Toxicity of the photosensitizer NPe6 following intravitreal injection.

BACKGROUND AND OBJECTIVE: To determine the retinal toxicity of mono-L-aspartyl chlorin e6 (NPe6) following intravitreal injection. METHODS: Twelve Dutch-belted rabbits divided into 5 experimental groups (n=2 each) were injected intravitreally with 6.25, 12.5, 25, 50, or 100 microg of NPe6; one control group (n=2) was injected with intravitreal normal saline. One eye in each rabbit was sutured shut to test the effect of light exposure. Fundus photography and electroretinograms were performed before treatment and 2 days, 1 week, and 2 weeks after injection. Animals were euthanized and the eyes enucleated for histopathologic analysis. RESULTS: After 1 week, 4 uncovered eyes given 50 and 100 microg had central retinal vein occlusion and varying degrees of retinal hemorrhage. RPE proliferation was seen in the covered eyes given 50 or 100 microg. Electroretinograms revealed absent retinal response at 100 microg and mild toxicity at 50 microg, but no change from normal at doses of < or = 25 microg of NPe6. CONCLUSIONS: Intravitreal doses of < or = 25 microg NPe6 caused little or no apparent toxicity; however, toxicity was significant at doses of 50 microg and 100 microg.

Mardi Gras eye injury survey, 1998-1999.

BACKGROUND: We studied the nature of associated ocular trauma during the 1998 and 1999 New Orleans parade seasons and whether trends were observable from previous surveys. METHODS: A prospective survey and retrospective analysis included 18 emergency rooms (ERs) in the New Orleans area. RESULTS: Sixteen surveys representing eight ERs were completed between February 19 and 24, 1998, and 32 surveys representing five ERs were completed between February 5 and 17, 1999. In both years, the most common ocular complaints were pain, blur, foreign body sensation, tearing, and photophobia. The most common slit lamp findings in 1998 were within normal limits, corneal abrasion, and conjunctival hyperemia. In 1999, the most common findings were subconjunctival hemorrhage, corneal abrasion, cell and flare, and lid laceration. CONCLUSIONS: Projectile injuries of the eyes are common during Mardi Gras due to the nature of interaction between paradegoers and float riders. We found no identifiable trend in the number or type of injuries reported in 1986, 1987, and 1998 Mardi Gras surveys.

The relationship between human papillomavirus and p53 gene in conjunctival squamous cell carcinoma.

PURPOSE: The p53 tumor-suppressor gene has been documented to exist in mutated forms in many types of squamous cell carcinoma in the body. Also in conjunctival squamous cell carcinoma, human papillomavirus (HPV) is accepted as an oncogenic factor. The objective of our study was to establish a correlation between p53 overexpression and the presence of HPV infection within tumor tissues from patients with conjunctival squamous cell carcinoma. METHODS: Tissue sections obtained from paraffin-embedded conjunctival squamous cell carcinoma specimens from 23 patients were examined with light microscopy, polymerase chain reaction (PCR), and immunohistochemistry. RESULTS: Seventy-eight percent of tumors were positive for p53, whereas 22% were positive for HPV. The proportion of patients positive for both p53 and HPV was 17%, whereas another 17% of the patients were negative for both p53 and HPV. Therefore no significant disproportion was found in the distribution of patients' HPV status and p53 status (p = 1.00). No significant correlation or linear association was found between the HPV status and p53 status (r = 0.022; p = 0.920). CONCLUSION: We could not show any statistical association between abnormal p53 gene-product expression by immunohistochemistry in conjunctival squamous cell carcinomas and HPV infection by PCR detection techniques.

Threshold power levels for NPe6 photodynamic therapy.

OBJECTIVE: To determine the threshold power levels for producing retinal and choroidal vascular occlusion using mono-L-aspartyl chlorin e6 (NPe6) photodynamic therapy; to evaluate its efficacy with longer intervals between photosensitizer injection and laser application; to determine the elapsed time between light application and appearance of angiographic changes. METHODS: Pigmented and nonpigmented rabbits were injected intravenously with 2 mg/kg of NPe6 before laser irradiation of the retina-choroid. Group 1 was treated at increasing power levels; fluorescein angiograms were obtained at each fluence. Group 2 animals were exposed to laser irradiation at 5 minutes, and 1 and 3 hours postinjection to determine (by fluorescein angiography 24 hours post-treatment) if increasing the interval affected outcome. Group 3 animals underwent fluorescein angiography at 30 minutes, 1 hour, 2 hours, and 24 hours posttreatment to document the time between laser application and subsequent vessel closure. RESULTS: Choroidal vessel occlusion was angiographically evident in all lesions at fluences of > or = 2.65 J/cm2 in pigmented rabbits and at > or = 0.88 J/cm2 in nonpigmented rabbits. Lesion diameter decreased as the time between injection and treatment increased. Vessel occlusion was documented at least 2 hours after treatment. CONCLUSION: Choroidal vessel occlusion can occur at very low fluence.

Enucleation.

The three most common indications for enucleation are intraocular malignancy, trauma, and a blind, painful eye. Recommending enucleation is one of the most difficult therapeutic decisions in ophthalmology. In some cases of malignancy, cryotherapy, laser photocoagulation, diathermy, chemotherapy, and radiation therapy may be viable alternatives to surgery. When surgery is chosen, evisceration or exenteration may be alternatives to enucleation. Once the decision is made to perform enucleation or evisceration, the surgeon must choose from several types of implants and wrapping materials. These devices can be synthetic, autologous, or eye-banked tissues. With certain implants, the surgeon must decide when and if to drill for subsequent peg placement. In this review, the authors discuss choices, techniques, complications, and patient consent and follow-up before, during, and after enucleation. Controversies and results of the Controlled Ocular Melanoma Study are summarized.

Ocular vascular thrombosis following tin ethyl etiopurpurin (SnET2) photodynamic therapy: time dependencies.

BACKGROUND AND OBJECTIVES: To evaluate the optimal time from the end of photosensitizer injection to the commencement of light application for creating characteristic fundus lesions and the time to vascular occlusion following photodynamic therapy (PDT) with tin ethyl etiopurpurin (SnET2). MATERIALS AND METHODS: Following intravenous injection of SnET2 0.5 mg/kg or lipid emulsion alone, the fundus of rabbits was irradiated at different times (5 to 240 minutes) after photosensitizer injection using 664 +/- 7-nm laser light with an irradiance of 354 mW/cm2 and fluence of 20 J/cm2. Ophthalmoscopy and fluorescein angiography were performed 1 day after SnET2 PDT. In separate groups of rabbits, treated areas of the fundus were imaged within 30 minutes following PDT using fluorescein vesicle and microsphere angiography with scanning laser ophthalmoscopy to document time of vascular occlusion. All animals were killed 1 day following treatment and eyes were examined by histopathology. RESULTS: Areas of hypofluorescence (indicating vascular occlusion) were seen when activating laser light was applied 5 to 20 minutes after SnET2 injection. Retinal vessels remained perfused in all cases. The time to vascular occlusion was 70 to 120 and 40 to 90 minutes in nonpigmented and pigmented rabbits, respectively. No safety issues were seen. CONCLUSION: PDT with SnET2 was effective in occluding the choriocapillaris. Activating light needs to be applied within a specific time frame after photosensitizer injection to achieve vascular occlusion.

Ischemia induces significant changes in purine nucleoside concentration in the retina-choroid in rats.

Adenosine, produced from the decomposition of adenosine triphosphate, is believed to provide protective effects during ischemia. On the other hand, adenosine metabolites may serve as precursors for oxygen free radical formation. The time course of formation of adenosine and its purine metabolites was studied during retinal ischemia in rats. Concentrations of adenosine and its purine nucleoside metabolites inosine, hypoxanthine, and xanthine in the retina-choroid of ketamine/xylazine-anesthetized rats were measured during retinal ischemia using high performance liquid chromatography. Quantitative measurements were made possible in the small tissue mass through the use of internal standards. Ischemia was induced by ligation of the central retinal artery. In each rat, one eye was ischemic while the other served as a non-ischemic control. Eyes were frozen in situ at 1, 5, 10, 20, 30, 60, and 120 min of ischemia. The retina-choroid was then removed from the frozen eyes and analysed. Significant increases in the concentrations of adenosine, inosine, and hypoxanthine in ischemic compared to control retina-choroid were detectable within 1 to 5 min of the onset of ischemia, and within 10 min for xanthine. Increase in adenosine concentration in ischemic relative to control retina-choroid plateaued at 30 min of ischemia, while inosine and hypoxanthine concentrations increased continuously. The increase in xanthine concentration was exponential throughout the measurement period. This study documented the time-related changes in purine nucleoside concentration during ischemia. Prolonged ischemia results in ongoing production of xanthine, which by serving as a precursor for oxygen free radical formation, could be a pathogenic factor in prolonged retinal ischemia.

Measurement of purine nucleoside concentration in the intact rat retina.

Adenosine, produced from the decomposition of adenosine triphosphate, is believed to provide protective effects during ischemia. On the other hand, adenosine metabolites may serve as precursors for oxygen free radical formation. These substances have not been previously measured in intact vertebrate retina, where adenosine and its metabolites may play a role in the pathogenesis of ischemic injury. The small tissue mass of the retina, particularly in rats, renders these measurements challenging. Furthermore, accurate measurement of purine nucleosides requires immediate cessation of ongoing adenosine metabolism. Concentrations of adenosine and its purine nucleoside metabolites inosine, hypoxanthine, and xanthine in the retina of ketamine/xylazine-anesthetized rats were measured after in situ freezing using high-performance liquid chromatography. The retina was removed from the frozen eyes and analyzed. Quantitative measurements were made possible through the use of an internal standard. Ischemia was induced by ligation of the central retinal artery. Retinal purine nucleoside concentrations did not differ between the two eyes of the rat under control conditions, and there was no effect of placement of the ligating suture itself compared to completely unmanipulated eyes. Use of two different in situ freezing methods yielded comparable results. To evaluate the impact of a period of ischemia, one retina of each rat was ischemic for 30 min, and the other, non-ischemic. Our measurements were associated with a high degree of reproducibility and minimal variability, and significant changes in purine nucleoside concentrations were detectable in the retina after 30 min of ischemia. Our method may be used to assess the role of adenosine and its metabolites in the pathogenesis of ischemic neuronal injury, including in the retina.

Fluorescent vesicle angiography with sodium fluorescein and indocyanine green.

BACKGROUND AND OBJECTIVE: The authors evaluated the feasibility of merging free-dye angiography and the fluorescent vesicle technique to achieve the best characteristics of both. MATERIALS AND METHODS: Fluorescent vesicles encapsulated with either indocyanine green or carboxyfluorescein were mixed with free indocyanine green or free sodium fluorescein, respectively, and imaged with a scanning laser ophthalmoscope in both an in vitro model and primate and rabbit models. RESULTS: In the in vitro model of the sodium fluorescein combination, optimal viewing of vesicleen capsulated dye and free dye was at a ratio of 150:1; for indocyanine green, the ratio was 50:1. In vivo, high-quality fluorescent vesicle angiograms were obtained that demonstrated leakage of free dye from choroidal laser spots. CONCLUSIONS: Free dye and fluorescent vesicles can be combined to obtain an angiogram with all of the advantages of a traditional angiogram, while allowing the operator to assess the changes in retinal or choroidal circulation directly.