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Consumption of coffee is associated with reduced risk of Parkinson's disease (PD), an effect that has largely been attributed to caffeine. However, coffee contains numerous components that may also be neuroprotective. One of these compounds is eicosanoyl-5-hydroxytryptamide (EHT), which ameliorates the phenotype of α-synuclein transgenic mice associated with decreased protein aggregation and phosphorylation, improved neuronal integrity and reduced neuroinflammation. Here, we sought to investigate if EHT has an effect in the MPTP model of PD. Mice fed a diet containing EHT for four weeks exhibited dose-dependent preservation of nigral dopaminergic neurons following MPTP challenge compared to animals given control feed. Reductions in striatal dopamine and tyrosine hydroxylase content were also less pronounced with EHT treatment. The neuroinflammatory response to MPTP was markedly attenuated, and indices of oxidative stress and JNK activation were significantly prevented with EHT. In cultured primary microglia and astrocytes, EHT had a direct anti-inflammatory effect demonstrated by repression of lipopolysaccharide-induced NFκB activation, iNOS induction, and nitric oxide production. EHT also exhibited a robust anti-oxidant activity in vitro. Additionally, in SH-SY5Y cells, MPP(+)-induced demethylation of phosphoprotein phosphatase 2A (PP2A), the master regulator of the cellular phosphoregulatory network, and cytotoxicity were ameliorated by EHT. These findings indicate that the neuroprotective effect of EHT against MPTP is through several mechanisms including its anti-inflammatory and antioxidant activities as well as its ability to modulate the methylation and hence activity of PP2A. Our data, therefore, reveal a strong beneficial effect of a novel component of coffee in multiple endpoints relevant to PD. [ABSTRACT FROM AUTHOR]

This study investigated 2 hypotheses about genotype-phenotype relationships for the efflux transporter, P-glycoprotein: (1) the presence of a synonymous C3435T variant in exon 26 of the MDR1 gene correlates to higher plasma concentrations of a P-glycoprotein substrate, dicloxacillin, and (2) the effects of genotypic differences decrease under conditions of P-glycoprotein induction by rifampin. Eighteen healthy volunteers received two 1-g doses of dicloxacillin, one on the 1st study day and the other on the 11th day of rifampin dosing (600 mg daily). Dicloxacillin and its 5-hydroxymethyl metabolite were analyzed using liquid chromatography/tandem mass spectrometry. Mean dicloxacillin C(max) measurements were 30.5 +/- 13.5, 33.3 +/- 4.7, and 31.1 +/- 12.8 mug/mL in individuals with the CC, CT, and TT genotype at position 3435 in exon 26 of the MDR1 gene. Following rifampin dosing, the mean dicloxacillin C(max) across genotypes decreased from 31.4 +/- 10.8 to 22.9 +/- 7.0 microg/mL (P < .05), whereas the mean oral clearance increased from 235 +/- 82 to 297 +/- 71 mL/min (P < .001), and the mean absorption time increased from 0.71 +/- 0.55 to 1.34 +/- 0.77 h (P < .05). Rifampin treatment increased the formation clearance, C(max), and AUC of the 5-hydroxymethyl metabolite by 135%, 119%, and 59%, respectively. The C3435T variant had no effect on dicloxacillin pharmacokinetics. The data suggested that rifampin induced intestinal P-glycoprotein and increased dicloxacillin metabolism. [ABSTRACT FROM AUTHOR]

Neovascular age-related macular degeneration (nAMD) is characterized by abnormal blood vessel growth from the choroid, leading to complications and eventual blindness. Despite anti-VEGF therapy, subretinal fibrosis remains a major concern, as VEGF/VEGF receptor-2 (VEGFR2) signaling can contribute to both angiogenesis and fibrosis. For the identification of the aqueous humor proteome, we performed liquid chromatography with tandem mass spectrometry analysis. To investigate the potential therapeutic effects of targeting the VEGF signaling pathway using apatinib, a highly selective VEGFR2 tyrosine kinase inhibitor, this study employed in vitro (THP-1 conditioned media-treated ARPE-19 cells) and in vivo (laser-induced choroidal neovascularization mouse) models of nAMD. This study revealed elevated VEGFR2 protein levels in the aqueous humor of nAMD patients, suggesting a potential target to mitigate neovascularization and fibrosis in nAMD. Apatinib effectively reduced VEGFA and αSMA levels in both in vitro and in vivo models. Moreover, apatinib showed improvement in laser-induced subretinal hyper-reflective lesions. The action mechanism was linked to the inhibition of VEGFR2 activation, leading to the suppression of both angiogenesis and fibrosis through the downregulation of STAT3 phosphorylation. Therefore, the VEGFR2 signaling pathway appears to play a central role in the development of nAMD by regulating both angiogenesis and fibrosis., (© 2024. The Author(s).)

α,ω-Dicarboxylic acids, ω-aminoalkanoic acids, and α,ω-diaminoalkanes are valuable building blocks for the production of biopolyesters and biopolyamides. One of the key steps in producing these chemicals is the oxidation of ω-hydroxycarboxylic acids using alcohol dehydrogenases (e.g., ChnD of Acinetobacter sp. NCIMB 9871). However, the reaction and structural features of these enzymes remain mostly undiscovered. Thereby, we have investigated characteristics of ChnD based on enzyme kinetics, substrate-docking simulations, and mutation studies. Kinetic analysis revealed a distinct preference of ChnD for medium chain ω-hydroxycarboxylic acids, with the highest catalytic efficiency of 18.0 mM -1 s -1 for 12-hydroxydodecanoic acid among C6 to C12 ω-hydroxycarboxylic acids. The high catalytic efficiency was attributed to the positive interactions between the carboxyl group of the substrates and the guanidino group of two arginine residues (i.e., Arg62 and Arg266) in the substrate binding site. The ChnD&#95;R62L variant showed the increased efficiency and affinity, particularly for fatty alcohols (i.e., C6-C10) and branched-chain fatty alcohols, such as 3-methyl-2-buten-1-ol. Overall, this study contributes to the deeper understanding of medium-chain primary aliphatic alcohol dehydrogenases and their applications for the production of industrially relevant chemicals such as α,ω-dicarboxylic acids, ω-aminoalkanoic acids, and α,ω-diaminoalkanes from renewable biomass., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study was carried out to screen the antifungal activity against Colletotrichum acutatum, Colletotrichum dematium, and Colletotrichum coccodes. Bacterial isolate GP-P8 from pepper soil was found to be effective against the tested pathogens with an average inhibition rate of 70.7% in in vitro dual culture assays. 16S rRNA gene sequencing analysis result showed that the effective bacterial isolate as Bacillus siamensis. Biochemical characterization of GP-P8 was also performed. According to the results, protease and cellulose, siderophore production, phosphate solubilization, starch hydrolysis, and indole-3-acetic acid production were shown by the GP-P8. Using specific primers, genes involved in the production of antibiotics, such as iturin, fengycin, difficidin, bacilysin, bacillibactin, surfactin, macrolactin, and bacillaene were also detected in B. siamensis GP-P8. Identification and analysis of volatile organic compounds through solid phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS) revealed that acetoin and 2,3-butanediol were produced by isolate GP-P8. In vivo tests showed that GP-P8 significantly reduced the anthracnose disease caused by C. acutatum, and enhanced the growth of pepper plant. Reverse transcription polymerase chain reaction analysis of pepper fruits revealed that GP-P8 treated pepper plants showed increased expression of immune genes such as CaPR1, CaPR4, CaNPR1, CaMAPK4, CaJA2, and CaERF53. These results strongly suggest that GP-P8 could be a promising biocontrol agent against pepper anthracnose disease and possibly a pepper plant growth-promoting agent.

Background: Bone readiness for implant placement is typically evaluated by bone quality/density on 2-dimensional radiographs and cone beam computed tomography at an arbitrary time between 3 and 6 months after tooth extraction and alveolar ridge preservation (ARP). The aim of this study is to investigate if high-frequency ultrasound (US) can classify bone readiness in humans, using micro-CT as a reference standard to obtain bone mineral density (BMD) and bone volume fraction (BVTV) of healed sockets receiving ARP in humans., Methods: A total of 27 bone cores were harvested during the implant surgery from 24 patients who received prior extraction with ARP. US images were taken immediately before the implant surgery at a site co-registered with the tissue biopsy collection location, made possible with a specially designed guide, and then classified into 3 tiers using B-mode image criteria (1) favorable, (2) questionable, and (3) unfavorable. Bone mineral density (hydroxyapatite) and BVTV were obtained from micro-CT as the gold standard., Results: Hydroxyapatite and BVTV were evaluated within the projected US slice plane and thresholded to favorable (>2200 mg/cm 3 ; >0.45 mm 3 /mm 3 ), questionable (1500-2200 mg/cm 3 ; 0.4-0.45 mm 3 /mm 3 ), and unfavorable (<1500 mg/cm 3 ; <0.4 mm 3 /mm 3 ). The present US B-mode classification inversely scales with BMD. Regression analysis showed a significant relation between US classification and BMD as well as BVTV. T-test analysis demonstrated a significant correlation between US reader scores and the gold standard. When comparing Tier 1 with the combination of Tier 2 and 3, US achieved a significant group differentiation relative to mean BMD (p = 0.004, true positive 66.7%, false positive 0%, true negative 100%, false negative 33.3%, specificity 100%, sensitivity 66.7%, receiver operating characteristics area under the curve 0.86). Similar results were found between US-derived tiers and BVTV., Conclusion: Preliminary data suggest US could classify jawbone surface quality that correlates with BMD/BVTV and serve as the basis for future development of US-based socket healing evaluation after ARP., (© 2024 The Authors. Journal of Periodontology published by Wiley Periodicals LLC on behalf of American Academy of Periodontology.)

Introduction: The abnormal hyperreactivity to food cues in individuals with binge eating behaviors could be regulated by hedonic or reward-based system, overriding the homeostatic system. The aim of the present study was to investigate whether attentional bias for food cues is affected by the level of hunger, maintaining the normal homeostatic system in individuals with binge eating behaviors., Methods: A total of 116 female participants were recruited and divided into four groups: hungry-binge eating group (BE) ( n = 29), satiated BE ( n = 29), hungry-control ( n = 29), satiated control ( n = 29). While participants completed a free-viewing task on high or low-calorie food cues, visual attentional processes were recorded using an eye tracker., Results: The results revealed that BE group showed longer initial fixation duration toward high-calorie food cues in both hunger and satiety condition in the early stage, whereas the control group showed longer initial fixation duration toward high-calorie food cues only in hunger conditions. Moreover, in the late stage, the BE group stared more at the high-calorie food cue, compared to control group regardless of hunger and satiety., Discussion: The findings suggest that automatic attentional bias for food cues in individuals with binge eating behaviors occurred without purpose or awareness is not affected by the homeostatic system, while strategic attention is focused on high-calorie food. Therefore, the attentional processing of food cues in binge eating group is regulated by hedonic system rather than homeostatic system, leading to vulnerability to binge eating., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Woo, Lee and Lee.)

A slot structure was combined with a discharge electrode to limit incident high-power microwaves via the integration of plasma discharge. At the target resonating frequency of 9.45 GHz, the surface current was concentrated at an electrode, and the electric field was enhanced by the proposed design to lower the response power level of the incident signal. When a low-power signal is injected, plasma is not generated, and the incident wave travels without insertion loss. Double-stage slot structures were utilized to broaden the band-pass characteristics in the frequency domain, and the demonstrated plasma limiter exhibited an insertion loss of 1.01 dB at 9.45 GHz. The xenon gas pressure was optimized with the shortest distance of 100 µm between the upper and lower electrodes to reduce the discharge power of the plasma. In the case of a high-power signal input, as xenon-gas breakdown occurred, the transmitted signal was close to zero, and most of the high-power signal was reflected with a blocking efficiency of 40.55 dB. The demonstrated result will be useful to protect the receiver of a radio detection and ranging system from the high power microwave., (© 2023. The Author(s).)

In a prospective, explorative study, the donor-source difference of haploidentical family (HF), matched sibling (MS), and unrelated donors (UD) was evaluated for the outcome of haematopoietic cell transplantations (HCT) in 101 patients with acute myeloid leukaemia (AML) in complete remission (CR). To eliminate compounding effects, a uniform conditioning regimen containing antithymocyte globulin (ATG) was used. After transplantation, there was a significantly higher cumulative incidence of acute graft-versus-host disease (GVHD) in HF-HCT patients (49%, 7%, and 16% for HF-, MS- and UD-HCT respectively; p < 0.001). A quarter of acute GVHD cases observed in HF-HCT patients occurred within three days of engraftment and were characterized by diffuse skin rash, fever, weight gain, and hypoalbuminaemia. This peri-engraftment acute GVHD was not observed in MS-HCT or UD-HCT patients. Additionally, a significantly higher proportion of HF-HCT patients achieved complete donor chimaerism in the peripheral mononuclear cells at one month (88%, 46%, and 69% for HF-, MS- and UD-HCT respectively; p = 0.001). There was no significant difference in engraftment, chronic GVHD, leukaemia recurrence, non-relapse mortality, and patient survival. In patients with AML in CR who received HCT using ATG-containing conditioning, stronger donor-patient alloreactivity was observed in HF-HCT, in terms of increased acute GVHD and higher likelihood of complete donor chimaerism., (© 2022 British Society for Haematology and John Wiley & Sons Ltd.)

Background and Objectives: The Temperament Evaluation of Memphis, Pisa, Paris and San Diego Autoquestionnaire (TEMPS-A) is designed to assess affective temperaments. The short version of the TEMPS-A (TEMPS-A-SV) has been translated into various languages for use in research and clinical settings. However, no research has been conducted to validate the Korean version of the TEMPS-A-SV in patients with mood disorders. The goal of this study is to evaluate the reliability and validity of the TEMPS-A-SV in Korean mood disorder patients., Materials and Methods: In this cross-sectional retrospective study, a total of 715 patients (267 patients with major depressive disorder, 94 patients with bipolar disorder I, and 354 patients with bipolar disorder II) completed the Korean TEMPS-A-SV. Cronbach's alpha and McDonald's omega were used to assess the reliability. Exploratory factor analysis (EFA) was also performed. Spearman's correlation coefficient was used to examine associations between the five temperaments. The difference in five temperament scores between the gender or diagnosis groups was analyzed, and the correlation between five temperament scores and age was tested., Results: The Korean TEMPS-A-SV displayed good internal consistency (α = 0.65-0.88, ω = 0.66-0.9) and significant correlations between the subscales except one (the correlation between hyperthymic and anxious). Using EFA, a two-factor structure was produced: Factor I (cyclothymic, depressive, irritable, and anxious) and Factor II (hyperthymic). The cyclothymic temperament score differed by gender and the anxious temperament score was significantly correlated with age. All the temperaments, except for irritable temperament, showed significant differences between diagnosis groups., Conclusions: Overall, the results show that the TEMPS-A-SV is a reliable and valid measurement that can be used for estimating Koreans' affective temperaments. However, more research is required on affective temperaments and associated characteristics in people with mood disorders.

Bacterial outer membrane vesicles (OMVs) are small unilamellar proteoliposomes, which are involved in various functions including cell to cell signaling and protein excretion. Here, we have engineered the OMVs of Escherichia coli to nano-scaled bioreactors for the degradation of β-lactam antibiotics. This was exploited by targeting a β-lactamase (i.e., CMY-10) into the OMVs of a hyper-vesiculating E. coli BL21(DE3) mutant. The CMY-10-containing OMVs, prepared from the E. coli mutant cultures, were able to hydrolyze β-lactam ring of nitrocefin and meropenem to a specific rate of 6.6 × 10 -8 and 3.9 × 10 -12 μmol/min/µm 3 of OMV, which is approximately 100 and 600-fold greater than those of E. coli-based whole-cell biocatalsyts. Furthermore, CMY-10, which was encapsulated in the engineered OMVs, was much more stable against temperature and acid stresses, as compared to free enzymes in aqueous phase. The OMV-based nano-scaled reaction system would be useful for the remediation of a variety of antibiotics pollution for food and agricultural industry., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: To evaluate the accuracy of intentional change of the frontal ramal inclination (FRI) from virtual to actual orthognathic surgery., Materials and Methods: A total of 16 patients who underwent orthognathic surgery for correction of facial asymmetry and took cone-beam computed tomography 2 to 3 weeks before surgery (T0) and 3 days after surgery (T1A) were selected. After reorientation of cone-beam computed tomography, the digital imaging and communications in medicine data was converted to StereoLithography format for the merging with dental cast scan using R2GATE software (MegaGen Implant, Daegu, Republic of Korea). During virtual surgery using Geomagic Freeform Plus software (3D Systems, Rock Hill, SC), bilateral FRI discrepancy was corrected (T1V) and 3D-printed surgical wafers and customized metal plates were manufactured using 3D printer (Meg-printer II, MegaGen Implant, Daegu, Republic of Korea) and computer numerical control milling machine (ARDEN, TPS Korea Ltd., Gwangju, Republic of Korea) accordingly. During surgery, FRI correction was done using surgical guide and customized plates. The difference between preoperative and virtually corrected FRI (Δ T1V - T0) and the difference between preoperative and actually surgically corrected FRI (Δ T1A - T0) were measured, and the achieved FRI correction in percentage ([Δ T1A - T0]/[Δ T1V - T0] × 100) was calculated., Results: The mean absolute value of virtual FRI correction (Δ T1V - T0) and actual FRI correction (Δ T1A - T0) were 3.0° and 2.9°, respectively (n = 32). In the increased (medial rotation) FRI group, mean virtual and actual correction amount were 3.2° and 2.8° (n = 17), whereas in the decreased (lateral rotation) group, mean virtual and actual correction amount were -2.7° and -3.1°, respectively (n = 15). No statistically significant difference between virtual and actual measurements were present in either group. The mean achievement rate was 102.8%. Tendency of undercorrection (91.2%) in the increased FRI group and overcorrection (116.0%) in the decreased FRI group were noted., Discussion: With virtual planning and computer-aided design and computer-aided manufacturing-made customized plates, it is possible to correct FRI during orthognathic surgery of patients with facial asymmetry., Competing Interests: The authors report no conflicts of interest., (Copyright © 2021 by Mutaz B. Habal, MD.)

Human Immunodeficiency Virus (HIV) relies on host molecular machinery for replication. Systematic attempts to genetically or biochemically define these host factors have yielded hundreds of candidates, but few have been functionally validated in primary cells. Here, we target 426 genes previously implicated in the HIV lifecycle through protein interaction studies for CRISPR-Cas9-mediated knock-out in primary human CD4+ T cells in order to systematically assess their functional roles in HIV replication. We achieve efficient knockout (>50% of alleles) in 364 of the targeted genes and identify 86 candidate host factors that alter HIV infection. 47 of these factors validate by multiplex gene editing in independent donors, including 23 factors with restrictive activity. Both gene editing efficiencies and HIV-1 phenotypes are highly concordant among independent donors. Importantly, over half of these factors have not been previously described to play a functional role in HIV replication, providing numerous novel avenues for understanding HIV biology. These data further suggest that host-pathogen protein-protein interaction datasets offer an enriched source of candidates for functional host factor discovery and provide an improved understanding of the mechanics of HIV replication in primary T cells., (© 2022. The Author(s).)

This study examined forces leading Korean cities to join the WHO Global Network of Age-Friendly Cities and Communities (AFCC) and implement the WHO guidelines, from the perspectives of multiple streams theory and policy transfer theory. We conducted interviews with stakeholders from six member cities and identified relevant themes using directed content analysis. Political motivation was the strongest factor in seeking Network membership, but the lack of political motivations after gaining membership resulted in lackluster local implementation efforts. The national capital cities may need to take a stronger leadership role to encourage the expansion of the WHO AFCC Network in Asia.

Purpose: To determine whether the RETeval electroretinography (ERG) system can be used to evaluate eyes with dense vitreous hemorrhage (VH)., Methods: This retrospective case series study included 69 eyes of 69 patients with acute dense VH. Flicker ERGs were recorded by the RETeval system, an ERG device with adhesive skin electrodes. We evaluated the flicker ERG amplitudes in eyes with VH and the ratio of the VH eye amplitudes compared with the fellow eye amplitudes for each VH cause., Results: In patients with rhegmatogenous retinal detachment, the amplitude ratios were extremely low (0.08 ± 0.03). To detect rhegmatogenous retinal detachment, the area under the receiver operating characteristic curve was 0.977 (95% confidence interval, 0.943-1.000) (best rhegmatogenous retinal detachment cutoff value, 0.14; sensitivity, 100.0%; and specificity, 95.4%). The flicker ERG amplitude was not significantly correlated with the initial visual acuity (ρ = -0.189, P = 0.120) but was positively correlated with the postoperative visual acuity in eyes with VH (ρ = -0.328, P = 0.006)., Conclusion: The RETeval ERG system was found to be a useful diagnostic option in situations where dense VH precluded fundus examination or posterior vitreous detachment was indistinguishable from rhegmatogenous retinal detachment.

Background: This study presents outcomes of management in graft failure (GF) after allogeneic hematopoietic stem cell transplantation (HCT) and provides prognostic information including rare cases of autologous reconstitution (AR)., Methods: We analyzed risk factors and outcomes of primary and secondary GF, and occurrence of AR in 1,630 HCT recipients transplanted over period of 18 years (January 2000-September 2017) at our center., Results: Primary and secondary GF occurred in 13 (0.80%), and 69 patients (10-year cumulative incidence, 4.5%) respectively. No peri-transplant variables predicted primary GF, whereas reduced intensity conditioning (RIC) regimen (relative risk [RR], 0.97-28.0, P < 0.001) and lower CD34⁺ cell dose (RR, 2.44-2.84, P = 0.002) were associated with higher risk of secondary GF in multivariate analysis. Primary GF demonstrated 100% mortality, in the secondary GF group, the 5-year Kaplan-Meier survival rate was 28.8%, relapse ensued in 18.8%, and AR was observed in 11.6% (n = 8). In survival analysis, diagnosis of aplastic anemia (AA), chronic myeloid leukemia and use of RIC had a positive impact. There were 8 patients who experienced AR, which was rarely reported after transplantation for acute leukemia. Patient shared common characteristics such as young age (median 25 years), use of RIC regimen, absence of profound neutropenia, and had advantageous survival rate of 100% during follow period without relapse., Conclusion: Primary GF exhibited high mortality rate. Secondary GF had 4.5% 10-year cumulative incidence, median onset of 3 months after HCT, and showed 5-year Kaplan-Meier survival of 28.8%. Diagnosis of severe AA and use of RIC was both associated with higher incidence and better survival rate in secondary GF group. AR occurred in 11.6% in secondary GF, exhibited excellent prognosis., Competing Interests: The authors have no potential conflicts of interest to disclose., (© 2021 The Korean Academy of Medical Sciences.)

This study aimed to investigate the effects of mindfulness-based mandala coloring made within nature on individuals with chronic widespread musculoskeletal pain (CWP). Thirty-six participants were randomly allocated. In the experimental group, identical interventions and procedures were administered for each experiment. The control group members were untreated and remained in an urban environment. Overall, the experiment showed significant improvements in tender points ( f = 8.791, p = 0.006), total stress level ( f = 14.570, p = 0.001), depressive symptoms ( f = 15.205, p = 0.001), anger symptoms ( f = 7.263, p = 0.011) and salivary cortisol ( f = 10.619, p = 0.003) in the experimental group. The results reflect that MBMC within nature is effective in reducing pain, psychological stress responses, and cortisol levels in individuals with CWP. The positive results could be a product of the experimental design rather than the treatment itself. A rigorous experimental design provides better understanding of MBMC within nature.

Single-cell RNA sequencing (scRNA-seq) of tissues has revealed remarkable heterogeneity of cell types and states but does not provide information on the spatial organization of cells. To better understand how individual cells function within an anatomical space, we developed XYZeq, a workflow that encodes spatial metadata into scRNA-seq libraries. We used XYZeq to profile mouse tumor models to capture spatially barcoded transcriptomes from tens of thousands of cells. Analyses of these data revealed the spatial distribution of distinct cell types and a cell migration-associated transcriptomic program in tumor-associated mesenchymal stem cells (MSCs). Furthermore, we identify localized expression of tumor suppressor genes by MSCs that vary with proximity to the tumor core. We demonstrate that XYZeq can be used to map the transcriptome and spatial localization of individual cells in situ to reveal how cell composition and cell states can be affected by location within complex pathological tissue., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).)

Purpose: To investigate changes in the foveal avascular zone (FAZ) area and retinal vessel density (VD) in the macula of patients receiving multiple anti-vascular endothelial growth factor (VEGF) therapy for neovascular age-related macular degeneration (N-AMD)., Methods: This study included 54 eyes of 54 treatment-naïve N-AMD patients. Thirty-three eyes were treated with intravitreal aflibercept injections, and 21 eyes were treated with intravitreal ranibizumab injections. Unaffected fellow eyes (54 eyes) were used as controls. All image scans were acquired after the macular architecture had recovered with drying up of the subretinal fluid/hemorrhage after treatment., Results: Both the superficial and deep FAZ areas were significantly larger in the aflibercept group than in the control group. The VD was also significantly reduced in the aflibercept group., Conclusions: Prolonged and repeated anti-VEGF therapy may cause an increase in the FAZ area and a decrease in the VD in patients with N-AMD, indicating ischemic damage.

Aim: We aimed to determine the incidence of and identify the factors associated with treatment-resistant depression (TRD), psychiatric conditions, hospitalization, and cost in patients with major depressive disorder (MDD) who were treated using second-line strategies after an inadequate response to initial antidepressants (AD)., Materials and Methods: Using South Korean National Health Insurance claims data (1 January 2013 to 30 June 2018), we conducted a retrospective cohort analysis in newly treated patients with MDD who subsequently switched or added AD, or added atypical antipsychotics (AAPs) as a second-line treatment. We assessed the incidence of treatment-resistant depression (TRD), psychiatric conditions, and hospitalization for the first 2 years and costs in the third year. Odds ratios (ORs) or relative ratios were estimated using logistic and linear regression models to identify the risk factors for clinical and economic outcomes., Results: In 15,887 patients, the TRD was 16.81% during the 24-month follow-up period (14.14% in switching AD, 19.65% in adding AD, and 19.91% in adding AAP; p  < 0.0001). When adding AD or AAP, the OR of TRD was 1.43 (95% confidence interval (CI): 1.30-1.56) and 1.42 (95% CI: 1.23-1.65), respectively, compared to switching AD. However, these factors were not associated with the incidence of psychiatric conditions. Adding AAP increased hospitalization (OR = 1.25, 95% CI: 1.11-1.41), the number of inpatient days by 2.57-fold (95% CI: 1.75-3.76), and cost by 1.20-fold (95% CI: 1.02-1.40), compared to switching AD; adding AD did not show a significant association with these outcomes., Conclusions: In patients with MDD with inadequate responses to initial AD, TRD still occurred after subsequent treatments according to clinical guidelines. Since the effectiveness of second treatment strategies can differ in reality, further analysis of the clinical and economic evidence regarding second treatment strategies, such as adding AD or AAP, is needed using real-world data.

We investigate retinal layer thickness and capillary vessel density (VD) in the patients with central serous chorioretinopathy (CSC) who recovered spontaneously and evaluate the correlation between the changes in these values and visual outcomes using swept-source optical coherence tomography (SS-OCT) and OCT angiography (OCTA). This retrospective case-control study included 34 eyes of 34 patients with spontaneously resolved acute CSC. The changes in retinal layer thickness and capillary VD were examined using SS-OCT and OCTA after complete resolution of subretinal fluid (SRF). The fellow eyes and 34 healthy eyes were used as controls. In the eyes with CSC, the outer retinal layer was significantly thinner than in the eyes of fellow and healthy controls. The foveal avascular zone area and VDs in the superficial and deep capillary plexus in the eyes with CSC were not significantly different from those in the eyes of fellow and healthy controls. The VD of the choriocapillaris in the eyes with CSC was significantly lower than that in the eyes of fellow and healthy controls. Correlation analyses revealed that the outer retinal layer thickness and initial visual acuity were positively correlated with the final visual acuity. Furthermore, the initial SRF area and height were negatively correlated with the outer retinal layer thickness after SRF resolution. Attenuation of outer retinal layer thickness and decreased VD of the choriocapillaris were observed in the eyes with spontaneously resolved acute CSC. The outer retinal layer thickness could be an important visual predictor of CSC.

Human regulatory T (T reg ) cells are essential for immune homeostasis. The transcription factor FOXP3 maintains T reg cell identity, yet the complete set of key transcription factors that control T reg cell gene expression remains unknown. Here, we used pooled and arrayed Cas9 ribonucleoprotein screens to identify transcription factors that regulate critical proteins in primary human T reg cells under basal and proinflammatory conditions. We then generated 54,424 single-cell transcriptomes from T reg cells subjected to genetic perturbations and cytokine stimulation, which revealed distinct gene networks individually regulated by FOXP3 and PRDM1, in addition to a network coregulated by FOXO1 and IRF4. We also discovered that HIVEP2, to our knowledge not previously implicated in T reg cell function, coregulates another gene network with SATB1 and is important for T reg cell-mediated immunosuppression. By integrating CRISPR screens and single-cell RNA-sequencing profiling, we have uncovered transcriptional regulators and downstream gene networks in human T reg cells that could be targeted for immunotherapies.

Purpose: To compare the properties of the lamina cribrosa (LC) and the peripapillary vessel density between branch retinal vein occlusion (BRVO) and normal-tension glaucoma (NTG), using swept-source optical coherence tomography and optical coherence tomography angiography., Methods: This retrospective study included 21 eyes of 21 patients with BRVO and 43 eyes of 43 patients with NTG who were treated from June 2016 to September 2017. The anterior LC depth (ALCD) and LC thickness (LCT) at the mid-superior, central, and mid-inferior levels; the mean difference in ALCD; and the peripapillary vessel density in the superficial and deep capillary plexuses and the choriocapillaris were compared between groups., Results: ALCD at the mid-superior, central, and mid-inferior levels was significantly greater in the NTG group (P < 0.05), while LCT was comparable between the groups. The mean difference in ALCD was significantly greater in the BRVO group (P = 0.03). The peripapillary vessel density in the superotemporal segment of the superficial capillary plexus was significantly lower in the BRVO group, while the density in all segments of the choriocapillaris was significantly lower in the NTG group (P < 0.05 for all)., Conclusions: Our findings demonstrate that BRVO and NTG have different LC structures and peripapillary vessel densities., Competing Interests: No authors have competing interests.

Appropriate use and interpretation of serological tests for assessments of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) exposure, infection and potential immunity require accurate data on assay performance. We conducted a head-to-head evaluation of ten point-of-care-style lateral flow assays (LFAs) and two laboratory-based enzyme-linked immunosorbent assays to detect anti-SARS-CoV-2 IgM and IgG antibodies in 5-d time intervals from symptom onset and studied the specificity of each assay in pre-coronavirus disease 2019 specimens. The percent of seropositive individuals increased with time, peaking in the latest time interval tested (>20 d after symptom onset). Test specificity ranged from 84.3% to 100.0% and was predominantly affected by variability in IgM results. LFA specificity could be increased by considering weak bands as negative, but this decreased detection of antibodies (sensitivity) in a subset of SARS-CoV-2 real-time PCR-positive cases. Our results underline the importance of seropositivity threshold determination and reader training for reliable LFA deployment. Although there was no standout serological assay, four tests achieved more than 80% positivity at later time points tested and more than 95% specificity.

α-Synuclein (α-Syn) is a key pathogenic protein in α-synucleinopathies including Parkinson disease (PD) and Dementia with Lewy Bodies. The aggregation of α-Syn is believed to be deleterious and a critical step leading to neuronal dysfunction and death. One of the factors that may contribute to the initial steps of this aggregation is crosslinking through transglutaminase 2 (TG2). We previously demonstrated that overexpression of TG2 exacerbates α-Syn toxicity in mice and yeast by increasing the higher-order species of α-Syn. Herein, we investigated whether deletion of the TG2 encoding gene could mitigate the toxicity of α-Syn in vivo. Compared with α-Syn transgenic (Syn Tg ) mice, TG2 null /α-Syn transgenic mice (TG2 KO /Syn Tg ) exhibited a reduced amount of phosphorylated α-Syn aggregates and fewer proteinase K-resistant α-Syn aggregates in sections of brain tissue. Neuritic processes that are depleted in Syn Tg mice compared to wild-type mice were preserved in double TG2 KO /Syn Tg mice. Additionally, the neuroinflammatory reaction to α-Syn was attenuated in TG2 KO /Syn Tg animals. These neuropathological markers of diminished α-Syn toxicity in the absence of TG2 were associated with better motor performance on the rotarod and balance beam. These results suggest that deleting TG2 reduces the toxicity of α-Syn in vivo and improves the behavioral performance of Syn Tg mice. Accordingly, these findings collectively support pharmacological inhibition of TG2 as a potential disease modifying therapeutic strategy for α-synucleinopathies., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The national school uniform remained a requirement for all primary and secondary school students in Malaysia since its first introduction by the British missionaries in the nineteenth century. Although it is commonly thought that wearing uniforms improve cohesion among students, little research has been done to investigate the perception of national school uniforms and perceived cohesion among Malaysian students. The aim of the current study is to explore the experiences of former students, teachers and school counselors in relation to Malaysia's national school uniform policy, their influence on student life, and their relationship to school and national cohesion. Three studies were conducted. In Study 1, Malaysian secondary school graduates ( N = 192) completed a self-report questionnaire with close and open-ended questions. In Study 2 and study 3, secondary school teachers ( N = 10) and school counselors ( N = 6) participated in semi-structured interviews, respectively. Although students felt the uniforms were unattractive, they had neutral to somewhat positive feelings about them overall. They did not report greater school cohesion or national identity related to uniforms but did report that uniforms lessened the importance of socioeconomic and other differences. Teachers and counselors reported similar views; they did not perceive much influence on national or school identity but did see advantages in terms of discipline and the lessening of student inequality. Generally, teachers and counselors were in favor of school uniforms but some suggested that uniform designs could be more culturally inclusive, particularly in regard to Muslim and non-Muslim students. Limitations of using non-representative sampling and future direction for Malaysian school uniforms were discussed. This study highlights the important characteristics that should be taken into consideration by educators and policymakers for future improvement of the national school uniform policy in Malaysia., (Copyright © 2020 Woo, Tam, Bonn and Tagg.)

Regulatory T (T reg ) cells are required to control immune responses and maintain homeostasis, but are a significant barrier to antitumour immunity 1 . Conversely, T reg instability, characterized by loss of the master transcription factor Foxp3 and acquisition of proinflammatory properties 2 , can promote autoimmunity and/or facilitate more effective tumour immunity 3,4 . A comprehensive understanding of the pathways that regulate Foxp3 could lead to more effective T reg therapies for autoimmune disease and cancer. The availability of new functional genetic tools has enabled the possibility of systematic dissection of the gene regulatory programs that modulate Foxp3 expression. Here we developed a CRISPR-based pooled screening platform for phenotypes in primary mouse T reg cells and applied this technology to perform a targeted loss-of-function screen of around 500 nuclear factors to identify gene regulatory programs that promote or disrupt Foxp3 expression. We identified several modulators of Foxp3 expression, including ubiquitin-specific peptidase 22 (Usp22) and ring finger protein 20 (Rnf20). Usp22, a member of the deubiquitination module of the SAGA chromatin-modifying complex, was revealed to be a positive regulator that stabilized Foxp3 expression; whereas the screen suggested that Rnf20, an E3 ubiquitin ligase, can serve as a negative regulator of Foxp3. T reg -specific ablation of Usp22 in mice reduced Foxp3 protein levels and caused defects in their suppressive function that led to spontaneous autoimmunity but protected against tumour growth in multiple cancer models. Foxp3 destabilization in Usp22-deficient T reg cells could be rescued by ablation of Rnf20, revealing a reciprocal ubiquitin switch in T reg cells. These results reveal previously unknown modulators of Foxp3 and demonstrate a screening method that can be broadly applied to discover new targets for T reg immunotherapies for cancer and autoimmune disease.

Systemic lupus erythematosus (SLE) is characterized by impaired clearance of apoptotic cells. Milk fat globule epidermal growth factor 8 (MFGE8) is a protein that connects αvβ3 integrin on phagocytic macrophages with phosphatidylserine on apoptotic cells. We investigated whether genetic variation of the MFGE8 gene and serum MFGE8 concentration are associated with SLE. Single nucleotide polymorphisms (SNPs) were genotyped and serum concentrations were analyzed. The rs2271715 C allele and rs3743388 G allele showed higher frequency in SLE than in healthy subjects (HSs). Three haplotypes were found among 4 SNPs (rs4945, rs1878327, rs2271715, and rs3743388): AACG, CGCG, and CGTC. CGCG haplotype was significantly more common in SLE than in HSs. rs4945 was associated with the erythrocyte sedimentation rate and rs1878327 was associated with alopecia, C-reactive protein, complement 3, anti-dsDNA antibody, and high disease activity. rs2271715 and rs3743388 were associated with renal disease, cumulative glucocorticoid dose, and cyclophosphamide and mycophenolate mofetil use. Serum MFGE8 concentrations were significantly higher in SLE than in HSs. Furthermore, the levels of MFGE8 were significantly higher in SLE than HSs of the rs2271715 CC genotype. In conclusion, MFGE8 genetic polymorphisms are associated not only with susceptibility to SLE but also with disease activity through modulation of gene expression.

Background: The suicide rate of the youth in South Korea has been increasing, and suicide of the youth still has been the most common cause of death since 2007. We aimed to determine the trends and the regional risk factors of youth suicide in South Korea from 2001 to 2010., Subjects and Methods: We used the data from the National Statistical Office to calculate the standardized suicide rates and various regional data including population census, employment, and labor. To calculate the effect of individual risk factors, we used the data from the fourth Korean Youth Risk Behavior Web-based Survey (KYRBWS-VI). Conditional autoregressive model for regional standardized mortality ratio (SMR) using inter-regional spatial information was fitted., Results: Suicide rates of adolescents aged 12 to 18 was from 3.5 per 100,000 people in 2001 and 5.3 per 100,000 in 2010. There were no significant gender difference in suicide rates, however, the number of suicides among adolescents aged 15-18 accounted for four times than those of adolescents ages 12-14. High proportion of late adolescents, higher number of recipients of national basic livelihood, and higher number of adolescents who treated with depression were related to elevated suicide rate of adolescent. Total sleep time of adolescents and regional unemployment rate were negatively associated with the suicide risk of respective regions., Conclusions: Age distribution, economic status, total sleep time, and the number of adolescent patients with depression were different between those in low and in high adolescent suicidal regions in Korea. Our findings suggest that preferential appliance of adolescent suicide prevention program for regions by considering those factors may be important steps to reduce adolescent suicide in Korea.

: (1) Background: Age-related macular degeneration (AMD) is closely related with retinal pigment epithelial (RPE) cell dysfunction. Although the exact pathogenesis of AMD remains largely unknown, oxidative stress-induced RPE damage is believed to be one of the primary causes. We investigated the molecular mechanisms of pentraxin 3 (PTX3) expression and its biological functions during oxidative injury. (2) Methods: Using enzyme-linked immunosorbent assays and real-time reverse transcription-polymerase chain reaction, we analyzed mRNA and protein levels of PTX3 in the presence or absence of oxidative stress inducer, sodium iodate (NaIO 3 ), in primary human H-RPE and ARPE-19 cells. Furthermore, we assessed cell death, antioxidant enzyme expression, and AMD-associated gene expression to determine the biological functions of PTX3 under oxidative stress. (3) Results: NaIO 3 increased PTX3 expression, in a dose- and time-dependent manner, in H-RPE and ARPE-19 cells. We found phosphorylated Akt, a downstream target of the PI3 kinase pathway, phosphor- mitogen-activated protein kinase kinase 1/2 (ERK), and intracellular reactive oxygen species (ROS) were predominantly induced by NaIO 3 . NaIO 3 -induced PTX3 expression was decreased in the presence of phosphoinositide 3 (PI3) kinase inhibitors, ERK inhibitors, and ROS scavengers. Furthermore, NaIO 3 enhanced mRNA expression of antioxidant enzymes such as glucose-6-phosphate dehydrogenase ( G6PDH ), catalase (CAT) , and glutathione S-reductase ( GSR ) in the control shRNA expressing RPE cells, but not in hPTX3 shRNA expressing RPE cells. Interestingly, NaIO 3 did not induce mRNA expression of AMD marker genes, such as complement factor I ( CFI ), complement factor H ( CFH ), apolipoprotein E ( APOE ), and toll-like receptor 4 ( TLR4 ) in hPTX3 shRNA expressing RPE cells. 4) Conclusions: These results suggest that PTX3 accelerates RPE cell death and might be involved in AMD development in the presence of oxidative stress., Competing Interests: The authors declare no conflict of interest.

Background and Objectives: Corneal neovasculariziation (CNV) is a serious vision-threatening complication; however, all therapeutics have their clinical limitations. The aim of this study is to investigate the efficacy of topical rivoceranib compared with topical bevacizumab in a murine model of corneal neovascularization (CNV). Materials and Methods: Murine CNV was induced by means of total de-epithelization and alkali burn. Mice were divided into five groups according to topical treatment: untreated control, phosphate-buffered saline (PBS), 0.1% and 0.5% rivoceranib, and 0.5% bevacizumab. CNV area and index were measured 7 and 14 days after treatment. After corneal tissues were excised at day 14, the blood and lymphatic vessels were quantified by cluster of differentiation 31 (CD31) and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) immunofluorescence, respectively. Results: After 14 days, treatment groups with 0.1% and 0.5% rivoceranib and 0.5% bevacizumab showed a decrease in CNV area and index compared with the untreated and PBS groups (all p < 0.01). Blood and lymphatic vascularization significantly decreased in the 0.5% rivoceranib and 0.5% bevacizumab groups, as measured by CD31 and LYVE1 immunofluorescence. There was no significant difference of vascularization between the 0.5% rivoceranib and bevacizumab groups. Conclusions: Topical application of rivoceranib could effectively decrease CNV equivalent to topical bevacizumab in a murine model.

Long-term dopamine (DA) replacement therapy in Parkinson's disease (PD) leads to the development of abnormal involuntary movements known as l-Dopa-induced dyskinesia (LID). The transcription factor ΔFosB that is highly up-regulated in the striatum following chronic l-Dopa exposure may participate in the mechanisms of altered neuronal responses to DA generating LID. To identify intrinsic effects of elevated ΔFosB on l-Dopa responses, we induced transgenic ΔFosB overexpression in the striatum of parkinsonian nonhuman primates kept naïve of l-Dopa treatment. Elevated ΔFosB levels led to consistent appearance of LID since the initial acute l-Dopa tests. In line with this motor response, striatal projection neurons (SPNs) responded to DA with changes in firing frequency that reversed at the peak of the motor response, and these unstable SPN activity changes in response to DA are typically associated with the emergence of LID. Transgenic ΔFosB overexpression also induced up-regulation of other molecular markers of LID. These results support an autonomous role of striatal ΔFosB in the adaptive mechanisms altering motor responses to chronic DA replacement in PD., Competing Interests: The authors declare no conflict of interest.

Whole-cell biotransformation is one of the promising alternative approaches to microbial fermentation for producing high-value chemicals. Baeyer-Villiger monooxygenase (BVMO)-based Escherichia coli biocatalysts have been engineered to produce industrially relevant C9 chemicals, such as n-nonanoic acid and 9-hydroxynonanoic acid, from a renewable long-chain fatty acid. The key enzyme in the biotransformation pathway (i.e., BVMO from Pseudomonans putida KT2440) was first engineered, using structure modeling-based design, to improve oxidative and thermal stabilities. Using a stable and tunable plasmid (STAPL) system, E. coli host cells were engineered to have increased plasmid stability and homogeneity of the recombinant E. coli population, as well as to optimize the level of BVMO expression. Multi-level engineering of the key enzyme in host cells, allowed recombinant E. coli expressing a fatty acid double-bond hydratase, a long-chain secondary alcohol dehydrogenase, and the engineered BVMO from P. putida KT2440 (i.e., E6BVMO&#95;C302L/M340L), to ultimately produce C9 chemicals (i.e., n-nonanoic acid and 9-hydroxynonanoic acid) from oleic acid, with a yield of up to 6 mmoL/g dry cells. This yield was 2.4-fold greater than the yield in the control strain before engineering. Therefore, this study will contribute to the development of improved processes for the biosynthesis of industrially relevant medium chain fatty acids via whole-cell biocatalysis., (Copyright © 2019 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)

Whole cell biocatalysts can be used to convert fatty acids into various value-added products. However, fatty acid transport across cellular membranes into the cytosol of microbial cells limits substrate availability and impairs membrane integrity, which in turn decreases cell viability and bioconversion activity. Because these problems are associated with the mechanism of fatty acid transport through membranes, a whole-cell biocatalyst that can form caveolae-like structures was generated to promote substrate endocytosis. Caveolin-1 ( CAV1) expression in Escherichia coli increased both the fatty acid transport rate and intracellular fatty acid concentrations via endocytosis of the supplemented substrate. Furthermore, fatty-acid endocytosis alleviated substrate cytotoxicity in E. coli. These traits attributed to bacterial endocytosis resulted in dramatically elevated biotransformation efficiencies in fed-batch and cell-recycle reaction systems when caveolae-forming E. coli was used for the bioconversion of ricinoleic acid (12-hydroxyoctadec-9-enoic acid) to ( Z)-11-(heptanoyloxy) undec-9-enoic acid. We propose that CAV1-mediated endocytosing E. coli represents a versatile tool for the biotransformation of hydrophobic substrates.

Background: Elderly patients with acute myeloid leukemia (AML) have generally had a poor prognosis with unfavorable clinical and biologic disease features. Hypomethylating agents have shown potential for treating medically unfit and elderly patients with AML., Patients and Methods: We compared the outcomes of elderly patients with AML treated with decitabine and intensive chemotherapy (IC)., Results: The data from 107 patients with newly diagnosed AML aged ≥ 65 years were analyzed. The overall response rate was 38.6% and was significantly greater in the IC group than in the decitabine group (65.6% vs. 26.1%; P < .001). With a median follow-up duration of survivors of 14.8 months, the median overall survival (OS) and event-free survival were 12.3 months (95% confidence interval [CI], 10.0-14.7) and 2.0 months (95% CI, 2.0-2.0), respectively, which were not different between the 2 treatment groups. The FLT3-internal tandem duplication mutation (hazard ratio [HR], 2.637; 95% CI, 1.379-5.043; P = .003), complex karyotype (HR, 2.513; 95% CI, 1.258-5.020; P = .009), and peripheral blood blast percentage at diagnosis (HR, 1.983; 95% CI, 1.148-3.422; P = .014) were analyzed as independent prognostic factors for OS. A subgroup analysis for OS showed that IC was superior to decitabine for patients with the FLT3-internal tandem duplication mutation (P = .025) and poor risk cytogenetics, except for -7/del(7q) (P = .005), and decitabine was associated with longer OS for patients with -7/del(7q) (P = .077)., Conclusion: Decitabine showed a similar OS to IC, despite the lower response rate in patients. The clinical outcomes of specific subgroups seemed to differ with different treatment options. Optimal therapeutic approaches for elderly patients with AML should be further examined., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Wireless avionics intra-communication (WAIC) refers to a wireless communication system among electronic components (e.g., sensors and actuators) that are integrated or installed in an aircraft and it is proposed to replace heavy and expensive wired communication cables. Recently, the use of a frequency band (4.2⁻4.4 GHz) for the WAIC (so-called, WAIC band) has been approved by international telecommunication union (ITU). Accordingly, several existing wireless protocols such as IEEE 802.11 and IEEE 802.15 are being considered as candidate techniques for the intra-avionics sensor network. In this paper, we perform a real field experiment to investigate wireless channel characteristics in intra-avionics sensor networks at the WAIC bands by a software-defined radio platform (universal software radio peripheral, USRP) and self-produced monopole antennas for the WAIC band. Through the experiment, we validated the feasibility of IEEE 802.11 protocol for the intra-avionics sensor network at the WAIC band in real aircraft environments. Furthermore, based on the measurement data, we evaluated the bit error rate (BER) performance of multiple antenna techniques where we considered the maximum ratio combining (MRC) for the multi-antenna receiver and the space-time block coding (STBC) for the multi-antenna transmitter.

A persistent concern with CRISPR-Cas9 gene editing has been the potential to generate mutations at off-target genomic sites. While CRISPR-engineering mice to delete a ~360 bp intronic enhancer, here we discovered a founder line that had marked immune dysregulation caused by a 24 kb tandem duplication of the sequence adjacent to the on-target deletion. Our results suggest unintended repair of on-target genomic cuts can cause pathogenic "bystander" mutations that escape detection by routine targeted genotyping assays., Competing Interests: A.M. is a co-founder of Spotlight Therapeutics, Arsenal Biosciences, and Sonoma Biotherapeutics. D.R.S. is a co-founder of Beeline Therapeutics. A.M. has served as an advisor to Juno Therapeutics and is a member of the scientific advisory board for PACT Pharma. The Marson lab has received sponsored research support from Juno Therapeutics, Epinomics and Sanofi and a gift from Gilead. The Marson lab has received funding from the Parker Institute for Cancer Immunotherapy (PICI) and the Innovative Genomics Institute (IGI). Baylor College of Medicine (BCM) and Miraca Holdings have formed a joint venture with shared ownership and governance of the Baylor Genetics (BG), which performs clinical microarray analysis and clinical exome sequencing. J.R.L. serves on the Scientific Advisory Board of the BG. J.R.L. has stock ownership in 23andMe, is a paid consultant for Regeneron Pharmaceuticals, and is a co-inventor on multiple United States and European patents related to molecular diagnostics for inherited neuropathies, eye diseases, and bacterial genomic fingerprinting. The remaining authors declare no competing interests.

Purpose: The purpose of this study was to compare the translucency and color changes of ceramic laminate veneers of different composition following glazing process., Materials and Methods: 10 mm × 10 mm square specimens of 0.6 mm and 1.0 mm thicknesses were fabricated with IPS e.max Press (EM) and IPS e.max ZirPress (ZP) (n=10 per group). The color coordinates (CIE L * a * b * ) of the specimens were recorded with a colorimeter before and after glazing. The color changes and translucency parameter (TP) were calculated. For the comparisons with the composition and thicknesses between the 'not glazed' and 'glazed' groups, statistical analyses were done through paired T-test, independent two-sample T-test, and multiple regression analysis using SPSS 18.0 ( P <.05)., Results: The TP of 0.6 mm EM was higher than that of 0.6 mm ZP. Total color difference (ΔE * ) between bare and glazed specimens of 1.0 mm EM was greater than that of 1.0 mm ZP with statistical significance. Following glazing, specimens from all groups showed statistically significant amount of decrease in L * and a * , and statistically significant increase in b * . The result of multiple regression analysis of EM and ZP showed that ΔL * improved ΔE * ., Conclusion: Within the limitations of present study, we conclude that translucency and color of ceramic laminate veneers change significantly after glazing process, and the nature and amount of changes vary with different compositions.

Objective: To investigate changes in retinal nerve fiber layer, ganglion cell-inner plexiform layer, and choroidal thickness in the macular area in patients with neovascular age-related macular degeneration who received repeated intravitreal ranibizumab and aflibercept treatments., Methods: This retrospective study included 90 eyes of 90 treatment-naive patients. Fifty eyes were treated with intravitreal injections of aflibercept, and 40 were treated with intravitreal injections of ranibizumab. Unaffected fellow eyes (71 eyes) were used as controls. The dosage was one injection per month for 3 consecutive months as an initial treatment. The patients were examined monthly for 6 months following the initial injection. Additional intravitreal injections were given reactively in an optical coherence tomography-guided "pro re nata" protocol. Measurements of the retinal nerve fiber layer, ganglion cell-inner plexiform layer, full retina, and choroidal thickness were simultaneously obtained via swept-source optical coherence tomography in the nine Early Treatment Diabetic Retinopathy Study subfields., Results: The retinal nerve fiber layer thickness in the nine Early Treatment Diabetic Retinopathy Study subfields did not differ significantly among the three study groups (aflibercept vs. ranibizumab vs. control). The ganglion cell-inner plexiform layer thickness was significantly reduced in the aflibercept group, while the choroidal thickness was reduced in both the aflibercept and ranibizumab groups., Conclusion: Excessive long-term vascular endothelial growth factor inhibition by an anti-vascular endothelial growth factor agent that is trapped by neuronal and retinal pigment epithelium cells may adversely affect the function of physiological vascular endothelial growth factor and harm retinal cells and vessels., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Human T cells are central effectors of immunity and cancer immunotherapy. CRISPR-based functional studies in T cells could prioritize novel targets for drug development and improve the design of genetically reprogrammed cell-based therapies. However, large-scale CRISPR screens have been challenging in primary human cells. We developed a new method, single guide RNA (sgRNA) lentiviral infection with Cas9 protein electroporation (SLICE), to identify regulators of stimulation responses in primary human T cells. Genome-wide loss-of-function screens identified essential T cell receptor signaling components and genes that negatively tune proliferation following stimulation. Targeted ablation of individual candidate genes characterized hits and identified perturbations that enhanced cancer cell killing. SLICE coupled with single-cell RNA sequencing (RNA-seq) revealed signature stimulation-response gene programs altered by key genetic perturbations. SLICE genome-wide screening was also adaptable to identify mediators of immunosuppression, revealing genes controlling responses to adenosine signaling. The SLICE platform enables unbiased discovery and characterization of functional gene targets in primary cells., (Copyright © 2018 Elsevier Inc. All rights reserved.)

To investigate the foveal morphological changes and foveal avascular zone (FAZ) area before and after epiretinal membrane (ERM) surgery. Twenty-two eyes with treatment-naive ERM were included in this retrospective study. The central foveal thickness (CFT) and FAZ area were measured via swept-source optical coherence tomography (SS-OCT) and OCT angiography pre- and postoperatively. The unaffected fellow eyes were used as controls. The preoperative superficial FAZ area was significantly smaller in patients (0.08±0.04 mm 2 ) than in controls (0.33±0.09 mm 2 ; P <0.001). The postoperative superficial FAZ (0.12±0.06 mm 2 ) area was significantly greater than the preoperative area ( P <0.001). The preoperative superficial FAZ area was strongly negatively correlated with CFT ( P <0.001, rho =-0.763). ERM induced significant foveal morphological changes and reduction of the superficial FAZ area. Foveal thickness was restored and FAZ area increased postoperatively. However, the process is rather slow and the recovery is incomplete.

Objective: We investigated how mental health awareness among North Korean refugees transformed depending on temporal-spatial context changes., Methods: In 2013, we conducted interviews with 10 refugees (eight women) who had been in South Korea for over a year and performed a qualitative analysis of the change in mental health awareness in the differences between living in North Korea, escape (a related period of forced sojourn in a third country), and settlement in South Korea., Results: We classified 39 concepts into five main categories. The first two categories (while living in North Korea) were "a mindset for the system, but not for individual mental health" and "being confined in a social environment that was indifferent to mental health." A third category appeared during escape: "focusing on survival amid continuity of intense suffering." The final two categories appeared when settling in South Korea: "recognition of mental health amid cultural shock" and "introspection and sorting oneself out.", Conclusion: This qualitative study enabled a better multi-dimensional understanding of the social and cultural aspects involved in improving mental health awareness among North Korean refugees in South Korea. It is desirable to integrate mental health as a part of daily life and to expand training for North Korean settlers.

Purpose: Wide-neck aneurysms (WNAs) associated with a dilated parent artery (PA) are not uncommon morphological abnormalities and usually cause inappropriate wall apposition and incomplete neck coverage of a tubular stent in stent-assisted coiling of aneurysms. We aimed to introduce a fusiform-shaped stent (FSS) and test its effectiveness in treating intracranial WNAs associated with a dilated PA using a three-dimensional (3D) model., Materials and Methods: Two FSS types were designed with the middle one-third segment dilated by 10% (FSS10) and 20% (FSS20) and were compared with the tubular-shaped stent (TSS). A patient-specific 3D WNA model was prototyped and produced, and in vitro stent placement was performed. Angiographic images of the three stent types were analyzed and compared using predetermined parameters., Results: The stent lumens were significantly larger in FSS10 and FSS20 than in TSS in the middle segments (P=0.046), particularly FSS20 (P=0.018). The non-covered area at the ostium tended to be smaller in FSS10 and FSS20 than in TSS, but the difference was not significant (P>0.05). The stent length was significantly longer in FSS10 and FSS20 than in TSS. The stent cell size was significantly larger in FSS than in TSS., Conclusion: Better vessel wall apposition and aneurysmal neck coverage was observed for FSS than for TSS. No significant difference was observed between FSS10 and FSS20.