Longitudinal visibility of MRI findings in living victims of strangulation.

Initial experiences with magnetic resonance imaging (MRI) of living strangulation victims demonstrated additional findings of internal injuries compared to the standard clinical forensic examination. However, existing studies on the use of MRI for this purpose mostly focused on the first 48 h after the incident. The aims of this study were (a) to evaluate the longitudinal visibility of MRI findings after violence against the neck by performing two MRI examinations within 12 days and a minimum of four days between both MRI scans and (b) to assess which MRI sequences were most helpful for the detection of injuries. Twenty strangulation victims participated in this study and underwent one (n = 8) or two (n = 12) MRI scans. The first MRI examination was conducted during the first five days, the second five to 12 days after the incident. Two blinded radiologists assessed the MRI data and looked for lesions in the structures of the neck. In total, 140 findings were reported in the 32 MRI examinations. Most of the findings were detected in the thyroid and the muscles of the neck. T2-weighted SPACE with fat suppression, T1-weighted TSE and T1-weighted MPRAGE were rated as the most helpful MRI sequences. Subjects who showed findings in the initial scan also demonstrated comparable results in the second scan, which was performed on average 8.4 days after the incident. Our results show that even up to 12 days after the incident, the criminal proceeding of strangulation cases may greatly profit from the information provided by an MRI examination of the neck in addition to the standard clinical forensic examination.

Temperature correction of post mortem quantitative magnetic resonance imaging using real-time forehead temperature acquisitions.

Performing magnetic resonance imaging (MRI) of deceased is challenging due to altered body temperatures compared to in vivo temperatures and, hence, requires a temperature correction. This study investigates the possibility to correct brain MRI parameters real-time and non invasively based on the forehead temperature. 17 post mortem cases were included and their forehead temperatures were measured continuously during the in situ brain MRI protocol consisting of a diffusion tensor imaging, multi-contrast spin echo, multi-echo gradient echo and inversion recovery spin echo sequence. Linear models were fitted to the quantitative MRI parameters in a forensically interesting temperature range for white matter, cerebral cortex and deep gray matter, separately, and the influence of the forehead temperature on the MRI parameters was determined. A statistically significant temperature sensitivity was found for T2 and mean diffusivity in white matter, for T1 in cerebral cortex, as well as for T1 and mean diffusivity in deep gray matter. Linear models were computed to temperature correct these MRI parameters in in situ post mortem scans to allow their comparison regardless of temperature. The here presented real-time and non invasive temperature correction method for the brain presents a crucial precondition for quantitative in situ post mortem MRI.

Investigation of post mortem brain, rectal and forehead temperature relations.

It is well known that magnetic resonance (MR) imaging is temperature sensitive, which is highly relevant for post mortem examinations. Therefore, the determination of the exact temperature of the investigated body site, e.g. the brain, is crucial. However, direct temperature measurements are invasive and inconvenient. Thus, in view of post mortem MR imaging of the brain, this study aims at investigating the relation between the brain and the forehead temperature for modelling the brain temperature based on the non-invasive forehead temperature. In addition, the brain temperature will be compared to the rectal temperature. Brain temperature profiles measured in the longitudinal fissure between the brain hemispheres, as well as rectal and forehead temperature profiles of 16 deceased were acquired continuously. Linear mixed, linear, quadratic and cubic models were fitted to the relation between the longitudinal fissure and the forehead and between the longitudinal fissure and the rectal temperature, respectively. Highest adjusted R2 values were found between the longitudinal fissure and the forehead temperature, as well as between the longitudinal fissure and the rectal temperature using a linear mixed model including the sex, environmental temperature and humidity as fixed effects. The results indicate that the forehead, as well as the rectal temperature, can be used to model the brain temperature measured in the longitudinal fissure. Comparable fit results were observed for the longitudinal fissure-forehead temperature relation and for the longitudinal fissure-rectal temperature relation. Combined with the fact that the forehead temperature overcomes the problem of measurement invasiveness, the results suggest using the forehead temperature for modelling the brain temperature in the longitudinal fissure.

Technical note: Quantitative optimization of the FLAIR sequence in post mortem magnetic resonance imaging.

The fluid attenuated inversion recovery (FLAIR) magnetic resonance imaging (MRI) sequence aims at suppressing the signal of the cerebrospinal fluid (CSF) by acquiring images at the time point at which the longitudinal magnetization and therefore the signal of CSF is zero. This time point is also called the null point inversion time (TInull). However, the FLAIR sequence is impaired by the temperature dependency of TInull in post mortem MRI due to the lower body temperature of the deceased subject. Therefore, the temperature correction of TInull is crucial for correctly suppressing the CSF signal in post mortem FLAIR imaging. Thus, the goal of this study was to determine the temperature effect on post mortem TInull for achieving a robust suppression of the CSF signal in in situ post mortem MRI using the FLAIR sequence. For this purpose, nine deceased subjects underwent an in situ MRI brain examination on a 3 T MRI scanner. TInull of CSF was determined quantitatively based on different FLAIR acquisitions using varying inversion times. The brain and rectal temperatures were determined prior to the MRI scan. A significant positive linear relation was found between TInull of CSF and the brain temperature, as well as between TInull of CSF and the rectal temperature. The found linear relations between TInull and both brain and rectal temperatures allow correcting TInull for varying temperatures of the deceased. This in turn enables an optimal suppression of the CSF signal in future post mortem FLAIR MRI acquisitions.

Post mortem evaluation of brain edema using quantitative MRI.

The post mortem assessment of brain edema is routinely performed during autopsy by forensic pathologists who evaluate the macroscopically visible signs. In this study, the suitability of magnetic resonance imaging (MRI) for a differentiation between edematous and nonedematous brains was examined as an objective, noninvasive and quantitative rating method. In this study, 22 deceased underwent post mortem in situ MRI prior to regular autopsy. Acquired MRI sequences allowed the computation of the quantitative MRI parameters T1, T2, T2*, fractional anisotropy (FA) and mean diffusivity for the cortex, white matter and deep gray matter separately. Beside the autopsy results, which represent the gold standard for rating brain edema, also the normalized cerebral weight (NCW) was determined by dividing the brain weight by the intracranial volume as developed by Bauer et al. [10]. For further examination of the relation of MRI parameters with the NCW, linear regression models were calculated. The results revealed highly significant correlations of the MRI parameters T2 and FA with the NCW in the cortex. These combinations additionally showed the best fitting results of the linear regression models. In conclusion, quantitative MRI is suitable for differentiating edematous from nonedematous brains by calculating T2 and FA in the cortex. A post mortem in situ MRI scan of the brain can, besides delivering morphological information, add relevant and objective information on the edema status of the brain prior to autopsy or when no autopsy is ordered.

Post mortem brain temperature and its influence on quantitative MRI of the brain.

OBJECTIVE: MRI temperature sensitivity presents a major issue in in situ post mortem MRI (PMMRI), as the tissue temperatures differ from living persons due to passive cooling of the deceased. This study aims at computing brain temperature effects on the MRI parameters to correct for temperature in PMMRI, laying the foundation for future projects on post mortem validation of in vivo MRI techniques. MATERIALS AND METHODS: Brain MRI parameters were assessed in vivo and in situ post mortem using a 3 T MRI scanner. Post mortem brain temperature was measured in situ transethmoidally. The temperature effect was computed by fitting a linear model to the MRI parameters and the corresponding brain temperature. RESULTS: Linear positive temperature correlations were observed for T1, T2* and mean diffusivity in all tissue types. A significant negative correlation was observed for T2 in white matter. Fractional anisotropy revealed significant correlations in all gray matter regions except for the thalamus. DISCUSSION: The linear models will allow to correct for temperature in post mortem MRI. Comparing in vivo to post mortem conditions, the mean diffusivity, in contrast to T1 and T2, revealed additional effects besides temperature, such as cessation of perfusion and active diffusion.

Sensitivity of fiber orientation dependent R2∗ to temperature and post mortem interval.

PURPOSE: R2∗ imaging of brain white matter is well known for being sensitive to the orientation of nerve fibers with respect to the B0 field of the MRI scanner. The goal of this study was to evaluate whether and to which extent fiber orientation dependent R2∗ differs between in vivo and post mortem in situ examinations, and to investigate the influence of varying temperatures and post mortem intervals (PMI). METHODS: Post mortem in situ and in vivo MRI scans were conducted at 3T. R2∗ was acquired with a multi-echo gradient-echo sequence, and the orientation of white matter fibers was computed using diffusion tensor imaging (DTI). Fitting of the measured fiber orientation dependent R2∗ was performed using three different formulations of a previously proposed model. RESULTS: R2∗ increased with increasing fiber angle for in vivo and post mortem in situ examinations, whereby the orientation dependency was lower post mortem. The different formulations of the fiber orientation model resulted in an identical fit, but showed large variations of the estimated parameters. The higher order orientation dependent R2∗ components significantly decreased with decreasing temperature, while the orientation independent R2∗ components showed no significant correlation with either temperature or PMI. CONCLUSION: Although the mean diffusivity is strongly reduced post mortem, we could successfully estimate the fiber angle using DTI. Due to the strong correlation of the higher order orientation dependent R2∗ components with temperature, the decreased R2∗ fiber orientation dependency post mortem in situ might primarily be attributed to the lower brain temperature.

Microcystin ecotypes in a perennial Planktothrix agardhii bloom.

The dynamics and microcystins (MC) concentrations of a perennial Planktothrix agardhii bloom were investigated in a eutrophic lake (Viry-Châtillon, France). A weak relationship was observed between P. agardhii population biomass and the MC concentrations in a 1-year survey. To further investigate the causes of MC concentration changes, we concurrently conducted experiments on 41 strains isolated from this lake. We first checked the clonal diversity of P. agardhii population (i) by molecular techniques, to assess the presence of MC synthetase gene (mcyB), (ii) by biochemical assay (PP2A inhibition assay), for MC production, and (iii) by mass spectrometry (MS), to identify the MC chemotypes. Our results illustrated the diversity of genotype and MC chemotypes within a P. agardhii natural population. Eleven chemotypes among the 16 possible ones were found by MS. Furthermore, we noticed major differences in the MC content of isolated strains (from 0.02 to 1.86 microg equiv. MC-LR mg DW(-1), n=25). Growth and MC production of one MC-producing strain and one non-MC-producing strain were also assessed at two temperatures (10 and 20 degrees C). We showed that growth capacities of these strains were similar at the two tested temperatures, and that the MC production rate was correlated to the growth rate for the MC-producing strain. On the basis of these results, several hypotheses are discussed to explain the weakness of relationships between natural P. agardhii biomass and MC concentration. One of the main reasons could lie in the proportion of MC-producing clones and non-MC-producing clones that may change during the sampling period. Also, the MC-producing clones may present different intracellular MC content due to (i) MC chemotypes diversity, (ii) changes in MC variants proportions within a strain, and (iii) changes in MC rate production depending on the physiological state of cells. Finally, we concluded that various biological organization levels have to be considered (population, cellular and molecular), through an integrative approach, in order to provide a better understanding of P. agardhii in situ MC production.

Seasonal dynamics and toxicity of Cylindrospermopsis raciborskii in Lake Guiers (Senegal, West Africa).

Cylindrospermopsis raciborskii is a toxic bloom-forming cyanobacterium that occurs at tropical and temperate latitudes. Despite several reports from Africa, no data were previously available about its dynamics or toxic potential there. We therefore carried out a 1-year survey of the dynamics of C. raciborskii in the main water reservoir in Senegal, Lake Guiers. Cylindrospermopsis raciborskii never formed a bloom in this lake during the period studied, but was dominant during the dry season. The only observed bloom-forming species was a diatom, Fragilaria sp., which displayed a seasonal pattern contrary to that exhibited by C. raciborskii. Principal component analysis applied to environmental and phytoplankton data showed that high C. raciborskii biomasses were mainly related to high temperature and water column stability. Tests for C. raciborskii species-related toxicity and/or toxin synthesis were performed on 21 isolated clones. All the strains isolated tested negative in mouse toxicity bioassays, toxin analysis (MS/MS) and tests for known cylindrospermopsin genes (ps, pks). The limited number of isolates studied, and the occurrence of toxic and nontoxic clones in natural cyanobacterial populations, mean that we cannot conclude that there is no C. raciborskii-associated health risk in this drinking water reservoir.

First report in a river in France of the benthic cyanobacterium Phormidium favosum producing anatoxin-a associated with dog neurotoxicosis.

The first identification of anatoxin-a in a French lotic system is reported. Rapid deaths of dogs occurred in 2003 after the animals drank water from the shoreline of the La Loue River in eastern France. Sediments, stones and macrophytes surfaces at the margin of the river were covered by a thick biofilm containing large quantities of several benthic species of filamentous, non-heterocystous cyanobacteria. Known cyanotoxins, such as microcystins, saxitoxins and anatoxins were screened from biofilm samples by biochemical and analytical assays. A compound with similar UV spectra to the anatoxin-a standard was detected by high-performance liquid chromatography (HPLC) coupled with photo-diode array detector. This toxin was further identified by HPLC coupled with a UV detector and by electrospray ionisation-Quadrupole-Time-Of-Flight mass spectrometer, and confirmed by tandem mass spectrometry. These two techniques were necessary to discriminate anatoxin-a in phenylalanine-containing matrices such as liver samples of poisoned dogs. The toxin and the aromatic amino acid, phenylalanine, present the same pseudomolecular ion at m/z 166, but have differing fragmentation patterns, retention times and UV spectra. Finally, several cyanobacterial strains were isolated from the green biofilm and tested for anatoxin-a production. Phormidium favosum was identified as a new anatoxin-a producing species.