bFGF em células OCM-1; PIGF em ambas as linhagens celulares. Quanto às proteínas pró-inflamatórias, a expressão de MCP-1 e IL-8 foi significativamente reduzida com a administração de nicotinamida em relação às culturas de células que não receberam o tratamento. Conclusões: Nicotinamida apresenta propriedades anti-inflamatórias e anti-angiogênicas em modelo experimental in vitro. Tais efeitos sugerem a possibilidade de utilizar esta substância na quimioprevenção do UM. Entretanto, estudos com modelos experimentais in vivo são necessários para melhor avaliar o benefício do tratamento do UM com nicotinamida.

There has been considerable growth in outcomes research with adolescent clients in wilderness therapy. However, data on young adults are largely absent from the literature. This pilot study investigated the effectiveness of wilderness therapy for 297 young adult participants. The 3-year study found clinically and statistically significant change from intake to discharge on the Outcome Questionnaire-45.2. It also found significant change on measures of life effectiveness, motivation for therapy, therapeutic alliance, and dysfunctional attitudes. This article discusses when change occurs in the wilderness, factors that influence outcomes, differences between genders, and post-discharge outcomes for young adults.

Adolescent developments in limbic structures and the endogenous cannabinoid system suggest that teenagers may be more vulnerable to the negative consequences of marijuana use. This study examined the relationships between amygdala volume and internalizing symptoms in teenaged chronic marijuana users. Participants were 35 marijuana users and 47 controls ages 16–19 years. Exclusions included psychiatric (e.g., mood and anxiety) or neurologic disorders. Substance use, internalizing (anxiety/depression) symptoms and brain scans were collected after 28 days of monitored abstinence. Reliable raters manually traced amygdala and intracranial volumes on high-resolution magnetic resonance images. Female marijuana users had larger right amygdala volumes and more internalizing symptoms than female controls, after covarying head size, alcohol, nicotine and other substance use (p

The objective of this study was to investigate the expression of cyclooxygenase (COX)-2 in human choroidal neovascular membranes.Paraffin-embedded sections of choroidal neovascular membranes excised from 16 patients with wet age-related macular degeneration were used for this study. Sections were subjected to immunohistochemistry using a monoclonal mouse antihuman COX-2 antibody. Staining was classified as either negative or positive in retinal pigment epithelial cells, vascular endothelial cells, and fibroblasts. Serial sections were stained for vimentin expression to confirm tissue antigenicity.Eleven of 16 (69%) choroidal neovascular membranes stained positive for COX-2 in retinal pigment epithelial cells, with 6 (38%) of these also expressing COX-2 in vascular endothelial cells and 6 (38%) in fibroblasts. None of the sections that were negative for COX-2 in the retinal pigment epithelial cells showed COX-2 expression in the other cell types assessed. There was a statistically significant difference (P = 0.0097) in the mean ages between the COX-2 positive group (65.6 years) and COX-2 negative group (76.8 years) as determined by a two-tailed, unpaired Student's t-test.The expression of COX-2 in human choroidal neovascular membranes suggests a possible role for this modulator in age-related macular degeneration pathogenesis. The age-dependent expression observed is novel and warrants further investigation.

Background Ocular melanoma is easily treated by the removal of the eye or through plaque radiotherapy. However, after removal or control of the primary tumor, patients can develop fatal liver metastases up to 20 years later. It has been reported that difficulties in imaging single cells and the propensity for tumor cells to replicate rapidly in animal models account for the deficit of single-cell tumor dormancy models. Methods In this paper, we performed two animal experiments using green fluorescent-labeled uveal melanoma cells in nude mice. Cells were injected via tail-vein and the experiments ran 20 and 42 days, respectively. Labeled cells were imaged in vivo via skin-flap and epifluorescent microscopy. Results The first experiment exemplified the feasibility of a single-cell tumor dormancy model; cells were present in multiple organs post-injection, but persisted solely in the liver for the duration of the experiment. The second experiment, demonstrating the presence and viability of these single, metastatic seeds 6 weeks after injection. Conclusion Due to the inherent difficulties in establishing single-celled tumor dormancy models, few exist. In this paper, we have successfully developed a single-cell dormancy model of uveal melanoma, a disease that, in patients, epitomizes tumor dormancy. This model has the potential to reveal the mechanisms behind dormancy, identify patients at high risk for metastatic development, and develop new serum biomarkers for micrometastasis detection.

To evaluate the proliferation rates of five human uveal melanoma (UM) cell lines after treatment with amfenac, a cyclooxygenase (COX)-2 inhibitor, and subsequent radiation exposure.Five human UM cell lines (92.1, SP6.5, MKT-BR, OCM-1, and UW-1) and one human fibroblast cell line (BJ) were incubated with amfenac. Treated and non-treated cell lines were then exposed to various doses of gammaradiation: 0, 2, 4, 6, and 8 Gy. Sulphorhodamine-B assay was used to assess proliferation rates 48 h post-radiation.Treatment of UM cell lines with amfenac prior to radiation led to a marked reduction in proliferation rates. This difference was statistically significant in all cell lines at every radiation dose (P0.005), with the exception of 92.1 at 2 Gy (P=0.157). Fibroblasts treated with amfenac showed significantly higher proliferation rates after 2 and 8 Gy, with no significant differences at 0, 4, and 6 Gy.The radiosensitivity of UM cell lines was increased by the administration of amfenac, the active metabolite of nepafenac. There appears to be a radioprotective effect of amfenac on human fibroblasts. The topical administration of nepafenac may decrease tumour recurrence and radiation-induced complications while broadening the indications for radiotherapy by treating larger tumours.

Uveal melanoma (UM) is the most common malignant intraocular tumor in adults. Despite the high accuracy of clinical diagnosis and advances in local treatment, more than 50% of UM patients develop metastasis within 10 years of initial diagnosis. NM23 is one of the human metastasis suppressor genes. Reduced nm23-H1 expression is correlated with high metastatic potential in many different cancers. The purpose of this study is to investigate the expression of nm23-H1 in UM and its potential value as a prognostic marker. Immunostaining of nm23-H1 was verified in five human UM cell lines with different metastatic potentials. The expression level of nm23-H1 mRNA was evaluated with one-step quantitative real-time PCR. The invasion ability of the cell lines was assessed before and after silencing nm23-H1 with small interference RNA. Thirty-two cases of paraffin-embedded specimens of human UM were immunostained with nm23-H1 monoclonal antibody. The immunostaining was evaluated in a semiquantitative fashion based on extent and intensity. The real-time PCR results of five human UM cell lines showed that expression of nm23-H1 was higher in cell lines with low metastatic potential compared with those with high metastatic potential (P

Objective: It was the aim of this study to determine the diagnostic accuracy of high-risk prognostic factors and morphological characteristics of retinoblastomas using digital whole slide images (WSI) generated by a scanner. Methods: Forty-seven H&E sections of glass slides with high-risk morphological features of retinoblastoma were analyzed. Slides were scanned as WSI and reviewed. The results were compared with those obtained after reviewing the slides using a regular microscope as the gold standard. McNemar's test (MT), the percentage of agreement (POA), and sensitivity (S) and specificity (Sp) were evaluated between WSI and conventional microscopy. Results: There were no differences with respect to multicentricity, growth type, rosette formation, choroidal invasion, anterior chamber invasion, extraocular extension, scleral extension, optic nerve invasion, necrosis, or Azzopardi effect between WSI analysis and light microscopy (MT, p = 1.0; POA = 100%; S = 100%, and Sp = 100%). Discordance was found in 1 case where calcification could not be found using WSI (MT, p = 1.00; POA = 97.9%; S = 100%, and Sp = 97.8%). Conclusion: To the best of our knowledge, this is the first report using digital pathology (WSI) to evaluate prognostic factors in eyes containing retinoblastomas. Using WSI, the pathologist was able to detect high-risk morphological features in retinoblastoma. To date, WSI is an important tool, in particular for ophthalmic pathologists examining enucleation and exenteration specimens.

Background: Despite the fact that retinoblastoma treatment has dramatically increased the survival and vision preservation in these patients, it is still important to pursue new therapeutic targets to minimize the side-effects of current therapies. Gonadotropin releasing hormone (GnRH) has been shown to exert a direct anti-proliferative effect on many types of malignancies, such as breast cancer, skin melanoma, and glioblastoma. The aim of this study is to describe the presence of GnRH receptor (GnRHR) in retinoblastoma in order to identify a possible new therapeutic target for this disease. Methods: Protein expression of GnRHR was studied by immunohistochemistry in 32 eyes with retinoblastoma and in the Y79 retinoblastoma cell line. Expression was scored according to intensity (1-3) and distribution (1-4), which were multiplied to generate an immunoreactive score (IRS). A score of 1 to 6 was considered low, while scores higher than 6 were considered high. The Fisher’s exact test was used to compare GnRHR IRS with presence or absence of each morphological high-risk feature. Results: GnRHR was expressed in all retinoblastoma cases and in the Y79 cell line. High, and low expression according to IRS score was evident in 62.5% and 37.5% of cases. There were no differences in GnRH IRS with respect to uni- versus multi-focal tumors, type of growth (mixed/endophytic), rosette formation, choroidal invasion, extraocular extension orextension to the sclera, or optic nerve invasion. Conclusion: GnRHR is expressed in varying degrees in all retinoblastomas. Therefore, GnRHR may be a novel therapeutic target for the treatment of retinoblastoma. Moreover, GnRHR expression could be a biomarker for the responders. Nevertheless, GnRHR expression did not correlate with morphological prognostic factors in this series. Further studies with a larger number of cases and follow up are needed to confirm our results.

Uveal melanoma is the most common malignancy of the adult eye. Although it is a relatively infrequent tumor, clinical prognosis is often poor owing to a high incidence of aggressive metastatic disease, for which there are limited treatment options. Little is known about the etiology of this condition, although several risk factors have been identified. Unlike cutaneous melanoma, however, ultraviolet radiation does not figure prominently among these risk factors. In this review, we focus on an associated form of visible electromagnetic radiation, high-energy short-wave (blue) light, a causative agent in various forms of age-related retina damage, as a previously overlooked risk factor in uveal melanoma development and progression. Finally, we discuss the impact of these data on contemporary ocular therapy, particularly the debate surrounding the filtering capabilities of intraocular lenses used to replace dysfunctional crystalline lenses during cataract surgery.

When performing a modal test, a number of mechanisms are available to approximate the free-free boundary condition. Some test setups may be more difficult to implement than others, but ultimately the effect of the boundary condition on the actual modes of the system are of concern. An expensive setup may not be any better than an equivalent, inexpensive “Rube Goldberg” approach. In lieu of an expensive setup, a low cost, easily deployed configuration may be improvised from common objects not native to the test lab.

Introduction: Superficial epithelioma with sebaceous differentiation is a rare benign epithelial neoplasm. It usually involves the head, neck or the back of a middle-age person. To the best of our knowledge, two ocular cases have been reported in the literature. Case presentation: A 46-year-old man of Italian descent, with a known history of testicular seminoma treated by orchiectomy with chemotherapy and radiotherapy, presented with a tan-colored lesion measuring 4mm in diameter in his right upper lid that had been growing over 10 months. It was clinically diagnosed as papilloma. An excisional biopsy was done. On histological examination, the lesion was a well-circumscribed and sharply demarcated epithelial tumor attached to the overlying epidermis and characterized by plate-like proliferation of basaloid to squamous cells with clusters of mature sebaceous cells and foci of ductal differentiation. After a follow-up period of 5 months, no recurrence of the lesion has been documented. Conclusions: Superficial epithelioma with sebaceous differentiation is part of the differential diagnoses of eyelid lesions. Arguments in the literature about the correct nomenclature of superficial epithelioma with sebaceous differentiation have resulted in under-diagnosed cases. The benign histological features and the lack of recurrence support its benign nature. Although no clear association has linked superficial epithelioma with sebaceous differentiation with Muir–Torre syndrome, further clinical correlation and close follow up for patients are recommended.

Background/aims Sebaceous adenomas (SAs) are rare, benign sebaceous gland tumours of the eyelid. SAs may be associated with primary internal malignancies. This association is known as Muir-Torre Syndrome (MTS). The purpose of this study was to approximate the prevalence of SAs, to determine the reliability of the clinical diagnosis of SAs and to demonstrate immunohistochemical staining of DNA mismatch repair proteins mutL homologue 1 (MLH1) and mutS homologue 2 (MSH2) for a case of MTS. Methods We reviewed the histopathology reports from all eyelid specimens collected between 1993 and 2013 at the Henry C Witelson Ocular Pathology Laboratory to determine the proportion of SAs. For the SAs identified on histopathology, we looked at patient charts to see what diagnosis was originally suspected on clinical examination. Immunohistochemical staining for MLH1 and MSH2 was performed on all SAs to screen for MTS. Results Of the 5884 eyelid specimens collected, 9 were SAs (6 women, 3 men; 42–72 years old). The diagnosis of SA was suspected clinically in only one of the nine cases based on the gross appearance of the eyelid lesion. Immunohistochemistry revealed one SA case with positive MLH1 expression and negative MSH2 expression. These findings prompted systemic work-up and this patient was diagnosed with MTS after discovery of a colon adenocarcinoma T2M0N0. Conclusions The diagnosis of eyelid SA is rare. The importance of this benign eyelid tumour stems from its association with internal malignancies in MTS. Immunohistochemical staining of mismatch repair proteins MLH1 and MSH2 is a valid and accessible strategy for investigating MTS in patients with SAs.

Below the knee amputation after trauma is an appropriate option for many patients with recalcitrant infection and nonunion of the tibia. Patients who have had transtibial amputations have lower energy expenditure, heart rate, and oxygen cost when ambulating with their prostheses than when using a three-point gait with crutches without their artificial limb. Innovative prosthetists have improved each of the five essential components of the limb amputated below the knee: socket, insert, shaft and pylon, foot and ankle assembly, and suspension system. Prosthetists are integral members of the patient's healthcare team. Their recommendations and direct patient care are essential to optimizing the functional ability of patients who have had amputations.

Background: Uveal melanoma (UM) is a disease that affects approximately five people per million in the United States. This disease metastasises predominantly to the liver, and treatment options following the clinical detection of these sequelae are limited. Vascular endothelial growth factor-A (VEGF-A) is the primary activator of tumour angiogenesis and functions by binding to VEGF-Receptor 2 (VEGF-R2) and is often required for tumour growth beyond 2–3 mm. The purpose of this study was to investigate the expression of VEGF-A and the primary VEGF-R2 in three UM cell lines. Furthermore, we investigated the effects of VEGF-A inhibition on receptor activation and production of other cytokines. Finally, the effects of VEGF-A inhibition on the proliferation, migration, and invasion in the cell lines were ascertained. Materials: Three UM cell lines (92.1, OCM-1, and UW-1) were incubated with and without the addition of 100 μg/mL of bevacizumab. VEGF-A expression under both conditions was determined by sandwich enzyme-linked immunosorbent assay (ELISA), and phosphorylated VEGF-R2 expression was determined using western blot. The effects of VEGF-A inhibition on 20 cytokines (IL-1a, IL-2, IL-5, IL-8, IL-12p70, GM-CSF, IFNy, CCL3, MMP-9, TNF-a, IL-1b, IL-4, IL-6, IL-10, IL-13, GRO, MCP-1, MIP-1b, and RANTES) were determined using a multiplex sandwich ELISA. Proliferation rates before and after treatment were evaluated via sulforhodamine B assay, and migration and invasion assays implementing the Boyden chamber technique, the latter with artificial extracellular matrix, were used to assess their respective abilities. The Student’s t-test was used to compare changes in cytokine expression following VEGF-A inhibition. Analysis of variance was used to compare changes in the functional abilities of three uveal melanoma cell lines following VEGF-A inhibition. A P-value < 0.05 was considered statistically significant. Results: All three cell lines produced copious amounts of VEGF-A in culture (92.1, 11785.5 ± 231.8 pg/μL; OCM-1, 4608.0 ± 324.0 pg/μL; UW-1, 8309.3 ± 634.5 pg/μL), which was reduced to undetectable levels following the administration of bevacizumab (P< 0.05). Similarly, detectable phosphorylated VEGF-R2 was present in all cells, which was reduced significantly in all cell lines following bevacizumab treatment (107525.2 ± 8602.0 versus 1024.5 ± 98.2, 46587.3 ± 4192.9 versus 12821.1 ± 1666.7, and 60394.3 ± 4026.4 versus 6908.2 ± 607.2; 92.1, OCM-1, and UW-1, respectively; P< 0.05). Of the cytokines investigated, only MMP-9 and CCL3 were ubiquitously altered across all three cell lines following bevacizumab treatment; they were upregulated (CCL3: 1072.50 ± 18.77 pg/mL versus 1281.00 ± 72.34 pg/mL; 22.5 ± 7.85 pg/mL versus 62.00 ± 9.16 pg/mL; 20.33 ± 6.35 pg/mL versus 35.00 ± 6.22 pg/mL; control versus bevacizumab; MMP-9: 25.50 ± 5.47 pg/mL versus 88.25 ± 13.38 pg/mL; 19.75 ± 4.14 pg/mL versus 45.25 ± 8.36 pg/mL; 3.25 ± 1.09 pg/mL versus 19.25 ± 3.77 pg/mL; control versus bevacizumab; 92.1, OCM-1, and UW-1, respectively; P< 0.05). Bevacizumab significantly reduced the proliferation of one cell line (92.1: 0.405 ± 0.012 versus 0.509 ± 0.033; bevacizumab versus control; values OD; P< 0.05), the migration of two cell lines (92.1: 0.071 ± 0.003 versus 0.115 ± 0.003; OCM-1: 0.049 ± 0.005 versus 0.117 ± 0.014; bevacizumab versus control; values OD; P< 0.05), and did not significantly affect invasion. Conclusion: Despite the significant reduction in phosphorylated VEGF-R2 levels, bevacizumab did not have a dramatic impact on the functional abilities of the three UM cell lines studied. Our results indicate that compensatory mechanisms, such as the upregulation of MMP-9 and CCL-3, following bevacizumab administration may mitigate its effects on these abilities.

In recent years, the advent of new imaging technologies has enabled researchers to portray the development of metastases in animal models in a new light. Previously, observing the behaviour of metastatic cells required the sacrifice of the animal and thus the progress of development or treatment is often incomplete and necessarily elucidated from sequential sacrifices of inbred animals. Considering the technical difficulties in visualizing individual metastatic cells by traditional immunohistochemical or staining methods, understanding the critical first steps of micrometastatic development was difficult and often impractical. New technologies, in particular the ability to force cells of interest to fluoresce amongst a dull background, have enabled researchers to visualize the behaviour of individual metastatic cells in host organisms. These methods, in conjunction with new, non-lethal intravital imaging methods, have contributed greatly to the body of knowledge regarding malignant cells and the development of metastases. This chapter will discuss several of the more popular imaging methods such as luciferase, green fluorescent protein, MRI, CT, PET, and will analyze the advantages and limitations of each method.

Spontaneous, chemically induced, and transgenic models of metastases provide a unique framework for studying disease progression; in most cases, they allow the observation of the full spectrum of metastasic development including intravasation, survival in the blood, extravasation, organ seeding, and proliferation, in one immunocompetent animal. This is an advantage over haematogenous and ectopic counterparts that only offer the ability to observe specific fragments of the metastatic cascade. However, observing the intricate processes of metastasis, in particular those such as extravasation and intravasation, which involve single cells, in a spontaneous or induced model is challenging. With respect to chemically and radiation induced models, the methods needed to initiate tumor formation poses potential health risks to the practitioner. Spontaneous and induced models also do not permit the study of human malignant cells; due to the nature of these models, observations made regarding the behaviour and development of metastasis must be prefaced by mentioning that these cells are of animal origin. However, spontaneous, induced, and transgenic models offer a unique background for the testing of pharmaceutical compounds in an immunocompetent animal that represents all stages of malignant development.

Animal models of cancer and of metastasis in particular, provide a critical link between in vitro studies and the treatment of human disease. Animal models allow scientists to understand and interpret disease pathogenesis in an environment in which metastatic cells are constantly bombarded by autocrine, paracrine, and endocrine signals from a multitude of sources. Disease pathogenesis can only be elucidated through observation of organs, organ systems, and, ultimately, whole organisms. In addition to contributing to our knowledge of disease pathogenesis, animal models also provide an avenue for testing both the safety and efficacy of various anti-metastatic drugs and biological compounds. Animal models allow interpretation of the physiological effects of administrating a drug or biological compound in a complex organism. In this respect, animal models are critical to the drug development process and are a required element of the drug approval process. Within the umbrella term ‘animal models’, there are various types of models that present unique advantages and disadvantages to understanding metastasis and the ability to test anti-metastatic compounds. Ideally, animal models that closely mimic human cancers, with respect to disease duration, progression, mode of dissemination, and metastatic location, are desirable. However, due to the often inefficiency that is inherent in the metastatic process, animal models in which metastasis takes many years to develop are impractical. To overcome this innate obstacle, several different types of animal models have been developed to expedite the metastatic process: there are ectopic models, in which malignant cells are explanted into foreign locations on the animal, and there are haematogenous models of metastasis, in which cells are dispersed into circulation with the intention of simulating a natural course of dissemination.

Secreted protein acidic and rich in cysteine (SPARC) has been shown to play an integral role in the progression of numerous malignancies. The aim of this study was to investigate the expression of SPARC in uveal melanoma (UM).SPARC expression was assessed in UM cell lines using RT-PCR and immunocytochemistry. Small interfering RNA directed against SPARC was used to transfect each of the cell lines, which were subsequently run in proliferation assays. SPARC expression was further investigated in 19 cases of human UM and 11 primary and 8 metastatic tumors from a rabbit xenograft model.The cell lines transfected with SPARC siRNA showed a significant decrease in proliferation compared to controls. All cases of human uveal melanoma demonstrated positive staining for SPARC as did all primary and metastatic tumors from the xenograft model.SPARC may represent a novel target to inhibit growth of UM.

Summary Aims Keratoconus is a bilateral and asymmetrical corneal ectasia. The pathophysiology of this disorder has yet to be fully elucidated. The purpose of our study was to document the prevalence of the most common morphological features of keratoconic corneas. Methods A retrospective analysis of 49 cases diagnosed as keratoconus between 2001 and 2006 was undertaken. Histopathological reports were reviewed to obtain data such as age and gender. Specimens were fixed in 10% buffered paraformaldehyde solution for 24 h, bisected through the centre of the button, and embedded in paraffin. Sections were stained with haematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) for light microscopic examination. Results The studied group was composed of 29 men and 20 women. Age at the time of the penetrating keratoplasty was 39±14 years (mean ± standard deviation). Forty of the 49 specimens (82%) presented with epithelial thinning. Other common features of keratoconus included breaks in Bowman’s layer in 35 (71%), compaction of the stromal collagen fibres in 31 (63%), and folds in Descemet’s membrane in 31 (63%) cases. Other less common histopathological findings were: presence of superficial iron deposits in 14 (29%), deep stromal scarring in 12 (24%), epithelial scarring in 11 (22%), endothelial cell loss in 11 (22%), and breaks in Descemet’s membrane in nine (18%) cases. Conclusions Some of the histopathological findings associated with keratoconus are subtle. It is important to be aware of them in order to properly confirm the clinical diagnosis.

Uveal melanoma arises from melanocytes located in the uveal tract of the eye and is the most common primary intraocular tumor in adults. Metastatic liver disease is the overwhelming cause of death in uveal melanoma patients, with almost 50% of patients developing liver metastases up to 15 years after diagnosis. Most of these patients do not present with any evidence of overt metastasis at the time of initial diagnosis although it is assumed that they have undetectable micrometastases. Currently, there are no therapeutic modalities to prevent or efficiently treat the metastatic disease in uveal melanoma patients. Recent discoveries have shed light on the molecular pathways that may contribute to the progression of liver metastasis. The aim of this review is to describe new insights into the genetic and molecular pathways that may play a role in the development of liver metastases in uveal melanoma patients.

Purpose: We previouly demonstrated the feasibility of a human uveal melanoma GFP model in nude mice. We were able to monitor the preferential colonization of the liver by malignant cells during a 20-day period. In this study we extended the observation period in order to observe disease progression and metastatic growth. Methods: Five hundred thousand, GFP transfected, human primary uveal melanoma cells (92.1) were injected into the tail vein of 30 nude mice at time zero. Starting at day one, five mouse per week was imaged, using an abdominal incision to expose the liver, for up to 60 minutes before sacrifice. The liver was imaged in vivo and post mortem, while the lungs, spleen, and kidney were all examined post-mortem for GFP expression. Histopathological examination of all organs was performed to ensure that the GFP signal was arising from malignant cells. Results: In vivo imaging of the liver revealed positive GFP signal in all mice throughout the experiment. Malignant cells seeded the liver after injection, but there was no apparent tumor growth upon completion of the experiment. The micrometastatic foci remained viable without changes in size or intensity of the signal. No tumors were seen in any other organ at the end of the experiment. Histopathology confirmed that the cells emitting GFP were malignant cells. Conclusions: The results of this uveal melanoma model show great promise for understanding the previously unobservable interactions between single human uveal melanoma cells and native liver tissue. The fact that our cells successfully colonize the liver yet remain dormant for six weeks should actually mirror the disease in humans; metastases in patients takes roughly five to ten years to develop.

PURPOSE To determine the frequency and describe the histopathologic features of stromal corneal dystrophies over a 10-year period. METHODS A single-centered, retrospective analysis was performed on corneal specimens diagnosed as stromal dystrophies retrieved from the Henry C. Witelson Ophthalmic Pathology Laboratory and Registry (Montreal, Canada) over a period of 10 years. Corneal specimens obtained during keratoplasty were subjected to hematoxylin and eosin, periodic acid-Schiff, and other special staining. RESULTS During the period of the study, 885 corneal specimens from penetrating keratoplasties were received. Twenty-six (2.9%) specimens from a total of 22 patients showed stromal corneal dystrophies: 17 (65.4%) eyes with lattice dystrophy, 5 (19.2%) eyes with combined granular/lattice ("Avellino") dystrophy, 3 (11.5%) eyes with granular dystrophy, and 1 (3.9%) eye with macular dystrophy. CONCLUSIONS Corneal stromal dystrophies have distinct histopathologic features, which can be diagnosed with routine and special staining, particularly for combined granular/lattice (Avellino) dystrophy. Clinically, combined granular/lattice (Avellino) dystrophy can have features of both lattice and granular dystrophies, which can be misleading for diagnosis. Lattice dystrophy was the most frequent stromal dystrophy in this particular population.

Herein, a case of an unusual ocular presentation of primary antiphospholipid syndrome. The patient disclosed localized retinal ischemia due to branch retinal artery occlusion, confirmed by fluorescein angiography and manual perimetry.The ocular manifestation led to the diagnosis and successful treatment of the systemic disease.

The expression of cyclooxygenase-2 (COX-2) has been reported as an indicator of poor prognosis in a wide variety of human tumors, including colon, breast and uveal melanoma (UM). COX-2 inhibitors have shown promise in controlling the malignancy of several types of tumors. Previous studies have demonstrated the efficacy of a COX-2 inhibitor on the proliferation rates of human UM cells. The goal of this experiment was to investigate the efficiency of Nepafenac, a topically administered COX-2 inhibitor, in a rabbit model of UM. The animals were divided into two groups of 14 animals for the duration of the 12-week experiment. One animal per group was killed each week to evaluate disease progression and for histopathological studies. The experimental group received drops containing 0.3% Nepafenac solution. Intraocular tumor growth was evaluated weekly by fundoscopic examination and each animal was weighed prior to examination. Blood samples were taken weekly from all rabbits to detect circulating malignant cells (CMCs) throughout the experiment. After the second week of inoculation, the experimental group weighed significantly more than the control group. The control group developed more intraocular tumors and presented with metastases and higher detectable levels of CMCs before the treated group. These results indicate that the topical administration of a COX-2 inhibitor delayed the progression of this malignancy in our animal model. A clinical trail using an anti-COX-2 inhibitor for patients with UM should be considered.

Background Melan-A and tyrosinase are new immunohistochemical markers that can be used in the diagnosis of melanocytic lesions. The aim of this study was to investigate the correlation between radiotherapy or clinicohistopathological parameters and the expression of melan-A and tyrosinase in uveal melanoma. Methods Thirty-six enucleated cases of uveal melanoma were studied. The formalin-fixed, paraffin-embedded specimens were immunostained with monoclonal antibodies against melan-A and tyrosinase. The samples were classified as either positive or negative. The chi-square or the Student-t tests were used to test for the correlation of the expression rates of melan-A and tyrosinase with clinico-pathological parameters. Results Melan-A and tyrosinase were positive in 33 (91.7%) and 35 (97.2%) of the specimens, respectively. There was no significant association between the expression of melan-A or tyrosinase and radiotherapy or any clinico-pathological parameter. All specimens were positive for at least one of the immunohistochemical markers. Conclusion To the best of our knowledge this is the first study concluding that the expression of melanocytic markers such as melan-A and tyrosinase is not influenced by radiotherapy or any clinico-pathological parameter. Moreover, when tyrosinase and melan-A are used together, 100% of the formalin-fixed, paraffin-embedded uveal melanoma samples tested positive for one of those markers.

The influence of simple carboxylates on oilfield chemistry measurements, steel corrosion, and corrosion inhibition was studied. In most cases, the error in the field measurement of dissolved CO2 was found to be small. In all but two cases examined, the error in bicarbonate ion measurement was small because of the large actual bicarbonate ion to carboxylate ratio. The presence of acetate and propionate ions in laboratory tests always accelerated the uninhibited corrosion rate of mild steel because of an increase in cathodic kinetics when the steel was behaving actively. In passive systems, anodic increases were seen. Both acetate and propionate were detected in field samples; in only one case butyrate was observed. Corrosion inhibitors were effective at controlling corrosion even in the presence of these ions in both laboratory and field conditions.

Frequently, patients undergoing complex bone reconstructions for the treatment of osteomyelitis complicating tibial nonunion require extended exoskeletal support. Once the need for strict, continuous cast immobilization has passed, the patient can be fitted with a removable orthosis. Although prefabricated ankle foot orthoses are available, the purpose of this paper is to highlight the sophisticated new custom orthotic options available to the physician for the patient. Difficult to fit limbs now can be accommodated. Access to a skilled orthotist is the sine qua non of success.