Plant vacuoles serve as the primary intracellular compartments for inorganic phosphate (Pi) storage. Passage of Pi across vacuolar membranes plays a critical role in buffering the cytoplasmic Pi level against fluctuations of external Pi and metabolic activities. Here we demonstrate that the SPX-MFS proteins, designated as PHOSPHATE TRANSPORTER 5 family (PHT5), also named Vacuolar Phosphate Transporter (VPT), function as vacuolar Pi transporters. Based on 31P-magnetic resonance spectroscopy analysis, Arabidopsis pht5;1 loss-of-function mutants accumulate less Pi and exhibit a lower vacuolar-to-cytoplasmic Pi ratio than controls. Conversely, overexpression of PHT5 leads to massive Pi sequestration into vacuoles and altered regulation of Pi starvation-responsive genes. Furthermore, we show that heterologous expression of the rice homologue OsSPX-MFS1 mediates Pi influx to yeast vacuoles. Our findings show that a group of Pi transporters in vacuolar membranes regulate cytoplasmic Pi homeostasis and are required for fitness and plant growth.
Genetic divergences among mammalian strains are presented phenotypically in various aspects of physical appearance such as body shape and facial features. Yet how genetic diversity is expressed in brain function still remains unclear. Functional connectivity has been shown to be a valuable approach in characterizing the relationship between brain functions and behaviors. Alterations in the brain default mode network (DMN) have been found in human neuropsychological disorders. In this study we selected the spontaneously hypertensive rat (SHR) and the Wistar Kyoto rat (WKY), two inbred rat strains with close genetic origins, to investigate variations in the DMN. Our results showed that the major DMN differences are the activities in hippocampal area and caudate putamen region. This may be correlated to the hyperactive behavior of the SHR strain. Advanced animal model studies on variations in the DMN may have potential to shed new light on translational medicine, especially with regard to neuropsychological disorders.
Ultrasound tissue characterization is crucial for the detection of tissue abnormalities. Since the statistics of the backscattered ultrasound signals strongly depend on density and spatial arrangement of local scatterers, appropriate modeling of the backscattered signals may be capable of providing unique physiological information on local tissue properties. Among various techniques, the Nakagami imaging, realized in a window-based estimation scheme, has a good performance in assessing different scatterer statistics in tissues. However, inconsistent m values have been reported in literature and obtained only from a local tissue region, abating the reliability of Nakagami imaging in tissue characterization. The discrepancies in m values in relevant literature may stem from the nonuniformity of the ultrasound image resolution, which is often neglected. We therefore hypothesized that window-based Nakagami m estimation was highly associated with the regional spatial resolution of ultrasound imaging. To test this hypothesis, our study investigated the effect of beamforming methods, including synthetic aperture (SA), coherent plane wave compounding (CPWC), multi-focusing (MF), and single-focusing (SF), on window-based m parameter estimation from the perspective of the resolution cell. The statistics of m parameter distribution as a function of imaging depth were characterized by their mean, variance, and skewness. The phantom with a low scatterer density (16 scatterers mm(-3)) had significantly lower m values compared to the ones with high scatterer densities (32 and 64 scatterers mm(-3)). Results from the homogeneous phantom with 64 scatterers mm(-3) showed that SA, MF, and CPWC had relatively uniform lateral resolutions compared to SF and thus relatively constant m estimates at different imaging depths. Our findings suggest that an ultrasound imaging regime exhibiting invariant spatial resolution throughout the entire imaging field of view would be the most appropriate for Nakagami imaging for tissue characterization.
This paper presents new albumin-shelled Gd-DTPA microbubbles (MBs) that can concurrently serve as a dual-modality contrast agent for ultrasound (US) imaging and magnetic resonance (MR) imaging to assist blood-brain barrier (BBB) opening and detect intracerebral hemorrhage (ICH) during focused ultrasound brain drug delivery. Perfluorocarbon-filled albumin-(Gd-DTPA) MBs were prepared with a mean diameter of 2320 nm and concentration of 2.903×10(9) MBs ml(-1) using albumin-(Gd-DTPA) and by sonication with perfluorocarbon (C(3)F(8)) gas. The albumin-(Gd-DTPA) MBs were then centrifuged and the procedure was repeated until the free Gd(3+) ions were eliminated (which were detected by the xylenol orange sodium salt solution). The albumin-(Gd-DTPA) MBs were also characterized and evaluated both in vitro and in vivo by US and MR imaging. Focused US was used with the albumin-(Gd-DTPA) MBs to induce disruption of the BBB in 18 rats. BBB disruption was confirmed with contrast-enhanced T(1)-weighted turbo-spin-echo sequence MR imaging. Heavy T(2)*-weighted 3D fast low-angle shot sequence MR imaging was used to detect ICH. In vitro US imaging experiments showed that albumin-(Gd-DTPA) MBs can significantly enhance the US contrast in T(1)-, T(2)- and T(2)*-weighted MR images. The r(1) and r(2) relaxivities for Gd-DTPA were 7.69 and 21.35 s(-1)mM(-1), respectively, indicating that the MBs represent a positive contrast agent in T(1)-weighted images. In vivo MR imaging experiments on 18 rats showed that focused US combined with albumin-(Gd-DTPA) MBs can be used to both induce disruption of the BBB and detect ICH. To compare the signal intensity change between pure BBB opening and BBB opening accompanying ICH, albumin-(Gd-DTPA) MB imaging can provide a ratio of 5.14 with significant difference (p = 0.026), whereas Gd-DTPA imaging only provides a ratio of 2.13 and without significant difference (p = 0.108). The results indicate that albumin-(Gd-DTPA) MBs have potential as a US/MR dual-modality contrast agent for BBB opening and differentiating focused-US-induced BBB opening from ICH, and can monitor the focused ultrasound brain drug delivery process.
The method allowed successful tissue targeting without causing overheating of the ribs. One main advantage of this approach is the accurate estimation of phase aberration caused by heterogeneously porous ribs and other interposed tissues. This strategy might prove useful to assess the effectiveness and safety of focused ultrasound thermal ablation prior to transrib treatment.
Focused ultrasound phased array systems have attracted increased attention for brain therapy applications. However, such systems currently lack a direct and real-time method to intraoperatively monitor ultrasound pressure distribution for securing treatment. This study proposes a dual-mode ultrasound phased array system design to support transmit/receive operations for concurrent ultrasound exposure and backscattered focal beam reconstruction through a spherically focused ultrasound array. A 256-channel ultrasound transmission system was used to transmit focused ultrasonic energy (full 256 channels), with an extended implementation of multiple-channel receiving function (up to 64 channels) using the same 256-channel ultrasound array. A coherent backscatter-received beam formation algorithm was implemented to map the point spread function (PSF) and focal beam distribution under a free-field/transcranial environment setup, with the backscattering generated from a strong scatterer (a point reflector or a microbubble-perfused tube) or a weakly scattered tissue-mimicking graphite phantom. Our results showed that PSF and focal beam can be successfully reconstructed and visualized in free-field conditions and can also be transcranially reconstructed following skull-induced aberration correction. In vivo experiments were conducted to demonstrate its capability to preoperatively and semiquantitatively map a focal beam to guide blood-brain barrier opening. The proposed system may have potential for real-time guidance of ultrasound brain intervention, and may facilitate the design of a dual-mode ultrasound phased array for brain therapeutic applications.
Sonogram-based temperature estimation and elastography have both shown promise as methods of monitoring focused ultrasound (FUS) treatments to induce thermal ablation in tissue. However, each method has important limitations. Temperature estimates based on echo delays become invalid when the relationship between sound speed and temperature is nonlinear, and are further complicated by thermal expansion and other changes in tissue. Elastography can track thermal lesion formation over a wider range of elasticity, but with low specificity and high noise. Furthermore, this method is poor at small lesion detection. This study proposes integrating the two estimates to improve the quality of monitoring FUS-induced thermal lesions. Our unified computational kernel is tested on three types of phantoms. Experiments with type I and type II phantoms were conducted to calibrate the thermal mapping and elastography methods, respectively. The optimal settings were then used in experiments with the type III phantom, which contains ex vivo swine liver tissue. Three different spatial-peak temporal-average intensities (I(spta); 35, 133 and 240 W cm(-2)) were delivered with a sonication time of 60 s. The new procedure can closely monitor heating while identifying the dimensions of the thermal lesion, and is significantly better at the latter task than either approach alone. This work may help improve the current clinical practice, which employs sonograms to guide the FUS-induced thermal ablation procedure.
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