Quantitative MRI imaging of parenchyma and venation networks in Brassica napus leaves: effects of development and dehydration.

BACKGROUND: Characterisation of the structure and water status of leaf tissues is essential to the understanding of leaf hydraulic functioning under optimal and stressed conditions. Magnetic Resonance Imaging is unique in its capacity to access this information in a spatially resolved, non-invasive and non-destructive way. The purpose of this study was to develop an original approach based on transverse relaxation mapping by Magnetic Resonance Imaging for the detection of changes in water status and distribution at cell and tissue levels in Brassica napus leaves during blade development and dehydration. RESULTS: By combining transverse relaxation maps with a classification scheme, we were able to distinguish specific zones of areoles and veins. The tissue heterogeneity observed in young leaves still occurred in mature and senescent leaves, but with different distributions of T2 values in accordance with the basipetal progression of leaf blade development, revealing changes in tissue structure. When subjected to severe water stress, all blade zones showed similar behaviours. CONCLUSION: This study demonstrates the great potential of Magnetic Resonance Imaging in assessing information on the structure and water status of leaves. The feasibility of in planta leaf measurements was demonstrated, opening up many opportunities for the investigation of leaf structure and hydraulic functioning during development and/or in response to abiotic stresses.

Growth kinetics, spatialization and quality of potato tubers monitored in situ by MRI - long-term effects of water stress.

Understanding the potato tuber development and effects of drought at key stages of sensitivity on yield is crucial, particularly when considering the increasing incidence of drought due to climate change. So far, few studies addressed the time course of tuber growth in soil, mainly due to difficulties in accessing underground plant organs in a non-destructive manner. This study aims to understand the tuber growth and quality and the complex long-term effects of realistic water stress on potato tuber yield. MRI was used to monitor the growth kinetics and spatialization of individual tubers in situ and the evolution of internal defects throughout the development period. The intermittent drought applied to plants reduced tuber yield by reducing tuber growth and increasing the number of aborted tubers. The reduction in the size of tubers depended on the vertical position of the tubers in the soil, indicating water exchanges between tubers and the mother plant during leaf dehydration events. The final size of tubers was linked with the growth rate at specific developmental periods. For plants experiencing stress, this corresponded to the days following rewatering, suggesting tuber growth plasticity. All internal defects occurred in large tubers and within a short time span immediately following a period of rapid growth of perimedullary tissues, probably due to high nutrient requirements. To conclude, the non-destructive 3D imaging by MRI allowed us to quantify and better understand the kinetics and spatialization of tuber growth and the appearance of internal defects under different soil water conditions.

Quantitative magnetic resonance imaging of in vitro gastrointestinal digestion of a bread and cheese meal.

The monitoring of food degradation during gastrointestinal digestion is essential in understanding food structure impacts on the bioaccessibility and bioavailability of nutrients. Magnetic Resonance Imaging (MRI) has the unique ability to access information on changes in multi-scale structural features of foods in a spatially resolved and non-destructive way. Our objective was to exploit various opportunities offered by MRI for monitoring starch, lipid and protein hydrolysis, as well as food particle breakdown during the semi-dynamic in vitro gastrointestinal digestion of complex foods combined in a meal. The meal consisted of French bread, hard cheese and water (drink), with a realistic distribution of bolus particle sizes. The MRI approach was reinforced by parallel chemical analysis of all macronutrients in the supernatant. By combining different imaging protocols, quantitative MRI provided insights into a number of phenomena at the level of the cheese and bread particles and within the liquid phase that are hard to access through conventional approaches. MRI thus revealed the progressive ingress of fluids into the bread crust and the release of the gas trapped in the crumb, the erosion of cheese particles, the creaming of fat, the disappearance of small food particles and changes in liquid phase composition. Excellent agreement was obtained between the quantitative parameters extracted from the MRI images and the results of the chemical analysis, demonstrating the strong potential of MRI for the monitoring of in vitro gastrointestinal digestion. The present study proposes further improvements to fully exploit the capabilities of MRI and constitutes an important step towards the extension of quantitative MRI to in vivo studies.

Characterization of Potato Tuber Tissues Using Spatialized MRI T2 Relaxometry.

Magnetic Resonance Imaging is a powerful non-destructive tool in the study of plant tissues. For potato tubers, it greatly assists the study of tissue defects and tissue evolution during storage. This paper describes the MRI analysis of potato tubers with internal defects in their flesh tissue at eight sampling dates from 14 to 33 weeks after harvest. Spatialized multi-exponential T2 relaxometry was used to generate bi-exponential T2 maps, coupled with a classification scheme to identify the different T2 homogeneous zones within the tubers. Six classes with statistically different relaxation parameters were identified at each sampling date, allowing the defects and the pith and cortex tissues to be detected. A further distinction could be made between three constitutive elements within the flesh, revealing the heterogeneity of this particular tissue. Relaxation parameters for each class and their evolution during storage were successfully analyzed. The work demonstrated the value of MRI for detailed non-invasive plant tissue characterization.

Characterization of the Water Shortage Effects on Potato Tuber Tissues during Growth Using MRI Relaxometry and Biochemical Parameters.

The potato is one of the most cultivated crops worldwide, providing an important source of food. The quality of potato tubers relates to their size and dry matter composition and to the absence of physiological defects. It depends on the spatial and temporal coordination of growth and metabolic processes in the major tuber tissues: the cortex, flesh and pith. In the present study, variations in the biochemical traits of each of these tissues were investigated during tuber growth under optimal and water-deficit conditions. MRI relaxometry was used as a non-invasive and quantitative method to access information on cellular water status. The presence of slight but significant variations in organic compound contents quantified in the cortex and flesh revealed a tissue-dependent metabolic pattern. The T2 and relative I0 of the bi-exponential relaxation signal allowed a distinction to be made between the pith and the cortex, whereas the flesh could be differentiated from these tissues only through its relative I0. T2 values did not vary significantly during tuber development, in accordance with the typical growth pattern of tubers, but were shown to be sensitive to water stress. The interpretation of the multi-exponential transverse relaxation times is discussed and could be further developed via microscopic analysis.

Spatial and temporal evolution of quantitative magnetic resonance imaging parameters of peach and apple fruit - relationship with biophysical and metabolic traits.

Fruits are complex organs that are spatially regulated during development. Limited phenotyping capacity at cell and tissue levels is one of the main obstacles to our understanding of the coordinated regulation of the processes involved in fruit growth and quality. In this study, the spatial evolution of biophysical and metabolic traits of peach and apple fruit was investigated during fruit development. In parallel, the multi-exponential relaxation times and apparent microporosity were assessed by quantitative magnetic resonance imaging (MRI). The aim was to identify the possible relationships between MRI parameters and variations in the structure and composition of fruit tissues during development so that transverse relaxation could be proposed as a biomarker for the assessment of the structural and functional evolution of fruit tissues during growth. The study provides species-specific data on developmental and spatial variations in density, cell number and size distribution, insoluble and soluble compound accumulation and osmotic and water potential in the fruit mesocarp. Magnetic resonance imaging was able to capture tissue evolution and the development of pericarp heterogeneity by accessing information on cell expansion, water status and distribution at cell level, and microporosity. Changes in vacuole-related transverse relaxation rates were mostly explained by cell/vacuole size. The impact of cell solute composition, microporosity and membrane permeability on relaxation times is also discussed. The results demonstrate the usefulness of MRI as a tool to phenotype fruits and to access important physiological data during development, including information on spatial variability.

fMRI-Based Brain Responses to Quinine and Sucrose Gustatory Stimulation for Nutrition Research in the Minipig Model: A Proof-of-Concept Study.

The minipig model is of high interest for brain research in nutrition and associated pathologies considering the similarities to human nutritional physiology, brain structures, and functions. In the context of a gustatory stimulation paradigm, fMRI can provide crucial information about the sensory, cognitive, and hedonic integration of exteroceptive stimuli in healthy and pathological nutritional conditions. Our aims were (i) to validate the experimental setup, i.e., fMRI acquisition and SPM-based statistical analysis, with a visual stimulation; (ii) to implement the fMRI procedure in order to map the brain responses to different gustatory stimulations, i.e., sucrose (5%) and quinine (10 mM), and (ii) to investigate the differential effects of potentially aversive (quinine) and appetitive/pleasant (sucrose) oral stimulation on brain responses, especially in the limbic and reward circuits. Six Yucatan minipigs were imaged on an Avanto 1.5-T MRI under isoflurane anesthesia and mechanical ventilation. BOLD signal was recorded during visual or gustatory (artificial saliva, sucrose, or quinine) stimulation with a block paradigm. With the visual stimulation, brain responses were detected in the visual cortex, thus validating our experimental and statistical setup. Quinine and sucrose stimulation promoted different cerebral activation patterns that were concordant, to some extent, to results from human studies. The insular cortex (i.e., gustatory cortex) was activated with both sucrose and quinine, but other regions were specifically activated by one or the other stimulation. Gustatory stimulation combined with fMRI analysis in large animals such as minipigs is a promising approach to investigate the integration of gustatory stimulation in healthy or pathological conditions such as obesity, eating disorders, or dysgeusia. To date, this is the first intent to describe gustatory stimulation in minipigs using fMRI.

MRI investigation of subcellular water compartmentalization and gas distribution in apples.

Water status and distribution at subcellular level in whole apple fruit were evaluated by Magnetic Resonance Imaging (MRI) measurement of the multi-exponential transverse (T2) relaxation of water protons. Apparent microporosity, also estimated by MRI, provided mapping of gas distribution in fruit tissues. Measuring for the first time the multi-exponential relaxation of water and apparent tissue microporosity in whole fruit and combining these with histological measurements provided a more reliable interpretation of the origins of variations in the transverse relaxation time (T2) and better characterization of the fruit tissue. Measurements were performed on 54 fruits from 3 different cultivars. Fruits of different sizes were selected for each cultivar to provide tissues with cells of different dimensions. Macrovision measurements were carried out on parenchymal tissue from all fruits to investigate the impact of cell size on T2 value. The results showed that the MRI transverse relaxation signal is well fitted by a tri-exponential decay curve that reflects cell compartmentalization. Variations in cell size partially explained the different T2 observed. This study highlighted the heterogeneity of apple tissues in terms of relaxation parameters, apparent microporosity and cell morphology and in relation to specific variations between fruit of different cultivars.

MSE-MRI sequence optimisation for measurement of bi- and tri-exponential T2 relaxation in a phantom and fruit.

The transverse relaxation signal from vegetal cells can be described by multi-exponential behaviour, reflecting different water compartments. This multi-exponential relaxation is rarely measured by conventional MRI imaging protocols; mono-exponential relaxation times are measured instead, thus limiting information about of the microstructure and water status in vegetal cells. In this study, an optimised multiple spin echo (MSE) MRI sequence was evaluated for assessment of multi-exponential transverse relaxation in fruit tissues. The sequence was designed for the acquisition of a maximum of 512 echoes. Non-selective refocusing RF pulses were used in combination with balanced crusher gradients for elimination of spurious echoes. The study was performed on a bi-compartmental phantom with known T2 values and on apple and tomato fruit. T2 decays measured in the phantom and fruit were analysed using bi- and tri-exponential fits, respectively. The MRI results were compared with low field non-spatially resolved NMR measurements performed on the same samples. The results demonstrated that the MSE-MRI sequence can be used for up to tri-exponential T2 quantification allowing for estimation of relaxation times from a few tens of milliseconds to over a second. The effects of the crusher moment and the TE value on T2 measurements were studied both on the bi-compartmental phantom and on the fruit tissues. It was demonstrated that the sequence should be optimised with regard to the characteristics of the tissue to be examined by considering the effects of water molecular diffusion in the presence of both imaging gradients and gradients produced by susceptibility inhomogeneities.

Rapid quantification of muscle fat content and subcutaneous adipose tissue in fish using MRI.

The potentiality of MRI to quantify fat content in flesh and subcutaneous fat in fish cutlets was investigated. Low measurement time was aimed at in a view to handling large number of samples needed in selective breeding programs for example. Results on fresh and frozen-thawed cutlets were compared to assess this way of conservation. As MRI generates unwanted spatial variations of the signal, a correction method was developed enabling the measurement on several cutlets simultaneously in less than 3 min per sample. For subcutaneous fat, the results were compared with vision measurements. High correlations between both techniques were found (R(2)=0.77 and 0.87 for the ventral and dorsal part). Fat in flesh was validated vs NMR measurements. No statistical difference was found between fresh and frozen-thawed cutlets. RMSE was respectively 0.8% and 0.89%. These results confirmed the potentiality of MRI for fat measurement in fish particularly for a large number of samples.

Temperature mapping in bread dough using SE and GE two-point MRI methods: experimental and theoretical estimation of uncertainty.

Two-dimensional (2D)-SE, 2D-GE and tri-dimensional (3D)-GE two-point T(1)-weighted MRI methods were evaluated in this study in order to maximize the accuracy of temperature mapping of bread dough during thermal processing. Uncertainties were propagated throughout each protocol of measurement, and comparisons demonstrated that all the methods with comparable acquisition times minimized the temperature uncertainty to similar extent. The experimental uncertainties obtained with low-field MRI were also compared to the theoretical estimations. Some discrepancies were reported between experimental and theoretical values of uncertainties of temperature; however, experimental and theoretical trends with varying parameters agreed to a large extent for both SE and GE methods. The 2D-SE method was chosen for further applications on prefermented dough because of its lower sensitivity to susceptibility differences in porous media. It was applied for temperature mapping in prefermented dough during chilling prior to freezing and compared locally to optical fiber measurements.

Quantification of microporosity in fruit by MRI at various magnetic fields: comparison with X-ray microtomography.

Microstructure determines the mechanical and transport properties of fruit tissues. One important characteristic of the microstructure is the relative volume fraction of gas-filled intercellular spaces, i.e., the tissue microporosity. Quantification of this microporosity is fundamental for investigating the relationship between gas transfer and various disorders in fruit. We present a new method for quantifying the apparent microporosity using magnetic resonance imaging (MRI). The method is based on the differences in magnetic susceptibility between gas-filled intercellular spaces and their environment inside fruit tissues. It was tested at two different magnetic fields (1.5 and 0.2 T) on apple and tomato fruit. The method was validated by comparing the MRI results with estimation of local tissue porosity using X-ray microtomography experiments. MRI was shown to be effective in determining the distribution of apparent microporosity in fruit.

An investigation of the structural aspects of the tomato fruit by means of quantitative nuclear magnetic resonance imaging.

In this study, magnetic resonance imaging (MRI) was applied to study the structural aspects of the tomato fruit. The main study was performed on tomatoes (cv. Tradiro) using a 0.2-T electromagnet scanner. Spin-echo images were acquired to visualize the tomato macrostructure. The air bubble content in tissues was evaluated by exploiting susceptibility effects using multiple gradient echo images. The microstructure was further studied by measuring spin-spin (T(2)) and spin-lattice (T(1)) relaxation time distributions. Nuclear magnetic resonance relaxometry, macro vision imaging and chemical analysis were used as complementary and independent experimental methods in order to emphasize the MRI results. MRI images showed that the air bubble content varied between tissues. The presence of gas was attested by macro vision images. Quantitative imaging showed that T(2) and T(1) maps obtained by MRI reflected the structural differences between tomato tissues and made it possible to distinguish between them. The results indicated that cell size and chemical composition contribute to the relaxation mechanism.

Noninvasive characterization of gonad maturation and determination of the sex of Pacific oysters by MRI.

The aim of this study was to test the ability of magnetic resonance imaging (MRI) technique to characterize gonad development and to determine the sex of live Pacific oysters through their shells. A preliminary nuclear magnetic resonance (NMR) relaxometry study was conducted to characterize T1 and T2 NMR relaxation parameters for the main oyster organs. This showed that T1-weighted MRI sequences were most appropriate to optimize contrasts between tissues in images. The results showed that gray levels of gonads in images acquired with gradient-echo sequence were variably affected by T2* weighting effect. However, the ovaries systematically gave a hypersignal in spin-echo T1-weighted images, and stack histograms of female oysters showed a peak well separated from that of male oysters. An automated method is proposed to quantify the development of oysters and their gonad maturation and to identify their sex.