In Vivo Imaging of the Buccal Mucosa Shows Loss of the Endothelial Glycocalyx and Perivascular Hemorrhages in Pediatric Plasmodium falciparum Malaria.

Severe malaria is mostly caused by Plasmodium falciparum, resulting in considerable, systemic inflammation and pronounced endothelial activation. The endothelium forms an interface between blood and tissue, and vasculopathy has previously been linked with malaria severity. We studied the extent to which the endothelial glycocalyx that normally maintains endothelial function is involved in falciparum malaria pathogenesis by using incident dark-field imaging in the buccal mucosa. This enabled calculation of the perfused boundary region, which indicates to what extent erythrocytes can permeate the endothelial glycocalyx. The perfused boundary region was significantly increased in severe malaria patients and mirrored by an increase of soluble glycocalyx components in plasma. This is suggestive of a substantial endothelial glycocalyx loss. Patients with severe malaria had significantly higher plasma levels of sulfated glycosaminoglycans than patients with uncomplicated malaria, whereas other measured glycocalyx markers were raised to a comparable extent in both groups. In severe malaria, the plasma level of the glycosaminoglycan hyaluronic acid was positively correlated with the perfused boundary region in the buccal cavity. Plasma hyaluronic acid and heparan sulfate were particularly high in severe malaria patients with a low Blantyre coma score, suggesting involvement in its pathogenesis. In vivo imaging also detected perivascular hemorrhages and sequestering late-stage parasites. In line with this, plasma angiopoietin-1 was decreased while angiopoietin-2 was increased, suggesting vascular instability. The density of hemorrhages correlated negatively with plasma levels of angiopoietin-1. Our findings indicate that as with experimental malaria, the loss of endothelial glycocalyx is associated with vascular dysfunction in human malaria and is related to severity.

Universal Synthetic Aperture Sequence for Anatomic, Functional, and Super Resolution Imaging.

Synthetic aperture (SA) can be used for both anatomic and functional imaging, where tissue motion and blood velocity are revealed. Often, sequences optimized for anatomic B-mode imaging are different from functional sequences, as the best distribution and number of emissions are different. B-mode sequences demand many emissions for a high contrast, whereas flow sequences demand short sequences for high correlations yielding accurate velocity estimates. This article hypothesizes that a single, universal sequence can be developed for linear array SA imaging. This sequence yields high-quality linear and nonlinear B-mode images as well as accurate motion and flow estimates for high and low blood velocities and super-resolution images. Interleaved sequences with positive and negative pulse emissions for the same spherical virtual source were used to enable flow estimation for high velocities and make continuous long acquisitions for low-velocity estimation. An optimized pulse inversion (PI) sequence with 2 ×12 virtual sources was implemented for four different linear array probes connected to either a Verasonics Vantage 256 scanner or the SARUS experimental scanner. The virtual sources were evenly distributed over the whole aperture and permuted in emission order for making flow estimation possible using 4, 8, or 12 virtual sources. The frame rate was 208 Hz for fully independent images for a pulse repetition frequency of 5 kHz, and recursive imaging yielded 5000 images per second. Data were acquired from a phantom mimicking the carotid artery with pulsating flow and the kidney of a Sprague-Dawley rat. Examples include anatomic high contrast B-mode, non-linear B-mode, tissue motion, power Doppler, color flow mapping (CFM), vector velocity imaging, and super-resolution imaging (SRI) derived from the same dataset and demonstrate that all imaging modes can be shown retrospectively and quantitative data derived from it.

Precise Estimation of Intravascular Pressure Gradients.

This study presents a method for noninvasive pressure gradient estimation, which allows the detection of small pressure differences with higher precision compared to invasive catheters. It combines a new method for estimating the temporal acceleration of the flowing blood with the Navier-Stokes equation. The acceleration estimation is based on a double cross-correlation approach, which is hypothesized to minimize the influence of noise. Data are acquired using a 256-element, 6.5-MHz GE L3-12-D linear array transducer connected to a Verasonics research scanner. A synthetic aperture (SA) interleaved sequence with 2 ×12 virtual sources evenly distributed over the aperture and permuted in emission order is used in combination with recursive imaging. This enables a temporal resolution between correlation frames equal to the pulse repetition time at a frame rate of half the pulse repetition frequency. The accuracy of the method is evaluated against a computational fluid dynamic simulation. Here, the estimated total pressure difference complies with the CFD reference pressure difference, which yields an R -square of 0.985 and an RMSE of 3.03 Pa. The precision of the method is tested on experimental data, measured on a carotid phantom of the common carotid artery. The volume profile used during measurement was set to mimic flow in the carotid artery with a peak flow rate of 12.9 mL/s. The experimental setup showed that the measured pressure difference changes from -59.4 to 31 Pa throughout a single pulse cycle. This was estimated with a precision of 5.44% (3.22 Pa) across ten pulse cycles. The method was also compared to invasive catheter measurements in a phantom with a 60% cross-sectional area reduction. The ultrasound method detected a maximum pressure difference of 72.3 Pa with a precision of 3.3% (2.22 Pa). The catheters measured a maximum pressure difference of 105 Pa with a precision of 11.2% (11.4 Pa). This was measured over the same constriction and with a peak flow rate of 12.9 mL/s. The double cross-correlation approach revealed no improvement compared to a normal differential operator. The method's strength, thus, lies primarily in the ultrasound sequence, which allows precise and accurate velocity estimations, at which acceleration and pressure differences can be acquired.