Dynamic Views of Structure and Function during Heart Morphogenesis

Congenital heart defects remain the most common birth defect in humans, occurring in over 1% of live births. The high prevalence of cardiac malformations can be partially attributed to limited knowledge regarding the embryonic roots of the disease. A variety of congenital heart defects are thought to arise from combinations of genetic and epigenetic factors. In an effort to better understand this dynamic relationship, our study explores the structure and function of the developing heart and valves and examines hemodynamic factors influencing valvulogenesis. In order to study cardiac mechanics, we employed novel high-speed confocal microscopy and four-dimensional visualization techniques. A dynamic four-dimensional dataset describing heart and valve development along with blood flow patterns throughout cardiac morphogenesis is presented. Utilizing newly developed tools, we propose a novel pumping mechanism in the valveless embryonic heart tube via elastic wave propagation and reflection. We show that this form of pumping leads to oscillatory shear stresses in the developing atrio-ventricular canal, a phenomenon that had not previously been documented. An in vivo method to modulate trans-valvular oscillatory flows is described and used to test our hypothesis that oscillatory shear stress across the primitive valve cushions stimulates heart valve leaflet formation. Our results suggest hemodynamic forces contribute to valvulogenesis and enhance our understanding of normal and abnormal heart valve development

Strained layer InP/InGaAs quantum well laser

Strained layer single or multiple quantum well lasers include an InP substrate, a pair of lattice-matched InGaAsP quarternary layers epitaxially grown on the substrate surrounding a pair of lattice matched In.sub.0.53 Ga.sub.0.47 As ternary layers surrounding one or more strained active layers of epitaxially grown, lattice-mismatched In.sub.0.75 Ga.sub.0.25 As. The level of strain is selected to control the bandgap energy to produce laser output having a wavelength in the range of 1.6 to 2.5 .mu.m. The multiple quantum well structure uses between each active layer. Diethyl zinc is used for p-type dopant in an InP cladding layer at a concentration level in the range of about 5.times.10.sup.17 /cm.sup.3 to about 2.times.10.sup.18 /cm.sup.3. Hydrogen sulfide is used for n-type dopant in the substrate

Index-Coupled Distributed-Feedback Semiconductor Quantum Cascade Lasers Fabricated Without Epitaxial Regrowth

Quantum cascade (QC) lasers and methods of fabricating such QC lasers are provided. The QC lasers incorporate a DFB grating without requiring the use of relying on epitaxial regrowth processes. The DFB gratings are formed as sidewall gratings along the lateral length of the QC active region, or the DFB gratings are formed atop the lateral length of the QC active region, and wherein the top DFB grating is planarized with a polymeric material

Acute toxicity effect of glyphosate on survival rate of common carp, Cyprinus carpio

Background: Herbicides are usually used to control weeds and some of them like glyphosate are nonselective herbicides. Aquatic environments are usually the last destinations of agricultural pesticides, which disrupt the metabolic processes of organisms. Thus, the aim of this study was to evaluate acute toxicity of glyphosate on the survival rate of common carp, Cyprinus carpio. Methods: A total of 135 common carp averaged 7 ± 0.8 g in weight were exposed to 0, 25, 50, 75, 100, 125, 150, 175 and 200 mL.L-1 glyphosate (15 fish in each treatment, with triplicates) for 96 hours. The aquariums capacity was 98 L in volume (80 × 35 × 35 cm) and physicochemical parameters were the same for all groups (pH 7.4-8, temperature = 26 ± 1ºC, DO = 7 mg.L-1 and total hardness of 190 mg CaCO3). LC10, LC20, LC30, LC40, LC50, LC60, LC70, LC80, LC90 and LC95 of glyphosate were calculated at 24, 48, 72 and 96 hours after adding glyphosate using probit test. Results: Mortality was observed in all treatments which exposed to higher than 50 mL.L-1 after 96 hours. The results showed that 96-hour LC50 of glyphosate for common carp was 92.71 mL.L-1. The fish exposed to different concentrations of glyphosate showed clinical signs such as increased mucus secretion, skin darkening and death with mouth open. Conclusion: Glyphosate disrupts the synthesis of amino acids in plants by inhibiting enzymatic activity of 5-enolpyruvylshikimate-3-phosphate synthase (EPSP). This enzyme is absent in animals. However, glyphosate is toxic for common carp. Keywords: Animals, Herbicides, Glyphosate, Carps, Excitatory postsynaptic potential

Four-dimensional cardiac imaging in living embryos via postacquisition synchronization of nongated slice sequences

Being able to acquire, visualize, and analyze 3D time series (4D data) from living embryos makes it possible to understand complex dynamic movements at early stages of embryonic development. Despite recent technological breakthroughs in 2D dynamic imaging, confocal microscopes remain quite slow at capturing optical sections at successive depths. However, when the studied motion is periodic— such as for a beating heart—a way to circumvent this problem is to acquire, successively, sets of 2D+time slice sequences at increasing depths over at least one time period and later rearrange them to recover a 3D+time sequence. In other imaging modalities at macroscopic scales, external gating signals, e.g., an electro-cardiogram, have been used to achieve proper synchronization. Since gating signals are either unavailable or cumbersome to acquire in microscopic organisms, we have developed a procedure to reconstruct volumes based solely on the information contained in the image sequences. The central part of the algorithm is a least-squares minimization of an objective criterion that depends on the similarity between the data from neighboring depths. Owing to a wavelet-based multiresolution approach, our method is robust to common confocal microscopy artifacts. We validate the procedure on both simulated data and in vivo measurements from living zebrafish embryos

Structure and function relationship of Zebrafish embryonic heart from confocal microscopy images

Confocal microscopy enables us to track myocytes in the embryonic zebrafish heart. The Zeiss LSM 5 Live high speed confocal microscope has been used to take optical sections (at 3 µm intervals and 151 frames per second) through a fluorescently labeled zebrafish heart at two developmental stages (26 and 34 hours post fertilization (hpf)). This data provides unique information allowing us to conjecture on the morphology and biomechanics of the developing vertebrate heart. Nevertheless, the myocytes, whose positions could be determined in a reliable manner, were located sparsely and mostly in one side of the heart tube. This difficulty was overcome using computational methods, that give longitudinal, radial and circumferential displacements of the myocytes as well as their contractile behavior. Applied strain analysis has shown that in the early embryonic heart tube, only the caudal region (near the in-flow) and another point in the middle of the tube can be active; the rest appears to be mostly passive. This statement is based on the delay between major strain and displacement which a material point experiences. Wave-like propagation of all three components of the displacement, especially in the circumferential direction, as well as the almost-periodic changes of the maximum strain support the hypothesis of helical muscle structure embedded in the tube. Changes of geometry in the embryonic heart after several hours are used to verify speculations about the structure based on the earlier images and aforementioned methods

Nonuniform temporal alignment of slice sequences for four-dimensional imaging of cyclically deforming embryonic structures

The temporal alignment of nongated slice-sequences acquired at different axial positions in the living embryonic zebrafish heart permits the reconstruction of dynamic, three-dimensional data. This approach overcomes the current acquisition- speed limitation of confocal microscopes for real-time three-dimensional imaging of fast processes. Current synchronization methods align and uniformly scale the data in time, but do not compensate for slight variations in the heart rhythm that occur within a heartbeat. Therefore, they impose constraints on the admissible data quality. Here, we derive a nonuniform registration procedure based on the minimization of the absolute value of the intensity difference between adjacent slice-sequence pairs. The method compensates for temporal intra-sample variations and allows the processing of a wider range of data to build functional, dynamic models of the beating embryonic heart. We show reconstructions from data acquired in living, fluorescent zebrafish embryos

Miniature Laser Doppler Velocimeter for Measuring Wall Shear

A miniature optoelectronic instrument has been invented as a nonintrusive means of measuring a velocity gradient proportional to a shear stress in a flow near a wall. The instrument, which can be mounted flush with the wall, is a variant of a basic laser Doppler velocimeter. The laser Doppler probe volume can be located close enough to the wall (as little as 100 micron from the surface) to lie within the viscosity-dominated sublayer of a turbulent boundary layer. The instrument includes a diode laser, the output of which is shaped by a diffractive optical element (DOE) into two beams that have elliptical cross sections with very high aspect ratios

High-Power Single-Mode 2.65-micron InGaAsSb/AlInGaAsSb Diode Lasers

Central to the advancement of both satellite and in-situ science are improvements in continuous-wave and pulsed infrared laser systems coupled with integrated miniaturized optics and electronics, allowing for the use of powerful, single-mode light sources aboard both satellite and unmanned aerial vehicle platforms. There is a technological gap in supplying adequate laser sources to address the mid-infrared spectral window for spectroscopic characterization of important atmospheric gases. For high-power applications between 2 to 3 micron, commercial laser technologies are unsuitable because of limitations in output power. For instance, existing InP-based laser systems developed for fiber-based telecommunications cannot be extended to wavelengths longer than 2 micron. For emission wavelengths shorter than 3 micron, intersubband devices, such as infrared quantum cascade lasers, become inefficient due to band-offset limitations. To date, successfully demonstrated singlemode GaSb-based laser diodes emitting between 2 and 3 micron have employed lossy metal Bragg gratings for distributed- feedback coupling, which limits output power due to optical absorption. By optimizing both the quantum well design and the grating fabrication process, index-coupled distributed-feedback 2.65-micron lasers capable of emitting in excess of 25 mW at room temperature have been demonstrated. Specifically, lasers at 3,777/cm (2.65 micron) have been realized to interact with strong absorption lines of HDO and other isotopologues of H2O. With minor modifications of the optical cavity and quantum well designs, lasers can be fabricated at any wavelength within the 2-to-3-micron spectral window with similar performance. At the time of this reporting, lasers with this output power and wavelength accuracy are not commercially available. Monolithic ridge-waveguide GaSb lasers were fabricated that utilize secondorder lateral Bragg gratings to generate single-mode emission from InGaAsSb/ AlInGaAsSb multi-quantum well structures. The device fabrication utilizes etched index-coupled gratings in the top AlGaAsSb cladding of the laser chip along the ridge waveguide, whereas commercial lasers that emit close to this wavelength include loss-coupled metal gratings that limit the output power of the laser. Semiconductor-laser-based spectrometers can be used to replace gas sensors currently used in industry and government. With the availability of high-power laser sources at mid-infrared wavelengths, sensors can target strong fundamental gas absorption lines to maximize instrument sensitivity

Quantitative mapping of intracardiac blood flow in embryonic zebrafish

Using real-time in vivo imaging and digital particle image velocimetry (DPIV) we quantitatively described the intracardiac flow environment of early zebrafish (Danio rerio) embryos. Gross cardiac dynamics were defined for two embryonic stages: 4.5 days post fertilization (dpf) and 37 hours post fertilization (hpf) using high-speed transmitted light microscopy with valve dynamics visualized through high-speed laser-scanning microscopy on transgenic embryos expressing GFP