Hepatic passaging of NRAS-mutant melanoma influences adhesive properties and metastatic pattern.

BACKGROUND: Liver metastasis is a poor prognostic factor for treatment of advanced cutaneous melanoma with either immunotherapy or targeted therapies. In this study we focused on NRAS mutated melanoma, a cohort with high unmet clinical need. METHODS: WT31 melanoma was repeatedly passaged over the liver after intravenous injections five times generating the subline WT31_P5IV. The colonization of target organs, morphology, vascularization and the gene expression profiles of metastases were analyzed. RESULTS: After intravenous injection lung metastasis was significantly decreased and a trend towards increased liver metastasis was detected for WT31_P5IV as compared to parental WT31. Besides, the ratio of lung to liver metastases was significantly smaller. Histology of lung metastases revealed reduced proliferation of WT31_P5IV in relation to WT31 while both size and necrotic areas were unaltered. Liver metastases of both sublines showed no differences in vascularization, proliferation or necrosis. To identify tumor-intrinsic factors that altered the metastatic pattern of WT31_P5IV RNA sequencing was performed and revealed a differential regulation of pathways involved in cell adhesion. Ex vivo fluorescence imaging confirmed that initial tumor cell retention in the lungs was significantly reduced in WT31_P5IV in comparison to WT31. CONCLUSION: This study demonstrates that tumor-intrinsic properties influencing the metastatic pattern of NRAS mutated melanoma are strongly affected by hepatic passaging and the hematogenous route tumor cells take. It has implications for the clinical setting as such effects might also occur during metastatic spread or disease progression in melanoma patients.

Perfusion and T2 Relaxation Time as Predictors of Severity and Outcome in Sepsis-Associated Acute Kidney Injury: A Preclinical MRI Study.

BACKGROUND: Preventing sepsis-associated acute kidney injury (S-AKI) can be challenging because it develops rapidly and is often asymptomatic. Probability assessment of disease progression for therapeutic follow-up and outcome are important to intervene and prevent further damage. PURPOSE: To establish a noninvasive multiparametric MRI (mpMRI) tool, including T1 , T2 , and perfusion mapping, for probability assessment of the outcome of S-AKI. STUDY TYPE: Preclinical randomized prospective study. ANIMAL MODEL: One hundred and forty adult female SD rats (65 control and 75 sepsis). FIELD STRENGTH/SEQUENCE: 9.4T; T1 and perfusion map (FAIR-EPI) and T2 map (multiecho RARE). ASSESSMENT: Experiment 1: To identify renal injury in relation to sepsis severity, serum creatinine levels were determined (31 control and 35 sepsis). Experiment 2: Animals underwent mpMRI (T1 , T2 , perfusion) 18 hours postsepsis. A subgroup of animals was immediately sacrificed for histology examination (nine control and seven sepsis). Result of mpMRI in follow-up subgroup (25 control and 33 sepsis) was used to predict survival outcomes at 96 hours. STATISTICAL TESTS: Mann-Whitney U test, Spearman/Pearson correlation (r), P < 0.05 was considered statistically significant. RESULTS: Severely ill septic animals exhibited significantly increased serum creatinine levels compared to controls (70 ± 30 vs. 34 ± 9 µmol/L, P < 0.0001). Cortical perfusion (480 ± 80 vs. 330 ± 140 mL/100 g tissue/min, P < 0.005), and cortical and medullary T2 relaxation time constants were significantly reduced compared to controls (41 ± 4 vs. 37 ± 5 msec in cortex, P < 0.05, 52 ± 7 vs. 45 ± 6 msec in medulla, P < 0.05). The combination of cortical T2 relaxation time constants and perfusion results at 18 hours could predict survival outcomes at 96 hours with high sensitivity (80%) and specificity (73%) (area under curve of ROC = 0.8, Jmax = 0.52). DATA CONCLUSION: This preclinical study suggests combined T2 relaxation time and perfusion mapping as first line diagnostic tool for treatment planning. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY STAGE: 2.

Investigation of Experimental Endovascular Air Embolisms Using a New Model for the Generation and Detection of Highly Calibrated Micro Air Bubbles.

BACKGROUND: Air embolism (AE), especially when affecting the brain, is an underrated and potentially life-threatening complication in various endovascular interventions. This study aims to investigate experimental AEs using a new model to generate micro air bubbles (MAB), to assess the impact of a catheter on these MAB, and to demonstrate the applicability of this model in vivo. MATERIALS AND METHODS: Micro air bubbles were created using a system based on microfluidic channels. The MAB were detected and analyzed automatically. Micro air bubbles, with a target size of 85 µm, were generated and injected through a microcatheter. The MAB diameters proximal and distal to the catheter were assessed and compared. In a subsequent in vivo application, 2000 MAB were injected into the aorta (at the aortic valve) and into the common carotid artery (CCA) of a rat, respectively, using a microcatheter, resembling AE occurring during cardiovascular interventions. RESULTS: Micro air bubbles with a highly calibrated size could be successfully generated (median: 85.5 µm, SD 1.9 µm). After passage of the microcatheter, the MAB were similar in diameter (median: 86.6 µm) but at a lower number (60.1% of the injected MAB) and a substantially higher scattering of diameters (SD 29.6 µm). In vivo injection of MAB into the aorta resulted in cerebral microinfarctions in both hemispheres, whereas injection into the CCA caused exclusively ipsilateral microinfarctions. CONCLUSION: Using this new AE model, MAB can be generated precisely and reproducibly, resulting in cerebral microinfarctions. This model is feasible for further studies on the pathophysiology and prevention of AE in cardiovascular procedures.

Whole-body diffusion magnetic resonance imaging with simultaneous multi-slice excitation in children and adolescents.

BACKGROUND: Whole-body magnetic resonance imaging (WB-MRI) is an increasingly used guideline-based imaging modality for oncological and non-oncological pathologies during childhood and adolescence. While diffusion-weighted imaging (DWI), a part of WB-MRI, enhances image interpretation and improves sensitivity, it also requires the longest acquisition time during a typical WB-MRI scan protocol. Interleaved short tau inversion recovery (STIR) DWI with simultaneous multi-slice (SMS) acquisition is an effective way to speed up examinations. OBJECTIVE: In this study of children and adolescents, we compared the acquisition time, image quality, signal-to-noise ratio (SNR) and apparent diffusion coefficient (ADC) values of an interleaved STIR SMS-DWI sequence with a standard non-accelerated DWI sequence for WB-MRI. MATERIALS AND METHODS: Twenty children and adolescents (mean age: 13.9 years) who received two WB-MRI scans at a maximum interval of 18 months, consisting of either standard DWI or SMS-DWI MRI, respectively, were included. For quantitative evaluation, the signal-to-noise ratio (SNR) was determined for b800 images and ADC maps of seven anatomical regions. Image quality evaluation was independently performed by two experienced paediatric radiologists using a 5-point Likert scale. The measurement time per slice stack, pause between measurements including shim and total measurement time of DWI for standard DWI and SMS-DWI were extracted directly from the scan data. RESULTS: When including the shim duration, the acquisition time for SMS-DWI was 43% faster than for standard DWI. Qualitatively, the scores of SMS-DWI were higher in six locations in the b800 images and four locations in the ADC maps. There was substantial agreement between both readers, with a Cohen's kappa of 0.75. Quantitatively, the SNR in the b800 images and the ADC maps did not differ significantly from one another. CONCLUSION: Whole body-MRI with SMS-DWI provided equivalent image quality and reduced the acquisition time almost by half compared to the standard WB-DWI protocol.

[Is JAK inhibition an option in the treatment of interstitial lung disease in rheumatoid arthritis?] / Ist JAK-Hemmung eine Option in der Behandlung der interstitiellen Lungenerkrankung bei einer rheumatoiden Arthritis?

A 69-year-old male patient with seropositive erosive rheumatoid arthritis (RA) presented to our clinic due to progressive dyspnea. High-resolution computed tomography (HRCT) and immunological bronchioalveolar lavage revealed ground-glass opacities and a lymphocytic alveolitis caused by interstitial lung disease (ILD) in RA. Considering previous forms of treatment, disease-modifying antirheumatic drug (DMARD) treatment was switched to tofacitinib. Tofacitinib treatment demonstrated a 33% reduction in ground-glass opacities by artificial intelligence-based quantification of pulmonary HRCT over the course of 6 months, which was associated with an improvement in dyspnea symptoms. In conclusion, tofacitinib represents an effective anti-inflammatory therapeutic option in the treatment of RA-ILD.

CaM kinase II regulates cardiac hemoglobin expression through histone phosphorylation upon sympathetic activation.

Sympathetic activation of ß-adrenoreceptors (ß-AR) represents a hallmark in the development of heart failure (HF). However, little is known about the underlying mechanisms of gene regulation. In human ventricular myocardium from patients with end-stage HF, we found high levels of phosphorylated histone 3 at serine-28 (H3S28p). H3S28p was increased by inhibition of the catecholamine-sensitive protein phosphatase 1 and decreased by ß-blocker pretreatment. By a series of in vitro and in vivo experiments, we show that the ß-AR downstream protein kinase CaM kinase II (CaMKII) directly binds and phosphorylates H3S28. Whereas, in CaMKII-deficient myocytes, acute catecholaminergic stimulation resulted in some degree of H3S28p, sustained catecholaminergic stimulation almost entirely failed to induce H3S28p. Genome-wide analysis of CaMKII-mediated H3S28p in response to chronic ß-AR stress by chromatin immunoprecipitation followed by massive genomic sequencing led to the identification of CaMKII-dependent H3S28p target genes. Forty percent of differentially H3S28p-enriched genomic regions were associated with differential, mostly increased expression of the nearest genes, pointing to CaMKII-dependent H3S28p as an activating histone mark. Remarkably, the adult hemoglobin genes showed an H3S28p enrichment close to their transcriptional start or end sites, which was associated with increased messenger RNA and protein expression. In summary, we demonstrate that chronic ß-AR activation leads to CaMKII-mediated H3S28p in cardiomyocytes. Thus, H3S28p-dependent changes may play an unexpected role for cardiac hemoglobin regulation in the context of sympathetic activation. These data also imply that CaMKII may be a yet unrecognized stress-responsive regulator of hematopoesis.

A Novel Dipotassium Hydrogen Phosphate Phantom for Calibrating Computed Tomographic Bone Density Measurements in Birds.

The objective of this study was to construct a calibration phantom for bone mineral density (BMD) measurements adapted to avian anatomy by quantitative computed tomography. The determination of BMD is important to assess avian osteoporosis in poultry at production facilities and to study biological features in association with flight patterns in birds. Quantitative computed tomography measured in Hounsfield units is a well-established technique for BMD measurements. Translation of Hounsfield units into the International System of Units (mg/cm3) requires the use of a calibration phantom. Although calibration phantoms for routine use in humans are commercially available, phantoms suited to avian anatomy are not. A liquid dipotassium hydrogen phosphate calibration standard was constructed out of commercially available materials, easily allowing for variations in size, bone diameter, and adaptation to avian skeletal anatomy. Periodically, quantitative computed tomography scans were performed to monitor constant correlation to the calibration standard over 3 months and to monitor for the potential influence of gas bubbling and water evaporation in the rods on BMD measurements. Finally, the calibration phantom was tested for BMD measurements with carcasses from 2 bird species, including 3 peregrine falcons (Falco peregrinus; 2 juvenile males, 1 adult female with inactive reproductive status) and 4 Eurasian kestrels (Falco tinnunculus; 1 juvenile and 2 adult males, 1 adult female with inactive reproductive status). Results demonstrated stability of the calibration phantom without the need to refill or replace rods, plus a stable correlation line (R 2 = .99) over the 3-month evaluation period. It was possible to place the phantom directly on the bird carcasses, close to the measured bones, to improve BMD analysis. As evaluated, the phantom appeared to be adaptive to avian skeletal anatomy. Moreover, it was possible to build the phantom within 24 hours from commercially available materials.

Biomechanical comparison of screw and suture fixation for temporary transarticular tarsocrural immobilization in canine cadavers.

OBJECTIVE: To biomechanically evaluate an ultrahigh-molecular-weight polyethylene (UWMWPE) suture for temporary tarsocrural immobilization as a potential alternative to the existing surgical method, which uses a 4.5 AO/ASIF calcaneotibial cortical screw. STUDY DESIGN: Randomized in vitro biomechanical study. ANIMALS: Pelvic limbs (n = 20) from 10 euthanized adult dogs (average bodyweight of 29.9 kg ± 1.9 kg). METHODS: Tarsocrural joints were immobilized either with a 4.5 AO/ASIF screw or a 1 mm diameter UHMWPE suture. Limbs were loaded with 60 N, 120 N, and 180 N, each for 100 cycles. After cyclic loading, immobilizations were loaded until failure. Load at failure and mode of failure were recorded. RESULTS: Both immobilization methods remained intact during loading with 60 N, 120 N, and 180 N. Mean and standard deviation (± SD) failure loads were higher for the cortical screw (524.9 N ± 148.7 N) than for the UHMWPE suture (387.8 N ± 105.6 N), P = .0084. The stiffness of both systems was equivalent. CONCLUSION: Both techniques were suitable for transarticular tarsocrural immobilization in large-breed canine cadavers. While load at failure was higher for the screw compared to the suture construct, stiffness of both methods was comparable based on the laboratory settings in this in vitro study. CLINICAL SIGNIFICANCE: Based on the results of this study, both the cortical screw and the UHMWPE suture stabilized the tibiotarsal joint at clinically relevant loads. However, when exposed to high loads, the cortical screw provides increased tibiotarsal stability. Comparative data need to be collected prior to widespread use of UHMWPE sutures in clinical cases.

Immersion of Achilles tendon in phosphate-buffered saline influences T1 and T2 * relaxation times: An ex vivo study.

Robust mapping of relaxation parameters in ex vivo tissues is based on hydration and therefore requires control of the tissue treatment to ensure tissue integrity and consistent measurement conditions over long periods of time. One way to maintain the hydration of ex vivo tendon tissue is to immerse the samples in a buffer solution. To this end, various buffer solutions have been proposed; however, many appear to influence the tissue relaxation times, especially with prolonged exposure. In this work, ovine Achilles tendon tissue was used as a model to investigate the effect of immersion in phosphate-buffered saline (PBS) and the effects on the T1 and T2* relaxation times. Ex vivo samples were measured at 0 (baseline), 30 and 67 hours after immersion in PBS. Ultrashort echo time (UTE) imaging was performed using variable flip angle and echo train-shifted multi-echo imaging for T1 and T2* estimation, respectively. Compared with baseline, both T1 and T2* relaxation time constants increased significantly after 30 hours of immersion. T2* continued to show a significant increase between 30 and 67 hours. Both T1 and T2* tended to approach saturation at 67 hours. These results exemplify the relevance of stringently controlled tissue preparation and preservation techniques, both before and during MRI experiments.

Electrical Stimulation of the Anterior Thalamus for Epilepsy: Clinical Outcome and Analysis of Efficient Target.

OBJECTIVE: Deep brain stimulation (DBS) of the anterior thalamic complex (ANT) is an adjunctive therapy for pharmacoresistant epilepsy. To define the most efficient target in DBS for epilepsy, we investigate clinical data, position of leads, usability of atlas data compared to electric field modeling based on programming parameters. METHODS: Data from ten consecutive patients who underwent ANT-DBS were analyzed. The mammillothalamic tract (MTT), an internal landmark for direct stereotactic targeting, was segmented from MRI. Centers of stimulation were determined and their positions relative to ventricles and the MTT were analyzed. Two 3D thalamus atlases were transformed to segmented patient's thalami and proportions of activated nuclei were calculated. RESULTS: Our data indicate higher response rates with a center of stimulation 5 mm lateral to the wall of the third ventricle (R2 for reduction of focal seizure frequency and distance to the wall of the third ventricle = 0.48, p = 0.026). For reduction of focal seizures, a strong positive correlation with the dorsal distance to the midcommissural plane was found (R2 = 0.66, p = 0.004). In one 3D atlas, stimulation of internal medullary lamina (IML) correlated strongly positive with response rates, which, however, did not reach statistical significance (R2 = 0.69, p = 0.17 for tonic-clonic seizures). All electrical fields covered the diameter of the MTT. The position of the MTT in the thalamus was highly variable (range: x-coordinate 4.0 to 7.3 mm, y-coordinate -1.3 to 5.1 mm in AC-PC space). CONCLUSIONS: The distance of the active contact to the lateral wall of the third ventricle, MTT and the ventrodorsal distance to midcommissural plane appear to be relevant for optimal target planning. For reduction of focal seizure frequency, we found best response rates with a center of stimulation 5 mm lateral to the wall of the third ventricle, and a lead tip 10 mm dorsal of the midcommissural plane.

Predominance of hypoechoic tissue changes in nine dogs with malignant prostatic lymphoma.

Neoplasia of the prostate is relatively uncommon in dogs with adenocarcinoma being the most common type. Non-epithelial tumors are rare and only individual cases of malignant lymphoma affecting the prostate have been reported. The purpose of this multi-institutional, retrospective, descriptive study was to characterize the ultrasonographic features of canine prostatic lymphoma. Inclusion criteria were an abdominal ultrasound and cytological/histological diagnosis of malignant prostatic lymphoma. Ultrasonographic features were recorded based on the original ultrasonographic reports and consensus opinion of two readers on the available image sets retrospectively. Nine dogs met the inclusion criteria with a mean age of 6.5 years. Seven dogs were intact and two neutered. Subjective prostatomegaly was noted in all patients however not reproducible by objective measurements. Altered shape with rounded/irregular margins was detected in 78% of the cases. All prostates presented either diffuse (three dogs) or focal/periurethral (four dogs) and/or multifocal areas of hypoechogenicity (three dogs). In one dog, focal and multifocal hypoechoic changes co-occurred. Prostatic mineralization was not present in any of the cases. Ultrasonographic features of infiltrative disease of multiple organs and/or lymphadenopathy was found in all cases. Even though malignant lymphoma is rare in the prostate, it should be included in the list of differentials in patients with hypoechoic lesions/areas, altered shape, lack of mineralization of the prostatic parenchyma and evidence of multiorgan involvement.

Comparison of metabolic adaptations between endurance- and sprint-trained athletes after an exhaustive exercise in two different calf muscles using a multi-slice 31 P-MR spectroscopic sequence.

Measurements of exercise-induced metabolic changes, such as oxygen consumption, carbon dioxide exhalation or lactate concentration, are important indicators for assessing the current performance level of athletes in training science. With exercise-limiting metabolic processes occurring in loaded muscles, 31 P-MRS represents a particularly powerful modality to identify and analyze corresponding training-induced alterations. Against this background, the current study aimed to analyze metabolic adaptations after an exhaustive exercise in two calf muscles (m. soleus - SOL - and m. gastrocnemius medialis - GM) of sprinters and endurance athletes by using localized dynamic 31 P-MRS. In addition, the respiratory parameters VO2 and VCO2 , as well as blood lactate concentrations, were monitored simultaneously to assess the effects of local metabolic adjustments in the loaded muscles on global physiological parameters. Besides noting obvious differences between the SOL and the GM muscles, we were also able to identify distinct physiological strategies in dealing with the exhaustive exercise by recruiting two athlete groups with opposing metabolic profiles. Endurance athletes tended to use the aerobic pathway in the metabolism of glucose, whereas sprinters produced a significantly higher peak concentration of lactate. These global findings go along with locally measured differences, especially in the main performer GM, with sprinters revealing a higher degree of acidification at the end of exercise (pH 6.29 ± 0.20 vs. 6.57 ± 0.21). Endurance athletes were able to partially recover their PCr stores during the exhaustive exercise and seemed to distribute their metabolic activity more consistently over both investigated muscles. In contrast, sprinters mainly stressed Type II muscle fibers, which corresponds more to their training orientation preferring the glycolytic energy supply pathway. In conclusion, we were able to analyze the relation between specific local metabolic processes in loaded muscles and typical global adaptation parameters, conventionally used to monitor the training status of athletes, in two cohorts with different sports orientations.

DHODH modulates transcriptional elongation in the neural crest and melanoma.

Melanoma is a tumour of transformed melanocytes, which are originally derived from the embryonic neural crest. It is unknown to what extent the programs that regulate neural crest development interact with mutations in the BRAF oncogene, which is the most commonly mutated gene in human melanoma. We have used zebrafish embryos to identify the initiating transcriptional events that occur on activation of human BRAF(V600E) (which encodes an amino acid substitution mutant of BRAF) in the neural crest lineage. Zebrafish embryos that are transgenic for mitfa:BRAF(V600E) and lack p53 (also known as tp53) have a gene signature that is enriched for markers of multipotent neural crest cells, and neural crest progenitors from these embryos fail to terminally differentiate. To determine whether these early transcriptional events are important for melanoma pathogenesis, we performed a chemical genetic screen to identify small-molecule suppressors of the neural crest lineage, which were then tested for their effects on melanoma. One class of compound, inhibitors of dihydroorotate dehydrogenase (DHODH), for example leflunomide, led to an almost complete abrogation of neural crest development in zebrafish and to a reduction in the self-renewal of mammalian neural crest stem cells. Leflunomide exerts these effects by inhibiting the transcriptional elongation of genes that are required for neural crest development and melanoma growth. When used alone or in combination with a specific inhibitor of the BRAF(V600E) oncogene, DHODH inhibition led to a marked decrease in melanoma growth both in vitro and in mouse xenograft studies. Taken together, these studies highlight developmental pathways in neural crest cells that have a direct bearing on melanoma formation.

Behaviour of adipose-derived canine mesenchymal stem cells after superparamagnetic iron oxide nanoparticles labelling for magnetic resonance imaging.

BACKGROUND: Therapy with mesenchymal stem cells (MSCs) has been reported to provide beneficial effects in the treatment of neurological and orthopaedic disorders in dogs. The exact mechanism of action is poorly understood. Magnetic resonance imaging (MRI) gives the opportunity to observe MSCs after clinical administration. To visualise MSCs with the help of MRI, labelling with an MRI contrast agent is necessary. However, it must be clarified whether there is any negative influence on cell function and viability after labelling prior to clinical administration. RESULTS: For the purpose of the study, seven samples with canine adipose-derived stem cells were incubated with superparamagnetic iron oxide nanoparticles (SPIO: 319.2 µg/mL Fe) for 24 h. The internalisation of the iron particles occurred via endocytosis. SPIO particles were localized as free clusters in the cytoplasm or within lysosomes depending on the time of investigation. The efficiency of the labelling was investigated using Prussian blue staining and MACS assay. After 3 weeks the percentage of SPIO labelled canine stem cells decreased. Phalloidin staining showed no negative effect on the cytoskeleton. Labelled cells underwent osteogenic and adipogenic differentiation. Chondrogenic differentiation occurred to a lesser extent compared with a control sample. MTT-Test and wound healing assay showed no influence of labelling on the proliferation. The duration of SPIO labelling was assessed using a 1 Tesla clinical MRI scanner and T2 weighted turbo spin echo and T2 weighted gradient echo MRI sequences 1, 2 and 3 weeks after labelling. The hypointensity caused by SPIO lasted for 3 weeks in both sequences. CONCLUSIONS: An Endorem labelling concentration of 319.2 µg/mL Fe (448 µg/mL SPIO) had no adverse effects on the viability of canine ASCs. Therefore, this contrast agent could be used as a model for iron oxide labelling agents. However, the tracking ability in vivo has to be evaluated in further studies.

Self-gated cardiac Cine MRI of the rat on a clinical 3 T MRI system.

The ability to perform small animal functional cardiac imaging on clinical MRI scanners may be of particular value in cases in which the availability of a dedicated high field animal MRI scanner is limited. Here, we propose radial MR cardiac imaging in the rat on a whole-body clinical 3 T scanner in combination with interspersed projection navigators for self-gating without any additional external triggering requirements for electrocardiogram (ECG) and respiration. Single navigator readouts were interspersed using the same TR and a high navigator frequency of 54 Hz into a radial golden-angle acquisition. The extracted navigator function was thresholded to exclude data for reconstruction from inhalation phases during the breathing cycle, enabling free breathing acquisition. To minimize flow artifacts in the dynamic cine images a center-out half echo radial acquisition scheme with ramp sampling was used. Navigator functions were derived from the corresponding projection navigator data from which both respiration and cardiac cycles were extracted. Self-gated cine acquisition resulted in high-quality cardiac images which were free of major artifacts with spatial resolution of up to 0.21 × 0.21 × 1.00 mm(3) and a contrast-to-noise ratio (CNR) of 21 ± 3 between the myocardium and left ventricle. Self-gated golden ratio based radial acquisition successfully acquired cine images of the rat heart on a clinical MRI system without the need for dedicated animal ECG equipment.

In vivo micro-CT imaging of untreated and irradiated orthotopic glioblastoma xenografts in mice: capabilities, limitations and a comparison with bioluminescence imaging.

Small animal imaging is of increasing relevance in biomedical research. Studies systematically assessing the diagnostic accuracy of contrast-enhanced in vivo micro-CT of orthotopic glioma xenografts in mice do not exist. NOD/SCID/γc(-/-) mice (n = 27) underwent intracerebral implantation of 2.5 × 10(6) GFP-Luciferase-transduced U87MG cells. Mice underwent bioluminescence imaging (BLI) to detect tumor growth and afterwards repeated contrast-enhanced (300 µl Iomeprol i.v.) micro-CT imaging (80 kV, 75 µAs, 360° rotation, 1,000 projections, 33 s scan time, resolution 40 × 40 × 53 µm, 0.5 Gy/scan). Presence of tumors, tumor diameter and tumor volume in micro-CT were rated by two independent readers. Results were compared with histological analyses. Six mice with tumors confirmed by micro-CT received fractionated irradiation (3 × 5 Gy every other day) using the micro-CT (5 mm pencil beam geometry). Repeated micro-CT scans were tolerated well. Tumor engraftment rate was 74 % (n = 20). In micro-CT, mean tumor volume was 30 ± 33 mm(3), and the smallest detectable tumor measured 360 × 620 µm. The inter-rater agreement (n = 51 micro-CT scans) for the item tumor yes/no was excellent (Spearman-Rho = 0.862, p < 0.001). Sensitivity and specificity of micro-CT were 0.95 and 0.71, respectively (PPV = 0.91, NPV = 0.83). BLI on day 21 after tumor implantation had a sensitivity and specificity of 0.90 and 1.0, respectively (PPV = 1.0, NPV = 0.5). Maximum tumor diameter and volume in micro-CT and histology correlated excellently (tumor diameter: 0.929, p < 0.001; tumor volume: 0.969, p < 0.001, n = 17). Irradiated animals showed a large central tumor necrosis. Longitudinal contrast enhanced micro-CT imaging of brain tumor growth in live mice is feasible at high sensitivity levels and with excellent inter-rater agreement and allows visualization of radiation effects.

Magnetic resonance imaging signs of high intraventricular pressure--comparison of findings in dogs with clinically relevant internal hydrocephalus and asymptomatic dogs with ventriculomegaly.

BACKGROUND: Magnetic resonance imaging (MRI) findings of canine brains with enlarged ventricles in asymptomatic dogs were compared to those in dogs with clinically relevant internal hydrocephalus, in order to determine the imaging findings indicative of a relevant increase in intraventricular pressure. Discrimination between clinically relevant hydrocephalus and ventriculomegaly based on MRI findings has not been established yet and is anything but trivial because of the wide variation in ventricular size in different dog breeds and individuals. The MRI scans of the brains of 67 dogs of various breeds, skull conformation and weight were reviewed retrospectively. Based on clinical and imaging findings, the dogs were divided into three groups: a normal group (n = 20), a group with clinically silent ventriculomegaly (n = 25) and a group with severe clinically relevant internal hydrocephalus (n = 22). In addition to the ventricle/brain-index, a number of potential subjective signs of increased intraventricular pressure were recorded and compared between the groups. RESULTS: The ventricle/brain-index was significantly higher in dogs with relevant hydrocephalus (p < 0.001) and a threshold value of 0.6 was specified as a discriminator between internal hydrocephalus and ventriculomegaly. Other MR imaging findings associated with clinically relevant hydrocephalus were an elevation of the corpus callosum (p < 0.01), dorsoventral flattening of the interthalamic adhesion (p < 0.0001), periventricular edema (p < 0.0001), dilation of the olfactory recesses (p < 0.0001), thinning of the cortical sulci (p < 0.0001) and/or the subarachnoid space (p < 0.0027) and disruption of the internal capsule adjacent to the caudate nucleus (p < 0.0001). CONCLUSION: A combination of the abovementioned criteria may support a diagnosis of hydrocephalus that requires treatment.

Extrahepatic biliary atresia in a 4-week-old Pug.

OBJECTIVE: To report diagnosis and surgical therapy of extrahepatic biliary atresia in a dog. STUDY DESIGN: Clinical report. ANIMAL: Dog (n = 1). METHODS: A 4-week-old female intact Pug presented with acholic feces, lethargy, and poor weight gain. On the basis of the stool color and abdominal ultrasonography findings, extrahepatic biliary atresia was suspected. Intraoperatively, no major duodenal papilla was identified and cholecystoduodenostomy was performed. RESULTS: One day postoperatively the stool color was considered normal. On ultrasonographic examinations up to 6 months after surgery, the gall bladder remained small and the stoma was functional. Serum biochemical profile after 10 months had a mild increase in alanine-aminotransferase (149 U/L; reference interval [RI], 0-85 U/L) whereas alkaline phosphatase and glutamate dehydrogenase were within normal limits. At 15 months, clinical examination was normal and blood ammonia concentration was within normal limits (43 µmol/L; RI: <100 µmol/L) but bile acid concentrations were still markedly elevated (fasting, 95 µmol/L [RI: <20 µmol/L] to postprandial, 127 µmol/L [RI: <35 µmol/L]). CONCLUSIONS: Ultrasonography and stool color can be helpful noninvasive diagnostic evidence in extrahepatic biliary atresia. Surgical correction with cholecystoduodenostomy with a good clinical outcome is possible even in very young dogs. Long-term prognosis in dogs with extrahepatic biliary atresia is unknown.

Focal bacterial meningitis following ascending bite wound infection leading to paraparesis in a captive California sea lion (Zalophus californianus).

Magnetic resonance imaging was performed on a 15-yr-old captive female California sea lion (Zalophus californianus) with a 2-wk history of progressive paraparesis and a 9-mo history of exudative skin lesion on the left thoracic wall. Magnetic resonance images showed a well-defined muscle infiltrating lesion ventrolateral to the seventh cervical to the third thoracic vertebra on the left side, which extended through the left intervertebral foramina C7 to T3 into the vertebral canal, causing spinal cord compression and displacement as well as inflammation of the spinal cord and nerves. This lesion surprisingly caused no forelimb deficits. Differential diagnoses included abscess formation or neoplasia. Pathologic examination revealed chronic focal purulent meningitis associated with widespread paraspinal fistulous inflammation originating from a chronic dermal ulcer. Mainly Escherichia coli var. haemolytica and Clostridium perfringens were identified as the underlying agents.

Retrospective reconstruction of cardiac cine images from golden-ratio radial MRI using one-dimensional navigators.

PURPOSE: To demonstrate radial golden-ratio-based cardiac cine imaging by using interspersed one-dimensional (1D) navigators. MATERIALS AND METHODS: The 1D navigators were interspersed into the acquisition of radial spokes which were continuously rotated by an angle increment based on the golden-ratio. Performing correlation analysis between the 1D navigator projections, time points corresponding to the same cardiac motion phases were automatically identified and used to combine retrospectively golden-ratio rotated radial spokes from multiple data windows. Data windows were shifted consecutively for dynamic reconstruction of different cardiac motion frames. Experiments were performed during a single breathhold. By artificially reducing the amount of input data, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) as well as artifact level was evaluated for different breathhold durations. RESULTS: Analysis of the 1D navigator data provided a detailed correlation function revealing cardiac motion over time. Imaging results were comparable to images reconstructed based on a timely synchronized ECG. Cardiac cine images with a low artifact level and good image quality in terms of SNR and CNR were reconstructed from volunteer data achieving a CNR between the myocardium and the left ventricular cavity of 50 for the longest breathhold duration of 26 s. CNR maintained a value higher than 30 for acquisition times as low as 10 s. CONCLUSION: Combining radial golden-ratio-based imaging with an intrinsic navigator is a promising and robust method for performing high quality cardiac cine imaging.