Exploratory open-label clinical study to determine the S-588410 cancer peptide vaccine-induced tumor-infiltrating lymphocytes and changes in the tumor microenvironment in esophageal cancer patients.

Cancer vaccines induce cancer-specific T-cells capable of eradicating cancer cells. The impact of cancer peptide vaccines (CPV) on the tumor microenvironment (TME) remains unclear. S-588410 is a CPV comprising five human leukocyte antigen (HLA)-A*24:02-restricted peptides derived from five cancer testis antigens, DEPDC1, MPHOSPH1, URLC10, CDCA1 and KOC1, which are overexpressed in esophageal cancer. This exploratory study investigated the immunologic mechanism of action of subcutaneous S-588410 emulsified with MONTANIDE ISA51VG adjuvant (median: 5 doses) by analyzing the expression of immune-related molecules, cytotoxic T-lymphocyte (CTL) response and T-lymphocytes bearing peptide-specific T-cell receptor (TCR) sequencing in tumor tissue or blood samples from 15 participants with HLA-A*24:02-positive esophageal cancer. Densities of CD8+, CD8+ Granzyme B+, CD8+ programmed death-1-positive (PD-1+) and programmed death-ligand 1-positive (PD-L1+) cells were higher in post- versus pre-vaccination tumor tissue. CTL response was induced in all patients for at least one of five peptides. The same sequences of peptide-specific TCRs were identified in post-vaccination T-lymphocytes derived from both tumor tissue and blood, suggesting that functional peptide-specific CTLs infiltrate tumor tissue after vaccination. Twelve (80%) participants had treatment-related adverse events (AEs). Injection site reaction was the most frequently reported AE (grade 1, n = 1; grade 2, n = 11). In conclusion, S-588410 induces a tumor immune response in esophageal cancer. Induction of CD8+ PD-1+ tumor-infiltrating lymphocytes and PD-L1 expression in the TME by vaccination suggests S-588410 in combination with anti-PD-(L)1 antibodies may offer a clinically useful therapy.Trial registration UMIN-CTR registration identifier: UMIN000023324.

Proton pump inhibitor after endoscopic resection for esophageal squamous cell cancer: multicenter prospective randomized controlled trial.

BACKGROUND: Whether proton pump inhibitors (PPIs) relieve heartburn or precordial pain after endoscopic resection (ER) for esophageal squamous cell carcinoma (ESCC) remains unclear. The aim of this study was to investigate the efficacy of PPI therapy for these symptoms after ER for ESCC. METHODS: We conducted a multicenter prospective randomized controlled trial among 15 hospitals in Japan. In total, 229 patients with cT1a ESCC were randomly assigned to receive PPI therapy for 5 weeks after ER (the PPI group, n = 115) or follow-up without PPI therapy (the non-PPI group, n = 114). The primary end point was the incidence of gastroesophageal reflux disease (GERD)-like symptoms after ER from a self-reported questionnaire (Frequency Scale for Symptoms of GERD). Secondary end points were ulcer healing rate at 5 weeks, incidence of pain, improvement rate of symptoms in those who started PPI therapy because of GERD-like symptoms in the non-PPI group, and adverse events. RESULTS: No significant difference was observed in the incidence of GERD-like symptoms after ER between the non-PPI and PPI groups (30 % vs 34 %, respectively). No significant differences were observed in the ulcer healing rate at 5 weeks (84 % vs 85 %) and incidence of pain within 1 week (36 % vs 45 %). In nine of ten patients (90 %) who started PPI therapy because of GERD-like symptoms in the non-PPI group, PPI administration relieved GERD-like symptoms. No adverse events related to PPI administration were observed. CONCLUSION: PPI therapy is not efficacious in reducing symptoms and did not promote healing of ulcers in patients undergoing ER for ESCC.

Serum prorenin levels and diabetic retinopathy in type 2 diabetes: new method to measure serum level of prorenin using antibody activating direct kinetic assay.

AIM: To investigate the serum levels of prorenin and its correlation with the severity of diabetic retinopathy (DR). METHODS: 248 patients with diabetes and 108 control subjects were divided into four groups: no-DR (n = 146), no proliferative diabetic retinopathy (no-PDR) (n = 78), PDR (n = 24), and controls (n = 108). Serum levels of prorenin from all subjects were measured using the new antibody activating direct kinetic (AAD-PR) assay. The serum prorenin levels were compared among the groups. RESULTS: The serum levels of prorenin in the control, no-DR, no-PDR, and PDR groups, respectively, were 109.1 (66.1), 194.6 (160.4), 271.5 (220.3), and 428.4 (358.4) pg/ml (mean (SD)). Prorenin in the PDR group was remarkably high compared with the control and no-DR groups (p<0.0001) and with the no-PDR group (p = 0.002). Serum levels of prorenin increased with increasingly severe retinopathy. No correlation was found between the prorenin level and the duration of disease or HbA(1c). CONCLUSIONS: The serum levels of prorenin in patients with PDR were found to be markedly high using the AAD-PR assay. Increased levels of prorenin in diabetes may have an important role in the pathogenesis of DR.

Peroxynitrite decomposition catalyst, FP15, and poly(ADP-ribose) polymerase inhibitor, PJ34, inhibit leukocyte entrapment in the retinal microcirculation of diabetic rats.

PURPOSE: Oxidative and nitrosative stress and activation of poly(ADP ribose) polymerase (PARP) play a role in the pathogenesis of diabetic complications. We evaluated the effectiveness of the peroxynitrite decomposition catalyst, FP15, and the PARP inhibitor, PJ34, in the treatment of leukocyte entrapment in the retinal microcirculation of diabetic rats. METHODS: Diabetes was induced in rats by intraperitoneal injection of 60 mg/kg of streptozotocin. Rats were divided into four groups: controls; untreated diabetes; diabetes treated with FP15 (10 mg/kg oral gavage twice daily) and diabetes treated with PJ34 (10 mg/kg oral gavage twice daily). All experiments were performed 4 weeks after initiation of treatment. Leukocyte entrapment in the retinal microcirculation was quantitatively evaluated in vivo with acridine orange digital fluorography. RESULTS: The density of leukocytes trapped in the retinal microcirculation 30 minutes after dye injection was significantly greater in untreated diabetes (32.1 +/- 4.7 cells/mm2) than in controls (11.3 +/- 4.5 cells/mm2) (p < 0.05). Compared with untreated diabetes, the density of trapped leukocytes significantly decreased in diabetes treated with FP15 (14.5 +/- 5.1 cells/mm2) (p < 0.0001) and diabetes treated with PJ34 (24.1 +/- 4.2 cells/mm2) (p < 0.05). CONCLUSIONS: Treatment with FP15 and PJ34 decreased enhanced leukocyte entrapment in the retinal microcirculation during the early diabetic period. The current study suggests a role for peroxynitrite production and for PARP activation in the pathogenesis of retinal microvascular leukostasis in early diabetes.

Alteration of choroidal circulation in the foveal region in patients with type 2 diabetes.

AIM: To investigate changes in choroidal blood flow (CBF) in the foveal region in patients with type 2 diabetes. METHODS: Laser Doppler flowmetry was used to determine the CBF in the foveal region in 70 patients with type 2 diabetes and 36 age and sex matched healthy subjects (control group). The patients were classified into three groups: 33 patients (33 eyes) with no diabetic retinopathy (NDR), 20 patients (20 eyes) with non-proliferative diabetic retinopathy and no macular oedema (NPDR/MO-), and 17 patients (17 eyes) with NPDR and MO (NPDR/MO+). Optical coherence tomography was also used to measure the foveal thickness. RESULTS: The group averaged CBF values were 13.5 (4.9), 9.4 (2.5), 10.8 (4.8), and 5.6 (2.0) (arbitrary units) in the control, NDR, NPDR/MO-, and NPDR/MO+ groups, respectively. The group averaged CBF values in the NDR group decreased (30.2%; p<0.01) compared with the control group. The average CBF value in the NPDR/MO+ group was also significantly lower (48.2%; p<0.01) compared with that in the NPDR/MO- group. CONCLUSION: The CBF in the foveal region significantly decreases in patients with diabetes, especially those with macular oedema.

Features of abnormal choroidal circulation in central serous chorioretinopathy.

AIMS: To evaluate abnormalities in the choroidal circulation in cases of central serous chorioretinopathy (CSC). METHODS: A complete clinical ophthalmological examination was performed using simultaneous fluorescein and indocyanine green (ICG) angiography with a confocal scanning laser ophthalmoscopy and the digital images analysed in 36 consecutive patients with acute CSC. To quantify the choroidal circulation, the foveal choroidal blood flow was measured in 11 patients using laser Doppler flowmetry. RESULTS: Fluorescein angiography showed focal leakage from the retinal pigment epithelium in all patients. ICG angiography revealed delays in arterial filling in 27 eyes (75%), and fluorescein angiography showed small hypofluorescent points around the leakage in 27 eyes (75%). Abnormal choroidal hyperfluorescence was observed in 30 eyes (83%). The choroidal blood flow in eyes with CSC was 45% lower than in fellow eyes (p<0.01). CONCLUSION: Decreased choroidal blood flow in CSC was demonstrated for the first time. The decreased choroidal blood flow might be correlated with the small, localised hypofluorescent areas, which may indicate non-perfused areas of the choriocapillaris that are frequently seen during ICG angiography.

Inhibitory effect of losartan, an AT1 angiotensin II receptor antagonist, on increased leucocyte entrapment in retinal microcirculation of diabetic rats.

BACKGROUND: The effectiveness of losartan for the treatment of leucocyte entrapment in the retinal microcirculation of diabetic rats was evaluated quantitatively. METHODS: After diabetes was induced by injection of streptozotocin (STZ), the rats were divided into two subgroups. The first subgroup (n = 6), received no medications; the second subgroup (n = 6) was given fresh drinking water supplemented with losartan (5 mg/kg/day) for 4 weeks. Six rats that were not injected with STZ or given medications served as controls. 4 weeks after intervention, leucocyte dynamics in the retina were observed using acridine orange digital fluorography. Leucocyte entrapment in the retina was compared among the three groups. RESULTS: In the untreated diabetic rats, the number of trapped leucocytes (6.1 (SD 1.4) cells/mm(2)) increased significantly compared with control rats (2.8 (1.2) cells/mm(2); p = 0.005) and diabetic rats treated with losartan (3.1 (0.9) cells/mm(2); p = 0.0002). CONCLUSIONS: Losartan, an AT1 angiotensin II receptor antagonist, inhibited increased leucocyte entrapment in the diabetic retina. The authors demonstrated that losartan may have therapeutic efficacy in preventing development of diabetic retinopathy. Further clinical studies of the effect of the angiotensin receptor antagonist on preventing development of diabetic retinopathy are needed.

Scotoma and fixation patterns using scanning laser ophthalmoscope microperimetry in patients with macular dystrophy.

PURPOSE: We used scanning laser ophthalmoscope microperimetry to evaluate the retinal scotoma and the fixation points in the patients with macular dystrophy. METHODS: We studied 10 eyes of five patients with macular dystrophy (three patients with cone dystrophy and two patients with Stargardt disease). The mean patient age was 37 years (range, 13 to 64 years). An estimation of scotoma and fixation points on the retina was performed using scanning laser ophthalmoscope microperimetry. RESULTS: All 10 eyes (100%) had one of two types of dense scotoma: type one was a dense ring scotoma (five eyes, 50%), and type two was a dense central scotoma (five eyes, 50%) that included the center of the fovea. In all eyes with a dense ring scotoma, the fixation points were stable and did not shift. In all eyes with a dense central scotoma, the fixation shifted. The logarithm of minimal angle of resolution of the visual acuity in the eyes with the dense central scotoma was significantly worse than that of eyes with the dense ring scotoma type (P =.005). CONCLUSIONS: Scanning laser ophthalmoscope microperimetry findings demonstrate two types of dense scotoma (dense ring scotoma and dense central scotoma) in the patients with macular dystrophy. The two types of dense scotoma affect the shifting of the fixation points and the stability of fixation and may result in the difference in visual acuity in the patients with macular dystrophy.

[Immunohistochemical study of idiopathic and secondary epiretinal membrane].

PURPOSE: We investigated the immunohistochemical features of surgically resected idiopathic epiretinal membranes(ERMs) and secondary ERMs with regard to posterior vitreous detachment(PVD). METHODS: Six specimens of idiopathic epiretinal membranes(3 eyes with complete PVD, 2 eyes with partial PVD, and one eye with no PVD) and 3 specimens of secondary ERMs(all eyes with complete PVD) were immunohistochemically studied. We used type I, II, III, IV collagen and fibronectin to study extracellular components, and glial fibrillary acidic protein(GFAP), S 100 protein, vimentin, and so forth to study cellular components. RESULTS: All the specimens of idiopathic ERMs had the major components of the lamellar stained by type II collagen antibody, and one out of 3 specimens of secondary ERMs had a minor component stained by type II collagen antibody. Compared with idiopathic ERMs with complete PVD, 2 out of 3 specimens of idiopathic ERMs with partial PVD or no PVD contained rather thick collagen lamellar. CONCLUSION: There was difference between specimens of idiopathic ERMs and specimens of secondary ERMs in staining by type II collagen antibody, supposed by vitreous, in this study. Idiopathic ERM with attached posterior vitreous membrane may cause growth of collagen.

Inhibitory effect of losartan on laser-induced choroidal neovascularization in rats.

PURPOSE: To investigate the inhibitory effects of losartan, an angiotensin receptor antagonist, on angiogenesis in a rat model of laser-induced choroidal neovascularization. METHODS: Experimental study. Fifteen Brown-Norway male rats received losartan (approximately 5 mg/kg/d) in drinking water, and 15 Brown-Norway male rats received unsupplemented drinking water 1 week before photocoagulation, and it was continued to the end of the study. Two weeks after intense laser photocoagulation, choroidal neovascularization was evaluated by fluorescein angiography and histopathologic evaluation. RESULTS: The incidence of choroidal neovascularization formation was 99.5 +/-.2% (mean +/- standard deviation) in controls and 72.5 +/- 8.8% in losartan-treated rats (P <.01). Quantitative morphometric assessment revealed mean choroidal neovascularization lesion thickness of 54 and 44.8 microm, respectively, in controls and losartan-treated rats (P <.01). CONCLUSION: Losartan seems to inhibit development of laser-induced choroidal neovascularization. Angiotensin receptor antagonists may be useful as prophylaxis against choroidal neovascularization associated with age-related macular degeneration.

Involvement of RNase G in in vivo mRNA metabolism in Escherichia coli.

BACKGROUND: Escherichia coli rng gene (previously called cafA) encodes a novel RNase, named RNase G, which is involved in the 5' end-processing of 16S rRNA. In rng mutant cells, a precursor form of 16S rRNA, 16.3S rRNA, is accumulated. Here we report a role of RNase G in the in vivo mRNA metabolism. RESULTS: We found that rng:cat mutant strains overproduced a protein of about 100 kDa. N-terminal amino acid sequencing of this protein showed that it was identical to the fermentative alcohol dehydrogenase, the product of the adhE gene located at 28 min on the E. coli genetic map. The level of adhE mRNA was significantly higher in the rng:cat mutant strain than that in its parental strain, while such differences were not seen in other genes we examined. A rifampicin-chase experiment revealed that the half-life of adhE mRNA was 2.5-fold longer in the rng:cat disruptant than in the wild-type. CONCLUSION: These results indicate that, in addition to rRNA processing, RNase G is involved in in vivo mRNA degradation in E. coli.

Stereo acuity in patients with unilateral macular hole and after unilateral macular hole surgery.

PURPOSE: To investigate stereo acuity levels in patients with unilateral idiopathic macular hole and after surgical intervention. METHODS: In 31 consecutive patients with a unilateral macular hole and 46 consecutive patients who underwent successful unilateral macular hole surgery, complete ocular examinations, including orthoptic examinations and microperimetry using the scanning laser ophthalmoscope, were performed. RESULTS: A significantly positive correlation was found between VA and stereo acuity (r = 0.87, P < 0.01). After successful surgery, stereo acuity also correlated with the presence or absence of absolute and/or relative scotoma, and was best in eyes without scotomata. Patients with unilateral idiopathic macular hole, suppression, and symptom duration of 24 months or longer had no stereoscopic vision. CONCLUSIONS: The results indicated that in patients with unilateral idiopathic macular hole and after surgery, stereo acuity correlated with VA. Patients with unilateral macular hole should be operated upon as early as possible, resulting in better VA and better stereo acuity.

Dynamic observation of selective accumulation of a photosensitizer and its photodynamic effects in rat experimental choroidal neovascularization.

PURPOSE: The authors investigated the selective accumulation of a photosensitizer, ATX-S10(Na), in experimental choroidal neovascularization (CNV) in rats using a highly sensitive colorchromatic charge coupled device (CCD) camera. METHODS: To detect the development of experimental CNV in 30 rats, the animals were followed weekly with simultaneous fluorescein and indocyanine green angiography. After injecting ATX-S10(Na), the authors detected fluorescence from the photosensitizer using a highly sensitive color CCD camera. The camera was connected to a surgical microscope, under which rat fundi were observed through a coverglass in contact with the cornea. The retinas were excited with 405-435 nm light, and the light emitted from the photosensitizer passed through a 680-nm bandpass filter before being detected by the CCD camera. RESULTS: Immediately after injection, fluorescence appeared in the retinal vessels and then the entire retina. Thirty minutes postinjection, the intensity of the fluorescence was still strong from the whole retina, and the CNV was not detected. One hour after injection, retinal fluorescence was weak but still observable; 1.5 hours postinjection, retinal fluorescence was undetectable but fluorescence was strong from the CNV. Under the optimum therapeutic conditions, CNV was effectively occluded. CONCLUSION: ATX-S10(Na) selectively accumulates in the CNV in rats. The optimum therapeutic timing is approximately 1.5 hours postinjection of the dye in this CNV model.

New graphics models for PC based ocular surgery simulator.

High-end graphics workstations (GWS) have been used for surgical simulators utilizing Computer Graphics (CG) and Virtual Reality (VR) technologies. This is because the simulators need lots of computing power, mainly for collision detection among objects modeled as a set of polygons. In this paper, we propose to use mathematical functions to model objects for collision detection. However, for graphic display we continue to use polygonal representation. Using the new model, we have developed a PC based ocular surgery simulator, which creates realistic surgery image in real-time. The computation time was found to be much lower than that in the conventional method.

Pulsatile ocular blood flow study: decreases in exudative age related macular degeneration.

BACKGROUND: Pulsatile ocular blood flow (POBF) is a parameter for evaluating choroidal blood flow. POBF in the patients with non-exudative and exudative age related macular degeneration (AMD) was investigated. METHODS: POBF, pulse amplitude (PA), systolic and diastolic blood pressures, intraocular pressure (IOP), refractive error, and axial length were compared among 10 patients with non-exudative AMD, 11 patients with exudative AMD, and 69 age matched controls. A Langham OBF computerised tonometer was used with the participants in the sitting position to measure POBF and PA. RESULTS: No significant differences were found in age, systolic and diastolic blood pressures, IOP, or refractive error between patients with exudative and non-exudative AMD and the control subjects. In the patients with exudative AMD the POBF (median, 372.7 microl/min) and PA (median, 1.2 mm Hg) were significantly lower than in the patients with non-exudative AMD (median, 607.0 microl/min (p = 0.02) and 2.2 mm Hg (p = 0.04), respectively) and control subjects (median, 547.4 microl/min (p = 0.01) and 2.0 mm Hg (p = 0.01), respectively). CONCLUSIONS: These data show that the POBF and PA in the patients with exudative AMD are lower than in the patients with non-exudative AMD and normal subjects. Decreased choroidal blood flow may have a role in the development of choroidal neovascularisation in AMD.