Optimizing airway wall segmentation and quantification by reducing the influence of adjacent vessels and intravascular contrast material with a modified integral-based algorithm in quantitative computed tomography.

INTRODUCTION: Quantitative analysis of multi-detector computed tomography (MDCT) plays an increasingly important role in assessing airway disease. Depending on the algorithms used, airway dimensions may be over- or underestimated, primarily if contrast material was used. Therefore, we tested a modified integral-based method (IBM) to address this problem. METHODS: Temporally resolved cine-MDCT was performed in seven ventilated pigs in breath-hold during iodinated contrast material (CM) infusion over 60s. Identical slices in non-enhanced (NE), pulmonary-arterial (PA), systemic-arterial (SA), and venous phase (VE) were subjected to an in-house software using a standard and a modified IBM. Total diameter (TD), lumen area (LA), wall area (WA), and wall thickness (WT) were measured for ten extra- and six intrapulmonary airways. RESULTS: The modified IBM significantly reduced TD by 7.6%, LA by 12.7%, WA by 9.7%, and WT by 3.9% compared to standard IBM on non-enhanced CT (p<0.05). Using standard IBM, CM led to a decrease of all airway parameters compared to NE. For example, LA decreased from 80.85±49.26mm2 at NE, to 75.14±47.96mm2 (-7.1%) at PA (p<0.001), 74.96±48.55mm2 (-7.3%) at SA (p<0.001), and to 78.95±48.94mm2 (-2.4%) at VE (p = 0.200). Using modified IBM, the differences were reduced to -3.1% at PA, -2.9% at SA and -0.7% at VE (p<0.001; p<0.001; p = 1.000). CONCLUSIONS: The modified IBM can optimize airway wall segmentation and reduce the influence of CM on quantitative CT. This allows a more precise measurement as well as potentially the comparison of enhanced with non-enhanced scans in inflammatory airway disease.

Mind the gut: genomic insights to population divergence and gut microbial composition of two marine keystone species.

BACKGROUND: Deciphering the mechanisms governing population genetic divergence and local adaptation across heterogeneous environments is a central theme in marine ecology and conservation. While population divergence and ecological adaptive potential are classically viewed at the genetic level, it has recently been argued that their microbiomes may also contribute to population genetic divergence. We explored whether this might be plausible along the well-described environmental gradient of the Baltic Sea in two species of sand lance (Ammodytes tobianus and Hyperoplus lanceolatus). Specifically, we assessed both their population genetic and gut microbial composition variation and investigated not only which environmental parameters correlate with the observed variation, but whether host genome also correlates with microbiome variation. RESULTS: We found a clear genetic structure separating the high-salinity North Sea from the low-salinity Baltic Sea sand lances. The observed genetic divergence was not simply a function of isolation by distance, but correlated with environmental parameters, such as salinity, sea surface temperature, and, in the case of A. tobianus, possibly water microbiota. Furthermore, we detected two distinct genetic groups in Baltic A. tobianus that might represent sympatric spawning types. Investigation of possible drivers of gut microbiome composition variation revealed that host species identity was significantly correlated with the microbial community composition of the gut. A potential influence of host genetic factors on gut microbiome composition was further confirmed by the results of a constrained analysis of principal coordinates. The host genetic component was among the parameters that best explain observed variation in gut microbiome composition. CONCLUSIONS: Our findings have relevance for the population structure of two commercial species but also provide insights into potentially relevant genomic and microbial factors with regards to sand lance adaptation across the North Sea-Baltic Sea environmental gradient. Furthermore, our findings support the hypothesis that host genetics may play a role in regulating the gut microbiome at both the interspecific and intraspecific levels. As sequencing costs continue to drop, we anticipate that future studies that include full genome and microbiome sequencing will be able to explore the full relationship and its potential adaptive implications for these species.

Heavy-atom effect on optically excited triplet state kinetics.

In several fields of research, like e.g. photosensitization, photovoltaics, organic electroluminescent devices, dynamic nuclear polarization, or pulsed dipolar electron paramagnetic resonance spectroscopy, triplet state kinetics play an important role. It is therefore desirable to tailor the kinetics of photoexcited triplet states, e.g. by exploiting the intramolecular heavy-atom effect, and to determine the respective kinetic parameters. In this work, we set out to systematically investigate the photoexcited triplet state kinetics of a series of haloanthracenes by time-resolved electron paramagnetic resonance spectroscopy in combination with synchronized laser excitation. For this purpose, a procedure to simulate time traces by solving the differential equation system governing the triplet kinetics numerically is developed. This way, spin lattice relaxation rates and zero-field triplet life times are obtained concurrently by a global fit to experimental data measured at three different cryogenic temperatures.

Delocalization of Coherent Triplet Excitons in Linear Rigid Rod Conjugated Oligomers.

In this work, the triplet state delocalization in a series of monodisperse oligo(p-phenyleneethynylene)s (OPEs) is studied by pulsed electron paramagnetic resonance (EPR) and pulsed electron nuclear double resonance (ENDOR) determining zero-field splitting, optical spin polarization, and proton hyperfine couplings. Neither the zero-field splitting parameters nor the optical spin polarization change significantly with OPE chain length, in contrast to the hyperfine coupling constants, which showed a systematic decrease with chain length n according to a 2/(1 + n) decay law. The results provide striking evidence for the Frenkel-type nature of the triplet excitons exhibiting full coherent delocalization in the OPEs under investigation with up to five OPE repeat units and with a spin density distribution described by a nodeless particle in the box wave function. The same model is successfully applied to recently published data on π-conjugated porphyrin oligomers.

Photoexcited Triplet State Kinetics Studied by Electron Paramagnetic Resonance Spectroscopy.

Following the first evidence of the triplet character of the optically excited phosphorescent state of naphthalene by Hutchison and Mangum in 1958, electron paramagnetic resonance (EPR) spectroscopy has become widely used to study and understand the properties and kinetic characteristics of excited triplet states. This minireview gives an overview over EPR techniques based on continuous microwave methods using lock-in or direct detection as well as pulsed EPR methods with respect to their suitability for kinetic studies of excited triplet states. A short historical overview of the experimental and theoretical developments in this field of research as well as of the triplet systems studied, with a final focus on fullerenes, is given. This may help newcomers to the field as a guide to the relevant literature.

Laser-Induced Magnetic Dipole Spectroscopy.

Pulse electron paramagnetic resonance measurements of nanometer scale distance distributions have proven highly effective in structural studies. They exploit the magnetic dipole-dipole coupling between spin labels site-specifically attached to macromolecules. The most commonly applied technique is double electron-electron resonance (DEER, also called pulsed electron double resonance (PELDOR)). Here we present the new technique of laser-induced magnetic dipole (LaserIMD) spectroscopy based on optical switching of the dipole-dipole coupling. In a proof of concept experiment on a model peptide, we find, already at a low quantum yield of triplet excitation, the same sensitivity for measuring the distance between a porphyrin and a nitroxide label as in a DEER measurement between two nitroxide labels. On the heme protein cytochrome C, we demonstrate that LaserIMD allows for distance measurements between a heme prosthetic group and a nitroxide label, although the heme triplet state is not directly observable by an electron spin echo.

Pulmonary perfusion imaging: Qualitative comparison of TCIR MRI and SPECT/CT in porcine lung.

OBJECTIVES: To validate the anatomical accuracy, homogeneity and sensitivity of two-compartment modeled inversion recovery (TCIR) magnetic resonance imaging (MRI) in a multimodal animal experiment as a non-invasive alternative to standard functional imaging techniques. METHODS: Seven pigs were studied on a 1.5 T whole-body MR scanner and SPECT/CT. The specimens were intubated and maintained in general anesthesia throughout the experiment. TCIR maps of the fractional pulmonary blood volume were compared to dynamic contrast enhanced MRI and SPECT/CT via a region of interest (ROI) based reader study. A comprehensive statistical analysis was performed on the coefficient of variation to evaluate homogeneity properties. Sensitivity was assessed by detecting gravitation dependent perfusion variation and delineation of pathological areas. RESULTS: The fPBV-maps of all examined specimens indicate a superior homogeneity in the computed values (p<1.3×10(-4)). The sensitivity of the TCIR maps to a gravitation effect on the blood distribution was verified and a similar anteroposterior signal and count dependency was observed in DCE MRI and SPECT. Bland-Altman analysis showed no significant intra- or inter-observer difference within the ROI reader study (p>0.06). CONCLUSION: Superior information content, significantly higher homogeneity and similar sensitivity of TCIR when compared to DCE and SPECT/CT demonstrated the feasibility of TCIR MRI as an alternative contrast agent-free, non-invasive functional lung imaging approach.

Free breathing 1H MRI of the human lung with an improved radial turbo spin-echo.

OBJECTIVE: To optimize a radial turbo spin-echo sequence for motion-robust morphological lung magnetic resonance imaging (MRI) in free respiration. MATERIALS AND METHODS: A versatile multi-shot radial turbo spin-echo (rTSE) sequence is presented, using a modified golden ratio-based reordering designed to prevent coherent streaking due to data inconsistencies from physiological motion and the decaying signal. The point spread function for a moving object was simulated using a model for joint respiratory and cardiac motion with a concomitant T2 signal decay and with rTSE acquisition using four different reordering techniques. The reordering strategies were compared in vivo using healthy volunteers and the sequence was tested for feasibility in two patients with lung cancer and pneumonia. RESULTS: Simulations and in vivo measurements showed very weak artifacts, aside from motion blur, using the proposed reordering. Due to the opportunity for longer scan times in free respiration, a high signal-to-noise ratio (SNR) was achieved, facilitating identification of the disease as compared to standard half-Fourier-acquisition single-shot turbo spin-echo (HASTE) scans. Additionally, post-processing allowed modifying the T2 contrast retrospectively, further improving the diagnostic fidelity. CONCLUSION: The proposed radial TSE sequence allowed for high-resolution imaging with limited obscuring artifacts. The radial k-space traversal allowed for versatile post-processing that may help to improve the diagnosis of subtle diseases.

Monitoring enzymatic ATP hydrolysis by EPR spectroscopy.

An adenosine triphosphate (ATP) analogue modified with two nitroxide radicals is developed and employed to study its enzymatic hydrolysis by electron paramagnetic resonance spectroscopy. For this application, we demonstrate that EPR holds the potential to complement fluorogenic substrate analogues in monitoring enzymatic activity.

Clinical performance of the novel DiaSorin LIAISON(®) XL murex: HBsAg Quant, HCV-Ab, HIV-Ab/Ag assays.

BACKGROUND: The fully automated and closed LIAISON(®)XL platform was developed for reliable detection of infection markers like hepatitis B virus (HBV) surface antigen (HBsAg), hepatitis C virus (HCV) antibodies (Ab) or human immunodeficiency virus (HIV)-Ag/Ab. To date, less is known about the diagnostic performance of this system in direct comparison to the common Abbott ARCHITECT(®) platform. OBJECTIVES: We compared the diagnostic performance and usability of the DiaSorin LIAISON(®)XL with the commonly used Abbott ARCHITECT(®) system. STUDY DESIGN: The qualitative performance of the above mentioned assays was compared in about 500 sera. Quantitative tests were performed for HBsAg-positive samples from patients under therapy (n=289) and in vitro expressed mutants (n=37). For HCV-Ab, a total number of 155 selected samples from patients chronically infected with different HCV genotypes were tested. RESULTS: The concordance between both systems was 99.4% for HBsAg, 98.81% for HCV-Ab, and 99.6% for HIV-Ab/Ag. The quantitative LIAISON(®)XL murex HBsAg assay detected all mutants in comparable amounts to the HBsAg wild type and yielded highly reliable HBsAg kinetics in patients treated with antiviral drugs. Dilution experiments using the 2nd International Standard for HBsAg (WHO) showed a high accuracy of this test. HCV-Ab from patients infected with genotypes 1-3 were equally detected in both systems. Interestingly, S/CO levels of HCV-Ab from patients infected with genotype 3 seem to be relatively low using both systems. CONCLUSIONS: The LIAISON(®)XL platform proved to be an excellent system for diagnostics of HBV, HCV, and HIV with equal performance compared to the ARCHITECT(®) system.

DC-gated high resolution three-dimensional lung imaging during free-breathing.

PURPOSE: To use the acquisition of the k-space center signal (DC signal) implemented into a Cartesian three-dimensional (3D) FLASH sequence for retrospective respiratory self-gating and, thus, for the examination of the whole human lung in high spatial resolution during free breathing. MATERIALS AND METHODS: Volunteer as well as patient measurements were performed under free breathing conditions. The DC signal is acquired after the actual image data acquisition within each excitation of a 3D FLASH sequence. The DC signal is then used to track respiratory motion for retrospective respiratory gating. RESULTS: It is shown that the acquisition of the DC signal after the imaging module can be used in a 3D FLASH sequence to extract respiratory motion information for retrospective respiratory self-gating and allows for shorter echo times (TE) and therefore increased lung parenchyma SNR. CONCLUSION: The acquisition of the DC signal after image signal acquisition allows successful retrospective gating, enabling the reconstruction of high resolution images of the whole human lung under free breathing conditions.

Non-contrast-enhanced MRI of the pulmonary blood volume using two-compartment-modeled T1 -relaxation.

PURPOSE: To introduce a novel technique, based on a two-compartment model and nonselective inversion recovery (TCIR) for the non-contrast-enhanced evaluation of the fractional pulmonary blood volume (fPBV). MATERIALS AND METHODS: Ten healthy volunteers and one patient with focal lung destruction underwent examination with conventional magnetic resonance imaging (MRI) and TCIR. The reproducibility of TCIR was evaluated statistically, analyzing three consecutive measurement series. In order to evaluate the sensitivity of TCIR, the influence of gravitation on fPBV values along the anterior-posterior direction was assessed. Therefore, two transverse parameter maps of each volunteer in supine and prone position were acquired and analyzed. A comparison of patient images from TCIR-MRI, dynamic contrast-enhanced (DCE) MRI, and contrast-enhanced computed tomography (CT) was performed visually. RESULTS: The statistical evaluation showed significant similarity within the volunteer group, proving reproducibility. The detected slope of the fPBV values in anterior-posterior direction for both supine and prone position demonstrated the sensitivity of TCIR to the gravitational effect on the pulmonary blood distribution. The comparison between CT, DCE-, and TCIR-MRI patient datasets showed high similarity in dimension and location of the pathological part of the lung parenchyma. CONCLUSION: The introduced TCIR-technique is able to provide reproducible maps of the fPBV without the application of intravenous contrast media.

Locally resolved membrane binding affinity of the N-terminus of α-synuclein.

α-Synuclein is abundantly present in Lewy bodies, characteristic of Parkinson's disease. Its exact physiological role has yet to be determined, but mitochondrial membrane binding is suspected to be a key aspect of its function. Electron paramagnetic resonance spectroscopy in combination with site-directed spin labeling allowed for a locally resolved analysis of the protein-membrane binding affinity for artificial phospholipid membranes, supported by a study of binding to isolated mitochondria. The data reveal that the binding affinity of the N-terminus is nonuniform.

Morphologic and functional scoring of cystic fibrosis lung disease using MRI.

Magnetic resonance imaging (MRI) gains increasing importance in the assessment of cystic fibrosis (CF) lung disease. The aim of this study was to develop a morpho-functional MR-scoring-system and to evaluate its intra- and inter-observer reproducibility and clinical practicability to monitor CF lung disease over a broad severity range from infancy to adulthood. 35 CF patients with broad age range (mean 15.3 years; range 0.5-42) were examined by morphological and functional MRI. Lobe based analysis was performed for parameters bronchiectasis/bronchial-wall-thickening, mucus plugging, abscesses/sacculations, consolidations, special findings and perfusion defects. The maximum global score was 72. Two experienced radiologists scored the images at two time points (interval 10 weeks). Upper and lower limits of agreement, concordance correlation coefficients (CCC), total deviation index and coverage probability were calculated for global, morphology, function, component and lobar scores. Global scores ranged from 6 to 47. Intra- and inter-reader agreement for global scores were good (CCC: 0.98 (R1), 0.94 (R2), 0.97 (R1/R2)) and were comparable between high and low scores. Our results indicate that the proposed morpho-functional MR-scoring-system is reproducible and applicable for semi-quantitative evaluation of a large spectrum of CF lung disease severity. This scoring-system can be applied for the routine assessment of CF lung disease and maybe as endpoint for clinical trials.

The feasibility of low mechanical index contrast enhanced ultrasound (CEUS) in distinguishing malignant from benign thoracic lesions.

We proposed to assess the feasibility of low mechanical index (MI) contrast enhanced ultrasound (CEUS) in the characterisation of thoracic lesions. Fifty patients were prospectively examined by CEUS and images acquired on a low MI (0.17-0.24) setting following injection of SonoVue. From region-of-interest (ROI) generated signal intensity (SI) time curves, the maximum SI, bolus arrival time (BAT), time to peak intensity (TTP), wash-in slope and mean transit time (MTT) were calculated. Using the Wilcoxon rank test; parameters and threshold values for positive differentiation were determined. In addition, for the parameters that allowed positive differentiation between malignant and benign lesions receiver operator curves (ROC) were obtained. The wash-in slope, TTP and MTT (p = 0.0003, <0.0001, 0.02) allowed positive differentiation. The sensitivity and specificity was 93% and 78%, with 6.87 s(-1) threshold value for the wash-in slope, 78% and 89% with 11.84 s threshold for the TTP and 48% and 89% with 78.6 s threshold for the MTT. CEUS is a useful tool for differentiating malignant and benign thoracic lesions.

Isoelectric focusing pattern of plasminogen mutants of patients with hypoplasminogenemia: correlation of in-vitro data with computer-predicted isoelectric points (pI).

Plasminogen (plg), the circulating proenzyme of plasmin in blood, is a polymorphic protein and most of these natural variants have been identified using isoelectric focusing (IEF) gel electrophoresis. Here, we show that a rare plg gene polymorphism 504R/W is associated with IEF phenotype A3 on the protein level. One healthy individual with homozygous plg gene polymorphism 504W studied so far exhibited low normal plg antigen and slightly decreased plg activity, suggesting that this polymorphism is associated with (mild) hypoplasminogenemia. In addition, we present the findings of IEF phenotyping of plg mutants of 26 patients with severe hypoplasminogenemia, showing one of the following four IEF patterns: A3-like, A3A-like, B-like and AB-like. In the plasma of most compound heterozygous patients, only one of the two plg mutants was detectable by IEF electrophoresis, probably due to undetectable plasma concentration of the 2nd plg mutant. In almost all cases, pI of plg mutants and variants predicted by computer modeling were in good agreement with the observed IEF band pattern. plg phenotyping by IEF in combination with molecular genetic analysis of the plg gene is a useful approach to characterize plg mutants and variants further.

Pulmonary functional imaging: qualitative comparison of Fourier decomposition MR imaging with SPECT/CT in porcine lung.

PURPOSE: To compare unenhanced lung ventilation-weighted (VW) and perfusion-weighted (QW) imaging based on Fourier decomposition (FD) magnetic resonance (MR) imaging with the clinical reference standard single photon emission computed tomography (SPECT)/computed tomography (CT) in an animal experiment. MATERIALS AND METHODS: The study was approved by the local animal care committee. Lung ventilation and perfusion was assessed in seven anesthetized pigs by using a 1.5-T MR imager and SPECT/CT. For time-resolved FD MR imaging, sets of lung images were acquired by using an untriggered two-dimensional balanced steady-state free precession sequence (repetition time, 1.9 msec; echo time, 0.8 msec; acquisition time per image, 118 msec; acquisition rate, 3.33 images per second; flip angle, 75°; section thickness, 12 mm; matrix, 128 × 128). Breathing displacement was corrected with nonrigid image registration. Parenchymal signal intensity was analyzed pixelwise with FD to separate periodic changes of proton density induced by respiration and periodic changes of blood flow. Spectral lines representing respiratory and cardiac frequencies were integrated to calculate VW and QW images. Ventilation and perfusion SPECT was performed after inhalation of dispersed technetium 99m ((99m)Tc) and injection of (99m)Tc-labeled macroaggregated albumin. FD MR imaging and SPECT data were independently analyzed by two physicians in consensus. A regional statistical analysis of homogeneity and pathologic signal changes was performed. RESULTS: Images acquired in healthy animals by using FD MR imaging and SPECT showed a homogeneous distribution of VW and QW imaging and pulmonary ventilation and perfusion, respectively. The gravitation-dependent signal distribution of ventilation and perfusion in all animals was similarly observed at FD MR imaging and SPECT. Incidental ventilation and perfusion defects were identically visualized by using both modalities. CONCLUSION: This animal experiment demonstrated qualitative agreement in the assessment of regional lung ventilation and perfusion between contrast media-free and radiation-free FD MR imaging and conventional SPECT/CT.

Software-guided standardization of manual landmark data in medical images.

It is crucial to evaluate registration algorithms in order to make them available in clinical practice. Several evaluation strategies have been proposed in the past, and one approach is to evaluate these algorithms with intrinsic anatomical landmarks identified by a health professional. The acquisition and handling of large amounts of these landmark data is a time-consuming task for the health professional, and it is vulnerable to errors and inconsistencies. Additionally, limited access to appropriate tools makes dealing with landmark data considerably more difficult. We introduce a strategy for the acquisition of landmarks for the landmark-based evaluation of registration algorithms and we present an ontology-driven software tool that assists the different partners involved to act according to that strategy. This tool provides the user with intrinsic knowledge of the registration problems, the possibility to conveniently make the acquired data available to further processing, and an easy-to-use graphical interface.

Platelet Precursor Cells Can Be Generated from Cultured Human CD34+ Progenitor Cells But Display Recirculation into Hematopoietic Tissue upon Transfusion in Mice.

BACKGROUND: Whereas ex vivo expanded megakaryocytic progenitor cells have been investigated for their ability to support platelet regeneration, the question whether more mature platelet-like particles expanded from hematopoietic progenitor cells may be useful for transfusion purposes remains largely elusive. METHODS: Human peripheral blood progenitor cells (PBPCs) were enriched using surface expression of CD34 by immunoselection. CD34+ enriched PBPCs were expanded ex vivo in serum-free medium supplemented with cytokines. As a proof-of-principle, distribution of expanded CD61+ particles was analyzed after transfusion into Non-Obese Diabetic/ Severe Combined Immunodeficiency (NOD/SCID) mice. RESULTS: Highest ex vivo expansion for CD41+/CD61 + cells was achieved when medium was supplemented with SCF, TPO and IL-3. During expansion culture, CD34 marker expression decreased from 85 to 2-8%, while megakaryocytic cells appeared and CD41 and CD61 expression increased from 3 to about 30%. After transfusion of the expanded cells in NOD/SCID mice, CD61 + cells located mainly to bone marrow and to a lesser degree to spleen, but also circulated in blood. CONCLUSIONS: Platelet-like particles using cytokine-substituted serumfree medium can be generated efficiently from CD34+ expansion cultures, but mainly home to hematopoietic tissue.

Magnetic resonance imaging and computed tomography of respiratory mechanics.

Radiotherapy for organs with respiratory motion has motivated the development of dynamic volume lung imaging with computed tomography (4D-CT) or magnetic resonance imaging (4D-MRI). 4D-CT can be realized in helical (continuous couch translation during image acquisition) or cine mode (translation step-by-step), either acquired prospectively or reconstructed retrospectively with temporal resolutions of up to 250 msec. Long exposure times result in high radiation dose and restrict 4D-CT to specific indications (ie, radiotherapy planning). Dynamic MRI accelerated by parallel imaging and echo sharing reaches temporal resolutions of up to 10 images/sec (2D+t) or 1 volume/s (3D+t) that allow analyzing respiratory motion of the lung and its tumors. Near isotropic 4D-MRI can be used to assess tumor displacement, chest wall invasion, and segmental respiratory mechanics. Limited temporal resolution of dynamic volume acquisitions (in their current implementation) may lead to an overestimation of tumor size, as the mass is volume averaged into many voxels during motion. Nevertheless, 4D-MRI allows for repeated and prolonged measurements without radiation exposure and therefore appears to be appropriate for patient selection in motion-adapted radiotherapy as well as for a broad spectrum of scientific applications.