Acid-Sphingomyelinase Triggered Fluorescently Labeled Sphingomyelin Containing Liposomes in Tumor Diagnosis after Radiation-Induced Stress

In liposomal delivery, a big question is how to release the loaded material into the correct place. Here, we will test the targeting and release abilities of our sphingomyelin-consisting liposome. A change in release parameters can be observed when sphingomyelin-containing liposome is treated with sphingomyelinase enzyme. Sphingomyelinase is known to be endogenously released from the different cells in stress situations. We assume the effective enzyme treatment will weaken the liposome making it also leakier. To test the release abilities of the SM-liposome, we developed several fluorescence-based experiments. In in vitro studies, we used molecular quenching to study the sphingomyelinase enzyme-based release from the liposomes. We could show that the enzyme treatment releases loaded fluorescent markers from sphingomyelin-containing liposomes. Moreover, the release correlated with used enzymatic activities. We studied whether the stress-related enzyme expression is increased if the cells are treated with radiation as a stress inducer. It appeared that the radiation caused increased enzymatic activity. We studied our liposomes' biodistribution in the animal tumor model when the tumor was under radiation stress. Increased targeting of the fluorescent marker loaded to our liposomes could be found on the site of cancer. The liposomal targeting in vivo could be improved by radiation. Based on our studies, we propose sphingomyelin-containing liposomes can be used as a controlled release system sensitive to cell stress

Molecular mechanisms in uterine epithelium during trophoblast binding: the role of small GTPase RhoA in human uterine Ishikawa cells

BACKGROUND: Embryo implantation requires that uterine epithelium develops competence to bind trophoblast to its apical (free) poles. This essential element of uterine receptivity seems to depend on a destabilisation of the apico-basal polarity of endometrial epithelium. Accordingly, a reorganisation of the actin cytoskeleton regulated by the small GTPase RhoA plays an important role in human uterine epithelial RL95-2 cells for binding of human trophoblastoid JAR cells. We now obtained new insight into trophoblast binding using human uterine epithelial Ishikawa cells. METHODS: Polarity of Ishikawa cells was investigated by electron microscopy, apical adhesiveness was tested by adhesion assay. Analyses of subcellular distribution of filamentous actin (F-actin) and RhoA in apical and basal cell poles were performed by confocal laser scanning microscopy (CLSM) with and without binding of JAR spheroids as well as with and without inhibition of small Rho GTPases by Clostridium difficile toxin A (toxin A). In the latter case, subcellular distribution of RhoA was additionally investigated by Western blotting. RESULTS: Ishikawa cells express apical adhesiveness for JAR spheroids and moderate apico-basal polarity. Without contact to JAR spheroids, significantly higher signalling intensities of F-actin and RhoA were found at the basal as compared to the apical poles in Ishikawa cells. RhoA was equally distributed between the membrane fraction and the cytosol fraction. Levels of F-actin and RhoA signals became equalised in the apical and basal regions upon contact to JAR spheroids. After inhibition of Rho GTPases, Ishikawa cells remained adhesive for JAR spheroids, the gradient of fluorescence signals of F-actin and RhoA was maintained while the amount of RhoA was reduced in the cytosolic fraction with a comparable increase in the membrane fraction. CONCLUSION: Ishikawa cells respond to JAR contact as well as to treatment with toxin A with rearrangement of F-actin and small GTPase RhoA but seem to be able to modify signalling pathways in a way not elucidated so far in endometrial cells. This ability may be linked to the degree of polar organisation observed in Ishikawa cells indicating an essential role of cell phenotype modification in apical adhesiveness of uterine epithelium for trophoblast in vivo

Imaging in the acute abdomen - part 1 Case examples of frequent organ-specific causes: liver, gallbladder, pancreas, spleen and vessels

The acute abdomen is characterized by acute abdominal pain with defensive muscular tension, can be triggered by a variety of diseases and sometimes represents a life-threatening condition. After clinical inspection, in most cases dedicated imaging should be performed immediately. The frequently causal appendicitis and cholecystitis can mostly be diagnosed with ultrasound. In other cases with unclear ultrasound findings or unclear severe symptoms, computer tomography (CT) is usually necessary without delay. In contrast, magnetic resonance imaging (MRI) is predominantly indicated in pregnant women and children with unclear ultrasound findings. Thus, the radiologist is an important gatekeeper in the diagnostics of acute abdomen. The radiologist should therefore be familiar with the correct imaging indications, the frequent and rare causes as well as the corresponding morphological imaging characteristics

Imaging in the acute abdomen-part 2 Case examples of frequent organ-specific causes: gastrointestinal tract and urogenital system

The acute abdomen is a potentially life-threatening condition and requires a rapid diagnosis. After clinical inspection and in cases with unclear ultrasound findings or unclear serious symptoms computed tomography (CT) and in pregnant women and children magnetic resonance imaging (MRI) is usually necessary. This second part of Imaging in the acute abdomen focuses on frequent organ specific causes of the gastrointestinal tract and the urogenital system

Fractal dimension of tumor microvasculature by dynamic contrast-enhanced ultrasound

Angiogenesis plays a fundamental role in the growth of several types of cancer. Characterization of the microvascular architecture has therefore gained attention as a possible tool for cancer localization. However, the available options to assess the main features characterizing the microvascular architecture require immunohistological analysis of the resected tumors. Contrast-enhanced ultrasound (CEUS) provides new options for minimally-invasive investigation of the microvasculature by spatiotemporal analysis of the ultrasound-contrast-agent (UCA) transport kinetics. In this work, we propose a novel method to analyze CEUS data. The fractal dimension (FD) of the flow distribution is employed as a geometrical feature to characterize the microvascular architecture. To this end, the fractal dimension of parametric maps reflecting blood flow, such as UCA wash-in rate and peak enhancement, is derived for areas representing different microvascular architectures. Subcutaneous xenograft models of DU-145 and PC-3 prostate-cancer lines in mice were employed as they show marked differences in the spatial distribution of the microvascular density (MVD) inside the tumor. The ability of the method to differentiate between these models was therefore evaluated. For validation purposes, comparison with immunohistological MVD and FD assessment, as well as with UCA dispersion maps, was also performed. The results show good agreement between FD and MVD assessment, with the proposed method able to differentiate between the two cancer lines

Quantitative accuracy of virtual non-contrast images derived from spectral detector computed tomography: an abdominal phantom study

Dual-energy CT allows for the reconstruction of virtual non-contrast (VNC) images. VNC images have the potential to replace true non-contrast scans in various clinical applications. This study investigated the quantitative accuracy of VNC attenuation images considering different parameters for acquisition and reconstruction. An abdomen phantom with 7 different tissue types (different combinations of 3 base materials and 5 iodine concentrations) was scanned using a spectral detector CT (SDCT). Different phantom sizes (S, M, L), volume computed tomography dose indices (CTDIvol 10, 15, 20 mGy), kernel settings (soft, standard, sharp), and denoising levels (low, medium, high) were tested. Conventional and VNC images were reconstructed and analyzed based on regions of interest (ROI). Mean and standard deviation were recorded and differences in attenuation between corresponding base materials and VNC was calculated (VNCerror). Statistic analysis included ANOVA, Wilcoxon test and multivariate regression analysis. Overall, the VNCerror was - 1.4 ± 6.1 HU. While radiation dose, kernel setting, and denoising level did not influence VNCerror significantly, phantom size, iodine content and base material had a significant effect (e.g. S vs. M: - 1.2 ± 4.9 HU vs. - 2.1 ± 6.0 HU; 0.0 mg/ml vs. 5.0 mg/ml: - 4.0 ± 3.5 HU vs. 5.1 ± 5.0 HU and 35-HU-base vs. 54-HU-base: - 3.5 ± 4.4 HU vs. 0.7 ± 6.5; all p ≤ 0.05). The overall accuracy of VNC images from SDCT is high and independent from dose, kernel, and denoising settings; however, shows a dependency on patient size, base material, and iodine content; particularly the latter results in small, yet, noticeable differences in VNC attenuation

Volumetric analysis of MRONJ lesions by semiautomatic segmentation of CBCT images

Purpose The purpose of this study was to evaluate potential differences in volumes of areas of osteolysis caused by medication-related osteonecrosis of the jaw (MRONJ) between the upper and lower jaw. We aim to analyze the clinical relevance of volumetric measurement of osteolytic lesions for surgical planning of MRONJ patients. Methods Sixty-seven patients who were clinically and histopathologically diagnosed with MRONJ were retrospectively included in this study. Cone beam computed tomography (CBCT) images were evaluated according to localization, affected anatomical structures, and volumetric measurement of osteolytic lesions caused by MRONJ in appliance of CBCT datasets by using ITK-SNAP. Results The most frequently affected localization is the mandible, whereas female patients show significantly more often lesions of the maxilla. The cortical bone was predominantly affected. Furthermore, the affection of teeth, sinus floor, inferior alveolar nerve canal, or even a pathological fracture of the mandible are infrequently existing. The volumetric measurements revealed a statistically significant greater absolute osteolysis volume in males. Conclusions Image analysis and volumetric measurements of osteolytic lesions of MRONJ patients is a helpful tool to further understand the clinical appearance and identify compromised anatomic landmarks. Volumetric analysis aids in pre-surgical planning and visualizes the individual extent of the disease for each patient

Fractal dimension of tumor microvasculature by dynamic contrast-enhanced ultrasound

Angiogenesis plays a fundamental role in the growth of several types of cancer. Characterization of the microvascular architecture has therefore gained attention as a possible tool for cancer localization. However, the available options to assess the main features characterizing the microvascular architecture require immunohistological analysis of the resected tumors. Contrast-enhanced ultrasound (CEUS) provides new options for minimally-invasive investigation of the microvasculature by spatiotemporal analysis of the ultrasound-contrast-agent (UCA) transport kinetics. In this work, we propose a novel method to analyze CEUS data. The fractal dimension (FD) of the flow distribution is employed as a geometrical feature to characterize the microvascular architecture. To this end, the fractal dimension of parametric maps reflecting blood flow, such as UCA wash-in rate and peak enhancement, is derived for areas representing different microvascular architectures. Subcutaneous xenograft models of DU-145 and PC-3 prostate-cancer lines in mice were employed as they show marked differences in the spatial distribution of the microvascular density (MVD) inside the tumor. The ability of the method to differentiate between these models was therefore evaluated. For validation purposes, comparison with immunohistological MVD and FD assessment, as well as with UCA dispersion maps, was also performed. The results show good agreement between FD and MVD assessment, with the proposed method able to differentiate between the two cancer lines