Dual-energy computed tomography quality control: Initial experiences with a semi-automatic analysis tool.

PURPOSE: A quality control (QC) system for dual-energy CT (DECT) was developed. The scope of the QC system was to monitor both the constancy of the CT images and the software used in calculating the DECT derived maps. Longitudinal analysis was based on a standard imaging protocol, a commercial multi-energy phantom, and a semi-automatic analysis tool. METHODS: The phantom consisted of an elliptical body section with round slots for interchangeable inserts. It was scanned with 90kVp/Sn150kVp, automatic tube current modulation, and 9.6 mGy CTDIvol. From the two conventional CT images, scanner manufacturer's software was used to provide virtual monoenergetic images at two different energies, effective atomic number (Zeff) maps, and iodine concentration maps. The images were analyzed using an open-source tool allowing user-selected statistics of interest. The means and standard deviations of the phantom background and the iodine, calcium, and water inserts were recorded. The QC tool is available at github.com/tomakela/dectqatool. RESULTS: The obtained results were generally highly consistent over time, except for the smaller diameter iodine inserts. A small change inZeff was observed after a DECT software update. The developed QC tool aided the analysis robustness: the segmentations were modifiable when needed, and small rotations or air bubbles in the water insert were easily corrected. CONCLUSION: The developed QC system provided easy-to-use workflow for constancy measurements. A small deviation due to change in the post-processing was detected. The proposed imaging protocol and analysis steps, and the reported measurement variations can aid in determining action levels for DECT QC.

Automatic detection of Crohn's disease using quantified motility in magnetic resonance enterography: initial experiences.

AIM: To report initial experiences of automatic detection of Crohn's disease (CD) using quantified motility in magnetic resonance enterography (MRE). MATERIALS AND METHODS: From 302 patients, three datasets with roughly equal proportions of CD and non-CD cases with various illnesses were drawn for testing and neural network training and validation. All datasets had unique MRE parameter configurations and were performed in free breathing. Nine neural networks were devised for automatic generation of three different regions of interests (ROI): small bowel, all bowel, and non-bowel. Additionally, a full-image ROI was tested. The motility in an MRE series was quantified via a registration procedure, which, accompanied with given ROIs, resulted in three motility indices (MI). A subset of the indices was used as an input for a binary logistic regression classifier, which predicted whether the MRE series represented CD. RESULTS: The highest mean area under the curve (AUC) score, 0.78, was reached using the full-image ROI and with the dataset with the highest cine series length. The best AUC scores for the other two datasets were only 0.54 and 0.49. CONCLUSION: The automatic system was able to detect CD in the group of MRE studies with lower temporal resolution and longer cine series showing potential in primary bowel disorder diagnostics. Larger ROI selections and utilising all available cine series for motility registration yielded slight performance improvements.

Fetal radiation dose in three common CT examinations during pregnancy - Monte Carlo study.

PURPOSE: To determine fetal doses in different stages of pregnancy in three common computed tomography (CT) examinations: pulmonary CT angiography, abdomino-pelvic and trauma scan with Monte Carlo (MC) simulations. METHODS: An adult female anthropomorphic phantom was scanned with a 64-slice CT using pulmonary angiography, abdomino-pelvic and trauma CT scan protocols. Three different sized gelatin boluses placed on the phantom's abdomen simulated different stages of pregnancy. Intrauterine dose was used as a surrogate to a dose absorbed to the fetus. MC simulations were performed to estimate uterine doses. The simulation dose levels were calibrated with volumetric CT dose index (CTDIvol) measurements and MC simulations in a cylindrical CTDI body phantom and compared with ten point doses measured with metal-oxide-semiconductor field-effect-transistor dosimeters. Intrauterine volumes and uterine walls were segmented and the respective dose volume histograms were calculated. RESULTS: The mean intrauterine doses in different stages of pregnancy varied from 0.04 to 1.04mGy, from 4.8 to 5.8mGy, and from 9.8 to 12.6mGy in the CT scans for pulmonary angiography, abdomino-pelvic and trauma CT scans, respectively. MC simulations showed good correlation with the MOSFET measurement at the measured locations. CONCLUSIONS: The three studied examinations provided highly varying fetal doses increasing from sub-mGy level in pulmonary CT angiography to notably higher levels in abdomino-pelvic and trauma scans where the fetus is in the primary exposure range. Volumetric dose distribution offered by MC simulations in an appropriate anthropomorphic phantom provides a comprehensive dose assessment when applied in adjunct to point-dose measurements.

Ion pair complexes and anion binding in the solution of a ditopic receptor.

The synthesis and crystal structures with alkali halides of a ditopic benzo-15-crown-5 bis-urea receptor have been presented. In addition, the anion binding properties of and its alkali metal complexes in solution are presented. A comprehensive single-crystal X-ray crystallographic study of , all together 13 crystal structures, including the ion pair complexes with NaCl, NaBr, NaI, KF, KCl, KBr, KI, RbF, RbCl, and RbI, give a detailed view of how behaves in the solid-state with different alkali halides depending on the size of the cation and anion. In the solid-state forms a 1 : 1 complex with a sodium cation and the anion is complexed as a contact (NaCl) or a separate ion pair (NaBr, NaI). With larger potassium and rubidium cations assembles into a 2 : 1 complex and forms a separated ion pair complex with the anion. Reflecting the crystal structures the forms a 1 : 1 complex with Na(+) in solution, and a 2 : 1 complex with K(+), which were verified by Job's plot analysis in 4 : 1 CDCl3/dimethyl sulfoxide. The binding strength of the monomeric [·Na](+) and the dimeric [2·K](+) toward chloride, bromide and iodide anions was studied by (1)H NMR titrations in 4 : 1 CDCl3/DMSO, and a clear turn-on effect of the cation complexation compared to the neutral receptor alone (Ka with for Cl(-), Br(-) and I(-) being 832, 174 and 32 M(-1), respectively) was observed. The monomeric [·Na](+) binds chloride 9, bromide 8, and iodide 12 times stronger than , while for the dimeric [2K](+) the corresponding increase in binding is 51 (Cl(-)), 84 (Br(-)), and 22 (I(-)) times with the same stoichiometric ratios as observed for the ion pair complexes in the solid-state.

Comparison of current density viability imaging at rest with FDG-PET in patients after myocardial infarction.

The assessment of myocardial viability is a major diagnostic challenge in patients with coronary artery disease (CAD) after myocardial infarction. Novel threedimensional current density (CD) imaging algorithms use high-resolution magnetic field mapping to determine the electrical activity of myocardial segments at rest. We, for the first time, compared CD activity obtained with several algorithms to 18-F-fluoro-deoxyglucose positron emission tomography (FDG-PET) in evaluation of myocardial viability. Magnetic field maps were obtained in nine adult patients (pt) with CAD and a history of infarction. The criterion for non-viable myocardium was an FDG-PET uptake with less than 45% of the maximum in the respective segments. CD imaging was applied to the left ventricle by using six different methods to solve the inverse problem. Mean CD activity was calculated for a close meshed grid of 90 locations of the left ventricle. A cardiologist compared bull's eye plots of CD and FDG-PET activity by eye. Spearman's correlation coefficients and specificity at a given level of sensitivity (70%) were calculated. Bull's eye plots revealed a significant correlation of CD/PET in 5 pt and no correlation in 3 pt. One pt had a negative correlation. The six different CD reconstruction methods performed similar. While CD reconstruction has the principal potential to image viable myocardium, we found that the reconstructed CD magnitude was low in scar segments but also reduced in some segments with preserved metabolic activity under resting conditions. New vector measurement techniques, the use of additional stress testing and advances in mathematical methodology are expected to improve CD imaging in future.

Mutation analysis of three genes encoding novel LKB1-interacting proteins, BRG1, STRADalpha, and MO25alpha, in Peutz-Jeghers syndrome.

Mutations in LKB1 lead to Peutz-Jeghers syndrome (PJS). However, only a subset of PJS patients harbours LKB1 mutations. We performed a mutation analysis of three genes encoding novel LKB1-interacting proteins, BRG1, STRADalpha, and MO25alpha, in 28 LKB1-negative PJS patients. No disease-causing mutations were detected in the studied genes in PJS patients from different European populations.

CAK-independent activation of CDK6 by a viral cyclin.

In normal cells, activation of cyclin-dependent kinases (cdks) requires binding to a cyclin and phosphorylation by the cdk-activating kinase (CAK). The Kaposi's sarcoma-associated herpesvirus encodes a protein with similarity to D-type cyclins. This KSHV-cyclin activates CDK6, alters its substrate specificity, and renders CDK6 insensitive to inhibition by the cdk inhibitor p16(INK4a). Here we investigate the regulation of the CDK6/KSHV-cyclin kinase with the use of purified proteins and a cell-based assay. We find that KSHV-cyclin can activate CDK6 independent of phosphorylation by CAK in vitro. In addition, CAK phosphorylation decreased the p16(INK4a) sensitivity of CDK6/KSHV-cyclin complexes. In cells, expression of CDK6 or to a lesser degree of a nonphosphorylatable CDK6(T177A) together with KSHV-cyclin induced apoptosis, indicating that CDK6 activation by KSHV-cyclin can proceed in the absence of phosphorylation by CAK in vivo. Coexpression of p16 partially protected cells from cell death. p16 and KSHV-cyclin can form a ternary complex with CDK6 that can be detected by binding assays as well as by conformational changes in CDK6. The Kaposi's sarcoma-associated herpesvirus has adopted a clever strategy to render cell cycle progression independent of mitogenic signals, cdk inhibition, or phosphorylation by CAK.

Vascular abnormalities and deregulation of VEGF in Lkb1-deficient mice.

The LKB1 tumor suppressor gene, mutated in Peutz-Jeghers syndrome, encodes a serine/threonine kinase of unknown function. Here we show that mice with a targeted disruption of Lkb1 die at midgestation, with the embryos showing neural tube defects, mesenchymal cell death, and vascular abnormalities. Extraembryonic development was also severely affected; the mutant placentas exhibited defective labyrinth layer development and the fetal vessels failed to invade the placenta. These phenotypes were associated with tissue-specific deregulation of vascular endothelial growth factor (VEGF) expression, including a marked increase in the amount of VEGF messenger RNA. Moreover, VEGF production in cultured Lkb1(-/-) fibroblasts was elevated in both normoxic and hypoxic conditions. These findings place Lkb1 in the VEGF signaling pathway and suggest that the vascular defects accompanying Lkb1 loss are mediated at least in part by VEGF.

alpha,beta-Dibenzyl-gamma-butyrolactone lignan alcohols: total synthesis of (+/-)-7'-hydroxyenterolactone, (+/-)-7'-hydroxymatairesinol and (+/-)-8-hydroxyenterolactone.

Two trans-alpha,beta-dibenzyl-gamma-butyrolactone lignans carrying a hydroxyl group at the beta-benzylic carbon atom and a alpha-hydroxy alpha,beta-dibenzyl-gamma-butyrolactone lignan were synthesized in racemic form using the tandem conjugate addition reaction to construct the basic lignan skeleton. Subsequent reaction steps involved either a catalytic reduction of the regenerated keto group to the alcohol, or a hydrogenolysis to benzylic methylene followed by lactone enolate formation and oxidation to give the alpha-hydroxybutyrolactones. These procedures were applied for the synthesis of 7'-hydroxyenterolactones and 7'-hydroxymatairesinols, and 8-hydroxyenterolactones, respectively. The diastereomeric mixtures of these compounds were separated either by HPLC techniques or column chromatography and the structures were elucidated using NMR spectroscopy.

Inability to enter S phase and defective RNA polymerase II CTD phosphorylation in mice lacking Mat1.

The trimeric Cdk7-cyclin H-Mat1 complex comprises the kinase subunit of basal transcription factor TFIIH and has been shown to function as a cyclin-dependent kinase (Cdk)-activating kinase. Herein we report that disruption of the murine Mat1 gene leads to peri-implantation lethality coincident with depletion of maternal Mat1 protein. In culture, Mat1(-/-) blastocysts gave rise to viable post-mitotic trophoblast giant cells while mitotic lineages failed to proliferate and survive. In contrast to wild-type trophoblast giant cells, Mat1(-/-) cells exhibited a rapid arrest in endoreduplication, which was characterized by an inability to enter S phase. Additionally, Mat1(-/-) cells exhibited defects in phosphorylation of the C-terminal domain (CTD) of RNA polymerase II on both Ser5 and Ser2 of the heptapeptide repeat. Despite this, Mat1(-/-) cells demonstrated apparent transcriptional and translational integrity. These data indicate an essential role for Mat1 in progression through the endocycle and suggest that while Mat1 modulates CTD phosphorylation, it does not appear to be essential for RNA polymerase II-mediated transcription.

BRCT domain-containing protein TopBP1 functions in DNA replication and damage response.

Topoisomerase IIbeta-binding protein (TopBP1), a human protein with eight BRCT domains, is similar to Saccharomyces cerevisiae Dpb11 and Schizosaccharomyces pombe Cut5 checkpoint proteins and closely related to Drosophila Mus101. We show that human TopBP1 is required for DNA replication and that it interacts with DNA polymerase epsilon. In S phase TopBP1 colocalizes with Brca1 to foci that do not represent sites of ongoing DNA replication. Inhibition of DNA synthesis leads to relocalization of TopBP1 together with Brca1 to replication forks, suggesting a role in rescue of stalled forks. DNA damage induces formation of distinct TopBP1 foci that colocalize with Brca1 in S phase, but not in G(1) phase. We also show that TopBP1 interacts with the checkpoint protein hRad9. Thus, these results implicate TopBP1 in replication and checkpoint functions.

Developmentally regulated expression of Smad3, Smad4, Smad6, and Smad7 involved in TGF-beta signaling.

Transforming growth factor-beta (TGF-beta) signaling is mediated from serine/threonine kinase receptors to transcriptional responses via Smad proteins. Here comparison of mRNA expression of Smad3-7 in mouse embryos (E9-E15) revealed developmentally regulated distinct expression patterns for Smad3, 4, 6, and 7. Smad3 was prominently expressed in the differentiating (from E10) central nervous system, but also in developing bones, branchial arches and epithelium of various tissues. Smad4 mostly showed ubiquitous expression, but in E15 embryos, a pronounced signal appeared in epithelial crypts of the gut. Inhibitory Smad6 and Smad7 were coexpressed at high levels in developing cardiovascular system from the earliest stages studied. In contrast, Smad6 was selectively expressed at high levels, e.g. in intramembranous bone whereas Smad7 was prominent in seminiferous tubules of the testis, demonstrating distinct expression of these genes in non-cardiovascular tissues.

Specificity of Cdk activation in vivo by the two Caks Mcs6 and Csk1 in fission yeast.

Activating phosphorylation of cyclin-dependent kinases (Cdks) is mediated by at least two structurally distinct types of Cdk-activating kinases (Caks): the trimeric Cdk7-cyclin H-Mat1 complex in metazoans and the single-subunit Cak1 in budding yeast. Fission yeast has both Cak types: Mcs6 is a Cdk7 ortholog and Csk1 a single-subunit kinase. Both phosphorylate Cdks in vitro and rescue a thermosensitive budding yeast CAK1 strain. However, this apparent redundancy is not observed in fission yeast in vivo. We have identified mutants that exhibit phenotypes attributable to defects in either Mcs6-activating phosphorylation or in Cdc2-activating phosphorylation. Mcs6, human Cdk7 and budding yeast Cak1 were all active as Caks for Cdc2 when expressed in fission yeast. Although Csk1 could activate Mcs6, it was unable to activate Cdc2. Biochemical experiments supported these genetic results: budding yeast Cak1 could bind and phosphorylate Cdc2 from fission yeast lysates, whereas fission yeast Csk1 could not. These results indicate that Mcs6 is the direct activator of Cdc2, and Csk1 only activates Mcs6. This demonstrates in vivo specificity in Cdk activation by Caks.

Current-density estimation of exercise-induced ischemia in patients with multivessel coronary artery disease.

Magnetocardiographic and body surface potential mapping data measured in 6 patients with multivessel coronary artery disease were used in equivalent current-density estimation (CDE). Patient-specific boundary-element torso models were acquired from magnetic resonance images. Positron emission tomography data registrated with anatomical magnetic resonance imaging data provided the gold standard. Discrete current-density estimation values were computed on the epicardial surface of the left ventricle from difference (stress-rest) ST-segment maps. The ill-posed inverse problem was regularized with 3 different methods (Tikhonov regularization with an identity or a surface Laplacian operator and a maximum a posteriori estimator). Comparisons with positron emission tomography studies showed that the maximum a posteriori estimator is superior to other regularizations, provided that a suitable a priori information is available. In general, good correspondence was found for segments of high and low amplitude in current-density estimations, and the viable and scar areas in positron emission tomography, respectively.

Intestinal metabolism of rye lignans in pigs.

To study the intestinal metabolism of lignans, the concentrations of plant and mammalian lignans in intestinal digesta sampled along the intestinal tract of pigs were determined by isotope dilution GC-MS. The pigs were fed rye-bread diets made from either whole rye-grains or rye-grain milling fractions enriched in pericarp-testa, aleurone or endosperm cells. The content and characteristics of dietary fibre varied between diets and had been shown to induce different colon fermentation patterns. As the metabolism of lignans depends on the action of the intestinal flora, we tested whether the rye-bread diets influence the metabolism of lignans. In the ileum, the lignans were mainly present as conjugated plant lignans, which were determined only when the analytical procedure included a hydrolysis step. High recovery of dietary lignans in the ileum may indicate that the lignans enter the enterohepatic circulation. In addition, two to three times the intake of lignans were recovered in the faeces when the diets had a high content of dietary fibre suggesting underestimation of plant lignans in the diet. Most of the plant lignans disappeared from the intestinal tract between the terminal ileum and the caecum. The intestinal concentrations and the disappearance of lignans correlated with the content of lignans in the diet, being highest on the pericarp-testa diet and lowest on the endosperm diet. No effect of fermentation pattern on the intestinal metabolism of lignans was observed. The lignans were liberated from the pericarp-testa diet although the plant cell walls remained largely undegraded.

Multimodality MR imaging assessment of myocardial viability: combination of first-pass and late contrast enhancement to wall motion dynamics and comparison with FDG PET-initial experience.

PURPOSE: To combine three magnetic resonance (MR) imaging modalities-dobutamine stress cine, first pass, and late contrast material-enhanced T1-weighted imaging-and to compare the results with 2-[fluorine 18]fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) in the assessment of unviable myocardium in coronary artery disease. MATERIALS AND METHODS: Ten patients with multivessel coronary artery disease underwent MR imaging before and 6 months after bypass surgery. Left ventricular cine MR imaging was performed at rest and during dobutamine infusion. Inversion-recovery gradient-echo images were obtained to study myocardial contrast enhancement at first pass and 5 minutes later. FDG PET was performed with orally administered acipimox before surgery. RESULTS: With dobutamine cine MR imaging, unviable myocardium was detected with a sensitivity of 79% and a specificity of 93%; postoperative wall thickening was the standard. First-pass analysis increased these values to 97% and 96%; analysis of late enhancement with T1-weighted imaging, to 62% and 98%. FDG PET had a sensitivity of 81% and a specificity of 86%. CONCLUSION: The combination of first-pass enhancement analysis and wall motion assessment with stress significantly increases the specificity of MR imaging in the detection of unviable sectors.

The apoptotic v-cyclin-CDK6 complex phosphorylates and inactivates Bcl-2.

v-cyclin encoded by Kaposi's sarcoma herpesvirus/human herpesvirus 8 (KSHV or HHV8) associates with cellular cyclin-dependent kinase 6 (CDK6) to form a kinase complex that promotes cell-cycle progression, but can also induce apoptosis in cells with high levels of CDK6. Here we show that whereas HHV8-encoded v-Bcl-2 protects against this apoptosis, cellular Bcl-2 has lost its anti-apoptotic potential as a result of an inactivating phosphorylation in its unstructured loop region. Moreover, we identify Bcl-2 as a new substrate for v-cyclin-CDK6 in vitro, and show that it is present in a complex with CDK6 in cell lysates. A Bcl-2 mutant with a S70A S87A double substitution in the loop region is not phosphorylated and provides resistance to apoptosis, indicating that inactivation of Bcl-2 by v-cyclin-CDK6 may be required for the observed apoptosis. Furthermore, the identification of phosphorylated Bcl-2 in HHV8-positive Kaposi's sarcoma indicates that HHV8-mediated interference with host apoptotic signalling pathways may encourage the development of Kaposi's sarcoma.

Interactions of Cdk7 and Kin28 with Hint/PKCI-1 and Hnt1 histidine triad proteins.

Cyclin-dependent kinase 7 (Cdk7) forms a trimeric complex with cyclin H and Mat1 to form the mammalian Cdk-activating kinase, CAK, as well as a part of the basal transcription factor TFIIH, where Cdk7 phosphorylates the C-terminal domain (CTD) of the large subunit of RNA polymerase II. Here, we report a novel interaction between Cdk7 and a histidine triad (HIT) family protein, Hint/PKCI-1. This interaction was initially observed in a yeast two-hybrid study and subsequently verified by co-immunoprecipitation and subcellular localization studies, where overexpression of Cdk7 leads to partial relocalization of Hint to the nucleus. The physical association is independent of cyclin H binding or Cdk7 kinase activity and is conserved between the related Sacharomyces cerevisiae CTD kinase Kin28 and the HIT protein Hnt1. Furthermore, combination of a disruption of HNT1 and a KIN28 temperature-sensitive allele in S. cerevisiae led to highly elongated cell morphology and reduced colony formation, indicating a genetic interaction between KIN28 and HNT1. The physical and genetic interactions of Hint and Hnt1 with Cdk7 and Kin28 suggest a role for this class of histidine triad proteins in the regulation of Cdk7 and Kin28 functions.

Synthesis of enterolactone and enterodiol precursors as potential inhibitors of human estrogen synthetase (aromatase).

A series of variably substituted derivatives of lignan lactones and diols were prepared using tandem conjugate addition reaction as a key step. These theoretical precursors of the mammalian lignans enterolactone 1 and enterodiol 3 are moderate or weak inhibitors of human aromatase activity.