L-Wnk1 Deletion in Smooth Muscle Cells Causes Aortitis and Inflammatory Shift.

BACKGROUND: The long isoform of the Wnk1 (with-no-lysine [K] kinase 1) is a ubiquitous serine/threonine kinase, but its role in vascular smooth muscle cells (VSMCs) pathophysiology remains unknown. METHODS: AngII (angiotensin II) was infused in Apoe-/- to induce experimental aortic aneurysm. Mice carrying an Sm22-Cre allele were cross-bred with mice carrying a floxed Wnk1 allele to specifically investigate the functional role of Wnk1 in VSMCs. RESULTS: Single-cell RNA-sequencing of the aneurysmal abdominal aorta from AngII-infused Apoe-/- mice revealed that VSMCs that did not express Wnk1 showed lower expression of contractile phenotype markers and increased inflammatory activity. Interestingly, WNK1 gene expression in VSMCs was decreased in human abdominal aortic aneurysm. Wnk1-deficient VSMCs lost their contractile function and exhibited a proinflammatory phenotype, characterized by the production of matrix metalloproteases, as well as cytokines and chemokines, which contributed to local accumulation of inflammatory macrophages, Ly6Chi monocytes, and γδ T cells. Sm22Cre+Wnk1lox/lox mice spontaneously developed aortitis in the infrarenal abdominal aorta, which extended to the thoracic area over time without any negative effect on long-term survival. AngII infusion in Sm22Cre+Wnk1lox/lox mice aggravated the aortic disease, with the formation of lethal abdominal aortic aneurysms. Pharmacological blockade of γδ T-cell recruitment using neutralizing anti-CXCL9 antibody treatment, or of monocyte/macrophage using Ki20227, a selective inhibitor of CSF1 receptor, attenuated aortitis. Wnk1 deletion in VSMCs led to aortic wall remodeling with destruction of elastin layers, increased collagen content, and enhanced local TGF-ß (transforming growth factor-beta) 1 expression. Finally, in vivo TGF-ß blockade using neutralizing anti-TGF-ß antibody promoted saccular aneurysm formation and aorta rupture in Sm22 Cre+ Wnk1lox/lox mice but not in control animals. CONCLUSION: Wnk1 is a key regulator of VSMC function. Wnk1 deletion promotes VSMC phenotype switch toward a pathogenic proinflammatory phenotype, orchestrating deleterious vascular remodeling and spontaneous severe aortitis in mice.

FIBER-ML, an Open-Source Supervised Machine Learning Tool for Quantification of Fibrosis in Tissue Sections.

Pathologic fibrosis is a major hallmark of tissue insult in many chronic diseases. Although the amount of fibrosis is recognized as a direct indicator of the extent of disease, there is no consentaneous method for its quantification in tissue sections. This study tested FIBER-ML, a semi-automated, open-source freeware that uses a machine-learning approach to quantify fibrosis automatically after a short user-controlled learning phase. Fibrosis was quantified in sirius red-stained tissue sections from two fibrogenic animal models: acute stress-induced cardiomyopathy in rats (Takotsubo syndrome-like) and HIV-induced nephropathy in mice (chronic kidney disease). The quantitative results of FIBER-ML software version 1.0 were compared with those of ImageJ in Takotsubo syndrome, and with those of inForm in chronic kidney disease. Intra- and inter-operator and inter-software correlation and agreement were assessed. All correlations were excellent (>0.95) in both data sets. The values of discriminatory power between the pathologic and healthy groups were <10-3 for data on Takotsubo syndrome and <10-4 for data on chronic kidney disease. Intra-operator agreement, assessed by intra-class coefficient correlation, was good (>0.8), while inter-operator and inter-software agreement ranged from moderate to good (>0.7). FIBER-ML performed in a fast and user-friendly manner, with reproducible and consistent quantification of fibrosis in tissue sections. It offers an open-source alternative to currently used software, including quality control and file management.

The BMS-LM ontology for biomedical data reporting throughout the lifecycle of a research study: From data model to ontology.

Biomedical research data reuse and sharing is essential for fostering research progress. To this aim, data producers need to master data management and reporting through standard and rich metadata, as encouraged by open data initiatives such as the FAIR (Findable, Accessible, Interoperable, Reusable) guidelines. This helps data re-users to understand and reuse the shared data with confidence. Therefore, dedicated frameworks are required. The provenance reporting throughout a biomedical study lifecycle has been proposed as a way to increase confidence in data while reusing it. The Biomedical Study - Lifecycle Management (BMS-LM) data model has implemented provenance and lifecycle traceability for several multimodal-imaging techniques but this is not enough for data understanding while reusing it. Actually, in the large scope of biomedical research, a multitude of metadata sources, also called Knowledge Organization Systems (KOSs), are available for data annotation. In addition, data producers uses local terminologies or KOSs, containing vernacular terms for data reporting. The result is a set of heterogeneous KOSs (local and published) with different formats and levels of granularity. To manage the inherent heterogeneity, semantic interoperability is encouraged by the Research Data Management (RDM) community. Ontologies, and more specifically top ontologies such as BFO and DOLCE, make explicit the metadata semantics and enhance semantic interoperability. Based on the BMS-LM data model and the BFO top ontology, the BioMedical Study - Lifecycle Management (BMS-LM) core ontology is proposed together with an associated framework for semantic interoperability between heterogeneous KOSs. It is made of four ontological levels: top/core/domain/local and aims to build bridges between local and published KOSs. In this paper, the conversion of the BMS-LM data model to a core ontology is detailed. The implementation of its semantic interoperability in a specific domain context is explained and illustrated with examples from small animal preclinical research.

Radiomic analysis of HTR-DCE MR sequences improves diagnostic performance compared to BI-RADS analysis of breast MR lesions.

PURPOSE: To assess the diagnostic performance of radiomic analysis using high temporal resolution (HTR)-dynamic contrast enhancement (DCE) MR sequences compared to BI-RADS analysis to distinguish benign from malignant breast lesions. MATERIALS AND METHODS: We retrospectively analyzed data from consecutive women who underwent breast MRI including HTR-DCE MR sequencing for abnormal enhancing lesions and who had subsequent pathological analysis at our tertiary center. Semi-quantitative enhancement parameters and textural features were extracted. Temporal change across each phase of textural features in HTR-DCE MR sequences was calculated and called "kinetic textural parameters." Statistical analysis by LASSO logistic regression and cross validation was performed to build a model. The diagnostic performance of the radiomic model was compared to the results of BI-RADS MR score analysis. RESULTS: We included 117 women with a mean age of 54 years (28-88). Of the 174 lesions analyzed, 75 were benign and 99 malignant. Seven semi-quantitative enhancement parameters and 57 textural features were extracted. Regression analysis selected 15 significant variables in a radiomic model (called "malignant probability score") which displayed an AUC = 0.876 (sensitivity = 0.98, specificity = 0.52, accuracy = 0.78). The performance of the malignant probability score to distinguish benign from malignant breast lesions (AUC = 0.876, 95%CI 0.825-0.925) was significantly better than that of BI-RADS analysis (AUC = 0.831, 95%CI 0.769-0.892). The radiomic model significantly reduced false positives (42%) with the same number of missed cancers (n = 2). CONCLUSION: A radiomic model including kinetic textural features extracted from an HTR-DCE MR sequence improves diagnostic performance over BI-RADS analysis. KEY POINTS: • Radiomic analysis using HTR-DCE is of better diagnostic performance (AUC = 0.876) than conventional breast MRI reading with BI-RADS (AUC = 0.831) (p < 0.001). • A radiomic malignant probability score under 19.5% gives a negative predictive value of 100% while a malignant probability score over 81% gives a positive predictive value of 100%. • Kinetic textural features extracted from HTR-DCE-MRI have a major role to play in distinguishing benign from malignant breast lesions.

Assessment of BOLD response in the fetal lung.

OBJECTIVE: Assessment of lung development and maturity is of utmost importance in prenatal counseling. Blood oxygen level-dependent (BOLD) effect MRI was developed for functional evaluations of organs. To date, no data are available in fetal lungs and nothing is known about the existence of a BOLD effect in the lungs. The aim of our study was to evaluate if a BOLD response could be detected in fetal lungs. MATERIALS AND METHODS: From January 2014 to December 2016, 38 healthy pregnant women were prospectively enrolled. After a routine scan on a 1.5-T MRI device (normoxic period), maternal hyperoxia was induced for 5 min before the BOLD sequence (hyperoxic period). R2* was evaluated by fitting average intensity of the signal, both for normoxic (norm) and hyperoxic (hyper) periods. RESULTS: A significant BOLD response was observed after maternal hyperoxia in the lungs with a mean R2* decrease of 12.1 ± 2.5% (p < 0.001), in line with the placenta response with a mean R2* decrease of 19.2 ± 5.9% (p < 0.0001), confirming appropriate oxygen uptake. Conversely, no significant BOLD effect was observed for the brain nor the liver with a mean ∆R2* of 3.6 ± 3.1% (p = 0.64) and 2.8 ± 3.7% (p = 0.23). CONCLUSION: This study shows for the first time in human that a BOLD response can be observed in the normal fetal lung despite its prenatal "non-functional status." If confirmed in congenital lung and chest malformations, this property could be used in addition to the lung volume for a better prediction of postnatal respiratory status. KEY POINTS: • Blood oxygen level-dependent (BOLD) effect MRI was developed for functional evaluations of organs and could have interesting implications for the fetal organs. • Assessment of lung development is of utmost importance in prenatal counseling, but to date no data are available in fetal lungs. • BOLD response can be observed in the normal fetal lung opening the way to studies on fetus with pathological lungs.

Quality-based pharmacokinetic model selection on DCE-MRI for characterizing orbital lesions.

BACKGROUND: Although several studies have evaluated dynamic contrast-enhanced (DCE) MRI in the orbit, showing its utility when detecting and diagnosing orbital lesions, none have evaluated the pharmacokinetic models. PURPOSE: To provide a quality-based pharmacokinetic model selection for characterizing orbital lesions using DCE-MRI at 3.0T. STUDY TYPE: Prospective. POPULATION: From December 2015 to April 2017, 151 patients with an orbital lesion underwent MRI prior to surgery, including a high temporal resolution DCE sequence, divided into one training and one test dataset with 100 and 51 patients, respectively. FIELD STRENGTH/SEQUENCE: 3T/DCE. ASSESSMENT: Six different pharmacokinetic models were tested. STATISTICAL TESTS: Univariate and multivariate analyses were performed using Wilcoxon-2-sample tests and a logistic regression to compare parameters between malignant and benign tumors for each pharmacokinetic model for the whole cohort. Receiver operating characteristic (ROC) curve analyses were performed on the training dataset to determine area under curve (AUC) and optimal cutoff values for each pharmacokinetic model, then validated on the test dataset to calculate sensitivity, specificity, and accuracy. RESULTS: Regardless of the model, tissue blood flow and tissue blood volume values were significantly higher in malignant vs. benign lesions: 103.8-195.1 vs. 65-113.8, P [<10-4 -2.10-4 ] and 21.3-36.9 vs. 15.6-33.6, P [<10-4 -0.03] respectively. Extracellular volume fraction and permeability-surface area product or transfer constant appeared to be less relevant: 17.3-27.5 vs. 22.8-28.2, P [0.01-0.7], 1.7-4.9, P [0.2-0.9] and 9.5-38.8 vs. 8.1-22.8, P [<10-4 -0.6], respectively. ROC curves showed no significant differences in AUC between the different models. The two-compartment exchange (2CX) model ranked first for quality. DATA CONCLUSION: DCE MRI pharmacokinetic model-derived parameters appeared to be useful for discriminating benign from malignant orbital lesions. The 2CX model provided the best quality of modeling and should be recommended. Perfusion-related DCE parameters appeared to be significantly more relevant to the diagnostic process. Level of Evidence 1 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;50:1514-1525.

Assessment of Placental Perfusion in the Preeclampsia L-NAME Rat Model with High-Field Dynamic Contrast-Enhanced MRI.

PURPOSE: To evaluate placental function and perfusion in a rat model of preeclampsia infused with L-nitro-arginine methyl ester (L-NAME) by dynamic contrast-enhanced (DCE) MRI using gadolinium chelates. METHODS: Pregnant female Sprague-Dawley rats were fitted on embryonic day 16 (E16) with subcutaneous osmotic minipumps loaded to deliver, continuously, L-NAME (50 mg/day per rat; case group) or saline solution (control group). DCE MRI was performed on E19 using gadolinium chelates and a 4.7-T MRI apparatus for small animals. Quantitative analysis was performed using an image software program: placental blood flow (perfusion in mL/min/100 mL of placenta) and fractional volume of the maternal vascular placental compartment (ratio between the placental blood volume and the placental volume, Vb in %) were calculated by compartmental analysis. RESULTS: A total of 176 placentas (27 rats) were analyzed by DCE MRI (97 cases and 79 controls). The model was effective, inducing intrauterine growth retardation, as there was a significant difference between the two groups for placental weight (p < 0.01), fetal weight (p = 0.019), and fetal length (p < 0.01). There was no significant difference in placental perfusion between the L-NAME and control groups (140.1 ± 74.1 vs. 148.9 ± 97.4, respectively; p = 0.496). There was a significant difference between the L-NAME and control groups for Vb (53 ± 12.9 vs. 46.7 ± 9%, respectively; p < 0.01). CONCLUSION: In the L-NAME preeclampsia model, placental perfusion is normal and the fractional blood volume is increased, suggesting that preeclampsia is not always expressed as a result of decreased placental perfusion. This highlights the usefulness of MRI for investigating the physiopathology of preeclampsia.

Quantifying tumor vascular heterogeneity with DCE-MRI in complex adnexal masses: A preliminary study.

PURPOSE: To evaluate the value of quantifying dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) heterogeneity to characterize adnexal masses. MATERIALS AND METHODS: Our database was retrospectively queried to identify all surgically proven adnexal masses characterized with a 1.5T DCE-MRI between January 1st 2008 and February 28th 2010 (n = 113 masses, including 52 benign, 11 borderline, and 50 invasive malignant tumors). The solid component of the adnexal mass was segmented. Quantitative analysis with a compartmental model was performed to calculate microvascular parameters, including tissue blood flow (FT ), blood volume fraction (Vb ), lag time (DAT ), interstitial volume fraction (Ve ), permeability-surface area product (PS), and relative area under curve (r AUC), were calculated. Then heterogeneity parameters were evaluated using the analysis of the evolution of the standard deviation (SD) of signal intensities on DCE-MRI series. The area under the receiver operating characteristic (AUROC) curve was calculated to assess the overall discrimination of parameters. RESULTS: Malignant tumors displayed higher FT , Vb , and r AUC and lower DAT than benign tumors (P = 0.01, P < 0.0001, and P < 0.0001, respectively). Invasive malignant tumors displayed lower Vb and r AUC than borderline tumors (P < 0.01). After injection, whenever the heterogeneity parameter was considered, malignant tumors were more heterogeneous than benign tumors, invasive tumors were more heterogeneous than borderline ovarian tumors, and malignant tumors with carcinomatosis were more heterogeneous than tumors without carcinomatosis (P < 0.001). The most discriminant parameter was the SD during the 90 seconds after injection related to arterial input function (ΔSDEARLY/AIF ) with an AUROC between 0.715 and 0.808. CONCLUSION: This study proposes heterogeneity parameters as a new tool with a potential for clinical application, given that the technique uses routine imaging sequences. LEVEL OF EVIDENCE: 3 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2017;46:1776-1785.

Repeatability of apparent diffusion coefficient and intravoxel incoherent motion parameters at 3.0 Tesla in orbital lesions.

OBJECTIVES: To evaluate repeatability of intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) parameters in the orbit. METHODS: From December 2015 to March 2016, 22 patients were scanned twice using an IVIM sequence with 15b values (0-2,000 s/mm2) at 3.0T. Two readers independently delineated regions of interest in an orbital mass and in different intra-orbital and extra-orbital structures. Short-term test-retest repeatability and inter-observer agreement were assessed using the intra-class correlation coefficient (ICC), the coefficient of variation (CV) and Bland-Altman limits of agreements (BA-LA). RESULTS: Test-retest repeatability of IVIM parameters in the orbital mass was satisfactory for ADC and D (mean CV 12% and 14%, ICC 95% and 93%), poor for f and D*(means CV 43% and 110%, ICC 90% and 65%). Inter-observer repeatability agreement was almost perfect in the orbital mass for all the IVIM parameters (ICC = 95%, 93%, 94% and 90% for ADC, D, f and D*, respectively). CONCLUSIONS: IVIM appeared to be a robust tool to measure D in orbital lesions with good repeatability, but this approach showed a poor repeatability of f and D*. KEY POINTS: • IVIM technique is feasible in the orbit. • IVIM has a good-acceptable repeatability of D (CV range 12-25 %). • IVIM interobserver repeatability agreement is excellent (ICC range 90-95 %). • f or D* provide higher test-retest and interobserver variabilities.

RODES software for dose assessment of rats and mice contaminated with radionuclides.

In order to support animal experiments of chronic radionuclides intake with realistic dosimetry, voxel-based three-dimensional computer models of mice and rats of both sexes and three ages were built from magnetic resonance imaging. Radiation transport of mono-energetic photons of 11 energies and electrons of 7 energies was simulated with MCNPX 2.6c to assess specific absorbed fractions (SAFs) of energy emitted from 13 source regions and absorbed in 28 target regions. RODES software was developed to combine SAF with radiation emission spectra and user-supplied biokinetic data to calculate organ absorbed doses per nuclear transformation of radionuclides in source regions (S-factors) and for specific animal experiments with radionuclides. This article presents the design of RODES software including the simulation of the particles in the created rodent voxel phantoms. SAF and S-factor values were compared favourably with published results from similar studies. The results are discussed for rodents of different ages and sexes.

Antitumoral Effect of Mural Cells Assessed With High-Resolution MRI and Fluorescence Microscopy.

OBJECTIVE: The purpose of this study was to detect labeled mural cells in vivo and study their therapeutic effect on tumor growth and on functional changes in the vascular network by use of MRI and fibered confocal fluorescence microscopy (FCFM). MATERIALS AND METHODS: Twenty-eight mice were allocated to the following three groups 7 days after injection of TC1 tumor cells (C157 black 6): control, no injection (n = 7); sham, injection of phosphate-buffered saline solution (n = 10); and treated, injection of human mural cells (n = 11). Tumor growth was measured with calipers. Labeled mural cells were tracked with high-resolution MRI and FCFM. Microvessel density was assessed with MRI and FCFM, and the findings were compared with the histologic results. RESULTS: Tumor growth was significantly slowed in the treated group starting on day 10 (p = 0.001). Round signal-intensity voids were observed in the center of six of seven tumors treated with magnetically labeled mural cells. Positive staining for iron was observed in histologic sections of two of five of these tumors. Microvessel density measured with FCFM was greater in the treated mice (p = 0.03). Flow cytometry revealed viable human mural cells only in treated tumors. CONCLUSION: In this study, imaging techniques such as high-resolution MRI and FCFM showed the therapeutic effect of mural cell injection on tumor growth and microvessel function.

Chronic urinary obstruction: evaluation of dynamic contrast-enhanced MR urography for measurement of split renal function.

PURPOSE: To evaluate if measurement of split renal function ( SRF split renal function ) with dynamic contrast material-enhanced ( DCE dynamic contrast enhanced ) magnetic resonance (MR) urography is equivalent to that with renal scintigraphy ( RS renal scintigraphy ) in patients suspected of having chronic urinary obstruction. MATERIALS AND METHODS: The study protocol was approved by the institutional ethics committee of the coordinating center on behalf of all participating centers. Informed consent was obtained from all adult patients or both parents of children. This prospective, comparative study included 369 pediatric and adult patients from 14 university hospitals who were suspected of having chronic or intermittent urinary obstruction, and data from 295 patients with complete data were used for analysis. SRF split renal function was measured by using the area under the curve and the Patlak-Rutland methods, including successive review by a senior and an expert reviewer and measurement of intra- and interobserver agreement for each technique. An equivalence test for mean SRF split renal function was conducted with an α of 5%. RESULTS: Reproducibility was substantial to almost perfect for both methods. Equivalence of DCE dynamic contrast enhanced MR urography and RS renal scintigraphy for measurement of SRF split renal function was shown in patients with moderately dilated kidneys (P < .001 with the Patlak-Rutland method). However, in severely dilated kidneys, the mean SRF split renal function measurement was underestimated by 4% when DCE dynamic contrast enhanced MR urography was used compared with that when RS renal scintigraphy was used. Age and type of MR imaging device had no significant effect. CONCLUSION: For moderately dilated kidneys, equivalence of DCE dynamic contrast enhanced MR urography to RS renal scintigraphy was shown, with a standard deviation of approximately 12% between the techniques, making substitution of DCE dynamic contrast enhanced MR urography for RS renal scintigraphy acceptable. For severely dilated kidneys, a mean underestimation of SRF split renal function of 4% should be expected with DCE dynamic contrast enhanced MR urography, making substitution questionable.

Multiparametric optical and MR imaging demonstrate inhibition of tumor angiogenesis natural history by mural cell therapy.

PURPOSE: To determine whether functional imaging using MRI and fibered confocal fluorescence microscopy (FCFM) could be used to monitor cell therapy by mural progenitor cells (MPC). METHODS: Fifty mice bearing TC1 murine xenograft tumors were allocated into: control (n = 17), sham (phosphate buffer saline, n = 16), and MPC-treated (MPC, n = 17) groups. MRI was performed before (D0 ) and 7 days (D7 ) after injection measuring tumor size, R2 * under air, oxygen, and carbogen using blood oxygen level dependent (BOLD) and f (fraction linked to microcirculation), D* (perfusion related coefficient) and Dr (restricted diffusion coefficient) using diffusion-weighted sequences based on the IVIM (intravoxel incoherent motion) method. FCFM was performed at D7 measuring "index leakage" (capillary permeability). RESULTS: Tumor growth was significantly slowed down in the MPC-treated animals (P = 0.002) on D7 . R2 *air significantly decreased in controls between D0 and D7 (P = 0.03), reflecting a decrease in tumor oxygenation. ΔR2 *O2CO2 significantly increased in controls between D0 and D7 (P = 0.01) reflecting loss of vessel response to carbogen. D* significantly decreased in controls between D0 and D7 (P = 0.03). Finally, "index leakage" was lower in the MPC-treated tumors (P = 0,009). CONCLUSION: Treatment by MPC resulted in slowing down of tumor growth, capillary permeability decrease, and stabilization of tumor angiogenesis.

Fetoplacental oxygenation in an intrauterine growth restriction rat model by using blood oxygen level-dependent MR imaging at 4.7 T.

PURPOSE: To investigate blood oxygen level-dependent (BOLD) magnetic resonance (MR) imaging in an intrauterine growth restriction (IUGR) rat model as a noninvasive in vivo tool to evaluate the response of the fetoplacental units (FPUs) to oxygenation MATERIALS AND METHODS: All procedures were approved by the animal care committee. The study was performed between February and July 2010. The IUGR model based on the ligation of the left uterine vascular pedicle at embryonic day 17 of gestation was validated by weighing placentas and fetuses after MR imaging. FPUs in the left and right uterine horns were IUGR cases and controls, respectively. A small-animal 4.7-T MR imager was used. Multiple gradient-echo sequence (repetition time msec/echo time msec, 800/1.8-49.8) was performed at embryonic day 19. T2* relaxation time was measured before and after maternal hyperoxygenation for live FPUs in placenta, fetal liver, and brain. The effect of hyperoxygenation on BOLD MR imaging was analyzed with change in T2* between hyperoxygenation and ambient air. After dissection, live fetuses from both horns were identified and weighed. Changes in T2* were compared based on Student t tests. A mixed model was used to compare BOLD effect among horns and organs. RESULTS: Sixteen rats were studied. There was a significant fetal weight decrease in the IUGR FPUs (-21.9%; P < .001). Change in T2* differed significantly between IUGR cases and controls for placenta (5.25 msec vs 11.25 msec; P < .001) and fetal brain (3.7 msec vs 7.17 msec; P = .02), whereas there was no significant difference in the fetal liver (2.72 msec vs 3.18 msec; P = .47). CONCLUSION: BOLD MR imaging at 4.7 T can be used to evaluate the response to oxygenation in normal and IUGR FPUs. This technique has a potential role in the assessment of human pregnancy.

Evaluation of rat heart microvasculature with high-spatial-resolution susceptibility-weighted MR imaging.

PURPOSE: To demonstrate that strandlike hypointense signals seen in the myocardium of normal rat hearts correspond to myocardial microvessels with high-spatial-resolution susceptibility-weighted (SW) magnetic resonance (MR) imaging without injection of contrast medium. MATERIALS AND METHODS: Animal experiments were performed with institutional animal care committee approval. Ex vivo cardiac MR imaging was performed in 10 normal Wistar rats with a 4.7-T imager and a cryogenic probe. The hypothesis that thin tubular hypointense signals in the myocardium of rat hearts at SW MR imaging sequences (group 1, n = 6; in-plane resolution, 39 µm) represent intramyocardial microvessels was tested. A superparamagnetic intravascular contrast agent (ferumoxsil; Lumirem) was used to explore the distribution of the intramyocardial microvessels (group 2, n = 4; three-dimensional fast imaging with steady-state free precession sequences). Nonparametric Mann-Whitney tests were performed to compare groups 1 and 2 both for microvascular densities (MVD) on histologic sections and for MR imaging signal intensities (SIs). Wilcoxon signed rank tests were used for paired comparison of subepicardial and subendocardial MVD and SI within groups. RESULTS: Ferumoxsil opacified the coronary microvasculature (group 2) on MR-matched histologic sections. No statistically significant difference was found between groups 1 and 2 for either MVD or MR imaging SI expressed as ratios between subendocardium and subepicardium (P = .40 and P = .46, respectively). The comparison of mean subendocardial and subepicardial SI within groups revealed significantly more microvessels in the subepicardium with MR (group 1: P = .01; group 2: P = .004). CONCLUSION: Myocardial microvessels appear as strandlike structures on high-spatial-resolution SW MR images without the aid of contrast medium injection.

Machine Learning of Multi-Modal Tumor Imaging Reveals Trajectories of Response to Precision Treatment.

The standard assessment of response to cancer treatments is based on gross tumor characteristics, such as tumor size or glycolysis, which provide very indirect information about the effect of precision treatments on the pharmacological targets of tumors. Several advanced imaging modalities allow for the visualization of targeted tumor hallmarks. Descriptors extracted from these images can help establishing new classifications of precision treatment response. We propose a machine learning (ML) framework to analyze metabolic-anatomical-vascular imaging features from positron emission tomography, ultrafast Doppler, and computed tomography in a mouse model of paraganglioma undergoing anti-angiogenic treatment with sunitinib. Imaging features from the follow-up of sunitinib-treated (n = 8, imaged once-per-week/6-weeks) and sham-treated (n = 8, imaged once-per-week/3-weeks) mice groups were dimensionally reduced and analyzed with hierarchical clustering Analysis (HCA). The classes extracted from HCA were used with 10 ML classifiers to find a generalized tumor stage prediction model, which was validated with an independent dataset of sunitinib-treated mice. HCA provided three stages of treatment response that were validated using the best-performing ML classifier. The Gaussian naive Bayes classifier showed the best performance, with a training accuracy of 98.7 and an average area under curve of 100. Our results show that metabolic-anatomical-vascular markers allow defining treatment response trajectories that reflect the efficacy of an anti-angiogenic drug on the tumor target hallmark.

Correction: Full-field optical coherence tomography for the diagnosis of giant cell arteritis.

[This corrects the article DOI: 10.1371/journal.pone.0234165.].

Acute stress induces long-term metabolic, functional, and structural remodeling of the heart.

Takotsubo cardiomyopathy is a stress-induced cardiovascular disease with symptoms comparable to those of an acute coronary syndrome but without coronary obstruction. Takotsubo was initially considered spontaneously reversible, but epidemiological studies revealed significant long-term morbidity and mortality, the reason for which is unknown. Here, we show in a female rodent model that a single pharmacological challenge creates a stress-induced cardiomyopathy similar to Takotsubo. The acute response involves changes in blood and tissue biomarkers and in cardiac in vivo imaging acquired with ultrasound, magnetic resonance and positron emission tomography. Longitudinal follow up using in vivo imaging, histochemistry, protein and proteomics analyses evidences a continued metabolic reprogramming of the heart towards metabolic malfunction, eventually leading to irreversible damage in cardiac function and structure. The results combat the supposed reversibility of Takotsubo, point to dysregulation of glucose metabolic pathways as a main cause of long-term cardiac disease and support early therapeutic management of Takotsubo.

Macromolecular capillary leakage is involved in the onset of anaphylactic hypotension.

BACKGROUND: The role of the hypovolemic component secondary to the microcirculatory changes in the onset of inaugural anaphylactic hypotension remains debated. We investigated the microcirculatory permeability in a model of anaphylactic shock using a fluorescence confocal microscopy imaging system. METHODS: Ovalbumin-sensitized anesthetized Brown Norway rats were randomly allocated into two groups (n = 6/group): control and anaphylaxis, respectively induced by intravenous saline or ovalbumin at time 0 (T0). The mesentery was surgically exposed. Macromolecular fluorescein isothiocyanate-dextran was intravenously injected (T0-5min) allowing in vivo visualization of the mesenteric microvascular network by fluorescence microscopy. After a period of stabilization of the contrast agent concentration, a 5-s movie was recorded to obtain baseline signal intensity. Following T0, 5-s movies were recorded every 30 s for 30 min. Capillary leakage of fluorescein isothiocyanate-dextran was assessed in interstitium and compared between groups. Data are expressed as mean ± SD. RESULTS: Following anaphylactic shock onset, an early, progressive, and global signal intensity increase over time was detected in the interstitium. Mean index leakage differed between control and anaphylaxis (respectively 20 ± 11 vs. 170 ± 127%; P < 0.0001), starting at 2 min after shock onset and progressively increasing. Index leakage correlated with the drop in arterial blood pressure until T0 + 10 min (r = -0.75, P = 0.0001). CONCLUSIONS: During anaphylaxis, interstitial capillary leakage occurs within minutes after shock onset. Compared with controls, the mesenteric microcirculation showed at least 8-fold-increased macromolecular capillary leakage. The inflammation-induced microcirculatory changes with subsequent intravascular fluid transfer might be involved in the onset of the inaugural hypotension during anaphylactic shock.