Fetal dynamic phase-contrast MR angiography using ultrasound gating and comparison with Doppler ultrasound measurements.

OBJECTIVES: To investigate the feasibility of fetal phase-contrast (PC)-MR angiography of the descending aorta (AoD) using an MR-compatible Doppler ultrasound sensor (DUS) for fetal cardiac gating and to compare velocimetry with Doppler ultrasound measurements. METHODS: In this prospective study, 2D PC-MR angiography was performed in 12 human fetuses (mean gestational age 32.8 weeks) using an MR-compatible DUS for gating of the fetal heart at 1.5 T. Peak flow velocities in the fetal AoD were compared with Doppler ultrasound measurements performed on the same day. Reproducibility of PC-MR measurements was tested by repeated PC-MR in five fetuses. RESULTS: Dynamic PC-MR angiography in the AoD was successfully performed in all fetuses using the DUS, with an average fetal heart rate of 140 bpm (range 129-163). Time-velocity curves revealed typical arterial blood flow patterns. PC-MR mean flow velocity and mean flux were 21.2 cm/s (range 8.6-36.8) and 8.4 ml/s (range 3.2-14.6), respectively. A positive association between PC-MR mean flux and stroke volume with gestational age was obtained (r = 0.66, p = 0.02 and r = 0.63, p = 0.03). PC-MR and Doppler ultrasound peak velocities revealed a highly significant correlation (r = 0.8, p < 0.002). Peak velocities were lower for PC-MR with 69.1 cm/s (range 39-125) compared with 96.7 cm/s (range 60-142) for Doppler ultrasound (p < 0.001). Reproducibility of PC-MR was high (p > 0.05). CONCLUSION: The MR-compatible DUS for fetal cardiac gating allows for PC-MR angiography in the fetal AoD. Comparison with Doppler ultrasound revealed a highly significant correlation of peak velocities with underestimation of PC-MR velocities. This new technique for direct fetal cardiac gating indicates the potential of PC-MR angiography for assessing fetal hemodynamics. KEY POINTS: • The developed MR-compatible Doppler ultrasound sensor allows direct fetal cardiac gating and can be used for prenatal dynamic cardiovascular MRI. • The MR-compatible Doppler ultrasound sensor was successfully applied to perform intrauterine phase-contrast MR angiography of the fetal aorta, which revealed a highly significant correlation with Doppler ultrasound measurements. • As fetal flow hemodynamics is an important parameter in the diagnosis and management of fetal pathologies, fetal phase-contrast MR angiography may offer an alternative imaging method in addition to Doppler ultrasound and develop as a second line tool in the evaluation of fetal flow hemodynamics.

Dynamic fetal cardiac magnetic resonance imaging in four-chamber view using Doppler ultrasound gating in normal fetal heart and in congenital heart disease: comparison with fetal echocardiography.

OBJECTIVES: To investigate the diagnostic performance of dynamic fetal cardiac magnetic resonance imaging (MRI), using a MR-compatible Doppler ultrasound (DUS) device for fetal cardiac gating, in differentiating fetuses with congenital heart disease from those with a normal heart, and to compare the technique with fetal echocardiography. METHODS: This was a prospective study of eight fetuses with a normal heart and four with congenital heart disease (CHD), at a median of 34 (range, 28-36) weeks' gestation. Dynamic fetal cardiac MRI was performed using a DUS device for direct cardiac gating. The four-chamber view was evaluated according to qualitative findings. Measurements of the length of the left and right ventricles, diameter of the tricuspid and mitral valves, myocardial wall thickness, transverse cardiac diameter and left ventricular planimetry were performed. Fetal echocardiography and postnatal diagnoses were considered the reference standards. RESULTS: Direct cardiac gating allowed continuous triggering of the fetal heart, showing high temporal and spatial resolution. Both fetal cardiac MRI and echocardiography in the four-chamber view detected pathological findings in three of the 12 fetuses. Qualitative evaluation revealed overall consistency between echocardiography and MRI. On both echocardiography and MRI, quantitative measurements revealed significant differences between fetuses with a normal heart and those with CHD with respect to the length of the right (P < 0.01 for both) and left (P < 0.01 for both) ventricles and transverse cardiac diameter (P < 0.05 and P < 0.01, respectively). Tricuspid valve diameter on cardiac MRI was found to be significantly different in healthy fetuses from in those with CHD (P < 0.05). CONCLUSIONS: For the first time, this study has shown that dynamic fetal cardiac MRI in the four-chamber view, using external cardiac gating, allows evaluation of cardiac anatomy and diagnosis of congenital heart disease in agreement with fetal echocardiography. Dynamic fetal cardiac MRI may be useful as a second-line investigation if conditions for fetal echocardiography are unfavorable. Copyright © 2018 ISUOG. Published by John Wiley & Sons Ltd.

Doppler ultrasound triggering for cardiac MRI at 7T.

PURPOSE: A prerequisite for cardiac MR (CMR) imaging is adequate synchronization of image acquisition with the cardiac cycle. Electrocardiogram triggering may be hampered by electromagnetic interferences at high field strength. The purpose of this work is to evaluate the feasibility of Doppler ultrasound triggering for CMR image synchronization at 7T ultra-high-field MRI. METHODS: A custom-built Doppler ultrasound (DUS) trigger device was developed. Magnetic resonance compatibility was evaluated using E- and H-field probes and flip angle maps prior to the study. Cardiac MR was performed at 7T in 13 healthy subjects using DUS and pulse oximetry for triggering. For validation of the trigger signal, the electrocardiogram, pulse, and DUS signals were compared outside of the MR room. Breath-hold cine fast low-angle-shot sequences were acquired in short-axis and four-chamber view. Image quality was assessed by two senior radiologists and by measurement of endocardial blurring. RESULTS: The maximal change in E- and H-field distributions with and without transducer was 5%. No interferences were observed between DUS and MRI in the B1 maps and during CMR imaging. Validation of the DUS trigger signal resulted in a high correlation to the electrocardiographic signal of r = 0.99. Analysis of image and trigger quality revealed no significant differences. CONCLUSION: Doppler ultrasound was applied as a new trigger method in CMR at 7T. The transmission line and transducer were locally approved as 7T MR conditional, and were successfully tested for image synchronization at 7T. In the future, this method needs to be evaluated in a larger patient population. Magn Reson Med 80:239-247, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

One-step, highly efficient site-directed mutagenesis by toxic protein selection.

A fast and efficient site-directed mutagenesis method has been developed, using the newly constructed plasmid pTPS19, which expresses the toxic CcdB protein originally encoded by the E. coli F plasmid. Once the target gene is cloned into pTPS19, desired mutations can be introduced with two primers. The first contains the desired mutation, and the second is designed to create a +1 frame shift in the ccdB gene to inactivate the CcdB protein. The mutants can be directly selected on LB plates containing IPTG, through which the toxic CcdB protein is induced, thereby eliminating cells carrying wild-type parental plasmids. Based on stringent selection through the toxic CcdB protein, mutagenesis efficiency of 90%-100% was reached even after one round of transformation.

Molecularly cloned SHIV-1157ipd3N4: a highly replication- competent, mucosally transmissible R5 simian-human immunodeficiency virus encoding HIV clade C Env.

Human immunodeficiency virus type 1 (HIV-1) clade C causes >50% of all HIV infections worldwide, and an estimated 90% of all transmissions occur mucosally with R5 strains. A pathogenic R5 simian-human immunodeficiency virus (SHIV) encoding HIV clade C env is highly desirable to evaluate candidate AIDS vaccines in nonhuman primates. To this end, we generated SHIV-1157i, a molecular clone from a Zambian infant isolate that carries HIV clade C env. SHIV-1157i was adapted by serial passage in five monkeys, three of which developed peripheral CD4(+) T-cell depletion. After the first inoculated monkey developed AIDS at week 137 postinoculation, transfer of its infected blood to a naïve animal induced memory T-cell depletion and thrombocytopenia within 3 months in the recipient. In parallel, genomic DNA from the blood donor was amplified to generate the late proviral clone SHIV-1157ipd3. To increase the replicative capacity of SHIV-1157ipd3, an extra NF-kappaB binding site was engineered into its 3' long terminal repeat, giving rise to SHIV-1157ipd3N4. This virus was exclusively R5 tropic and replicated more potently in rhesus peripheral blood mononuclear cells than SHIV-1157ipd3 in the presence of tumor necrosis factor alpha. Rhesus macaques of Indian and Chinese origin were next inoculated intrarectally with SHIV-1157ipd3N4; this virus replicated vigorously in both sets of monkeys. We conclude that SHIV-1157ipd3N4 is a highly replication-competent, mucosally transmissible R5 SHIV that represents a valuable tool to test candidate AIDS vaccines targeting HIV-1 clade C Env.