Thermal analysis of dry eye subjects and the thermal impulse perturbation model of ocular surface.

In this study, we explore the usage of ocular surface temperature (OST) decay patterns to distinguished between dry eye patients with aqueous deficient dry eye (ADDE) and meibomian gland dysfunction (MGD). The OST profiles of 20 dry eye subjects were measured by a long-wave infrared thermal camera in a standardized environment (24 °C, and relative humidity (RH) 40%). The subjects were instructed to blink every 5 s after 20 âˆ¼ 25 min acclimation. Exponential decay curves were fit to the average temperature within a region of the central cornea. We find the MGD subjects have both a higher initial temperature (p < 0.022) and a higher asymptotic temperature (p < 0.007) than the ADDE subjects. We hypothesize the temperature difference among the subpopulations is due to tear volume and heat transfer mechanisms. To study the validity of our claim, we develop a mathematical model, referred to as the thermal impulse perturbation (TIP) model. We conclude that long-wave-infrared thermal imaging is a plausible tool in assisting with the classification of dry eye patient.

Detection of microregional hypoxia in mouse cerebral cortex by two-photon imaging of endogenous NADH fluorescence.

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. Here we present a visualized experimental and methodological protocol to directly visualize microregional tissue hypoxia and to infer perivascular oxygen gradients in the mouse cortex. It is based on the non-linear relationship between nicotinamide adenine dinucleotide (NADH) endogenous fluorescence intensity and oxygen partial pressure in the tissue, where observed tissue NADH fluorescence abruptly increases at tissue oxygen levels below 10 mmHg(1). We use two-photon excitation at 740 nm which allows for concurrent excitation of intrinsic NADH tissue fluorescence and blood plasma contrasted with Texas-Red dextran. The advantages of this method over existing approaches include the following: it takes advantage of an intrinsic tissue signal and can be performed using standard two-photon in vivo imaging equipment; it permits continuous monitoring in the whole field of view with a depth resolution of ~50 µm. We demonstrate that brain tissue areas furthest from cerebral blood vessels correspond to vulnerable watershed areas which are the first to become functionally hypoxic following a decline in vascular oxygen supply. This method allows one to image microregional cortical oxygenation and is therefore useful for examining the role of inadequate or restricted tissue oxygen supply in neurovascular diseases and stroke.

Mitochondrial superoxide flashes: metabolic biomarkers of skeletal muscle activity and disease.

Mitochondrial superoxide flashes (mSOFs) are stochastic events of quantal mitochondrial superoxide generation. Here, we used flexor digitorum brevis muscle fibers from transgenic mice with muscle-specific expression of a novel mitochondrial-targeted superoxide biosensor (mt-cpYFP) to characterize mSOF activity in skeletal muscle at rest, following intense activity, and under pathological conditions. Results demonstrate that mSOF activity in muscle depended on electron transport chain and adenine nucleotide translocase functionality, but it was independent of cyclophilin-D-mediated mitochondrial permeability transition pore activity. The diverse spatial dimensions of individual mSOF events were found to reflect a complex underlying morphology of the mitochondrial network, as examined by electron microscopy. Muscle activity regulated mSOF activity in a biphasic manner. Specifically, mSOF frequency was significantly increased following brief tetanic stimulation (18.1 ± 1.6 to 22.3 ± 2.0 flashes/1000 µm²·100 s before and after 5 tetani) and markedly decreased (to 7.7 ± 1.6 flashes/1000 µm²·100 s) following prolonged tetanic stimulation (40 tetani). A significant temperature-dependent increase in mSOF frequency (11.9 ± 0.8 and 19.8 ± 2.6 flashes/1000 µm²·100 s at 23°C and 37°C) was observed in fibers from RYR1(Y522S/WT) mice, a mouse model of malignant hyperthermia and heat-induced hypermetabolism. Together, these results demonstrate that mSOF activity is a highly sensitive biomarker of mitochondrial respiration and the cellular metabolic state of muscle during physiological activity and pathological oxidative stress

Large-scale propagation of ultrasound in a 3-D breast model based on high-resolution MRI data.

A 40 x 35 x 25-mm(3) specimen of human breast consisting mostly of fat and connective tissue was imaged using a 3-T magnetic resonance scanner. The resolutions in the image plane and in the orthogonal direction were 130 microm and 150 microm, respectively. Initial processing to prepare the data for segmentation consisted of contrast inversion, interpolation, and noise reduction. Noise reduction used a multilevel bidirectional median filter to preserve edges. The volume of data was segmented into regions of fat and connective tissue by using a combination of local and global thresholding. Local thresholding was performed to preserve fine detail, while global thresholding was performed to minimize the interclass variance between voxels classified as background and voxels classified as object. After smoothing the data to avoid aliasing artifacts, the segmented data volume was visualized using isosurfaces. The isosurfaces were enhanced using transparency, lighting, shading, reflectance, and animation. Computations of pulse propagation through the model illustrate its utility for the study of ultrasound aberration. The results show the feasibility of using the described combination of methods to demonstrate tissue morphology in a form that provides insight about the way ultrasound beams are aberrated in three dimensions by tissue.