EC ophthalmology最新文献

Purpose: Optic neuritis occurring in multiple sclerosis (MS) is a disease characterized by chronic inflammation and demyelination in the optic nerve. Although it has been well appreciated that leukocyte infiltration into the optic nerve is an early event during the course of the disease, there has been no study on visualizing and quantifying leukocyte trafficking in the retina during the progression of MS.

Methods: In this study, we generated green fluorescent protein (GFP)⁺ bone marrow chimeric mice, in which GFP-labeled leukocytes facilitate the visualization of their trafficking in the retina. This reporter was then integrated with a well-established rodent model for MS-experimental autoimmune encephalomyelitis (EAE), allowing high resolution in vivo scanning laser ophthalmoscopy (SLO) to track leukocyte movement in the retina in real time. Quantification of leukocyte trafficking was accomplished through Imaris software.

Results: Through SLO, we were able to localize the GFP signal, allowing us to clearly identify leukocytes within the vascular space. We observed more intense leukocyte migration in the retina of EAE mice, exhibiting three distinct movement behaviors: flowing, rolling/crawling and adherent. There was a marked increase in leukocyte rolling and adhesion in retinal vasculature, particularly in the veins and capillaries after induction of EAE. The velocity of rolling leukocytes ranged from 12.0 to 1065.0 μm/sec in the veins as compared to 14.1 to 942.0 in the capillaries. Furthermore, focal areas of recurrent leukocyte adhesion to endothelial surfaces were observed in EAE retinas.

Conclusion: We generated a novel model that makes it possible to non-invasively track leukocyte trafficking in the retina of EAE mice. Our study demonstrates that leukocyte migration in an MS model is distinctly different from the control, suggesting that leukocytes may play a key role in the development of retinal vascular inflammation and optic neuritis during MS, warranting further investigation of the pathological roles of leukocytes in the disease onset and progression.

Purpose: Optic nerve degeneration is a feature of neurodegenerative eye diseases and causes irreversible vision loss. Therefore, understanding the degenerating patterns of the optic nerve is critical to find the potential therapeutic target for optic neuropathy. However, the traditional method of optic nerve degeneration has the limitations of losing spatiotemporal tissue information. Light sheet fluorescence microscopy (LSFM) is a fluorescence microscopy technique that allows capturing 3D images rapidly with a high spatial optical resolution. In this study, we evaluated the availability of LSFM on the optic nerve with NMDA injected Thy1-CFP mice.

Methods: NMDA injected to both eyes of Thy1-CFP mice. After 7 days from the injection, the retina and optic nerve were collected and immunostained with anti-Iba1 antibody. NMDA excitotoxicity induced RGC, and its axon loss and microglial activation in the retina were observed using confocal microscopy. The immunostained optic nerve was completed the optical clearing process with TDE and mounted for LSFM imaging.

Results: We found that retinal flatmounts confirmed significant loss of CFP-expressing RGC and axon degradation and loss in Thy1-CFP mice at 7 days after NMDA injection. Together with these data verifying that NMDA induces RGC and its axon loss, we confirmed that NMDA excitotoxicity induced microglia activation and leukostasis, such as increased microglia number, transform its morphology to ameboid or round, and increase in attached leukocytes in vessels. Using LSFM, we observed that CFP expressing nerve fiber was well organized and arranged parallel in vehicle treated optic nerve, whileas NMDA injected optic nerve showed axon swelling and fragmentation and loss of axon density from the anterior to the posterior regions. Furthermore, LSFM enabled the observation of microglia phenotype transformation in the entire optic nerve. Unlike microglia in vehicle injected optic nerve, microglia in NMDA injected optic nerve displayed larger soma and short process with high Iba1 expression through the entire optic nerve from the anterior to posterior.

Conclusions: In summary, we examined the applicability of the modified optic clearing protocol for the optic nerve and verified it enabled to acquiring of the 3D images of the optic nerve successfully revealing the complex spatial relationships between the axons, microglia and vasculature throughout the entire organ with single acquisitions. With these optimized techniques, we successfully obtained the high-resolution 3D images of NMDA-induced optic neuropathy, including the clues for optic nerve degeneration such as axon swelling, axonal fragmentation, and microglia activation. Overall, we believe that our current study could help understand the pathology of the optic nerve in neurodegenerative diseases, and it will be the basis for translational research.

Purpose: Patients with Parkinson's disease (PD) exhibit unstable tear films. Tear film lipid composition and structure are related to tear film stability and dry eye and tear lipids have not been characterized in people with PD. The aim of this study is to characterize Meibum tear lipids in donors with PD using ¹H-NMR and infrared spectroscopy.

Methods: Three cohorts were compared: meibum from donors with PD (Mp) n = 10, meibum from donors with PD and dry eye (Mpd) n = 3, meibum from donors without PD (Mn) n = 29.

Results: There were no significant differences, P > 0.05, in hydrocarbon branching for Mp compared with Mn. Mn contained twice as much cholesteryl esters compared with Mp, P < 0.0001. The cooperativity of the phase transition was significantly 37% lower for Mp compared with Mn, P < 0.0001. Mpd was much more ordered (stiffer) with compared with Mp and Mn, P < 0.0001.

Conclusion: Changes in meibum lipid composition and structure could be a marker for and/or contribute to increase the susceptibility of dry eye in patients with PD. A less cooperative phase transition for Mp compared with Mn indicates that Mp was more heterogeneous and/or contained more contaminants than Mn. The data support the idea that more ordered lipid contributes to dry eye.

Purpose: Optical coherence tomography angiography (OCT-A) is a newly developed technique to visualize retinal vasculature non-invasively based on interferometry. Although OCT-A has been used clinically, its applications in small animal studies have been limited. This study is designed to develop and demonstrate the feasibility of a protocol for the use of an en-face OCT-based method to visualize and quantify retinal microvasculature in mice that can be used for in vivo assessment of retina ischemia.

Methods: A customized algorithm was developed to extract angiographic profiles of the mouse retina from en-face OCT using an unmodified Bioptigen Envisu R-Class OCT imaging system. En-face OCT images were collected in living animals and then compared to images acquired following termination of blood flow to the retina. The images were processed with ImageJ using the raw file importer. The vessel enhancement algorithm was developed based on a combination of local contrast enhancement, Laplacian of Gaussian peak detection and background subtraction methods. For comparison, fluorescein angiography (FA) was performed using Heidelberg Spectralis^® HRA+OCT imaging system.

Results: By vessel enhancement algorithm, we successfully extracted retinal vasculature and quantified retinal vessel branch points, vascular area and vessel lengths with AngioTool. While the retinal neuronal structure could be simultaneously identified and quantified using B-scan and volumetric OCT run in the annular scanning model, the retinal vasculature in OCT-A was dramatically diminished after the animals were sacrificed, indicating en-face OCT-A signal is a measure of the blood flow.

Conclusions: These studies indicate that a novel approach to extract angiographs from en-face OCT images by utilizing local structure enhancement can be used to provide depth-resolved retinal vasculature distributions. Simultaneous non-invasive analysis of retinal vessels and neurons by OCT-A and OCT may provide a novel approach to characterize retinal ischemia accompanied by neurovascular coupling.

Basic control theory equations are developed showing classical exponential system response of refraction vs. time R(t), with a characteristic system time constant, in response to a negative (-) step change of near work environmental conditions. Statistical analysis of retrospective data from 226 subjects is presented. It is shown that reasonable significance levels p < 0.001 can be obtained from relatively small sample sizes, over short time intervals. Details from preliminary experimental design using reading glasses at the U.S. Naval Academy at Annapolis are discussed. The conclusion is that positive (+) Add lenses, used as reading glasses during study, can prevent the development of myopia of college students in pilot training.