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Corneal haze phenotype in Aldh3a1-null mice: In vivo confocal microscopy and tissue imaging mass spectrometry.
Chen Y, Jester JV, Anderson DM, Marchitti SA, Schey KL, Thompson DC, Vasiliou V
(2017) Chem Biol Interact 276: 9-14
MeSH Terms: Aldehyde Dehydrogenase, Animals, Cornea, Corneal Diseases, Corneal Stroma, Diazepam Binding Inhibitor, Disease Models, Animal, Dynamic Light Scattering, Epithelium, Epithelium, Corneal, Histones, Lens, Crystalline, Lipids, Mice, Mice, Inbred C57BL, Mice, Knockout, Microscopy, Confocal, Phenotype, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Show Abstract · Added May 6, 2017
ALDH3A1 is a corneal crystallin that protects ocular tissues from ultraviolet radiation through catalytic and non-catalytic functions. In addition, ALDH3A1 plays a functional role in corneal epithelial homeostasis by simultaneously modulating proliferation and differentiation. We have previously shown that Aldh3a1 knockout mice in a C57B6/129sV mixed genetic background develop lens cataracts. In the current study, we evaluated the corneal phenotype of Aldh3a1 knockout mice bred into a C57B/6J congenic background (KO). In vivo confocal microscopy examination of KO and wild-type (WT) corneas revealed KO mice to exhibit corneal haze, manifesting marked light scattering from corneal stroma. This corneal phenotype was further characterized by Imaging Mass Spectrometry (IMS) with spatial resolution that revealed a trilayer structure based on differential lipid localization. In these preliminary studies, no differences were observed in lipid profiles from KO relative to WT mice; however, changes in protein profiles of acyl-CoA binding protein (m/z 9966) and histone H4.4 (m/z 11308) were found to be increased in the corneal epithelial layer of KO mice. This is the first study to use IMS to characterize endogenous proteins and lipids in corneal tissue and to molecularly explore the corneal haze phenotype. Taken together, the current study presents the first genetic animal model of cellular-induced corneal haze due to the loss of a corneal crystallin, and strongly supports the notion that ALDH3A1 is critical to cellular transparency. Finally, IMS represents a valuable new approach to reveal molecular changes underlying corneal disease.
Copyright © 2016. Published by Elsevier B.V.
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19 MeSH Terms
Zinc and Manganese Chelation by Neutrophil S100A8/A9 (Calprotectin) Limits Extracellular Aspergillus fumigatus Hyphal Growth and Corneal Infection.
Clark HL, Jhingran A, Sun Y, Vareechon C, de Jesus Carrion S, Skaar EP, Chazin WJ, Calera JA, Hohl TM, Pearlman E
(2016) J Immunol 196: 336-44
MeSH Terms: Adolescent, Adult, Aged, Animals, Aspergillus fumigatus, Biological Transport, Calgranulin A, Calgranulin B, Chelating Agents, Cornea, Disease Models, Animal, Humans, Hyphae, Keratitis, Leukocyte L1 Antigen Complex, Manganese, Mice, Mice, Inbred C57BL, Mice, Knockout, Middle Aged, Neutrophils, Phagocytosis, Pulmonary Aspergillosis, Spores, Fungal, Young Adult, Zinc
Show Abstract · Added February 5, 2016
Calprotectin, a heterodimer of S100A8 and S100A9, is an abundant neutrophil protein that possesses antimicrobial activity primarily because of its ability to chelate zinc and manganese. In the current study, we showed that neutrophils from calprotectin-deficient S100A9(-/-) mice have an impaired ability to inhibit Aspergillus fumigatus hyphal growth in vitro and in infected corneas in a murine model of fungal keratitis; however, the ability to inhibit hyphal growth was restored in S100A9(-/-) mice by injecting recombinant calprotectin. Furthermore, using recombinant calprotectin with mutations in either the Zn and Mn binding sites or the Mn binding site alone, we show that both zinc and manganese binding are necessary for calprotectin's antihyphal activity. In contrast to hyphae, we found no role for neutrophil calprotectin in uptake or killing of intracellular A. fumigatus conidia either in vitro or in a murine model of pulmonary aspergillosis. We also found that an A. fumigatus ∆zafA mutant, which demonstrates deficient zinc transport, exhibits impaired growth in infected corneas and following incubation with neutrophils or calprotectin in vitro as compared with wild-type. Collectively, these studies demonstrate a novel stage-specific susceptibility of A. fumigatus to zinc and manganese chelation by neutrophil-derived calprotectin.
Copyright © 2015 by The American Association of Immunologists, Inc.
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2 Members
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26 MeSH Terms
Efficacy and predictability of soft tissue ablation using a prototype Raman-shifted alexandrite laser.
Kozub JA, Shen JH, Joos KM, Prasad R, Hutson MS
(2015) J Biomed Opt 20: 105004
MeSH Terms: Cornea, Equipment Design, Equipment Failure Analysis, Humans, In Vitro Techniques, Laser Therapy, Lasers, Solid-State, Pilot Projects, Spectrum Analysis, Raman, Treatment Outcome
Show Abstract · Added March 19, 2018
Previous research showed that mid-infrared free-electron lasers could reproducibly ablate soft tissue with little collateral damage. The potential for surgical applications motivated searches for alternative tabletop lasers providing thermally confined pulses in the 6- to-7-µm wavelength range with sufficient pulse energy, stability, and reliability. Here, we evaluate a prototype Raman-shifted alexandrite laser. We measure ablation thresholds, etch rates, and collateral damage in gelatin and cornea as a function of laser wavelength (6.09, 6.27, or 6.43 µm), pulse energy (up to 3 mJ/pulse), and spot diameter (100 to 600 µm). We find modest wavelength dependence for ablation thresholds and collateral damage, with the lowest thresholds and least damage for 6.09 µm. We find a strong spot-size dependence for all metrics. When the beam is tightly focused (~100-µm diameter), ablation requires more energy, is highly variable and less efficient, and can yield large zones of mechanical damage (for pulse energies>1 mJ). When the beam is softly focused (~300-µm diameter), ablation proceeded at surgically relevant etch rates, with reasonable reproducibility (5% to 12% within a single sample), and little collateral damage. With improvements in pulse-energy stability, this prototype laser may have significant potential for soft-tissue surgical applications.
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10 MeSH Terms
Miniature forward-imaging B-scan optical coherence tomography probe to guide real-time laser ablation.
Li Z, Shen JH, Kozub JA, Prasad R, Lu P, Joos KM
(2014) Lasers Surg Med 46: 193-202
MeSH Terms: Animals, Cornea, Lasers, Solid-State, Ophthalmologic Surgical Procedures, Retina, Swine, Tomography, Optical Coherence
Show Abstract · Added March 19, 2018
BACKGROUND AND OBJECTIVE - Investigations have shown that pulsed lasers tuned to 6.1 µm in wavelength are capable of ablating ocular and neural tissue with minimal collateral damage. This study investigated whether a miniature B-scan forward-imaging optical coherence tomography (OCT) probe can be combined with the laser to provide real-time visual feedback during laser incisions.
STUDY DESIGN/METHODS AND MATERIALS - A miniature 25-gauge B-scan forward-imaging OCT probe was developed and combined with a 250 µm hollow-glass waveguide to permit delivery of 6.1 µm laser energy. A gelatin mixture and both porcine corneal and retinal tissues were simultaneously imaged and lased (6.1 µm, 10 Hz, 0.4-0.7 mJ) through air. The ablation studies were observed and recorded in real time. The crater dimensions were measured using OCT imaging software (Bioptigen, Durham, NC). Histological analysis was performed on the ocular tissues.
RESULTS - The combined miniature forward-imaging OCT and mid-infrared laser-delivery probe successfully imaged real-time tissue ablation in gelatin, corneal tissue, and retinal tissue. Application of a constant number of 60 pulses at 0.5 mJ/pulse to the gelatin resulted in a mean crater depth of 123 ± 15 µm. For the corneal tissue, there was a significant correlation between the number of pulses used and depth of the lased hole (Pearson correlation coefficient = 0.82; P = 0.0002). Histological analysis of the cornea and retina tissues showed discrete holes with minimal thermal damage.
CONCLUSIONS - A combined miniature OCT and laser-delivery probe can monitor real-time tissue laser ablation. With additional testing and improvements, this novel instrument has the future possibility of effectively guiding surgeries by simultaneously imaging and ablating tissue.
© 2013 Wiley Periodicals, Inc.
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7 MeSH Terms
Cannabinoid receptor 1 suppresses transient receptor potential vanilloid 1-induced inflammatory responses to corneal injury.
Yang Y, Yang H, Wang Z, Varadaraj K, Kumari SS, Mergler S, Okada Y, Saika S, Kingsley PJ, Marnett LJ, Reinach PS
(2013) Cell Signal 25: 501-11
MeSH Terms: Animals, Arachidonic Acids, Benzoxazines, Calcium, Calcium Channel Blockers, Cell Line, Disease Models, Animal, Endocannabinoids, Epithelial Cells, Epithelium, Corneal, Glycerides, Humans, Immunity, Innate, MAP Kinase Kinase Kinases, Mice, Mice, Knockout, Mitogen-Activated Protein Kinase 8, Morpholines, Naphthalenes, Patch-Clamp Techniques, Pertussis Toxin, Protein Binding, RNA Interference, RNA, Small Interfering, Receptor, Cannabinoid, CB1, Signal Transduction, TRPV Cation Channels, Wound Healing
Show Abstract · Added June 1, 2014
Cannabinoid receptor type 1 (CB1)-induced suppression of transient receptor potential vanilloid type 1 (TRPV1) activation provides a therapeutic option to reduce inflammation and pain in different animal disease models through mechanisms involving dampening of TRPV1 activation and signaling events. As we found in both mouse corneal epithelium and human corneal epithelial cells (HCEC) that there is CB1 and TRPV1 expression colocalization based on overlap of coimmunostaining, we determined in mouse corneal wound healing models and in human corneal epithelial cells (HCEC) if they interact with one another to reduce TRPV1-induced inflammatory and scarring responses. Corneal epithelial debridement elicited in vivo a more rapid wound healing response in wildtype (WT) than in CB1(-/-) mice suggesting functional interaction between CB1 and TRPV1. CB1 activation by injury is tenable based on the identification in mouse corneas of 2-arachidonylglycerol (2-AG) with tandem LC-MS/MS, a selective endocannabinoid CB1 ligand. Suppression of corneal TRPV1 activation by CB1 is indicated since following alkali burning, CB1 activation with WIN55,212-2 (WIN) reduced immune cell stromal infiltration and scarring. Western blot analysis of coimmunoprecipitates identified protein-protein interaction between CB1 and TRPV1. Other immunocomplexes were also identified containing transforming growth factor kinase 1 (TAK1), TRPV1 and CB1. CB1 siRNA gene silencing prevented suppression by WIN of TRPV1-induced TAK1-JNK1 signaling. WIN reduced TRPV1-induced Ca(2+) transients in fura2-loaded HCEC whereas pertussis toxin (PTX) preincubation obviated suppression by WIN of such rises caused by capsaicin (CAP). Whole cell patch clamp analysis of HCEC showed that WIN blocked subsequent CAP-induced increases in nonselective outward currents. Taken together, CB1 activation by injury-induced release of endocannabinoids such as 2-AG downregulates TRPV1 mediated inflammation and corneal opacification. Such suppression occurs through protein-protein interaction between TRPV1 and CB1 leading to declines in TRPV1 phosphorylation status. CB1 activation of the GTP binding protein, G(i/o) contributes to CB1 mediated TRPV1 dephosphorylation leading to TRPV1 desensitization, declines in TRPV1-induced increases in currents and pro-inflammatory signaling events.
Copyright © 2012 Elsevier Inc. All rights reserved.
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1 Members
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28 MeSH Terms
The effect of temperature on the autofluorescence of scattering and non-scattering tissue.
Walsh AJ, Masters DB, Jansen ED, Welch AJ, Mahadevan-Jansen A
(2012) Lasers Surg Med 44: 712-8
MeSH Terms: Animals, Area Under Curve, Cornea, Fiber Optic Technology, Fluorescence, Lasers, Gas, Rats, Scattering, Radiation, Skin, Spectrometry, Fluorescence, Spectrum Analysis, Swine, Temperature
Show Abstract · Added March 7, 2014
BACKGROUND AND OBJECTIVES - With the increasing use of fluorescence in medical applications, a comprehensive understanding of the effect of temperature on tissue autofluorescence is essential. The purpose of this study is to explore the effect of temperature on the fluorescence of porcine cornea and rat skin and determine the relative contributions of irreversible changes in optical properties and in fluorescence yield.
STUDY DESIGN/MATERIALS AND METHODS - Fluorescence, diffuse reflectance, and temperature measurements were acquired from excised porcine cornea and rat skin over a temperature range of 0-80 °C. A dual excitation system was used with a 337 nm pulsed nitrogen laser for the fluorescence and a white light source for the diffuse reflectance measurements. A thermal camera measured tissue temperature. Optical property changes were inferred from diffuse reflectance measurements. The reversibility of the change in fluorescence was examined by acquiring measurements while the tissue sample cooled from the highest induced temperature to room temperature.
RESULTS - The fluorescence intensity decreased with increasing tissue temperature. This fluorescence change was reversible when the tissue was heated to a temperature of 45 °C, but irreversible when heated to a temperature of 80 °C.
CONCLUSION - Auto-fluorescence intensity dependence on temperature appears to be a combination of temperature-induced optical property changes and reduced fluorescence quantum yield due to changes in collagen structure. Temperature-induced changes in measured fluorescence must be taken into consideration in applications where fluorescence is used to diagnose disease or guide therapy.
Copyright © 2012 Wiley Periodicals, Inc.
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13 MeSH Terms
A mouse model of ocular blast injury that induces closed globe anterior and posterior pole damage.
Hines-Beard J, Marchetta J, Gordon S, Chaum E, Geisert EE, Rex TS
(2012) Exp Eye Res 99: 63-70
MeSH Terms: Animals, Anterior Eye Segment, Blast Injuries, Body Weight, Cataract, Corneal Diseases, Disease Models, Animal, Eye Injuries, Female, Intraocular Pressure, Mice, Mice, Inbred BALB C, Posterior Eye Segment, Tomography, Optical Coherence, Visual Acuity, Wounds, Nonpenetrating
Show Abstract · Added January 20, 2015
We developed and characterized a mouse model of primary ocular blast injury. The device consists of: a pressurized air tank attached to a regulated paintball gun with a machined barrel; a chamber that protects the mouse from direct injury and recoil, while exposing the eye; and a secure platform that enables fine, controlled movement of the chamber in relation to the barrel. Expected pressures were calculated and the optimal pressure transducer, based on the predicted pressures, was positioned to measure output pressures at the location where the mouse eye would be placed. Mice were exposed to one of three blast pressures (23.6, 26.4, or 30.4 psi). Gross pathology, intraocular pressure, optical coherence tomography, and visual acuity were assessed 0, 3, 7, 14, and 28 days after exposure. Contralateral eyes and non-blast exposed mice were used as controls. We detected increased damage with increased pressures and a shift in the damage profile over time. Gross pathology included corneal edema, corneal abrasions, and optic nerve avulsion. Retinal damage was detected by optical coherence tomography and a deficit in visual acuity was detected by optokinetics. Our findings are comparable to those identified in Veterans of the recent wars with closed eye injuries as a result of blast exposure. In summary, this is a relatively simple system that creates injuries with features similar to those seen in patients with ocular blast trauma. This is an important new model for testing the short-term and long-term spectrum of closed globe blast injuries and potential therapeutic interventions.
Copyright © 2012 Elsevier Ltd. All rights reserved.
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2 Members
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16 MeSH Terms
BVES regulates EMT in human corneal and colon cancer cells and is silenced via promoter methylation in human colorectal carcinoma.
Williams CS, Zhang B, Smith JJ, Jayagopal A, Barrett CW, Pino C, Russ P, Presley SH, Peng D, Rosenblatt DO, Haselton FR, Yang JL, Washington MK, Chen X, Eschrich S, Yeatman TJ, El-Rifai W, Beauchamp RD, Chang MS
(2011) J Clin Invest 121: 4056-69
MeSH Terms: Adenocarcinoma, Adherens Junctions, Animals, Cell Adhesion Molecules, Cell Line, Tumor, Colonic Neoplasms, Colonic Polyps, Colorectal Neoplasms, DNA Methylation, Epithelial-Mesenchymal Transition, Epithelium, Corneal, Gene Expression, Gene Silencing, Humans, Membrane Proteins, Mice, Mice, Nude, Muscle Proteins, Mutation, Neoplasm Transplantation, Promoter Regions, Genetic, Signal Transduction, Tight Junctions, Transplantation, Heterologous
Show Abstract · Added June 14, 2013
The acquisition of a mesenchymal phenotype is a critical step in the metastatic progression of epithelial carcinomas. Adherens junctions (AJs) are required for suppressing this epithelial-mesenchymal transition (EMT) but less is known about the role of tight junctions (TJs) in this process. Here, we investigated the functions of blood vessel epicardial substance (BVES, also known as POPDC1 and POP1), an integral membrane protein that regulates TJ formation. BVES was found to be underexpressed in all stages of human colorectal carcinoma (CRC) and in adenomatous polyps, indicating its suppression occurs early in transformation. Similarly, the majority of CRC cell lines tested exhibited decreased BVES expression and promoter DNA hypermethylation, a modification associated with transcriptional silencing. Treatment with a DNA-demethylating agent restored BVES expression in CRC cell lines, indicating that methylation represses BVES expression. Reexpression of BVES in CRC cell lines promoted an epithelial phenotype, featuring decreased proliferation, migration, invasion, and anchorage-independent growth; impaired growth of an orthotopic xenograft; and blocked metastasis. Conversely, interfering with BVES function by expressing a dominant-negative mutant in human corneal epithelial cells induced mesenchymal features. These biological outcomes were associated with changes in AJ and TJ composition and related signaling. Therefore, BVES prevents EMT, and its epigenetic silencing may be an important step in promoting EMT programs during colon carcinogenesis.
2 Communities
5 Members
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24 MeSH Terms
Bves modulates tight junction associated signaling.
Russ PK, Pino CJ, Williams CS, Bader DM, Haselton FR, Chang MS
(2011) PLoS One 6: e14563
MeSH Terms: CCAAT-Enhancer-Binding Proteins, Cell Adhesion Molecules, Cell Line, Epithelial Cells, Epithelium, Corneal, Heat-Shock Proteins, Humans, Membrane Proteins, Muscle Proteins, Signal Transduction, Tight Junctions, rhoA GTP-Binding Protein
Show Abstract · Added March 5, 2014
Blood vessel epicardial substance (Bves) is a transmembrane adhesion protein that regulates tight junction (TJ) formation in a variety of epithelia. The role of TJs within epithelium extends beyond the mechanical properties. They have been shown to play a direct role in regulation of RhoA and ZONAB/DbpA, a y-box transcription factor. We hypothesize that Bves can modulate RhoA activation and ZONAB/DbpA activity through its regulatory effect on TJ formation. Immortalized human corneal epithelial (HCE) cells were stably transfected with Flag-tagged full length chicken Bves (w-Bves) or C-terminus truncated Bves (t-Bves). We found that stably transfected w-Bves and t-Bves were interacting with endogenous human Bves. However, interaction with t-Bves appeared to disrupt cell membrane localization of endogenous Bves and interaction with ZO-1. w-Bves cells exhibited increased TJ function reflected by increased trans-epithelial electrical resistance, while t-Bves cells lost TJ protein immunolocalization at cell-cell contacts and exhibited decreased trans-epithelial electrical resistance. In parental HCE and w-Bves cells ZONAB/DbpA and GEF-H1 were seen at cell borders in the same pattern as ZO-1. However, expression of t-Bves led to decreased membrane localization of both ZONAB/DbpA and GEF-H1. t-Bves cells had increased RhoA activity, as indicated by a significant 30% increase in FRET activity compared to parental HCE cells. ZONAB/DbpA transcriptional activity, assessed using a luciferase reporter probe, was increased in t-Bves cells. These studies demonstrate that Bves expression and localization can regulate RhoA and ZONAB/DbpA activity.
1 Communities
3 Members
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12 MeSH Terms
Long-term results of combined endoscope-assisted pars plana vitrectomy and glaucoma tube shunt surgery.
Tarantola RM, Agarwal A, Lu P, Joos KM
(2011) Retina 31: 275-83
MeSH Terms: Adult, Aged, Aged, 80 and over, Chronic Disease, Corneal Opacity, Endoscopy, Female, Fibrosis, Follow-Up Studies, Glaucoma Drainage Implants, Glaucoma, Angle-Closure, Humans, Intraocular Pressure, Iris, Male, Middle Aged, Prosthesis Implantation, Retrospective Studies, Treatment Outcome, Visual Acuity, Vitrectomy
Show Abstract · Added March 19, 2018
PURPOSE - To assess outcomes after endoscope-assisted pars plana vitrectomy with concurrent pars plana tube shunt placement.
METHODS - Records of 18 adult patients (19 eyes) with uncontrolled chronic angle-closure glaucoma associated with corneal opacification or fibrosed pupils were retrospectively reviewed. All eyes underwent endoscope-assisted pars plana vitrectomy with Baerveldt tube shunt placement into the vitreous cavity between 1997 and 2005. Intraocular pressure reduction, glaucoma medication reduction, complications, and visual acuity were analyzed.
RESULTS - Mean follow-up duration was 62 months (range, 10-106 months). Mean preoperative intraocular pressure was 31.3 ± 10.5 mmHg on 3.4 ± 1.0 glaucoma medications. Intraocular pressure was significantly reduced at each postoperative time point examined. In the 17 eyes without phthisis, intraocular pressure was significantly reduced at the final follow-up examination to a mean of 11.4 ± 2.9 mmHg (P < 0.0001) on 1.3 ± 1.2 medications (P < 0.0001). No complications occurred in 14 of 19 eyes. Postoperatively, best-attained visual acuity improved in 14 of 19 eyes, remained unchanged in 4 of 19 eyes, and was reduced in 1 of 19 eyes.
CONCLUSION - Combined endoscope-assisted pars plana vitrectomy with placement of a Baerveldt tube shunt into the vitreous cavity is a useful intervention in patients with uncontrolled chronic angle-closure glaucoma, media opacities, and limited surgical options.
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21 MeSH Terms