Dynamic Alterations in Blood Flow in Glaucoma Measured with Laser Speckle Contrast Imaging.

PURPOSE: To assess the repeatability of blood flow velocity index (BFVi) metrics obtained with a recently Food and Drug Administration-cleared laser speckle contrast imaging device, the XyCAM RI (Vasoptic Medical, Inc), and to characterize differences in these metrics among control, glaucoma suspect, and glaucoma participants. DESIGN: Prospective, observational study. PARTICIPANTS: Forty-six participants: 20 control, 16 glaucoma suspect, and 10 glaucoma participants, 1 eye per participant. METHODS: Key dynamic BFVi metrics-mean, peak, dip, volumetric rise index (VRI), volumetric fall index (VFI), time to rise (TtR), time to fall (TtF), blow-out time (BOT), skew, and acceleration time index-were measured in the optic disc, optic disc vessels, optic disc perfusion region, and macula in 4 imaging sessions on the same day. Intrasession and intersession variability were calculated using the coefficient of variation (CV) for each metric in each region of interest (ROI). Values for each dynamic BFVi variable were compared between glaucoma, glaucoma suspect, and control participants using bivariate and multivariate analysis. Pearson correlation coefficients were used to correlate each variable in each ROI with age, intraocular pressure, cup-to-disc ratio (CDR), mean deviation, pattern standard deviation, retinal nerve fiber layer thickness, and minimum rim width. MAIN OUTCOME MEASURES: Coefficient of variation for the intrasession and intersession variability for each dynamic BFVi metric in each ROI and differences in each metric in each ROI between each diagnostic group. RESULTS: Intersession CV for mean, peak, dip, VRI, VFI, TtR, and TtF ranged from 3.2 ± 2.5% to 11.0 ± 3.8%. Age, CDR, OCT metrics, and visual field metrics showed significant correlations with dynamic BFVi variables. Peak, mean, dip, VRI, and VFI were significantly lower in patients with glaucoma than in control participants in all ROIs except the fovea. These metrics also were significantly lower in glaucoma patients than glaucoma suspect patients in the disc vessels. CONCLUSIONS: Dynamic blood flow metrics measured with the XyCAM RI are reliable, are associated with structural and functional glaucoma metrics, and are significantly different among glaucoma, glaucoma suspect, and control participants. The XyCAM RI may serve as an important tool in glaucoma management in the future.

Portable, non-invasive video imaging of retinal blood flow dynamics.

Retinal blood flow (RBF) information has the potential to offer insight into ophthalmic health and disease that is complementary to traditional anatomical biomarkers as well as to retinal perfusion information provided by fluorescence or optical coherence tomography angiography (OCT-A). The present study was performed to test the functional attributes and performance of the XyCAM RI, a non-invasive imager that obtains and assesses RBF information. The XyCAM RI was installed and used in two different settings to obtain video recordings of the blood flow in the optic nerve head region in eyes of healthy subjects. The mean blood flow velocity index (BFVi) in the optic disc and in each of multiple arterial and venous segments was obtained and shown to reveal a temporal waveform with a peak and trough that correlates with a cardiac cycle as revealed by a reference pulse oximeter (correlation between respective peak-to-peak distances was 0.977). The intra-session repeatability of the XyCAM RI was high with a coefficient of variation (CV) of 1.84 ± 1.13% across both sites. Artery-vein comparisons were made by estimating, in a pair of adjacent arterial and venous segments, various temporal waveform metrics such as pulsatility index, percent time in systole and diastole, and change in vascular blood volume over a cardiac cycle. All arterial metrics were shown to have significant differences with venous metrics (p < 0.001). The XyCAM RI, therefore, by obtaining repeatable blood flow measurements with high temporal resolution, permits the differential assessment of arterial and venous blood flow patterns in the retina that may facilitate research into disease pathophysiology and biomarker development for diagnostics.

Global transcriptional downregulation of TREX and nuclear trafficking machinery as pan-senescence phenomena: evidence from human cells and tissues.

Nucleocytoplasmic trafficking (NCT) of macromolecules is a fundamental process in eukaryotes that requires tight controls to maintain proper cell functions. Downregulation of the classical NCT pathway in senescent cells has been reported. However, whether this is a hallmark that exists across all types of cellular senescence remains unknown, and whether the mRNA export machinery is altered during senescence has not been demonstrated. Here, we show that the global transcriptomic downregulation of both the TREX (transcription-export) machinery and classical NLS-dependent protein transport machinery is a hallmark of varying types of senescence. A gene set-based approach using 25 different studies showed that the TREX-NCT gene set displays distinct common downregulated patterns in senescent cells versus its expression in their nonsenescent counterparts regardless of the senescence type, such as replicative senescence (RS), tumor cell senescence (TCS), oncogene-induced senescence (OIS), stem cell senescence (SCS), progeria and endothelial cell senescence (ECS). Similar patterns of TREX-NCT gene downregulation were also shown in two large human tissue genomic databases, the Genotype-Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA) databases. We also found that early-stage cancer tissues show consistent age-related patterns of TREX-NCT enrichment, suggesting the potential significance of TREX-NCT genes in determining cell fate in the early stage of tumorigenesis. Moreover, human cancer tissues exhibit an opposite TREX-NCT enrichment pattern with aging, indicating that deviation from age-related changes in TREX-NCT genes may provide a novel but critical clue for the age-dependent pathogenesis of cancer and increase in cancer incidence with aging.

Variability in locomotor dynamics reveals the critical role of feedback in task control.

Animals vary considerably in size, shape, and physiological features across individuals, but yet achieve remarkably similar behavioral performances. We examined how animals compensate for morphophysiological variation by measuring the system dynamics of individual knifefish (Eigenmannia virescens) in a refuge tracking task. Kinematic measurements of Eigenmannia were used to generate individualized estimates of each fish's locomotor plant and controller, revealing substantial variability between fish. To test the impact of this variability on behavioral performance, these models were used to perform simulated 'brain transplants'-computationally swapping controllers and plants between individuals. We found that simulated closed-loop performance was robust to mismatch between plant and controller. This suggests that animals rely on feedback rather than precisely tuned neural controllers to compensate for morphophysiological variability.

People come in different shapes and sizes, but most will perform similarly well if asked to complete a task requiring fine manual dexterity ­ such as holding a pen or picking up a single grape. How can different individuals, with different sized hands and muscles, produce such similar movements? One explanation is that an individual's brain and nervous system become precisely tuned to mechanics of the body's muscles and skeleton. An alternative explanation is that brain and nervous system use a more "robust" control policy that can compensate for differences in the body by relying on feedback from the senses to guide the movements. To distinguish between these two explanations, Uyanik et al. turned to weakly electric freshwater fish known as glass knifefish. These fish seek refuge within root systems, reed grass and among other objects in the water. They swim backwards and forwards to stay hidden despite constantly changing currents. Each fish shuttles back and forth by moving a long ribbon-like fin on the underside of its body. Uyanik et al. measured the movements of the ribbon fin under controlled conditions in the laboratory, and then used the data to create computer models of the brain and body of each fish. The models of each fish's brain and body were quite different. To study how the brain interacts with the body, Uyanik et al. then conducted experiments reminiscent of those described in the story of Frankenstein and transplanted the brain from each computer model into the body of different model fish. These "brain swaps" had almost no effect on the model's simulated swimming behavior. Instead, these "Frankenfish" used sensory feedback to compensate for any mismatch between their brain and body. This suggests that, for some behaviors, an animal's brain does not need to be precisely tuned to the specific characteristics of its body. Instead, robust control of movement relies on many seemingly redundant systems that provide sensory feedback. This has implications for the field of robotics. It further suggests that when designing robots, engineers should prioritize enabling the robots to use sensory feedback to cope with unexpected events, a well-known idea in control engineering.

Down-regulation of receptor-mediated endocytosis is responsible for senescence-associated hyporesponsiveness.

Human diploid fibroblasts (HDF) do not divide indefinitely and eventually lead to an arrest of cell division by a process termed cellular or replicative senescence. Irreversible growth arrest of senescent cells is strongly related to the attenuated response to growth factors. Recently, we reported that up-regulation of caveolin in the senescent cells is responsible for the attenuated response to growth factors. Senescent cells did not phosphorylate Erk-1/2 after EGF stimulation, whereas young cells did. In those senescent cells, we found an increased level of caveolin proteins and strong interactions between caveolin-1 and EGFR. When we overexpressed caveolin-1 in young HDF, the activation of Erk-1/2 on EGF stimulation was significantly suppressed. These results suggest that the hyporesponsiveness of senescent fibroblasts to EGF stimulation might be due to the overexpression of caveolin. In addition, the clathrin-dependent endocytosis system plays the more active and dominant role over the caveolae system. Therefore, we monitored the efficiency of clathrin-dependent receptor-mediated endocytosis in the senescent cells in order to elucidate the exact mode of the attenuated response to growth factors in the senescent cells. Using a transferrin-uptake assay and Western blot analysis of endocytosis-related proteins, we found a significant decrease of amphiphysin-1 in human diploid fibroblasts of multipassages. By adjusting the level of amphiphysin, we could modulate the efficiency of receptor-mediated endocytosis either in young or old cells toward growth factors: that is, a dominant negative mutant of amphiphysin-1 blocked the endocytosis in the young cells, while microinjection of the gene resumed its activity in the old cells. Taken together, we conclude that the loss of endocytotic activity of senescent cells is directly related to the down-regulation of amphiphysin-1 and/or up-regulation of caveolins. This opens a new field of functional recovery of the senescent cells simply through adjusting the receptor-mediated endocytosis capacity.