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Aung, T.

Publications and source records attributed to Aung, T..

At least 19 recordsLinked to original sources

Bruch's Membrane Contributes to the Structural Integrity of the Human Eye

PurposeTo investigate the contribution of the Bruchs membrane and sclera tissues to the overall structural integrity of the ocular wall. MethodsTwenty-three human globes were subjected to biomechanical testing. A piece of sclera measuring 5 x 5 mm was carefully removed at the nasal region, 2 mm away from the optic nerve head. The intraocular pressure was increased at approximately 1 mmHg/s until Bruchs membrane-uvea-retina-tissue layer (BMUR) ruptured. Next, strips of sclera and Bruchs membrane-choriocapillaris (BMC) complex were isolated from the superior fundus region. Uniaxial tension tests were performed at a strain rate of 0.01/s and sampling rate of 15 Hz. The tangent moduli of the BMC and sclera at 0.01, 0.02 and 0.03 strains were compared. ResultsThe rupture pressure of the BMUR was 98.1 {+/-} 21.4 mmHg. The tangent moduli of the BMC at 0.01, 0.02 and 0.03 strains were 2.96 {+/-} 1.44 MPa, 7.68 {+/-} 1.78 MPa and 9.43 {+/-} 2.11 MPa, respectively, and the tangent moduli of the sclera at 0.01, 0.02 and 0.03 strains were 1.09 {+/-} 0.80 MPa, 2.72 {+/-} 1.67 MPa and 5.69 {+/-} 3.27 MPa, respectively. ConclusionThe BMUR was able to sustain relatively high IOP before rupturing. The uniaxial tensile tests showed that the BMC tangent moduli were about 3 times of those of the sclera at strains of 0.01 and 0.02. Although the sclera is approximately 47 times thicker, the BMC is still likely to make a significant contribution (3.51% to 7.42% at strain <0.03) to the overall structural strength of the ocular wall.

bioengineering↗

Hippo signaling differentially regulates distal progenitor subpopulations and their transitional states to construct the mammalian lungs

Lung size control and cell type specification are key unresolved issues. In this study, we have engineered mosaic patterns of Hippo signaling to reveal the developmental potential of SOX9+ progenitors at the distal lung buds. Our results show that the distal SOX9+ subdomain is sufficient to direct lung outgrowth through bifurcation, providing a mechanism for lung size control. Through single-cell analyses, we identify transitional cell states and candidates for promoting cell fates. Moreover, genetic analysis reveals that Hippo signaling induces distinct cell fates at different SOX9+ subdomains to produce the conducting airways and the alveolar epithelium. These results provide the first extensive map of the developmental paths of lung cells. Some of the developmental paths of transitional cell states in mice correspond to those in human lungs. Together, these studies provide mechanistic insight into how Hippo signaling controls the sequential expansion and differentiation of SOX9+ progenitors to construct the mammalian lungs.

developmental biology↗

Pro-endometriosis macrophage release of IL-33 is key for endometriosis pain and lesion formation

Endometriosis is a painful gynecological inflammatory disease affecting up to 10% of females. When released by sensory neurons, calcitonin gene-related peptide (CGRP) shapes immunity, a process known as neuroimmune communication. We previously showed that nociceptor-derived CGRP polarizes macrophages into pro-endometriosis macrophages (PEMs) that mediates endometrial epithelial (endo-epi) cell proliferation and pain. However, the key mediators involved in this PEM-induced cell proliferation were unknown. Using unbiased approaches, we discovered that nociceptor-derived CGRP induces PEMs to produce IL-33. IL-33 binding to its receptor ST2 is key for endometriotic lesion growth and pain during endometriosis in mice as anti-IL-33 antibody treatment reduced evoked and spontaneous pain as well as lesion size. Chemical or genetic ablation of nociceptors or macrophages also resulted in lower levels of lesion IL-33, demonstrating a neuroimmune-driven mechanism for IL-33 production during endometriosis. In humans, we found that IL-33 is correlated with increased number of glands and fibrosis in lesions and that IL-33 expression in macrophages is also associated with genetic risk of endometriosis. We also provided evidence that suggests a dual role for IL-33 in endometriosis, in which, it is initially required for lesion formation and later for lesion maintenance only, and associated pain. Therefore, targeting IL-33/ST2 signaling may effectively treat endometriosis pain.

neuroscience↗

Aging-dependent dysregulation of EXOSC2 is maintained in cancer as a dependency

Reprogramming of aged donor tissue cells into induced pluripotent stem cells (A-iPSC) preserved the epigenetic memory of aged-donor tissue, defined as genomic instability and poor tissue differentiation in our previous study. The unbalanced expression of RNA exosome subunits affects the RNA degradation complex function and is associated with geriatric diseases including premature aging and cancer progression. We hypothesized that the age-dependent progressive subtle dysregulation of EXOSC2 (exosome component 2) causes the aging traits (abnormal cell cycle and poor tissue differentiation). We used embryonic stem cells as a tool to study EXOSC2 function as the aging trait epigenetic memory determined in A-iPSC because these aging traits could not be studied in senesced aged cells or immortalized cancer cells. We found that the regulatory subunit of PP2A phosphatase, PPP2R5E, is a key target of EXOSC2 and this regulation is preserved in stem cells and cancer.

cell biology↗

Dickkopf1 is a Novel Endogenous Ligand for priming NLRP3 Inflammasome in Macrophages via TLR4

Dickkopf1(DKK1) is a quintessential Wnt antagonist and immunomodulator in various inflammatory diseases. The underlying molecular mechanisms of DKK1-mediated immunomodulation remain elusive. Here, we identified TLR4 as a new receptor for DKK1 to activate NF{kappa}B pathway-mediated gene expressions and pyroptosis via NLRP3 inflammasome in human and mouse macrophages. DKK1 employed TLR4 to initiate NF{kappa}B signaling cascade via MyD88. MyD88-TAK1-NF{kappa}B pathway activation by DKK1 increased HIF1, NF{kappa}B, and NLRP3 protein expression levels, leading to pyroptosis. Unlike LPS, DKK1 did not induce IRAK4 phosphorylation, while the interaction between MyD88 and IRAK4 was maintained for downstream signaling activation. DKK1 did not induce IRF3 phosphorylation in the nucleus and failed to induce IFN{beta} gene expression, indicating differential signaling from LPS. DKK1 primed macrophages via TLR4-MyD88, resulting in NLRP3 inflammasome-mediated pyroptosis via Caspase-1 and Gasdermin D maturation with various NLRP3 inflammasome activators, including Nigericin. Our results demonstrated that DKK1 is a novel endogenous priming ligand that augments differential NF{kappa}B pathway from LPS and NLRP3 inflammasome activation via TLR4 in mouse and human macrophages.

immunology↗

High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

Cerebral cavernous malformations (CCMs) are clusters of thin-walled enlarged blood vessels in the central nervous system that are prone to recurrent hemorrhage and can occur in both sporadic and familial forms. The familial form results from loss-of-function variants in the CCM1, CCM2, or CCM3 gene. Despite a better understanding of CCM pathogenesis in recent years, it is still unclear why CCM3 mutations often lead to a more aggressive phenotype than CCM1 or CCM2 variants. By combining high-throughput differentiation of blood vessel organoids from human induced pluripotent stem cells (hiPSCs) with a CCM1, CCM2, or CCM3 knockout, single-cell RNA sequencing, and high-content imaging, we uncovered both shared and distinct functions of the CCM proteins. While there was a significant overlap of differentially expressed genes in fibroblasts across all three knockout conditions, inactivation of CCM1, CCM2, or CCM3 also led to specific gene expression patterns in neuronal, mesenchymal, and endothelial cell populations, respectively. Taking advantage of the different fluorescent labels of the hiPSCs, we could also visualize the abnormal expansion of CCM1 and CCM3 knockout cells when differentiated together with wild-type cells into mosaic blood vessel organoids. In contrast, CCM2 knockout cells showed even reduced proliferation. These observations may help to explain the less severe clinical course in individuals with a pathogenic variant in CCM2 and to decode the molecular and cellular heterogeneity in CCM disease. Finally, the ability to differentiate blood vessel organoids in a 96-well format will further facilitate their use in drug discovery and other biomedical research studies. STATEMENTS AND DECLARATIONSO_ST_ABSConflicts of interest statementC_ST_ABSThe authors declare no competing interests. The here described protocol for high-throughput organoid synthesis has been filed as a patent application at the European Patent Office (Process number: EP24213596.0) Author contribution statementMR, DSk, and UF designed the study. DSk, VS, LM, and RAP performed most of the functional experiments. SH and TA performed the CAM assays. SR performed the immunohistochemical stainings. SB, DSi, DSk, and VS performed the confocal microscopy and high-content imaging analyses. AE, CB, and EMB performed the scRNA sequencing analysis. AW and CAH performed and analyzed the karyotyping of the hiPSC clones. DSk, RAP, VS, KC, MR, and SB analyzed the data. DSk, VS, LM, and MR prepared figures. All authors contributed to the interpretation of the results. DSk, RAP, VS, and MR drafted the manuscript, and all authors contributed to writing. Ethics statementThis study does not involve human participants or animal subjects. Availability of data and materialsAll relevant data are published within the paper and the supplementary files. ScRNA sequencing data can be accessed through the Gene Expression Omnibus (GEO) database (record number: GSE276497).

molecular biology↗

Ablation of LRP6 in alpha-smooth muscle actin-expressing cells abrogates lung inflammation and fibrosis upon bleomycin-induced lung injury

Low-density lipoprotein receptor-related protein 6 (LRP6) is a receptor for Wnt ligands. Tissue fibrosis is a progressive pathological process with excessive extracellular matrix proteins (ECM) deposition. Myofibroblasts, identified by alpha-smooth muscle actin (SMA) expression, play an important role in tissue fibrosis by producing ECM production. Here we found that Wnt antagonist Dickkopf1 (DKK1) induced gene expressions associated with inflammation and fibrosis in lung fibroblasts. We demonstrated that genetic deletion of LRP6 in SMA-expressing cells using Acta2-cre Lrp6fl/fl (Lrp6AKO) mice abrogated bleomycin (BLM)-induced lung inflammation and fibrosis phenotype, suggesting an important role of LRP6 in modulating inflammation and fibrotic processes in the lung. Our results highlight the crucial role of LRP6 in fibroblasts in regulating inflammation and fibrosis upon BLM-induced lung injury.

pathology↗

A meta-analysis of sex differences in neonatal rodent ultrasonic vocalizations and the implication for the preclinical maternal immune activation model

As the earliest measure of social communication in rodents, ultrasonic vocalizations (USVs) in response to maternal separation are critical in preclinical research on neurodevelopmental disorders (NDDs). While sex differences in both USV production and behavioral outcomes are reported, many studies overlook sex as a biological variable in preclinical models of NDDs. We aimed to evaluate sex differences in USV call parameters and to determine if USVs are differently impacted based on sex in the preclinical maternal immune activation (MIA) model. Results indicate that sex differences in USVs vary with developmental stage and are more pronounced in MIA offspring. Specifically, control females exhibited longer call durations than males in early development (up to postnatal day [PND] 8), but this pattern reverses after PND8. MIA leads to a reduction in call numbers for females compared to same-sex controls in early development, with a reversal post-PND8. MIA decreased call duration and increased total call duration in males, but unlike females, developmental stage did not influence these differences. In males, MIA effects varied by species, with decreased call numbers in rats but increased call numbers in mice. The timing of MIA (gestational day [&le;] 12.5 vs. >12.5) did not significantly affect the results. Our findings highlight the importance of considering sex, developmental timing, and species in USVs research. We discuss how analyzing USV call types and incorporating sex as a biological variable can enhance our understanding of neonatal ultrasonic communication and its translational value in NDD research.

neuroscience↗

Iris Morphological and Biomechanical Factors Influencing Angle Closure During Pupil Dilation

PurposeTo use finite element (FE) analysis to assess what morphological and biomechanical factors of the iris and of the anterior chamber are more likely to influence angle narrowing during pupil dilation. MethodsThe study consisted of 1,344 FE models comprising of the cornea, sclera, lens and iris (stroma, sphincter and dilator tissues) to simulate pupil dilation and to assess changes in angle. For each model, we varied the following parameters: anterior chamber depth (ACD = 2 -4 mm) and width (ACW = 10-12 mm), iris convexity (IC = 0-0.3 mm), thickness (IT = 0.3-0.5 mm), stiffness (E = 4-24 kPa) and Poissons ratio (v = 0-0.3), and simulated pupil dilation. We evaluated for the change in anterior chamber angle ({bigtriangleup}{angle}) and the final dilated anterior chamber angles ({angle}f) from baseline to dilation for each parameter. ResultsThe final dilated AC angles decreased with a smaller ACD ({angle}f = 53.4{degrees}{+/-}12.3{degrees} to 21.3{degrees}{+/-}14.9{degrees}), smaller ACW ({angle}f = 48.2{degrees}{+/-}13.5{degrees} to 26.2{degrees}{+/-}18.2{degrees}), larger IT ({angle}f = 52.6{degrees}{+/-}12.3{degrees} to 24.4{degrees}{+/-}15.1{degrees}), larger IC ({angle}f = 45.0{degrees}{+/-}19.2{degrees} to 33.9{degrees}{+/-}16.5{degrees}), larger E ({angle}f = 40.3{degrees}{+/-}17.3{degrees} to 37.4{degrees}{+/-}19.2{degrees}) and larger v ({angle}f = 42.7{degrees}{+/-}17.7{degrees} to 34.2{degrees}{+/-}18.1{degrees}). The change in AC angle increased with larger ACD ({bigtriangleup}{angle} = 9.37{degrees}{+/-}11.1{degrees} to 15.4{degrees}{+/-}9.3{degrees}), smaller ACW ({bigtriangleup}{angle} = 7.4{degrees}{+/-}6.8{degrees} to 16.4{degrees}{+/-}11.5{degrees}), larger IT ({bigtriangleup}{angle} = 5.3{degrees}{+/-}7.1{degrees} to 19.3{degrees}{+/-}10.2{degrees}), smaller IC ({bigtriangleup}{angle} = 5.4{degrees}{+/-}8.2{degrees} to 19.5{degrees}{+/-}10.2{degrees}), larger E ({bigtriangleup}{angle} = 10.9{degrees}{+/-}12.2{degrees} to 13.1{degrees}{+/-}8.8{degrees}) and larger v ({bigtriangleup}{angle} = 8.1{degrees}{+/-}9.4{degrees} to 16.6{degrees}{+/-}10.4{degrees}). ConclusionsThis parametric study offered valuable insights into the factors that could influence angle closure. The morphology of the iris (IT and IC) and its innate biomechanical behavior (E and v) were crucial in influencing the way the iris deformed during dilation, and angle closure was further exacerbated by decreased AC biometry (ACD and ACW).

bioengineering↗

How Myopia and Glaucoma Influence the Biomechanical Susceptibility of the Optic Nerve Head

PurposeWe aimed to assess optic nerve head (ONH) deformations following acute intraocular pressure (IOP) elevations and horizontal eye movements (adduction and abduction) in control eyes, highly myopic (HM) eyes, HM eyes with glaucoma (HMG), and eyes with pathologic myopia alone (PM) or PM with staphyloma (PM+S). MethodsWe studied 282 eyes, comprising of 99 controls, 51 HM, 35 HMG, 21 PM and 75 PM+S eyes. For each eye, we imaged the ONH using spectral-domain optical coherence tomography (OCT) under the following conditions: (1) primary gaze, (2) 20{degrees} adduction, (3) 20{degrees} abduction and (4) primary gaze with acute IOP elevation (to ~35 mmHg) achieved through ophthalmodynamometry. For each OCT volume, we automatically segmented the ONH tissues using deep learning. We performed digital volume correlation (DVC) analysis to compute IOP- and gaze-induced ONH displacements and effective strains (i.e. local deformations). All biomechanical quantities were compared across groups. ResultsUnder IOP elevation, we found that HM eyes exhibited significantly lower strains (3.9 {+/-} 2.4 %) than PM eyes (6.9 {+/-} 5.0%, p < 0.001), HMG eyes (4.7 {+/-} 1.8%, p = 0.04) and PM+S eyes (7.0 {+/-} 5.2%, p < 0.001). Under adduction, we found that HM eyes exhibited significantly lower strains (4.8% {+/-} 2.7%) than PM+S eyes (6.0 {+/-} 3.1%, p = 0.02). We also found significant associations between axial length (or refractive error) and strains - eyes with higher axial length and greater myopia were associated with higher strains. IOP-induced strains were also positively correlated with adduction-induced strains. ConclusionWe found that HMG eyes experienced significantly higher strains under IOP elevations as compared to HM eyes. Additionally, PM+S eyes experienced highest ONH strains as compared to other groups under all biomechanical loads. Our preliminary findings suggest the possibility of using a simple biomechanical test to tease out the susceptibility of HM eyes to further develop glaucoma and/or staphyloma.

bioengineering↗

The Structural Layers of the Porcine Iris Exhibit Inherently Different Biomechanical Properties

PurposeTo isolate the structural components of the ex vivo porcine iris tissue and to determine their biomechanical properties. MethodsThe porcine stroma and dilator tissues were separated, and their dimensions were assessed using optical coherence tomography (OCT). The stroma underwent flow test (n = 32) to evaluate for permeability using Darcys Law ({Delta}P = 2000 Pa, A = 0.0391 mm2), and both tissues underwent stress relaxation experiments ({varepsilon} = 0.5 with initial ramp of {delta}{varepsilon} = 0.1) to evaluate for their viscoelastic behaviours (n = 28). Viscoelasticity was characterised by the parameters {beta} (half width of the Gaussian distribution), {tau}m(mean relaxation time constant), E0 (instantaneous modulus) and E{infty} (equilibrium modulus). ResultsFor the stroma, the hydraulic permeability was 9.49 {+/-} 3.05 x 10-6 mm2/Pa{middle dot}s, and the viscoelastic parameters were {beta} = 2.50 {+/-} 1.40, and {tau}m = 7.43 {+/-} 4.96 s, with the two moduli calculated to be E0= 14.14 {+/-} 6.44 kPa and E{infty} = 6.08 {+/-} 2.74 kPa. For the dilator tissue, the viscoelastic parameters were {beta} = 2.06 {+/-} 1.33 and {tau}m = 1.28 {+/-} 1.27 s, with the two moduli calculated to be E0 = 9.16 {+/-} 3.03 kPa and E{infty} = 5.54 {+/-} 1.98 kPa. ConclusionWe have established a new protocol to evaluate the biomechanical properties of the structural layers of the iris. Overall, the stroma was permeable and exhibited smaller moduli than those of the dilator muscle. An improved characterisation of iris biomechanics may form the basis to further our understanding of angle closure glaucoma.

bioengineering↗

Enhanced Complex Stability and Optimal JAK Geometry are Pivotal for a Potent Type III Interferon Response

Type I and III Interferons (IFN) constitute the host systems first line of defense against viral infections. Although the two families use distinct extracellular receptor complexes, an identical pair of Janus kinases (JAK) activates a similar set of signal transducer and activator of transcription (STATs) through a conserved pathway. Consequently, type I and III IFNs are presumed to activate a largely overlapping set of IFN-stimulated genes (ISGs) and elicit similar biological responses. Therapeutically, type III IFNs are attractive alternatives to type I IFNs due to their innate tissue specificity and lower systemic toxicity. However, a major limitation of type III IFNs is the significantly lower potency of their physiological activities compared to type I IFNs. To evaluate the role of receptor geometry in IFN signaling, we engineered cell lines that express wild-type and mutant IFN{lambda} receptors (IFN{lambda}R1) with rotated intracellular registers with respect to the associated JAK1. Evaluation of downstream signaling and biological activity of type III IFNs in cells with varied JAK-JAK geometries uncovered variant receptors that result in potentiation of type III IFN signaling and all downstream activities. With the combined use of a high-affinity ligand to stabilize the extracellular receptor complex, we found that the optimization of the intracellular JAK-JAK geometry enhances the type III IFN antiviral and anti-proliferative activities by 2- and 3-logs, respectively. In addition to providing a molecular basis for the observed differences in potency between type I and III IFNs, this work provides deeper insights into broader cytokine signaling mechanisms and novel blueprints for modulating cytokine functions to develop next generation antiviral and anticancer therapeutics.

cell biology↗

Integrative deep learning analysis improves colon adenocarcinoma patient stratification at risk for mortality

Colorectal cancers are the fourth most commonly diagnosed cancer and the second leading cancer in number of deaths. Many clinical variables, pathological features, and genomic signatures are associated with patient risk, but reliable patient stratification in the clinic remains a challenging task. Here we assess how image, clinical, and genomic features can be combined to predict risk. We first observe that deep learning models based only on whole slide images (WSIs) from The Cancer Genome Atlas accurately separate high risk (OS<3years, N=38) from low risk (OS>5years, N=25) patients (AUC=0.81{+/-}0.08, 5year survival p-value=2.13e-25, 5year relative risk=5.09{+/-}0.05) though such models are less effective at predicting OS for moderate risk (3years<OS<5years, N=45) patients (5year survival p-value=0.5, 5year relative risk=1.32{+/-}0.09). However, we find that novel integrative models combining whole slide images, clinical variables, and mutation signatures can improve patient stratification for moderate risk patients (5year survival p-value=6.69e-30, 5year relative risk=5.32{+/-}0.07). Our integrative model combining image and clinical variables is also effective on an independent pathology dataset generated by our team (3year survival p-value=1.14e-09, 5year survival p-value=2.15e-05, 3year relative risk=3.25{+/-}0.06, 5year relative-risk=3.07{+/-}0.08). The integrative model substantially outperforms models using only images or only clinical variables, indicating beneficial cross-talk between the data types. Pathologist review of image-based heatmaps suggests that nuclear shape, nuclear size pleomorphism, intense cellularity, and abnormal structures are associated with high risk, while low risk regions tend to have more regular and small cells. The improved stratification of colorectal cancer patients from our computational methods can be beneficial for preemptive development of management and treatment plans for individual patients, as well as for informed enrollment of patients in clinical trials.

pathology↗

Differing Associations between Optic Nerve Head Strains and Visual Field Loss in Normal- and High-Tension Glaucoma Subjects

PurposeTo study the associations between optic nerve head (ONH) strains under intraocular pressure (IOP) elevation with retinal sensitivity in glaucoma subjects. DesignClinic based cross-sectional study. Participants229 subjects with primary open angle glaucoma (subdivided into 115 high tension glaucoma (HTG) subjects and 114 normal tension glaucoma (NTG) subjects). MethodsFor one eye of each subject, we imaged the ONH using spectral-domain optical coherence tomography (OCT) under the following conditions: (1) primary gaze and (2) primary gaze with acute IOP elevation (to approximately 33 mmHg) achieved through ophthalmodynamometry. A 3-dimensional (3D) strain-mapping algorithm was applied to quantify IOP-induced ONH tissue strain (i.e. deformation) in each ONH. Strains in the pre-lamina tissue (PLT)and the retina, the choroid, the sclera and the lamina cribrosa (LC) were associated (using linear regression) with measures of retinal sensitivity from the 24-2 Humphrey visual field test (Carl Zeiss Meditec, Dublin, CA, USA). This was done globally, then locally according to the regionalization scheme of Garway-Heath et al. Main Outcome MeasuresAssociations between ONH strains and values of retinal sensitivity from visual field testing. ResultsFor HTG subjects, we found that (1) there were significant negative linear associations between ONH strains and retinal sensitivity (p<0.001) (on average, a 1% increase in ONH strains corresponded to a decrease in retinal sensitivity of 1.1 dB), (2) high strain regions co-localized with anatomically-mapped regions of high visual field loss, (3) the strongest negative associations were observed in the superior region and in the PLT. In contrast, for NTG subjects, no significant associations between strains and retinal sensitivity were observed except in the supero-temporal region of the LC. ConclusionWe found significant negative associations between IOP-induced ONH strains and retinal sensitivity in a relatively large glaucoma cohort. Specifically, HTG subjects who experienced higher ONH strains were more likely to exhibit lower retinal sensitivities. Interestingly, this trend was in general less pronounced in NTG subjects, which could suggest a distinct pathophysiology between the two glaucoma subtypes.

bioengineering↗

Changes in Iris Stiffness and Permeability in Primary Angle Closure Glaucoma

PurposeTo evaluate the biomechanical properties of the iris by evaluating iris movement during pupil constriction and to compare such properties between healthy and primary angle-closure glaucoma (PACG) subjects. MethodsA total of 140 subjects were recruited for this study. In a dark room, the anterior segments of one eye per subject were scanned using anterior segment optical coherence tomography (AS-OCT, SS-1000 CASIA, Tomey Corporation, Nagoya, Japan) imaging during induced pupil constriction with an external white light source of 1700 lux. Using a custom segmentation code, we automatically isolated the iris segments from the AS-OCT images, which were then discretized and transformed into a three-dimensional point cloud. For each iris, a finite element (FE) mesh was constructed from the point cloud, and an inverse FE simulation was performed to match the clinically observed iris constriction in the AS-OCT images. Through this optimization process, we were able to identify the elastic modulus and permeability of each iris. ResultsFor all 140 subjects (95 healthy and 45 PACG of Indian/Chinese ethnicity, Age: 60.2{+/-}8.7 for PACG subjects and 57.7{+/-}10.1 for healthy subjects), the simulated deformation pattern of the iris during pupil constriction matched well with OCT images. We found that the iris stiffness was higher in PACG than in healthy controls (24.5{+/-}8.4 kPa vs 17.1{+/-}6.6 kPa with 40 kPa of active stress specified in the sphincter region; p < 0.001), whereas iris permeability was lower (0.41{+/-}0.2 mm2/kPa.s vs 0.55{+/-}0.2 mm2/kPa.s; p = 0.142). ConclusionThis study suggests that the biomechanical properties of the iris in PACG are different from those in healthy controls. An improved understanding of the biomechanical behavior of the iris may have implications for the understanding and management of angle-closure glaucoma.

bioengineering↗

Hybrid Derivative of Cathelicidin and Human Beta Defensin-2 Against Gram-Positive Bacteria: A Novel Approach for the Treatment of Bacterial Keratitis

Bacterial keratitis (BK) is a major cause of corneal blindness globally. This study aimed to develop a novel class of antimicrobial therapy, based on human-derived hybrid host defense peptides (HyHDPs), for treating BK. HyHDPs were rationally designed through combination of functional amino acids in parent HDPs, including LL-37 and human beta-defensin (HBD)-1 to -3. Minimal inhibitory concentrations (MICs) and time-kill kinetics assay were performed to determine the concentration- and time-dependent antimicrobial activity and cytotoxicity was evaluated against human corneal epithelial cells and erythrocytes. In vivo safety and efficacy of the most promising peptide was examined in the corneal wound healing and Staphylococcus aureus (ATCC SA29213) keratitis murine models, respectively. A second-generation HyHDP (CaD23), based on rational hybridization of the middle residues of LL-37 and C-terminal of HBD-2, was developed and was shown to demonstrate good efficacy against methicillin-sensitive and methicillin-resistant S. aureus [MIC=12.5-25.0g/ml (5.2-10.4M)] and S. epidermidis [MIC=12.5g/ml (5.2M)], and moderate efficacy against P. aeruginosa [MIC=25-50g/ml (10.4-20.8M)]. CaD23 (at 25g/ml or 2x MIC) killed all the bacteria within 30 mins, which was 8 times faster than amikacin (25g/ml or 20x MIC). After 10 consecutive passages, CaD23 did not develop any antimicrobial resistance (AMR) whereas amikacin, a commonly used treatment for BK, developed significant AMR (i.e. a 32-fold increase in MIC). Pre-clinical murine studies showed that CaD23 (0.5mg/ml) achieved a median reduction of S. aureus bioburden by 94% (or 1.2 log10 CFU/ml) while not impeding corneal epithelial wound healing. In conclusion, rational hybridization of human-derived HDPs has led to generation of a potentially efficacious and safe topical antimicrobial agent for treating Gram-positive BK, with no/minimal risk of developing AMR.

microbiology↗

Effect of Changing Heart Rate on the Ocular Pulse and Dynamic Biomechanical Behavior of the Optic Nerve Head

PurposeTo study the effect of changing heart rate on the ocular pulse and the dynamic biomechanical behaviour of the optic nerve head (ONH) using a comprehensive mathematical model.\n\nMethodsIn a finite element model of a healthy eye, a biphasic choroid consisted of a solid phase with connective tissues and a fluid phase with blood, and the lamina cribrosa (LC) was viscoelastic as characterized by a stress-relaxation test. We applied arterial pressures at 18 ocular entry sites (posterior ciliary arteries) and venous pressures at four exit sites (vortex veins). In the model, the heart rate was varied from 60 bpm to 120 bpm (increment: 20 bpm). We assessed the ocular pulse amplitude (OPA), pulse volume, ONH deformations and the dynamic modulus of the LC at different heart rates.\n\nResultsWith an increasing heart rate, the OPA decreased by 0.04 mmHg for every 10 bpm increase in heart rate. The ocular pulse volume decreased linearly by 0.13 {micro}L for every 10 bpm increase in heart rate. The storage modulus and the loss modulus of the LC increased by 0.014 MPa and 0.04 MPa, respectively, for every 10 bpm increase in heart rate.\n\nconclusionsIn our model, the OPA, pulse volume, and ONH deformations decreased with an increasing heart rate, while the LC became stiffer. The effects of blood pressure / heart rate changes on ONH stiffening may be of interest for glaucoma pathology.\n\nSupportSingapore Ministry of Education, Academic Research Fund, Tier 2 (R-397-000-280-112).\n\nCommercial relationshipNone

bioengineering↗

Optic Nerve Tortuosity and Globe Proptosis in Normal and Glaucoma Subjects

PurposeTo assess the difference in optic nerve tortuosity during eye movements and globe proptosis between primary open angle glaucoma and normal subjects using orbital magnetic resonance imaging.\n\nMethods10 Chinese subjects matched for age, ethnicity and refractive errors were recruited, including five normal controls and five patients with primary open angle glaucoma. All subjects underwent magnetic resonance imaging to assess their optic nerves and globes for three eye positions: primary gaze, adduction and abduction. Optic nerve tortuosity (optic nerve length divided by the distance between two ends) and globe proptosis (maximum distance between cornea and interzygomatic line) were measured from magnetic resonance imaging images.\n\nResultsIn adduction, the tortuosity of normal eyes was significantly larger than that of the glaucomatous eyes. Optic nerve tortuosity in adduction in the control and glaucoma groups were 1.004{+/-}0.003 (mean {+/-} standard deviation) and 1.001{+/-}0.001, respectively (p=0.037). Globe proptosis (primary gaze) in glaucoma subjects (19.14{+/-}2.11 mm) was significantly higher than that in control subjects (15.32{+/-}2.79 mm; p = 0.046).\n\nConclusionsIn this sample, subjects with glaucoma exhibited more taut optic nerves and more protruding eye globes compared to normal eyes. This may impact optic nerve head deformations in anatomically predisposed patients.\n\nPrecisEyes with glaucoma have tauter optic nerves compared with normal eyes, which may exert more force on the optic nerve head tissues during eye movements.

pathology↗