<Emphasis Type="Italic">In Vivo</Emphasis> MR Imaging of Dual MRI Reporter Genes and Deltex-1 Gene-modified Human Mesenchymal Stem Cells in the Treatment of Closed Penile Fracture

Purpose

The purpose of this study was to investigate the feasibility of dual magnetic resonance imaging (MRI) reporter genes, including ferritin heavy subunit (Fth) and transferrin receptor (TfR), which provide sufficient MRI contrast for in vivo MRI tracking, and the Deltex-1 (DTX1) gene, which promotes human mesenchymal stem cell (hMSC) differentiation to smooth muscle cells (SMCs), to treat closed penile fracture (CPF).

Methods

Multi-gene co-expressing hMSCs were generated. The expression of mRNA and proteins was assessed, and the original biological properties of hMSCs were determined and compared. The intracellular uptake of iron was evaluated, and the ability to differentiate into SMCs was detected. Fifty rabbits with CPF were randomly transplanted with PBS, hMSCs, Fth-TfR-hMSCs, DTX1-hMSCs, and Fth-TfR-DTX1-hMSCs. In vivo MRI was performed to detect the distribution and migration of the grafted cells and healing progress of CPF, and the results were correlated with histology.

Results

The mRNA and proteins of the multi-gene were highly expressed. The transgenes could not influence the original biological properties of hMSCs. The dual MRI reporter genes increased the iron accumulation capacity, and the DTX1 gene promoted hMSC differentiation into SMCs. The distribution and migration of the dual MRI reporter gene-modified hMSCs, and the healing state of CPF could be obviously detected by MRI and confirmed by histology.

Conclusion

The dual MRI reporter genes could provide sufficient MRI contrast, and the distribution and migration of MSCs could be detected in vivo. The DTX1 gene can promote MSC differentiation into SMCs for the treatment of CPF and effectively inhibit granulation tissue formation.

     

Multimodal Functional Neuroimaging by Simultaneous BOLD fMRI and Fiber-Optic Calcium Recordings and Optogenetic Control

Recent developments of optogenetic tools and fluorescence-based calcium recording techniques enable the manipulation and monitoring of neural circuits on a cellular level. Non-invasive imaging of brain networks, however, requires the application of methods such as blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI), which is commonly used for functional neuroimaging. While BOLD fMRI provides brain-wide non-invasive reading of the hemodynamic response, it is only an indirect measure of neural activity. Direct observation of neural responses requires electrophysiological or optical methods. The latter can be combined with optogenetic control of neuronal circuits and are MRI compatible. Yet, simultaneous optical recordings are still limited to fiber-optic-based approaches. Here, we review the integration of optical recordings and optogenetic manipulation into fMRI experiments. As a practical example, we describe how BOLD fMRI in a 9.4-T small animal MR scanner can be combined with in vivo fiber-optic calcium recordings and optogenetic control in a multimodal setup. We present simultaneous BOLD fMRI and calcium recordings under optogenetic control in rat. We outline details about MR coil configuration, choice, and usage of opsins and chemically and genetically encoded calcium sensors, fiber implantation, appropriate light power for stimulation, and calcium signal detection, to provide a glimpse into challenges and opportunities of this multimodal molecular neuroimaging approach.     

Study of Vesicular Monoamine Transporter 2 in Myopic Retina Using [<Superscript>18</Superscript>F]FP-(+)-DTBZ

Purpose

To investigate the relationship between expression level of vesicular monoamine transporter 2 (VMAT2) and myopia, as well as the feasibility of noninvasive myopia diagnosis through imaging VMAT2 in retina by using [¹⁸F]fluoropropyl-(+)-dihydrotetrabenazine ([¹⁸F]FP-(+)-DTBZ).

Procedures

The right eyes of ten guinea pigs were deprived of vision to establish form-deprived (FD) myopia and the left eyes were untreated as the self-control eyes. The location and expression level of VMAT2 in the eyes were detected by micro-positron emission tomography (PET)/X-ray computed tomography (CT) imaging through using [¹⁸F]FP-(+)-DTBZ. Immunofluorescence staining and Western blot were used to confirm the location and expression level of VMAT2 in the eyes. The concentrations of dopamine (DA) and its metabolites including 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were also investigated by high-performance liquid chromatography.

Results

The right eyes deprived of vision were obviously myopic (??3.17?±?1.33 D) after procedure, while the left eyes were hyperopic (4.60?±?0.83 D, P?<?0.0001). The main expressions of VMAT2 in the eyes were located in retina. VMAT2 was significantly reduced in the myopic retina compared to the normal one from PET/CT results (P?=?0.0008), which could also be verified by Western blots (P?=?0.029). The concentrations of DA, DOPAC, and HVA in the FD eyes were all significantly less than those in the control eyes (P?=?0.024, P?=?0.018, P?=?0.008). As a role of storing and releasing DA in vesicles, VMAT2 was demonstrated positively correlating with the amounts of DA (P?=?0.030), DOPAC (P?=?0.038), and HVA (P?=?0.025) through Pearson’s correlation coefficient test.

Conclusions

We demonstrate that [¹⁸F]FP-(+)-DTBZ can be used to noninvasively image VMAT2 in retina. The expression level of VMAT2 in retina may act as a new biomarker for myopia diagnosis. The decreasing of VMAT2 expression level may play an important role in the development of myopia through correspondingly reducing the amount of DA in retina.

     

Multispectral Photoacoustic Imaging of Tumor Protease Activity with a Gold Nanocage-Based Activatable Probe

Purpose

Tumor proteases have been recognized as significant regulators in the tumor microenvironment, but the current strategies for in vivo protease imaging have tended to focus on the development of a probe design rather than the investigation of a novel imaging strategy by leveraging the imaging technique and probe. Herein, it is the first report to investigate the ability of multispectral photoacoustic imaging (PAI) to estimate the distribution of protease cleavage sites inside living tumor tissue by using an activatable photoacoustic (PA) probe.

Procedures

The protease MMP-2 is selected as the target. In this probe, gold nanocages (GNCs) with an absorption peak at ~?800 nm and fluorescent dye molecules with an absorption peak at ~?680 nm are conjugated via a specific enzymatic peptide substrate. Upon enzymatic activation by MMP-2, the peptide substrate is cleaved and the chromophores are released. Due to the different retention speeds of large GNCs and small dye molecules, the probe alters its intrinsic absorption profile and produces a distinct change in the PA signal. A multispectral PAI technique that can distinguish different chromophores based on intrinsic PA spectral signatures is applied to estimate the signal composition changes and indicate the cleavage interaction sites. Finally, the multispectral PAI technique with the activatable probe is tested in solution, cultured cells, and a subcutaneous tumor model in vivo.

Results

Our experiment in solution with enzyme ± inhibitor, cell culture ± inhibitor, and in vivo tumor model with administration of the developed probe ± inhibitor demonstrated the probe was cleaved by the targeted enzyme. Particularly, the in vivo estimation of the cleavage site distribution was validated with the result of ex vivo immunohistochemistry analysis.

Conclusions

This novel synergy of the multispectral PAI technique and the activatable probe is a potential strategy for the distribution estimation of tumor protease activity in vivo.

     

Intravascular ultrasound assessment of the effect of laser energy on the arterial wall during the treatment of femoro-popliteal lesions: a CliRpath excimer laser system to enlarge lumen openings (CELLO) registry study

The CliRpath Excimer Laser System to Enlarge Lumen Openings (CELLO) registry included patients treated with modified excimer laser catheters for the endovascular treatment of peripheral artery disease affecting the superficial femoral artery (SFA) and proximal popliteal artery. The aim of this study was to assess, via intravascular ultrasound (IVUS) the dissections in the vessel wall following treatment with the laser catheters. IVUS grayscale images from the CELLO registry were systematically reviewed for dissections in the treated vessel segments by two investigators. Images from 33 patients; 66 pullbacks (1867 IVUS frames in 2 phases), were successfully matched frame-to-frame to evaluate identical segments of the treated vessels in the two phases; post-2 mm Turbo-Elite laser pilot channel creation and post Turbo-Booster laser atherectomy. Dissections were categorized as; (1) intimal, (2) medial, (3) intramural hematoma, and (4) adventitial according to the ACC Clinical Expert Consensus Document classification of dissections. An average of 57 frames was evaluated per pullback, giving a total of 3734 frames (1867 matched for pre-ablation (post channel creation) and post-ablation phases). Treatments with the modified Excimer laser catheters resulted in a significant increase in lumen area of 5.5?±?3.2-mm² (95% CI 4.3–6.8, p?<?0.0001) and reduction in plaque plus media volume of ?10.6?±?36.0 mm³ (95% CI ?25.8 to 4.6, p?=?0.1619) whilst giving rise to mainly intramural hematoma formations post Turbo-Booster laser treatment in 55% of frames assessed and 24% medial dissections with less than 1% adventitial disruption. The Excimer laser based Turbo-Booster treatment of peripheral artery lesions resulted in significant plaque debulking and increased lumen diameter with negligible degree of adventitial layer injury.     

Use of speckle tracking in the evaluation of late subclinical myocardial damage in survivors of childhood acute leukaemia

Heart disease is the leading cause of non-cancer death in childhood cancer survivors. to determine the prevalence of subclinical cardiac dysfunction using speckle tracking and compare its results with those obtained by classical methods of assessing left ventricular function and its relationship with different factors to identify the population at higher risk. Echocardiographic assessment of left ventricular function included ejection fraction, tissue Doppler, longitudinal/circumferential strains and biochemical parameters (troponin-T and Pro-BNP) in a cohort of 57 survivors of childhood acute leukaemia with at least 10 years since diagnosis. Ventricular dysfunction was found in 5.2% of patients in M-mode (ejection fraction—EF?<?53% with a reduction in the EF?≥?10%) and in 7% of patients with Simpson’s method, compared with 21.05 and 8.8% with suboptimal global longitudinal strain (GLS) and global circumferential strain, respectively. The GLS alteration was significantly correlated with lower values of left ventricular systolic function and was associated with high tumour risk (odds ratio [OR] 13.8), cumulative doses of anthracyclines?≥?250 mg/m² (OR 7.6) and radiotherapy (OR 7.19). Biomarkers were not useful for the diagnosis of subclinical cardiomyopathy. Good reproducibility was obtained, with an intraobserver correlation of 93.6% and an interobserver correlation of 89.2% in the GLS. The alteration of the GLS was more prevalent than the alteration in the EF and was associated with the treatment received and high tumour risk. strain imaging seems to be a powerful tool to identify an increased number of survivor with an early myocardial injury.     

Incidental findings in multislice computed tomography prior to transcatheter aortic valve implantation: frequency,clinical relevance and outcome

Multislice computed tomography (MSCT) has emerged as the mainstay in patients planned for transcatheter aortic valve implantation (TAVI). Incidental findings (IF) in MSCT are common. However, the exact incidence, clinical relevance and further consequences of IF are unclear and it is controversial whether IF adversely affect patients’ outcome. We analyzed MSCT data of 1050 patients screened for TAVI between January 2011 and December 2014. Median follow-up of patients was 20 months. In total, 3194 IF were identified, which were classified into clinically non-relevant IF (2872, 90%) and clinically relevant IF (322, 10%). In 25% of patients (258/1050) at least one clinically relevant IF was present. Age (80?±?7 vs. 80?±?7 years; p?=?0.198) and EuroSCORE II (3.6% [2.1–5.7] vs. 3.6% [2.1–5.9]; p?=?0.874) was similar between patients with and without a clinically relevant IF. TAVI was performed less frequently in patients with a clinically relevant IF (76% vs. 85%; p?<?0.001), with more patients receiving surgical aortic valve replacement in that group (14% vs. 11%; p?=?0.042), possibly due to the high rate of incidental aneurysms of the ascending aorta (n?=?48). If TAVI was performed mortality did not differ (30-days: 4% vs. 3%; p?=?0.339, 1-year: 11% vs. 14%; p?=?0.226) between patients with and without a clinically relevant IF. Our study is the largest study to analyze prevalence, clinical relevance and therapeutic consequences of IF during screening for TAVI. IF in pre-procedural MSCT are common and clinically relevant in one-quarter of patients. However, these findings had no impact on overall mortality.     

Cardiac function in patients with polymyositis or dermatomyositis: a three-dimensional speckle-tracking echocardiography study

Cardiac event is a major cause of death in patients with idiopathic inflammatory myopathies (IIM). The most frequent IIMs are polymyositis (PM) and dermatomyositis (DM). The purpose of this study was to analyze cardiac involvement by three-dimensional speckle-tracking echocardiography (3D STE) in patients with PM or DM, and to identify the relationship of cardiac injury with clinical characteristics and disease-specific parameters. 60 PM/DM patients with preserved left ventricular ejection fraction and 30 matched healthy controls were assessed by conventional echocardiography, 3D STE with biventricular strain analysis and electrocardiogram. Compared to controls, patients with PM/DM had significantly diminished left ventricular global longitudinal systolic strain and right ventricular longitudinal systolic strain (LVGLS, ? 20.3?±?2.5 vs. ? 23.4?±?1.7%; RVLS, ? 19.4?±?4.2 vs ? 24.8?±?2.0%; both P?<?0.001), and longer QTc intervals(421.0?±?38.4 vs 400.6?±?14.5 ms, P?=?0.001). Multiple regression analysis showed that Myositis Damage Index (MDI) was independently associated with LVGLS (R²?=?0.44, P?=?0.002) and RVLS (R²?=?0.56, P?<?0.001) in PM/DM patients with established disease course more than 1 year. In multivariate analysis of pooled data for all the PM/DM patients, when MDI was excluded due to missing observations, disease duration correlated with worse LVGLS (R²?=?0.24, P?=?0.002), while concomitant interstitial lung disease correlated with worse RVLS (R²?=?0.30, P?<?0.001). Disease activity scores (Myositis Intention to Treat Activities Index) had a weak positive correlation with QTc intervals (r_sp = 0.31, P?=?0.02). Our results suggest that cardiac injury in PM/DM is a long-term process and its severity depends on patients’ heterogeneous clinical features and systemic disease burden.     

Association with right atrial strain with right atrial pressure: an invasive validation study

Echocardiographic assessment of right atrial pressure (RAP) from inferior vena cava (RAP_IVC) dimension may underestimate catheter-derived (RAP_C). As right atrial (RA) deformation, measured by speckle tracking, is preload-dependent, we hypothesized that RA strain may improve estimation of RAP_C. Right atrial strain components [RA reservoir function (?R), peak RA contraction (?CT) and RA conduit function (?CD)] were measured in 125 of 175 patients who had echocardiography and invasive measures of RAP (median difference 1 day). To determine whether RA strain measures differentiated patients with correct vs incorrect RAP_IVC assessment, categories with RAP_IVC values?<?3, 8 and >?15 mmHg were compared with RAP_C groups?<?3, 4–7, 8–10, 11–14 and >?15 mmHg. Non-invasively determined RAP was significantly lower (p?=?0.001) than invasively determined RAP_C, with a weak correlation (r?=?0.35, p?<?0.001). RA strain components were associated with RA size, RV function and IVC size. In those with RAP_IVC?>?15 mmHg, half of patients were categorized into RAP?<?10 mmHg. There were no significant differences in RA characteristics that differentiated patients in whom echocardiographic estimation of RAP was inaccurate. Right atrial strain measures were feasible, and had associations with RA size, RV systolic function and IVC size. Right atrial strain was significantly different between those with normal vs raised pressure, but it did not identify those with incorrect echocardiographic assessment of RAP.     

Variability of native T1 values: implication for defining regional myocardial changes using MRI

The aim of the present study was to establish T1 variation (T1v) thresholds for duplicated measurements of regional T1 values in left ventricle (LV) using magnetic resonance imaging (MRI). Eighteen healthy volunteers were recruited to undergo two consecutive cardiac MRI scans using modified Look-Locker Inversion recovery (MOLLI) with two spatial resolutions on different days to repeat T1 measurements on LV. The absolute differences (d) and standard deviations (SDs) of regional T1 values were acquired with the two scans and two readers. T1v threshold (mean difference?+?2SD), intra-class correlation coefficient (ICC) and coefficient of variation (CoV) were calculated. T1 mapping using the MOLLI sequence (with multiple spatial resolutions) was successfully performed in all 18 volunteers twice. On a per-slice basis, ICCs for intra-observer, inter-observer, inter-resolution and inter-study T1v were 0.988, 0.899, 0.763 and 0.6. CoVs were 0.72, 2.39, 3.90 and 4.28%. T1v thresholds were 22, 66, 118 and 120 ms. On a per-segment basis, ICCs for intra-observer, inter-observer, inter-resolution and inter-study T1v were 0.974, 0.859, 0.711 and 0.594. CoVs were 1.09, 3.36, 4.69 and 5.01%. T1v thresholds were 33, 94, 140 and 144 ms. Those thresholds may be useful for discriminating disease-initiated T1v from random errors of T1 measurements.