The four-minute approach revisited: accelerating MRI-based multi-factorial age estimation.

OBJECTIVES: This feasibility study aimed to investigate the reliability of multi-factorial age estimation based on MR data of the hand, wisdom teeth and the clavicles with reduced acquisition time. METHODS: The raw MR data of 34 volunteers-acquired on a 3T system and using acquisition times (TA) of 3:46 min (hand), 5:29 min (clavicles) and 10:46 min (teeth)-were retrospectively undersampled applying the commercially available CAIPIRINHA technique. Automatic and radiological age estimation methods were applied to the original image data as well as undersampled data to investigate the reliability of age estimates with decreasing acquisition time. Reliability was investigated determining standard deviation (SSD) and mean (MSD) of signed differences, intra-class correlation (ICC) and by performing Bland-Altman analysis. RESULTS: Automatic age estimation generally showed very high reliability (SSD < 0.90 years) even for very short acquisition times (SSD ≈ 0.20 years for a total TA of 4 min). Radiological age estimation provided highly reliable results for images of the hand (ICC ≥ 0.96) and the teeth (ICC ≥ 0.79) for short acquisition times (TA = 16 s for the hand, TA = 2:21 min for the teeth), imaging data of the clavicles allowed for moderate acceleration (TA = 1:25 min, ICC ≥ 0.71). CONCLUSIONS: The results demonstrate that reliable multi-factorial age estimation based on MRI of the hand, wisdom teeth and the clavicles can be performed using images acquired with a total acquisition time of 4 min.

Reducing acquisition time for MRI-based forensic age estimation.

Radiology-based estimation of a living person's unknown age has recently attracted increasing attention due to large numbers of undocumented immigrants entering Europe. To avoid the application of X-ray-based imaging techniques, magnetic resonance imaging (MRI) has been suggested as an alternative imaging modality. Unfortunately, MRI requires prolonged acquisition times, which potentially represents an additional stressor for young refugees. To eliminate this shortcoming, we investigated the degree of reduction in acquisition time that still led to reliable age estimates. Two radiologists randomly assessed original images and two sets of retrospectively undersampled data of 15 volunteers (N = 45 data sets) applying an established radiological age estimation method to images of the hand and wrist. Additionally, a neural network-based age estimation method analyzed four sets of further undersampled images from the 15 volunteers (N = 105 data sets). Furthermore, we compared retrospectively undersampled and acquired undersampled data for three volunteers. To assess reliability with increasing degree of undersampling, intra-rater and inter-rater agreement were analyzed computing signed differences and intra-class correlation. While our findings have to be confirmed by a larger prospective study, the results from both radiological and automatic age estimation showed that reliable age estimation was still possible for acquisition times of 15 seconds.

Reproducibility of relaxometry of human lumbar vertebrae at 3 Tesla using 1 H MR spectroscopy.

BACKGROUND: MR spectroscopy is widely used for fat fraction quantification of human lumbar vertebrae. However, the measurements need to be corrected for relaxation effects. PURPOSE: The aim of this study was to determine the reproducibility of relaxometry in human lumbar vertebrae required for the correction of fat fraction measurements using magnetic resonance spectroscopy at 3 Tesla. Such information provides error estimates and guidance regarding reliability for future studies. STUDY TYPE: Prospective. SUBJECTS: Forty-six healthy volunteers (22 female [f], 24 male [m]) participated in this study. FIELD STRENGTH: All subjects underwent three consecutive multi-TE /multi-TR MR spectroscopy measurements at 3 Tesla. ASSESSMENT: A total of 2580 spectra of lumbar vertebrae L2 and L3 of 43 subjects (21f, 22m) were quantified using jMRUI software. Data were exported and mono-exponential fits were applied to the signals of water and fat compartments to derive relaxation times and calculate the fat fraction corrected for relaxation effects. Finally, relaxation times and fat fraction results of repeated measurements were analyzed for reproducibility. STATISTICAL TESTS: Reproducibility was evaluated by calculating the coefficient of variation (CV). Influences of volunteer age and sex were tested by analysis of covariance. RESULTS: The CV for all calculated parameters ranged between 1.22% (T2 of the fat compartment) and 3.02% (T1 of the fat compartment). Relaxation times and fat fraction were statistically different for female and male volunteers (P < 0.01) and relaxation times of the water compartment showed significant (P < 0.01) correlation with the fat fraction. DATA CONCLUSION: Based on repeated acquisitions using the measurement parameters applied in this study, magnetic resonance spectroscopy allowed a reproducible calculation of the fat fraction corrected for relaxation effects. T1 of the water compartment showed high reproducibility and correlation with the fat fraction. It, therefore, might be considered as a parameter linked to the composition of the water compartment and patient health. LEVEL OF EVIDENCE: 1 Technical Efficacy Stage 1 J. Magn. Reson. Imaging 2017.

Forensic age estimation by morphometric analysis of the manubrium from 3D MR images.

Forensic age estimation research based on skeletal structures focuses on patterns of growth and development using different bones. In this work, our aim was to study growth-related evolution of the manubrium in living adolescents and young adults using magnetic resonance imaging (MRI), which is an image acquisition modality that does not involve ionizing radiation. In a first step, individual manubrium and subject features were correlated with age, which confirmed a statistically significant change of manubrium volume (Mvol:p<0.01, R2¯=0.50) and surface area (Msur:p<0.01, R2¯=0.53) for the studied age range. Additionally, shapes of the manubria were for the first time investigated using principal component analysis. The decomposition of the data in principal components allowed to analyse the contribution of each component to total shape variation. With 13 principal components, ∼96% of shape variation could be described (Mshp:p<0.01, R2¯=0.60). Multiple linear regression analysis modelled the relationship between the statistically best correlated variables and age. Models including manubrium shape, volume or surface area divided by the height of the subject (Y∼MshpMsur/Sh:p<0.01, R2¯=0.71; Y∼MshpMvol/Sh:p<0.01, R2¯=0.72) presented a standard error of estimate of two years. In order to estimate the accuracy of these two manubrium-based age estimation models, cross validation experiments predicting age on held-out test sets were performed. Median absolute difference of predicted and known chronological age was 1.18 years for the best performing model (Y∼MshpMsur/Sh:p<0.01, Rp2=0.67). In conclusion, despite limitations in determining legal majority age, manubrium morphometry analysis presented statistically significant results for skeletal age estimation, which indicates that this bone structure may be considered as a new candidate in multi-factorial MRI-based age estimation.

Assessment of pharmacokinetics for microvessel proliferation by DCE-MRI for early detection of physeal bone bridge formation in an animal model.

OBJECTIVES: Bone bridge formation occurs after physeal lesions and can lead to growth arrest if not reversed. Previous investigations on the underlying mechanisms of this formation used histological methods. Therefore, this study aimed to apply a minimally invasive method using dynamic contrast-enhanced MRI (DCE-MRI). MATERIALS AND METHODS: Changes in functional parameters related to the microvessel system were assessed in a longitudinal study of a cohort of an animal model applying a reference region model. The development of morphology of the injured physis was investigated with 3D high-resolution MRI. To acquire complementary information for MRI-related findings qRT-PCR and immunohistochemical data were acquired for a second cohort of the animal model. RESULTS: The evaluation of the pharmacokinetic parameters showed a first rise of the transfer coefficient 7 days post-lesion and a maximum 42 days after operation. The analysis of the complementary data showed a connection of the first rise to microvessel proliferation while the maximum value was linked to bone remodeling. CONCLUSION: The pharmacokinetic analysis of DCE-MRI provides information on a proliferation of microvessels during the healing process as a sign for bone bridge formation. Thereby, DCE-MRI could identify details, which up to now required analyses of highly invasive methods.

Temperature dependence of viscosity, relaxation times (T1, T2) and simulated contrast for potential perfusates in post-mortem MR angiography (PMMRA).

Developments in post-mortem imaging increasingly focus on addressing recognised diagnostic weaknesses, especially with regard to suspected natural deaths. Post-mortem MR angiography (PMMRA) may offer additional diagnostic information to help address such weaknesses, specifically in the context of sudden cardiac death. Complete filling of the coronary arteries and acceptable contrast with surrounding tissue are essential for a successful approach to PMMRA. In this work, the suitability of different liquids for inclusion in a targeted PMMRA protocol was evaluated. Factors influencing cooling of paraffinum liquidum + Angiofil® (6 %) in cadavers during routine multiphase post-mortem CT angiography were investigated. The temperature dependence of dynamic viscosity (8-20 °C), longitudinal (T1) and transverse (T2) relaxation (1-23 °C) of the proposed liquids was quadratically modelled. The relaxation behaviour of these liquids and MR scan parameters were further investigated by simulation of a radiofrequency (RF)-spoiled gradient echo (GRE) sequence to estimate potentially achievable contrast between liquids and post-mortem tissue at different temperatures across a forensically relevant temperature range. Analysis of the established models and simulations indicated that based on dynamic viscosity (27-33 mPa · s), short T1 relaxation times (155-207 ms) and a minimal temperature dependence over the investigated range of these parameters, paraffin oil and a solution of paraffin oil + Angiofil® (6 %) would be most suitable for post-mortem reperfusion and examination in MRI.

Time-Dependent Changes in T1 during Fracture Healing in Juvenile Rats: A Quantitative MR Approach.

Quantitative magnetic resonance imaging (qMRI) offers several advantages in imaging and determination of soft tissue alterations when compared to qualitative imaging techniques. Although applications in brain and muscle tissues are well studied, its suitability to quantify relaxation times of intact and injured bone tissue, especially in children, is widely unknown. The objective observation of a fracture including its age determination can become of legal interest in cases of child abuse or maltreatment. Therefore, the aim of this study is the determination of time dependent changes in intact and corresponding injured bones in immature rats via qMRI, to provide the basis for an objective and radiation-free approach for fracture dating. Thirty-five MR scans of 7 Sprague-Dawley rats (male, 4 weeks old, 100 ± 5 g) were acquired on a 3T MRI scanner (TimTrio, Siemens AG, Erlangen, Germany) after the surgical infliction of an epiphyseal fracture in the tibia. The images were taken at days 1, 3, 7, 14, 28, 42 and 82 post-surgery. A proton density-weighted and a T1-weighted 3D FLASH sequence were acquired to calculate the longitudinal relaxation time T1 of the fractured region and the surrounding tissues. The calculation of T1 in intact and injured bone resulted in a quantitative observation of bone development in intact juvenile tibiae as well as the bone healing process in the injured tibiae. In both areas, T1 decreased over time. To evaluate the differences in T1 behaviour between the intact and injured bone, the relative T1 values (bone-fracture) were calculated, showing clear detectable alterations of T1 after fracture occurrence. These results indicate that qMRI has a high potential not only for clinically relevant applications to detect growth defects or developmental alterations in juvenile bones, but also for forensically relevant applications such as the dating of fractures in cases of child abuse or maltreatment.

Quantitative MR imaging in fracture dating--Initial results.

For exact age determinations of bone fractures in a forensic context (e.g. in cases of child abuse) improved knowledge of the time course of the healing process and use of non-invasive modern imaging technology is of high importance. To date, fracture dating is based on radiographic methods by determining the callus status and thereby relying on an expert's experience. As a novel approach, this study aims to investigate the applicability of magnetic resonance imaging (MRI) for bone fracture dating by systematically investigating time-resolved changes in quantitative MR characteristics after a fracture event. Prior to investigating fracture healing in children, adults were examined for this study in order to test the methodology for this application. Altogether, 31 MR examinations in 17 subjects (♀: 11 ♂: 6; median age 34 ± 15 y, scanned 1-5 times over a period of up to 200 days after the fracture event) were performed on a clinical 3T MR scanner (TimTrio, Siemens AG, Germany). All subjects were treated conservatively for a fracture in either a long bone or in the collar bone. Both, qualitative and quantitative MR measurements were performed in all subjects. MR sequences for a quantitative measurement of relaxation times T1 and T2 in the fracture gap and musculature were applied. Maps of quantitative MR parameters T1, T2, and magnetisation transfer ratio (MTR) were calculated and evaluated by investigating changes over time in the fractured area by defined ROIs. Additionally, muscle areas were examined as reference regions to validate this approach. Quantitative evaluation of 23 MR data sets (12 test subjects, ♀: 7 ♂: 5) showed an initial peak in T1 values in the fractured area (T1=1895 ± 607 ms), which decreased over time to a value of 1094 ± 182 ms (200 days after the fracture event). T2 values also peaked for early-stage fractures (T2=115 ± 80 ms) and decreased to 73 ± 33 ms within 21 days after the fracture event. After that time point, no significant changes could be detected for T2. MTR remained constant at 35.5 ± 8.0% over time. The study shows that the quantitative assessment of T1 and T2 behaviour over time in the fractured region enable the generation of a novel model allowing for an objective age determination of a fracture.

Positive contrast of SPIO-labeled cells by off-resonant reconstruction of 3D radial half-echo bSSFP.

This article describes a new acquisition and reconstruction concept for positive contrast imaging of cells labeled with superparamagnetic iron oxides (SPIOs). Overcoming the limitations of a negative contrast representation as gained with gradient echo and fully balanced steady state (bSSFP), the proposed method delivers a spatially localized contrast with high cellular sensitivity not accomplished by other positive contrast methods. Employing a 3D radial bSSFP pulse sequence with half-echo sampling, positive cellular contrast is gained by adding artificial global frequency offsets to each half-echo before image reconstruction. The new contrast regime is highlighted with numerical intravoxel simulations including the point-spread function for 3D half-echo acquisitions. Furthermore, the new method is validated on the basis of in vitro cell phantom measurements on a clinical MRI platform, where the measured contrast-to-noise ratio (CNR) of the new approach exceeds even the negative contrast of bSSFP. Finally, an in vivo proof of principle study based on a mouse model with a clear depiction of labeled cells within a subcutaneous cell islet containing a cell density as low as 7 cells/mm(3) is presented. The resultant isotropic images show robustness to motion and a high CNR, in addition to an enhanced specificity due to the positive contrast of SPIO-labeled cells.

Contrast of artificial subcutaneous hematomas in MRI over time.

In clinical forensic medicine, hematomas and other externally visible injuries build the basis for the reconstruction of events. However, dating of subcutaneous hematomas based on their external aspect is difficult. Magnetic resonance imaging (MRI) has proven its use in dating intracranial hemorrhage. Thus, the aim was to investigate if MRI can also be used for dating subcutaneous hematomas and to analyze an eventual influence of the hematoma shape. In 20 healthy volunteers (11 females, 9 males, aged 26.9 ± 3.8 years), 4 ml of autologous blood were injected subcutaneously in the thigh. The hematoma was scanned immediately after the injection, after 3 and 24 h and 3, 7, and 14 days using three sequences with different contrast. Data was analyzed by measuring signal intensities of the hematoma, the muscle, and the subcutaneous tissue over time, and the Michelson contrast coefficients between the tissues were calculated. In the analysis, hematoma shape was considered. Signal intensity of blood in the proton density-weighted sequence reached its maximum 3 h after the injection with a subsequent decrease, whereas the signal intensities of muscle and fatty tissue remained constant. The time course of the Michelson coefficient of blood versus muscle decreased exponentially with a change from hyperintensity to hypointensity at 116.9 h, depending on hematoma shape. In the other sequences, either variability was large or contrast coefficients stayed constant over time. The observed change of contrast of blood versus muscle permits a quick estimate of a hematoma's age. The consideration of the hematoma shape is expected to further enhance dating using MRI.

Age determination of soft tissue hematomas.

In clinical forensic medicine, the estimation of the age of injuries such as externally visible subcutaneous hematomas is important for the reconstruction of violent events, particularly to include or exclude potential suspects. Since the estimation of the time of origin based on external inspection is unreliable, the aim of this study was to use contrast in MRI to develop an easy-to-use model for hematoma age estimation. In a longitudinal study, artificially created subcutaneous hematomas were repetitively imaged using MRI over a period of two weeks. The hemorrhages were created by injecting autologous blood into the subcutaneous tissue of the thigh in 20 healthy volunteers. For MRI, standard commercially available sequences, namely proton-density-weighted, T2 -weighted and inversion recovery sequences, were used. The hematomas' MRI data were analyzed regarding their contrast behavior using the most suitable sequences to derive a model allowing an objective estimation of the age of soft tissue hematomas. The Michelson contrast between hematoma and muscle in the proton-density-weighted sequence showed an exponentially decreasing behavior with a dynamic range of 0.6 and a maximum standard deviation of 0.1. The contrast of the inversion recovery sequences showed increasing characteristics and was hypointense for TI = 200ms and hyperintense for TI =1000ms. These sequences were used to create a contrast model. The cross-validation of the model finally yielded limits of agreement for hematoma age determination (corresponding to ±1.96 SD) of ±38.7h during the first three days and ±54 h for the entire investigation period. The developed model provides lookup tables which allow for the estimation of a hematoma's age given a single contrast measurement applicable by a radiologist or a forensic physician. This is a first step towards an accurate and objective dating method for subcutaneous hematomas, which will be particularly useful in child abuse.

Fungi treated with small chemicals exhibit increased antimicrobial activity against facultative bacterial and yeast pathogens.

For decades, fungi have been the main source for the discovery of novel antimicrobial drugs. Recent sequencing efforts revealed a still high number of so far unknown "cryptic" secondary metabolites. The production of these metabolites is presumably epigenetically silenced under standard laboratory conditions. In this study, we investigated the effect of six small mass chemicals, of which some are known to act as epigenetic modulators, on the production of antimicrobial compounds in 54 spore forming fungi. The antimicrobial effect of fungal samples was tested against clinically facultative pathogens and multiresistant clinical isolates. In total, 30 samples of treated fungi belonging to six different genera reduced significantly growth of different test organisms compared to the untreated fungal sample (growth log reduction 0.3-4.3). For instance, the pellet of Penicillium restrictum grown in the presence of butyrate revealed significant higher antimicrobial activity against Staphylococcus (S.) aureus and multiresistant S. aureus strains and displayed no cytotoxicity against human cells, thus making it an ideal candidate for antimicrobial compound discovery. Our study shows that every presumable fungus, even well described fungi, has the potential to produce novel antimicrobial compounds and that our approach is capable of rapidly filling the pipeline for yet undiscovered antimicrobial substances.

Laryngeal electromyography: electrode guidance based on 3-dimensional magnetic resonance tomography images of the larynx.

OBJECTIVES/HYPOTHESIS: Laryngeal electromyography (LEMG) is an important tool for the assessment of laryngeal nerve and muscle functioning. The purpose of the study was to determine electrode insertion angle and insertion depth for the various laryngeal muscles. METHODS: Twenty-three cadaver larynges were examined with magnetic resonance tomography (MRT) and Materialize Interactive Medical Image Control System (Leuven, Belgium) 3-dimensional (3D) imaging software. Geometrical analysis was used to calculate the electrode insertion angles. RESULTS: All laryngeal muscles could be identified and 3D visualized on MRT scans. Although the insertion angles were the same in male and female larynges, the insertion depth was significantly larger in male than in female larynges (P<0.05). Of particular clinical importance is the fact that the electrode has to be directed lateral and upward for the thyroarytenoid muscle but lateral and downward for the lateral cricoarytenoid muscle (insertion point=midline lower border of the thyroid). CONCLUSIONS: This is the first study that analyzes electrode insertion angles and insertion depths for each laryngeal muscle using 3D imaging. We hope that the information gained from this study will help clinicians performing LEMG to localize the individual laryngeal muscles.

The spatial and temporal expression of VEGF and its receptors 1 and 2 in post-traumatic bone bridge formation of the growth plate.

Injuries to growth plates may initiate the formation of reversible or irreversible bone-bridges, may leading to bone length discrepancy or axis deviation. As vascular invasion is essential for the formation of bone tissue, the aim of our study was to investigate the kinetic expression of Vascular Endothelial Growth Factor (VEGF) and its receptors R1 and R2 and the ingrowth of vessels in the formation of bone bridges in a rat physeal injury model. Quantitative Real-Time Polymerase Chain Reaction was performed for VEGF and its receptors. Samples from the proximal physis of the tibial bone were immunohistochemically evaluated for the expression of VEGF and its R1 and R2 receptors and Laminin. Morphologically, physeal bone bridge formation was validated by means of Magnetic Resonance Imaging. Kinetic expression of VEGF and VEGF-R1 mRNA documented a tendency towards an increase in expression on day 7. Histological analyses showed a hematoma containing bone debris on day 1 which was replaced with bony trabeculae by day 14, forming a bone bridge by day 28 which was preceded and accompanied by angiogenesis and consistent with MRI data. VEGF and VEGF-R2 was expressed on the debris within the hematoma and bone trabeculae from days 1 to 28. VEGF-R1 expression was only noted until day 14. The findings of our study suggest that physeal bone bridge formation is in part triggered by VEGF expression and associated with angiogenesis, which was shown to precede bone bridge formation and may be further stimulated through VEGF-positive bone debris.