Chieko Shima

Levels of vascular endothelial growth factor and pigment epithelium-derived factor in eyes before and after intravitreal injection of bevacizumab.

PurposeBevacizumab is a human monoclonal IgG1 antibody that blocks the action of vascular endothelial growth factor (VEGF). The purpose of this study was to determine the level of VEGF and pigment epithelium-derived factor (PEDF) in eyes with proliferative diabetic retinopathy (PDR) before and after an intravitreal injection of bevacizumab.MethodsEleven eyes of ten patients were studied. Patients were included if they had neovascular glaucoma, rubeosis of the iris with PDR, or aggressive PDR. Samples of aqueous humor were collected just before the injection of bevacizumab and the vitrectomy. The concentrations of VEGF and PEDF in the aqueous humor were measured by enzyme-linked immunosorbent assay, and the effects of bevacizumab on PDR were evaluated.ResultsThe free VEGF concentration before the injection was 676.5 ± 186.7 pg/ml (mean ± SEM, n = 11). Seven days later, it was significantly reduced to 7.1 ± 7.1 pg/ml (P < 0.005, n = 9). The PEDF concentration before the injection was 2.32 ± 0.49 μg/ml (n = 11), and 7 days later, it was 3.23 ± 0.76 μg/ml (P = 0.33). During the vitrectomy, patients had less intraoperative bleeding when the neovascular tissues were cut.ConclusionsAn intravitreal injection of bevacizumab significantly decreased the free VEGF in the aqueous humor by 7 days, indicating that the clinical effects of bevacizumab appear rapidly. However, intravitreal bevacizumab did not affect the level of intraocular PEDF.

Intraocular Pressure Elevation after Injection of Triamcinolone Acetonide : A Multicenter Retrospective Case-Control Study

PURPOSE To determine the risk factors for intraocular pressure (IOP) elevation after the injection of triamcinolone acetonide (TA). DESIGN Retrospective interventional case-control study. METHODS SETTING Multicenter. PATIENT POPULATION Four hundred and twenty-seven patients. OBSERVATION PROCEDURES Intraocular pressure levels after TA treatment by the sub-Tenon capsule injection (STI; 12 mg, 20 mg, or 40 mg), intravitreal injection (IVI; 4 mg or 8 mg), or the combination of STI (20 mg) and IVI (4 mg), and IOP levels after two TA treatments. MAIN OUTCOME MEASURE Risk factors for IOP levels of 24 mm Hg or higher. RESULTS Younger age (hazards ratio [HR], 0.96/year; P < .0001), IVI (HR, 1.89/year; P < .0001), and higher baseline IOP (HR, 1.15/mm Hg; P = .003) were identified as risk factors. Dose dependency was shown in STI-treated eyes (HR, 1.07/mg; P = .0006), as well as after IVI (HR, 1.64/mg; P = .013). The combination of STI and IVI was a significant risk factor (HR, 2.27; P = .003) compared with STI alone. In eyes receiving two TA treatments, IVI (HR, 2.60; P = .010), higher IOP elevation after the first injection (HR, 1.18/mm Hg; P = .011), and increased dosage of STI (HR, 1.07/mm Hg; P = .033) were risk factors. CONCLUSIONS Younger age, higher baseline IOP, IVI, and increased TA dosage were associated with TA-induced IOP elevation. IOP elevation after repeated TA injection was frequently associated with eyes treated with IVI, high IOP elevation after the first injection, and high doses of STI.

Neuroprotective Effects of Granulocyte Colony-Stimulating Factor on Ischemia-Reperfusion Injury of the Retina

Purpose: It has been reported that granulocyte colony-stimulating factor (G-CSF) provides neuroprotection in models in which neuronal cell death is induced. This research was designed to investigate the effects of G-CSF on neurodegeneration of the inner retinal layer in a rat model of ischemic reperfusion (I/R) injury. Materials and Methods: Retinal ischemia was induced by increasing the intraocular pressure to 110 mm Hg for 45 min in the left eyes of the rats. A sham operation was carried out on the right eyes. G-CSF (100 µg/kg/day in 0.3 ml saline) or the same volume of saline was intraperitoneally injected just before the operation and continued for 4 consecutive days (a total of 5 consecutive days). Morphological examinations, including the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, were performed 7 days after I/R induction. The expression of phosphorylated AKT in the retina was examined by Western blot analysis and immunohistochemistry. Results: Cell loss in the ganglion cell layer was more significantly reduced in the I/R-induced eyes of the G-CSF-injected rats than in the I/R-induced eyes of the saline-injected rats (20.3 vs. 6.6%). The inner retinal thickness ratios, such as the inner plexiform layer to the inner limiting membrane/outer nuclear layer and the inner nuclear layer/outer nuclear layer, were significantly better preserved in the I/R-induced eyes of the G-CSF-injected rats than in the I/R-induced eyes of the saline-injected rats. TUNEL assays showed fewer apoptotic cells in the retinal sections of the I/R-induced eyes of the G-CSF-injected rats. The phosphorylation of AKT (p-AKT/AKT) was upregulated in the retinas of the I/R-induced eyes of the G-CSF-injected rats. Conclusion: Our results demonstrated that systemic injection of G-CSF can protect retinal ganglion cells and inner retinal layers from I/R injury. The effects could be associated with the activation of AKT.

Combination of Intra-Bone Marrow–Bone Marrow Transplantation and Subcutaneous Donor Splenocyte Injection Diminishes Risk of Graft-Versus-Host Disease and Enhances Survival Rate

The combination of allogeneic bone marrow transplantation (allo-BMT) and donor lymphocyte infusion (DLI) is a useful method for establishing donor chimerism and preventing a relapse of leukemia/lymphoma. However, there is a risk of inducing uncontrollable fatal graft-versus-host disease (GVHD). In fact, allo-BMT plus intravenous (IV)-DLI using donor splenocytes induces fatal GVHD in recipient mice. In this study, we examined the effects of the combination of intra-bone marrow (IBM)-BMT and the subcutaneous injection of donor splenocytes (SC-DLI) on the allo-BMT system. Recipient BALB/c mice were conditioned by sublethal irradiation (5 Gy), followed by IBM-BMT plus IV-DLI or SC-DLI in C57BL/6 mice. The IV-DLI group showed better engraftment of donor hemopoietic cells than the control group (without DLI) but showed fatal GVHD. The SC-DLI group, however, showed good reconstitution and mild GVHD. These results suggest that the combination of SC-DLI and IBM-BMT promotes the reconstitution of hemopoiesis and helps reduce the risk of GVHD.

Caspase inhibitor ZVAD-fmk facilitates engraftment of donor hematopoietic stem cells in intra-bone marrow-bone marrow transplantation.

When bone marrow transplantation (BMT) is carried out, survival of the donor hematopoietic stem cells is crucial to maintain donor hematopoiesis in the recipients. We have shown that intra-bone marrow-bone marrow transplantation (IBM-BMT) can induce the rapid recovery of donor hematopoiesis and allow a reduction in radiation doses as a pretreatment for BMT. If IBM-BMT methodology can be further improved, BMT could be carried out more safely and more easily. In this experiment, we attempted to suppress apoptosis of donor hematopoietic cells using a caspase inhibitor, ZVAD-fmk, upon IBM-BMT in mouse allogeneic IBM-BMT. IBM-BMT with ZVAD-fmk induced superior engraftment of donor hematopoietic cells and greater numbers of day-12 colony-forming units of spleen (CFU-S) than IBM-BMT without ZVAD-fmk upon allogeneic BMT (C57BL/6 into BALB/c mice). ZVAD-fmk slightly suppressed apoptosis of whole BMCs, whereas it significantly suppressed apoptosis of c-kit+/Sca-1+/lineage(-) cells (KSL cells) in vitro. These results suggest that ZVAD-fmk can suppress apoptosis of hematopoietic stem cells and/or immature progenitor cells of the donor bone marrow cells, thereby accelerating the donor hematopoiesis.

Dampening of diurnal intraocular pressure fluctuation by combined trabeculotomy and sinusotomy in eyes with open-angle glaucoma.

Purpose:Large diurnal intraocular pressure (IOP) fluctuation ([INCREMENT]IOP) is believed to be one of the causes of progression in glaucomatous changes. Some fully medicated glaucoma patients whose IOPs are controlled during the regular office hours (10:00 to 16:00 h) still have progression in glaucomatous changes and IOP elevation during off-office hours. The purpose of this study was to determine whether [INCREMENT]IOP is dampened after combined trabeculotomy and sinusotomy (LOT+SIN) in glaucoma patients with low IOPs during the regular office hours. Patients and Methods:Fourteen eyes of 8 open-angle glaucoma patients who had large [INCREMENT]IOP despite low IOPs during the office hours were studied. The IOP was measured every 3 hours for 24 hours before and >3 months after the operation. The IOPs were measured in the sitting position with a Goldmann applanation tonometer. All patients underwent LOT+SIN. Results:All patients had IOP elevations >20 mm Hg between 0:00 and 3:00 hours before the operation, and none had an IOP peak after the operation. The postoperative mean IOP (16.5±1.7 to 13.9±2.0 mm Hg, P=0.00064), the maximum IOP (21.9±2.4 to 16.1±2.5 mm Hg, P=0.0020), and [INCREMENT]IOP (8.9±2.7 to 4.3±1.2 mm Hg, P=0.0032) were significantly lower than the preoperative values. However, the minimum IOP was not reduced significantly (13.0±1.9 to 11.7±1.7 mm Hg). Conclusions:The diurnal [INCREMENT]IOPs are dampened by LOT+SIN in glaucoma patients with controlled IOPs during regular office hours. These results indicate that these surgical procedures can be used for the treatment of open-angle glaucoma patients.

Abnormal distribution of dendritic cells in (NZW × BXSB)F1 mice

(NZW × BXSB)F1 mice (W/BF1 mice) have been reported to develop autoimmune diseases with aging. We have also reported that the number of dendritic cells (DCs) increases in the various organs, and that the B-cell response to LPS or interleukin-4 plus anti-μ increase with aging in W/BF1 mice. In the present experiment, we show that many DCs exist not only in the T-cell area but also in the B-cell area and the sinus in the spleen of aged W/BF1 mice, and that the coculturing of DCs from aged W/BF1 mice and B cells from disease-free young W/BF1 mice produces much more IgG and IgM than normal mice. These results suggest that an abnormal distribution of DCs and the interaction of DCs and B cells induce the hyperproduction of immunoglobulin in aged W/BF1 mice.

Posterior sub-Tenon injection of triamcinolone acetonide as pretreatment for focal laser photocoagulation in diabetic macular edema patients

PurposeTo determine whether a posterior sub-Tenon injection of triamcinolone acetonide (TA) before focal photocoagulation is safe and effective in patients with diabetic macular edema.MethodsSixteen eyes of 11 diabetic patients with unresolved diffuse macular edema were treated with a 20-mg sub-Tenon injection of TA 1 to 2 months before focal photocoagulation. Focal photocoagulation was applied only to microaneurysms, and grid laser photocoagulation was not performed. The main outcome measures used were visual acuity (VA), central macular thickness (CMT) determined by optical coherence tomography (OCT), and the fluorescein angiographic appearance of the retina. Patients were followed for at least 6 months.ResultsOne month after the sub-Tenon injection of TA, the macular edema was resolved with a significant reduction of the CMT on OCT. VA improved slightly. Subsequent focal photocoagulation of the microaneurysms maintained the significant reduction of CMT for up to 6 months. A significant improvement of VA was observed in 37.5% patients at 6 months, and there was no decrease in VA in any of the patients.ConclusionsA 20-mg sub-Tenon TA injection prior to focal laser photocoagulation is a safe and beneficial treatment in patients with diabetic macular edema.

Choroidal melanoma with massive extraocular extensions through sclera.

We report a case of choroidal melanoma with massive extraocular extensions through the sclera. A 64-year-old woman reported blurred vision in her right eye. At the first visit, visual acuity was 10/20 OD. An ophthalmological examination revealed a raised choroidal mass and exudative retinal detachment in the lower retina. A diagnosis of peripheral choroidal melanoma was confirmed by additional test results. Because the tumor size was large and no systemic metastasis was found, we recommended enucleation. However, the patient refused and requested only to be followed without treatment. Seven months later, the tumor showed extraocular extensions through the sclera into subconjunctival space and she finally agreed to undergo enucleation. Histopathologic findings showed that the tumor was a mixed cell malignant melanoma of the choroid. The eye was filled with tumor cells, and the tumor had massive extraocular extensions into the orbit through the sclera and scleral emissarium vessels. The intraocular tumor was markedly necrotic, which indicated rapid growth. Choroidal melanomas can increase quickly in size resulting in extraocular extensions through the sclera.