LINE-1 hypomethylation, increased retrotransposition and tumor-specific insertion in upper gastrointestinal cancer.

The long interspersed nuclear element-1 (LINE-1) retrotransposons are a major family of mobile genetic elements, comprising approximately 17% of the human genome. The methylation state of LINE-1 is often used as an indicator of global DNA methylation levels and it regulates the retrotransposition and somatic insertion of the genetic element. We have previously reported the significant relationship between LINE-1 hypomethylation and poor prognosis in upper gastrointestinal (GI) cancers. However, the causal relationships between LINE-1 hypomethylation, retrotransposition, and tumor-specific insertion in upper GI cancers remain unknown. We used bisulfite-pyrosequencing and quantitative real-time PCR to verify LINE-1 methylation and copy number in tissue samples of 101 patients with esophageal and 103 patients with gastric cancer. Furthermore, we analyzed the LINE-1 retrotransposition profile with an originally developed L1Hs-seq. In tumor samples, LINE-1 methylation levels were significantly lower than non-tumor controls, while LINE-1 copy numbers were markedly increased. As such, there was a significant inverse correlation between the LINE-1 methylation level and copy number in tumor tissues, with lower LINE-1 methylation levels corresponding to higher LINE-1 copy numbers. Of particular importance is that somatic LINE-1 insertions were more numerous in tumor than normal tissues. Furthermore, we observed that LINE-1 was inserted evenly across all chromosomes, and most often within genomic regions associated with tumor-suppressive genes. LINE-1 hypomethylation in upper GI cancers is related to increased LINE-1 retrotransposition and tumor-specific insertion events, which may collectively contribute to the acquisition of aggressive tumor features through the inactivation of tumor-suppressive genes.

Statins exert anti-growth effects by suppressing YAP/TAZ expressions via JNK signal activation and eliminate the immune suppression by downregulating PD-L1 expression in pancreatic cancer.

Statins are cholesterol-lowering agents that act as inhibitors of 3-hydroxy-3-methyl-glutaryl-coenzymeA (HMG CoA) reductase. Recently, statins have received a lot of attention, especially regarding how statins act on the immune system. Here, the clinical impact of statin intake was examined in patients with resected pancreatic cancer, and the underlying mechanisms were investigated in vitro and in vivo. We found that statin intake was associated with favorable prognostic outcomes in patients with resectable pancreatic cancer. Statins, especially lipophilic statins, exert anti-proliferative effects on pancreatic cancer cells in vitro (simvastatin > fluvastatin > atorvastatin > rosuvastatin > pravastatin). Simvastatin had an anti-proliferative effect on pancreatic cancer cells with decreased the yes-associated protein (YAP)/PDZ-binding motif (TAZ) expression by activating the JNK pathway, and simvastatin treatment with oxaliplatin revealed additive anti-growth effects. Furthermore, lipophilic and hydrophilic statins suppressed programmed cell death ligand 1 (PD-L1) expression by downregulating TAZ. Simvastatin treatment with an anti-PD-1 drug (BP0273) provided immediate anti-growth effects compared to controls, such as anti-PD-1 only and simvastatin only, and suppressed progressive disease during the early period of anti-PD-1 treatment in vivo. In conclusion, Statins display two distinct anti-cancer effects (direct anti-growth effect and elimination of immune suppression by downregulating PD-L1 expression) by targeting YAP/TAZ expression.

Twenty-Four-Hour Variation of Intraocular Pressure in Primary Open-Angle Glaucoma Treated with Triple Eye Drops.

Objectives. To evaluate 24-hour intraocular pressure (IOP) variation in patients with primary open-angle glaucoma (POAG) treated with triple eye drops. Subjects and Methods. The IOP was measured in 74 eyes in 74 POAG patients (seated) on triple therapy (PG analogue, ß-blocker, carbonic anhydrase inhibitor) at about every 3 hours. Results. The peak IOP was 13.5 ± 3.1 at 1:00, and the trough IOP was at 12.6 ± 2.4 mmHg at 7:00. The IOP at 7:00 was significantly lower than that at 10:00, 1:00, and 3:00 (p < 0.05). Based on the time of the peak IOP, we classified the patients into two groups: diurnal (28 eyes) and nocturnal types (37 eyes). There was significant difference at the spherical equivalent between diurnal and nocturnal types (p = 0.014). To assess the influence of reflective error, we conducted subanalysis for two groups: high myopic (26 eyes, ≤-6D) and low/nonmyopic (24 eyes, ≥-2D) groups. In the low/nonmyopia group, the IOP was significantly higher at 1:00 and 3:00 than at 13:00, 16:00, and 7: 00 (p < 0.05). Conclusion. The mean of IOP elevated outside of clinic hour in the POAG patients on triple therapy. The low/nonmyopia patient should be carefully treated because the IOP of the patients at night elevated significantly.

Tracing behavior of endothelial cells promotes vascular network formation.

The in vitro formation of network structures derived from endothelial cells in grafts before transplantation contributes to earlier engraftment. In a previous study, endothelial cells migrated to form a net-shaped structure in co-culture. However, the specific network formation behavior of endothelial cells during migration remains unclear. In this study, we demonstrated the tracing behavior and cell cycle of endothelial cells using Fucci-labeled (Fluorescent Ubiquitination-based Cell Cycle Indicator) endothelial cells. Here, we observed the co-culture of Fucci-labeled human umbilical vein endothelial cells (HUVECs) together with normal human dermal fibroblasts (NHDFs) using time-lapse imaging and analyzed by multicellular concurrent tracking. In the G0/G1 period, HUVECs migrate faster than in the S/G2/M period, because G0/G1 is the mobile phase and S/G2/M is the proliferation phase in the cell cycle. When HUVECs are co-cultured, they tend to move randomly until they find existing tracks that they then follow to form clusters. Extracellular matrix (ECM) staining showed that collagen IV, laminin and thrombospondin deposited in accordance with endothelial cell networks. Therefore the HUVECs may migrate on the secreted ECM and exhibit tracing behavior, where the HUVECs migrate toward each other. These results suggested that ECM and a cell phase contributed to form a network by accelerating cell migration.

Effective recovery of highly purified CD326(+) tumor cells from lavage fluid of patients treated with a novel cell-free and concentrated ascites reinfusion therapy (KM-CART).

For the production of tumor-specific vaccines, including dendritic cell (DC) vaccines, the tumor cells themselves are an ideal source. Floating tumor cells in the ascites fluid from patients with malignant ascites are a good candidate source, but it is not easy to obtain pure tumor cells from ascites because of various types of cell contamination as well as protein aggregates. We here report an effective method to recover pure tumor cells from malignant ascites. We used lavage fluid from 13 patients with malignant ascites who were treated with modified cell-free and concentrated ascites reinfusion therapy (KM-CART). Cellular components were separated from the lavage fluid by centrifugation, enzymatic digestion and hemolysis. Tumor cells were purified by depleting CD45(+) leukocytes with antibody-conjugated magnetic beads. The tumor cell lysate was extracted by freeze-and-thaw cycles. The mean obtained total cell number was 7.50 × 10(7) cells (range 4.40 × 10(6)-2.48 × 10(8) cells). From this fraction, 6.39 × 10(6) (range 3.23 × 10(5)-2.53 × 10(7)) CD45(-) cells were collected, and the tumor cell purity was over 80 % defined as CD45(-)CD326(+). A sufficient amount of tumor lysate, average  = 2416 µg (range 25-8743 µg), was extracted from CD45(-)CD326(+) tumor cells. We here established an effective method to produce highly purified tumor cells from KM-CART lavage fluid. The clinical feasibility of this simple preparation method for generating tumor lysate should be examined in clinical studies of DC vaccines.

Republication: Two Premature Neonates of Congenital Syphilis with Severe Clinical Manifestations.

Congenital syphilis (CS) is a public health burden in both developing and developed countries. We report two cases of CS in premature neonates with severe clinical manifestations; Patient 1 (gestational age 31 weeks, birth weight 1423 g) had disseminated idiopathic coagulation (DIC) while Patient 2 (gestational age 34 weeks and 6 days, birth weight 2299 g) had refractory syphilitic meningitis. Their mothers were single and had neither received antenatal care nor undergone syphilis screening. Both neonates were delivered via an emergency cesarean section and had birth asphyxia and transient tachypnea of newborn. Physical examination revealed massive hepatosplenomegaly. Laboratory testing of maternal and neonatal blood showed increased rapid plasma reagin (RPR) titer and positive Treponema pallidum hemagglutination assay. Diagnosis of CS was further supported by a positive IgM fluorescent treponemal antibody absorption test and large amounts of T. pallidum spirochetes detected in the placenta. Each neonate was initially treated with ampicillin and cefotaxime for early bacterial sepsis/meningitis that coexisted with CS. Patient 1 received fresh frozen plasma and antithrombin III to treat DIC. Patient 2 experienced a relapse of CS during initial antibiotic treatment, necessitating parenteral penicillin G. Treatment was effective in both neonates, as shown by reductions in RPR. Monitoring of growth and neurological development through to age 4 showed no evidence of apparent delay or complications. Without adequate antenatal care and maternal screening tests for infection, CS is difficult for non-specialists to diagnose at birth, because the clinical manifestations are similar to those of neonatal sepsis and meningitis. Ampicillin was insufficient for treating CS and penicillin G was necessary.

[Correlation of intraocular pressure variation after visual field examination with 24-hour intraocular pressure variations in primary open-angle glaucoma].

OBJECTIVES: We retrospectively examined intraocular pressure variations after visual field examination in primary open angle glaucoma (POAG), together with its influencing factors and its association with 24-hour intraocular pressure variations. SUBJECTS AND METHODS: Subjects were 94 eyes (52 POAG patients) subjected to measurements of 24-hour intraocular pressure and of changes in intraocular pressure after visual field examination using a Humphrey Visual Field Analyzer. Subjects were classified into three groups according to the magnitude of variation (large, intermediate and small), and 24-hour intraocular pressure variations were compared among the three groups. Factors influencing intraocular pressure variations after visual field examination and those associated with the large variation group were investigated. RESULTS: Average intraocular pressure variation after visual field examination was -0.28 ± 1.90 (range - 6.0(-) + 5.0) mmHg. No significant influencing factors were identified. The intraocular pressure at 3 a.m. was significantly higher in the large variation group than other two groups (p < 0.001). Central corneal thickness was correlated with the large variation group (odds ratio = 1.04; 95% confidence interval, 1.01-1.07 ; p = 0.02). CONCLUSION: No particular tendencies in intraocular pressure variations were found after visual field examination. Increases in intraocular pressure during the night might be associated with large intraocular pressure variations after visual field examination.

Intraocular pressure change over a habitual 24-hour period after changing posture or drinking water and related factors in normal tension glaucoma.

PURPOSE: We investigated the correlation between 24-hour IOP in the habitual (sitting during day and supine during night) position (H24h-IOP) and IOP after a postural-change test (PCT-IOP) and a water-drinking test (WDT-IOP). We also investigated ocular and systemic factors related with them in patients with normal tension glaucoma (NTG). METHODS: Japanese NTG patients underwent H24h-IOP, PCT-IOP, and WDT-IOP measurements during a 24-hour period. Correlations among H24h-IOP, PCT-IOP, and WDT-IOP, and contributing ocular/systemic factors were investigated using regression analysis. RESULTS: There were 33 patients included. Peak H24h-IOP correlated positively with peak PCT-IOP and peak WDT-IOP (estimate = 0.422 and 0.419, P ≤ 0.010), and peak PCT-IOP with WDT-IOP (0.44, P = 0.002). Peak H24h-IOP correlated with refraction (0.36, P = 0.048) and negatively with the mean deviation (MD, -0.066, P = 0.031). MD and baseline IOP (the mean of H24h-IOP) correlated negatively with the H24h-IOP fluctuation (-0.058 and -0.58, P ≤ 0.050). Refraction, baseline IOP, mean blood pressure (mBP), and body mass index (BMI) correlated with peak PCT-IOP (0.23, 0.52, 0.097, and 0.32, respectively, P ≤ 0.038). PCT-IOP difference correlated with refraction and mBP (0.31 and 0.093, P ≤ 0.016) and negatively with age (-0.069, P = 0.003). Central corneal thickness, baseline IOP, age, and BMI correlated with peak WDT-IOP (0.030, 0.40, 0.088, and 0.26, P ≤ 0.050). Age and BMI correlated with WDT-IOP difference (0.086 and 0.20, P < 0.032). CONCLUSIONS: Positive correlation was found among the peaks of H24h-, PCT-, and WDT-IOP. A worse visual field was associated with higher peak and greater fluctuation of H24h-IOP in NTG. Several ocular/systemic factors were important in interpreting H24h-, PCT-, and WDT-IOP.

Association of HK2 and NCK2 with normal tension glaucoma in the Japanese population.

Although family studies and genome-wide association studies have shown that genetic factors play a role in glaucoma, it has been difficult to identify the specific genetic variants involved. We tested 669 single nucleotide polymorphisms (SNPs) from the region of chromosome 2 that includes the GLC1B glaucoma locus for association with primary open-angle glaucoma (POAG) and normal tension glaucoma (NTG) in the Japanese population. We performed a two-stage case-control study. The first cohort consisted of 123 POAG cases, 121 NTG cases and 120 controls: the second cohort consisted of 187 POAG cases, 286 NTG cases, and 271 controls. Out of six SNPs showing significant association with POAG in the first round screening, seven SNPs were tested in the second round. Rs678350 in the HK2 gene coding sequence showed significant allelic (p=0.0027 in Stage Two, 2.7XE-4 in meta-analysis) association with POAG, and significant allelic (p=4.7XE-4 in Stage Two, 1.0XE-5 in meta-analysis) association with NTG. Although alleles in the TMEM182 gene did not show significant association with glaucoma in the second round, subjects with the A/A allele in TMEM182 rs869833 showed worse visual field mean deviation (p=0.01). Even though rs2033008 in the NCK2 gene coding sequence did not show significant association in the first round, it had previously shown association with NTG so it was tested for association with NTG in round 2 (p=0.0053 in Stage Two). Immunohistochemistry showed that both HK2 and NCK2 are expressed in the retinal ganglion cell layer. Once multi-testing was taken into account, only HK2 showed significant association with POAG and NTG in Stage Two. Our data also support previous reports of NCK2 association with NTG, and raise questions about what role TMEM182 might play in phenotypic variability. Our data suggest that HK2 may play an important role in NTG in the Japanese population.

Association of Toll-like receptor 4 gene polymorphisms in Japanese subjects with primary open-angle, normal-tension, and exfoliation glaucoma.

PURPOSE: To determine whether polymorphisms in the Toll-like receptor 4 (TLR4) gene are associated with primary open-angle glaucoma (POAG), normal-tension glaucoma (NTG), and exfoliation glaucoma (XFG) in Japanese individuals. DESIGN: Genetic association study. SETTING: Multicenter study. STUDY POPULATION: One hundred eighty-four unrelated Japanese patients with POAG, 365 unrelated patients with NTG, and 109 unrelated patients with XFG from 5 hospitals. PROCEDURES: Genomic DNA was extracted from leukocytes of the peripheral blood, and 8 polymorphisms in the TLR4 genes were amplified by polymerase chain reaction (PCR) and directly sequenced. Allele and genotype frequencies and the inferred haplotypes were estimated. MAIN OUTCOME MEASURES: Differences in allele and genotype frequencies and haplotypes between subjects with POAG, NTG, and XFG. RESULTS: The allele frequency of rs2149356 of the TLR4 gene in the POAG, NTG, and XFG groups was the most significantly different from that of the control group (minor allele frequency 0.446, 0.395, 0.404, vs 0.308; P = .000058, P = .0030, and P = .015). The allele frequencies of the 5 TLR4 SNPs were higher in all of the glaucoma groups than that in the control group. The statistics of genotypes of TLR4 were approximately the same for all allele frequencies. The haplotypic frequencies with Tag SNPs studied earlier showed that only POAG was statistically significant. Other haplotypes, such as rs10759930, rs1927914, rs1927911, and rs2149356, had higher statistical significance (overall P = .00078 in POAG, overall P = .018 in NTG, and overall P = .014 in XFG). CONCLUSIONS: This study demonstrated that TLR4 polymorphisms are associated with NTG in the Japanese, and they also play a role in the pathogenesis of POAG and XFG.

Common variants on chromosome 9p21 are associated with normal tension glaucoma.

Although intraocular pressure (IOP) is the most definitive cause of glaucoma, a subtype of open angle glaucoma (OAG) termed normal tension glaucoma (NTG), which occurs in spite of normal IOP, accounts for a large part of glaucoma cases, especially in Japan. To find common genetic variants contributing to NTG in Japanese patients, we conducted a genome-wide association study (GWAS). We performed the first screening for 531,009 autosomal SNPs with a discovery cohort of 286 cases and 557 controls, and then a second screening for the top 30 suggestive loci in an independent cohort of 183 cases and 514 controls. Our findings identified a significantly associated SNP; rs523096 [combined p-value = 7.40× 10(-8), odds ratio (OR)= 2.00 with 95% confidence interval (CI) 1.55-2.58] located 10 kbp upstream of CDKN2B on chromosome 9p21. Moreover, analysis of another independent case-control set successfully replicated the results of the screening studies (combined values of all 3 stages p = 4.96 × 10(-11), OR= 2.13 with 95% CI 1.69-2.68). The SNPs near rs523096 were recently reported to be associated with OAG associated with elevated IOP in primary open-angle glaucoma (POAG), the predominant subtype of glaucoma in Caucasian populations. Our results revealed that the 9p21 locus is also associated with NTG in Japanese. In addition, we identified SNPs more strongly associated with NTG.

Creation of mouse embryonic stem cell-derived cardiac cell sheets.

Research on heart tissue engineering is an exciting and promising area. Although we previously developed bioengineered myocardium using cell sheet-based tissue engineering technologies, the issue of appropriate cell sources remained unresolved. In the present study, we created cell sheets of mouse embryonic stem (ES) cell-derived cardiomyocytes after expansion in three-dimensional stirred suspension cultures. Serial treatment of the suspension cultures with noggin and granulocyte colony-stimulating factor significantly increased the number of cardiomyocytes by more than fourfold compared with untreated cultures. After drug selection for ES cells expressing the neomycin-resistance gene under the control of the α-myosin heavy chain promoter, almost all of the cells showed spontaneous beating and expressed several cardiac contractive proteins in a fine striated pattern. When ES-derived cardiomyocytes alone were seeded onto temperature-responsive culture dishes, cell sheets were not created, whereas cocultures with cardiac fibroblasts promoted cell sheet formation. The cardiomyocytes in the cell sheets beat spontaneously and synchronously, and expressed connexin 43 at the edge of adjacent cardiomyocytes. Furthermore, when the extracellular action potential was recorded, unidirectional action potential propagation was observed. The present findings suggest that stirred suspension cultures with appropriate growth factors are capable of producing cardiomyocytes effectively and easily, and that ES-derived cardiac cell sheets may be a promising tool for the development of bioengineered myocardium.

Effect of carteolol hydrochloride on 24-hour variation of intraocular pressure in normal-tension glaucoma.

PURPOSE: To evaluate the effects of carteolol hydrochloride (carteolol) on 24-h variations in intraocular pressure (IOP) in patients with normal-tension glaucoma (NTG). METHODS: Twelve patients with NTG were treated with carteolol 2% solution for >or=8 weeks; their pretreatment 24-h IOP variations, blood pressure (BP), and pulse rate (PR) were compared with those measured after the treatment period. RESULTS: Daytime IOP (at 07:00, 10:00, 13:00, and 16:00), maximum IOP, and the mean 24-h IOP were significantly reduced after treatment, as was the 24-h IOP range. Systolic BP in the morning and both systolic and diastolic BP in the afternoon were significantly decreased by the treatment, whereas no significant change of PR was observed. CONCLUSIONS: Carteolol had no effect on nocturnal IOP but significantly helped reduce daytime IOP, maximum IOP, mean 24-h IOP, and the 24-h IOP range. The drug exerted no statistically significant effect on the PR.

Interleukin-10 and interleukin-6 in aqueous humor during treatment of vitreoretinal lymphoma with intravitreally injected methotrexate.

OBJECTIVE: To evaluate the effectiveness of measuring anterior chamber interleukin (IL)-10 and IL-6 concentrations during treatment of vitreoretinal lymphoma with intravitreally injected methotrexate. METHODS: Two patients (3 eyes) underwent repeated weekly intravitreal injections of methotrexate (400 microg of methotrexate/50 microl) for 6 weeks. We monitored the IL-10 and IL-6 concentrations in the anterior chamber and the IL-10/IL-6 ratio at baseline and at the last injections using a standard enzyme-linked immunoassay. RESULTS: Repeated intravitreal injections of methotrexate reduced the vitreous opacity. The IL-10/IL-6 ratio decreased consistently in conjunction with clearance of the vitreous opacity in all 3 eyes, from 39.2 to 0.02, 1.67 to 0.03 and 3.05 to 0.159, respectively. CONCLUSIONS: A decrease in the IL-10/IL-6 ratio seems to be related to a decrease in vitreous opacity. Measuring the IL-10/IL-6 ratio in the anterior chamber may be useful to evaluate the effect of treatment of vitreoretinal lymphoma.

[The relationship between difficulty in daily living and binocular visual field in patients with glaucoma].

PURPOSE: To evaluate the relationship between difficulty in daily living and binocular visual field in patients with glaucoma. SUBJECTS AND METHODS: We examined 144 patients with glaucoma who had bilateral visual field disturbances. The mean age was 68 +/- 13 years. Difficulty in daily living was assessed using a questionnaire consisting of 10 questions on daily activities. Each response was rated on a scale of 1 to 4: with a score of 4 for "no difficulty", 3 for "some difficulty", 2 for "great difficulty" and 1 for "cannot manage at all". The binocular visual field was assessed using the Esterman visual field test and the Esterman score was calculated. The relationship between difficulty in daily living and Esterman score was evaluated. RESULTS: The mean total score for difficulty in daily living was 34 +/- 7 (range; 13-40). The mean Esterman score was 79 +/- 24 (range; 0-100). The total score for difficulty in daily living correlates positively with the Esterman score (r2 = 0.48, p < 0.0001). CONCLUSION: The present study indicates that the Esterman score is a good predictor of difficulty daily living.

[Correlation of age and intraocular pressure with visual field damage in patients with normal-tension glaucoma].

OBJECTIVE: We investigated the correlation of age and intraocular pressure (IOP) with visual field damage in patients with normal-tension glaucoma (NTG) and asymmetric visual field defects. METHODS: A total of 88 NTG patients (176 eyes) were investigated. Patients in whom visual field defects were more severe in the eye with higher IOP than in the fellow eye were grouped as IOP-visual field-concordant patients, and the others as IOP-visual field-discordant patients. Mean 24-hour IOP was used as an indicator of IOP, and mean deviation (MD) as an indicator of visual field defects. Patients were also compared by age tertile. RESULTS: The IOP-visual field-concordant group comprised 53 of the 88 patients (60.2%), among whom the >70-years tertile had significantly greater absolute values for MD asymmetry. In contrast, no significant differences in asymmetry by age tertile were seen in the IOP-visual field-discordant group. CONCLUSION: Elevated IOP is a cause of visual field defects in many NTG patients. Among IOP-visual field-concordant patients, particular attention is required to visual field changes in the eyes of patients with higher IOP.

Flow cytometric evaluation of synergistic pro-apoptotic effects of statins and clofibrates in IM-9 human lymphoblasts.

1. In the present study, we evaluated fibrate-mediated potentiation of statin-induced apoptosis in IM-9 human lymphoblasts. 2. The pro-apoptotic effects of statin and fibrate were measured by flow cytometry with biotin-annexin V, followed by addition of avidin-fluorescein isothiocyanate and propidium iodide. Apoptosis was confirmed using karyopyknotic staining, as well as detection of DNA fragmentation and caspase 3 activation. 3. Incubation of IM-9 cells with both 0.1 micromol/L cerivastatin and 200 micromol/L clofibrate had a synergistic effect compared with 0.1 micromol/L cerivastatin alone or 200 micromol/L clofibrate alone. The magnitude of apoptosis induced by various combinations of statins and clofibrate were as follows: cerivastatin (0.1 micromol/L) + clofibrate (200 micromol/L) > atorvastatin (0.1 micromol/L) + clofibrate (200 micromol/L) > pravastatin (100 micromol/L) + clofibrate (200 micromol/L). Other fibrates (bezafibrate and clinofibrate) did not show any synergistic effect. Furthermore, karyopyknotic staining, caspase 3 activation and DNA fragmentation demonstrated synergistic pro-apoptotic effects of statin and fibrate. 4. The results of the present study suggest that simultaneous treatment with statins and clofibrate could provide improved therapeutic efficacy in leukaemia patients.

Effect of concomitant use of latanoprost and brinzolamide on 24-hour variation of IOP in normal-tension glaucoma.

PURPOSE: To study the effect of the concomitant use of latanoprost and brinzolamide on the 24-hour variation in the intraocular pressure (IOP) in patients with normal-tension glaucoma (NTG). METHODS: We studied a total of 44 eyes from 22 NTG patients. Mean 24-hour IOP variation was determined after a washout period of > or =4 weeks. Latanoprost monotherapy was continued in both eyes for 8 weeks. Thereafter, patients were randomized to continue latanoprost monotherapy in 1 eye whereas brinzolamide was added as an adjunct to latanoprost therapy in the other eye. Eight weeks after the initiation of brinzolamide treatment, the 24-hour IOP variation was remeasured. IOP was measured in the sitting position 8 times daily using a Goldmann applanation tonometer before and after treatment. RESULTS: The eyes treated with latanoprost monotherapy and those treated with latanoprost and brinzolamide showed a significant decrease in IOP at all time points. Percent reductions in the diurnal mean IOP (mean IOP at 10 AM, 1 PM, and 4 PM) and in nocturnal mean IOP (mean IOP at 10 PM, 1 AM, and 3 AM) were significantly greater in the eyes treated with the combination of latanoprost and brinzolamide than those with latanoprost alone (diurnal mean IOP: latanoprost and brinzolamide=19.8%, latanoprost=14.1%, P<0.001; nocturnal mean IOP: latanoprost and brinzolamide=13.4%, latanoprost=10.0%, P<0.05). CONCLUSIONS: For the treatment of NTG, the combination of latanoprost and brinzolamide demonstrated additive effects in lowering IOP, not only during the day, but also at night.

[Reading performance in patients with central visual field disturbance due to glaucoma].

PURPOSE: To evaluate the relationship between central visual field disturbance due to glaucoma and reading performance. SUBJECTS AND METHODS: We evaluated the reading performance of 11 patients (14 eyes) with visual acuities of 1.0 and higher who had absolute scotoma within 3 degrees of the central visual field. The ages of the patients ranged from 37 to 77, with a median of 62 years. Maximum reading speed of texts printed horizontally and vertically was measured using MNREAD-J. Subjective reading difficulties were investigated orally. The relationship between the maximum reading speed of both horizontal and vertical texts, the subjective reading difficulties, the number of quadrants, and the position and continuity of absolute scotoma were evaluated. RESULTS: The mean maximum reading speed for vertical and horizontal texts by the 11 patients was 323 and 335 characters/minute(c/m), respectively, and was not significantly slower than that of normal eyes. In 2 of 14 eyes, a significant (p < 0.05) difference between maximum vertical and horizontal reading speeds was observed. In 3 of 14 eyes, absolute scotoma was distributed continuously in more than 2 adjacent quadrants, and these patients also had subjective reading difficulties. CONCLUSION: Reading difficulty is present in patients having absolute scotoma within 3 degrees of the central visual field when the scotoma involves more than 2 adjacent quadrants.

SNPs and interaction analyses of noelin 2, myocilin, and optineurin genes in Japanese patients with open-angle glaucoma.

PURPOSE: To evaluate the noelin 2 gene as a disease-causing factor for open-angle glaucoma (OAG) and the interactions between the noelin 2 (OLFM2), optineurin (OPTN), and myocilin (MYOC) genes. METHODS: OLFM2 was analyzed in 770 Japanese subjects including 215 patients with elevated intraocular pressure (IOP), 277 with normal IOP, 38 with juvenile open-angle glaucoma, and 240 control subjects. Two single-nucleotide polymorphisms (SNPs) in OPTN (c.412G-->A and c.603T-->A) and one SNP in MYOC (c.227G-->A) were examined. Single genes were investigated by univariate analysis and the gene-gene interactions by logistic regression analysis. Associations between genotypes and clinical characteristics at the time of diagnosis were examined. RESULTS: In OLFM2, 12 sequence variants were identified in 770 Japanese subjects. Arg144Gln (exon 4) was identified in two (0.3%) of the patients and in none of the control subjects. Combinations of OLFM2/317A and OPTN/412A or OLFM2/1281T and OPTN/412A were associated with patients with elevated IOP (P = 0.018 or P = 0.012, respectively). The combination of OLFM2/317G and OPTN/603A was significantly associated with elevated IOP (P = 0.018). No significant association was detected between SNPs in OLFM2 and in MYOC. Patients with normal IOP and with OLFM2/678A+OPTN/412G or OLFM2/1281C+OPTN/412G had significantly worse visual field scores (P = 0.022 or 0.030, respectively). CONCLUSIONS: The Arg144Gln mutation in OLFM2 is a possible disease-causing mutation in Japanese patients with OAG. Common polymorphisms in OLFM2 and OPTN may interactively contribute to the development of OAG, indicating a polygenic etiology.