Nanorefractive index transducer using a ring cavity with an internal h-shaped cavity grounded on Fano resonance.

Building on the Fano resonance observation, a new refractive index transducer structure at the nanoscale is proposed in this article, which is a refractive index transducer consisting of a metal-insulator-metal (MIM) waveguide structure coupled with a ring cavity internally connected to an h-shaped structure (RCIhS). Using an analytical method based on COMSOL software and finite element method (FEM), the effect of different geometric parameters of the structure on the trans-mission characteristics of the system is simulated and analyzed, which in turn illustrates the effect of the structural parameters on the output Fano curves. As simulation results show, the internally connected h-shaped structure is an influential component in the Fano resonance. By optimizing the geometrical parameters of the structure, the system finally accomplishes a sensitivity (S) of 2400 nm/RIU and a figure of merit (FOM) of 68.57. The sensor has also been demonstrated in the realm of temperature detection, having tremendous potential for utilization in future nano-sensing and optically integrated systems.

The gut metabolite indole-3-propionic acid activates ERK1 to restore social function and hippocampal inhibitory synaptic transmission in a 16p11.2 microdeletion mouse model.

BACKGROUND: Microdeletion of the human chromosomal region 16p11.2 (16p11.2 + / - ) is a prevalent genetic factor associated with autism spectrum disorder (ASD) and other neurodevelopmental disorders. However its pathogenic mechanism remains unclear, and effective treatments for 16p11.2 + / -  syndrome are lacking. Emerging evidence suggests that the gut microbiota and its metabolites are inextricably linked to host behavior through the gut-brain axis and are therefore implicated in ASD development. Despite this, the functional roles of microbial metabolites in the context of 16p11.2 + / -  are yet to be elucidated. This study aims to investigate the therapeutic potential of indole-3-propionic acid (IPA), a gut microbiota metabolite, in addressing behavioral and neural deficits associated with 16p11.2 + / - , as well as the underlying molecular mechanisms. RESULTS: Mice with the 16p11.2 + / -  showed dysbiosis of the gut microbiota and a significant decrease in IPA levels in feces and blood circulation. Further, these mice exhibited significant social and cognitive memory impairments, along with hyperactivation of hippocampal dentate gyrus neurons and reduced inhibitory synaptic transmission in this region. However, oral administration of IPA effectively mitigated the histological and electrophysiological alterations, thereby ameliorating the social and cognitive deficits of the mice. Remarkably, IPA treatment significantly increased the phosphorylation level of ERK1, a protein encoded by the Mapk3 gene in the 16p11.2 region, without affecting the transcription and translation of the Mapk3 gene. CONCLUSIONS: Our study reveals that 16p11.2 + / -  leads to a decline in gut metabolite IPA levels; however, IPA supplementation notably reverses the behavioral and neural phenotypes of 16p11.2 + / -  mice. These findings provide new insights into the critical role of gut microbial metabolites in ASD pathogenesis and present a promising treatment strategy for social and cognitive memory deficit disorders, such as 16p11.2 microdeletion syndrome. Video Abstract.

Non-lethal exposure to short-wavelength light-emitting diodes modulates tight-junction structure in human corneal epithelial cells via cAMP-dependent signaling.

Light-emitting diodes (LED)-derived lights have been widely used as a medical treatment in photobiomodulation (PBM). However, the PBM effects in ophthalmology are less well investigated. Herein, we explored the effect of LED-generated light on the tight-junction (TJ) formation in human corneal epithelial cells (HCEs). The HCEs were separately exposed to monochromatic LEDs at wavelengths of 365 nm (UVA), 420 nm (violet), 470 nm (blue), 530 nm (green), 590 nm (amber), 660 nm (deep red), and 740 nm (far red) at 10 J/cm2/day for 1 and 2 days. Long-term cultivation of HCEs without LED exposure for up to 14 days was established as a control. The effects of both LED wavelength and culture duration on cell morphology, cAMP-regulated proteins, TJ-associated proteins, and cell growth-associated proteins were also analyzed. Together with the increase in cell number during prolonged cultivation, cAMP, ZO-1, ZO-2, CLDN1, and CLDN4 all increased significantly during long-term cultivation without LED exposure. There was no difference in HCE viability after exposure to all monochromatic LEDs at an accumulated dose of 20 J/cm2. As determined by immunoblotting, UVA, violet, and blue light increased intracellular cAMP, ZO-1, ZO-2, CLDN1, and CLDN4 expression, respectively. UVA and violet, but not blue, light increased PKAreg-pS77 expression. However, none of the other treatments changed the expression of PKAcat-pT197, VASP-pS157, Bax, Bcl-2, or Bcl-xL. Immunofluorescence staining confirmed the formation of TJ structures. The expressions of ZO-1, ZO-2, CLDN1, and CLDN4 as well as TJ structures 2 days following UVA, violet, and blue exposure were similar to those of control cells after 9 days of cultivation. We conclude that short-wavelength LEDs at non-lethal exposure intensities accelerated the formation of TJ structure in HCEs via a cAMP-dependent regulatory cascade.

The extraction and application of antisymmetric characteristics of the cornea during air-puff perturbations.

BACKGROUND: A non-contact tonometer is used to measure intraocular pressure, and studies have primarily relied on apex displacements to assess corneal properties. However, previous studies have overlooked the asymmetric characteristics of lateral corneal perturbations, leading to a gap in understanding of the lateral mechanical properties and its application. METHOD: To investigate these lateral perturbations, we designed an experiment to sequentially record the corneal profiles when two consecutive air-puffs were applied at the center of the same cornea within a short period. Moreover, we used modal decomposition to decompose anterior surface profiles into symmetric and antisymmetric modes to comprehensively analyze the asymmetric characteristics. To extract mechanical properties, we utilized high-pass frequency analysis (>250 Hz) to filter out noise and errors. RESULTS: Symmetric modes between the two consecutive air-puffs exhibited major similarities during vibration; however, antisymmetric modes exhibited minor differences in lateral perturbations of asymmetric vibration. The antisymmetric modes might be related to air-puff misalignment and mechanical properties. Through applying frequency analysis, the mechanical properties could be proven at high frequencies and misalignment shown at low frequencies. Furthermore, we compared the corneal vibration profiles of 259 healthy participants and 50 patients with keratoconus. Their properties showed that the antisymmetric modes of the keratoconus group exhibited a completely opposite direction of deformation compared to that in the healthy group. CONCLUSIONS: Our proposed algorithm not only extracts antisymmetric characteristics but also offers valuable insights into decompose misalignment and mechanical properties of healthy and keratoconus corneas, presenting a new perspective for corneal biomechanics.

High-resolution full-field optical coherence tomography microscope for the evaluation of freshly excised skin specimens during Mohs surgery: A feasibility study.

Histopathology for tumor margin assessment is time-consuming and expensive. High-resolution full-field optical coherence tomography (FF-OCT) images fresh tissues rapidly at cellular resolution and potentially facilitates evaluation. Here, we define FF-OCT features of normal and neoplastic skin lesions in fresh ex vivo tissues and assess its diagnostic accuracy for malignancies. For this, normal and neoplastic tissues were obtained from Mohs surgery, imaged using FF-OCT, and their features were described. Two expert OCT readers conducted a blinded analysis to evaluate their diagnostic accuracies, using histopathology as the ground truth. A convolutional neural network was built to distinguish and outline normal structures and tumors. Of the 113 tissues imaged, 95 (84%) had a tumor (75 basal cell carcinomas [BCCs] and 17 squamous cell carcinomas [SCCs]). The average reader diagnostic accuracy was 88.1%, with a sensitivity of 93.7%, and a specificity of 58.3%. The artificial intelligence (AI) model achieved a diagnostic accuracy of 87.6 ± 5.9%, sensitivity of 93.2 ± 2.1%, and specificity of 81.2 ± 9.2%. A mean intersection-over-union of 60.3 ± 10.1% was achieved when delineating the nodular BCC from normal structures. Limitation of the study was the small sample size for all tumors, especially SCCs. However, based on our preliminary results, we envision FF-OCT to rapidly image fresh tissues, facilitating surgical margin assessment. AI algorithms can aid in automated tumor detection, enabling widespread adoption of this technique.

Effects of cyclosporine on steroid-refractory dry eyes.

PURPOSE: The purpose is to compare the effects of cyclosporine (CsA) on the steroid-refractory mixed-type dry eye (MTDE) and aqueous deficient dry eye (ADDE). MATERIALS AND METHODS: We retrospectively enrolled 71 patients with levels 3 and 4 dry eye that were refractory to artificial tears and topical steroids. The effects were analyzed using the ocular surface disease index (OSDI) and standard patient evaluation of eye dryness (SPEED) questionnaires, Schirmer test II, blink patterns, lipid layer thickness (LLT), meibomian gland expressibility (MGE) and extent of meibomian gland (MG) dropout (meiboscale). RESULTS: MTDE patients (LLT ≤60 nm, n = 38) were younger than those in ADDE (LLT >60 nm, n = 33). Before CsA treatment, they had higher Schirmer scores, less MGE, and a thinner LLT. There was no statistically significant difference in OSDI/SPEED scores between groups. CsA improved the OSDI in the ADDE group but not in the MTDE group. CsA treatment decreased the severity of superficial punctate keratitis (SPK) in both groups, but it significantly decreased partial blinks, total blinks, and partial blink rates in the ADDE group only. CsA did not increase the Schirmer score, LLT, MGE, or meiboscale grade in both groups. CsA could significantly improve subjective symptoms, SPK, and blink patterns in dry eyes refractory to topical steroids, which were more pronounced in ADDE than in MTDE. CONCLUSION: In MTDE cases, concurrent MG treatment modalities, such as intense pulse light and/or thermal pulsation, could be considered to maximize the effects of CsA treatment.

Thermal preconditioning can reduce the incidence of intraoperatively acquired pressure injuries.

Intraoperatively acquired pressure injuries (IAPIs) occur frequently among patients who undergo surgical procedures that last longer than 3 h. Several studies indicated that heat shock proteins (HSPs) play an important role in the protection of stress-induced damages in skin tissues. Hence, the aim of this study was to investigate the potential preventive effect of thermal preconditioning (TPC) on IAPIs in surgical patients and rats and to identify the differentially expressed HSP genes in response to the above treatment. TPC was performed on one group of hairless rats before the model of pressure injuries was established. Subsequently, the size of skin lesions was measured and the expression levels of mRNA and protein of HSPs of the pressured skin were detected by real-time polymerase chain reaction (RT-PCR), western blot, and immunohistochemical staining. For human studies, 118 surgical patients were randomly divided into the TPC group (n = 59) and the control group (n = 59), respectively. The temperature and pressure of sacral skin, as well as the incidence of pressure injury (PI) were detected and compared. In animal studies, TPC significantly reduced both the size and incidence of PI in rats on the second, third and fourth days post treatment. In addition, the expression levels of both mRNA and protein of HSP27 were increased in the TPC group, compared with the control group. Immunohistochemical staining showed that HSP27 was distributed in various types of dermal cells and increased in basal cells. In human studies, a significant reduction (75%) of IAPIs was observed among the patients in the TPC group. TPC can reduce the incidence of PI in rats and humans, and the upregulation of HSP27 may play an important role in this biological progress. Further studies are warranted to explore the molecular mechanism of the preventive effect in PI mediated by HSP27.

Computed Tomography Morphology of Affected versus Unaffected Sides in Patients with Unilateral Primary Acquired Nasolacrimal Duct Obstruction.

BACKGROUND: This study aimed to describe the anatomical details of the bony nasolacrimal duct (BNLD) and adjacent nasal structures by analyzing computed tomography (CT) images, and to investigate their effects on the development of primary acquired nasolacrimal duct obstruction (PANDO). METHODS: A total of 50 patients with unilateral PANDO who underwent dacryocystorhinostomy, with a mean age of 57.96 years, were included. The preoperative CT images were reviewed to measure the anteroposterior and transverse diameters of the BNLD at the entrance and exit levels, as well as the minimum transverse diameter along the tract. The sagittal CT images were analyzed to classify the shape of the bony canals into columnar, funnel, flare, and hourglass. The associated paranasal abnormalities, including nasal septum deviation (NSD), sinusitis, angle between the bony inferior turbinate and medial wall of the maxillary sinus, and mucosal thickness of the inferior turbinate, were investigated. RESULTS: Fifty CT images were analyzed, and all parameters measured on both sides of the BNLD were not significantly different between the PANDO and non-PANDO sides, except for the minimum transverse diameter, which was significantly smaller on the PANDO side (p = 0.002). Columnar-shaped BNLD was the most common on both sides. No significant difference was observed in the incidence of paranasal abnormalities between sides; however, deviation of the septum toward the non-PANDO side was more common (67.9%). CONCLUSIONS: A small minimum transverse diameter of the BNLD may be a risk factor for PANDO. The association between nasal abnormalities and PANDO was not remarkable.

Implication of Adipogenesis-Coupled CRMP2 Functional Profile in Metabolic Homeostasis and Imbalance.

Our previous studies demonstrated that collapsin response mediator protein 2 (CRMP2) is associated with obesity and, in addition, that hyperglycemia-suppressed CRMP2 augments malignant traits of colorectal cancer and is associated with advanced tumor stage. Regulation of CRMP2 profile was further explored in this study using 3T3-L1 pre-adipocyte adipogenesis as a study model for illustrating the roles of CRMP2 in metabolic homeostasis. Hyperglycemia inhibited expression of CRMP2, adipogenic machinery and adipocyte markers. CRMP2 displayed f-CRMP2 (62~66 kDa) and s-CMRP2 (58 kDa) isoforms at the growth arrest phase. Expression of s-CRMP2 was coupled with the mitotic clonal expansion (MCE) phase to direct cell proliferation and rapidly down-regulated in post-mitotic cells. In the late differentiation phase, f-CRMP2 was co-localized with tubulin in the cortical area. Insulin-enhanced CRMP2-glucose transporter 4 (GLUT4) co-localization and CRMP2 puncta on lipid droplets (LDs) suggested participation of CRMP2 in GLUT4 translocation and LD fusion. Collectively, the CRMP2 functional profile must be finely controlled to adjust cytoskeletal stability for meeting dynamic cellular needs. Manipulating the s-CRMP2/f-CRMP2 ratio and thus the cytoskeleton dynamics is anticipated to improve glucose uptake and insulin sensitivity. In summary, our data provide molecular evidence explaining the functions of CRMP2 in physiological, pathological and disease progression in metabolic homeostasis and disorders related to metabolic abnormalities, including cancer.

An Integrated Community-Based Blood Pressure Telemonitoring Program - A Population-Based Observational Study.

Background: Home blood pressure telemonitoring (BPT) has been shown to improve blood pressure control. A community-based BPT program (the Health+ program) was launched in 2015 in an urban area around a medical center. Objectives: To examine the impact of the BPT program on the use of medical resources. Methods: We conducted a retrospective propensity-score (PS)-matched observational cohort study using the National Health Insurance Research Database (NHIRD) 2013-2016 in Taiwan. A total of 9,546 adults with a high risk of cardiovascular disease participated in the integrated BPT program, and 19,082 PS-matched controls were identified from the NHIRD. The primary and secondary outcome measures were changes in 1-year emergency department visit rate, hospitalization rate, duration of hospital stay, and healthcare costs. Results: The number of emergency department visits in the Health+ group significantly reduced (0.8 to 0.6 per year vs. 0.8 to 0.9 per year, p < 0.0001) along with a significant decrease in hospitalization rate (43.7% to 21.3% vs. 42.7% to 35.3%, p < 0.001). The duration of hospital stay was also lower in the Health+ group (4.3 to 3.3 days vs. 5.3 to 6.5 days, p < 0.0001). The annual healthcare costs decreased more in the Health+ group (USD 1642 to 1169 vs. 1466 to 1393 per year, p < 0.001), compared with the controls. Subgroup analysis of the Health+ group revealed that the improvements in outcomes were significantly greater among those who were younger and had fewer comorbidities, especially without diabetes or hypertension. Conclusions: A community-based integrated BPT program may improve patients' health outcomes and reduce healthcare costs.

Low ambient temperature correlates with the severity of dry eye symptoms.

PURPOSE: The symptoms of dry eye disease (DED) are influenced by environmental factors, but the effect of ambient temperature is less certain. Our purpose was to investigate the relationship between the severity of DED symptoms and the ambient temperature. MATERIALS AND METHODS: This retrospective study reviewed the symptom scores, including ocular surface disease index (OSDI) and standardized patient evaluation of eye dryness (SPEED), as well as tear film parameters of first-time DED patients between June 2018 and June 2019. The contribution of tear film parameters and environmental factors, including ambient temperature, humidity, wind speed, and the concentration of air pollutants, to the severity of dry eye symptoms was evaluated by univariate and multivariate linear regression analyses. RESULTS: There were 351 patients included aged 52.8 ± 13.6 years, and 257 (73.2%) were female. The average tear film break-up time, Schirmer test value, and lipid layer thickness were 2.6 ± 0.7 s, 5.5 ± 4.3 mm, and 64.1 ± 6.0 µm, respectively. The average OSDI and SPEED were 41.8 ± 19.8 and 12.1 ± 5.1, respectively. In winter, the patients reported higher OSDI and SPEED. Both scores were significantly correlated with low ambient temperature. Regression analysis showed that low ambient temperature and Schirmer test value contributed to higher OSDI, while low ambient temperature and younger age contributed to higher SPEED. CONCLUSION: Low ambient temperature plays a significant role in DED symptom severity.

Association between Dry Eye Parameters Depends on Tear Components.

How tear components contribute to dry-eye symptoms/signs remains less well-defined. This observational cross-sectional study enrolled 4817 (F/M = 3590/1227) patients. Subjective symptoms were evaluated with the SPEED and OSDI questionnaires. Fluorescein tear breakup time (FTBUT), superficial punctate keratitis (SPK) grading, Schirmer scores, number of expressible meibomian glands (MGE), lipid layer thickness (LLT), blink/partial blink rates and meibography were recorded. Patients were divided into 4 types according to their Schirmer scores and LLT, i.e., Type 1 (N = 1494): Schirmer > 5 mm, LLT > 60 nm; Type 2 (N = 698): Schirmer > 5 mm, LLT ≤ 60 nm; Type 3 (N = 1160): Schirmer ≤ 5 mm, LLT ≤ 60 nm; Type 4 (N = 1465): Schirmer ≤ 5 mm, LLT > 60 nm. Lipid deficiency (LLT ≤ 60 nm) and aqueous deficiency (Schirmer score ≤ 5 mm) were found in 38.6% and 54.5% of patients, respectively. The majority (62.4%) of lipid-deficient patients were also aqueous deficient, while 44.2% of aqueous-deficient patients were also lipid-deficient. Type 3 patients (mixed type) had the highest symptom scores (p = 0.008 and 0.007 for SPEED and OSDI, respectively), more total blinks (p < 0.001) and the shortest FTBUT (p < 0.001). Stepwise multiple regression demonstrated that LLT and Schirmer score were significant contributors to FTBUT in all 4 types. The FTBUT correlated with SPK severity in all 4 types, with Schirmer score in types 1 and 4, and with LLT in type 3 patients. SPK correlated with LLT and MGE in types 1 and 4. Age correlated with dry eye parameters more significantly than sex. Subtyping by aqueous and lipid components facilitates the understanding of dry eye pathophysiology.

Inhibition of Rho-associated protein kinase activity enhances oxidative phosphorylation to support corneal endothelial cell migration.

Corneal endothelial cell (CEC) dysfunction causes corneal edema and severe visual impairment that require transplantation to restore vision. To address the unmet need of organ shortage, descemetorhexis without endothelial keratoplasty has been specifically employed to treat early stage Fuchs endothelial corneal dystrophy, which is pathophysiologically related to oxidative stress and exhibits centrally located corneal guttae. After stripping off central Descemet's membrane, rho-associated protein kinase (ROCK) inhibitor has been found to facilitate CEC migration, an energy-demanding task, thereby achieving wound closure. However, the correlation between ROCK inhibition and the change in bioenergetic status of CECs remained to be elucidated. Through transcriptomic profiling, we found that the inhibition of ROCK activity by the selective inhibitor, ripasudil or Y27632, promoted enrichment of oxidative phosphorylation (OXPHOS) gene set in bovine CECs (BCECs). Functional analysis revealed that ripasudil, a clinically approved anti-glaucoma agent, enhanced mitochondrial respiration, increased spare respiratory capacity, and induced overexpression of electron transport chain components through upregulation of AMP-activated protein kinase (AMPK) pathway. Accelerated BCEC migration and in vitro wound healing by ripasudil were diminished by OXPHOS and AMPK inhibition, but not by glycolysis inhibition. Correspondingly, lamellipodial protrusion and actin assembly that were augmented by ripasudil became reduced with additional OXPHOS or AMPK inhibition. These results indicate that ROCK inhibition induces metabolic reprogramming toward OXPHOS to support migration of CECs.

Dermatochalasis Aggravates Meibomian Gland Dysfunction Related Dry Eyes.

This study aimed to investigate the relationships between subjective symptoms, objective signs, and dermatochalasis severity in dry-eye patients and the effects of lid hygiene on dry-eye parameters. We retrospectively enrolled 2328 patients who underwent dry-eye examinations and classified them into four groups by dermatochalasis severity. The SPEED and OSDI questionnaires were used to evaluate subjective symptoms. LipiView® II interferometry was used to measure lipid-layer thickness (LLT) and blink/incomplete blink rates and perform meibography. A slit-lamp-aided standardized evaluator measured meibomian gland expressibility (MGE). A meiboscale was used to grade meibomian gland dropout. Fluorescein tear-film break-up time (FTBUT) and superficial punctate keratitis (SPK) were recorded. The Schirmer test II with anesthetics was used to evaluate aqueous tear secretion. The effects of lid hygiene were evaluated in 644 patients who underwent second comprehensive examinations. The median age of patients was 55.3 [46.0-66.0] years (76.0% female). Patients with more severe dermatochalasis were less symptomatic and had less MGE, higher meiboscale grades and average LLT. Dermatochalasis severity was significantly associated with MGE and meiboscale grade in the upper lid. There were no significant differences in the Schirmer test, FTBUT, and SPK among the severity groups. Females were older and had higher LLT and less severe dermatochalasis. Lid hygiene significantly decreased subjective symptoms, LLT, and Schirmer results, increased FTBUT, but did not change MGE or meiboscale grades. Dermatochalasis severity participated in the pathophysiology of dry eyes. Lid hygiene significantly improved subjective symptoms and reduced LLT, more significantly in patients with less severe dermatochalasis.

Predictability of 6 Intraocular Lens Power Calculation Formulas in People With Very High Myopia.

Purpose: To investigate the accuracy of 6 intraocular lens (IOL) power calculation formulas in predicting refractive outcomes in extremely long eyes. Setting: Department of Ophthalmology, Far Eastern Memorial Hospital, Taiwan. Design: Retrospective comparative study. Methods: In this retrospective single-center study, we reviewed 70 eyes of 70 patients with axial length (AL) ≥ 28 mm who had received an uneventful 2.2 mm corneal wound phacoemulsification and in-the-bag IOL placement. The actual postoperative refractive results were compared to the predicted refraction calculated with 6 formulas (Haigis, Hoffer Q, Holladay 1, SRK/T, T2, Barrett Universal II formulas) using IOLMaster 500 as optical biometry in the User Group for Laser Interference Biometry (ULIB) constants. Results: Overall, the Haigis and Barrett formulas achieved the lowest level of mean prediction error (PE) and median absolute error (MedAE). Hoffer Q, Holladay 1, SRK/T, and T2 had hyperopic prediction errors (p < 0.05). The Hoffer Q and Holladay 1 had significantly more MedAE between the 6 formulas. After the mean PE was zeroed out, the MedAE had no significant difference between each group. The absolute error tends to be larger in patients with longer AL. The absolute errors were 30.0-37.1% and 60.0-64.3% within 1.0 D of all patients compared to predicted refraction calculated using various formulas. Conclusion: The Haigis and Barrett Universal II formulas had a better success rate in predicting IOL power in high myopic eyes with AL longer than 28 mm using the ULIB constant in this study. The postoperative refractive results were inferior to the benchmark standards, which indicated that the precision of IOL power calculation in patients with high myopia still required improvement.

Repeated cell sorting ensures the homogeneity of ocular cell populations expressing a transgenic protein.

Transgenic proteins can be routinely expressed in various mammalian cell types via different transgenic systems, but the efficiency of transgene expression is constrained by the complex interplay among factors such as the temporal consistency of expression and compatibility with specific cell types, including ocular cells. Here, we report a more efficient way to express an enhanced green fluorescent protein (EGFP) in human corneal fibroblasts, corneal epithelial cells, and conjunctival epithelial cells through a lentiviral expression system. The relative transducing unit criterion for EGFP-expressing pseudovirions was first determined in HEK-293T cells. Homogeneous populations of EGFP-positive and EGFP-negative cells could be isolated by cell sorting. The half-maximal inhibitory concentration (IC50) value for puromycin was calculated according to viability curves for each cell type. The results revealed that cell types differed with respect to EGFP expression efficiency after transduction with the same amount of EGFP-encoding pseudovirions. Using a cell sorter, the homogeneity of EGFP-positive cells reached >95%. In the initial sorting stage, however, the efficiency of EGFP expression in the sorted cells was noticeably reduced after two rounds of sequential culture, but repeated sorting for up to four rounds yielded homogeneous EGFP-positive human corneal fibroblasts that could be maintained in continuous culture in vitro. The sorted EGFP-positive cells retained their proper morphology and cell type-specific protein expression patterns. Puromycin resistance was found to depend on cell type, indicating that the IC50 for puromycin must be determined for each cell type to ensure the isolation of homogeneous EGFP-positive cells. Taken together, repeated cell sorting is an efficient means of obtaining homogeneous populations of ocular cells expressing a transgenic protein during continuous culture without the potential confounding effects of antibiotics.

Characterization of Collapsin Response Mediator Protein 2 in Colorectal Cancer Progression in Subjects with Diabetic Comorbidity.

BACKGROUND: Common demographic risk factors are identified in colorectal cancer (CRC) and type 2 diabetes mellitus (DM), nevertheless, the molecular link and mechanism for CRC-DM comorbidity remain elusive. Dysregulated glycogen synthase kinase-3 beta under metabolic imbalance is suggested to accelerate CRC pathogenesis/progression via regulating collpasin response mediator protein-2 (CRMP2). Accordingly, roles of CRMP2 in CRC and CRC-DM patients were investigated for elucidating the molecular convergence of CRC and DM. METHODS: CRMP2 profile in tumor tissues from CRC and CRC-DM patients was investigated to explore the link between CRC and DM etiology. Meanwhile, molecular mechanism of glucose to regulate CRMP2 profile and CRC characteristics was examined in vitro and in vivo. RESULTS: CRMP2 was significantly lower in tumor lesions and associated with advanced tumor stage in CRC-DM patients. Physiological hyperglycemia suppressed CRMP2 expression/activity and augmented malignant characteristics of CRC cells. Hyperglycemia promotes actin de-polymerization, cytoskeleton flexibility and cell proliferation/metastasis by downregulating CRMP2 profile and thus contributes to CRC disease progression. CONCLUSIONS: This study uncovers molecular evidence to substantiate and elucidate the link between CRC and T2DM, as well as characterizing the roles of CRMP2 in CRC-DM. Accordingly, altered metabolic adaptations are promising targets for anti-diabetic and cancer strategies.

Clinical and Epidemiological Profiles of Retinal Vasculitis in Northern Taiwan.

PURPOSE: To investigate the epidemiological and clinical profiles in retinal vasculitis in an Asian cohort. METHODS: A 5-year retrospective study of 487 uveitis patients in a tertiary referral center at northern Taiwan. RESULTS: Overall, 18.5% of the cases were associated with retinal vasculitis (RV). Sarcoidosis and cytomegalovirus retinitis were two leading diagnoses. Cases with RV were younger (mean: 39.7 years), less unilateral (50%), and had more chorioretinal involvement (95.5%). Definite diagnosis was reached in 86.7% of RV cases, of which 38.5% had infectious uveitis. While sole arteritis was presented in 8.9% of cases, 63.3% showed phlebitis, and 27.8% involved both. Three distinctive patterns were associated with a higher likelihood of infection: nodular sheathing, continuous perivascular sheathing (OR 4.79), and continuous fluorescein leakage (OR 4.11). CONCLUSION: The presence of RV, especially arteritis, is highly suggestive of identifiable etiology. Distinctive clinical patterns help differentiation of infectious versus noninfectious causes.

Munc13-1 is a Ca2+-phospholipid-dependent vesicle priming hub that shapes synaptic short-term plasticity and enables sustained neurotransmission.

During ongoing presynaptic action potential (AP) firing, transmitter release is limited by the availability of release-ready synaptic vesicles (SVs). The rate of SV recruitment (SVR) to release sites is strongly upregulated at high AP frequencies to balance SV consumption. We show that Munc13-1-an essential SV priming protein-regulates SVR via a Ca2+-phospholipid-dependent mechanism. Using knockin mouse lines with point mutations in the Ca2+-phospholipid-binding C2B domain of Munc13-1, we demonstrate that abolishing Ca2+-phospholipid binding increases synaptic depression, slows recovery of synaptic strength after SV pool depletion, and reduces temporal fidelity of synaptic transmission, while increased Ca2+-phospholipid binding has the opposite effects. Thus, Ca2+-phospholipid binding to the Munc13-1-C2B domain accelerates SVR, reduces short-term synaptic depression, and increases the endurance and temporal fidelity of neurotransmission, demonstrating that Munc13-1 is a core vesicle priming hub that adjusts SV re-supply to demand.

Age affects intraocular lens attributes preference in cataract surgery.

PURPOSE: The aim of this study is to analyze the effects of age on intraocular lens (IOL) attributes preference. MATERIALS AND METHODS: We enrolled 4213 eyes that underwent smooth phacoemulsification and IOL implantation between January 2005 and June 2018. Patients were subdivided into six groups according to their ages, i.e.,≤40, 41-50, 51-60, 61-70, 71-80, and ≥ 81 years old. The difference in preference of IOL attributes regarding age, gender, and year of surgery was analyzed separately. The analyzed IOL attributes included asphericity, astigmatism-correction, presbyopia-correction, and blue-blocking function. RESULTS: The patients averaged 68.3 ± 11.6 years old at the time of surgery. There was no significant difference in age between males and females. There were 1980 patients (47.0%) selected aspheric IOL, 822 patients (19.5%) selected multifocal (MF) IOL, 93 patients (2.2%) selected toric IOL, and 859 patients (20.4%) selected blue-blocking IOL. Adoption of aspheric and MF IOL increased significantly during the study (P < 0.001 for both attributes). There were more young patients selected aspheric and MF IOL (P < 0.001 for both), and the change in the trend of adoption over the years was also most significant in the young group (P < 0.001 for both). The proportion of patients that selected blue-blocking IOL decreased significantly after 2011 (P < 0.001). There was no gender preference in aspheric, MF, and toric IOL selection. However, there were more male patients selected blue-blocking IOL (P = 0.018). CONCLUSION: The adoption of IOLs with emerging technologies increased significantly over the years. Younger adults tended to adopt advanced technology IOL more than the older ones.