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Expression of the c-kit receptor in choroidal melanomas.

The KIT gene encodes c-kit, a transmembrane receptor that has tyrosine kinase activity and plays a role in haematopoiesis, gametogenesis and melanogenesis. The c-kit protein is found in normal cutaneous and choroidal melanocytes, and there is evidence that expression is lost in melanoma. Expression of c-kit was analysed in 57 paraffin-embedded sections of choroidal melanoma specimens and three choroidal melanoma cell lines using immunochemistry and Western blotting. Of the tumour specimens, 75% stained positively for c-kit with a membrane pattern of reactivity. Of the six patients who underwent proton beam therapy before enucleation, five tumours exhibited no c-kit immunoreactivity and the other tumour demonstrated weak staining. Of the three melanoma cell lines used, c-kit expression was observed in only one. No correlations between c-kit positivity and parameters such as cell type, largest macroscopic tumour dimension, scleral invasion or pigmentation were observed. In contrast, a significant positive association was found between c-kit staining and mitotic activity (P = 0.02). However, c-kit expression did not significantly influence survival when evaluated by univariate analysis. In conclusion, c-kit is expressed in most choroidal melanoma tumours. Further analysis should provide new insights into the mechanisms underlying the molecular and cellular changes in choroidal melanomas.

Blotting, Western↗

Present indications for removal of choroidal neovascular membranes.

Subretinal choroidal neovascularization is an important cause of visual loss in adults. Removal of choroidal neovascular membranes using relatively new techniques of subretinal surgery holds promise for improving or stabilizing vision in some patients. A number of factors have been proposed that may affect the outcome of subretinal surgery for choroidal neovascularization, including the degree of involvement of Bruch's membrane, the location of the neovascular complex relative to the retinal pigment epithelium and Bruch's membrane, the location of the ingrowth site, and the etiology of the membranes. Although the surgical techniques are relatively refined, the indications for removal of choroidal neovascular membranes are still under investigation. This article reviews the recent and past literature and summarizes the authors' specific recommendations regarding present indications for removal of choroidal neovascular membranes.

Choroid↗

New treatments for serpiginous choroiditis.

Serpiginous choroiditis is a bilateral, inflammatory condition of the inner choroid and overlying retinal pigment epithelium with a distinctive retinal distribution. Although rare, serpiginous choroiditis deserves attention because it can lead to permanent loss of vision due to foveal involvement by inflammation or choroidal neovascularization from frequent recurrences. This article reviews the recent ophthalmic literature published on the management of serpiginous choroiditis.

Choroiditis↗

Recent developments in the management of choroidal melanoma.

PURPOSE OF REVIEW: To describe recent advances in the management of choroidal melanoma. RECENT FINDINGS: During the past 10 years, there has been a trend in the management of choroidal melanoma from enucleation to conservative nonenucleation treatments such as thermotherapy, plaque radiotherapy, charged-particle radiotherapy, and local resection. Recently published information regarding conservative management has revealed that plaque radiotherapy combined with thermotherapy offers 97% tumor control, similar to charged-particle radiotherapy. Additionally, patient survival after treatment of medium-size melanoma is similar when comparing plaque radiotherapy versus enucleation. Risk factors identifying small choroidal melanoma include patient symptoms, tumor thickness more than 2 mm, presence of associated subretinal fluid, presence of orange pigment on the tumor surface, and location of the tumor margin at the optic disc. These factors assist in early detection of choroidal melanoma. SUMMARY: The management of choroidal melanoma continues to improve with earlier detection of tumors and better treatment methods. Local tumor control is excellent with radiotherapeutic methods. Future studies should investigate control of subclinical metastasis.

Choroid Neoplasms↗

CNS grafts of rat choroid plexus protect against cerebral ischemia in adult rats.

The present study examined the neuroprotective effects of choroid plexus isolated from adult rats and encapsulated within alginate microcapsules. In vitro, conditioned media from cultured choroid plexus produced a marked, dose-dependent protection of embryonic cortical neurons against serum deprivation-induced cell death. In vivo studies demonstrated that a one-hour middle cerebral artery occlusion in adult Wistar rats produced profound motor and neurological impairments 1-3 days after stroke. In contrast, stroke animals transplanted with encapsulated choroid plexus cells displayed a significant reduction in both motor and neurological abnormalities. Histological analysis 3 days post-transplantation revealed that choroid plexus transplants significantly decreased the volume of striatal infarction. This is the first report demonstrating the therapeutic potential of transplanted choroid plexus for stroke.

Analysis of Variance↗

Combined positron emission tomography/computed tomography for evaluation of presumed choroidal metastases.

BACKGROUND: Choroidal metastases are the most common intraocular malignancy and are the first sign of systemic malignancy in approximately one-third of patients. Of patients with no previous diagnosis of cancer, oncological evaluation fails to find the primary lesion in approximately 50% of cases. Newer imaging modalities such as combined positron emission tomography/computed tomography (PET/CT) may improve the yield of the systemic work-up. METHODS: Consecutive patients presenting with presumed choroidal metastases were evaluated with whole body combined PET/CT scanning. RESULTS: Four patients presenting to a tertiary referral hospital with choroidal metastases as the first sign of systemic malignancy were evaluated. In all four cases, PET/CT demonstrated the ocular lesion, and the primary malignancy which was confirmed by tissue biopsy. False-negative results were seen in two cases of cerebral metastases. PET/CT demonstrated lesions not visible on CT or magnetic resonance imaging in two cases. CONCLUSIONS: Combined PET/CT is a useful addition to the work-up of patients with choroidal metastases. It provides the opportunity to detect lesions not visible with other imaging modalities and the ability to image patients with contraindications to magnetic resonance imaging. It is essential to correlate PET images with clinical information and the results of other imaging modalities and tissue biopsy remains the gold standard in the diagnosis of malignancy. False positives and negatives can occur with PET/CT, and further research is needed before this promising technology becomes a routine part of the evaluation of patients with choroidal metastases.

Adult↗

Transport of prostaglandins and other eicosanoids by the choroid plexus: its characterization and physiological significance.

Choroid plexi from the lateral ventricles of rabbits, cats, and dogfish (Mustelus canis) were used to characterize the prostaglandin (PG) uptake process and to establish its kinetic parameters and substrate specificity. The apparent Kt for PGF2 alpha transport by the rabbit choroid plexus was 20 microM; the Jmax was 27 nmol g-1 min-1. The Ki of inhibition of PGF2 alpha transport by PGE2 was 20 microM; the Jmax of PGF2 alpha transport was unaltered by PGE2. A concentration of p-aminohippuric acid of up to 1 mM did not appreciably affect the Kt or the Jmax of PGF2 alpha transport. The rate of PGF2 alpha accumulation by rabbit choroid plexus was reduced by incubation at 4 degrees C, under anaerobic conditions, in the absence of sodium or in the presence of ouabain, probenecid, or bromcresol green. The choroid plexi of all three species also accumulated thromboxane B2, PGI2, and 6-keto-PGF1 alpha, suggesting that most, if not all, eicosanoids are substrates for this transport system. It is concluded that the choroid plexus transport system satisfies all the criteria of an active, energy-dependent transport system and that this system functions effectively at concentrations of eicosanoids present in the ventricular system under normal or pathological conditions. Hence, this transport system must make an important contribution to the pharmacokinetics of eicosanoids within the brain.

6-Ketoprostaglandin F1 alpha↗

Agonist-induced phosphoinositide hydrolysis in choroid plexus.

5-Hydroxytryptamine (5-HT, serotonin) stimulates phosphoinositide hydrolysis in choroid plexus by interacting with the 5-HTlc site. In the present study, the effects of 5-HT were compared with those of other agonists. 5-HT stimulates a rapid release of all three inositol sugars in a mianserin-sensitive manner. Inositol bisphosphate and inositol trisphosphate levels increase about twofold within 2.5 min, whereas inositol monophosphate levels are not appreciably elevated until 5 min. In contrast, glutamate, carbachol, histamine, substance P, and vasopressin, agents that increase phosphoinositide hydrolysis in other tissues, do not stimulate this response in choroid plexus. High concentrations of norepinephrine increase inositol phosphate release in choroid plexus, but this effect is apparently mediated by activation of the 5-HTlc site. The depolarizing agents KCl and veratrine also fail to stimulate phosphoinositide hydrolysis in choroid plexus. These results, combined with the finding that the phosphoinositide response to 5-HT is insensitive to tetrodotoxin, suggest that the effects of 5-HT are not secondary to neurotransmitter release. Furthermore, an indirect effect mediated via arachidonic acid metabolism is unlikely, since inhibitors of cyclooxygenase and lipoxygenase do not reduce the 5-HT response. We conclude, therefore, that phosphoinositide hydrolysis is the transducing mechanism of the 5-HT 5-HTlc receptor and that the choroid plexus will serve as a useful model system for studies of this receptor.

Animals↗

Nucleoside transporter of cerebral microvessels and choroid plexus.

The nucleoside transporter of cerebral microvessels and choroid plexus was identified and characterized using [3H]nitrobenzylthioinosine (NBMPR) as a specific probe. [3H]NBMPR bound reversibly and with high affinity to a single specific site in particulate fractions of cerebral microvessels, choroid plexus, and cerebral cortex of the rat and the pig. The dissociation constants (KD 0.1-0.7 nM) were similar in the various tissue preparations from each species, but the maximal binding capacities (Bmax) were about fivefold higher in cerebral microvessels and choroid plexus than in the cerebral cortex. Nitrobenzylthioguanosine and dipyridamole were the most potent competitors for [3H]NBMPR binding. Several naturally occurring nucleosides displaced specific [3H]NBMPR binding to cerebral microvessels in vitro, in a rank order that correlated well with their ability to cross the blood-brain barrier in vivo. Adenosine analogues and theophylline were less effective in displacing [3H]NBMPR binding than in displacing adenosine receptor ligands. Photoactivation of cerebral microvessels and choroid plexus bound with [3H]NBMPR followed by solubilization and polyacrylamide gel electrophoresis labeled a protein(s) with a molecular weight of approximately 60,000. These results indicate that cerebral microvessels and choroid plexus have a much higher density of the nucleoside transporter moiety than the cerebral cortex and that this nucleoside transporter has pharmacological properties and a molecular weight similar to those of erythrocytes and other mammalian tissues.

Adenosine↗

High activities of glutamine transaminase K (dichlorovinylcysteine beta-lyase) and omega-amidase in the choroid plexus of rat brain.

Certain halogenated hydrocarbons, e.g., dichloroacetylene, are nephrotoxic to experimental animals and neurotoxic to humans; cysteine-S-conjugate beta-lyases may play a role in the nephrotoxicity. We now show that with dichlorovinylcysteine as substrate the only detectable cysteine-S-conjugate beta-lyase in rat brain homogenates is identical to glutamine transaminase K. The predominant (mitochondrial) form of glutamine transaminase K in rat brain was shown to be immunologically distinct from the predominant (cytosolic) form of the enzyme in rat kidney. Glutamine transaminase K and omega-amidase (constituents of the glutaminase II pathway) activities were shown to be widespread throughout the rat brain. However, the highest specific activities of these enzymes were found in the choroid plexus. The high activity of glutamine transaminase K in choroid plexus was also demonstrated by means of an immunohistochemical staining procedure. Glutamine transaminase K has a broad specificity toward amino acid and alpha-keto acid substrates. The omega-amidase also has a broad specificity; presumably, however, the natural substrates are alpha-ketoglutaramate and alpha-ketosuccinamate, the alpha-keto acid analogues of glutamine and asparagine, respectively. The high activities of both glutamine transaminase K and omega-amidase in the choroid plexus suggest that the two enzymes are linked metabolically and perhaps are coordinately expressed in that organ. The data suggest that the natural substrate of glutamine transaminase K in rat brain is indeed glutamine and that the metabolism of glutamine through the glutaminase II pathway (i.e., L-glutamine and alpha-keto acid-->alpha-ketoglutarate and L-amino acid + ammonia) is an important function of the choroid plexus. Moreover, the present findings also suggest that any explanation of the neurotoxicity of halogenated xenobiotics must take into account the role of glutamine transminase K and its presence in the choroid plexus.

Amidohydrolases↗

Influence of thyroid hormones on transport function and Na+-K+-ATPase activity in the rat choroid plexus.

The sympathomimetic stimulation of choroid plexus transport and secretion in rat seems to be mediated by beta-adrenergic receptors. In the present report the effect of induced changes in thyroid function on transport mechanisms in the rat choroid plexus was studied. Following induction of hyperthyroidism (treatment with T3 for 10 days) the tissue/medium ratio (T/M) for choline uptake in choroid plexus in vitro decreased significantly by 68%. The Na+-K+-ATPase activity showed a statistically significant increase of about 16%. Also following cervical sympathectomy, T3 caused a reduction of the T/M for choline, to the same level as in the non-sympathectomized animals, while the effect of T3 on the Na+-K+-ATPase activity was changed into a 22% decrease. Hypothyroidism (administration of PTU in the drinking water) had little or no effect on the uptake and accumulation of choline in the choroid plexus. The Na+-K+-ATPase activity was reduced by 40%, in contrast to the stimulating effect of T3. There is, hence, reason to believe that the transport of choline in the choroid plexus is only partly regulated by adrenergic mechanisms acting via Na+-K+-ATPase. The major effect of T3 on the choline uptake may be exerted by a mechanism different from the ATPase activity and not involving adrenergic receptors.

Animals↗

Neutral amino acid uptake by the isolated perfused sheep choroid plexus.

1. The uptake of neutral amino acids from the blood into the cells of the choroid plexus was studied by means of the rapid (less than 40 s) single-circulation paired-tracer dilution technique in the isolated perfused choroid plexus of the sheep. 2. The study provides the first direct evidence for the carrier-mediated entry of neutral amino acids from blood into the cells of the choroid plexus. 3. In the terms of Christensen's classification the presence of L-amino acid carrier systems for large neutral amino acids with bulky side chains has been demonstrated. 4. No measurable uptake of [14C]methyl amino isobutyric acid ([14C]MeAIB) during a single passage through the choroid plexus circulation was demonstrated which indicates the probable absence of a significant 'A' transport system. 5. The uptake of small neutral amino acids such as glycine and L-alanine was shown to be carrier-mediated. Results suggest that these amino acids are mainly transported by the glycine and ASC carrier systems, respectively. 6. The results suggest that there is a similarity between the transport systems for neutral amino acids on the blood side of both the blood-brain barrier and blood-cerebrospinal fluid barrier, the exception being for the presence of a glycine carrier on the blood side of the choroid plexus.

Amino Acids↗

Twelve hour reproducibility of choroidal blood flow parameters in healthy subjects.

AIMS/BACKGROUND: To investigate the reproducibility and potential diurnal variation of choroidal blood flow parameters in healthy subjects over a period of 12 hours. METHODS: The choroidal blood flow parameters of 16 healthy non-smoking subjects were measured at five time points during the day (8:00, 11:00, 14:00, 17:00, and 20:00). Outcome parameters were pulsatile ocular blood flow as assessed by pneumotonometry, fundus pulsation amplitude as assessed by laser interferometry, blood velocities in the opthalmic and posterior ciliary arteries as assessed by colour Doppler imaging, and choroidal blood flow, volume, and velocity as assessed by fundus camera based laser Doppler flowmetry. The coefficient of variation and the maximum change from baseline in an individual were calculated for each outcome parameter. RESULTS: None of the techniques used found a diurnal variation in choroidal blood flow. Coefficients of variation were within 2.9% and 13.6% for all outcome parameters. The maximum change from baseline in an individual was much higher, ranging from 11.2% to 58.8%. CONCLUSIONS: These data indicate that in healthy subjects the selected techniques provide adequate reproducibility to be used in clinical studies. Variability may, however, be considerably higher in older subjects or subjects with ocular disease. The higher individual differences in flow parameter readings limit the use of the techniques in clinical practice. To overcome problems with measurement validity, a clinical trial should include as many choroidal blood flow outcome parameters as possible to check for consistency.

Adult↗

Effect of hypertension on foveolar choroidal haemodynamics.

AIMS: To assess the effect of systemic hypertension on the choroidal circulation in subjects with a normal eye examination. METHODS: Laser Doppler flowmetry was used to determine relative choroidal blood velocity (Ch(vel)), volume (Ch(vol)), and flow (Ch(flow)) in the centre of the fovea. Measurements were obtained in 15 eyes of 15 subjects with systemic hypertension. The mean average duration of hypertension was 11 (SD 6) years. Findings obtained in these patients were compared with those of 15 eyes of 15 age matched healthy controls. All subjects had normal eye examinations. RESULTS: No significant differences in Ch(vel), Ch(vol), and Ch(flow), were found between the subjects with and without systemic hypertension despite a 17% higher perfusion pressure (PP) in patients with hypertension. No significant correlation was found between mean arterial pressure or PP and the choroidal circulatory parameters. It was estimated that there is a 90% power to detect a 35% difference in Ch(flow) between the two groups. CONCLUSION: Systemic hypertension does not seem to have a large effect on the choroidal circulation in hypertensive patients that are controlled by antihypertensive therapy. Further studies are needed to elucidate whether systemic hypertension has an effect on the choroidal circulation in patients with ocular diseases.

Adult↗

Photodynamic therapy of circumscribed choroidal haemangioma.

AIM: To evaluate efficacy of verteporfin ocular photodynamic therapy (PDT) in treatment of 10 patients with a symptomatic circumscribed choroidal haemangioma. DESIGN: Prospective non-randomised, interventional case series and critical review of previously published studies. METHODS: 10 consecutive patients (seven primary, two failed transpupillary thermotherapy (TTT), and one failed external beam radiotherapy) with symptomatic circumscribed choroidal haemangioma were treated using verteporfin 6 mg/m2 given as an intravenous infusion over 10 minutes. Diode laser (690 nm) with an intensity of 600 mW/cm2 for 83 seconds (50 J/cm2) was applied 5 minutes after completion of infusion. Single or multiple partially overlapping spots were applied based on the tumour basal dimensions. Periodic follow up with ophthalmoscopy, ultrasonography, and angiographic studies was performed. RESULTS: All 10 patients showed evidence of regression with flattening of tumour, resolution of subretinal fluid, and reduction of choroidal vasculature on angiograms. The visual acuity either improved or remained stable in eight (80%) patients. Visual loss due to delayed choroidal atrophy was seen in two patients. CONCLUSIONS: Although verteporfin PDT is an effective treatment for management of symptomatic circumscribed choroidal haemangioma, delayed treatment related effects can lead to visual loss.

Adult↗

PET/CT imaging: detection of choroidal melanoma.

AIM: To determine the size of untreated choroidal melanomas resolved by whole body positron emission tomography fused with computed tomography (PET/CT). METHODS: 50 consecutive patients with untreated choroidal melanomas underwent whole body PET/CT. A functionally fused helical CT scan and 18-fluoro-2-deoxyglucose (FDG) PET scans were employed. The tumours were identified (both quantitatively and qualitatively) and compared with clinical measurements derived from ophthalmoscopic, angiographic, and ultrasonographic imaging. Standardised uptake values (SUV) of more than 2.5 were considered positive. RESULTS: Among the 50 patients with choroidal melanoma, PET/CT scan SUVs of more than 2.5 were noted in 14 (28%) tumours. No AJCC T1 class tumours, 33.3% of T2 melanomas, and 75% of T3 melanomas were physiologically identifiable on PET/CT. With respect to COMS group classifications, no small choroidal tumours, 33% of medium, and 75% of large melanomas were physiologically identifiable. The sole ring melanoma was identifiable on PET/CT imaging. The smallest tumour physiologically identifiable by PET/CT had basal dimensions of 3x5.9 and an apical height of 2.9 mm. CONCLUSION: Though PET/CT was found to be capable of physiologically identifying certain medium (T2) and most large sized (T3) choroidal melanomas, physiological imaging was not completely dependent upon tumour size. Functionally fused PET/CT localised the tumours within the eye and assessed their physiological activity.

Adult↗

Whole body PET/CT for initial staging of choroidal melanoma.

AIM: To investigate the value of whole body positron emission tomography/computed tomography (PET/CT) in screening for metastatic choroidal melanoma in patients initially diagnosed with choroidal melanoma. METHODS: 52 patients with choroidal melanoma underwent whole body PET/CT as part of their metastatic investigation. PET/CT scans were used as a screening tool at the time of their initial diagnosis. A physical examination, liver function tests, and a baseline chest x ray were also obtained. PET/CT images (utilising intravenous18-fluoro-2-deoxyglucose (FDG)) were studied for the presence of metastatic melanoma. The standards for reference were further imaging and/or subsequent biopsies. RESULTS: Two of 52 (3.8%) patients were found to have metastatic melanoma before treatment. The most common sites for metastases were the liver (100%), bone (50%), and lymph nodes (50%). Brain involvement was also present in one patient. One patient (50%) had involvement of multiple sites. Haematological liver enzyme assays were normal in both patients. PET/CT showed false positive results in three patients (5.7%) when further evaluated by histopathology and/or additional imaging. In seven patients (13.4%) PET/CT imaging detected benign lesions in the bone, lung, lymph nodes, colon, and rectum. CONCLUSION: PET/CT imaging can be used as a screening tool for the detection and localisation of metastatic choroidal melanoma. Liver enzyme assays did not identify liver metastases, while PET/CT revealed both hepatic and extrahepatic metastatic melanoma. PET/CT imaging may improve upon the conventional methods of screening for detection of metastatic disease in patients initially diagnosed with choroidal melanoma.

Adult↗

Decreased prevalence of asymptomatic choroidal metastasis in disseminated breast and lung cancer: argument against screening.

AIM: To determine the frequency of visually asymptomatic choroidal metastases in patients with disseminated breast and lung carcinomas in order to establish optimal patient management policies. METHODS: All patients with confirmed metastatic disease treated in our institution between January 2002 and December 2003 were invited to undergo a funduscopic examination and a B-scan ultrasound evaluation. RESULTS: Of the 169 study participants, 77 had breast cancer (64 with metastases in one organ and 13 with multiple-organ involvement) and 92 had lung cancer (85 with metastases in one organ and 7 with multiple-organ involvement). No patient with metastatic breast cancer and two patients with metastatic lung disease (each with multiple-organ involvement) were found to have choroidal metastases. The choroidal metastases were detected by both the funduscopic and ultrasound examinations. CONCLUSIONS: The 2.17% incidence of choroidal metastasis in disseminated lung cancer and the 0% incidence in disseminated breast cancer speaks against the practicality of screening for early detection of choroidal metastasis among these patients, even though it would lead to early implementation of appropriate, often vision saving, therapeutic management. Its low incidence probably testifies to progress achieved by enhanced systemic oncological treatment policies that have been introduced into routine patient management over the past few years.

Adult↗