Differentiation of natural killer cells from induced pluripotent stem cells under defined, serum- and feeder-free conditions.

BACKGROUND AIMS: Traditionally, natural killer (NK) cells are sourced from the peripheral blood of donors-a laborious and highly donor-specific process. Processes for generating NK cells from induced pluripotent stem cells (iPSCs) have demonstrated that it is possible to successfully generate renewable alloreactive NK cells that are not only functional in vivo but can also be genetically engineered for enhanced function. However, poor standardization and cumbersome differentiation procedures suggest that further improvements in the control of the differentiation process are necessary. METHODS: Here the authors evaluated the potential of differentiating NK cells from centrally authenticated iPSCs under entirely chemically defined and serum-free conditions as well as their immunotherapeutic potential, after expansion in feeder-free media, against solid tumors targets. To address limitations of current differentiation approaches, the authors did not utilize feeder or stromal cell layers, TrypLE adaptation or peripheral blood during the differentiation process. The authors also evaluated the feasibility of utilizing centrally authenticated iPSC lines, thus circumventing protocol- and donor-induced variability associated with reprogramming approaches, and characterized these iPSC-NK cells in terms of cytotoxicity, cytokine production and degranulation potential against solid tumor cell lines and patient-derived targets. RESULTS: Differentiation of iPSCs generated NK cells that were predominantly CD56+/CD16+/CD3- and expressed NK activation markers NKG2D, NKp30, NKp44, NKp46 and DNAM-1. These iPSC-NK cells mediated effector functions, including cytotoxicity, degranulation and IFN-γ production, in response to solid tumor targets, including patient-derived cancer cells, and could be cryopreserved and expanded in culture. CONCLUSIONS: The ability to produce NK cells under defined conditions and the functional responses elicited by these iPSC-NK cells suggest that they could represent promising effectors in clinical adoptive transfer settings as a renewable source of donor-independent NK cells for immunotherapy of solid tumors.

Differential impact of prostaglandin analogues on agreement of intraocular pressure measurements obtained by Goldmann applanation, rebound, and noncontact tonometry.

BACKGROUND: To evaluate the effect of topical prostaglandin analogues on agreement of IOP measurements obtained by Goldmann applanation tonometry (GAT), rebound tonometry (RBT), and noncontact tonometry (NCT) in eyes with primary open- angle glaucoma (POAG). METHODS: Intraocular pressure measurements were obtained using GAT, RBT, and NCT in patients with POAG with or without prostaglandin analogues. The agreement between each tonometry was analysed using Bland-Altman analyses in those with or without prostaglandin analogues. The effect of average IOP on IOP differences was also evaluated. RESULTS: Among a total of 86 subjects included in the study, 44 patients were using prostaglandin analogues. The difference in IOP measured by GAT and RBT was marginally greater in those with (GAT-RBT: - 0.94 ± 1.63 mmHg) prostaglandin analogues than in those without (- 0.33 ± 1.22 mmHg, P = 0.06). The difference in IOP measured by GAT and NCT was significantly greater in the prostaglandin group (GAT-NCT: 2.40 ± 2.89 mmHg) than in the group without prostaglandin analogues (0.41 ± 1.63 mmHg, P < 0.01). While there was no significant relationship between the average of all tonometries and the difference between tonometries in those without prostaglandin analogues, both RBT and NCT underestimated IOP relative to GAT at higher IOP in those using prostaglandin analogues. CONCLUSION: Intraocular pressure measured by RBT and NCT was similar to that measured by GAT in those without prostaglandin analogues. RBT overestimated and NCT underestimated IOP compared to GAT in those using prostaglandin analogues.

Antifibrotic effects of pirfenidone on Tenon's fibroblasts in glaucomatous eyes: comparison with mitomycin C and 5-fluorouracil.

PURPOSE: The purpose of this study was to evaluate the antifibrotic effects of pirfenidone (PFD) on primary cultured human Tenon's fibroblasts (HTFs) from primary open-angle glaucoma (POAG) eyes, compared to mitomicin C (MMC) and 5-fluorouracil (5-FU). MATERIALS AND METHODS: Samples of human Tenon's capsule were obtained during respective surgeries from three groups of patients: patients with cataract (CAT group), patients with POAG who underwent glaucoma filtration surgery (GFS) (POAG1 group), and patients with POAG who underwent GFS due to failed bleb of previous GFS (POAG2 group). Cell toxicity, cell migration, and the expression level of α-smooth muscle actin (α-SMA) protein were evaluated in primary cultured HTFs from the three patient groups after treatment (PFD, MMC, or 5-FU). RESULTS: Overall, cell viability after PFD treatment was higher compared to MMC treatment (82.3 ± 5.1 % vs 56.7 ± 3.8 %; p = 0.001) and comparable to 5-FU treatment (82.3 ± 5.1 % vs 85.7 ± 10.7 %, p = 0.214) at the same concentration (0.4 mg/ml). Both 0.3 mg/ml PFD and 0.1 mg/ml MMC inhibited cell migration compared to control (without treatment) cells (p = 0.014 and 0.005, respectively), while 0.2 mg/ml 5-FU showed the highest degree of cell migration among the three agents in the POAG1 group (PFD vs MMC vs 5-FU; 29.5 ± 2.1 % vs 34.5 ± 0.7 % vs 76.0 ± 8.5 %, PFD vs MMC; p = 1.000, PFD vs 5-FU; p = 0.008, MMC vs 5-FU; p = 0.011). PFD (0.1 or 0.3 mg/ml) and MMC (0.05 and 0.1 mg/ml) treatment significantly reduced the protein expression level of α-SMA in the POAG 1 group (all p < 0.05), and the α-SMA protein level following treatment with 0.3 mg/ml PFD was lower than that of 0.1 mg/ml MMC (p = 0.040). CONCLUSION: PFD showed less cytotoxicity compared to MMC. PFD and MMC inhibited cell migration and reduced α-SMA protein expression levels, while 5-FU showed neither inhibition of cell migration nor reduction in α-SMA expression level. These findings indicate PFD as a potential adjunctive antifibrotic agent to prevent bleb failure during GFS.

ES cell-derived renewable and functional midbrain dopaminergic progenitors.

During early development, midbrain dopaminergic (mDA) neuronal progenitors (NPs) arise from the ventral mesencephalic area by the combined actions of secreted factors and their downstream transcription factors. These mDA NPs proliferate, migrate to their final destinations, and develop into mature mDA neurons in the substantia nigra and the ventral tegmental area. Here, we show that such authentic mDA NPs can be efficiently isolated from differentiated ES cells (ESCs) using a FACS method combining two markers, Otx2 and Corin. Purified Otx2(+)Corin(+) cells coexpressed other mDA NP markers, including FoxA2, Lmx1b, and Glast. Using optimized culture conditions, these mDA NPs continuously proliferated up to 4 wk with almost 1,000-fold expansion without significant changes in their phenotype. Furthermore, upon differentiation, Otx2(+)Corin(+) cells efficiently generated mDA neurons, as evidenced by coexpression of mDA neuronal markers (e.g., TH, Pitx3, Nurr1, and Lmx1b) and physiological functions (e.g., efficient DA secretion and uptake). Notably, these mDA NPs differentiated into a relatively homogenous DA population with few serotonergic neurons. When transplanted into PD model animals, aphakia mice, and 6-OHDA-lesioned rats, mDA NPs differentiated into mDA neurons in vivo and generated well-integrated DA grafts, resulting in significant improvement in motor dysfunctions without tumor formation. Furthermore, grafted Otx2(+)Corin(+) cells exhibited significant migratory function in the host striatum, reaching >3.3 mm length in the entire striatum. We propose that functional and expandable mDA NPs can be efficiently isolated by this unique strategy and will serve as useful tools in regenerative medicine, bioassay, and drug screening.

The risk of open angle glaucoma in young adults with allergic diseases: a Nationwide cohort study.

This study investigated the potential associations between allergic diseases (asthma, allergic rhinitis, and atopic dermatitis) and the development of primary open-angle glaucoma. We utilized authorized data from the Korean National Health Information Database (KNHID), which provides comprehensive medical claims data and information from the National Health Screening Program. We compared the baseline characteristics of subjects with and without allergic diseases and calculated the incidence and risk of glaucoma development. Cox proportional hazard regression analysis was used to determine the risk of glaucoma development in subjects with allergic diseases. A total of 171,129 subjects aged 20-39 with or without allergic diseases who underwent a general health examination between 2009 and 2015 were included. Subjects with allergic diseases exhibited a higher incidence of glaucoma compared to the control group. The hazard ratio (HR) of glaucoma onset was 1.49 and 1.39 in subjects with at least one allergic disease before and after adjusting for potential confounding factors, respectively. Among allergic diseases, atopic dermatitis showed the highest risk for glaucoma development (aHR 1.73) after adjusting for confounders. Allergic rhinitis showed an increased risk for incident glaucoma after adjustment (aHR 1.38). Asthma showed the lowest but still increased risk for glaucoma (aHR 1.22). The associations were consistent in all subgroup analyses stratified by sex, smoking, drinking, exercise, diabetes, hypertension, dyslipidemia, or history of steroid. In conclusion, allergic diseases are associated with increased risk of glaucoma development. Among allergic diseases, atopic dermatitis showed the highest risk for glaucoma development followed by allergic rhinitis and asthma.

Presence and severity of migraine is associated with development of primary open angle glaucoma: A population-based longitudinal cohort study.

PURPOSE: To examine the association between the presence and severity of migraine and development of primary open-angle glaucoma (POAG) using a nationwide population-based longitudinal cohort data. METHODS: Data were retrieved from the Korean National Health Insurance Service for 2,716,562 individuals aged ≥ 40 years and assessed for the development of POAG from 2009 through 2018. Subjects were classified into the following 3 groups: healthy control subjects, subjects with mild migraine, and those with severe migraine. Hazard ratios (HR) of glaucoma development were calculated for each group. Subgroup analyses of subjects stratified by age, sex, lifestyle factors (smoking, drinking, and body mass index (BMI)), and comorbidities (diabetes, hypertension, and dyslipidemia). RESULTS: During the 9-year follow-up period, the incidence rate of POAG per 1000 person-years was 2.41 and 3.25 in subjects without and with migraine, respectively. Among the migraine group, the incidence rate was 3.14 and 3.89 in mild and severe subgroups, respectively. The HR was 1.355 (95% CI, 1.300-1.412) and 1.188 (95% CI, 1.140-1.239) before and after adjusting for potential confounding factors in the migraine group per se. Regarding the severity of migraine, the adjusted HRs were 1.169 (95% CI, 1.117-1.224) in the mild migraine group, and 1.285 (95% CI, 1.166-1.415) in the severe migraine group compared to the control group. The results were consistent in subgroup analyses after stratifying by age, sex, lifestyle factors, and comorbidities. CONCLUSIONS: Migraine is associated with increased risk of POAG development. Furthermore, chronic and severe migraine is associated with greater risk of POAG development.

Association between localized retinal nerve fiber layer defects in nonglaucomatous eyes and metabolic syndrome: a propensity score-matched analysis.

Background: We investigated the association between metabolic syndrome and localized retinal nerve fiber layer (RNFL) defects in nonglaucomatous subjects. Methods: We examined 20,385 adults who visited the Health Promotion Center of Seoul St. Mary's Hospital between May 2015 and April 2016. After excluding those with known glaucoma or glaucomatous optic discs, subjects with and without localized RNFL defects were 1:5 propensity score matched. Metabolic syndrome components, including central obesity, elevated triglyceride, reduced high-density lipoprotein (HDL) cholesterol, elevated blood pressure (BP), and elevated fasting glucose, were compared between two groups. We performed logistic regression to investigate the association between RNFL defects and each component of metabolic syndrome and the number of metabolic syndrome components. Results: Subjects with RNFL defects showed higher waist-to-hip ratios, systolic BP (SBP) and diastolic BP (DBP), fasting blood glucose, and hemoglobin A1c (HbA1c) levels than did those without RNFL defects both before and after propensity score matching. The number of metabolic syndrome components was significantly greater in those with RNFL defects (1.66±1.35) than in those without (1.27±1.32, P<0.01). In multivariate logistic regression, the odds ratio (OR) of RNFL defects was significantly increased in subjects with central obesity [OR =1.53, 95% confidence interval (CI): 1.11-2.13], elevated BP (OR =1.50, 95% CI: 1.09-2.05), and an elevated fasting glucose level (OR =1.42, 95% CI: 1.03-1.97). An increased number of metabolic syndrome components was associated with a higher risk of RNFL defects. Conclusions: Localized RNFL defects in nonglaucomatous subjects are associated with metabolic syndrome components, including central obesity, elevated BP, and an elevated fasting glucose level, suggesting that comorbid metabolic syndrome should be considered when evaluating subjects with RNFL defects.

Metabolic Health, Obesity, and Intraocular Pressure.

Obesity has been associated with increased intraocular pressure (IOP), but the results are inconsistent. Recently, a subgroup of obese individuals with good metabolic profiles were suggested to have better clinical outcomes than normal-weight individuals with metabolic diseases. The relationships between IOP and different combinations of obesity and metabolic health status have not been investigated. Therefore, we investigated the IOP among groups with different combinations of obesity status and metabolic health status. We examined 20,385 adults aged 19 to 85 years at the Health Promotion Center of Seoul St. Mary's Hospital between May 2015 and April 2016. Individuals were categorized into four groups according to obesity (body mass index (BMI) ≥ 25 kg/m2) and metabolic health status (defined based on prior medical history or abdominal obesity, dyslipidemia, low high-density lipoprotein cholesterol, high blood pressure, or high fasting blood glucose levels upon medical examination). ANOVA and ANCOVA were performed to compare the IOP among the subgroups. The IOP of the metabolically unhealthy obese group (14.38 ± 0.06 mmHg) was the highest, followed by that of the metabolically unhealthy normal-weight group (MUNW, 14.22 ± 0.08 mmHg), then, the metabolically healthy groups (p < 0.001; 13.50 ± 0.05 mmHg and 13.06 ± 0.03 mmHg in the metabolically healthy obese (MHO) and metabolically healthy normal-weight groups, respectively). Subjects who were metabolically unhealthy showed higher IOP compared to their counterparts who were metabolically healthy at all BMI levels, and there was a linear increase in IOP as the number of metabolic disease components increased, but no difference between normal-weight vs. obese individuals. While obesity, metabolic health status, and each component of metabolic disease were associated with higher IOP, those who were MUNW showed higher IOP than those who were MHO, which indicates that metabolic status has a greater impact than obesity on IOP.

Effects of AFP-172 on COX-2-induced angiogenic activities on human umbilical vein endothelial cells.

PURPOSE: To investigate the angiogenic effect of the free acid of tafluprost (AFP-172) on human umbilical vascular endothelial cells (HUVECs). METHODS: HUVECs cultured in the presence or absence of FP receptor antagonist (10 nM AL-8810) were exposed to escalating concentrations of 10(-7), 10(-6), 10(-5), 10(-4) and 10(-3) M AFP-172 (the free acid of tafluprost). For cell proliferation assays, the numbers of cells were derived from a CellTiter96® Aqueous One Solution Cell Proliferation Assay (Promega) by Microplate reader (Bio-Rad, Benchmark). Endothelial cell migration was evaluated by a BD Biocoat™ Angiogenesis System using FluoroBlok ™ 24-well inserts (BD Biosciences, Bedford, MA). BioTek FLx800 fluorescence plate reader was used for quantitative measurement of fluorescently-labeled invasive vascular endothelial cells. Endothelial capillary-like tube formation was evaluated by BD Biocoat Angiogenesis System using Matrigel Matrix 96-well plate. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assess the gene expression of vascular endothelial growth factor (VEGF), cyclooxygenase-2 (COX-2) and endothelial nitric oxide synthase (eNOS). COX-2 protein was detected by immunofluorescent staining and Western blot assay. Student's t-test was used for statistical analysis. RESULTS: 10(-4) M AFP-172 treated cells stimulated the proliferation, migration and tube formation of HUVECs as compared to 10(-5), 10(-6,) 10(-7) M AFP-172 treated cells and control (P < 0.01). RT-PCR showed that incubation of HUVECs with 10(-4) M AFP-172 stimulated the expression of COX-2 mRNA (P < 0.05). Western blot assay revealed that AFP-172 caused cells to increase in COX-2 protein at the concentrations of 10(-4) M. CONCLUSIONS: >AFP-172 showed the angiogenic effects on HUVECs at the concentrations of 10(-4) M by inducing COX-2 protein.

The effect of additional topical cyclosporine or vitamin A on the ocular surface during antiglaucoma medication administration.

PURPOSE: To investigate the effects of topical application of cyclosporine or vitamin A on the ocular surface during the concurrent administration of antiglaucoma drugs. METHODS: Thirty rabbits were randomized into 5 groups. Group 1 was administered timolol, group 2 received travoprost, group 3 received a travoprost/timolol fixed combination solution, group 4 received timolol and travoprost, and group 5 received timolol, travoprost, and dorzolamide. Each group was divided into a subgroup that received only the antiglaucoma medication (subgroup A), a subgroup that received topical cyclosporine in addition to the antiglaucoma medication (subgroup B), and a subgroup that received topical vitamin A in addition to the antiglaucoma medication (subgroup C). Conjunctival impression cytology specimens were collected at baseline and at weeks 1, 3, and 6. Conjunctival biopsy specimens were collected at week 6. RESULTS: The impression cytologic study results are as follows: statistically significant differences were found between groups 4A and 4B and between groups 4A and 4C at week 6 (p = 0.004, p = 0.006, respectively) and between groups 5A and 5B and between groups 5A and 5C at weeks 3 and 6 (p = 0.006, p = 0.008 at week 3, p = 0.003, p = 0.004 at week 6, respectively). No statistically significant differences were found between subgroup B and subgroup C in any of the groups at any of the times evaluated (p > 0.05). The conjunctival biopsy specimens from groups 1, 2, and 3 showed no distortion, but groups 4A and 5A showed distortion of the conjunctival epithelial structures. Groups 4B, 4C, 5B, and 5C showed less distortion of the conjunctival epithelial structures. CONCLUSION: Administration of cyclosporine or vitamin A may reduce the adverse ocular surface changes caused by long-term administration of antiglaucoma drugs.

Measurement of corneal biomechanical properties in diabetes mellitus using the Corvis ST.

We sought to assess changes in corneal biomechanical parameters in patients with diabetes mellitus (DM) in comparison with those among healthy controls using Corvis ST (CST). The study group included 209 eyes from healthy control subjects and 33 eyes from diabetic subjects, respectively. Following an ophthalmological examination, measurements with CST were taken. Additionally, hemoglobin A1c and blood glucose values were collected. Results were then compared to those of the control group after adjusting for potential confounding factors, including age-, intraocular pressure (IOP)-, central corneal thickness (CCT)-, spherical equivalent (SE)- and axial length (AL). After adjusting for potential confounding factors, including the age, IOP, CCT, SE, and AL, patients with DM presented significantly lower whole-eye movement (WEM) (ms) values than patients without DM (21.71 ± 0.84 vs. 22.15 ± 0.64 ms; P < .001). There was a significant and negative correlation between WEM (ms) and hemoglobin A1c in DM patients (r = -0.733; P = .001). In univariate and multivariate general linear mixed model (GLMM) analyses, IOP (P < .001 and P < .001, respectively) and the presence of DM (P = .001 and P < .001, respectively) significantly affected WEM (ms). In DM, significant changes in corneal biomechanical properties were detectable. The DM group showed significantly less deformable cornea and sclera than did the normal controls, even after adjusting for age, IOP, CCT, SE, and AL. These findings may cause misinterpretation of IOP measurements in diabetic patients. Therefore, the measurement of corneal biomechanics should be taken into consideration in clinical practice.

Impact of vision and hearing impairments on risk of cardiovascular outcomes and mortality in patients with type 2 diabetes: A nationwide cohort study.

AIMS/INTRODUCTION: The purpose of this study was to investigate the impact of vision and hearing impairments on the risk of adverse cardiovascular outcomes and mortality in patients with type 2 diabetes using a nationwide longitudinal cohort. MATERIALS AND METHODS: We enrolled 771,128 patients with type 2 diabetes who underwent the National Health Screening Program in 2009. We carried out Cox proportional hazards regression analyses to calculate the hazard ratios (HR) of myocardial infarction (MI), stroke, and mortality in those with or without vision and hearing impairments. Subgroup analyses of patients stratified by age, sex and diabetic retinopathy were carried out. RESULTS: Diabetes patients with either vision or hearing impairment showed higher risk of MI, stroke or death compared with those without. Among the combinations of impairments, patients with both vision and hearing impairments had the highest risk for MI (adjusted HR [aHR] 1.362, 95% confidence interval [CI] 1.252-1.481) and mortality (aHR 1.591, 95% CI 1.532-1.651). Those with only vision impairment showed higher risk of MI (aHR 1.324, 95% CI 1.275-1.375 and aHR 1.117, 95% CI 1.066-1.170, respectively), stroke (aHR 1.318, 95% CI 1.276-1.362 and aHR 1.134 95% CI 1.089-1.180, respectively) and mortality (aHR 1.417, 95% CI 1.390-1.446 and aHR 1.163, 95% CI 1.135-1.191, respectively) compared with those with only hearing impairment. CONCLUSIONS: Vision and hearing impairments are independently important risk factors for adverse cardiovascular events and mortality in patients with type 2 diabetes. Vision and hearing impairments synergistically increased the risk of MI and all-cause deaths, but not stroke. In addition, in patients aged <65 years, the HR of vision impairment was higher than those with vision and hearing impairments.

Effect of depressive symptom and depressive disorder on glaucoma incidence in elderly.

Although depression and glaucoma share several common pathophysiology, the risk of glaucoma in patients with depression has not been reported. Thus, we investigated the effect of depressive symptom and depressive disorder on glaucoma incidence. In this nationwide population-based cohort study, all subjects receiving the National Screening Program at the age of 66 during 2009-2014 were included. These subjects were divided into depression group and no depression group based on subjective depressive symptoms and clinically diagnosed depressive disorder and were tracked until 2017 for development of glaucoma. Of the 922,769 subjects included in the study, 191,636 (20.77%) subjects were categorized as depression group. Subjects with depression showed increased hazard of developing glaucoma (adjusted HR = 1.12[95% confidence interval (CI), 1.09-1.15]) than those without depression. The risk of glaucoma increased sequentially from those with no depression to those with subjective depressive symptom (adjusted HR = 1.09[95% CI, 1.06-1.13]), those with clinically diagnosed depressive disorder (adjusted HR = 1.23[95% CI, 1.14-1.32]), and those with both subjective depressive symptom and clinically diagnosed depressive disorder (adjusted HR = 1.36[95% CI, 1.22-1.52]). Our analyses suggest that individuals with depression had a greater risk of developing glaucoma than those without depression. Subjective depressive symptoms and clinically diagnosed depressive disorder independently and synergistically increased the risk of glaucoma incidence.

Clinically compatible advances in blood-derived endothelial progenitor cell isolation and reprogramming for translational applications.

The promise of using induced pluripotent stem cells (iPSCs) for cellular therapies has been hampered by the lack of easily isolatable and well characterized source cells whose genomes have undergone minimal changes during their processing. Blood-derived late-outgrowth endothelial progenitor cells (EPCs) are used for disease modeling and have potential therapeutic uses including cell transplantation and the translation of induced pluripotent stem cell (iPSC) derivatives. However, the current isolation of EPCs has been inconsistent and requires at least 40-80 mL of blood, limiting their wider use. In addition, previous EPC reprogramming methods precluded the translation of EPC-derived iPSCs to the clinic. Here a series of clinically-compatible advances in the isolation and reprogramming of EPCs is presented, including a reduction of blood sampling volumes to 10 mL and use of highly efficient RNA-based reprogramming methods together with autologous human serum, resulting in clinically relevant iPSCs carrying minimal copy number variations (CNVs) compared to their parent line.

Relation between grade II hypertensive retinopathy and coronary artery disease in treated essential hypertensives.

It is known that in advanced hypertensive retinopathy, which changes advanced hypertensive retinopathy (Grade III or IV), there is a strong relation between retinal microvascular lesions and cardiac and macrovascular markers of target organ damage (TOD). The prevalence of grade II hypertensive retinopathy and its relationship to cardiovascular risk factors remain controversial. The subjects, a total of 437 hypertensive patients, were divided into three groups according to modified Keith, Wagener, and Barker (KWB) classification by two ophthalmologists: Grade 0 with normal retinal change (N = 169, 38.7%), Grade I with arteriolar narrowing (N = 215, 49.1%), Grade II with arteriovenous crossings (N = 49, 11.2%). The prevalence of Grade I and Grade II hypertensive retinopathy was significantly higher than that of advanced hypertensive retinopathy. The grade of hypertensive retinopathy was related to age, duration of hypertension, coronary artery disease (CAD), and left ventricular hypertrophy (LVH). The prevalence of LVH and CAD in Grade II was significantly higher than in Grade I and Grade 0. The hypertensive retinopathy Grade II was significantly correlated with LVH (odds ratio (OR) 2.3, 95% confidence interval (CI) 1.21-4.44, p < 0.05) and CAD (OR 4.2, 95% CI 1.97-8.95, p-<-0.001). Grade I and Grade II hypertensive retinopathy are frequently observed in hypertensive patients compared to Grade III and IV patients. We concluded that Grade II hypertensive retinopathy is closely related to CAD and should therefore not be ignored.

Risk of cancer in patients with glaucoma: A nationwide population-based cohort study.

To compare the risk of cancer development between patients with glaucoma and those without, we conducted a nationwide population-based cohort study using the Korean National Health Insurance Database. Individuals with diagnosis of glaucoma between 2007 and 2016 were identified, and controls were 1:1 matched based on age and sex. We calculated the incidence rates(IR) and hazard ratios(HR) before and after adjusting for age, gender, diabetes, smoking history, and body mass index. A total of 107,536 individuals with glaucoma and the same number of individuals without glaucoma were included. The IR of overall cancer were 12.23 and 11.62 per 1,000 individuals in the glaucoma and control groups, respectively. The HR of overall cancer was significantly higher in the glaucoma group before(HR: 1.053) and after adjusting for confounding factors(adjusted HR: 1.049) compared to that in the control group. The risk of overall cancer and specific cancers varied depending on gender and age groups, and the association was stronger in women and those under 65 years of age. Our study revealed that individuals with glaucoma showed higher risk of overall cancer and higher risk of specific cancers than those without glaucoma.

Target-dependent inhibition of sympathetic neuron growth via modulation of a BMP signaling pathway.

Target-derived factors modulate many aspects of peripheral neuron development including neuronal growth, survival, and maturation. Less is known about how initial target contact regulates changes in gene expression associated with these developmental processes. One early consequence of contact between growing sympathetic neurons and their cardiac myocyte targets is the inhibition of neuronal outgrowth. Analysis of neuronal gene expression following this contact revealed coordinate regulation of a bone morphogenetic protein (BMP)-dependent growth pathway in which basic helix-loop-helix transcription factors and downstream neurofilament expression contribute to the growth dynamics of developing sympathetic neurons. BMP2 had dose-dependent growth-promoting effects on sympathetic neurons cultured in the absence, but not the presence, of myocyte targets, suggesting that target contact alters neuronal responses to BMP signaling. Target contact also induced the expression of matrix Gla protein (MGP), a regulator of BMP function in the vascular system. Increased MGP expression inhibited BMP-dependent neuronal growth and MGP expression increased in sympathetic neurons during the period of target contact in vivo. These experiments establish MGP as a novel regulator of BMP function in the nervous system, and define developmental transitions in BMP responses during sympathetic development.

Corneal deflection amplitude and visual field progression in primary open-angle glaucoma.

PURPOSE: To investigate the relationship between corneal deflection amplitude and visual field progression rate in patients with primary open-angle glaucoma (POAG). METHODS: This study included 113 eyes of 65 patients with POAG followed for an average of 4.81 ± 1.24 years. Evaluation of visual field progression rate was performed using mean deviation of standard automated perimetry. Corneal deflection amplitude was measured using Corvis ST (Oculus Optikgeräte GmbH, Wetzlar, Germany). Linear mixed models were performed to determine the relationship between corneal deflection amplitude, intraocular pressure (IOP), and visual field progression rate. RESULTS: Mean age was 56.36 ± 14.58 years. Baseline average mean deviation was -8.20 ± 9.12 dB and mean treated IOP was 14.38 ± 3.08 mmHg. Average deflection amplitude was 0.90 ± 0.13 mm. In both univariate and multivariate analysis, IOP (P = 0.028 and P < 0.001, respectively) and deflection amplitude (P = 0.034 and P < 0.001, respectively) significantly affected visual field progression rate. Eyes with high IOP and greater deflection amplitude showed faster progression rate. CONCLUSIONS: Corneal deflection amplitude was significantly related with glaucoma progression. Eyes with greater corneal deflection amplitude showed faster visual field progression rate in patients with POAG.

Enhanced delivery of protein fused to cell penetrating peptides to mammalian cells.

Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols [BMB Reports 2019; 52(5): 324-329].

Glutathione depletion induces differential apoptosis in cells of mouse retina, in vivo.

Oxidative stress affects numerous intracellular macromolecules, and may result in cell death unless precisely regulated. Unregulated oxidative stress can be controlled by various cellular defense mechanisms such as glutathione (GSH) which can critically counteract the damaging effects of oxidative stress in mammalian cells. We determined the effects of unregulated oxidative stress induced by GSH depletion on cells in mouse retina. Mice were intraperitoneally injected with buthionine sulphoximine (BSO) at 1.5 g/kg. After 0, 1, 4, and 7 days of BSO administration, retinas were excised and sections were subjected to GSH assay and terminal uridine deoxynucleotidyl nick end labeling (TUNEL) analysis. After 4 days of BSO administration, the number of TUNEL positive cells was significantly increased. However, after 7 days, TUNEL positive cells returned to the basal level. The retinal region most affected by the BSO treatment appeared to be the outer nuclear layer where the photoreceptor cells reside. Different from cells in other regions, retinal cells in the inner nuclear layer increased in their apoptosis even after the first day of BSO injection, and the increase was further potentiated after 4 days. Taken together, our studies suggested that GSH depletion may cause unregulated oxidative stress to the cells in the retina and indeed increased cell death in the retina. The cells in the inner nuclear layer seemed to be affected earlier than the cells in other layers of the retina. The GSH level in the retina may be a crucial therapeutic target in preventing blindness.