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Purpose To investigate whether processing visual field (VF) measurements using a variational autoencoder (VAE) improves the structure–function relationship in glaucoma. Design Cross-sectional study. Participants The training data consisted of 82 433 VF measurements from 16 836 eyes. The testing dataset consisted of 117 eyes of 75 patients with open-angle glaucoma. Methods A VAE model to reconstruct the threshold of VF was developed using the training dataset. OCT and VF (Humphrey Field Analyzer 24-2, Swedish interactive threshold algorithm standard) measurements were carried out for all eyes in the testing dataset. Visual fields in the testing dataset then were reconstructed using the trained VAE. The structure–function relationship between the circumpapillary retinal nerve fiber layer (cpRNFL) thickness and VF sensitivity was investigated in each of twelve 30° segments of the optic disc (3 nasal sectors were merged). Similarly, the structure–function relationship was investigated using the VAE-reconstructed VF. Main Outcome Measures Structure–function relationship. Results The corrected Akaike information criterion values with threshold were found to be smaller than the threshold reconstructed with the VAE (thresholdVAE) in 9 of 10 sectors. A significant relationship was found between threshold and cpRNFL thickness in 6 of 10 sectors, whereas it was significant in 9 of 10 sectors with thresholdVAE. Conclusions Applying VAE to VF data results in an improved structure–function relationship.

The purpose of the current study was to investigate the structure-function relationship in patients with retinitis pigmentosa (RP) using optical coherence tomography and the MP-3 microperimeter. Visual field (VF) measurements were carried out using MP-3 microperimetry and the Humphrey Field Analyzer (HFA, Carl-Zeiss, CA), 22 eyes of 11 patients with a clinical diagnosis of RP, both with the 10-2 test grid pattern. Optical coherence tomography (OCT, Spectralis, Heidelberg, Germany) was also performed and the ellipsoid zone (EZ) was identified in the OCT image. The mean (±SD) number of test points located within the EZ edge was 11.6 (±5.9). There was a significant relationship between mean retinal sensitivity measured with MP-3 and the area surrounded by the EZ circular line: AEZ (p 0.05). The difference between retinal sensitivity inside and outside the EZ edge was significantly larger with MP-3 than with HFA (p

Erlotinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), shows superior efficacy in patients with non-small cell lung cancer (NSCLC) harboring activating EGFR mutations (EGFR Mut+). However, almost all tumors eventually develop resistance to erlotinib. Recently, the Phase II JO25567 study reported significant prolongation of progression-free survival (PFS) by erlotinib plus bevacizumab combination compared with erlotinib in EGFR Mut+ NSCLC. Herein, we established a preclinical model which became refractory to erlotinib after long-term administration and elucidated the mode of action of this combination. In this model, tumor regrowth occurred after remarkable shrinkage by erlotinib; regrowth was successfully inhibited by erlotinib plus bevacizumab. Tumor vascular endothelial growth factor (VEGF) was greatly reduced by erlotinib in the erlotinib-sensitive phase but significantly increased in the erlotinib-refractory phase despite continued treatment with erlotinib. Although EGFR phosphorylation remained suppressed in the erlotinib-refractory phase, phosphorylated extracellular signal-regulated kinase (pERK), phosphorylated AKT, and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) were markedly higher than in the erlotinib-sensitive phase; among these, pERK was suppressed by erlotinib plus bevacizumab. MVD was decreased significantly more with erlotinib plus bevacizumab than with each drug alone. In conclusion, the erlotinib plus bevacizumab combination demonstrated promising efficacy in the B901L xenograft model of EGFR Mut+ NSCLC. Re-induction of VEGF and subsequent direct or indirect VEGF-dependent tumor growth was suggested as a major mechanism of erlotinib resistance, and erlotinib plus bevacizumab achieved remarkably prolonged antitumor activity in this model.

Background Bevacizumab in combination with chemotherapeutics has shown significant survival benefit in clinical studies in patients with non-small cell lung cancer (NSCLC). Since bevacizumab was administered as standard treatment until disease progression, the importance of bevacizumab during the maintenance phase was not prospectively investigated in these studies. Materials and methods Three human NSCLC cell line xenograft models were used to investigate antitumor effect of bevacizumab and tumor microvessel density (MVD) was analyzed. Results In A549 and NCI-H292 models, bevacizumab maintenance following combination with paclitaxel exhibited stronger tumor suppression than its absence. In an NCI-H292 model, bevacizumab maintenance continuously inhibited increase of MVD. In an NCI-H2228 model following induction treatment with pemetrexed and bevacizumab, maintenance with pemetrexed plus bevacizumab, had stronger efficacy than pemetrexed alone and led to lower MVD and level of thymidylate synthase. Conclusion Continuous suppression of angiogenesis by bevacizumab may contribute to the superior efficacy of maintenance treatment containing bevacizumab.

Trastuzumab, a humanized anti-human epidermal growth factor receptor 2 antibody drug, is the first-line therapy for human epidermal growth factor receptor 2-positive breast and gastric cancer. For breast cancer, the benefit of continuous treatment with trastuzumab after it becomes refractory to first-line therapy has been demonstrated. However, it is unclear whether trastuzumab can show similar efficacy as a second-line treatment for gastric cancer. Here, we report that trastuzumab in combination with paclitaxel exhibits increased antitumor efficacy even for trastuzumab-resistant xenografted tumors. We derived the trastuzumab-resistant models from previously established human epidermal growth factor receptor 2-positive gastric cancer patient-derived cells. Human epidermal growth factor receptor 2 expression, PIK3CA mutation, and phosphatase and tensin homolog expression in these resistant models was equivalent to those in the trastuzumab-sensitive parental model, whereas cyclin-dependent kinase inhibitors, such as p16, p15, and p21, were downregulated. Trastuzumab in combination with paclitaxel enhanced antitumor activity in both the sensitive and resistant models. In the trastuzumab-sensitive model, the combination of trastuzumab and paclitaxel resulted in suppression of the AKT-p27-retinoblastoma protein pathway and induction of apoptosis. Although this combination did not suppress retinoblastoma protein phosphorylation in the trastuzumab-resistant model, it did markedly decrease epidermal growth factor receptor and human epidermal growth factor receptor 2 phosphorylation and further enhance paclitaxel-mediated apoptosis. These results suggested that trastuzumab in combination with paclitaxel can still exert more potent antitumor efficacy than each agent alone in trastuzumab-resistant models, providing evidence that trastuzumab remains beneficial in the treatment of trastuzumab-resistant tumors.

Although bevacizumab maintenance following bevacizumab in combination with chemotherapy has demonstrated significant prolongation of progression-free survival in clinical studies in patients with ovarian cancer, the majority of the cancer cases in the study were of the serous histotype; therefore, data regarding clear cell carcinoma is limited. Furthermore, the efficacy of bevacizumab beyond progression has not yet been demonstrated in ovarian cancer. A xenograft model using the human ovarian clear cell carcinoma cell line RMG-I was used to investigate the antitumor effects and the mechanisms of bevacizumab in maintenance treatment and bevacizumab when administered beyond disease progression. In the RMG-I model, bevacizumab maintenance following bevacizumab in combination with paclitaxel exhibited increased tumor suppression, compared with its absence, and inhibited the increase of microvessel density (MVD) in tumors. Following disease progression during bevacizumab maintenance, continued bevacizumab treatment in combination with PEGylated liposomal doxorubicin as a secondary chemotherapeutic agent had increased efficacy, compared with PEGylated liposomal doxorubicin alone, and resulted in lower MVD accompanied with lower levels of insulin-like growth factor binding protein-3, which is reported to have angiogenic activity. Continuous suppression of angiogenesis by bevacizumab may contribute to the superior efficacy of bevacizumab maintenance and bevacizumab beyond progression in ovarian cancer.

AimsCorvis ST (CST) yields biomechanical corrected IOP (bIOP) which is purported to be less dependent on biomechanical properties. In our accompanied paper, it was suggested that the repeatability of bIOP is high. The purpose of the current study was to assess the relationship between intraocular pressure (IOP) measured with CST and central corneal thickness (CCT) and corneal hysteresis (CH), in comparison with IOP measured with Goldmann applanation tonometry (GAT) and the ocular response analyzer (ORA).MethodsA total of 141 eyes from 141 subjects (35 healthy eyes and 106 glaucomatous eyes) underwent IOP measurements with GAT, CST and ORA. The relationships between IOP measurements (ORA-IOPg, ORA-IOPcc, CST-bIOP and GAT IOP) and biomechanical properties (CCT, CH and corneal resistance factor (CRF)) were analysed using the linear regression analysis.ResultsIOPg, IOPcc and GAT IOP were significantly associated with CCT (pConclusionThe bIOP measurement from CST is independent from CCT, but dependent on CH and CRF.

Purpose To investigate if the structure-function relationship between circumpapillary retinal nerve fibre layer (cpRNFL) thickness and visual field (VF) thresholds is stronger when using the Goldmann V target rather than the Goldman III target where glaucomatous damage is advanced. Methods Optical coherence tomography (OCT) and VF (Humphrey Field Analyzer 24-2 or 30-2) measurements with Goldmann III (SITA standard) and V (full-threshold) targets were carried out in 51 eyes of 51 patients with primary open angle glaucoma. The relationship between cpRNFL thicknesses in supero- and infero-temporal sectors, and VF sensitivity with the Goldmann III or V target was investigated. Result Visual field sensitivities (dB) both with the Goldmann III target and Goldmann V target showed a floor effect in the structure-function relationship against cpRNFL thickness, at approximately 60 μm. There was no significant relationship between visual field sensitivity measured with the Goldmann V target (dB scale: p = 0.12, 1/Lambert scale: p = 0.40; linear mixed models) and cpRNFL thickness, when corresponding visual field sensitivity, measured with the Goldmann III target, was Conclusion There was no improvement in the structure-function relationship using the Goldmann V target (full-threshold), compared to using the Goldmann III target (SITA standard), where glaucomatous damage was advanced.

Purpose To compare structure-function relationships based on the Drasdo and Sjöstrand retinal ganglion cell displacement models. Methods Single eyes from 305 patients with glaucoma and 55 heathy participants were included in this multicenter, cross-sectional study. The ganglion cell and inner plexiform layer (GCIPL) thickness was measured using spectral domain optical coherence tomography. Visual field measurements were performed using the Humphrey 10-2 test. All A-scan pixels (128 × 512 pixels) were allocated to the closest 10-2 location with both displacement models using degree and millimeter scales. Structure-function relationships were investigated between GCIPL thickness and corresponding visual sensitivity in nonlong (160 eyes) and long (200 eyes) axial length (AL) groups. Results In both the nonlong and long AL groups, compared with the no-displacement model, both the Drasdo and the Sjöstrand models showed that the structure-function relationship around the fovea improved (P < 0.05). The magnitude of improvement in the area was either comparable between the model or was larger for the Drasdo model than the Sjöstrand model (P < 0.05). Meanwhile, structure-function relationships outside the innermost retinal region that were based on the Drasdo and Sjöstrand models were comparable to or were even worse than (in the case of the Drasdo model) those obtained using the no-displacement model. Conclusions Structure-function relationships evaluated based on both the Drasdo and Sjöstrand models significantly improved around the fovea, particularly when using the Drasdo model. This was not the case in other areas.

This study investigated the relationship between the shape of the posterior pole of the eye and the vertical asymmetry of visual field (VF) damage in glaucomatous eyes.Cross-sectional study.A total of 83 eyes of 43 patients with open-angle glaucoma.The VF was measured using the Humphrey 24-2 or 30-2 Swedish Interactive Thresholding Algorithm (SITA) standard. The vertical asymmetry of VF damage was defined as the difference between the mean total deviation (mTD) values of the superior and inferior hemi-retinas. This difference was calculated for the hemifield, central, middle, and peripheral areas. A vertical cross-sectional image of the macula was obtained, and the magnitudes of superior and inferior retinal tilt (RT) were calculated as the difference between the lower margins of the retinal pigment epithelium (RPE) at the fovea and at a location 2 mm superior or inferior to the RPE. The RT asymmetry was defined as the difference between the inferior and superior RT. Data were analyzed using a multivariate linear mixed model.The relationship between mTD asymmetry (hemifield, central, middle, and peripheral) and both RT asymmetry and other factors (intraocular pressure, axial length, corneal radius, age, gender, mean deviation, refraction, and visual acuity) was assessed.The inferior RT was significantly larger than the superior RT (P0.001). Hemifield mTD asymmetry was related to RT asymmetry (P = 0.017). These relationships were significant in the middle (P = 0.029) and peripheral areas (P = 0.023), but not in the central area (P = 0.40). Other factors were not related to mTD asymmetry.Vertical asymmetry of the posterior pole was related to the vertical asymmetry of glaucomatous VF damage.

Purpose To assess the repeatability of intraocular pressure (IOP) measured with the Corvis ST (CST) and the Ocular Response Analyzer (ORA). Methods A total of 141 eyes from 141 subjects were studied, including 35 healthy eyes and 106 glaucomatous eyes. All subjects underwent IOP evaluations with Goldmann applanation tonometer, CST, and ORA. With CST, biomechanical corrected IOP (bIOP) was calculated; bIOP is purported to be less dependent on biomechanical properties. For ORA, corneal-compensated intraocular pressure (IOPcc) and Goldmann-correlated IOP (IOPg) were derived. The repeatability of the various IOP values was assessed using the coefficient of variance (CV) and the intraclass correlation coefficient (ICC). Results The CV with bIOP (5.5 ± 3.1: mean ± standard deviation) was significantly smaller than the CVs measured with IOPg (7.3 ± 4.3) and IOPcc (7.2 ± 4.4). ICC values were 0.90, 0.80, and 0.86 with IOPg, IOPcc, and bIOP, respectively. Conclusions The bIOP showed a better prevision and repeatability for IOP measurement. Translational Relevance The bIOP measurement from CST had a better reproducible than IOPcc measurement from ORA.

The purpose of the study was to investigate whether the structure-function relationship in glaucomatous and normative eyes is improved by considering photoreceptor layer thickness. Humphrey 10-2 visual fields (VF) and optical coherence tomography were carried out in 615 eyes of 391 subjects, including 100 eyes of 53 healthy controls and 515 eyes of 338 glaucoma patients. The relationship between mean VF sensitivity and the thickness of the retinal nerve fiber layer (RNFL) and ganglion cell layer and inner plexiform layer (GCL + IPL) was analyzed using linear mixed models, by glaucoma status and degree of myopia. The structure-function relationship was also analyzed by supplementing the RNFL and GCL + IPL thickness with the thicknesses of: (i) the inner nuclear layer and outer plexiform layer (INL + OPL); (ii) the outer nuclear layer and inner segment of photoreceptor layer (ONL + ISL); (iii) the outer segment layer of photoreceptor and retinal pigment epithelium (OSL + RPE). The model included total thickness of RNFL, GCL + IPL and OSL + RPE was highly more optimal than the model that only included the total thickness of RNFL and GCL + IPL, in all subsets of eyes by glaucoma status and degree of myopia.

Erlotinib (ERL), an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, shows notable efficacy against non-small cell lung cancer (NSCLC) harboring EGFR mutations. Bevacizumab (BEV), a humanized monoclonal antibody to vascular endothelial cell growth factor (VEGF), in combination with ERL (BEV+ERL) significantly extended progression-free survival in patients with EGFR-mutated NSCLC compared with ERL alone. However, the efficacy of BEV+ERL against EGFR-mutated NSCLC harboring T790M mutation or MET amplification, is unclear. Here, we examined the antitumor activity of BEV+ERL in four xenograft models of EGFR-mutated NSCLC (three harboring ERL resistance mutations). In the HCC827 models (exon 19 deletion: DEL), ERL significantly inhibited tumor growth by blocking EGFR signal transduction. Although there was no difference between ERL and BEV+ERL in maximum tumor growth inhibition, BEV+ERL significantly suppressed tumor regrowth during a drug-cessation period. In the HCC827-EPR model (DEL+T790M) and HCC827-vTR model (DEL+MET amplification), ERL reduced EGFR signal transduction and showed less pronounced but still significant tumor growth inhibition than in the HCC827 model. In these models, tumor growth inhibition was significantly stronger with BEV+ERL than with each single agent. In the NCI-H1975 model (L858R+T790M), ERL did not inhibit growth or EGFR signal transduction, and BEV+ERL did not inhibit growth more than BEV. BEV alone significantly decreased microvessel density in each tumor. In conclusion, addition of BEV to ERL did not enhance antitumor activity in primarily ERL-resistant tumors with T790M mutation; however, BEV+ERL enhanced antitumor activity in T790M mutation- or MET amplification-positive tumors as long as their growth remained significantly suppressed by ERL.

In the treatment of human epidermal growth factor receptor 2 (HER2)-positive advanced gastric or gastroesophageal junction cancer, it has been reported that the combination of trastuzumab with capecitabine plus cisplatin, or with 5-fluorouracil (5-FU) plus cisplatin, significantly increased overall survival compared with chemotherapy alone (ToGA trial). In addition, adjuvant therapy with capecitabine plus oxaliplatin (XELOX) improved the survival of patients who received curative D2 gastrectomy (CLASSIC trial). However, the efficacy of the combination of trastuzumab with XELOX for patients with HER2-positive gastric cancer remains unknown. The aim of this study, was to investigate the efficacy of the combination of trastuzumab with XELOX in a HER2-positive human gastric cancer xenograft model. Combination treatment with these three agents (trastuzumab 20 mg/kg, capecitabine 359 mg/kg and oxaliplatin 10 mg/kg), was found to exhibit a significantly stronger antitumor activity in NCI-N87 xenografts compared with either trastuzumab or XELOX alone. In this model, treatment with trastuzumab alone or trastuzumab plus oxaliplatin enhanced the expression of thymidine phosphorylase (TP), a key enzyme in the generation of 5-FU from capecitabine in tumor tissues. In in vitro experiments, trastuzumab induced TP mRNA expression in NCI-N87 cells. In addition, NCI-N87 cells co-cultured with the natural killer (NK) cell line CD16(158V)/NK-92 exhibited increased expression of TP mRNA. When NCI-N87 cells were cultured with CD16(158V)/NK-92 cells in the presence of trastuzumab, the mRNA expression of cytokines reported to have the ability to induce TP was upregulated in tumor cells. Furthermore, a medium conditioned by CD16(158V)/NK-92 cells also upregulated the expression of TP mRNA in NCI-N87 cells. These results suggest that trastuzumab promotes TP expression, either by acting directly on NCI-N87 cells, or indirectly via a mechanism that includes trastuzumab-mediated interactions between NK and NCI-N87 cells. Therefore, the combination of trastuzumab with XELOX may be a potent therapy for HER2-positive gastric cancer.

Purpose To investigate the relationship between axial length (AL) elongation and progression of primary open-angle glaucoma (POAG). Methods AL was measured twice over a 5.1 ± 0.76 (mean ± standard deviation: SD) year period in 125 eyes of 72 patients with POAG. The eyes were divided into not long (AL 26 mm, 45 eyes) groups. During this period, patients' visual fields (VFs) were measured with the Humphrey Field Analyzer 12.4 ± 7.5 times and intraocular pressure (IOP) was measured with Goldmann applanation tonometry 27.0 ± 7.5 times. The relationship between the mean total deviation (mTD) progression rates in the whole field and superior and inferior hemifield, as well as in 10 VF sectors, and the variables of age, mean IOP, SD of IOP, AL, difference in AL (△AL), and mTD value at baseline was examined. Results There was a significant difference between AL at baseline and AL at repeat measurement (P < 0.0001). △AL was 0.035 ± 0.10 mm. An increase in △AL was significantly related to AL at baseline (P = 0.027), but not to age, mean IOP, and SD of IOP. △AL was related to the progression of mTD in the inferior hemifield (slower mTD progression was associated with increased △AL), but not in the whole field or superior hemifield. Increased △AL was related to slower progression rates in 2 of 10 sectors, both in the inferior hemifield. Conclusions The main finding was that an increase in AL was significantly related to slower VF progression in the inferior hemifield.

PURPOSE To compare the Esterman Disability Score (EDS) obtained with Goldmann perimetry (GP) testing and the Humphrey field analyzer (HFA) in low vision Japanese subjects. Subjects were also divided into groups by diagnosis to examine how disease influences EDS measurements. METHODS The EDS was obtained using GP (GP-EDS) and the built-in testing program of the HFA (HFA-EDS). Tests were performed within 3 months of each other. Regression analyses were used to examine the relationship between GP-EDS and HFA-EDS. RESULTS A total of 128 visually impaired subjects were included in this study. Subjects had low vision because of glaucoma (57 subjects), age-related macular degeneration (AMD, 17 subjects), retinitis pigmentosa (RP, 17 subjects), and other causes (37 subjects). The GP-EDS obtained was well-correlated with HFA-EDS (r = 0.87, P < 0.001) and it was possible to estimate HFA-EDS from GP-EDS. The GP-EDS was significantly lower than the HFA-EDS in eyes with glaucoma and RP. There was no significant difference between EDS values in eyes with AMD or other disease. CONCLUSION The GP-EDS correlated well with the HFA-EDS. However, the relationship between the EDS measured with the two different testing modalities varies by disease.

Purpose To investigate the relationship between visual field (VF) loss and the rings of the ellipsoid zone (EZ) and autofluorescence (AF) in patients with retinitis pigmentosa (RP). Design Cross-sectional study. Participants A total of 28 eyes of 15 patients with a clinical diagnosis of RP. Methods The VF was measured at a 1-degree interval on the horizontal or vertical lines, within the central 10 degrees from fixation, using MP-3 microperimetry (Nidek Co. Ltd., Aichi, Japan), and the sensitivity deviation from the normative database was calculated. OCT was performed, and the EZ was identified. Fundus AF was also measured, and the AF ring was identified. The VF test points were categorized according to their relationship with the EZ line and AF ring. The relationship between mean retinal sensitivity deviations on both the horizontal and vertical lines and the diameters of the EZ line and the AF ring in the corresponding directions was analyzed. The relationship between visual sensitivity deviation and the locations against EZ line and AF rings was also investigated. Main Outcome Measures The relationship between the visual sensitivity deviation and the locations against EZ line and AF rings. Results The diameter of the AF ring had a significantly stronger relationship with mean retinal sensitivity deviation compared with the diameter of the EZ line in both the horizontal and vertical directions. Test point location inside/on/outside the AF ring was significantly related to retinal sensitivity deviation independent of the location inside/outside the EZ line (P Conclusions Visual sensitivity damage is significantly associated with the EZ lines and AF rings. The development of the AF ring is related to the decrease of visual function independent of and stronger than the EZ line.

The purpose of the current study was to evaluate the test-retest reproducibility and structure-function relationship of the MP-3 microperimeter, compared against the Humphrey Field Analyzer (HFA).Design: Reliability and validity study.Institutional, or clinical practice.Thirty eyes of 30 primary open-angle glaucoma patients were enrolled.Visual fields (VF) were measured twice with the MP-3 and HFA instruments, using the 10-2 test grid pattern in both perimeters. Ganglion cell complex (GCC) thickness was measured using optical coherence tomography (OCT). Test-retest reproducibility was assessed using the mean absolute deviation (MAD) measure at all 68 VF test points, and also the intraclass correlation coefficient (ICC) of the repeated VF sensitivities. The structure-function relationship between VF sensitivities (measured with MP-3 or HFA) and GCC thickness (adjusted for the retinal ganglion cell displacement) was analyzed using linear mixed modeling.Reproducibility and structure-function relationship.The average measurement duration with the HFA 10-2 was 7 minutes and 6 seconds (7m06s) ± 0m49s (mean ± standard deviation). A significantly (P .001, paired Wilcoxon test) longer measurement duration was observed for the MP-3 test: 10m29s ± 2m55s. There were no significant differences in MAD and ICC values between HFA (MAD; 0.83 ± 0.69 dB and ICC: 0.89 ± 0.69, mean ± standard deviation) and MP-3 (MAD: 0.65 ± 0.67 dB and ICC: 0.89 ± 0.69). MP-3 VF sensitivities had a stronger structure-function relationship with GCC thickness compared to HFA.The MP-3 microperimeter has a similar test-retest reproducibility to the HFA but a better structure-function relationship.

PURPOSE To investigate the effect of axial length (AL) on the structure-function relationship between retinal nerve fiber layer (RNFL) thickness measurements and visual field (VF) sensitivity measured with Goldmann III and V. METHOD There were 85 eyes of 85 patients with primary open angle glaucoma included in the current study. Optical coherence tomography and VF (Humphrey Field Analyzer 24-2 or 30-2) measurements with Goldmann III and V targets were carried out in all patients. The optic disc and the VF were divided into six clusters and the relationship between circumpapillary RNFL (cpRNFL) thickness and VF sensitivity (with Goldmann III or V), age, and AL were investigated in each cluster. RESULT Visual sensitivity with Goldmann III (19.3 ± 11.7 dB, mean ± standard deviation) was significantly lower than that with Goldmann V (24.6 ± 11.0 dB, P < 0.001, linear mixed model). Visual sensitivities with both Goldmann III and V were significantly correlated with cpRNFL thickness in all clusters. Visual sensitivity decreased with increasing AL in the nasal retinal area for both targets, however, this phenomenon was only observed with the Goldmann V target in the temporal area. CONCLUSION Visual sensitivity measured with the size V target decreases with increasing AL in the temporal area, which corresponds to the papillomacular bundle. In the nasal retinal area, visual sensitivity decreases with the increase of AL for both Goldmann III and Goldmann V. TRANSLATIONAL RELEVANCE Careful consideration is needed when measuring visual sensitivity using Goldmann V target in glaucomatous eyes with increased AL.

We examined the antitumor efficacy of the capecitabine (CAPE) plus cyclophosphamide (CPA) combination as a 2nd-line therapy after paclitaxel (PTX) plus bevacizumab (BEV) treatment in a xenograft model of human triple negative breast cancer (TNBC) cell line, MX-1. After tumor growth was confirmed, PTX (20 mg/kg; i.v.) + BEV (5 mg/kg; i.p.) treatment was started (Day 1). Each agent was administered once a week for 5 weeks and tumor regression was observed for at least the first 3 weeks. For 2nd-line treatment, we selected mice in which the tumor volume had increased from day 29 to day 36 and was within 130 - 250 mm3 on day 36. After randomization of mice selected on day 36, CPA (10 mg/kg; p.o.) and CAPE (539 mg/kg; p.o.) were administered daily for 14 days (days 36 - 49), followed by cessation of the drugs for 1 week. The tumor growth on day 57 was significantly suppressed in the CPA, CAPE and CAPE + CPA groups as compared with the control group (p < 0.05). Furthermore, the antitumor activity on day 57 of CAPE + CPA was significantly stronger than that of CPA or CAPE alone (p < 0.05). The thymidine phosphorylase (TP) level in tumor tissue was evaluated by immunohistochemistry on day 50, and was significantly higher in the CPA group than those in the control group (p < 0.05). Upregulation of TP in tumor tissues by CPA treatment would increase the 5-FU level in tumor tissues treated with CAPE. This would explain the possible mechanism that made CAPE + CPA superior to CAPE alone in the 2nd-line treatment. Our preclinical results suggest that the CAPE + CPA combination therapy may be effective as 2nd-line therapy after disease progression in PTX + BEV 1st-line treatment for TNBC patients.

PURPOSE To investigate whether it is possible to improve estimation of the binocular visual field (VF) using monocular sensitivities on a linear scale adjusted for ocular dominance. METHODS Monocular and binocular VF measurements were evaluated using the Humphrey Field Analyzer (HFA; 24-2 Swedish Interactive Threshold Algorithm standard program) in 60 eyes of 30 patients with open angle glaucoma. Ocular dominance was measured twice in each patient and the average value was used. Measured binocular sensitivity was then predicted based on monocular measurements using the "better sensitivity" integrated visual field (IVF) method, monocular sensitivity summation methods on the dB scale, linear scale (1/Lambert), and finally monocular sensitivity summation methods on the linear scale adjusted for the ocular dominance. RESULTS The absolute prediction error with the linear scale summation method (mean ± SD: 3.11 ± 4.00) was significantly smaller than the IVF method (3.15 ± 4.09; P = 0.014). Further, the absolute prediction error for the ocular dominance adjusted method (3.10 ± 3.99) was significantly smaller than the nonadjusted linear scale summation method (P = 0.014). The absolute prediction error associated with the dB scale summation method was significantly larger than any other method (8.15 ± 5.06; P < 0.0001). CONCLUSIONS The most accurate estimation of binocular sensitivity was achieved using the linear monocular sensitivity summation model adjusted for ocular dominance.

Molecular markers predicting sensitivity to anticancer drugs are important and useful not only for selecting potential responders but also for developing new combinations. In the present study, we analyzed the difference in the sensitivity of xenograft models to capecitabine (Xeloda®), 5'-deoxy-5-fluorouridine (5'-DFUR, doxifluridine, Furtulon®) and 5-FU by comparing the mRNA levels of 12 pyrimidine nucleoside-metabolizing enzymes. Amounts of mRNA in the tumor tissues of 80 xenograft models were determined by real-time RT-PCR and mutual correlations were examined. A clustering analysis revealed that the 12 enzymes were divided into two groups; one group consisted of 8 enzymes, including orotate phosphoribosyl transferase (OPRT), TMP kinase (TMPK) and UMP kinase (UMPK), and was related to the de novo synthesis pathway for nucleotides, with mRNA expression levels showing significant mutual correlation. In the other group, 4 enzymes, including thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD), were involved in the salvage/degradation pathway of the nucleotides, and the mRNA levels of this group were dispersed more widely than that of the de novo group. Antitumor activity was assessed in 24 xenograft models for each drug. The antitumor activity of capecitabine and 5'-DFUR correlated significantly with the mRNA levels of TP and with the TP/DPD ratio, whereas the activity of 5-FU correlated significantly with OPRT, TMPK, UMPK and CD. In a stepwise regression analysis, TP and DPD were found to be independent predictive factors of sensitivity to capecitabine and 5'-DFUR, and UMPK was predictive of sensitivity to 5-FU. These results indicate that the predictive factors for sensitivity to capecitabine and 5'-DFUR in xenograft models may be different from those for 5-FU, suggesting that these drugs may have different responders in clinical usage.

To investigate the vision-related quality of life (VRQOL) in relation to the type of existing visual field (EVF) in the presence of visual field defect. We evaluated the VRQOL of 95 patients by using Visual Function Questionnaire-25 (NEI VFQ-25). Patients fulfilled the following criteria: (1) they underwent a Goldmann perimetry (GP) test within 3 months of the initial visit, (2) the results of the GP test could be classified into three categories, and (3) the best-corrected decimal visual acuity (VA) was ≤0.3. The EVF was assessed by composite measurement of both eyes for the V-4e or I-4e isopters, as obtained using the GP test. We compared the VFQ-25 scores between three groups: (1) the "center and peripheral" group in which the EVF of the V-4e area was30º and the I-4e area was30º, (2) the "central VF" group in which the EVF of the V-4e area was20º, and (3) the "central VF loss" group in which the central scotoma was5º and the EVF of the V-4e area was30º in peripheral vision. There were significant differences between the three groups (P = 0.02, ANOVA). The total score of the "central VF loss" group was significantly lower than those of the "center and peripheral" and "central VF" groups. The central vision influenced the VRQOL to a greater extent than peripheral vision.

Purpose: To investigate the association between vision-related quality of life (VRQOL) and changes in visual acuity (VA). Methods: We examined the VA in 100 patients for >1 year and evaluated the degree of its impact on VRQOL using the National Eye Institute Visual Function (VF) Questionnaire (VFQ-25; Japanese version). Before determining VFQ-25, we monitored the changes in VA in these patients for 1 year and classified them into the following two groups depending on VA changes. Patients exhibiting a decline of >3 steps in VA, as assessed by the logarithm of the minimum angle of resolution scale, were placed in the ‘decline’ group (47.0%) and patients exhibiting no change in VA were placed in the ‘no change’ group (53.0%). We compared the VFQ-25 scores between both groups in all patients with glaucoma (GLA) and macular degeneration (MD). Results: The total score of the decline was 34.9 ± 13.6 and that of the no change group was 44.6 ± 13.9: the difference in the scores between both groups was statistically significant (p = 0.006). Similar results were obtained for patients with GLA and MD (p = 0.007 and 0.003, respectively). Conclusion: VRQOL differed between patients with constant VA and those with reduced VA, even though VA values were equal at a certain time point.

It has been reported that bevacizumab in combination with paclitaxel significantly prolongs progression-free survival compared with paclitaxel alone in the initial treatment for metastatic breast cancer. To understand how bevacizumab enhances the efficacy of paclitaxel, we investigated the mechanism in a MX-1 human breast cancer xenograft model. The antitumor activity of bevacizumab at 5 mg/kg in combination with paclitaxel at 20 or 30 mg/kg was significantly higher than that of either agent alone. First, we measured the paclitaxel concentration in tumor to see whether bevacizumab enhances the activity by increasing the tumor concentration of paclitaxel. When given in combination with bevacizumab, the levels of paclitaxel in the tumor increased. Paclitaxel at 30 mg/kg with bevacizumab showed a similar tumor concentration as paclitaxel alone at either 60 or 100 mg/kg, with a similar degree of tumor growth inhibition. In contrast, no remarkable differences in paclitaxel concentration in the plasma or liver were observed between the paclitaxel monotherapy group and the paclitaxel plus bevacizumab group. An increase in paclitaxel concentration by bevacizumab was also found in another model, A549. In the same MX-1 model, vascular permeability in the tumor was significantly decreased by treatment with bevacizumab. There was no difference in microvessel density between the bevacizumab alone group and the combination group. Results suggest that the synergistic antitumor activity of paclitaxel and bevacizumab in combination may be a result of the increase in paclitaxel concentration in tumor resulting from the downregulation of vascular permeability when co-administered with bevacizumab.

Purpose To investigate the structure-function mapping in the central 10° by relating Humphrey field analyzer (HFA) 10-2 visual field (VF) and circumpapillary retinal nerve fiber layer (cpRNFL) thickness from spectral-domain optical coherence tomography (SD-OCT). We also compared the obtained results with a previously reported mapping between 10-2 VF and the optic disc. Methods In 151 eyes of 151 POAG patients and 35 eyes from 35 healthy participants, cpRNFL thickness measurements were obtained using SD-OCT and the 10-2 VF was measured with the HFA. The relationship between visual sensitivity and cpRNL thickness values in the temporal 180° was analyzed using least absolute shrinkage and selection operator (LASSO) regression. The optic disc angle corresponding to each VF test point was then derived using the coefficients from the optimal LASSO regression. Results The structure-function map obtained was largely consistent with the mapping reported previously; superior central VF test points correspond to a more vulnerable area of the optic disc, more distant toward the inferior pole from the center of the temporal quadrant (9:00 o'clock for the right eye) while inferior VF test points correspond closer to the center of the temporal quadrant. The prediction error tended to be large in the 'more vulnerable area' in the map reported previously. Conclusions The structure-function map obtained largely confirms the previously reported map; however, some important differences were observed.

256 Background: Trastuzumab emtansine (T-DM1) is an antibody-drug conjugate composed of trastuzumab and DM1 (an anti-microtubule agent derived from maytansine) that is approved for the treatment of HER2-positive metastatic breast cancer. HER2-overexpression rate in biliary tract cancer (BTC) is reported as approximately 10%, and HER2-targeted therapy may enhance therapeutic efficacy in HER2-positive BTC. In this study, we investigated the antitumor activity of T-DM1 in several BTC cell lines and xenograft models. Methods: HER2 expression was detected by flow cytometry and Western blotting in 17 BTC cell lines. The cell growth inhibition activity of T-DM1 and DM1 was examined by quantifying the DNA with Hoechst 33258 nucleic acid stain. In vivo antitumor activity of T-DM1 was evaluated in three xenograft mouse models using cell lines with different levels of HER2 expression. HER2 expression in xenografts was assessed by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), according to gastric cancer criteria. Cell cycle was assessed by flow cytometry after staining with propidium iodide. Apoptosis was assessed by caspase 3/7 activity measurement. Results: In the 17 BTC cell lines, HER2 expression was high in KMCH-1, Mz-ChA-1, and TGBC-18-TKB. Although cell growth inhibition of DM1 was similar in all cell lines (50% inhibitory concentration (IC50): 0.79–7.2 nM), that of T-DM1 was HER2-expression-dependent, with IC50 values in KMCH-1, Mz-ChA-1, and KKU-100 (which has low HER2 expression) of 0.031, 1.3, and 4.3 μg/mL, respectively. In KMCH-1 and Mz-ChA-1 xenograft models, T-DM1 treatment once every 3 weeks for 6 weeks showed significant dose-dependent antitumor activity, and tumor growth inhibition 21 days after first dose at 20 mg/kg T-DM1 was 108% (KMCH-1) and 75% (Mz-ChA-1), whereas there was no significant efficacy in KKU-100 xenograft model. Values for HER2 status (IHC score/FISH ratio of HER2 to CEP17) in KMCH-1, Mz-ChA-1, and KKU-100 tumor tissues were 3+/3.5, 2+/4.7, and 0/1.1, respectively. In KMCH-1, T-DM1 induced M phase arrest and apoptosis, as is reported in HER2-positive breast cancer cells. Conclusions: T-DM1 could be an effective therapy for HER2-overexpressed BTC.

To evaluate the sensitivity and specificity with glaucoma probability score (GPS), a new glaucoma diagnosis program in Heidelberg Retina Tomograph II ver 3.0 in detecting early glaucoma in Japanese eyes.Sensitivity and specificity with GPS were determined in 148 early stage open-angle glaucoma eyes of 148 hospital-based patients (mean deviation-5 dB) and age-matched and refraction-matched 148 eyes of 148 ophthalmologically normal subjects selected from a population-based sample of the Tajimi Study. The results with GPS were compared with those with Moorfields regression analysis (MRA) and FS Mikelberg discriminant function (FSM). Analysis was also carried out by stratifying the eyes by refraction and disc area.For all glaucoma eyes, sensitivity with GPS (71.7%) was similar to MRA and FSM (75.2%, P=0.8; 77.2%, P=0.5; Fisher exact test). Specificity with GPS (80.7%) was also statistically equivalent with MRA and FSM (91.1%, P=0.06; 87.4%, P=0.3). Although sensitivity with GPS was lower in higher myopic eyes than in the less myopic eyes, specificity did not differ with refractive error. Specificity with GPS was lower and sensitivity higher (P0.0001) in larger discs (2.0 mm) than in smaller discs (or =2 mm). In larger discs, specificity with GPS was lower (P=0.005) than with MRA and in smaller discs, sensitivity with GPS was lower than with MRA or FSM.In Japanese eyes, sensitivity and specificity with GPS to detect early glaucoma were similar with MRA or FSM, although the specificity tended to be lower than results in white eyes. Subgroup analysis revealed a notable dependency in diagnostic ability on disc area and refractive error. The diagnostic ability of GPS is still not sufficient for glaucoma detection in the Japanese population requiring further investigation.

To observe the morphology of the corneal cells and corneal nerve fibers in patients with type IV or V hereditary sensory and autonomic neuropathy (HSAN) by in vivo confocal microscopy and elucidate the mechanism leading to the loss of corneal sensation in this disease.In vivo confocal microscopy was performed on the central cornea of the right eye in 3 patients with HSAN (ages 17, 20, and 32 years), and their corneal morphology was compared with that of 3 healthy subjects (ages 28, 30, and 36 years). Corneal sensation was tested with a Cochet-Bonnet esthesiometer.The superficial epithelial cell density was lower in the HSAN patients compared with the healthy subjects (1525, 1225, and 1250/mm(2) vs. 2225, 1750, and 2500/mm(2)), but the basal epithelial cell density of the patients was similar to that of the healthy subjects. Nerve bundles were clearly observed in the sub-basal nerve plexus layer of the cornea in the healthy subjects, but were undetectable at the central cornea in the patients with HSAN. The corneal sensation of the patients with HSAN was much weaker than that of the healthy subjects (2.79, 40.30, and 132.50 g/mm(2) vs. 1.47, 1.47, and 1.47 g/mm(2)).Superficial keratopathy accompanied with neurotrophic keratopathy and tear film instability observed clinically agrees with the large keratinized cells in the superficial corneal epithelium by in vivo confocal microscopy in these patients. Our findings suggest that the loss of corneal nerves contributes to impairment of corneal sensation in patients with type IV or V HSAN.

It is important to compare the results of Corneal Visualization Scheimpflug Technology instrument (CST) measurements and Reichert Ocular Response Analyzer (ORA) parameters. The purpose of the study was to investigate the association between CST measurements and ORA parameters in ninety-five patients with primary open-angle glaucoma. Measurements of CST, ORA, axial length (AL), average corneal curvature (CC), central corneal thickness (CCT) and intraocular pressure (IOP) with Goldmann applanation tonometry (GAT) were carried out. The association between CST and ORA parameters was assessed using linear regression analysis, with model selection based on the second order bias corrected Akaike Information Criterion index. Measurements from ORA (corneal hysteresis [CH] and corneal response factor [CRF]) had high intraclass correlation coefficients (ICC) and low coefficients of variation, but some CST parameters showed much lower reproducibility, namely: A1 length, A2 length, highest concavity time and peak distance. Of 12 CST parameters tested, 8 were significantly correlated with CH and 10 were significantly correlated with CRF, however, the magnitude of the correlation coefficients were weak to moderate at best. The optimal model to explain CH using CST measurements was given by: CH = -76.3 + 4.6*A1 time + 1.9*A2 time + 3.1 * highest concavity deformation amplitude + 0.016*CCT (R2 = 0.67, p

Erlotinib (ERL), an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, shows notable efficacy against non-small cell lung cancer (NSCLC) harboring EGFR mutations. Bevacizumab (BEV), a humanized monoclonal antibody to vascular endothelial cell growth factor (VEGF), in combination with ERL (BEV+ERL) significantly extended progression-free survival in patients with EGFR-mutated NSCLC compared with ERL alone. However, the efficacy of BEV+ERL against EGFR-mutated NSCLC harboring T790M mutation or MET amplification, is unclear. Here, we examined the antitumor activity of BEV+ERL in four xenograft models of EGFR-mutated NSCLC (three harboring ERL resistance mutations). In the HCC827 models (exon 19 deletion: DEL), ERL significantly inhibited tumor growth by blocking EGFR signal transduction. Although there was no difference between ERL and BEV+ERL in maximum tumor growth inhibition, BEV+ERL significantly suppressed tumor regrowth during a drug-cessation period. In the HCC827-EPR model (DEL+T790M) and HCC827-vTR model (DEL+MET amplification), ERL reduced EGFR signal transduction and showed less pronounced but still significant tumor growth inhibition than in the HCC827 model. In these models, tumor growth inhibition was significantly stronger with BEV+ERL than with each single agent. In the NCI-H1975 model (L858R+T790M), ERL did not inhibit growth or EGFR signal transduction, and BEV+ERL did not inhibit growth more than BEV. BEV alone significantly decreased microvessel density in each tumor. In conclusion, addition of BEV to ERL did not enhance antitumor activity in primarily ERL-resistant tumors with T790M mutation; however, BEV+ERL enhanced antitumor activity in T790M mutation- or MET amplification-positive tumors as long as their growth remained significantly suppressed by ERL.

Purpose: To examine the antitumor activity and tolerability of a combination comprising erlotinib and capecitabine in human colorectal, breast and epidermal cancer xenograft models. Further aims of the study were to examine the effects of single-agent erlotinib therapy on tumor growth, and on thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) levels, (enzymes which activate and deactivate capecitabine, respectively) in tumor tissue. Methods: BALB/c nu/nu mice bearing LoVo and HT-29 (colon cancer), A-431 (vulval cancer), and KPL-4 and MAXF401 (breast cancer) human tumors were treated with erlotinib 100 mg/kg/day and/or capecitabine 359 or 90 mg/kg/day, by oral administration once daily for 14 days. Results: The maximum tolerated dose (MTD) of erlotinib, formulated in carboxymethylcelluose/Tween 80, was identified as 125 mg/kg/day. Erlotinib at a dose of 100 mg/kg/day achieved significant tumor-growth inhibition in the, LoVo, KPL-4, and A-431 models. Some inhibition of MAXF401 tumor growth was observed, but was not significant. In the HT-29 model, erlotinib showed less marked but statistically significant antitumor activity. On day 15, mean tumor-growth inhibition in HT-29, LoVo, KPL-4, MAXF401, and A-431 models was 46, 74, 71, 20, and 85%, respectively. Evaluation of erlotinib/capecitabine combination therapy, at sub-optimal doses, in the three erlotinib-sensitive tumor models LoVo, KPL-4 and A-431, demonstrated at least additive activity with the combination compared with the single agents. In the A-431 and LoVo models, the combination of agents had greater antitumor activity than the single agent capecitabine alone at the MTD. Erlotinib in combination with capecitabine was not associated with significantly increased toxicity compared with single-agent therapy. Erlotinib 100 mg/kg/day induced significant upregulation of TP and DPD in the LoVo model, a significant upregulation of TP in the HT-29, MAXF401 and A-431 models, but had no obvious effect on TP and DPD levels in the KPL-4 model. In the A-431 model, selective upregulation of TP by erlotinib 100 mg/kg resulted in an increased TP:DPD ratio. In the LoVo model, immunohistochemistry revealed marked upregulation of TP (but not DPD by erlotinib). Conclusions: Erlotinib inhibits tumor growth in a range of human tumor xenograft models, including breast and colorectal cancer (CRC). Erlotinib and capecitabine demonstrated at least additive activity in LoVo, KPL-4 and A-431 tumor models. The antitumor activity of the combination was greater than that of capecitabine alone at the MTD. Erlotinib treatment did affect the TP in the CRC tumor models as confirmed immunohistochemically. The findings of this study support clinical evaluation of erlotinib, both as a single agent and in combination with capecitabine, for the treatment of CRC and breast cancer.

Our vision system receives two different images from our two eyes. Since these two images usually have high correlation they are fused into a single stable image. In binocular rivalry suppression a stimulus presented in one eye is suppressed and a different stimulus in the other eye is perceived for a certain time interval. For strabismic observers the suppression is continuous between different retinal images caused by misalignment of eye positions. Previous research investigated the effects of this binocular rivalry suppression on a number of visual processes. The present study looked at the effects of contrast adaptation for the adaptation stimulus presented to eyes under interocular suppression. We compared normal and strabismic observers and found that in both types, regardless of the existence of suppression, there was the effect of contrast adaptation. These findings provide evidence that the suppression occurs at a site in the visual system after the locus of contrast adaptation. The contrast adaptation strength differed in the two types of observers.

Purpose To assess the thickness of the photoreceptor layer in the macular region in glaucomatous eyes. Method Humphrey 10–2 visual field (VF) testing was carried out and mean threshold (mTH) was calculated in 118 eyes from 118 patients with open angle glaucoma. Macular optical coherence tomography (OCT) measurements (RS 3000, Nidek Co.ltd., Aichi, Japan) were also carried out in all eyes. Thickness measurements were recorded in the outer segment and retinal pigment epithelium (OS+RPE), the nerve fiber layer (NFL), the ganglion cell layer and inner plexiform layer (GCL+IPL), the inner nuclear layer and outer plexiform layer (INL+OPL) and the outer nuclear layer and inner segment (ONL+IS). The relationship between mTH and the thickness of these five different layers was investigated. Additionally, the influence of OS+RPE on mTH was investigated using partial correlation eliminating the effect of other variables of NFL, GCL+IPL, INL+OPL, ONL+IS, age, gender and axial length. Results The thickness of the OS+RPE layer was significantly decreased with the decrease of mTH (coefficient = 0.63 p

Purpose To create a method to identify the edge of visual field (VF) scotoma and to investigate the relationship between the steepness of the ‘border’ of scotoma and the prediction error of VF sensitivity. Methods Twenty-two glaucomatous patients with reliable VFs were recruited. Then, using the latest VFs (Humphrey 24-2) of these patients, the gradient of the plane on the hill of vision is calculated from the sensitivity of adjacent four/three points, so that the ‘border’ of VF damage is identified. Next, VF measurement was carried out, adding ten test points at the centre of adjacent four/three points where the gradient of the plane is largest (Full threshold, Custom mode). The VF measurements were performed using two approaches; random target presentation of 62 test points together and showing ten added points following or prior (randomly selected) to 52 (24-2) points. Each measurement with each approach was repeated twice in a same visit. Then, the absolute value of the difference between the measured sensitivity of the added ten test points and the average of the sensitivities of surrounding three/four test points were calculated. Finally, the relationship between the gradient of sensitivity plane and the absolute difference was investigated using the multilevel modeling (MLM). Results MLM revealed significant positive relationship between the absolute difference and the gradient of plane with all of the four measurements (p

A new isolation method of fungi was established by using the Percoll density gradient centrifugation. We found that the difference in the specific gravity allowed us to separate mixtures of fungal spores of Aspergillus, Fusarium, Listeromyces, Penicillium, and Trichoderma. When this technique was applied to soil samples, a wider variety of fungi were isolated compared with those obtained from an ordinary serial dilution method. For screening purposes, it is necessary to eliminate redundant strains from the strains initially isolated. For the efficient discrimination of strains, the random amplified polymorphic DNA (RAPD) method was applied. The establishment of a standard method for RAPD enabled us to minimize the deviation of Rf values between gels to less than 0.03. Conversion of the Rf values to binary matrices provided a matrix data that could be analyzed statistically. Based on the RAPD, it was possible to select genetically distinct fungal strains within a species. Key words: fingerprinting, isolation method, microbial screening, PCR, Percoll density gradient, RAPD.

Fungal strain NR 6356, Fusarium merismoides Corda, was discovered as the source of the protein kinase C (PKC) inhibitor, azepinostatin. The strain was identified based on its growth on potato sucrose agar, slender conidial shape, characteristic polyphialide and production of abundant chlamydospores. Fusarium aquaeductuum Lagh. IMI 103658 and Fusarium sp. NR 7222 were also found to produce the same inhibitor. After single colony isolation and medium optimization trials, a more than 30-fold increase in the production of azepinostatin over the original culture was achieved. Azepinostatin selectively and potently inhibited rat brain PKC with an IC50 value of 70 nM. Other enzymes utilizing ATP, including hexokinase, were not affected. The Ki of azepinostatin for PKC was 0.5 nM. The inhibition of PKC was competitive with ATP and uncompetitive with histone.

A novel phospholipase C inhibitor, hispidospermidin, was discovered from a fungal culture broth. The producing fungus, NR 7127, formed abundant pycnidia on banana leaf agar under near UV light. The ostiolate pycnidia were dark colored with a short beak possessing numerous protruding setae. The conidiogeneous cells were phialidic. The conidia were hyaline, 1 septate, smooth and spindle-shaped. From these distinctive characteristics, this strain was identified as Chaetosphaeronema hispidulum (Cda) Moesz of the Coelomycetes. Hispidospermidin was produced in a 50-liter jar fermentor containing 2% glucose, 2% potato starch, 2% Toast soya, 0.5% yeast extract, 0.25% NaCl, 0.0005% ZnSO4.7H2O, 0.0005% CuSO4.5H2O, 0.0005% MnSO4.4H2O, 0.32% CaCO3, and 0.3% Nissan disfoam CA-115. Fermentation was conducted at 27 degrees C at an aeration rate of 30 liters/minute and agitated at 500 rpm for 95 hours. Maximum production yield of hispidospermidin was observed after 72 hours. Hispidospermidin inhibited rat brain phospholipase C at 16 microM of IC50. This is the first recorded discovery of a secondary metabolite from the genus Chaetosphaeronema.

To investigate the relationship between methods used to evaluate the visual field (VF) and vision-related quality of life (VRQOL) in Japanese patients. We evaluated the VRQOL of 50 patients using the National Eye Institute Visual Function Questionnaire 25 (VFQ-25). Patients meeting the following criteria were assessed: (1) a Goldmann perimetry (GP) test conducted within 3 months of the initial visit; (2) best-corrected decimal visual acuity (VA) of the better eye when VRQOL was measured was less than 0.3 but more than 0.05. The VF was estimated based on a composite measure of the V-4e isopters of both eyes, as measured by GP. We calculated the VF using the American Medical Association (AMA) score, the Esterman disability score (EDS), the functional field score (FFS), the composite VF area calculated with area measurement software, and SR, an evaluation method based on solid angles. We then studied the relationships between the VFQ-25 score and the VF scores. Only EDS correlated significantly with the VFQ-25 score (r = 0.34, P = 0.02); AMA scoring did not correlate with the VFQ-25 score. It is necessary to reexamine standard VF evaluation methods.

To assess the prevalence of atypical retardation pattern (ARP) in scanning laser polarimetry (SLP) with variable corneal compensation (VCC) and enhanced corneal compensation (ECC), and to evaluate the factors relating to typical scan score (TSS). Measurements of SLP–VCC, SLP–ECC, and the Humphrey field analyzer (HFA) were performed in 105 normal subjects and 82 open-angle glaucoma (OAG) patients. ARP was defined as an SLP image with TSS

To compare the retinal nerve fiber layer thickness (RNFLT) parameters obtained with newly introduced GDx with enhanced corneal compensation (ECC) and those with GDx with variable corneal compensation (VCC) and to evaluate their reproducibility and the correspondence with visual field damage (VFD) in total or sectorized fields in association with refractive errors in open-angle glaucoma (OAG) patients.Measurement reproducibility was assessed in 30 normal and 30 OAG eyes. Correlation between the RNFLT parameters and the corresponding VFD was evaluated in 58 OAG eyes.All parameters of both GDx VCC and ECC showed high intraclass correlation coefficients among the repeated measurements (0.89 to 0.99), suggesting good reproducibility. All RNFLT parameters were significantly correlated between VCC and ECC (intraclass correlation coefficient=0.58-0.92, P0.001) though they were significantly different. In OAG eyes, correlation between temporal, superior, nasal, inferior, temporal average and mean deviation of VFD was similar in both algorithms (Rs=0.58 and 0.53, P0.001). When the OAG eyes were subgrouped by refractive error at -5 D, the correlation was significant for both ECC and VCC in the lower myopic group (-5 D) (Rs=0.71 and 0.74, P0.0001) but was significant only for ECC in the higher myopic group (or=-5 D) (ECC: Rs=0.44, P=0.016, VCC: Rs=0.19, P=0.309). In all 58 OAG eyes, significant correlation between the sectorized VFD and the corresponding RNFLT was found in the superior arcuate, inferior arcuate, and inferior para-arcuate areas for both VCC and ECC in the sector analysis (P0.01), whereas the correlation was significant only for ECC (Rs=0.46, P0.001) but not for VCC (Rs=0.30, P=0.024) in the superior para-arcuate.The results of GDx ECC were well correlated with those of VCC and had good reproducibility equivalent to that of VCC. Introduction of the ECC algorithm should improve the correspondence with VFD at least in moderate to high myopic OAG patients.

To evaluate the possible correlation between quality of life (QOL) characteristics and grades of legal visual impairment.Quality of life characteristics associated with various causes of visual loss were investigated in 237 Japanese patients in the low-vision care clinic at Tokyo University Hospital, who had been issued the official ID and handbook for the physically handicapped. The main causes of vision loss were glaucoma (87 patients), macular degeneration (35) and diabetic retinopathy (34). Using a previously developed questionnaire, we assessed the correlation between the disability index (DI) as a quality of life characteristic, and the grade of legal visual impairment.Total DI was 1.24 +/- 0.43, showing a very significant correlation with Handicapped Handbook grades (r = -0.44, p.0001). The correlation coefficients were -0.63 (p0.0001) for diabetic retinopathy, -0.50 (p = 0.002) for macular degeneration, and -0.16 (p = 0.14) for glaucoma patients.The needs of low-vision patients correlate with the grade of legal visual impairment and it is important to understand that the needs of low vision patients differ for each disease.

Objective: Trastuzumab (Tras) resistance is an important clinical problem to be resolved for HER2-positive gastric cancers (GC). Even after the tumor develops resistance to Tras, continuous anti-HER2 therapies, including Tras with chemotherapy (Tras beyond PD) or trastuzumab emtansine (T-DM1), could be effective treatment options. The Tras-sensitive cell line GLM-1 was established from a liver metastasis of a GC patient (Nakanishi et al. 2005), and the Tras-resistant cell line GLM1-HerR2 (R2) was established from GLM-1 by in vivo selection (Oshima et al. 2014). In this study, we used GLM1 and R2 xenograft models to examine the Tras-resistant mechanism of R2 and the effects of Tras plus paclitaxel (PTX) or T-DM1. Methods: BALB/c-nu/nu mice subcutaneously inoculated with GLM1 or R2 received Tras (40 mg/kg, qw) intraperitoneally, PTX (15 mg/kg, qw) or T-DM1 (10-40 mg/kg, q3w) intravenously (Day 1). Tumor volume (TV) was measured to evaluate antitumor effects, HercepTest® was used for HER2 status, and flow cytometry to analyze Tras-binding ability. Cell proliferation after knockdown of HER family receptors was measured by BrdU incorporation. Signal transduction was examined by Western blot for tumors excised on Days 2 and 5. Results: In contrast to GLM-1, R2 showed resistance to Tras treatment in xenograft experiments. There were no clear differences in HER2 score (3+), Tras-binding ability, p-HER2, p-HER3, p-AKT levels, or PIK3CA status (no hot spot mutation). Effects of gene knockdown of EGFR/HER2/HER3 in GLM-1 and R2 on cell proliferation were similar (ca. 10%/50%-60%/10% reduction, respectively). In GLM-1 tumors, Tras treatment decreased p-AKT, increased p27 protein, and decreased p-Rb, resulting in G1 cell cycle arrest, whereas in the R2 tumors changes in those molecules were few. These results suggest that escape from Tras-induced G1 arrest could be a resistant mechanism of R2. Based on the above results, and because in R2 cells HER2 remained a driver of proliferation and mitotic arrest was maintained, we hypothesized that Tras + PTX would show a higher antitumor effect than PTX alone. In the R2 xenograft model on Day 22, the Tras + PTX group showed a significantly higher antitumor activity (TV: 2 ± 3 mm3) than the PTX alone group (TV: 208 ± 209 mm3). In tumor specimens from the PTX and Tras + PTX groups, p-histon H3 (an M phase marker) was increased on Day 2. However, the Tras + PTX group, unlike the PTX group, showed markedly decreased p-histon H3 on Day 5, which suggests that Tras in combination with PTX enhances apoptosis in R2 tumors. Interestingly, T-DM1 showed dose-dependent antitumor activity in both GLM-1 and R2 xenografts. Conclusion: It was indicated that continuous anti-HER2 treatment, including Tras with chemotherapy or T-DM1 monotherapy, could be efficacious treatments for HER2-positive Tras-resistant GC tumors that emerge during previous Tras treatment. Citation Format: Sei Shu, Yoriko Yamashita-Kashima, Mieko Yanagisawa, Yoichiro Moriya, Naoki Harada. Role of trastuzumab in the combination treatment for a HER2-positive trastuzumab-resistant gastric cancer xenograft model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2691. doi:10.1158/1538-7445.AM2015-2691

Background: Irinotecan (CPT-11) and doxifluridine (5′-DFUR) are active agents against colorectal cancer. Each drug, however, has the possibility of causing diarrhea. Methods and Results: First, we determined the optimal dosing regimen in murine models. CPT-11 (i.v., q2d ×3) and 5′-DFUR (p.o., qd ×14) were administered to mice bearing a human colorectal cancer xenograft model. Diarrhea was stronger in the simultaneously administered schedule but not much stronger in the sequentially administered schedule compared with monotherapies. Both schedules yielded similar antitumor efficacies. Next, we conducted a phase I study combining CPT-11 on days 1 and 15, and 5′-DFUR on days 3–14 and 17–28 every 5 weeks in 19 patients with metastatic colorectal cancer. The doses of CPT-11 ranged from 80 to 150 mg/m2 and those of 5′-DFUR from 800 to 1,200 mg. Diarrhea of grade 3/4 developed in only 1 patient at 100 mg/m2/800-mg doses. Dose-limiting toxicities were hyperbilirubinemia and skipping doses due to fatigue at 150 mg/m2/1,200-mg doses. Conclusion: For the phase II study, the recommended dose was set at CPT-11 150 mg/m2 and 5′-DFUR 800 mg.

A combination therapy with CPT-11 and 5-FU/LV has been recently established as a first-line therapy for metastatic colorectal cancer. However, severe adverse effects have also been reported from this combination therapy, and a modality to reduce the adverse effects is desired. 5'-DFUR, a pro-drug of 5-FU, shows less myelotoxicity than 5-FU, and thus it may be a better partner to combine with CPT-11. However, since each drug has the possibility of inducing diarrhea, there is concern about their use in combination therapy. Therefore, in the present study, our aim was to establish an optimal schedule in murine models, which shows no increase in diarrhea but maintains potent antitumor activity. In non-tumor bearing mice, CPT-11 was given i.v. at 100 mg/kg/day q2d x 3, and 5'-DFUR was given p.o. at 172 mg/kg/day daily for 14 days. Each of these doses caused diarrhea in the single treatment. CPT-11 was administered simultaneously or sequentially with 5'-DFUR. With the simultaneously administered schedule, the diarrhea appeared stronger than that found in the CPT-11 single or in the 5'-DFUR single treatment groups. On the other hand, with the sequentially administered schedule the diarrhea was not much stronger than that found in the single agent treatment groups. When CPT-11 and 5'-DFUR administrations were separated by three-day intervals, the diarrhea was not augmented at all. In mice bearing human colorectal cancer COLO 205, the antitumor activity of CPT-11 in the combination with 5'-DFUR was additive in all of the examined schedules. The efficacy in the sequential schedule was the same as in the simultaneous schedule. These results suggest that a sequential administration schedule of CPT-11 and 5'-DFUR would be more tolerable than and equally efficacious to the simultaneous administration schedule. Clinical study of this sequential administration in combination therapy is warranted.

Purpose To develop and evaluate a method to map glaucomatous visual field (VF) damage by allocating additional test points to the standard 6° interval VF test pattern, considering the distribution of VF scotomata. Methods Subjects comprised 22 glaucomatous patients. Gradients of sensitivity were calculated for "squares" of test points in a patient's 24-2/30-2 VF results, so that the edges of scotomata could be identified where gradients were steep. Next, 10 new VF points were placed in these locations for each patient. Each patient's VF was then measured using this novel test grid (52 standard 24-2 test points and 10 additional points examined concurrently) on two separate occasions. The absolute difference between the measured sensitivity at each new additional test point and the average of the sensitivities of its surrounding four test points was calculated (Δave). The intra- and intervisit reproducibility of the additional test points' thresholds was calculated. Finally, fluctuation of overall VF damage was estimated using the intraclass correlation coefficient (ICC) and the coefficient of variation (CV). Results The average of the sensitivities (Δave) increased as the gradient of the plane steepened, whereas the reproducibility of the additional test points' thresholds remained stable. ICC was significantly higher and CV was significantly lower for the novel test grid compared with the standard 24-2 test pattern. Conclusions It may be advantageous to increase the density of VF test points where there are large local differences in VF sensitivity. These additional measurements may result in more reproducible and well-defined estimates of scotomata.

Background Bevacizumab is a humanized monoclonal antibody to human vascular endothelial cell growth factor (VEGF) and has been used for many types of cancers such as colorectal cancer, non-small cell lung cancer, breast cancer, and glioblastoma. Bevacizumab might be effective against gastric cancer, because VEGF has been reported to be involved in the development of gastric cancer as well as other cancers. On the other hand, there are no established biomarkers to predict the bevacizumab efficacy in spite of clinical needs. Therefore, we tried to identify the predictive markers for efficacy of bevacizumab in gastric cancer patients by using bevacizumab-sensitive and insensitive tumor models. Methods Nine human gastric and two colorectal cancer mouse xenografts were examined for their sensitivity to bevacizumab. We examined expression levels of angiogenic factors by ELISA, bioactivity of VEGF by phosphorylation of VEGFR2 in HUVEC after addition of tumor homogenate, tumor microvessel density by CD31-immunostaining, and polymorphisms of the VEGF gene by HybriProbe™ assay. Results Of the 9 human gastric cancer xenograft models used, GXF97, MKN-45, MKN-28, 4-1ST, SC-08-JCK, and SC-09-JCK were bevacizumab-sensitive, whereas SCH, SC-10-JCK, and NCI-N87 were insensitive. The sensitivity of the gastric cancer model to bevacizumab was not related to histological type or HER2 status. All tumors with high levels of VEGF were bevacizumab-sensitive except for one, SC-10-JCK, which had high levels of VEGF. The reason for the refractoriness was non-bioactivity on the phosphorylation of VEGFR2 and micro-vessel formation of VEGF, but was not explained by the VEGF allele or VEGF165b. We also examined the expression levels of other angiogenic factors in the 11 gastrointestinal tumor tissues. In the refractory models including SC-10-JCK, tumor levels of another angiogenic factor, bFGF, were relatively high. The VEGF/bFGF ratio correlated more closely with sensitivity to bevacizumab than with the VEGF level. Conclusions VEGF levels and VEGF/bFGF ratios in tumors were related to bevacizumab sensitivity of the xenografts tested. Further clinical investigation into useful predictive markers for bevacizumab sensitivity is warranted.

Objective: Triple-negative (ER-/PR-/HER2-) breast cancer (TNBC) is associated with a poor prognosis. Combination therapy with capecitabine (Cape) and bevacizumab (BV) has been reported to improve the progression-free survival of breast cancer patients (Robert NJ et al. ASCO 2009; RIBBON-1). In addition, Cape plus cyclophosphamide (CPA) combination therapy is considered to be effective for metastatic breast cancer patients (Tanaka M et al. Anticancer Drugs 2010). These findings prompted us to examine the antitumor efficacy of Cape+BV+CPA combination therapy in a TNBC xenograft model. Materials and Methods: MX-1 human TNBC cells were subcutaneously inoculated into the right flank of BALB-nu/nu mice. Treatment began (day 1) when tumor volume reached approximately 200 mm3. The mice were given Cape (359 mg/kg) and CPA (10 mg/kg) orally, 14 days on and 7 days off. BV (5 mg/kg) was administered intraperitoneally every 7 days for 3 weeks. Tumor volumes (mm3) were monitored twice a week and antitumor activity was evaluated on day 22. Six mice were allotted to each group (Cape+BV+CPA, Cape+BV, Cape+CPA, CPA, BV, control). Thymidine phosphorylase (TP: U/mg protein) levels in the tumor were measured by ELISA methods on day 15. Microvessel density (MVD, %) in tumor tissue was determined using CD31 immunohistochemical staining method on day 15. MVD% was calculated as the ratio of the CD31-positive staining area to the total observation area in the viable region. Results and Discussion: The antitumor activity of Cape+BV+CPA was significantly higher than that of Cape+BV or CPA (p≤0.05) and was also significantly stronger than that of Cape+CPA or BV (p≤0.05). The tumor volume on day 22 was as follows: control, 3969±1026; BV group, 2219±410; CPA group, 2149±962; Cape+BV group, 1270±458; Cape+CPA group, 1534±133; Cape+BV+CPA group, 536±261. The TP levels of the CPA (11.6±2.62), BV (6.21±2.59) and BV+CPA (13.1±2.70) groups were significantly increased compared to the control group (3.77±0.883, p≤0.05). The TP levels of the BV+CPA group were significantly higher than those of the BV group. (p≤0.05). The MVD% of BV (1.92±0.385), Cape+BV (1.26±0.347) and Cape+BV+CPA (1.00±0.160) was significantly decreased compared to the control group (3.57±0.343, p≤0.05). The MVD of the Cape+BV+CPA group was significantly depressed compared to that of the Cape+CPA (3.27±0.578), p≤0.05) group. These results indicate that 5-FU converted from Cape was increased by up-regulated TP with CPA, that it caused antitumor activity of Cape+BV+CPA superior to Cape+BV. BV significantly reduced MVD either alone or in the triplet combination (Cape+BV+CPA) and it is considered to result in the greatest effect in the triplet combination. Conclusion: Our preclinical results suggest that the Cape+BV+CPA (XAC) combination may be a rational therapy to be explored in TNBC patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4256. doi:10.1158/1538-7445.AM2011-4256

Our vision system receives two different images from our two eyes. Since these two images usually have high correlation they are fused into a single stable image. In binocular rivalry suppression a stimulus presented in one eye is suppressed and a different stimulus in the other eye is perceived for a certain time interval. For strabismic observers the suppression is continuous between different retinal images caused by misalignment of eye positions. Previous research investigated the effects of this binocular rivalry suppression on a number of visual processes. The present study looked at the effects of contrast adaptation for the adaptation stimulus presented to eyes under interocular suppression. We compared normal and strabismic observers and found that in both types, regardless of the existence of suppression, there was the effect of contrast adaptation. These findings provide evidence that the suppression occurs at a site in the visual system after the locus of contrast adaptation. The contrast adaptation strength differed in the two types of observers.