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PURPOSE To compare the efficacy and safety of a combination therapy of prednisolone and cyclosporine and corticosteroid pulse therapy in Vogt-Koyanagi-Harada (VKH) disease. STUDY DESIGN A prospective, multicenter, randomized, non-inferiority trial. METHODS Patients of new-onset acute VKH disease at 11 centers in Japan between 2014 and 2018 were randomized to a combination (oral prednisolone 60 mg daily with gradual taper-off to 35 mg/day and cyclosporine 3 mg/kg/day) and corticosteroid (methylprednisolone 1000 mg for 3 days followed by oral prednisolone) groups, and were followed for 1 year. RESULTS Thirty-four were assigned to the combination and thirty-six patients to the corticosteroid group. Recurrence/worsening risk was 0.15 (95% confidence-interval [CI] 0.03-0.27) in the combination group and 0.25 (95% CI 0.11-0.39) in the corticosteroid group, with a risk difference of - 0.10 (90% CI - 0.27 to 0.06), demonstrating non-inferiority of the combination group with a non-inferiority margin of 0.20 (P = 0.0013). Serious adverse events occurred in three patients (two with hyponatremia and one with severe headaches) in the combination group and none in the corticosteroid group. Sunset glow fundus grades and cataract rates at 1 year were 0.57 (95% CI 0.42-71) and 4.3% in the combination group and 0.91 (95% CI 0.78-1.04) and 34.0% in the corticosteroid group, respectively. CONCLUSIONS Combination therapy was noninferior to corticosteroid therapy with respect to recurrence/worsening risk. Notably, the recurrence/worsening risk, sunset glow fundus grade, and cataract rate were lower in the combination group than in the corticosteroid group.

To report the clinicopathological findings of retinal vasoproliferative tumor/reactive retinal astrocytic tumor (VPT/RRAT) with retinal vasculitis, treated by tumor resection.A retrospective single case report.A 29-year-old Japanese woman was referred with cystoid macular edema and retinal vasculitis in the right eye. Best-corrected visual acuity (BCVA) was 0.9. Fundus examination, optical coherence tomography, and fluorescein angiography demonstrated VPT/RRATs in the temporal retina surrounded by a subretinal exudate, serous retinal detachment and macular edema, and retinal vasculitis. Despite 3 months of oral prednisolone treatment, a full-thickness macular hole developed. Pars plana vitrectomy and endoresection of the VPT/RRATs were performed. Pathological and immunohistochemical analyses with anti-CD34 antibody, anti-glial fibrillary acidic protein (GFAP) antibody, anti-Ki67 antibody, and anti-vascular endothelial growth factor (VEGF) antibody were conducted on the excised tissue. Inflammation was evaluated by immunohistological staining with leucocyte common antigen (LCA), anti-CD3 antibody, and anti-CD20 antibody.After surgery, the macular hole closed, BCVA improved to 1.2, retinal vasculitis was ameliorated, and retinal exudate disappeared. There was no recurrence of VPT/RRAT or retinal vasculitis. Pathological examination showed that GFAP and VEGF were widely expressed, irrespective of the distribution of blood vessels. Ki67-positive proliferating cells were detected in the perivascular area. LCA-positive leukocytes and CD3-positive T cells were detected throughout the samples, whereas CD20-positive B cells were rarely detected.Endoresection of VPT/RRAT could be a good treatment option for secondary VPT/RRAT accompanied by retinal vasculitis. Pathological findings revealed for the first time that inflammatory cells infiltrate the tissue in secondary VPT/RRAT.

Vitreoretinal lymphoma (VRL) is a subtype of diffuse large B-cell lymphoma and is sight- and life-threatening in the vast majority of patients. Lymphoma cells infiltrate the vitreous body and/or subretinal space and exhibit clinical signs of vitreous opacities and creamy white subretinal lesions. Although the intraocular signs can serve as clues to suspect VRL, they are nonspecific and may be misdiagnosed as uveitis. Histopathological evidence of malignant cells on vitreous biopsy, for instance, is the gold standard for diagnosis of VRL; however, cytological examination of the vitreous often results in a low success rate owing to the small quantity and poor quality of tissues and cells in the sample. Recent advancements in immunological, molecular, and gene analyses using intraocular samples have made it possible to accurately diagnose VRL. As for the management of VRL, local treatments with irradiation and/or intravitreal injections of anti-tumor agents (methotrexate or rituximab) are effective in suppressing intraocular VRL lesions. However, the effect of systemic chemotherapy, with or without brain irradiation, on preventing central nervous system involvements remains controversial. In this review article, we discuss the following concepts based on previous literature and our unpublished results: current ocular imaging examinations such as optical coherence tomography and fundus autofluorescence; immunological, molecular, and gene expression characterization of intraocular biopsies with special attention to flow cytometry; immunoglobulin gene rearrangement assays that use the polymerase chain reaction test; cytokine assays; gene mutations (MYD88, CD79B); and current local and systemic treatments of VRL.

Two patients who showed transient lesions in the splenium of the corpus callosum (SCC) secondary to acute ischemic stroke are reported. Both patients had embolic strokes and showed an isolated lesion in the SCC on magnetic resonance imaging (MRI) 1-2 weeks after the onset of stroke, with a hyperintense lesion on diffusion-weighted imaging and decreased apparent diffusion coefficient values, with no symptoms related to the lesion. In both cases, the lesion disappeared on MRI approximately 1 week later. Clinicians should note that transient SCC lesions can occur following acute ischemic stroke and avoid misdiagnosing them and performing unnecessary examinations or treatment.

Infliximab (IFX) was the first biologic introduced for refractory uveitis treatment in Behçet’s syndrome (BS). However, there have been few reports on the safety and efficacy of IFX monotherapy over follow-up periods of more than 10 years. This retrospective study evaluated the 10-year safety and efficacy of IFX monotherapy compared to IFX combination therapies with colchicine or corticosteroid for refractory uveitis in BS patients. Monotherapy was performed in 30 eyes of 16 patients while combination therapies were performed in 20 eyes of 11 patients. Continuation of IFX occurred in 70.3% of enrolled patients for 10 years without any significant difference noted in the retention rate between the monotherapy and combination therapies (p = 0.86). Reduction of ocular inflammatory attacks and improvement of best corrected visual acuity occurred in the monotherapy group after 10 years, which was equivalent to that for the combination therapies. Although adverse events (AEs) or therapy discontinuation occurred during the initial 5 years in both therapies, no AEs were observed for either therapy after 6 years. Our results suggested that IFX monotherapy proved to be effective and not inferior to combination therapies over a 10-year follow-up. Although loss of response and AEs may be noticed during the initial 5-year period, a safe and effective continuation can be expected thereafter.

Diagnosis of intraocular lymphoma is difficult. Among the hurdles in the diagnosis are the variety of reactive inflammatory and ischemic changes among intraocular lymphoma patients. Thus, a novel diagnostic method is desired such that lymphoma cells can be distinguished by the signals intrinsic to the cells, not by those from the surrounding tissues with reactive changes. Raman spectroscopy is a technique that can detect intrinsic signals from each cell. Therefore, Raman spectroscopy is a good candidate for an intraocular evaluation technology that could contribute to improve the diagnosis of intraocular lymphoma. In this study, we tested whether the intrinsic Raman signals from malignant lymphoma cells, in the absence of surrounding tissue, were sufficient for the discrimination of malignant lymphoma cells from leukocytes. We acquired spectra from dissociated lymphoma cells, along with spectra from normal B cells and other leukocytes involved in intraocular inflammatory diseases. We analysed the spectra using principal component analyses and quadratic discriminant analyses. We found that Raman spectra from dissociated cells without confounding tissues showed high discriminating ability, regardless of the variation due to day-to-day differences and donor differences. The present study demonstrates the possible effectiveness of Raman spectroscopy as a tool for intraocular evaluation.

Recently, we successfully transplanted an autograft, or major histocompatibility complex (MHC)-matched allografts, from induced-pluripotent-stem-cell-derived retinal pigment epithelial (iPSC-RPE) cells in patients with age-related macular degeneration. However, there was an issue regarding immune rejection after transplantation. In this study, we established a preoperational in vitro &ldquo, drug&ndash, lymphocytes&ndash, grafts immune reaction (Drug-LGIR)&rdquo, test to determine the medication for immune rejection using host immunocompetent cells (lymphocytes) and transplant cells (target iPSC-RPE cells) together with different medications. The adequacy of the test was assessed by in vivo transplantation in monkey models together with medication based on in vitro data. In the results of Drug-LGIR tests, some drugs exhibited significant suppression of RPE cell-related allogeneic reactions, while other drugs did not, and the efficacy of each drug differed among the recipient monkeys. Based on the results of Drug-LGIR, we applied cyclosporine A or local steroid (triamcinolone) therapy to two monkeys, and successfully suppressed RPE-related immune rejections with RPE grafts, which survived without any signs of rejection under drug administration. We propose that our new preoperational in vitro Drug-LGIR test, which specifies the most efficacious medication for each recipient, is useful for controlling immune attacks with personalized treatment for each patient after retinal transplantation.

Summary Antibody-mediated rejection is characterized by donor-specific antibody produced by B cells. However, to our knowledge, B cell invasion and antibody in the inflamed retina after transplantation of retinal pigment epithelial (RPE) cells has not been reported. To determine if RPE transplantation could be performed using allografts, we established in vivo immune rejection models with induced pluripotent stem cell (iPSC)-RPE allografts and determined whether RPE-specific antibody could be detected in these models. We detected alloantibodies in the serum from recipient monkeys that had immune attacks in the retina in an immunofluorescent assay using the transplanted iPSC-RPE cells as the antigen. In addition to T cell and antigen-presenting cell immunity, peripheral blood cells and lymph nodes in animal models with allogeneic iPSC-RPE cells also had activated B cells, which were probably secreting alloantibodies. Using serum and transplanted cells, alloreactive antibody can be detected for the diagnosis of immune rejection after transplantation., Graphical Abstract, Highlights • Alloantibodies secreted by B cells target iPSC-derived RPE cells • Alloantibodies were detected in serum from monkeys receiving RPE allografts • Blood and lymph nodes in monkeys receiving the grafts contained activated B cells • B cell-mediated immune attack was attenuated by administration of local steroid, In this article, Sugita and colleagues show that B cells, as well as T cells, are associated with immune attack on iPSC-derived retinal pigment epithelial cells (RPE) after allogeneic transplantation. In addition, it shows that B cells can produce alloantibodies against iPSC-RPE cells.

Objective To assess the correlation between the angiographic appearance of cerebral collateral pathways or the degree of internal carotid artery stenosis (ICAS) and reduced cerebrovascular reactivity (CVR) estimated by single-photon emission computed tomography (SPECT) image analysis in patients with unilateral ICAS. Methods A retrospective analysis was performed in 42 patients with unilateral ICAS who underwent cerebral angiography and acetazolamide-challenged SPECT of the brain. Cerebral blood flow quantitation was performed using the quantitative SPECT/dual-table autoradiography method. The CVR in the middle cerebral artery (MCA) territory was evaluated using the stereotactic extraction estimation based on the Japanese extracranial-intracranial bypass trial (SEE-JET) program and classified as reduced (

Recently, a new assay method that can quantify the singlet oxygen absorption capacity (SOAC) of natural antioxidants and food extracts was developed. The SOAC values were measured in ethanol–chloroform–D2O (50 + 50 + 1, v/v/v) solution at 35°C using a UV-Vis spectrophotometer equipped with a six-channel cell positioner and an electron-temperature control unit. In the present study, measurement of the SOAC values was performed for eight representative carotenoids and three vegetable extracts (tomato, carrot, and red paprika) using a versatile instrument, the microplate reader. A 24-well glass microplate was used for measurements because a plastic microplate, commonly used in the laboratory, dissolves in the ethanol–chloroform–D2O solution. The SOAC values of eight carotenoids and three vegetable extracts measured using a microplate reader were in good agreement with the corresponding values measured using a UV-Vis spectrophotometer, suggesting that the microplate reader is an applicable instrument for the measurement of reliable SOAC values for general antioxidants and food extracts in solution.

The retinal pigment epithelium (RPE) is essential for maintaining retinal homeostasis by removing and recycling photoreceptor outer segment (POS) in membranes. It also produces and secretes growth factors involved in retinal homeostasis. Arrestin 1 (ARR1) is specifically expressed in photoreceptors (PRs) and a vital molecule for keeping visual cycle between PRs and RPE. In the present study, we showed the expression of ARR1 was decreased by form-deprivation (FD) in retina of rat. The ARR1 was detected in the RPE of the controls but not in the RPE of FD, which indicates RPE phagocytes POS containing ARR1. Furthermore, we overexpressed ARR1 in cultured human RPE and revealed the ARR1 upregulates bFGF expression and downregulates TGF-β1, -β2 and bone morphogenetic protein-2 (BMP-2). The upregulation of bFGF by ARR1 directly works for PR survival and the downregulation of TGF-βs by ARR1 inhibits epithelial mesenchymal transition (EMT) of RPE, which is the underlying mechanism of keeping retinal homeostasis. Our results also indicate the regulation of ARR1 expression in RPE might become a novel therapeutic option for various ocular diseases., Highlights • Arr1 is predominantly expressed in photoreceptors. • The expression of ARR1 is decreased by form-deprivation in the RPE. • ARR1 altered bFgf and Tgf-β in the RPE, which have roles in retinal homeostasis.

Purpose To determine whether human induced pluripotent stem (iPS) cell-derived retinal pigment epithelial (RPE) cells (iPS-RPE) can suppress natural killer (NK) cell activation. Methods iPS-RPE cells were cocultured with peripheral blood mononuclear cells (PBMCs) or purified NK cells from healthy donors after stimulation with cytokines. To confirm expression of NK cell-specific markers, flow cytometry and quantitative RT-PCR (qRT-PCR) were performed. NK cells (or PBMCs) cocultured with iPS-RPE cells were assessed for proliferation by Ki-67 expression with flow cytometry, and NK suppression by RPE cells was assessed for granzyme B production with ELISA. Human leukocyte antigen (HLA) expression including HLA-E on iPS-RPE cells was evaluated with flow cytometry and qRT-PCR. The effect of HLA-E downregulation was also investigated using small interfering RNA (siRNA) systems. Following iPS-RPE cell transplantation in vivo, we evaluated NK cell invasion in the retina with immunohistochemistry. Results Activated NK cells expressed NK-related markers such as CD16, CD56, and CD11b, and NK cells produced cytotoxic agents such as granzyme B, perforin, and TNF-α. Human iPS-RPE cells inhibited cell proliferation and production of these cytotoxic agents by activated NK cells in vitro. iPS-RPE cells constitutively expressed HLA-E and suppressed NK cell activation through an interaction between HLA-E and CD94/NKG2A. Moreover, immunohistochemical evaluation of monkey RPE transplantation into in vivo immune rejection models showed no NK cell invasion in the retina in allografts or xenografts except for one xenografted eye. Conclusions Cultured iPS cell-derived RPE cells greatly suppress NK cell activation. Thus, NK cells might be inactivated when exposed to this type of retinal cell.

As for the risk management of the financial institution, a change is demanded to risk permission based on risk appetite from the risk aversion corresponding to the bad-loan problem. The financial institutions which performed risk management based on VaR (value at risk) fell into a situation of the management aggravation by the outbreak of the global financial crisis. They used an index to measure based on a past price change statistically. Many local financial institutions work on "business evaluation" included there since 2014. In 2016 Financial Services Agency demand "an evaluation (= business evaluation) depending on growth possibility of the business of the client appropriately without depending on a security, a guarantee excessively for a financial institution". In this way, policy demands switching the financing posture that came to the local financial institution depending on a security, a guarantee to the financing based on the business evaluation" Authors suggest a scheme to evaluate business value and risk and connect with the financing for the new risk that is hard to become an insufficient measurement with the conventional risk management model. In this study, authors analyzed the structure of the risk management when financial institution financed by business evaluation and decided the evaluation of the risk from the influence of the factor of the individual risk, and showed decision-making model

PURPOSE The purpose of this study was to determine whether human retinal pigment epithelial (RPE) cells from induced pluripotent stem (iPS) cells could inhibit T-cell activation in vitro. METHODS Cultured iPS-derived RPE (iPS-RPE) cells were established from fresh skin tissues or dental pulp cells obtained from healthy donors or a retinal patient after informed consent was obtained. To confirm expression of the specific markers on iPS and iPS-RPE cells, immunohistochemistry, quantitative RT-PCR (qRT-PCR), and flow cytometry were performed. Target T cells were obtained from peripheral blood mononuclear cells of healthy donors. Target T cells were assessed for proliferation by incorporation of bromodeoxyuridine or carboxyfluorescein succinimidyl ester for production of cytokines such as IFN-γ. Expression of TGFβ and other candidate molecules by iPS-RPE cells was evaluated with flow cytometry, ELISA, multiplex cytokine array, immunohistochemistry, and qRT-PCR. RESULTS The RPE cells we established from iPS cells had many characteristics of mature RPE cells but no characteristics of pluripotent stem cells. Cultured iPS-RPE cells inhibited cell proliferation and production of IFN-γ by activated CD4(+) T cells. In some bystander T cells, iPS-derived RPE cells induced CD25(+)Foxp3(+) regulatory T cells in vitro. Induced pluripotent stem-RPE cells constitutively expressed TGFβ and suppressed activation of T cells via soluble TGFβ, because TGFβ-downregulated iPS-RPE cells did not inhibit this T-cell activation. CONCLUSIONS Cultured iPS-derived retinal cells fully suppress T-cell activation. Transplantation of iPS-RPE cells into the eye might be a therapy for retinal disorders.

Background Myelin oligodendrocyte glycoprotein (MOG) antibody-positive optic neuritis (ON) and myelitis are recognized as important differential diagnosis of aquaporin-4 (AQP4) antibody-positive neuromyelitis optica (NMO)/NMO spectrum disorder (NMOSD). Similar to NMO/NMOSD associated with AQP4 antibodies, preceding infections have been reported in patients with MOG antibody-positive ON. This is the first report of bilateral ON following a herpes simplex virus (HSV) infection associated with a positive MOG antibody. Case presentation A 41-year-old man who initially presented with genital herpes developed allodynia in the Th2-Th5 and Th8-L2 areas, urinary retention, and painful visual loss in the left eye. Ophthalmological evaluation and brain magnetic resonance imaging (MRI) revealed bilateral ON. A spinal MRI showed leptomeningeal enhancement from the thoracic to lumbar vertebrae and abnormal enhancement of the L3 to S3 dorsal root ganglia without a change in intramedullary signals. Following treatment with acyclovir and steroid pulse, he fully recovered. Serum anti-AQP4 antibodies were negative, but anti-MOG antibodies were positive. Finally, he was diagnosed with MOG antibody-positive bilateral ON and meningoganglionitis following an HSV infection. Conclusion Our case supports a relationship between anti-MOG antibodies and ON triggered by an HSV infection. Clinicians should thus consider testing for MOG antibodies in patients with post-infectious neurological symptoms due to an HSV infection.

Summary Allografts of retinal pigment epithelial (RPE) cells have been considered for the treatment of ocular diseases. We recently started the transplantation of induced pluripotent stem cell (iPSC)-derived RPE cells for patients with age-related macular degeneration (autogenic grafts). However, there are at least two problems with this approach: (1) high cost, and (2) uselessness for acute patients. To resolve these issues, we established RPE cells from induced iPSCs in HLA homozygote donors. In vitro, human T cells directly recognized allogeneic iPSC-derived RPE cells that expressed HLA class I/II antigens. However, these T cells failed to respond to HLA-A, -B, and -DRB1-matched iPSC-derived RPE cells from HLA homozygous donors. Because of the lack of T cell response to iPSC-derived RPE cells from HLA homozygous donors, we can use these allogeneic iPSC-derived RPE cells in future clinical trials if the recipient and donor are HLA matched., Highlights • We established human RPE cells from iPSCs in HLA homozygote donors • iPS-RPE cells uniquely expressed HLA class I and class II molecules • T lymphocytes responded to allogeneic iPS-RPE cells, but not iPSCs, in vitro • T lymphocytes failed to respond to allogeneic iPS-RPE cells from HLA homozygote donors, In this article, Takahashi and colleagues show that the allogeneic immune response to iPSC-derived retinal pigment epithelial (RPE) cells by T cells in vitro. However, HLA-restricted immune reaction (at least HLA-A, -B, and -DRB1-matched) does not occur when iPS-RPE cells established from HLA homozygous donors are used.

Novel surface-modified, visible light-emitting and noncytotoxic ZnO nanoparticles (NPs) (ZPAZ) having aminotriethylene oxide chains linked by 1,4- and/or 1,5-disubstituted 1,2,3-triazole rings were prepared from ZnO NPs (ZPA) with ethynyl groups on the surfaces and an azide derivative of triethylene oxide chain linking terminal amino group (ATA) via 1,3-dipolar azide/alkyne click reaction by heating without Cu(I) catalyst. FTIR spectroscopy, elemental analysis, XRD analysis and TEM observation suggested that the resulting ZPA and ZPAZ NPs have the particle sizes below 10 nm in diameters, triethylene oxide chains linking the terminal amino groups and wurtzite crystal structure. UV–vis absorption spectrum of the ZPAZ NPs in methanol showed maximum absorption band at 346.5 nm, supporting the TEM observation. PL spectra depicted that the ZPA and ZPAZ NPs display broad light green and lightly greenish yellow visible light emitting bands in methanol. Zeta potentials measured in distilled water suggested that the ZPAZ NPs have a low tendency to aggregate and possess better stability than the ZPA NPs. Cytotoxicity assay revealed that the ZPAZ NPs, having water-dispersion properties, are noncytotoxic at low concentrations and almost all RAW264.7 cells are alive after 24 h of treatment.

A kinetic study of the quenching reaction of singlet oxygen (1O2) with eight kinds of carotenoids and α-tocopherol was performed in ethanol/chloroform/D2O (50:50:1, v/v/v) solution at 35 °C. The overall rate constants, kQ (=kq+kr, physical quenching+chemical reaction), for the reaction of carotenoids with 1O2 were measured, using the competition reaction method, where endoperoxide was used as a singlet oxygen generator, 2,5-diphenyl-3,4-benzofuran (DPBF) as an UV-vis absorption prove, and α-tocopherol as a standard compound. The rate constants, kQ (S) and kQ (t1/2), were determined by analyzing the first-order rate constant (S) and the half-life (t1/2) of the decay curve of DPBF with carotenoids, respectively, showing good accordance with each other. Similar measurements were performed for tomato and carrot extracts. From the results, a new assay method that can quantify the singlet oxygen absorption capacity (SOAC) of antioxidants, including carotenoids, α-tocopherol, and vegetable extracts, has been proposed.

Seasonal change in spermatogenesis was examined in the restricted spermatogonium-type testes of a teleost, Oryzias latipes. Histological observation revealed that the number of each stage of germ cells during most of the non-reproductive season, from October to January (O–J period) was nearly half of that during the reproductive season, from May to July (M–J period), except for type B spermatogonia (B-gonia), which was actually equal. As a result, the ratio of primary spermatocytes (P-cytes) to B-gonia was remarkably small in the O–J period. Despite the differences between both time periods, the proliferative activity of type A spermatogonia (A-gonia), B-gonia, or P-cytes was at a similar level in both periods. Moreover, in cultured testes treated with bromodeoxyuridine as a cell-lineage tracer, P-cytes differentiated to spermatids in 11–15 days in both M–J and O–J periods. These indicate that spermatogenesis is active in each period at a different state. In the spermatogenic testis, A-gonial proliferation was maintained by human follicle stimulating hormone/luteinizing hormone in culture. Whereas cell death of B-gonia and/or P-cytes gradually increased in the M–J period in spite of those cells being constant in population sizes. In transition to the O–J period, A-gonia and P-cytes first decreased, which was accompanied by a decrease in proliferative activity of A-gonia and relative increase of dead cells from B-gonia and/or P-cytes against live P-cytes. These suggest that A-gonial proliferation and cell death of B-gonia and/or P-cytes that is induced coordinately with B-gonial differentiation are critical for the spermatogenic control.

To determine the role of cholesterol synthesis in pancreatic b-cells, a transgenic model of in vivo activation of sterol-regulatory element binding protein 2 (SREBP-2) spe- cifically in b-cells (TgRIP-SREBP-2) was developed and ana- lyzed. Expression of nuclear human SREBP-2 in b-cells resulted in severe diabetes as evidenced by greater than 5-fold elevations in glycohemoglobin compared with C57BL/6 controls. Diabetes in TgRIP-SREBP-2 mice was pri- marily due to defects in glucose- and potassium-stimulated insulin secretion as determined by glucose tolerance test. Isolated islets of TgSREBP-2 mice were fewer in number, smaller, deformed, and had decreased insulin content. SREBP-2-expressing islets also contained increased esteri- fied cholesterol and unchanged triglycerides with reduced ATP levels. Consistently, these islets exhibited elevated ex- pression of HMG-CoA synthase and reductase and LDL re- ceptor, with suppression of endogenous SREBPs. Genes involved in b-cell differentiation, such as PDX1 and BETA2, were suppressed, explaining loss of b-cell mass, whereas IRS2 expression was not affected. These phenotypes were dependent on the transgene expression. Taken together, these results indicate that activation of SREBP-2 in b-cells caused severe diabetes by loss of b-cell mass with accumula- tion of cholesterol, providing a new lipotoxic model and a potential link of disturbed cholesterol metabolism to impair- ment of b-cell function.—Ishikawa, M., Y. Iwasaki, S. Yatoh, T. Kato, S. Kumadaki, N. Inoue, T. Yamamoto, T. Matsuzaka, Y. Nakagawa, N. Yahagi, K. Kobayashi, A. Takahashi, N. Yamada, and H. Shimano. Cholesterol accumulation and diabetes in pancreatic b-cell-specific SREBP-2 transgenic mice: a new model for lipotoxicity. J. Lipid Res. 2008. 49: 2524-2534.

Grixazone (GX), which is a diffusible yellow pigment containing a phenoxazinone chromophore, is one of the secondary metabolites under the control of A-factor (2-isocapryloyl-3 R -hydroxymethyl-γ-butyrolactone) in Streptomyces griseus . GX production is also induced by phosphate starvation. The whole biosynthesis gene cluster for GX was cloned and characterized. The gene cluster consisting of 13 genes contained six transcriptional units, griT , griSR , griR , griAB , griCDEFG , and griJIH . During cultivation in a phosphate-depleted medium, the six promoters were activated in the order (i) griR , (ii) griC and griJ , and (iii) griT , griS , and griA . Disruption of griR , which encodes a SARP family transcriptional regulator, abolished the transcriptional activation of all other genes in the cluster. In addition, ectopic expression of griR from a constitutively active promoter resulted in GX overproduction even in the absence of AdpA, a key transcriptional activator in the A-factor regulatory cascade, and in the presence of phosphate at a high concentration. GriR monomers bound direct repeat sequences in the griC and griJ promoters in a cooperative manner. Therefore, the early active genes ( griCDEFG and griJIH ), all of which, except for griG (which encodes a transporter-like protein), encode the GX biosynthesis enzymes, were directly activated by GriR. The transcription of griR was greatly reduced in the presence of phosphate at a high concentration and was hardly detected in the absence of AdpA. These findings showed that both A-factor and phosphate depletion signals were required for griR transcription and both signals were transmitted to the GX biosynthesis genes solely via the griR promoter.

Measurements of the second-order rate constants and the singlet oxygen absorption capacity (SOAC) values for the reaction of singlet oxygen (1O2) with 23 kinds of food extracts were performed in ethanol/chloroform/D2O (50:50:1, v/v/v) solution at 35 °C. It has been clarified that the SOAC method is useful to evaluate the 1O2-quenching activity (i.e. the SOAC value) of food extracts having two orders of magnitude different rate constants from 3.18 × 104 L g−1 s−1 for tomato to 1.55 × 102 for green melon. Furthermore, comparison of the observed rate constants for the above food extracts with the calculated ones based on the concentrations of seven kinds of carotenoids included in the food extracts and the rate constants reported for each carotenoids was performed, in order to ascertain the validity of the SOAC assay method developed and to clarify the ratio of the contribution of principal carotenoids to the SOAC value.

Exercise hypotension has been documented in patients with hypertrophic cardiomyopathy (HCM). Some investigators reported that this is due to an inadequate increase of systemic vascular resistance during or after exercise. The aim of this study was to investigate the pathogenesis of limb vascular response abnormalities in patients with idiopathic HCM. Thirteen patients with HCM and 9 healthy controls were administered an endothelium-dependent dilator (acetylcholine), a direct smooth muscle dilator (nitroglycerin), an alpha-adrenergic stimulator (phenylephrine), and a beta-adrenergic stimulator (isoproterenol) by intra-arterial infusion. Peripheral vascular resistance was assessed by forearm plethysmography at rest and during each infusion to obtain the percentage change in vascular resistance. Isoproterenol, acetylcholine and nitroglycerin produced dose-related vasodilatation with no significant differences between HCM patients and healthy controls. The percentage increase in vascular resistance was significantly less in HCM patients than in healthy controls after phenylephrine infusions of 0.5μg/min (155±38% vs 195±47%, mean ± SD, p

Background and hypothesis: Patients with dilated cardiomyopathy (DCM) with left ventricular hypertrophy (LVH) have been found to have a better prognosis than patients without LVH. However, the pathophysiologic mechanism for that has not been investigated. We sought to clarify the pathophysiologic significance of LVH in DCM. Methods: We performed isoproterenol infusion echocar-diography (0.02 mg/kg/min) in 17 patients with DCM, and measured plasma epinephrine and norepinephrine levels at rest and at the end of ergometer exercise in 14 of the 17 patients. Patients were classified into groups according to the presence (9 patients) (LVH+) or absence (8 patients) (LVH-) of LVH. Left ventricular hypertrophy was defined as an inter-ventricular thickness or posterior wall thickness ≥13 mm. Results: Although there was no significant difference between groups in fractional shortening at rest during isoproterenol infusion, fractional shortening was significantly higher in the LVH (+) group than in the LVH (-) group (29 ± 9 vs. 17 ± 8%;p

To determine whether there is a significant association between human immunodeficiency virus (HIV)-related retinal microangiopathy and systemic cytomegalovirus (CMV) disease in HIV-infected patients. Participants in this single-center, cross-sectional, retrospective study were 383 HIV-infected patients assessed for ocular manifestations before the beginning of antiretroviral therapy. The presence of HIV-related retinal microangiopathy, the presence of systemic CMV disease, laboratory data, and demographic information were determined by referring to medical records. The significance of any association between HIV-related retinal microangiopathy and systemic CMV disease was determined by use of the Chi-squared test and by multivariate analysis. HIV-related retinal microangiopathy was present in 85 patients, and was significantly associated with systemic CMV disease both by use of the Chi-squared test (P = 0.006) and by multivariate analysis (P = 0.045, odds ratio 2.03, 95 % confidence interval 1.02–4.06 adjusted for CD4+ cell count and plasma HIV-RNA level). These findings indicate that microangiopathy may be involved in the development of CMV disease in HIV-infected patients. Thus, detection of the presence of HIV-related retinal microangiopathy is important in the management of HIV-infected patients.

Objectives.We sought to assess whether isoproterenol stress echocardiography could detect in advance in which patients hypertrophic cardiomyopathy would progress to a phase resembling dilated cardiomyopathy.Background.In a few patients, hypertrophic cardiomyopathy has been reported to progress to a phase characterized by systolic dysfunction and left ventricular dilation, resembling dilated cardiomyopathy.Methods.Echocardiograms were recorded before and immediately after intravenous infusion of isoproterenol (0.02 μg/kg body weight per min) for 5 min in 18 patients with typical hypertrophic cardiomyopathy (i.e., hypertrophied, hyperdynamic and nondilated) to determine the difference in fractional shortening. The patients were categorized into those with a good response (difference in fractional shortening >7%, 14 patients) and those with a poor response (difference ≤ 7%, 4 patients). Changes in left ventricular end-diastolic diameter and fractional shortening were evaluated by using serial echocardiography over an average follow-up period of 5.4 years.Results.In the good response group, neither end-diastolic diameter nor fractional shortening changed significantly during the follow-up period. In the poor response group, end-diastolic diameter significantly increased from a mean ± SD of 41 ± 5 to 53 ± 5 mm (p < 0.05), and fractional shortening significantly decreased from 40 ± 12% to 29 ± 10% (p < 0.05). All patients in the poor response group showed a substantial decrease (≥ 5%) in fractional shortening and an increase (≥5 mm) in end-diastolic diameter. One patient developed congestive heart failure due to systolic dysfunction during the observation period.Conclusions.The present study confirmed that impaired responses to isoproterenol infusion are related to future deterioration of left ventricular performance in patients with typical hypertrophic cardiomyopathy.

Recently a new assay method that can quantify the singlet oxygen absorption capacity (SOAC) of antioxidants was proposed. In the present work, kinetic study of the reaction of singlet oxygen ((1)O(2)) with carotenoids and vegetable extracts has been performed in ethanol/chloroform/D(2)O (50:50:1, v/v/v) solution at 35 °C. Measurements of the second-order rate constants (k(Q)(S)) and the SOAC values were performed for eight kinds of carotenoids and three kinds of vegetable extracts (red paprika, carrot, and tomato). Furthermore, measurements of the concentrations of the carotenoids included in vegetable extracts were performed, using a HPLC technique. From the results, it has been clarified that the total (1)O(2)-quenching activity (that is, the SOAC value) for vegetable extracts may be explained as the sum of the product {Σ k(Q)(Car-i)(S) [Car-i](i)} of the rate constant (k(Q)(Car-i)(S)) and the concentration ([Car (i)]) of carotenoids included in vegetable extracts.

Seasonal change in spermatogenesis was examined in the restricted spermatogonium-type testes of a teleost, Oryzias latipes. Histological observation revealed that the number of each stage of germ cells during most of the non-reproductive season, from October to January (O-J period) was nearly half of that during the reproductive season, from May to July (M-J period), except for type B spermatogonia (B-gonia), which was actually equal. As a result, the ratio of primary spermatocytes (P-cytes) to B-gonia was remarkably small in the O-J period. Despite the differences between both time periods, the proliferative activity of type A spermatogonia (A-gonia), B-gonia, or P-cytes was at a similar level in both periods. Moreover, in cultured testes treated with bromodeoxyuridine as a cell-lineage tracer, P-cytes differentiated to spermatids in 11-15 days in both M-J and O-J periods. These indicate that spermatogenesis is active in each period at a different state. In the spermatogenic testis, A-gonial proliferation was maintained by human follicle stimulating hormone/luteinizing hormone in culture. Whereas cell death of B-gonia and/or P-cytes gradually increased in the M-J period in spite of those cells being constant in population sizes. In transition to the O-J period, A-gonia and P-cytes first decreased, which was accompanied by a decrease in proliferative activity of A-gonia and relative increase of dead cells from B-gonia and/or P-cytes against live P-cytes. These suggest that A-gonial proliferation and cell death of B-gonia and/or P-cytes that is induced coordinately with B-gonial differentiation are critical for the spermatogenic control.

Vietnam has been advancing toward a market economy since 1986. Industrialization has progressed with a high rate of growth. One of the factors of the economic growth of Vietnam has been FDI. Japanese companies are among those that have a strong interest in Vietnam. Japanese companies are recently taking note of Vietnam’s IT and software industries. Now, however, interest is increasing in offshoring as a means for developing in this sector.

Transgenic mice expressing nuclear sterol regulatory element-binding protein-1a under the control of the insulin promoter were generated to determine the role of SREBP-1a in pancreatic beta-cells. Only low expressors could be established, which exhibited mild hyperglycemia, impaired glucose tolerance, and reduced plasma insulin levels compared to C57BL/6 controls. The islets isolated from the transgenic mice were fewer and smaller, and had decreased insulin content and unaltered glucagon staining. Both glucose- and potassium-stimulated insulin secretions were decreased. The transgenic islets consistently expressed genes for fatty acids and cholesterol synthesis, resulting in accumulation of triglycerides but not cholesterol. PDX-1, BetaEpsilonTauAlpha2, MafA, and IRS-2 were suppressed, partially explaining the loss and dysfunction of beta-cell mass. The transgenic mice on a high fat/high sucrose diet still exhibited impaired insulin secretion and continuous beta-cell growth defect. Therefore, nuclear SREBP-1a, even at a low level, strongly disrupts beta-cell mass and function.

Insulin resistance is often associated with obesity and can precipitate type 2 diabetes. To date, most known approaches that improve insulin resistance must be preceded by the amelioration of obesity and hepatosteatosis. Here, we show that this provision is not mandatory; insulin resistance and hyperglycemia are improved by the modification of hepatic fatty acid composition, even in the presence of persistent obesity and hepatosteatosis. Mice deficient for Elovl6, the gene encoding the elongase that catalyzes the conversion of palmitate to stearate, were generated and shown to become obese and develop hepatosteatosis when fed a high-fat diet or mated to leptin-deficient ob/ob mice. However, they showed marked protection from hyperinsulinemia, hyperglycemia and hyperleptinemia. Amelioration of insulin resistance was associated with restoration of hepatic insulin receptor substrate-2 and suppression of hepatic protein kinase C epsilon activity resulting in restoration of Akt phosphorylation. Collectively, these data show that hepatic fatty acid composition is a new determinant for insulin sensitivity that acts independently of cellular energy balance and stress. Inhibition of this elongase could be a new therapeutic approach for ameliorating insulin resistance, diabetes and cardiovascular risks, even in the presence of a continuing state of obesity.

Well-preserved natural killer (NK) cell cytotoxicity (NKCC) is associated with healthy aging. The objective of the survey was to investigate psychological factors related to NKCC and NK cell populations in elderly women. A cross-sectional study involving 181 participants was conducted using the Japanese version of the 28-item General Health Questionnaire (GHQ) and additional questions assessing psychological status and lifestyle. Spearman's rank test revealed a significant negative correlation between NKCC and the GHQ depression subscale (GHQ-D) scores. Significantly reduced NKCC was found in participants presenting high GHQ-D scores (12or = GHQ-D, n = 58) compared with those showing middle (8or = GHQ-Dor = 11, n = 55) or low (GHQ-D = 7, n = 68) scores. Adjusting for covariates regarding lifestyle, multiple logistic regression analysis was applied; consequently, significant associations were found between reduced NKCC and high depressive symptoms and between increased NK cell numbers and life satisfaction. These results indicated a clue to longitudinal studies in the future.