Timed degradation of Mcl-1 controls mitotic cell death

Mitotic arrest can result in cell death through the process of apoptosis. We have shown by live-cell imaging that the ubiquitin-proteasome dependent proteolysis of the apoptotic regulator Mcl-1 under the control of the anaphase-promoting complex or cyclosome (APC/C) provides a timing mechanism that distinguishes prolonged mitotic arrest from normal mitosis

Adaptive windowing in contrast-enhanced intravascular ultrasound imaging

Intravascular ultrasound (IVUS) is one of the most commonly-used interventional imaging techniques and has seen recent innovations which attempt to characterize the risk posed by atherosclerotic plaques. One such development is the use of microbubble contrast agents to image vasa vasorum, fine vessels which supply oxygen and nutrients to the walls of coronary arteries and typically have diameters less than 200 µm. The degree of vasa vasorum neovascularization within plaques is positively correlated with plaque vulnerability. Having recently presented a prototype dual-frequency transducer for contrast agent-specific intravascular imaging, here we describe signal processing approaches based on minimum variance (MV) beamforming and the phase coherence factor (PCF) for improving the spatial resolution and contrast-to-tissue ratio (CTR) in IVUS imaging. These approaches are examined through simulations, phantom studies, ex vivo studies in porcine arteries, and in vivo studies in chicken embryos. In phantom studies, PCF processing improved CTR by a mean of 4.2 dB, while combined MV and PCF processing improved spatial resolution by 41.7%. Improvements of 2.2 dB in CTR and 37.2% in resolution were observed in vivo. Applying these processing strategies can enhance image quality in conventional B-mode IVUS or in contrast-enhanced IVUS, where signal-to-noise ratio is relatively low and resolution is at a premium

Pseudoelasticity at Large Strains in Au Nanocrystals [post-print]

© 2018 American Physical Society. Pseudoelasticity in metals is typically associated with phase transformations (e.g., shape memory alloys) but has recently been observed in sub-10 nm Ag nanocrystals that rapidly recovered their original shape after deformation to large strains. The discovery of pseudoelasticity in nanoscale metals dramatically changes the current understanding of the properties of solids at the smallest length scales, and the motion of atoms at surfaces. Yet, it remains unclear whether pseudoelasticity exists in different metals and nanocrystal sizes. The challenge of observing deformation at atomistic to nanometer length scales has prevented a clear mechanistic understanding of nanoscale pseudoelasticity, although surface diffusion and dislocation-mediated processes have been proposed. We further the understanding of pseudoelasticity in nanoscale metals by using a diamond anvil cell to compress colloidal Au nanocrystals under quasihydrostatic and nonhydrostatic pressure conditions. Nanocrystal structural changes are measured using optical spectroscopy and transmission electron microscopy and modeled using electrodynamic theory. We find that 3.9 nm Au nanocrystals exhibit pseudoelastic shape recovery after deformation to large uniaxial strains of up to 20%, which is equivalent to an ellipsoid with an aspect ratio of 2. Nanocrystal absorbance efficiency does not recover after deformation, which indicates that crystalline defects may be trapped in the nanocrystals after deformation

Atypical APC/C-dependent degradation of Mcl-1 provides an apoptotic timer during mitotic arrest

The initiation of apoptosis in response to the disruption of mitosis provides surveillance against chromosome instability. Here, we show that proteolytic destruction of the key regulator Mcl-1 during an extended mitosis requires the anaphase-promoting complex or cyclosome (APC/C) and is independent of another ubiquitin E3 ligase, SCF Using live-cell imaging, we show that the loss of Mcl-1 during mitosis is dependent on a D box motif found in other APC/C substrates, while an isoleucine-arginine (IR) C-terminal tail regulates the manner in which Mcl-1 engages with the APC/C, converting Mcl-1 from a Cdc20-dependent and checkpoint-controlled substrate to one that is degraded independently of checkpoint strength. This mechanism ensures a relatively slow but steady rate of Mcl-1 degradation during mitosis and avoids its catastrophic destruction when the mitotic checkpoint is satisfied, providing an apoptotic timer that can distinguish a prolonged mitotic delay from normal mitosis. Importantly, we also show that inhibition of Cdc20 promotes mitotic cell death more effectively than loss of APC/C activity through differential effects on Mcl-1 degradation, providing an improved strategy to kill cancer cells

Molecular Acoustic Angiography: A New Technique for High-resolution Superharmonic Ultrasound Molecular Imaging

Ultrasound molecular imaging utilizes targeted microbubbles to bind to vascular targets such as integrins, selectins, and other extracellular binding domains. After binding, these microbubbles are typically imaged using low pressures and multi-pulse imaging sequences. In this article, we present an alternative approach for molecular imaging using ultrasound which relies on superharmonic signals produced by microbubble contrast agents. Bound bubbles were insonified near resonance using a low frequency (4 MHz) and superharmonic echoes were received at high frequencies (25–30 MHz). While this approach was observed to produce declining image intensity during repeated imaging in both in vitro and in vivo experiments due to bubble destruction, the feasibility of superharmonic molecular imaging was demonstrated for transmit pressures which are sufficiently high to induce shell disruption in bound microbubbles. This approach was validated using microbubbles targeted to the αvβ3 integrin in a rat fibrosarcoma model (n=5), and combined with superharmonic images of free microbubbles to produce high contrast, high resolution 3D volumes of both microvascular anatomy and molecular targeting. Image intensity over repeated scans and the effect of microbubble diameter were also assessed in vivo, indicating that larger microbubbles yield increased persistence in image intensity. Using ultrasound-based acoustic angiography images rather than conventional B-mode ultrasound to provide the underlying anatomical information facilitates anatomical localization of molecular markers. Quantitative analysis of relationships between microvasculature and targeting information indicated that most targeting occurred within 50 µm of a resolvable vessel (>100 µm diameter). The combined information provided by these scans may present new opportunities for analyzing relationships between microvascular anatomy and vascular targets, subject only to limitations of the current mechanically-scanned system and microbubble persistence to repeated imaging at moderate mechanical indices

Ex Vivo Porcine Arterial and Chorioallantoic Membrane Acoustic Angiography Using Dual-Frequency Intravascular Ultrasound Probes

The presence of blood vessels within a developing atherosclerotic plaque has been shown to be correlated to increased plaque vulnerability and ensuing cardiac events, however, detecting coronary intraplaque neovascularizations poses a significant challenge in the clinic. In this paper, we demonstrate in vivo a new intravascular ultrasound imaging method using a dual-frequency transducer to visualize contrast flow in microvessels with high specificity. This method uses a specialized transducer capable of exciting contrast agents at a low frequency (5.5 MHz) while detecting their nonlinear superhamonics at a much higher frequency (37 MHz). In vitro evaluation of the approach was performed in a microvascular phantom to produce 3D renderings of simulated vessel patterns and to determine image quality metrics as a function of depth. Furthermore, the ability of the system to detect microvessels is demonstrated both ex vivo using porcine arteries and in vivo using the chorioallantoic membrane of a developing chicken embryo with optical confirmation. Dual-frequency contrast specific imaging was able to resolve vessels of a similar size to those found in vulnerable atherosclerotic plaques at clinically relevant depths. The results of this study adds growing support for further evaluation and translation of contrast specific imaging in intravascular ultrasound for the detection of vulnerable plaques in atherosclerosis

Electrical Anomalies Observed During DC3

The primary scientific goals of DC3 involved improving our understanding of the chemical impacts of thunderstorms and their anvils. However, the Colorado domain provided opportunities to study other interesting phenomena, including the potential impacts of smoke ingestion on convection and thunderstorms, electrification processes in smoke plumes and pyrocumulonimbus clouds, and the production of sprites by unconventional thunderstorm

Polarimetric and Multi-Doppler Radar Observations of Electrified and Unelectrified Wildfire Smoke Plumes

Pyrocumulus clouds above three Colorado wildfires (Hewlett Gulch, High Park, and Waldo Canyon; all occurred during summer 2012) electrified and produced small intracloud discharges whenever the smoke plumes grew to high altitudes (over 10 km above mean sea level, or MSL). This occurred during periods of rapid wildfire growth, as indicated by the shortwave infrared channel on a geostationary satellite, as well as by incident reports. In the Hewlett Gulch case, the fire growth led to increased updrafts within the plume, as inferred by multiple Doppler radar syntheses, which led to the vertical development and subsequent electrification a life cycle as short as 30 minutes. The lightning, detected by a threedimensional lightning mapping network, was favored in highaltitude regions (~10 km MSL) containing modest reflectivities (25 dBZ and lower), ~0 dB differential reflectivity, and reduced correlation coefficient (~0.60.7). This indicated the likely presence of ice particles (crystals and aggregates, possibly rimed) mixed with ash. Though neither multipleDoppler nor polarimetric observations were available during the electrification of the High Park and Waldo Canyon plumes, their NEXRAD observations showed reflectivity structures consistent with Hewlett Gulch. In addition, polarimetric and multipleDoppler scanning of unelectrified High Park plumes indicated only irregularly shaped ash, and not ice, was present (i.e., reflectivities 5 dB, correlation < 0.4), and there was no broaching of the 10 km altitude. Based on these results, the electrification likely was caused by icebased processes that did not involve significant amounts of graupel. The results demonstrate the scientific value of multipleDoppler and polarimetric radar observations of wildfire smoke plumes including the ability to distinguish between regions of pure hydrometeors, regions of pure ash, and mixtures of both and also suggest a possible new application for lightning data in monitoring wildfires

On the Electrification of Pyrocumulus Clouds

The electrification (or lack thereof) of pyrocumulus clouds is examined for several different wildfires that occurred during 20122013. For example, pyrocumulus clouds above three Colorado wildfires (Hewlett Gulch, High Park, and Waldo Canyon; all occurred during summer 2012) electrified and produced small intracloud discharges whenever the smoke plumes grew to high altitudes (over 10 km above mean sea level, or MSL). This occurred during periods of rapid wildfire growth, as indicated by the shortwave infrared channel on a geostationary satellite, as well as by incident reports. In the Hewlett Gulch case, the fire growth led to increased updrafts within the plume, as inferred by multipleDoppler radar syntheses, which led to the vertical development and subsequent electrification a life cycle as short as 30 minutes. The lightning, detected by a threedimensional lightning mapping network, was favored in highaltitude regions (~10 km MSL) containing modest reflectivities (25 dBZ and lower), ~0 dB differential reflectivity, and reduced correlation coefficient (~0.60.7). This indicated the likely presence of ice particles (crystals and aggregates, possibly rimed) mixed with ash. Though neither multipleDoppler nor polarimetric observations were available during the electrification of the High Park and Waldo Canyon plumes, their NEXRAD observations showed reflectivity structures consistent with Hewlett Gulch. In addition, polarimetric and multipleDoppler scanning of unelectrified High Park plumes indicated only irregularly shaped ash, and not ice, was present (i.e., reflectivities 5 dB, correlation < 0.4), and there was no broaching of the 10 km altitude. Based on these results, the electrification likely was caused by icebased processes that did not involve significant amounts of graupel. Results for pyrocumulus clouds above notable 2013 wildfires that also experienced rapid growth (e.g., Black Forest, Yarnell Hill, West Fork, Tres Lagunas, etc.) will be compared against the 2012 cases, with special emphasis on polarimetric NEXRAD and available lightning measurements, in order to better understand the physical processes responsible for pyrocumulus electrification