Effects of several anti-glaucoma medications on the circadian intraocular pressure fluctuations in patients with primary open-angle glaucoma.

PURPOSE: The purpose of the present study was to elucidate the effects of several anti-glaucoma medications on the circadian intraocular pressure (IOP) fluctuations in patients with primary open-angle glaucoma (POAG). PATIENTS AND METHODS: POAG patients (n=61; 61 eyes) with or without glaucoma medications were included. IOP measurement at 14 time points (12, 15, 18, 21, 0, 6, 9, 12, 15, 18, 21, 0, 6, and 9 o'clock) was performed over a period of 48 h. IOP changes occurring in the first 24 h and the subsequent 24 h were evaluated by several therapeutic factors. RESULTS: A nocturnal acrophase pattern was observed in all the eyes with POAG. The shape of the first 24 h IOP curve was similar to that of the following 24 h IOP curves. However, there were fewer overall IOP levels in the second 24 h time period. Circadian IOP fluctuation patterns exhibited in each eye on the 1st and 2nd days were single acrophase patterns: diurnal acrophase (1st day, 54.0%; 2nd day, 60.7%) and nocturnal acrophase (1st day, 36.1%; 2nd day, 31.1%), and no single acrophase patterns: flat (1st day, 6.6%; 2nd day, 4.9%) and double acrophase (1st day, 3.3%; 2nd day, 3.3%). Among the different medication groups, a nocturnal acrophase circadian pattern was observed in the patient groups being treated by combinations of prostaglandin analog (PG) and ß blocker or PG, ß blocker and carbonic anhydrase inhibitor (CAI). However, this was not apparent in patient groups with or without single anti-glaucoma medications or a combination of PG and CAI. CONCLUSIONS: The present study of IOP monitoring patients with POAG over a period of 48 h indicated that their changes in circadian patterns of IOP were affected by types of anti-glaucoma medications.

Black currant anthocyanins normalized abnormal levels of serum concentrations of endothelin-1 in patients with glaucoma.

PURPOSE: Our recent study, which involved a randomized, placebo-controlled, double-masked 24-month trial (Ophthalmologica 2012;228:26-35), revealed that oral administration of black currant anthocyanins (BCACs) slowed down the visual field deterioration and elevation of ocular blood flow of open-angle glaucoma (OAG). To elucidate the underlying mechanisms of these BCAC-induced effects, as possible factors affecting glaucomatous optic neuropathy, changes of serum endothelin-1 (ET-1), nitric oxide (NO), and antioxidative activities were examined in the present study. METHODS: From among patients with OAG who participated in the randomized, placebo-controlled, double-masked trial, serum specimens were obtained from BCAC-treated (n=19) or placebo-treated (n=19) patients at baseline and every 6 months. Healthy volunteers (n=20) with age and gender matching the patients were used as a control. Serum ET-1 concentration, [NO2(-)] and [NO2(-) + NO3(-)] levels, advanced oxidation protein products (AOPP), and antioxidant activities were measured by using commercially available kits. RESULTS: At the trial baseline, serum ET-1 concentrations were significantly lower in patients with OAG (BCACs, 3.18±1.06 pg/mL; placebo, 3.44±0.84 pg/mL) than those in healthy volunteers (4.38±1.03 pg/mL) (one-way analysis of variance and a Tukey's multiple comparison post hoc test, P<0.05). Upon administration of BCACs, serum ET-1 concentrations increased to the levels of those in healthy volunteers during the 24-month period. In contrast, those of placebo-treated patients remained at lower levels (3.82±1.14 pg/mL). While [NO2(-)] and [NO2(-)+NO3(-)] levels, AOPP, and antioxidative activities of patients from both the BCACs and placebo groups showed comparable levels to those of healthy subjects at baseline, no significant changes were observed during the observational period in either the BCAC or placebo groups. CONCLUSIONS: Among the possible beneficial effects of BCACs toward visual field progression in patients with OAG, our present results suggest that BCACs caused normalization of serum ET-1 levels, and this may modulate ET-1-dependent regulation of the ocular blood hemodynamics.

Effects of black currant anthocyanins on intraocular pressure in healthy volunteers and patients with glaucoma.

PURPOSE: The purpose of the present study was to elucidate the effects of oral administration of black currant anthocyanins (BCACs) on intraocular pressure (IOP) in both healthy subjects and patients with glaucoma. METHODS: (1) A placebo-controlled, double-masked, crossover study (n=12) was conducted, during which BCACs (50 mg/day) or placebos were orally administered to 12 healthy subjects once daily for 4 weeks. (2) A total of 21 glaucoma patients (BCACs, n=12; placebo, n=9) treated with a single antiglaucoma medication who had participated in a previous study (a randomized, double-masked, placebo-controlled trial, Ophthalmologica 2012) were selected and analyzed. Systemic blood pressure, pulse rates, IOP, and Humphrey visual-field mean deviation (MD) (program 30-2, SITA standard) were evaluated. RESULTS: (1) A statistically significant decrease in the mean IOP was observed at 2 weeks (P=0.002, paired t-test) and 4 weeks (P=0.039, paired t-test) from the baseline in BCAC-treated healthy subjects. This decrease, however, was not observed in the placebo group. In addition, at 2 weeks after the baseline, changes were also statistically significant between the groups (P=0.027, paired t-test). (2) Intergroup and between-group analyses revealed statistically significant decreases in mean IOP in the glaucoma patients taking BCACs (P=0.027, paired t-test; P=0.024, unpaired t-test) at 24 months after the baseline. In addition, mean changes of MD deterioration were significantly less in BCAC glaucoma patients administered with BCACs at 12 months (P=0.017, Mann-Whitney U test) and 18 months (P=0.050, Mann-Whitney U test) after the baseline. No clinically significant changes were observed in systemic blood pressure or pulse rates in either trial. CONCLUSIONS: Our results suggested that oral administration of BCACs may induce a beneficial decrease in IOP levels in healthy subjects as well as in patients with glaucoma.

The effects of a fixed combination of 0.5% timolol and 1% dorzolamide on optic nerve head blood circulation.

PURPOSE: The purpose of the present study is to evaluate the effects of a fixed combination of 0.5% timolol maleate (TM) and 1% dorzolamide hydrochloride (DZ) (FCTD(1%)) on optic nerve head (ONH) blood circulation. SUBJECTS AND METHODS: A drop of 0.5% TM, 1% DZ, or FCTD(1%) was topically administered to the right eyes of 15 healthy Japanese subjects, and a drop of saline was likewise administrated to their left eyes. Systemic blood pressure, heart rates (HRs), intraocular pressures (IOPs), ocular perfusion pressures, and blood circulation by a laser speckle flowgraphy at the ONH were measured before administration, and again at 1.5 and 6 h afterward. RESULTS: FCTD(1%) caused a significant increase in ONH blood circulation 6 h after the administration, while 0.5% TM and 1% DZ had no significant effect. The IOPs were significantly decreased in 0.5% TM (P<0.01) and FCTD(1%) (P<0.05) when compared with the saline-treated eyes at 1.5 and 6 h after administration, while 1% DZ did not show a statistically significant decrease. There were no significant changes in other systemic parameters except for a significant transient decrease in the HRs observed in 0.5% TM (P<0.01) when compared with the baseline at 1.5 h after the administration. CONCLUSION: Our current results suggest that FCTD(1%) caused a significant increase in ocular blood circulation on the ONH in healthy Japanese subjects, presumably by a synergistic effect of 0.5% TM and 1% DZ, although neither 0.5% TM nor 1% DZ on their own had a significant effect.

Two-year randomized, placebo-controlled study of black currant anthocyanins on visual field in glaucoma.

AIM: To examine the influence of the black currant anthocyanins (BCACs) on the disease progression of open-angle glaucoma (OAG), a randomized, placebo-controlled, double-masked trial was made in 38 patients with OAG treated by antiglaucoma drops. METHODS: BCACs (50 mg/day, n = 19) or their placebos (n = 19) were orally administered once daily for a 24-month period. Systemic blood pressure, pulse rates, intraocular pressure (IOP), ocular blood circulation by laser-speckle flowgraphy, and Humphrey visual field mean deviation (MD) were measured during the 24-month period. RESULTS: As a main outcome measurement, we evaluated the difference between the groups in MD deterioration in the eye with a better MD from the trial's baseline through 24 months. A statistically significant difference was observed between the treatment groups in mean change from baseline in MD 24 months after therapy (p = 0.039, unpaired t test). Upon administration of BCACs, the ocular blood flows during the 24-month observational period increased in comparison with placebo-treated patients. However, no significant changes were observed in systemic and ocular conditions including IOP during the 24-month period. CONCLUSIONS: Our results suggest that oral administration of BCACs may be a safe and promising supplement for patients with OAG in addition to antiglaucoma medication.

Lysosomal phospholipase A2 activity in pig aqueous humor.

PURPOSE: The present study was conducted to determine lysosomal phospholipase A2 (LPLA2) activity in the aqueous humor (AH) and to identify the possible sources of the LPLA2 found in the AH. METHODS: To detect LPLA2 activity in pig AH and ocular tissues, liposomes consisting of 1,2-dioleoylphosphatidylglycerol/N-acetylsphingosine were used as substrates in an activity assay under acidic conditions. The reaction products were separated by thin-layer chromatography. To identify the LPLA2 in pig AH, the AH was analyzed by Western immunoblot analysis with an anti-LPLA2 antibody. Distribution of the LPLA2 in pig ocular tissues was studied by determining its activity in individual tissue extracts. RESULTS: LPLA2 activity was detected in the AH obtained from pig eyes. Consistent with the known properties of LPLA2, the activity was heat-labile and undetectable at neutral pH. The immunoblot of pig AH showed the anti-LPLA2 antibody-reactive protein band. In addition, the specific activity of the enzyme, when normalized to volume, was higher in pig AH than in pig serum. Individual tissue extracts obtained from pig ocular tissues showed different specific activity of LPLA2. In particular, the extract prepared from the trabecular meshwork provided the highest specific activity. CONCLUSIONS: The present findings suggest that the phospholipase A2 activity found in pig AH under acidic conditions is due to LPLA2 and that it originates from ocular tissues surrounding the anterior chamber as well as plasma.

Clinical distinction between nasal optic disc hypoplasia (NOH) and glaucoma with NOH-like temporal visual field defects.

PURPOSE: To report on the clinically important differences between nasal optic hypoplasia (NOH) and glaucoma with NOH-like temporal visual field defect (VFD). METHOD: Five NOH (four bilateral and one unilateral) patients, three unilateral NOH patients with glaucoma, and two glaucoma patients with NOH-like temporal VFD were clinically characterized. Superior segmental optic nerve hypoplasia was also associated with glaucoma in one eye of a bilateral NOH case and the NOH eye of a unilateral NOH patient. Ocular manifestations including refractive errors, size, and appearances of the optic discs, retinal nerve fiber thickness (NFLT) ascertained by optical coherence tomography (OCT), and VFD were examined. RESULTS: Ophthalmic examinations revealing NOH showed high myopia at more than -5.0D, a small disc with nasal double-ring appearance, significantly decreased NFLT by OCT, and retinal nerve fiber layer defect in the corresponding nasal sector. Stationary temporal VFD varied from a slight depression of the peripheral isopters to wide sector defects. In contrast, two glaucoma patients with NOH-like temporal VFD showed several different clinical features, including mild myopia less than -5D, a normal size with glaucomatous disc cupping; a slight decrease in nasal NFLT and progression of temporal and other glaucomatous VFD. CONCLUSION: Careful evaluation of optic disc appearance and measurement of NFLT using OCT may help to distinguish between NOH and glaucoma with NOH-like temporal VFD.

Clinicopathological roles of aberrantly expressed recoverin in malignant tumor cells.

PURPOSE: Cancer-associated retinopathy (CAR) is an ocular manifestation of a paraneoplastic syndrome whereby immunological reactions toward recoverin, a retina-specific calcium binding protein, and other retinal antigens aberrantly expressed in tumor cells are elicited. As a consequence, photoreceptor cell degeneration is induced. To elucidate the pathological role of the aberrantly expressed recoverin, we studied the recoverin expression levels in various malignant tumors and the effects of the expressed recoverin on the sensitivity to anti-cancer drugs. METHODS: Recoverin expression levels were determined by immunohistochemistry with anti-human recoverin monoclonal antibody on multiple tissue arrays obtained from several lung, stomach, colon and other cancers. In the presence of several anti-cancer drugs, including anti-tumor antibiotics, plant alkaloids and anti-metabolites, cytotoxicity assay was performed using recoverin-positive or recoverin-negative A549 cells originating from human lung adenocarcinoma. RESULTS: Immunofluorescence labeling revealed that recoverin immunoreactivities were detected at approximately 10-40% of malignant tumor tissues. Cytotoxic effects by anti-cancer drugs were higher in recoverin-positive A549 cells as compared to recoverin-negative cells. CONCLUSION: The present data may correlate with the previous observation that recoverin-expressing cancer cells induced tumor immunity and a favorable prognosis for primary cancer in CAR patients.

Misregulation of rhodopsin phosphorylation and dephosphorylation found in P23H rat retinal degeneration.

To examine rhodopsin (Rho) functions in P23H rat, kinetics of Rho regeneration and dephosphorylation were investigated by spectrophotometric analysis and immunofluorescence labeling method using specific antibodies toward phosphorylated 334Ser or 338Ser site. Rho dephosphorylation at both sites was extremely delayed in P23H retina as compared to normal ones. Kinetics of Rho regeneration was not altered between normal and P23H rats under dark adaptation. Next, to study the effects of several Ca(2+)channel blockers on this model, retinal function and morphology were evaluated. Among them, nilvadipine showed a significant protective effect against P23H retinal degeneration. Neurotrophic factor, fibroblast growth factor-2 and Arc, known to suppress the apoptosis in the central nervous system, were significantly upregulated upon administration of nilvadipine. The present study indicates that misregulation of Rho phosphorylation may be involved as an important step in retinal degeneration of P23H and administration of nilvadipine may be a potential therapeutic agent for the retinal degenerations.

A case of superior segmental optic hypoplasia accompanied by a glaucomatous optic neuropathy.

This is the first case report of a bilateral superior segmental optic hypoplasia (SSOH) accompanied by a glaucomatous optic neuropathy (GON). A 47-year-old man incidentally diagnosed as having bilateral SSOH, simultaneously disclosed glaucomatous optic disc appearances, including enlargements of the cup of the optic nerve heads and a thinning of the infero-temporal neuroretinal rim with laminar dot sign accompanied by a retinal nerve fiber layer (RNFL) local defect of infero-temporal region in the right eye. The visual field examination revealed that the corresponding nasal step, arcuate scotoma and RNFLfield defects in the right eye.

Changes in intraocular indocyanine green concentrations during macular hole surgery.

PURPOSE: To report changes in intraocular indocyanine green (ICG) concentration which was used for visualization of the internal limiting membrane (ILM) during vitreous surgery on 3 eyes of 3 patients with full-thickness macular hole. METHODS: During intraocular surgery with ICG-assisted ILM peeling, aliquots of intraocular fluid obtained at 1 min after ICG administration (0.5 ml of 5 mg/ml ICG solution), before and after ILM peeling, and before and after fluid/gas exchange were subjected to spectrophotometric analysis. RESULTS: Intravitreous concentrations of ICG were dramatically reduced to approximately 0.4 microg/ml during the surgery for macular hole in all cases. CONCLUSION: The present study indicates that the levels of remaining intraocular ICG concentrations after macular hole surgery are much lower than the levels that induced retinal toxicity in experimental models.

Prolonged rhodopsin phosphorylation in light-induced retinal degeneration in rat models.

PURPOSE: The effects of various light-induced stresses on the retina were examined in the retinal degenerative rat model. METHODS: Retinal morphology and electroretinograms (ERGs) were analyzed after application of light-induced stress of several intensities (650, 1300, 2500, or 5000 lux). For evaluation of rhodopsin (Rho) function, the kinetics of Rho regeneration and dephosphorylation were studied by spectrophotometric analysis and immunofluorescence labeling with antibodies specifically directed toward the phosphorylated residues (334)Ser and (338)Ser in the C terminus of Rho. Retinal cGMP concentration was determined by ELISA. Expression levels of neurotrophic factors (FGF2, brain-derived neurotrophic factor [BDNF], platelet-derived growth factor [PDGF], and ciliary neurotrophic factor [CNTF]) were evaluated quantitatively by RT-PCR. RESULTS: Light intensity-dependent deterioration of ERG responses and thinning of the retinal outer nuclear layer were observed in wild-type and Royal College of Surgeons (RCS) rat retinas. Under dark adaptation after light-induced stress, the kinetics of Rho regeneration were not different between wild-type and RCS rat retinas. Rho dephosphorylation at (334)Ser and (338)Ser was extremely delayed in RCS rat retinas compared with wild-type without light-induced stress, but Rho dephosphorylation at those sites became slower in both RCS and wild-type rat retinas. In terms of expression of neurotrophic factors, almost no significant changes were observed between the animals after light-induced stress. CONCLUSIONS: The present study indicates that light-induced stress causes intensity-dependent deterioration in retinal function and morphology in wild-type and RCS rat retinas. Disruption of the phototransduction cascade resulting from slower kinetics of Rho dephosphorylation appears to be involved in retinal degeneration.

A case of normal tension glaucoma associated with Buerger's disease.

Open angle glaucoma, a slowly progressive optic atrophy, is clinically characterized by visual field defects corresponding to excavation of the optic disc, called glaucomatous cupping. Open angle glaucoma is further divided into primary open angle glaucoma caused by elevated intraocular pressure (higher than the normal limit of 21 mmHg), and normal tension glaucoma, in which intraocular pressure is in the normal range. Here we report a case of normal tension glaucoma associated with Buerger's disease, also known as thromboangiitis obliterans, which causes systemic blood flow disturbance. A 66-year-old man suffering from Buerger's disease for 10 years was diagnosed as having branch retinal artery occlusion in his left eye. He was referred to our clinic due to progressive visual field disturbance in that eye. Ophthalmologic examinations revealed occlusion in the infero-temporal retinal artery in the left eye, and glaucomatous cupping, normal intraocular pressure, retinal vessel tortuosity and retinal arteriosclerosis in both eyes. Visual field examination revealed decreased retinal sensitivities in the areas within the visual field arches above and below fixation from the blind spot to the median raphe, corresponding to the arcuate retinal nerve fibers comprising the Bjerrum areas and the area corresponding to the retinal artery occlusion. Buerger's disease is characterized by the development of segmental thrombotic occlusions and vasospasm of the medium and small arteries. Our case suggests that the blood flow disturbance due to arteriosclerosis, thrombotic occlusions and vasospasm associated with Buerger's disease might affect the ophthalmic circulation system, thereby contributing to the etiology of normal tension glaucoma.

Effects of antiglaucoma drops on MMP and TIMP balance in conjunctival and subconjunctival tissue.

PURPOSE: To study the effects of antiglaucoma drugs on metabolism within the extracellular matrix (ECM) of the ocular surface, including corneal, conjunctival, and subconjunctival tissue. METHOD: Several antiglaucoma drugs--including beta-blockers, alpha/beta-blockers, alpha1-blocker, alpha2-agonist, and prostaglandin derivative-were topically administrated to rat eyes daily for 2 weeks or were incubated with human corneal cells or human fibroblasts for 72 hours. Thereafter, expression and enzymatic activity of the matrix metalloproteinases (MMPs), a group of enzymes proteolyzing ECM and their inhibitors, called tissue inhibitors of metalloproteinase (TIMPs), were evaluated. RESULTS: Quantitative RT-PCR revealed significantly upregulated and downregulated expression of MMPs and TIMPs, respectively, in rat conjunctival and subconjunctival tissue on the administration of alpha/beta-blockers, alpha1-blocker, alpha2-agonist, and prostaglandin derivative, suggesting that these drugs may enhance ECM degradation. However, in contrast, beta-blocker administration caused reverse effects--that is, upregulation and downregulation of TIMPs and MMPs, respectively. Enzymatic activity of MMPs in rat conjunctival and subconjunctival tissue analyzed by biochemical assay and zymography was markedly enhanced on the administration of alpha/beta-blockers, alpha1-blocker, alpha2-agonist, and prostaglandin derivative, but not of beta-blockers. Similar effects of these antiglaucoma drugs were observed in cultured human corneal cells and human fibroblast cells. CONCLUSIONS: The present experimental observations suggest that some alpha/beta-blockers, alpha1-blocker, alpha2-agonist, and prostaglandin derivative stimulate ECM degradation of ocular surface tissue by modulating the balance between MMPs and TIMPs.

Study of effects of antiglaucoma eye drops on N-methyl-D-aspartate-induced retinal damage.

PURPOSE: To study the effects of antiglaucoma eye drops on N-methyl-D-aspartate (NMDA)-induced retinal damage. METHODS: Several antiglaucoma eye drops, beta-blockers, alpha/beta-blockers, an alpha1-blocker, an alpha2-agonist, and a prostaglandin derivative, were topically administrated to NMDA-treated rat eyes daily for 2 weeks, and the retinal thickness, the number of retrograde-labeled retinal ganglion cells (RGCs), and the results of a cDNA microarray analysis were studied. RESULTS: Intravitreal administration of NMDA caused a significant decrease in the thickness of the retinal layers and induced upregulation of glial fibrillary acidic protein (GFAP). Topical administration of beta-blockers (timolol, betaxolol, and carteolol) and a prostaglandin derivative (latanoprost) showed almost no significant effects on retinal thickness, the number of RGCs, or expression of GFAP. In contrast, the alpha/beta-blockers (nipradilol and levobunolol), the alpha1-blocker (bunazosin HCl), and the alpha2-agonist (brimonidine) showed preservation effects on retinal thickness and the number of RGCs, and marked suppression of NMDA-induced upregulation of GFAP. Among 1101 genes related to cellular regulatory mechanisms, the expression of two genes, both for insulin-like growth factors, (IGF-1) and ErbB3, was altered upon administration of the alpha/beta-blockers, the alpha1-blocker, and the alpha2-agonist. CONCLUSION: Our present study suggests that modulations of the alpha-adrenergic receptor, alpha1-blocking and alpha2-stimulation, by antiglaucoma eye drops may cause beneficial effects on NMDA-induced retinal damage in the rat.

Effects of matrix metalloproteinase-3 gene transfer by electroporation in glaucoma filter surgery.

To develop gene therapy that can be applied to glaucoma-filtering surgery, we studied effects of transfection of matrix metalloproteinase-3 (MMP-3) cDNA into rabbit conjunctiva by electroporation (EP) on changes of intraocular pressure (IOP) and bleb formation after glaucoma filtering surgery. pTracer-CMV2 vector containing MMP-3 cDNA was transfected into rabbit conjunctiva by EP and MMP-3 expression was studied by reverse transcription (RT)-PCR, zymography and western blot analysis. Three days after the EP transfection of MMP-3 cDNA or vector alone into rabbit conjunctiva, trabeculectomy was performed at the place of transfection in the presence or absence of 0.04% mitomycin C (MMC). Then changes in IOPs and bleb formation were compared with each other. Expression of MMP-3 was detected in conjunctiva until 30 days after transfection by EP. Trabeculectomy following MMP-3 transfection caused significantly longer survival of filtering bleb and decreased levels of IOP in comparison with controls (trabeculectomy alone or trabeculectomy following vector transfection), and these levels were almost identical to those of trabeculectomy with MMC. The present study indicates that EP is effective to transfect some genes that promote the filtering bleb formation in glaucoma surgery, such as MMP-3 gene, and this may be potentially applicable to glaucoma-filtering surgery in glaucoma patients.

Clinical and immunologic aspects of cancer-associated retinopathy.

PURPOSE: To report clinical and immunologic aspects of cancer-associated retinopathy (CAR). DESIGN: Observational consecutive case series. METHODS: A retrospective review was made of 18 consecutive patients with cancer-associated retinopathy who had antiretinal antibody determination by Western blot testing. RESULTS: Clinically, a variety of ophthalmic observations including electroretinography impairment, retinal vessel narrowing, deterioration of visual acuity, visual field changes, and uveitis were frequently observed. As retinal autoantigens in the 18 cases, recoverin was found in all 18 cases (100%), heat shock cognate protein 70 (HSC70) was found in six cases (33%), and other proteins were found in four cases (20%). These antibodies were detected in only 60% of the patients at the initial examination, however, and then became increasingly apparent on the subsequent testing that was performed three times on serum samples obtained sequentially during the following months. CONCLUSION: For diagnosis of cancer-associated retinopathy, the presence of serum autoantibody toward recoverin is essentially required in addition to the characteristic clinical aspects noted above.

Clinicopathological features of gastric cancer cases and aberrantly expressed recoverin.

Cancer-associated retinopathy (CAR) is an ocular manifestation of a paraneoplastic syndrome whereby immunological reactions toward recoverin, a retina-specific calcium binding protein, and other retinal antigens aberrantly expressed in tumor cells lead to the degeneration of retinal photoreceptor cells. Recently we reported that aberrant expression of recoverin was identified in more than 50% of tumor cells and their cell lines from several kinds of cancers, including gastric cancer, lung carcinoma, and other cancers. To elucidate the clinicopathological significance of aberrantly expressed recoverin in cancer cells, we performed immunocytochemical analysis using a monoclonal antibody against human recoverin. Within 18 patients with different clinical stages (I-IV) of gastric cancer, the aberrant expression of recoverin in tumor cells was recognized in 6 out of 18 patients (2 out of 2 stage IA, 1 out of 1 stage IB, 2 out of 3 stage II, 0 out of 7 stage IIIA, 0 out of 1 stage IIIB, and 1 out 4 stage IV). The present data are consistent in part with the previous observations that recoverin-expressing cancer cells induced tumor immunity and provide a favorable prognosis for primary cancer in CAR patients.

Study of drug effects of calcium channel blockers on retinal degeneration of rd mouse.

In the present study, we studied drug effects of Ca(2+) antagonists on the retinal degeneration of rd mouse to evaluate their efficacy. Several kinds of Ca(2+) antagonists, diltiazem, nicardipine, nilvadipine or nifedipine were administrated intraperitoneally and thereafter retinal morphology and functions were analyzed. In addition, we performed DNA microarray analysis both in nilvadipine treated and control retinas to understand their drug effects at molecular levels. We found that nilvadipine caused significant preservation of retinal thickness in rd mouse during the initial stage of the retinal degeneration, and nicardipine showed also significant but lesser preservation than nilvadipine. However, we recognized no preservation effects of diltiazem and nifedipine. In the total 3774 genes, the expressions of 27 genes were altered upon administration of nilvadipine, including several genes related to the apoptotic pathway, neuro-survival factor, Ca(2+) metabolisms, and other mechanisms. It is suggested that some types of Ca(2+) channel blockers, such as nilvadipine and nicardipine, are able to preserve photoreceptor cells in rd mouse and can potentially be used to treat some RP patients.

Low expression of rhodopsin kinase in pineal gland in Royal College of Surgeons rat.

The Royal College of Surgeons (RCS) rat has been extensively characterized as a model for inherited retinal dystrophy such as retinitis pigmentosa. We have found that significantly low levels of expression of rhodopsin kinase (RK) and alphaA-crystallin may be involved in the pathogenesis of retinal degeneration in the RCS rat (Invest Ophthalmol Vis Sci. 1999,40:2788-2794). In the present study, we examined the expression of photoreceptor specific proteins in the pineal gland (PG) including rhodopsin kinase (RK), arrestin and recoverin, which are known to be commonly present in both photoreceptor and PG, in order to elucidate the pathological relationship between retina and PG during retinal degeneration. Among these proteins, RK expression was significantly decreased with advancing age (3-5 weeks old) in RCS rat. However, in contrast, arrestin expression in RCS PG was comparable with control PG and no expressions of recoverin and other G-protein coupled receptor kinases (GRKs 2, 5 and 6) were detected in RCS PG during 3-5 weeks of age. By administration of nilvadipine, an effective Ca2+ antagonist that was shown to preserve RCS retinal degeneration, RK expression was significantly enhanced.