Release rate is a key variable affecting the therapeutic effectiveness of liposomal fasudil for the treatment of cerebral ischemia/reperfusion injury.

Liposomal fasudil as a treatment for cerebral ischemia/reperfusion (I/R) injury has been demonstrated to be effective in animal models due to the high accumulation of liposomes in damaged brain tissue. However, it is still unclear what effect drug release rate has on the treatment of I/R injury, where pathology progresses dramatically in a short time. In the present study, we assessed four formulations of liposomal fasudil. The results of an in vitro drug release assay showed that the release properties of fasudil were changed by varying the lipid composition and internal phase of the liposomes. Based on these results, differences in the transition of fasudil plasma concentration were monitored after the administration of each type of liposomal fasudil in normal rats. A pharmacokinetic study showed that higher levels of drug retention in liposomal fasudil resulted in higher fasudil plasma concentration. Finally, treatment of I/R injury model rats with liposomal fasudil revealed that a mid-level release rate of fasudil from liposomes resulted in the greatest therapeutic effect among the formulations. In conclusion, these results demonstrate that an optimized drug release rate from liposomes enhances the therapeutic effect of fasudil for the treatment of cerebral I/R injury.

Intravitreal Aflibercept Injection for Macular Edema Resulting from Central Retinal Vein Occlusion: One-Year Results of the Phase 3 GALILEO Study.

PURPOSE: To evaluate the efficacy and safety of intravitreal aflibercept injections for treatment of macular edema secondary to central retinal vein occlusion (CRVO). DESIGN: A randomized, multicenter, double-masked phase 3 study. PARTICIPANTS: A total of 177 treatment-naive patients with macular edema secondary to CRVO were randomized in a 3:2 ratio. METHODS: Patients received either 2-mg intravitreal aflibercept or sham injections every 4 weeks for 20 weeks. From week 24 to 48, the aflibercept group received aflibercept as needed (pro re nata [PRN]), and the sham group continued receiving sham injections. MAIN OUTCOME MEASURES: The primary efficacy end point was the proportion of patients who gained 15 letters or more in best-corrected visual acuity (BCVA) at week 24. This study reports week 52 results including the proportion of patients who gained 15 letters or more in BCVA and the mean change from baseline BCVA and central retinal thickness. Efficacy end points at week 52 were all exploratory. RESULTS: At week 52, the mean percentage of patients gaining 15 letters or more was 60.2% in the aflibercept group and 32.4% in the sham group (P = 0.0004). Aflibercept patients, compared with sham patients, had a significantly higher mean improvement in BCVA (+16.9 letters vs. +3.8 letters, respectively) and reduction in central retinal thickness (-423.5 µm vs. -219.3 µm, respectively) at week 52 (P < 0.0001 for both). Aflibercept patients received a mean of 2.5 injections (standard deviation, 1.7 injections) during PRN dosing. The most common ocular adverse events in the aflibercept group were related to the injection procedure or the underlying disease, and included macular edema (33.7%), increased intraocular pressure (17.3%), and eye pain (14.4%). CONCLUSIONS: Treatment with intravitreal aflibercept provided significant functional and anatomic benefits after 52 weeks as compared with sham. The improvements achieved after 6 monthly doses at week 24 largely were maintained until week 52 with as-needed dosing. Intravitreal aflibercept generally was well tolerated.

Clinical significance of serum angiopoietin-1 in malignant peritoneal mesothelioma.

We have previously reported that angiopoietin-1 was correlated with pulmonary fibrosis. Here, we investigated the serum levels of angiopoietin-1 in patients with malignant peritoneal mesothelioma, which originate from mesenchymal cells similar to lung fibroblasts. We showed that patients with peritoneal mesothelioma had significantly higher serum levels of angiopoietin-1 in comparison with a population with a history of asbestos exposure without peritoneal mesothelioma, and the Kaplan-Meier method revealed a significant correlation between serum angiopoietin-1 levels and survival. This is the first report about the relationship between angiopoietin-1 and peritoneal mesothelioma.

Mesothelioma cell proliferation through autocrine activation of PDGF-ßß receptor.

BACKGROUND/AIMS: Growth factors play a critical role in proliferation for a variety of cancer cells. The present study was conducted to understand the signaling cascades underlying PDGF-D/PDGF-ßß receptor-mediated proliferation of mesothelioma cells. METHODS: Cell growth and cell cycle were analyzed in human non-malignant Met5A cells and malignant mesothelioma cells such as MSTO-211H, NCI-H28, NCI-H2052, and NCI-H2452 cells. RESULTS: Growth of all the cells used here was not affected by PDGF-D, regardless of concentrations (1-30 ng/ml) or treatment time (48-72 h). Spontaneous growth of those cells was significantly inhibited by knocking-down PDGFD or PDGF-ßß receptor, without affecting cell cycling. The cell growth was significantly inhibited by the Akt inhibitor MK2206 and the ROCK inhibitor Y27632 for all the cell types, by the PDK1 inhibitor BX912 for NCI-H28 cells alone, and by the Rac1 inhibitor NSC23766 for NCI-H2052 cells alone, while the PI3 kinase inhibitor wortmannin had no effect. The cell growth, alternatively, was significantly attenuated by MAP kinase kinase inhibitor PD98059 or the ERK1/2 inhibitor FR180204 for all the cell types. CONCLUSION: The results of the present study show that PDGF-D promotes mesothelioma cell proliferation by targeting ROCK or MAP kinase through autocrine activation of PDGF-ßß receptor.

[Assessment of visual acuity in patients with age-related macular degeneration 1 year after photodynamic therapy].

PURPOSE: To evaluate visual acuity in patients who underwent photodynamic therapy (PDT) for exudative age-related macular degeneration (AMD). METHODS: 138 eyes undergoing PDT were studied retrospectively. 101 eyes with AMD (AMD group) and 37 eyes with polypoidal choroidal vasculopathy (PCV group) were evaluated. RESULTS: In the AMD group, one year after PDT, the log MAR visual acuity improved by 2 lines or more in 23 eyes (22.8%), and decreased by 2 lines or more in 27 eyes (26.7%). In the PCV group, one year after PDT the log MAR visual acuity improved by 2 lines or more in 13 eyes (35.1%), and decreased by 2 lines or more in 8 eyes (21.6%). Out of the 138 eyes, 30 eyes showed improvement in visual acuity by 0.6 or more, one year after PDT (AMD group : 17 eyes; PCV group: 13 eyes) CONCLUSIONS: Patients with PCV who had good primary visual acuity improved their visual acuity by 0.6 or more one year after PDT.

Atypical VZV Retinitis in a Patient with Good Syndrome.

PURPOSE: To describe atypical varicella zoster virus (VZV) retinitis in a patient with Good syndrome. METHODS: A 63-year-old patient with Good syndrome presented with bilateral necrotizing retinitis starting from the posterior pole. He had a history of thymoma status post thymectomy 4 years previously, left-sided sinusitis, and recent pulmonary aspergillosis. Qualitative PCR was performed on aqueous fluid. RESULTS: Immunological investigations revealed reduced levels of CD4+ T cells and immunoglobulins. Qualitative PCR was positive for VZV and negative for cytomegalovirus, herpes simplex virus (HSV)-1, and HSV-2. The patient was treated with oral valacyclovir and three courses of immunoglobulin supplementation. The atypical retinitis showed improvement after therapy. CONCLUSION: Good syndrome should be considered in a patient with opportunistic infections and history of thymoma in the absence of human immunodeficiency virus. Atypical retinitis can occur in patients with Good syndrome and quantitative PCR is important for accurate diagnosis.

[Improvement of optical system for better visibility and intensity of radiation during transpupillary thermotherapy].

PURPOSE: Coagulation is the most serious complication of transpupillary thermotherapy(TTT) for choroidal neovascularization (CNV). To detect coagulation quickly, it is important to have good visibility of the fundus during exposure. With the existing laser optical system using a half mirror, the fundus image is dim, and coagulation is difficult to detect. We have improved the laser optical system for TTT. The purpose of this study was to improve the optical system and to estimate the usefulness of our new system compared with the current optical system. METHOD: We exchanged the half mirror used in the existing TTT optical system to a split mirror. The illumination intensities were measured and the visibility of fundus images were compared. RESULTS: Illumination intensity was increased by means of the split mirror of the slit-lamp for the TTT laser optical system. The fundus image became brighter and clearer. CONCLUSIONS: Improvement of the optical system was useful for safe and effective treatment of cases of CNV with an indication for TTT.

Complement factor H polymorphisms in Japanese population with age-related macular degeneration.

PURPOSE: To study the frequency of five haplotypes previously reported in the complement factor H (CFH) gene for Japanese patients with age-related macular degeneration (AMD). METHODS: Genomic DNA was isolated from peripheral blood samples taken from 96 Japanese AMD patients and 89 age-matched controls. All patients were diagnosed as having exudative (wet-type) AMD. The amplified polymerase chain reaction (PCR) products of CFH exons 2, 9, and 13, and intron 6 were analyzed by temperature gradient capillary electrophoresis (TGCE) and by direct sequencing. The haplotypes were identified, and their frequencies were calculated and compared with reported results. RESULTS: Five haplotypes were identified in the Japanese population including four already reported in the American population. The frequencies of these haplotypes were significantly different between Japanese and American in both control and case groups. The haplotype containing Y402H, which was previously reported to be associated with AMD, was only 4% in the control and case population, with a p value of 0.802. However, two other haplotypes were found as risk factors, which gave an increased likelihood of AMD of 1.9 and 2.5 fold (95% CI 1.12-3.69 and 1.42-6.38). One protective haplotype that decreased the likelihood of AMD by 1.6 fold (95% CI 0.26-0.67) was identified. CONCLUSIONS: The frequencies for five haplotypes previously identified were analyzed in a Japanese population with AMD. Four previously found haplotypes were identified and one additional haplotype was found. The frequencies of each haplotype were significantly different from that in found Americans affected with AMD. Two of the haplotypes were identified as risk factors and one was considered protective.

Is transpupillary thermotherapy applicable to the treatment of age-related macular degeneration with pigment epithelial detachment?

The cause of pigment epithelial tears at the edge of a pigment epithelial detachment (PED) following transpupillary thermotherapy (TTT) in eyes with a PED associated with age-related macular degeneration has not been conclusively determined. We have treated two eyes that had a PED with TTT. A pigment epithelial tear developed in one eye but not in the other. Our findings suggest that pigment epithelial tears are probably related to the shape of the PED, and TTT should not be applied to a balloon-shaped PED.

Isolated metastases of hepatocellular carcinoma in the left atrium, unresponsive to treatment with sorafenib.

Isolated metastases in the left atrium that are discontinuous with an intrahepatic hepatocellular carcinoma (HCC) are extremely rare. This is the case report of a 46-year-old male patient with pulmonary metastases from HCC, who presented with a tumor in the left lung, extending to the left atrium through the left pulmonary vein. Two weeks after the initiation of treatment with sorafenib, the tumor metastasized to the left parietal cerebral lobe, with an intracranial hemorrhage. Although the patient underwent gamma knife radiosurgery for the metastatic brain tumor, his condition gradually deteriorated and he succumbed to multiple organ failure 4 months later. Given the severe complications that have been reported in patients with this type of metastasis, immediate multidisciplinary treatment, including surgical resection, should be considered.

Efficacy and safety of intravitreal aflibercept injection in wet age-related macular degeneration: outcomes in the Japanese subgroup of the VIEW 2 study.

BACKGROUND/AIMS: To evaluate efficacy and safety of intravitreal aflibercept (IVT-AFL) in Japanese patients with wet age-related macular degeneration (wAMD) from the VIEW 2 trial. METHODS: In this double-masked study, patients were randomised to: 0.5 mg IVT-AFL every 4 weeks (0.5q4); 2 mg IVT-AFL every 4 weeks (2q4); 2 mg IVT-AFL every 8 weeks (2q8) after 3 monthly injections; or 0.5 mg ranibizumab every 4 weeks (Rq4). Main efficacy outcomes included vision maintenance and best-corrected visual acuity (BCVA) at week 52. RESULTS: At week 52, all Japanese patients in the IVT-AFL groups (n=70) maintained vision, compared with 96% of Japanese patients (n=23/24) treated with ranibizumab. Japanese patients in all treatment groups showed improvement in BCVA after treatment. The Rq4, 2q4 and 2q8 groups experienced similar gains in BCVA from baseline. The 0.5q4 group had higher gains due to an unexpected drop in BCVA between screening and baseline. Central retinal thickness and mean area of choroidal neovascularisation decreased in all treatment groups with similar magnitude. Ocular treatment-emergent adverse events were balanced across treatment groups. CONCLUSIONS: IVT-AFL was effective and well tolerated in Japanese patients. Outcomes in this population were consistent with those in the overall VIEW 2 population. TRIAL REGISTRATION NUMBER: NCT00637377.

Liposomes and nanotechnology in drug development: focus on ocular targets.

Poor drug delivery to lesions in patients' eyes is a major obstacle to the treatment of ocular diseases. The accessibility of these areas to drugs is highly restricted by the presence of barriers, including the corneal barrier, aqueous barrier, and the inner and outer blood-retinal barriers. In particular, the posterior segment is difficult to reach for drugs because of its structural peculiarities. This review discusses various barriers to drug delivery and provides comprehensive information for designing nanoparticle-mediated drug delivery systems for the treatment of ocular diseases. Nanoparticles can be designed to improve penetration, controlled release, and drug targeting. As highlighted in this review, the therapeutic efficacy of drugs in ocular diseases has been reported to be enhanced by the use of nanoparticles such as liposomes, micro/nanospheres, microemulsions, and dendrimers. Our recent data show that intravitreal injection of targeted liposomes encapsulating an angiogenesis inhibitor caused significantly greater suppression of choroidal neovascularization than did the injection of free drug. Recent progress in ocular drug delivery systems research has provided new insights into drug development, and the use of nanoparticles for drug delivery is thus a promising approach for advanced therapy of ocular diseases.

Suppression of choroidal neovascularization by intravitreal injection of liposomal SU5416.

OBJECTIVE: To clarify whether use of angiogenic vessel-homing peptide, Ala-Pro-Arg-Pro-Gly (APRPG)-modified liposomes encapsulating 3-([2,4-dimethylpyrrhol-5-yl] methylidenyl)-indolin-2-one (SU5416), an angiogenesis inhibitor, can inhibit the development of experimental choroidal neovascularization (CNV) in rats. METHODS: Liposomes were prepared using the thin-film hydration method. To set up the rat experimental CNV model, intense fundus laser photocoagulation at 6 spots per eye was performed on pigmented rats. After photocoagulation, the rats were divided into 4 groups (6 rats in each group): an APRPG-liposomal SU5416 treatment group and control groups treated with a balanced salt solution, APRPG liposomes, or soluble SU5416. Each rat received a single intravitreal injection immediately after the injury. One week or 2 weeks after laser injury, the extent of CNV was evaluated by perfusion with high-molecular-weight fluorescein isothiocyanate-dextran. RESULTS: Two weeks after injection, the CNV area was significantly (P < .05) smaller in the APRPG-liposomal SU5416-treated group compared with the CNV area in the balanced salt solution-and APRPG liposome-treated groups. CONCLUSION: Liposomes modified by APRPG and encapsulating SU5416 constitute a potential drug formulation for CNV treatment that would require only a single intravitreal injection. CLINICAL RELEVANCE: This liposomal delivery may enable the sustained release of small molecules and be a new treatment modality for CNV.

Genetic analysis of typical wet-type age-related macular degeneration and polypoidal choroidal vasculopathy in Japanese population.

Age-related macular degeneration (AMD) is a common cause of blindness in the elderly. Caucasian patients are predominantly affected by the dry form of AMD, whereas Japanese patients have predominantly the wet form of AMD and/or polypoidal choroidal vasculopathy (PCV). Although genetic association in the 10q26 (ARMS2/HTRA1) region has been established in many ethnic groups for dry-type AMD, typical wet-type AMD, and PCV, the contribution of the 1q32 (CFH) region seem to differ among these groups. Here we show a single nucleotide polymorphism (SNP) in the ARMS2/HTRA1 locus is associated in the whole genome for Japanese typical wet-type AMD (rs10490924: p = 4.1 x 10(-4), OR = 4.16) and PCV (rs10490924: p = 3.7 x 10(-8), OR = 2.72) followed by CFH (rs800292: p = 7.4 x 10(-5), OR = 2.08; p = 2.6 x 10(-4), OR = 2.00), which differs from previous studies in Caucasian populations. Moreover, a SNP (rs2241394) in complement component C3 gene showed significant association with PCV (p = 2.5 x 10(-3), OR = 3.47). We conclude that dry-type AMD, typical wet-type AMD, and PCV have both common and distinct genetic risks that become apparent when comparing Japanese versus Caucasian populations. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12177-009-9047-1) contains supplementary material, which is available to authorized users.

Measurement of retinal thickness from three-dimensional images obtained from C scan images from the optical coherence tomography ophthalmoscope.

BACKGROUND: One of the unique features of the optical coherence tomography (OCT) ophthalmoscope is that it can record C scan images of the retina. The purpose of this study was to determine the best recording time to measure the retinal thickness with the OCT ophthalmoscope. In addition, the accuracy of the measurements was examined by comparing the values obtained by the OCT ophthalmoscope with those obtained with the Stratus OCT Model 3000 (OCT III) assuming that the OCT III gives an accurate measurement of retinal thickness. METHODS: The topography mode of the OCT ophthalmoscope was used. First, the average retinal thickness recorded with 2-s scans was compared with that recorded with 4-s scans for a recording area of 15 degreesx15 degrees. Next, the average retinal thickness recorded by the OCT ophthalmoscope was compared with that obtained by the Fast Macular Thickness program of the OCT III in patients with macular oedema. RESULTS: The mean retinal thickness of the central area was 208.1 microm for both 2 and 4 s recording times. The average retinal thickness obtained by the OCT ophthalmoscope was highly correlated and not significantly different from the values obtained by the OCT III. The largest differences obtained by the two instruments were seen in the parts of the retina with accumulation of hard exudates. CONCLUSIONS: Accurate measurements of retinal thickness can be obtained with the OCT ophthalmoscope by 2-s scans, and thus, the OCT ophthalmoscope can be a valuable instrument for clinical assessments of retinal thickness.

Is transpupillary thermotherapy applicable to the treatment of age-related macular degeneration with pigment epithelial detachment?

The cause of pigment epithelial tears at the edge of a pigment epithelial detachment (PED) following transpupillary thermotherapy in eyes with associated age-related macular degeneration is unclear. We have treated 2 eyes which had a PED with TTT. Our findings suggest pigment epithelial tears are probably related to the shape of the PED and TTT should not applied to a balloon-shaped PED.