Non-invasive voiding assessment in conscious mice

OBJECTIVE: To review available options of assessing murine bladder function and to evaluate a non-invasive technique suitable for long-term recording. METHODS: We reviewed previously described methods to record rodent bladder function. We used modified metabolic cages to capture novel recording tracings of mouse micturition. We evaluated our method in a pilot study with female mice undergoing partial bladder outlet obstruction or sham operation, respectively; half of the partial obstruction and sham group received treatment with an S6K-inhibitor, targeting the mTOR pathway, which is known to be implicated in bladder response to obstruction. RESULTS: Our non-invasive method using continuous urine weight recording reliably detected changes in murine bladder function resulting from partial bladder outlet obstruction or treatment with S6K-inhibitor. We found obstruction as well as treatment with S6K-inhibitor to correlate with a hyperactive voiding pattern. CONCLUSIONS: While invasive methods to assess murine bladder function largely disturb bladder histology and intrinsically render post-cystometry gene expression analysis of questionable value, continuous urine weight recording is a reliable, inexpensive, and critically non-invasive method to assess murine bladder function, suitable for a long-term application

Compact 0.7 mJ/11 ns eye-safe erbium laser

We report on the development of a compact diode-end-pumped eye-safe (similar to 1.54 mu m) passively-cooled Er, Yb:glass laser. The design of this laser is facilitated by the use of a double-pass pumping scheme and a special ZrO2 diffuse reflector for a uniform pump distribution. In the free-running mode, this laser generates 8.2 mJ/3 ms pulses with a slope efficiency of 15%. Passive Q-switching is provided by saturable absorbers made of transparent glass-ceramics containing Co2+:gamma-Ga2O3 or Co2+:MgAl2O4 nanocrystals with a spinel structure. In the latter case, 0.7 mJ/10.5 ns pulses are generated corresponding to >60 kW peak power and good beam quality (M-2 = 1.4). The designed laser is suitable for portable range-finders

Plasma Membrane Integrity and Survival of Melanoma Cells After Nanosecond Laser Pulses

Circulating tumor cells (CTCs) photoacoustic detection systems can aid clinical decision-making in the treatment of cancer. Interaction of melanin within melanoma cells with nanosecond laser pulses generates photoacoustic waves that make its detection possible. This study aims at: (1) determining melanoma cell survival after laser pulses of 6 ns at λ = 355 and 532 nm; (2) comparing the potential enhancement in the photoacoustic signal using λ = 355 nm in contrast with λ = 532 nm; (3) determining the critical laser fluence at which melanin begins to leak out from melanoma cells; and (4) developing a time-resolved imaging (TRI) system to study the intracellular interactions and their effect on the plasma membrane integrity. Monolayers of melanoma cells were grown on tissue culture-treated clusters and irradiated with up to 1.0 J/cm2. Surviving cells were stained with trypan blue and counted using a hemacytometer. The phosphate buffered saline absorbance was measured with a nanodrop spectrophotometer to detect melanin leakage from the melanoma cells post-laser irradiation. Photoacoustic signal magnitude was studied at both wavelengths using piezoelectric sensors. TRI with 6 ns resolution was used to image plasma membrane damage. Cell survival decreased proportionally with increasing laser fluence for both wavelengths, although the decrease is more pronounced for 355 nm radiation than for 532 nm. It was found that melanin leaks from cells equally for both wavelengths. No significant difference in photoacoustic signal was found between wavelengths. TRI showed clear damage to plasma membrane due to laser-induced bubble formation

Structure and nonlinear optical properties of novel transparent glass-ceramics based on Co2+:ZnO nanocrystals

Transparent glass-ceramics (GCs) based on Co2+:ZnO nanocrystals (mean diameter, 11 nm) are synthesized on the basis of cobalt-doped glasses of the K2O–ZnO–Al2O3–SiO2 system. For these GCs, the absorption band related to the 4A2(4F) → 4T1(4F) transition of Co2+ ions in tetrahedral sites spans until ~1.73 μm. Saturation of the absorption is demonstrated at 1.54 μm, with a saturation fluence Fs = 0.8 ± 0.1 Jcm−2 (σGSA = 1.7 ± 0.2 × 10−19 cm2) and a recovery time of 890 ± 10 ns. Passive Q-switching of an Er,Yb:glass laser is realized with the synthesized GCs. This laser generated 0.37 mJ/100 ns pulses at 1.54 μm. The developed GCs are promising as saturable absorbers for 1.6–1.7 μm crystalline erbium lasers

Colorization and Automated Segmentation of Human T2 MR Brain Images for Characterization of Soft Tissues

Characterization of tissues like brain by using magnetic resonance (MR) images and colorization of the gray scale image has been reported in the literature, along with the advantages and drawbacks. Here, we present two independent methods; (i) a novel colorization method to underscore the variability in brain MR images, indicative of the underlying physical density of bio tissue, (ii) a segmentation method (both hard and soft segmentation) to characterize gray brain MR images. The segmented images are then transformed into color using the above-mentioned colorization method, yielding promising results for manual tracing. Our color transformation incorporates the voxel classification by matching the luminance of voxels of the source MR image and provided color image by measuring the distance between them. The segmentation method is based on single-phase clustering for 2D and 3D image segmentation with a new auto centroid selection method, which divides the image into three distinct regions (gray matter (GM), white matter (WM), and cerebrospinal fluid (CSF) using prior anatomical knowledge). Results have been successfully validated on human T2-weighted (T2) brain MR images. The proposed method can be potentially applied to gray-scale images from other imaging modalities, in bringing out additional diagnostic tissue information contained in the colorized image processing approach as described

Meneco, a Topology-Based Gap-Filling Tool Applicable to Degraded Genome-Wide Metabolic Networks

International audienceIncreasing amounts of sequence data are becoming available for a wide range of non-model organisms. Investigating and modelling the metabolic behaviour of those organisms is highly relevant to understand their biology and ecology. As sequences are often incomplete and poorly annotated, draft networks of their metabolism largely suffer from incompleteness. Appropriate gap-filling methods to identify and add missing reactions are therefore required to address this issue. However, current tools rely on phenotypic or taxonomic information, or are very sensitive to the stoichiometric balance of metabolic reactions, especially concerning the co-factors. This type of information is often not available or at least prone to errors for newly-explored organisms. Here we introduce Meneco, a tool dedicated to the topological gap-filling of genome-scale draft metabolic networks. Meneco reformulates gap-filling as a qualitative combinatorial optimization problem, omitting constraints raised by the stoichiometry of a metabolic network considered in other methods, and solves this problem using Answer Set Programming. Run on several artificial test sets gathering 10,800 degraded Escherichia coli networks Meneco was able to efficiently identify essential reactions missing in networks at high degradation rates, outperforming the stoichiometry-based tools in scalability. To demonstrate the utility of Meneco we applied it to two case studies. Its application to recent metabolic networks reconstructed for the brown algal model Ectocarpus siliculosus and an associated bacterium Candidatus Phaeomarinobacter ectocarpi revealed several candidate metabolic pathways for algal-bacterial interactions. Then Meneco was used to reconstruct, from transcriptomic and metabolomic data, the first metabolic network for the microalga Euglena mutabilis. These two case studies show that Meneco is a versatile tool to complete draft genome-scale metabolic networks produced from heterogeneous data, and to suggest relevant reactions that explain the metabolic capacity of a biological system

Radiance detection of non-scattering inclusions in turbid media

Detection of non-scattering domains (voids) is an area of active research in biomedical optics. To avoid complexities of image reconstruction algorithms and requirements of a priori knowledge of void locations inherent to diffuse optical tomography (DOT), it would be useful to establish specific experimental signatures of voids that would help identify and detect them by other means. To address this, we present a radiance-based spectro-angular mapping approach that identifies void locations in the angular domain and establishes their spectral features. Using water-filled capillaries in scattering Intralipid as a test platform, we demonstrate perturbations in the directional photon density distribution produced by individual voids

Dual-Agent Photodynamic Therapy with Optical Clearing Eradicates Pigmented Melanoma in Preclinical Tumor Models

Treatment using light-activated photosensitizers (photodynamic therapy, PDT) has shown limited efficacy in pigmented melanoma, mainly due to the poor penetration of light in this tissue. Here, an optical clearing agent (OCA) was applied topically to a cutaneous melanoma model in mice shortly before PDT to increase the effective treatment depth by reducing the light scattering. This was used together with cellular and vascular-PDT, or a combination of both. The effect on tumor growth was measured by longitudinal ultrasound/photoacoustic imaging in vivo and by immunohistology after sacrifice. In a separate dorsal window chamber tumor model, angiographic optical coherence tomography (OCT) generated 3D tissue microvascular images, enabling direct in vivo assessment of treatment response. The optical clearing had minimal therapeutic effect on the in control, non-pigmented cutaneous melanomas but a statistically significant effect (p < 0.05) in pigmented lesions for both single- and dual-photosensitizer treatment regimes. The latter enabled full-depth eradication of tumor tissue, demonstrated by the absence of S100 and Ki67 immunostaining. These studies are the first to demonstrate complete melanoma response to PDT in an immunocompromised model in vivo, with quantitative assessment of tumor volume and thickness, confirmed by (immuno) histological analyses, and with non-pigmented melanomas used as controls to clarify the critical role of melanin in the PDT response. The results indicate the potential of OCA-enhanced PDT for the treatment of pigmented lesions, including melanoma

Changes in optical properties of ex vivo rat prostate due to heating

This study examines the effectiveness of a single, first-order Arrhenius process in accurately modelling the thermally induced changes in the optical properties, particularly the reduced scattering coefficient, μ'(s), and the absorption coefficient, μ(a), of ex vivo rat prostate. Recent work has shown that μ'(s) can increase as much as five-fold due to thermal coagulation, and the observed change in μ'(s) has been modelled well according to a first- order rate process in albumen. Conversely, optical property measurements conducted using pig liver suggest that this change in μ'(s) cannot suitably be described using a single rate parameter. In canine prostate, measurements have indicated that while the absorption coefficient varies with temperature, it does not do so according to first-order kinetics. A double integrating sphere system was used to measure the reflectance and transmittance of light at 810 nm through a thin sample of prostate. Using prostate samples collected from Sprague-Dawley rats, optical properties were measured at a constant elevated temperature. Tissue samples were measured over the range 54-83 °C. The optical properties of the sample were determined through comparison with reflectance and transmittance values predicted by a Monte Carlo simulation of light propagation in turbid media. A first-order Arrhenius model was applied to the observed change in μ'(s) and μ(a) to determine the rate process parameters for thermal coagulation. The measured rate coefficients were E(a) = (7.18 ± 1.74) x 104 J mol-1 and A(freq) = 3.14 x 108 s-1 for μ'(s). It was determined that the change in μ'(s) is well described by a single first-order rate process. Similar analysis performed on the changes in μ(a) due to increased temperatures yielded E(a) = (1.01 ± 0.35) x 105 J mol-1 and A(freq) = 8.92 x 1012 s-1. The results for μ(a) suggest that the Arrhenius model may be applicable to the changes in absorption