Neurometabolic profile of the amygdala in smokers assessed with 1H-magnetic resonance spectroscopy.

Tobacco smoking is one of the main causes of premature death worldwide and quitting success remains low, highlighting the need to understand the neurobiological mechanisms underlying relapse. Preclinical models have shown that the amygdala and glutamate play an important role in nicotine addiction. The aims of this study were to compare glutamate and other metabolites in the amygdala between smokers and controls, and between different smoking states. Furthermore, associations between amygdalar metabolite levels and smoking characteristics were explored. A novel non-water-suppressed proton magnetic resonance spectroscopy protocol was applied to quantify neurometabolites in 28 male smokers (≥15 cigarettes/day) and 21 non-smoking controls, matched in age, education, verbal IQ, and weekly alcohol consumption. Controls were measured once (baseline) and smokers were measured in a baseline state (1-3 h abstinence), during withdrawal (24 h abstinence) and in a satiation state (directly after smoking). Baseline spectroscopy data were compared between groups by independent t-tests or Mann-Whitney-U tests. Smoking state differences were investigated by repeated-measures analyses of variance (ANOVAs). Associations between spectroscopy data and smoking characteristics were explored using Spearman correlations. Good spectral quality, high anatomical specificity (98% mean gray matter) and reliable quantification of most metabolites of interest were achieved in the amygdala. Metabolite levels did not differ between groups, but smokers showed significantly higher glutamine levels at baseline than satiation. Glx levels were negatively associated with pack-years and smoking duration. In summary, this study provides first insights into the neurometabolic profile of the amygdala in smokers with high anatomical specificity. By applying proton magnetic resonance spectroscopy, neurometabolites in smokers during different smoking states and non-smoking controls were quantified reliably. A significant shift in glutamine levels between smoking states was detected, with lower concentrations in satiation than baseline. The negative association between Glx levels and smoking quantity and duration may imply altered glutamate homeostasis with more severe nicotine addiction.

In situ temperature determination using magnetic resonance spectroscopy thermometry for noninvasive postmortem examinations.

Magnetic resonance spectroscopy (MRS) thermometry offers a noninvasive, localized method for estimating temperature by leveraging the temperature-dependent chemical shift of water relative to a temperature-stable reference metabolite under suitable calibration. Consequentially, this technique has significant potential as a tool for postmortem MR examinations in forensic medicine and pathology. In these examinations, the deceased are examined at a wide range of body temperatures, and MRS thermometry may be used for the temperature adjustment of magnetic resonance imaging (MRI) protocols or for corrections in the analysis of MRI or MRS data. However, it is not yet clear to what extent postmortem changes may influence temperature estimation with MRS thermometry. In addition, N-acetylaspartate, which is commonly used as an in vivo reference metabolite, is known to decrease with increasing postmortem interval (PMI). This study shows that lactate, which is not only present in significant amounts postmortem but also has a temperature-stable chemical shift, can serve as a suitable reference metabolite for postmortem MRS thermometry. Using lactate, temperature estimation in postmortem brain tissue of severed sheep heads was accurate up to 60 h after death, with a mean absolute error of less than 0.5°C. For this purpose, published calibrations intended for in vivo measurements were used. Although postmortem decomposition resulted in severe metabolic changes, no consistent deviations were observed between measurements with an MR-compatible temperature probe and MRS thermometry with lactate as a reference metabolite. In addition, MRS thermometry was applied to 84 deceased who underwent a MR examination as part of the legal examination. MRS thermometry provided plausible results of brain temperature in comparison with rectal temperature. Even for deceased with a PMI well above 60 h, MRS thermometry still provided reliable readings. The results show a good suitability of MRS thermometry for postmortem examinations in forensic medicine.

Effects of Psychotherapy on Glutamatergic Neurotransmission.

INTRODUCTION: Psychodynamic psychotherapy is an effective and widely used treatment for major depressive disorder (MDD); however, little is known about neurobiological changes associated with induced symptom improvement. METHODS: Proton magnetic resonance spectroscopy with a two-dimensional J-resolved sequence served to test the relationship between glutamate (Glu) and glutamine (Gln) levels, measured separately in pregenual anterior cingulate cortex (pgACC) and the anterior midcingulate cortex (aMCC) as a control region, with change in depression symptoms after 6 months of weekly psychodynamic psychotherapy sessions in MDD patients. Depressed (N = 45) and healthy (N = 30) subjects participated in a baseline proton magnetic resonance spectroscopy measurement and a subgroup of MDD subjects (N = 21) then received once-a-week psychodynamic psychotherapy and participated in a second proton magnetic resonance spectroscopy measurement after 6 months. Change in depression symptoms was assessed using the Hamilton Depression Rating Scale (HAMD). RESULTS: Higher pretreatment pgACC Gln concentrations in MDD patients compared to healthy controls were associated with symptom severity. Patients and controls did not differ regarding Gln levels in aMCC nor regarding Glu levels in both regions. The association of pgACC Gln concentration and severity of depressive symptoms was reversed after 6 months of psychotherapy in MDD subjects. Regarding Gln in aMCC as well as Glu in both regions, there were no significant associations with improvement of depressive symptoms in the course of psychotherapy. DISCUSSION: Findings indicate specific regional effects of psychodynamic psychotherapy on glutamatergic neurotransmission and thereby highlight the key role of the pgACC in both depression pathophysiology and recovery.

Impaired glutamate homeostasis in the nucleus accumbens in human cocaine addiction.

Cocaine addiction is characterized by overwhelming craving for the substance, which drives its escalating use despite adverse consequences. Animal models suggest a disrupted glutamate homeostasis in the nucleus accumbens to underlie addiction-like behavior. After chronic administration of cocaine, rodents show decreased levels of accumbal glutamate, whereas drug-seeking reinstatement is associated with enhanced glutamatergic transmission. However, due to technical obstacles, the role of disturbed glutamate homeostasis for cocaine addiction in humans remains only partially understood, and accordingly, no approved pharmacotherapy exists. Here, we applied a tailored proton magnetic resonance spectroscopy protocol that allows glutamate quantification within the human nucleus accumbens. We found significantly reduced basal glutamate concentrations in the nucleus accumbens in cocaine-addicted (N = 26) compared with healthy individuals (N = 30), and increased glutamate levels during cue-induced craving in cocaine-addicted individuals compared with baseline. These glutamatergic alterations, however, could not be significantly modulated by a short-term challenge of N-acetylcysteine (2400 mg/day on 2 days). Taken together, our findings reveal a disturbed accumbal glutamate homeostasis as a key neurometabolic feature of cocaine addiction also in humans. Therefore, we suggest the glutamatergic system as a promising target for the development of novel pharmacotherapies, and in addition, as a potential biomarker for a personalized medicine approach in addiction.

Neurometabolic alterations in the nucleus accumbens of smokers assessed with 1 H magnetic resonance spectroscopy: The role of glutamate and neuroinflammation.

Tobacco use is one of the leading causes of premature death and morbidity worldwide. For smokers trying to quit, relapse rates are high, even after prolonged periods of abstinence. Recent findings in animal models highlight the role of alterations in glutamatergic projections from the prefrontal cortex onto the nucleus accumbens (NAc) in relapse vulnerability. Moreover, inflammatory responses in the NAc have been reported during withdrawal. A novel proton magnetic resonance spectroscopy (1 H-MRS) protocol was applied in humans to measure molar concentrations for glutamate, its sum with glutamine (Glx), and myoinositol plus glycine (mI + Gly) in the NAc (19 smokers, 20 matched controls). Smokers were measured at baseline and during withdrawal and satiation. No difference between groups or smoking states was found for glutamate or Glx, but, in smokers, stronger craving and more severe nicotine dependence were associated with lower baseline glutamate and Glx levels, respectively. Interestingly, mI + Gly concentrations were higher during withdrawal than baseline and correlated negatively with nicotine dependence severity and pack years of smoking. The lack of glutamatergic changes between groups and smoking states may imply that glutamate homeostasis is not significantly altered in smokers or that changes are too small for detection by 1 H-MRS. Moreover, the observed increase in mI + Gly may imply that neuroinflammatory processes occur in the NAc during nicotine withdrawal. These findings shed light on neurobiological relapse mechanisms in smokers and may provide the opportunity to develop more effective treatment options targeting the glutamate and neuroinflammation system.

Noninvasive in situ proton MRS in muscle tissue and bone marrow as a novel approach to identify previous freezing in a completely thawed cadaver.

The temporary or permanent storage of human bodies in freezers following a homicide is a documented method for criminal disposal of human corpses. In these cases, the detection of characteristics which indicate that a thawed cadaver or body part was previously frozen provides crucial information for forensic casework. Previous histological and radiological approaches to detect characteristics of previous freezing are based on the formation of bubble-like gas patterns, which are difficult to distinguish from common postmortem gas formation in the course of decomposition. The objective of this study was to detect changes in the muscle tissue and in the bone marrow after freezing and thawing by means of in situ proton magnetic resonance spectroscopy (1 H-MRS) to provide a noninvasive approach to detect postfreezing alterations in human cadavers. In this experimental study, the hind legs of seven sheep were used as substitutes for human tissue. One hind leg underwent 1 H-MRS before and daily after storage in a deep freezer (-20°C) and complete thawing at room temperature (study group: n = 7). The opposite hind leg was kept at room temperature and was measured daily (control group: n = 7). Spectra and relaxation times were measured using single voxel measurements in the muscle tissue and in the bone marrow. 1 H-MRS revealed several changes in the muscle tissue and in the bone marrow after freezing and thawing. A strongly reduced peak area ratio (<20) between bulk methylene and olefinic and glycerol methine and a reduced T2 relaxation time for bulk methylene (<45 ms) measured in the bone marrow were found to be indicators that a sheep leg was previously frozen and thawed independent of the postmortem interval. Noninvasive in situ 1 H-MRS in the bone marrow potentially provides a new method for detecting previous freezing or extreme cooling in cadavers.

Noninvasive analysis and identification of an intramuscular fluid collection by postmortem 1H-MRS in a case of a fatal motor vehicle accident.

In a case of a fatal traffic accident, a suspicious finding was identified in the muscular tissue of the left thigh by whole-body postmortem computed tomography. To better interpret the finding, the lower extremities were investigated by magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (1H-MRS). MRI revealed the presence of an evenly distributed intramuscular fluid and 1H-MRS of a volume within the fluid detected concentrations of acetate and lactate. The fluid was assumed to be an extravasation of an intraosseous infusion, erroneously administered to the intermediate vastus of the left thigh during resuscitation, which was later confirmed when access to resuscitation protocols was granted. Further ex situ 1H-MRS investigations of five different infusion fluids showed the possible discrimination of the fluids and further indicated the unknown fluid to be a Ringer's acetate solution. This paper presents the case-based application of postmortem intramuscular 1H-MRS and introduces the possibility of its use to differentiate exo- and endogenic fluids for forensic interpretation. Further research for this method regarding problems in forensic pathology is needed.

Comparison of the beta-hydroxybutyrate, glucose, and lactate concentrations derived from postmortem proton magnetic resonance spectroscopy and biochemical analysis for the diagnosis of fatal metabolic disorders.

PURPOSE: The detection and quantification of metabolites relevant for the diagnosis of fatal metabolic disorders by proton magnetic resonance spectroscopy (1H-MRS) was recently demonstrated. This prospective study aimed to compare the concentrations of beta-hydroxybutyrate (BHB), glucose (GLC), and lactate (LAC) derived from both biochemical analyses and 1H-MRS for the diagnosis of fatal metabolic disorders. METHODS: In total, 20 cases with suspected fatal metabolic disorders were included in the study. For the agreement based on thresholds, the concentrations of BHB and GLC in the vitreous humor (VH) from the right vitreous and in cerebrospinal fluid (CSF) from the right lateral ventricle were derived from 1H-MRS and biochemical analyses. The predefined thresholds for pathological elevations were 2.5 mmol/l for BHB and 10 mmol/l for GLC based on the literature. In addition, concentrations of the same metabolites in white matter (WM) tissue from the corona radiata of the right hemisphere were analyzed experimentally using both methods. To enable the biochemical analysis, a dialysate of WM tissue was produced. For all three regions, the LAC concentration was determined by both methods. RESULTS: The conclusive agreement based on thresholds was almost perfect between both methods with only one disagreement in a total of 70 comparisons due to the interference of a ferromagnetic dental brace. The differences in the concentrations between both methods showed high standard deviations. Confidence intervals of the bias not including 0 were found in CSF-GLC (- 3.1 mmol/l), WM-GLC (1.1 mmol/l), and WM-LAC (- 6.5 mmol/l). CONCLUSION: Despite a considerable total error attributable to both methods, MRS derives the same forensic conclusions as conventional biochemical analyses. An adaptation of the protocol to reduce the detected errors and more data are needed for the long-term validation of MRS for the diagnosis of fatal metabolic disorders. The production of WM dialysates cannot be recommended due to high glycolytic loss.

Feasibility, technique and accuracy of ultrasound-guided transurethral injections into the urinary sphincter of female cadavers: proof of concept.

BACKGROUND: The injection of muscle precursor cells (MPC) into the external urinary sphincter muscle (EUS) is a promising therapeutic option for regenerative treatment of stress urinary incontinence (SUI). The objective of the present project was to conduct a pre-clinical trial to investigate the feasibility and accuracy of ultrasound (US) guided, transurethral injections into the EUS of female cadavers. METHODS: This is a prospective, anatomical, interventional and radiological cadaveric laboratory investigation. Two urologists performed transurethral US-guided injections to deliver nano-iron particles into the EUS. The intervention was performed in three unfixed, fresh female cadavers. Each cadaver received MRI before and CT as well as MRI of the pelvis after the injections. RESULTS: The precision and accumulation of nano-iron particles in the EUS was compared using a rating scale to evaluate left versus right and anterior versus posterior distribution in axial and sagittal orientation with US, MRI and CT. The accuracy of our US-guided injections into the anterior target region yielded 4 points on the rating scale. Adequate precision and accumulation of particles in the left versus right EUS were also demonstrated (3 vs. 3.33 points, respectively). Signal intensity in MRI revealed a mean ratio of 0.33 before and after injection. CT scans showed no relevant artefacts impairing the assessment. CONCLUSION: US-guided, transurethral injection into the EUS is feasible and imaging reveals a precise accumulation in the target region. Our method provides an appropriate approach to deliver MPC in the EUS muscle for a regenerative treatment of SUI in the near future.

Noninvasive 7 tesla MRI of fatal craniocerebral gunshots - a glance into the future of radiologic wound ballistics.

Compared to computed tomography (CT), magnetic resonance imaging (MRI) provides superior visualization of the soft tissue. Recently, the first 7 Tesla (7 T) MRI scanner was approved for clinical use, which will facilitate access to these ultra-high-field MRI scanners for noninvasive examinations and scientific studies on decedents. 7 T MRI has the potential to provide a higher signal-to-noise ratio (SNR), a characteristic that can be directly exploited to improve image quality and invest in attempts to increase resolution. Therefore, evaluating the diagnostic potential of 7 T MRI for forensic purposes, such as assessments of fatal gunshot wounds, was deemed essential. In this article, we present radiologic findings obtained for craniocerebral gunshot wounds in three decedents. The decedents were submitted to MRI examinations using a 7 T MRI scanner that has been approved for clinical use and a clinical 3 T MRI scanner for comparison. We focused on detecting tiny injuries beyond the wound tract caused by temporary cavitation, such as microbleeds. Additionally, 7 T T2-weighted MRI highlighted a dark (hypo intense) zone beyond the permanent wound tract, which was attributed to increased amounts of paramagnetic blood components in damaged tissue. Microbleeds were also detected adjacent to the wound tract in the white matter on 7 T MRI. Based on the findings of radiologic assessments, the advantages and disadvantages of postmortem 7 T MRI compared to 3 T MRI are discussed with regard to investigations of craniocerebral gunshot wounds as well as the potential role of 7 T MRI in the future of forensic science.

Synergy of CT and MRI in detecting trajectories of lodged bullets in decedents and potential hazards concerning the heating and movement of bullets during MRI.

The purpose of this study was to assess the value of magnetic resonance imaging (MRI) in addition to computed tomography (CT) in gunshot wound cases with bullets or pellets lodged inside the head. In this context, the potential heating and movement of the lodged bullets were additionally investigated using animal models. Eleven forensic cases of penetrating gunshot wounds underwent CT and MRI. The data of each imaging modality were reviewed according to the following relevant characteristics: bony lesion at the entrance, intracranial bone fragments, intracranial metal fragments, gunshot residues, the wound channel and the severity of metal artifacts. Four-point Likert scales were used for the assessment. The heating of projectiles and their magnetic field interactions with the static magnetic field were assessed using animal models. MRI presented major advantages in cases with transversal trajectories and non-ferromagnetic bullets compared to CT. In general, MRI enabled a clear visualization of the wound channel and gunshot-related soft tissue injuries. An image fusion of CT and MRI datasets demonstrated the individual strengths of both modalities. Radio frequency (RF)-induced heating due to bullets lodged inside the brain tissue was invalidated. The likelihood of ferromagnetic projectile migration inside brain tissue is low. MRI of decedents with a bullet lodged inside their heads is viable and provides a valuable supplement to CT. The in situ, noninvasive depiction of the wound channel and gunshot-related soft tissue injuries on MRI can contribute to the knowledge of wound ballistics.

In situ postmortem ethanol quantification in the cerebrospinal fluid by non-water-suppressed proton MRS.

Determination of the ethanol concentration in corpses with MRS would allow a reproducible forensic assessment by which evidence is collected in a noninvasive manner. However, although MRS has been successfully used to detect ethanol in vivo, it has not been applied to postmortem ethanol quantification in situ. The present study examined the feasibility of the noninvasive measurement of the ethanol concentration in human corpses with MRS. A total of 15 corpses with suspected alcohol consumption before demise underwent examination in a 3 T whole body scanner. To address the partial overlap of the ethanol and lactate signal in the postmortem spectrum, non-water-suppressed single voxel spectra were recorded in the cerebrospinal fluid (CSF) of the left lateral ventricle via the metabolite cycling technique. The ethanol signals were quantified using the internal water as reference standard, as well as based on a reference signal acquired in a phantom. The measured values were compared with biochemically determined concentrations in the blood (BAC) and CSF (CSFAC). In 8 of the 15 corpses a BAC above zero was determined (range 0.03-1.68 g/kg). In all of these 8 corpses, ethanol was measured in CSF with the proposed MRS protocol. The two applied MRS calibration strategies resulted in similar concentrations. However, the MRS measurements generally overestimated the ethanol concentration by 0.09 g/kg (4%) to 0.72 g/kg (45%) as compared with the CSFAC value. The presented MRS protocol allows the measurement of ethanol in the CSF in human corpses and provides an estimation of the ethanol concentration prior to autopsy. Observed deviations from biochemically determined concentrations are mainly explained by the approximate correction of the relaxation attenuation of the ethanol signal.

Identification of Bullets Based on Their Metallic Components and X-Ray Attenuation Characteristics at Different Energy Levels on CT.

OBJECTIVE. This study aimed to identify bullets on the basis of their metallic components and to distinguish between ferromagnetic and nonferromagnetic bullets using CT. MATERIALS AND METHODS. Eight ferromagnetic, steel-jacketed lead bullets, four nonferromagnetic, non-steel-jacketed lead bullets, and four nonferromagnetic solid bullets composed of copper or copper and zinc alloys which we refer to here as "Cu(Zn) bullets," were scanned by CT at 80, 100, 120, and 140 kVp. Attenuation values (in Hounsfield units) were measured on an extended CT scale (ECTS) in the core and at the edge of the bullets and were used to calculate the dual-energy index (DEI). RESULTS. Although all nonferromagnetic bullets significantly differed from ferromagnetic bullets, the significant differences were solely attributed to the higher DEI of solid Cu(Zn) bullets compared with that of all-lead bullets. The lead bullets with ferromagnetic, steel-containing jackets did not differ from the lead bullets with nonferromagnetic, non-steel-containing jackets on the basis of DEIs obtained from core and edge measurements. Solid Cu(Zn) bullets could be clearly distinguished from lead bullets regardless of the metallic components of the jackets using DEI calculations from CT numbers on an ECTS. The DEIs based on the dual-energy pair 120 and 140 kVp appear to be the most appropriate for distinguishing between these two types of bullets. CONCLUSION. This study provides new scientific knowledge regarding metals and their characteristics at different tube voltage levels. The abilities of clinically approved dual-energy CT allow differentiation of bullets composed of low-atomic-number (Z) metals from bullets composed of high-Z metals via DEI calculations from CT numbers on an ECTS.

Quantitative magnetic resonance spectroscopy at 3T based on the principle of reciprocity.

Quantification of magnetic resonance spectroscopy signals using the phantom replacement method requires an adequate correction of differences between the acquisition of the reference signal in the phantom and the measurement in vivo. Applying the principle of reciprocity, sensitivity differences can be corrected at low field strength by measuring the RF transmitter gain needed to obtain a certain flip angle in the measured volume. However, at higher field strength the transmit sensitivity may vary from the reception sensitivity, which leads to wrongly estimated concentrations. To address this issue, a quantification approach based on the principle of reciprocity for use at 3T is proposed and validated thoroughly. In this approach, the RF transmitter gain is determined automatically using a volume-selective power optimization and complemented with information from relative reception sensitivity maps derived from contrast-minimized images to correct differences in transmission and reception sensitivity. In this way, a reliable measure of the local sensitivity was obtained. The proposed method is used to derive in vivo concentrations of brain metabolites and tissue water in two studies with different coil sets in a total of 40 healthy volunteers. Resulting molar concentrations are compared with results using internal water referencing (IWR) and Electric REference To access In vivo Concentrations (ERETIC). With the proposed method, changes in coil loading and regional sensitivity due to B1 inhomogeneities are successfully corrected, as demonstrated in phantom and in vivo measurements. For the tissue water content, coefficients of variation between 2% and 3.5% were obtained (0.6-1.4% in a single subject). The coefficients of variation of the three major metabolites ranged from 3.4-14.5%. In general, the derived concentrations agree well with values estimated with IWR. Hence, the presented method is a valuable alternative for IWR, without the need for additional hardware such as ERETIC and with potential advantages in diseased tissue.

Postmortem 1H-MRS-Detection of Ketone Bodies and Glucose in Diabetic Ketoacidosis.

Diabetic ketoacidosis (DKA) is a metabolic complication of diabetes mellitus that takes a lethal course if untreated. In this way relevant to forensic medicine, secure diagnosis of DKA usually involves the evidence of elevated levels of glucose and the ketone bodies acetone, acetoacetate, and ß-hydroxybutyrate in corpse fluids. We conducted a postmortem hydrogen proton magnetic resonance spectroscopy (1H-MRS) in a case of lethal DKA. Distinctive resonances of all three ketone bodies as well as glucose were visible in spectra of cerebrospinal fluid, vitreous humor, and white matter. Estimated concentrations of ketone bodies and glucose supported the findings both of autopsy and biochemical analysis. Advantages of human postmortem 1H-MRS are the lack of movement and flow artifacts as well as lesser limitations of scan duration. Postmortem 1H-MRS is able to non-invasively measure concentrations of glucose and ketone bodies in small volumes of various regions of the brain. It may thus become a diagnostic tool for forensic investigations by quick determination of pathological metabolite concentrations in addition to conventional autopsy.

Elucidation of the downfield spectrum of human brain at 7 T using multiple inversion recovery delays and echo times.

PURPOSE: To characterize the downfield spectrum at 5-10 ppm in the human brain at a high magnetic field of 7 T. Knowledge of relaxation parameters is of interest for spectroscopy as well as chemical exchange-dependent saturation transfer experiments. METHODS: Water-suppressed spectra were recorded as echo time and inversion time series in healthy volunteers to investigate T2 and T1 values of downfield peaks in gray matter at 7T. The spectra were fitted in a two-dimensional fashion to a heuristic model of a series of Voigt lines, and the relaxation times were obtained for 12 peaks of interest. RESULTS: The mean T2 values averaged over the volunteers ranged from 24 to 158 ms, whereas the mean T1 values ranged from 0.22 to 2.40 s. Spectra of specific inversion and echo times revealed superposition of the amide peaks of N-acetylaspartate with short T2 and an inhomogeneously broadened component with longer T2 . CONCLUSIONS: T2 values were shorter than expected for most peaks, whereas T1 values had a very wide range; shorter relaxation times for some peaks suggests the presence of macromolecules. Most of the larger peaks seemed to be composed of overlapping components, because the Gaussian widths in the Voigt line shape descriptions were larger than expected based on field inhomogeneities. Magn Reson Med 78:11-19, 2016. © 2017 International Society for Magnetic Resonance in Medicine.

Accurate determination of brain metabolite concentrations using ERETIC as external reference.

Magnetic Resonance Spectroscopy (MRS) can provide in vivo metabolite concentrations in standard concentration units if a reliable reference signal is available. For 1 H MRS in the human brain, typically the signal from the tissue water is used as the (internal) reference signal. However, a concentration determination based on the tissue water signal most often requires a reliable estimate of the water concentration present in the investigated tissue. Especially in clinically interesting cases, this estimation might be difficult. To avoid assumptions about the water in the investigated tissue, the Electric REference To access In vivo Concentrations (ERETIC) method has been proposed. In this approach, the metabolite signal is compared with a reference signal acquired in a phantom and potential coil-loading differences are corrected using a synthetic reference signal. The aim of this study, conducted with a transceiver quadrature head coil, was to increase the accuracy of the ERETIC method by correcting the influence of spatial B1 inhomogeneities and to simplify the quantification with ERETIC by incorporating an automatic phase correction for the ERETIC signal. Transmit field ( B1+) differences are minimized with a volume-selective power optimization, whereas reception sensitivity changes are corrected using contrast-minimized images of the brain and by adapting the voxel location in the phantom measurement closely to the position measured in vivo. By applying the proposed B1 correction scheme, the mean metabolite concentrations determined with ERETIC in 21 healthy subjects at three different positions agree with concentrations derived with the tissue water signal as reference. In addition, brain water concentrations determined with ERETIC were in agreement with estimations derived using tissue segmentation and literature values for relative water densities. Based on the results, the ERETIC method presented here is a valid tool to derive in vivo metabolite concentration, with potential advantages compared with internal water referencing in diseased tissue.

Neurochemical profile of the human cervical spinal cord determined by MRS.

MRS enables insight into the chemical composition of central nervous system tissue. However, technical challenges degrade the data quality when applied to the human spinal cord. Therefore, to date detection of only the most prominent metabolite resonances has been reported in the healthy human spinal cord. The aim of this investigation is to provide an extended metabolic profile including neurotransmitters and antioxidants in addition to metabolites involved in the energy and membrane metabolism of the human cervical spinal cord in vivo. To achieve this, data quality was improved by using a custom-made, cervical detector array together with constructive averaging of a high number of echo signals, which is enabled by the metabolite cycling technique at 3T. In addition, the improved spinal cord spectra were extensively cross-validated, in vivo, post-mortem in situ and ex vivo. Reliable identification of up to nine metabolites was achieved in group analyses for the first time. Distinct features of the spinal cord neurochemical profile, in comparison with the brain neurotransmission system, include decreased concentrations of the sum of glutamate and glutamate and increased concentrations of aspartate, γ-amino-butyric acid, scyllo-inositol and the sum of myo-inositol and glycine.

Ballistic study on the penetration potential and injury potential of different bullet types in the use of a newly developed bullet shooting stunner for adequate stunning of heavy cattle.

Introduction: Recently, a special bullet shooting stunner for heavy cattle has been developed that fires a bullet instead of a bolt. In the search for a suitable ammunition, the following criteria must be met: First, the energy of the bullet must be sufficient to penetrate the thick frontal bones of heavy cattle. Second, the injury potential at the corresponding penetration depth should preferably be large in order to damage brain tissue relevant to stunning. Third, the bullet must not perforate the occipital bone (over-penetration). Methods: Four different bullet types [Hornady FTX, Hydra-Shok, Black Mamba, and a common full metal jacket (FMJ) bullet] were evaluated in a series of experiments on soap blocks and removed bone plates followed by computed tomography examinations. Penetration potential was evaluated in terms of kinetic energy relative to the caliber of the bullet, i.e., mean energy density (ED). Injury potential was evaluated by the mean extent of the cavity volume (e CV ) at the relevant penetration depth of 5.5 to 7.5 cm in the soap block. Results: All four bullet types passed through the frontal bone plate. The ED was 17.50 J/mm2 (Hornady FTX), 17.46 J/mm2 (Hydra-Shok), 13.47 J/mm2 (Black Mamba), and 13.47 J/mm2 (FMJ). The Hornady FTX and the Hydra-Shok each fragmented heavily. The FMJ was excluded after three experiments due to over-penetrations. The e CV was e CV = 3.77 cm2 (Hornady FTX), 2.71 cm2 (Hydra-Shok), and 1.31 cm2 (Black Mamba), with a significant difference (p = 0.006) between the Hornady FTX and the Black Mamba. Discussion: For use in heavy cattle, the Hornady FTX and the Hydra-Shok are recommended due to the larger e CV than the Black Mamba.

Accumbal-thalamic connectivity and associated glutamate alterations in human cocaine craving: A state-dependent rs-fMRI and 1H-MRS study.

Craving is a core symptom of cocaine use disorder and a major factor for relapse risk. To date, there is no pharmacological therapy to treat this disease or at least to alleviate cocaine craving as a core symptom. In animal models, impaired prefrontal-striatal signalling leading to altered glutamate release in the nucleus accumbens appear to be the prerequisite for cocaine-seeking. Thus, those network and metabolic changes may constitute the underlying mechanisms for cocaine craving and provide a potential treatment target. In humans, there is recent evidence for corresponding glutamatergic alterations in the nucleus accumbens, however, the underlying network disturbances that lead to this glutamate imbalance remain unknown. In this state-dependent randomized, placebo-controlled, double-blinded, cross-over multimodal study, resting state functional magnetic resonance imaging in combination with small-voxel proton magnetic resonance spectroscopy (voxel size: 9.4 × 18.8 × 8.4 mm3) was applied to assess network-level and associated neurometabolic changes during a non-craving and a craving state, induced by a custom-made cocaine-cue film, in 18 individuals with cocaine use disorder and 23 healthy individuals. Additionally, we assessed the potential impact of a short-term challenge of N-acetylcysteine, known to normalize disturbed glutamate homeostasis and to thereby reduce cocaine-seeking in animal models of addiction, compared to a placebo. We found increased functional connectivity between the nucleus accumbens and the dorsolateral prefrontal cortex during the cue-induced craving state. However, those changes were not linked to alterations in accumbal glutamate levels. Whereas we additionally found increased functional connectivity between the nucleus accumbens and a midline part of the thalamus during the cue-induced craving state. Furthermore, obsessive thinking about cocaine and the actual intensity of cocaine use were predictive of cue-induced functional connectivity changes between the nucleus accumbens and the thalamus. Finally, the increase in accumbal-thalamic connectivity was also coupled with craving-related glutamate rise in the nucleus accumbens. Yet, N-acetylcysteine had no impact on craving-related changes in functional connectivity. Together, these results suggest that connectivity changes within the fronto-accumbal-thalamic loop, in conjunction with impaired glutamatergic transmission, underlie cocaine craving and related clinical symptoms, pinpointing the thalamus as a crucial hub for cocaine craving in humans.