Beta-adrenergic glaucoma drugs reduce lymphatic clearance from the eye: A sequential photoacoustic imaging study.

Our study aims to determine whether the beta-adrenergic system is involved in the regulation of lymphatic drainage from the eye. For this purpose, we assessed the effect of 2 topical beta-adrenergic blockers, timolol and betaxolol, commonly used as glaucoma drugs, on lymphatic clearance of albumin from the aqueous humor to neck lymph nodes. Adult mice were treated with either topical timolol, a non-selective ß-blocker, 0.5% (n = 8), or topical betaxolol, a selective ß1-adrenergic blocker, 0.5% (n = 6) twice daily for 14 days and compared to respective control groups (n = 5 and n = 7). Changes in lymphatic clearance from the eye were assessed using a quantitative in vivo photoacoustic imaging approach. In all subjects, right eye and neck lymph nodes were longitudinally assessed by sequential photoacoustic imaging just prior to near-infrared dye injection into the anterior chamber of the eye, and 20 min, 2 and 4 h after injection. Repeat measurements of mean pixel intensities (MPIs) of right eyes and nodes were performed at all timepoints. The areas under the curves (AUC) were calculated and the AUC of the treated-group was compared to that of controls using the Mann-Whitney U test. The slopes of MPI of each region of interest over time were compared using the linear mixed model after adjusting for IOP decrease after treatment and other parameters such as sex and body weight. In the timolol-treated group, right neck nodes showed significant decrease in AUC signal intensity compared with controls (P = 0.003), and significant decrease in slope of MPI compared with controls (P = 0.0025). In the betaxolol-treated group, right neck nodes showed significant decrease in AUC signal intensity compared with controls (P = 0.02), and significant decrease in slope of MPI compared with controls (P = 0.0069). Topical treatment with timolol and betaxolol reduced lymphatic clearance of albumin from the aqueous humor to the neck lymph nodes. This finding may be relevant for the management of secondary glaucomas and inflammatory eye disease in which the clearance of accumulated proteins and antigen from the eye is important to disease recovery and sight protection. This study suggests that the beta-adrenergic system plays a role in the regulation of lymphatic clearance from the eye.

Age-related decline of lymphatic drainage from the eye: A noninvasive in vivo photoacoustic tomography study.

We aim to determine whether lymphatic drainage from the eye changes with age. Using quantitative photoacoustic tomography, groups of young and older mice were studied in the live state. 10 CD-1 mice of 2-3 months (5M/5F) were studied in addition to 13 older mice of 12-13 months (6M/7F). In each of 23 mice, near-infrared tracer (a near-infrared dye, QC-1 conjugated with Bovine Serum Albumin) was injected into the right eye, and imaging of ipsilateral cervical lymph nodes was performed with laser pulses at 11 different wavelengths prior to and 20 min, 2, 4 and 6 h after injection. Mean pixel intensities (MPIs) of nodes were calculated at each imaging session. The areas under the curves (AUC) were calculated for both groups of mice and compared using the t-test. The slopes of MPI of each region of interest were compared using the linear mixed model before and after adjusting for sex, body weight and intraocular pressure of the right eye. The mean intraocular pressure of right eyes before injection was similar in older and younger groups (12.77 ± 2.01 mmHg and 12.90 ± 2.38 mmHg, respectively; p = 0.888). In each mouse, the photoacoustic signal was detected in the right cervical lymph nodes at the 2-h time point following tracer injection into the right eye. At the 4 and 6 h imaging times, a steady increase of tracer signal was observed. Areas under the curve in the right cervical nodes were decreased significantly in older mice compared to younger mice (p = 0.007). The slopes of MPI in the nodes were significantly decreased in old mice compared to young mice both before and after adjusting for sex, body weight and intraocular pressure of the right eye (p = 0.003). In conclusion, lymphatic drainage from the eye is significantly reduced in older eyes. This finding suggests that impaired lymphatic clearance of aqueous humor, proteins and antigens from the eye may contribute to age-related disease of the eye such as glaucoma and inflammatory eye disease.

Non-invasive dynamic assessment of conjunctival melanomas by photoacoustic imaging.

This study describes non-invasive photoacoustic imaging to detect and monitor the growth of conjunctival melanomas in vivo. Conjunctival melanomas were induced by injection of melanotic B16F10 cells into the subconjunctival space in syngeneic albino C57BL/6 mice. Non-invasive in vivo photoacoustic tomography was performed before, and after tumor induction up to 2 weeks. Spectral unmixing was performed to determine the location and to assess the distribution of melanin. The melanin photoacoustic signal intensity was quantified from the tumor-bearing and control eyes at all timepoints. For postmortem validation, total tumor and melanotic tumor volumes were measured using H&E stained tumor sections and were compared to in vivo photoacoustic imaging measurements. Photoacoustic imaging non-invasively detected eyes bearing conjunctival tumors of varying sizes. The melanin signal was detected as early as immediately following injection of melanotic tumor cells. Changes in tumor size over time were assessed with changes in the volume and intensity of the melanin signal. Four growing tumors and one regressing tumor were observed. Three tumors without significant change in signal intensity over time were observed, showing variable growth. Photoacoustic melanin signal on the last day of in vivo imaging correlated with postmortem total tumor volume (R2 = 0.81) and melanotic tumor volume (R2 = 0.80). The results of our study show that actively growing conjunctival melanomas can be quantified in a non-invasive manner using in vivo photoacoustic tomography. The photoacoustic melanin signal intensity correlated with total and melanotic tumor volume. This novel in vivo imaging platform may help to assess new treatment modalities to manage ocular tumors.

A framework to explore the visual brain in glaucoma with lessons from models and man.

Vision loss in glaucoma is associated with death of retinal ganglion cells. High intraocular pressure is a major risk factor for vision loss from glaucoma, and lowering eye pressure is the goal of all available medical and surgical treatments. Taking a bold step forward, the restoration of vision after severe glaucoma damage is a new Audacious Goal established by National Eye Institute (Sieving, 2012). This means that retinal ganglion cell repair, and replacement, must be considered in the context of visual function restoration. To restore visual function, retinal ganglion cells, after long-distance axonal growth and guidance, should connect to specific target neurons in subcortical visual structures. At the time of the establishment of these connections, the fate of target cells is critical along with the health of retinal ganglion cells. In fact, several lines of evidence demonstrate glaucomatous neural degeneration occurs throughout the central visual system where most information processing takes place. Evidence from multiple studies in experimental glaucoma models, human autopsy cases and neuroimaging studies point to the degeneration of neurons in the lateral geniculate nucleus, a subcortical hub of functional connectivity between the eye and the visual cortex. Maintaining and re-establishing connections of retinal ganglion cells to target neurons in major visual structures is a key endpoint for regenerative medicine strategies. This paper critically reviews studies of visual brain changes in man and experimental animal models, and discusses key factors in the experimental design that are relevant to restoring vision loss in human disease.

Saccadic eye movements in mild traumatic brain injury: a pilot study.

OBJECTIVE: To investigate whether repeat saccadic reaction time (SRT) measurements using a portable saccadometer is useful to monitor patients with mild traumatic brain injury (mTBI). METHODS: Seven patients with newly-diagnosed mTBI and five agematched controls were prospectively recruited from an emergency Department. Saccadic eye movements, symptom self-reporting and neuropsychological tests were performed within one week of injury and again at follow-up three weeks post-injury. Control patients underwent saccade recordings at similar intervals. RESULTS: Median saccade reaction times were significantly prolonged within one week post-injury in mTBI compared to controls. At follow-up assessment there was no significant between-groups difference. Changes in median SRT between the two assessments were not statistically significant. Four of the seven mTBI patients showed significantly increased SRT at follow-up; three of the mTBI patients and all controls showed no significant change. Among the three mTBI patients with persistent decreased SRT, two experienced loss of consciousness and reported the greatest symptoms, while the third was the only subject with significant decrease in neuropsychological testing scores at both assessments. CONCLUSION: In three of seven mTBI patients, saccadic eye movements remained delayed within three weeks post-injury. These three patients also showed persistent symptoms or no improvement on neuropsychological testing. This pilot study using a portable saccadometer suggests that comparing SRT from three weeks post-injury to that within one week of injury may be useful for early detection of a subpopulation at risk of persistent disability from mTBI. This finding suggests that further investigation in a large study population is warranted.Les saccades oculaires dans le traumatisme cérébral léger : une étude pilote.

Mania-like behavior induced by genetic dysfunction of the neuron-specific Na+,K+-ATPase α3 sodium pump.

Bipolar disorder is a debilitating psychopathology with unknown etiology. Accumulating evidence suggests the possible involvement of Na(+),K(+)-ATPase dysfunction in the pathophysiology of bipolar disorder. Here we show that Myshkin mice carrying an inactivating mutation in the neuron-specific Na(+),K(+)-ATPase α3 subunit display a behavioral profile remarkably similar to bipolar patients in the manic state. Myshkin mice show increased Ca(2+) signaling in cultured cortical neurons and phospho-activation of extracellular signal regulated kinase (ERK) and Akt in the hippocampus. The mood-stabilizing drugs lithium and valproic acid, specific ERK inhibitor SL327, rostafuroxin, and transgenic expression of a functional Na(+),K(+)-ATPase α3 protein rescue the mania-like phenotype of Myshkin mice. These findings establish Myshkin mice as a unique model of mania, reveal an important role for Na(+),K(+)-ATPase α3 in the control of mania-like behavior, and identify Na(+),K(+)-ATPase α3, its physiological regulators and downstream signal transduction pathways as putative targets for the design of new antimanic therapies.

Ciliary body lymphangiogenesis in uveal melanoma with and without extraocular extension.

PURPOSE: To determine whether peritumoral ciliary body lymphatics are found in uveal melanoma in the absence of extraocular extension. DESIGN: Consecutive case series from 1999 to 2005. PARTICIPANTS: Thirty-two uveal melanoma cases involving the ciliary body from the Ophthalmic Pathology Laboratory, University of Toronto, of which 23 showed no extraocular extension. METHODS: All immunofluorescence studies and quantitative analyses were performed in a masked fashion. Sections were immunostained for the presence of lymphatic endothelium using podoplanin (D2-40 antibody) and blood vessel endothelium using CD34. MAIN OUTCOME MEASURES: Identification and quantification of D2-40-positive lymphatic vessels in the ciliary body. RESULTS: In every case (n = 32), D2-40-positive lymphatics were detected in the peritumoral ciliary body. Lymphatic signal was significantly increased in the peritumoral ciliary body compared with the nonperitumoral ciliary body (P < 0.0001). There was no difference in lymphatic signal between cases with and without extraocular extension (P > 0.05). Lymphatics were not detected within the tumors. CONCLUSIONS: Peritumoral lymphangiogenesis was present in the ciliary body in uveal melanomas with and without extraocular extension, and as such, the presence of peritumoral lymphatics is not recommended as a prognostic marker in uveal melanoma.

Eye-brain axis in microgravity and its implications for Spaceflight Associated Neuro-ocular Syndrome.

Long-duration human spaceflight can lead to changes in both the eye and the brain, which have been referred to as Spaceflight Associated Neuro-ocular Syndrome (SANS). These changes may manifest as a constellation of symptoms, which can include optic disc edema, optic nerve sheath distension, choroidal folds, globe flattening, hyperopic shift, and cotton wool spots. Although the underpinning mechanisms for SANS are not yet known, contributors may include intracranial interstitial fluid accumulation following microgravity induced headward fluid shift. Development and validation of SANS countermeasures contribute to our understanding of etiology and accelerate new technology including exercise modalities, Lower Body Negative Pressure suits, venous thigh cuffs, and Impedance Threshold Devices. However, significant knowledge gaps remain including biomarkers, a full set of countermeasures and/or treatment regimes, and finally reliable ground based analogs to accelerate the research. This review from the European Space Agency SANS expert group summarizes past research and current knowledge on SANS, potential countermeasures, and key knowledge gaps, to further our understanding, prevention, and treatment of SANS both during human spaceflight and future extraterrestrial surface exploration.

Desialylated Platelet Clearance in the Liver is a Novel Mechanism of Systemic Immunosuppression.

Platelets are small, versatile blood cells that are critical for hemostasis/thrombosis. Local platelet accumulation is a known contributor to proinflammation in various disease states. However, the anti-inflammatory/immunosuppressive potential of platelets has been poorly explored. Here, we uncovered, unexpectedly, desialylated platelets (dPLTs) down-regulated immune responses against both platelet-associated and -independent antigen challenges. Utilizing multispectral photoacoustic tomography, we tracked dPLT trafficking to gut vasculature and an exclusive Kupffer cell-mediated dPLT clearance in the liver, a process that we identified to be synergistically dependent on platelet glycoprotein Ibα and hepatic Ashwell-Morell receptor. Mechanistically, Kupffer cell clearance of dPLT potentiated a systemic immunosuppressive state with increased anti-inflammatory cytokines and circulating CD4+ regulatory T cells, abolishable by Kupffer cell depletion. Last, in a clinically relevant model of hemophilia A, presensitization with dPLT attenuated anti-factor VIII antibody production after factor VIII ( infusion. As platelet desialylation commonly occurs in daily-aged and activated platelets, these findings open new avenues toward understanding immune homeostasis and potentiate the therapeutic potential of dPLT and engineered dPLT transfusions in controlling autoimmune and alloimmune diseases.

Intraocular extension of conjunctival invasive squamous cell carcinoma after pterygium surgery and cataract extraction.

OBJECTIVES: Conjunctival squamous dysplasia can often be confused with pterygium and pinguecula. Incomplete excision of dysplastic tissue can lead to recurrence and rarely intraocular invasion. This study describes two cases in which invasive squamous cell carcinoma (SCC) of the conjunctiva was originally partially resected as pterygium and eventually required enucleation for intraocular invasion. METHODS: In this clinicopathologic small case series, two cases of intraocular SCC managed at a single tertiary ocular oncology institution are described. Clinical features, pathologic characteristics, and relevant imaging are described. RESULTS: In both cases, incomplete excision of conjunctival SCC was followed by rapid regrowth of the conjunctival lesion and signs of intraocular inflammation. An intraocular mass within the substance of the ciliary body was identified using ultrasound biomicroscopy in both the cases. Enucleation was performed. Pathologic features were typical to SCC. CONCLUSIONS: Intraocular spread on conjunctival SCC occurs only rarely but tends to follow recurrence of the conjunctival lesion after attempted excision. Modes of invasion may include direct invasion through sclera, along the tract of the anterior ciliary vessels, or inoculation through intraocular surgery incision.

A model to measure lymphatic drainage from the eye.

Intraocular pressure (IOP) is the most important risk factor for glaucoma development and progression. Most anti-glaucoma treatments aim to lower IOP by enhancing aqueous humor drainage from the eye. Aqueous humor drainage occurs via well-characterized trabecular meshwork (TM) and uveoscleral (UVS) pathways, and recently described ciliary body lymphatics. The relative contribution of the lymphatic pathway to aqueous drainage is not known. We developed a sheep model to quantitatively assess lymphatic drainage along with TM and UVS outflows. This study describes that model and presents our initial findings. Following intracameral injection of (125)I-bovine serum albumin (BSA), lymph was continuously collected via cannulated cervical lymphatic vessels and the thoracic lymphatic duct over either a 3-h or 5-h time period. In the same animals, blood samples were collected from the right jugular vein every 15 min. Lymphatic and TM drainage were quantitatively assessed by measuring (125)I-BSA in lymph and plasma, respectively. Radioactive tracer levels were also measured in UVS and "other" ocular tissue, as well as periocular tissue harvested 3 and 5 h post-injection. Tracer recovered from UVS tissue was used to estimate UVS drainage. The amount of (125)I-BSA recovered from different fluid and tissue compartments was expressed as a percentage of total recovered tracer. Three hours after tracer injection, percentage of tracer recovered in lymph and plasma was 1.64% ± 0.89% and 68.86% ± 9.27%, respectively (n = 8). The percentage of tracer in UVS, other ocular and periocular tissues was 19.87% ± 5.59%, 4.30% ± 3.31% and 5.32% ± 2.46%, respectively. At 5 h (n = 2), lymphatic drainage was increased (6.40% and 4.96% vs. 1.64%). On the other hand, the percentage of tracer recovered from UVS and other ocular tissue had decreased, and the percentage from periocular tissue showed no change. Lymphatic drainage increased steadily over the 3 h post-injection period, while TM drainage increased rapidly - reaching a plateau at 30 min. This quantitative sheep model enables assessment of relative contributions of lymphatic drainage, TM and UVS outflows, and may help to better understand the effects of glaucoma agents on outflow pathways.

Quantum dots trace lymphatic drainage from the mouse eye.

Glaucoma is a leading cause of blindness in the world, often associated with elevated eye pressure. Currently, all glaucoma treatments aim to lower eye pressure by improving fluid exit from the eye. We recently reported the presence of lymphatics in the human eye. The lymphatic circulation is known to drain fluid from organ tissues and, as such, lymphatics may also play a role in draining fluid from the eye. We investigated whether lymphatic drainage from the eye is present in mice by visualizing the trajectory of quantum dots once injected into the eye. Whole-body hyperspectral fluorescence imaging was performed in 17 live mice. In vivo imaging was conducted prior to injection, and 5, 20, 40 and 70 min, and 2, 6 and 24 h after injection. A quantum dot signal was observed in the left neck region at 6 h after tracer injection into the eye. Examination of immunofluorescence-labelled sections using confocal microscopy showed the presence of a quantum dot signal in the left submandibular lymph node. This is the first direct evidence of lymphatic drainage from the mouse eye. The use of quantum dots to image this lymphatic pathway in vivo is a novel tool to stimulate new treatments to reduce eye pressure and prevent blindness from glaucoma.

A novel photoacoustic-fluorescent contrast agent for quantitative imaging of lymphatic drainage.

In vivo near-infrared (NIR) photoacoustic imaging (PAI) studies using novel contrast agents require validation, often via fluorescence imaging. Bioconjugation of NIR dyes to proteins is a versatile platform to obtain contrast agents for specific biomedical applications. Nonfluorescent NIR dyes with higher photostability present advantages for quantitative PAI, compared to most fluorescent NIR dyes. However, they don't provide a fluorescence signal required for fluorescence imaging. Here, we designed a hybrid PA-fluorescent contrast agent by conjugating albumin with a NIR nonfluorescent dye (QC-1) and a visible spectrum fluorescent dye, a BODIPY derivative. The new hybrid tracer QC-1/BSA/BODIPY (QBB) had a low minimum detectable concentration (2.5µM), a steep linear range (2.4-54.4 µM; slope 3.39 E -5), and high photostability. Tracer signal was measured in vivo using PAI to quantify its drainage from eye to the neck and its localization in the neck lymph node was validated with postmortem fluorescence imaging.

Identification of lymphatics in the ciliary body of the human eye: a novel "uveolymphatic" outflow pathway.

Impaired aqueous humor flow from the eye may lead to elevated intraocular pressure and glaucoma. Drainage of aqueous fluid from the eye occurs through established routes that include conventional outflow via the trabecular meshwork, and an unconventional or uveoscleral outflow pathway involving the ciliary body. Based on the assumption that the eye lacks a lymphatic circulation, the possible role of lymphatics in the less well defined uveoscleral pathway has been largely ignored. Advances in lymphatic research have identified specific lymphatic markers such as podoplanin, a transmembrane mucin-type glycoprotein, and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1). Lymphatic channels were identified in the human ciliary body using immunofluorescence with D2-40 antibody for podoplanin, and LYVE-1 antibody. In keeping with the criteria for lymphatic vessels in conjunctiva used as positive control, D2-40 and LYVE-1-positive lymphatic channels in the ciliary body had a distinct lumen, were negative for blood vessel endothelial cell marker CD34, and were surrounded by either discontinuous or no collagen IV-positive basement membrane. Cryo-immunogold electron microscopy confirmed the presence D2-40-immunoreactivity in lymphatic endothelium in the human ciliary body. Fluorescent nanospheres injected into the anterior chamber of the sheep eye were detected in LYVE-1-positive channels of the ciliary body 15, 30, and 45 min following injection. Four hours following intracameral injection, Iodine-125 radio-labeled human serum albumin injected into the sheep eye (n = 5) was drained preferentially into cervical, retropharyngeal, submandibular and preauricular lymph nodes in the head and neck region compared to reference popliteal lymph nodes (P < 0.05). These findings collectively indicate the presence of distinct lymphatic channels in the human ciliary body, and that fluid and solutes flow at least partially through this system. The discovery of a uveolymphatic pathway in the eye is novel and highly relevant to studies of glaucoma and other eye diseases.

Electron Microscopic Evidence of Photoreceptor Outer-segments in the Trabecular Meshwork in a Case of Schwartz-Matsuo Syndrome.

PURPOSE: To describe a case of photoreceptor outer segment glaucoma (Schwartz-Matsuo syndrome) with electron microscopic evidence of photoreceptor outer segments in the trabecular meshwork (TM). DESIGN: This is a clinicopathologic case report. PARTICIPANT: A 48-year-old Filipino man. METHODS: Specimens of aqueous humor and TM in a clinical case of Schwartz-Matsuo syndrome were examined by electron microscopy. MAIN OUTCOME MEASURES: Electron photomicroscopy. RESULTS: Electron microscopy showed evidence of retinal photoreceptor outer-segments in both an aqueous humor and a TM specimen. CONCLUSION: Schwartz-Matsuo syndrome is associated with the presence of photoreceptor outer segments in the TM.

Oxidative injury to blood vessels and glia of the pre-laminar optic nerve head in human glaucoma.

Glaucoma is a leading cause of irreversible world blindness. Oxidative damage and vascular injury have been implicated in the pathogenesis of this disease. The purpose of this study was to determine in human primary open angle glaucoma whether oxidative injury occurs in pre-laminar optic nerve blood vessels and glial cells. Following IRB approval, sections from post-mortem primary open angle glaucoma eyes (n=5) with mean age of 77 +/- 9 yrs (+/-SD) were compared to normal control eyes (n=4) with mean age 70 +/- 9 yrs (Eye Bank of Canada). Immunostaining with nitrotyrosine, a footprint for peroxynitrite-mediated injury, was performed and sections were double-labeled with markers for vascular endothelial cells, perivascular smooth muscle cells, and astrocytes with CD34, smooth muscle actin (SMA), and glial fibrillary acidic protein (GFAP), respectively. Immunostaining was captured in a masked fashion using confocal microscopy, and defined regions of interest for blood vessels and glial tissue. Intensity measurements of supra-threshold area in pixels as percent of the total number of pixels were calculated using ImageJ (NIH) and compared using two-tailed Mann-Whitney nonparametric tests between glaucoma and control groups. Colocalization coefficients with cell-specific markers were determined and compared with random coefficients of correlation. Increased nitrotyrosine immunoreactivity was observed in pre-laminar optic nerve head blood vessels of primary open angle glaucoma eyes compared to controls and this difference was statistically significant (1.35 +/- 1.11% [+/-SD] vs. 0.01 +/- 0.01%, P=0.016). NT-immunoreactivity was also increased in the glial tissue surrounding the pre-laminar optic nerve head in the glaucoma group and compared to controls, and this difference was statistically significant (18.37 +/-12.80% vs. 0.08 +/- 0.04%, P=0.016). Colocalization studies demonstrated nitrotyrosine staining in vascular endothelial and smooth muscle cells, in addition to astrocytes. Correlation coefficients for CD34, SMA, and GFAP were 0.37, 0.52, and 0.64, respectively. Oxidative injury is present in blood vessels and astrocytes in the pre-laminar optic nerve head in human primary open angle glaucoma. Peroxynitrite-mediated oxidative injury, whether primary or secondary, may contribute to the pathobiology of glaucoma disease.

Paying attention to the cerebrovascular system in glaucoma.

In glaucoma, damage to vision pathways in the brain may contribute to vision dysfunction and loss. A large part of the central visual system is located within a watershed area of increased susceptibility to ischemic injury. In addition to glaucomatous central visual neuron injury, glaucoma patients may suffer silent vascular insults from cerebral small blood vessel disease. We hypothesize that "double-hit" mechanisms of injury, such as glaucoma and comorbid cerebrovascular conditions, play a role in glaucomatous susceptibility and disease manifestations in young and aging populations at risk.

Retinal tau pathology in human glaucomas.

BACKGROUND: Tau protein is a microtubule-associated protein critical to neuron structure and integrity. The abnormal hyperphosphorylated tau protein AT8 disrupts microtubules, interferes with axonal transport, and is associated with neuron injury in neurodegenerative diseases such as Alzheimer's disease. The purpose of this study was to assess the presence of tau protein and abnormal tau protein AT8 in human glaucomas and to determine whether abnormal tau protein plays a role in glaucomatous neural degeneration. METHODS: Sections from 11 surgical eye specimens with glaucoma from elevated intraocular pressure causes and 10 age-matched control eye specimens were immunostained for normal tau protein (BT2) and hyperphosphorylated tau protein (AT8). Postmortem specimens with incidental open-angle glaucoma (n = 6) were compared with controls (n = 3). Measurements of immunofluorescence intensity in glaucoma retinas were compared with those in control retinas. Abnormal tau AT8 and parvalbumin, a horizontal cell-specific marker, were studied with double-immunofluorescence techniques to determine colocalization. RESULTS: In surgical glaucoma specimens, normal tau protein was decreased in both the optic nerve and retina compared with age-matched controls. Abnormal tau AT8 was evident within the posterior retina, predominantly at the outer border of the inner nuclear layer in surgical glaucoma specimens, and this was not observed in controls or incidental glaucoma cases. Quantitative immunofluorescence techniques demonstrated significantly increased abnormal tau AT8 in surgical glaucoma specimens compared with controls. Abnormal tau AT8 colocalized with parvalbumin in horizontal cells of the retina. INTERPRETATION: Abnormal tau AT8, a marker of injury in various neurological diseases, is present in human glaucomas with uncontrolled intraocular pressure. The finding of abnormal tau protein in retinal horizontal cells may relate to elevated intraocular pressure and (or) neural degeneration in glaucoma. Tau protein abnormality in glaucoma underscores shared pathways with other neurodegenerative diseases.

Wavelength optimization in the multispectral photoacoustic tomography of the lymphatic drainage in mice.

Multispectral photoacoustic tomography provides mapping of the tissue chromophore distributions using sets of tunable laser wavelengths. With the overall goal of studying the dynamics of cerebrospinal fluid in mice in vivo, our work aims to minimize the number of wavelengths to reduce scanning time, improve the temporal resolution, reduce the energy deposition and avoid the tracer photobleaching while maintaining high image quality. To select small sets of wavelengths we directly searched for the combinations of wavelengths providing the best and worst image quality in comparison with a reference image obtained using 131 closely spaced wavelengths between 680 and 940 nm in terms of the peak signal-to-noise ratio (PSNR). We have shown that using the PSNR optimization method, additional improvements could be achieved over the wavelength set selected using the method of the minimization of the extinction matrix condition number.