Neutrophils seeking new neighbors: radiotherapy affects the cellular framework and the spatial organization in a murine breast cancer model.

Neutrophils are known to contribute in many aspects of tumor progression and metastasis. The presence of neutrophils or neutrophil-derived mediators in the tumor microenvironment has been associated with poor prognosis in several types of solid tumors. However, the effects of classical cancer treatments such as radiation therapy on neutrophils are poorly understood. Furthermore, the cellular composition and distribution of immune cells in the tumor is of increasing interest in cancer research and new imaging technologies allow to perform more complex spatial analyses within tumor tissues. Therefore, we aim to offer novel insight into intra-tumoral formation of cellular neighborhoods and communities in murine breast cancer. To address this question, we performed image mass cytometry on tumors of the TS/A breast cancer tumor model, performed spatial neighborhood analyses of the tumor microenvironment and quantified neutrophil-extracellular trap degradation products in serum of the mice. We show that irradiation with 2 × 8 Gy significantly alters the cellular composition and spatial organization in the tumor, especially regarding neutrophils and other cells of the myeloid lineage. Locally applied radiotherapy further affects neutrophils in a systemic manner by decreasing the serum neutrophil extracellular trap concentrations which correlates positively with survival. In addition, the intercellular cohesion is maintained due to radiotherapy as shown by E-Cadherin expression. Radiotherapy, therefore, might affect the epithelial-mesenchymal plasticity in tumors and thus prevent metastasis. Our findings underscore the growing importance of the spatial organization of the tumor microenvironment, particularly with respect to radiotherapy, and provide insight into potential mechanisms by which radiotherapy affects epithelial-mesenchymal plasticity and tumor metastasis.

iMPI: portable human-sized magnetic particle imaging scanner for real-time endovascular interventions.

Minimally invasive endovascular interventions have become an important tool for the treatment of cardiovascular diseases such as ischemic heart disease, peripheral artery disease, and stroke. X-ray fluoroscopy and digital subtraction angiography are used to precisely guide these procedures, but they are associated with radiation exposure for patients and clinical staff. Magnetic Particle Imaging (MPI) is an emerging imaging technology using time-varying magnetic fields combined with magnetic nanoparticle tracers for fast and highly sensitive imaging. In recent years, basic experiments have shown that MPI has great potential for cardiovascular applications. However, commercially available MPI scanners were too large and expensive and had a small field of view (FOV) designed for rodents, which limited further translational research. The first human-sized MPI scanner designed specifically for brain imaging showed promising results but had limitations in gradient strength, acquisition time and portability. Here, we present a portable interventional MPI (iMPI) system dedicated for real-time endovascular interventions free of ionizing radiation. It uses a novel field generator approach with a very large FOV and an application-oriented open design enabling hybrid approaches with conventional X-ray-based angiography. The feasibility of a real-time iMPI-guided percutaneous transluminal angioplasty (PTA) is shown in a realistic dynamic human-sized leg model.

Magnetic particle imaging for artifact-free imaging of intracranial flow diverter stents: A phantom study.

PURPOSE: Magnetic Particle Imaging (MPI) is a new, background- and radiation-free tomographic imaging method that enables near real-time imaging of superparamagnetic iron-oxide nanoparticles (SPIONs) with high temporal and spatial resolution. This phantom study aims to investigate the potential of MPI for visualization of the stent lumen in intracranial flow diverters (FD). METHODS: Nitinol FD of different dimensions (outer diameter: 3.5 mm, 4.0 mm, 5.5 mm; total length: 22-40 mm) were scanned in vascular phantoms in a custom-built MPI scanner (in-plane resolution: ~ 2 mm, field of view: 65 mm length, 29 mm diameter). Phantoms were filled with diluted (1:50) SPION tracer agent Ferucarbotran (10 µmol (Fe)/ml; NaCL). Each phantom was measured in 32 different projections (overall acquisition time per image: 3200 ms, 5averages). After image reconstruction from raw data, two radiologists assessed image quality using a 5-point Likert scale. The signal intensity profile was measured using a semi-automatic evaluation tool. RESULTS: MPI visualized the lumen of all FD without relevant differences between the stented vessel phantom and the reference phantom. At 3.5 mm image quality was slightly inferior to the larger diameters. The FD themselves neither generated an MPI signal nor did they lead to relevant imaging artifacts. Ratings of both radiologists showed no significant difference, interrater reliability was good (ICC 0.84). A quantitative evaluation of the signal intensity profile did not reveal any significant differences (p > 0.05) either. CONCLUSION: MPI visualizes the lumen of nitinol FD stents in vessel phantoms without relevant stent-induced artifacts.

Magnetic Particle Imaging for Quantification of Vascular Stenoses: A Phantom Study.

Magnetic particle imaging (MPI) is a promising new tomographic imaging method to detect the spatial distribution of superparamagnetic iron-oxide nanoparticles (SPIOs). The aim of this paper was to investigate the potential of MPI to quantify artificial stenoses in vessel phantoms. Custom-made stenosis phantoms (length 40 mm; inner diameter 8 mm) with different degrees of stenosis (0%, 25%, 50%, 75%, and 100%) were scanned in a custom-built MPI scanner (in-plane resolution: ~1-1.5 mm and field of view: 65 29 29 mm3). Phantoms were filled with diluted Feru-carbotran [SPIO agent, 5 mmol (Fe)/l]. Each measurement (overall acquisition time: 20 ms per image, 400 averages) was repeated ten times to assess reproducibility. The MPI signal was used for semi-automatic stenosis quantification. Two stenosis evaluation approaches were compared based on the signal intensity profile alongside the stenosis phantoms. Using a novel multi-step image evaluation approach, MPI allowed for accurate quantification of different stenosis grades. While low grade stenoses were slightly over-estimated, high grade stenoses were slightly underestimated. In particular, the 0%, 25%, and 50% stenosis phantoms revealed a 6.2% ± 0.8, 25.7% ± 1.0, and 48.0% ± 1.5 stenosis, respectively. The higher grade 75% stenosis phantom revealed a 73.3% ± 2.8 and the 100% stenosis phantom a 95.8%± 1.9 stenosis. MPI accurately visualized and quantified different stenosis grades in vessel phantoms with high reproducibility demonstrating its great potential for fast and radiation-free preclinical cardiovascular imaging.

First in vivo traveling wave magnetic particle imaging of a beating mouse heart.

Magnetic particle imaging (MPI) is a non-invasive imaging modality for direct detection of superparamagnetic iron-oxide nanoparticles based on the nonlinear magnetization response of magnetic materials to alternating magnetic fields. This highly sensitive and rapid method allows both a quantitative and a qualitative analysis of the measured signal. Since the first publication of MPI in 2005 several different scanner concepts have been presented and in 2009 the first in vivo imaging results of a beating mouse heart were shown. However, since the field of view (FOV) of the first MPI-scanner only covers a small region several approaches and hardware enhancements were presented to overcome this issue and could increase the FOV on cost of acquisition speed. In 2014 an alternative scanner concept, the traveling wave MPI (TWMPI), was presented, which allows scanning an entire mouse-sized volume at once. In this paper the first in vivo imaging results using the TWMPI system are presented. By optimizing the trajectory the temporal resolution is sufficiently high to resolve the dynamic of a beating mouse heart.

Ultrasonographic fetal head position to predict mode of delivery: a systematic review and bivariate meta-analysis.

OBJECTIVE: We performed a systematic review to determine whether sonographic assessment of occipital position of the fetal head can contribute to the prediction of the mode of delivery. METHODS: We performed a systematic literature search of electronic databases from inception to May 2011. Two reviewers independently extracted data from the included studies. We used a bivariate model to estimate point estimates for sensitivity and specificity curves for the outcome Cesarean delivery. Eligible studies were cohort studies or cross-sectional studies that reported on both the position of the fetal head, as assessed by ultrasound, before or at the beginning of active labor as well as the outcome of labor in women at term. RESULTS: We included 11 primary articles reporting on 5053 women, of whom 898 had a Cesarean section. All studies indicated disappointing values for sensitivity and specificity in the prediction of Cesarean section. Summary point estimates of sensitivity and specificity were 0.39 (95% CI, 0.32-0.48) and 0.71 (95% CI, 0.67-0.74), respectively. CONCLUSION: Sonographic assessment of occipital position of the fetal head before delivery should not be used in the prediction of mode of delivery.

Optimized method for the determination of 25 polychlorinated biphenyls in water samples using stir bar sorptive extraction followed by thermodesorption-gas chromatography/mass spectrometry.

An optimized method using stir bar sorptive extraction (SBSE) for the determination of 25 polychlorinated biphenyls (PCBs) from water samples among them three of the most toxic coplanar PCBs (PCB 77, PCB 126 and PCB 169) was developed. Since the investigated PCBs comprise all steps of chlorination (from PCB 1 as monochlorobiphenyl to PCB 209 as decachlorobiphenyl) the results should be representative for the total class of the 209 PCB congeners. For 8 ml spiked water samples with 2 ml methanol addition and 2 h exposure time of stir bars recoveries between 28% (PCB 209) and 93% (PCB 1, PCB 52, PCB 77) were found. Detection limits between 0.05 ng/l and 0.15 ng/l were calculated for the combination of SBSE and thermodesorption-GC/MS. The procedure was applied to the investigation of groundwater and river water samples from the industrial region of Bitterfeld northern Leipzig, Germany.

NMR experiments for the sign determination of homonuclear scalar and residual dipolar couplings.

A modified version of the JHH-TOCSY experiment, 'signed COSY', is presented that allows the determination of the sign of residual dipolar 1H-1H coupling constants with respect to the sign of one-bond 1H-X coupling constants in linear three-spin systems X-1H-1H, where X = 13C or 15N. In contrast to the original JHH-TOCSY experiments, the signs of J(HH) couplings may be determined for CH2-CH2 moieties and for uniformly 13C/15N-labelled samples. In addition, sensitivity is enhanced, diagonal peaks are suppressed and cross peaks are observed only between directly coupled protons, as in a COSY spectrum.

Acetogenic isoquinoline alkaloids. CXII. Separation and identification of dimeric naphthylisoquinoline alkaloids by liquid chromatography coupled to electrospray ionization mass spectrometry.

The atropodiastereomeric dimeric naphthylisoquinoline alkaloids, michellamines A (1a), B (1b) and C (1c), together with their monomers, korupensamines A (2a) and B (2b), were investigated using electrospray ionization tandem mass spectrometry coupled to liquid chromatography (LC-ESI-MS-MS). From the spectra obtained, characteristic product ions were chosen to monitor the chromatographic separation achieved on an RP-18 column. Under acidic conditions required for chromatographic analysis, the monomeric alkaloids 2a and 2b yielded protonated molecules [M + H]+, while the dimers, the michellamines, exhibited doubly protonated [M + 2H]2+ molecules. In addition, the coeluting alkaloids 1b and 2b were identified unambiguously be means of tandem mass spectrometry. Thus, together with the retention times of the alkaloids, the product ion spectra allowed us the identification of michellamines in the presence of their presumed biogenetic monomeric precursors. Application of the HPLC-MS-MS method successfully proved the enzymatic formation of michellamine C (1c) by in vitro dimerization of korupensamine B (2b).

Characterization of enzymes from Ancistrocladus (Ancistrocladaceae) and Triphyophyllum (Dioncophyllaceae) catalyzing oxidative coupling of naphthylisoquinoline alkaloids to michellamines.

Peroxidase active preparations from three Ancistrocladus species and from Triphyophyllum peltatum have been partially purified. They catalyze the oxidative dimerization of korupensamines A and B to michellamines A and C, respectively, as well as the mixed coupling to michellamines A, B, and C. All of these enzymes consist of single polypeptides of approximately 65 kDa with peroxidase activity as monomers. They were characterized as soluble proteins predominantly localized in the leaf phloem of all species examined. Comparative studies with various naphthol precursors revealed an unexpected substrate specificity. Only korupensamines were dimerized by the enzymes, while other monomers, even if closely related, were not accepted as substrates. The coupling reactions described here represent the first direct synthesis of michellamines from korupensamines without previous protection of these precursors.