Short-Term Treatment with Bisphenol-A Leads to Metabolic Abnormalities in Adult Male Mice

Bisphenol-A (BPA) is one of the most widespread endocrine disrupting chemicals (EDC) used as the base compound in the manufacture of polycarbonate plastics. Although evidence points to consider exposure to BPA as a risk factor for insulin resistance, its actions on whole body metabolism and on insulin-sensitive tissues are still unclear. The aim of the present work was to study the effects of low doses of BPA in insulin-sensitive peripheral tissues and whole body metabolism in adult mice. Adult mice were treated with subcutaneous injection of 100 µg/kg BPA or vehicle for 8 days. Whole body energy homeostasis was assessed with in vivo indirect calorimetry. Insulin signaling assays were conducted by western blot analysis. Mice treated with BPA were insulin resistant and had increased glucose-stimulated insulin release. BPA-treated mice had decreased food intake, lower body temperature and locomotor activity compared to control. In skeletal muscle, insulin-stimulated tyrosine phosphorylation of the insulin receptor β subunit was impaired in BPA-treated mice. This impairment was associated with a reduced insulin-stimulated Akt phosphorylation in the Thr308 residue. Both skeletal muscle and liver displayed an upregulation of IRS-1 protein by BPA. The mitogen-activated protein kinase (MAPK) signaling pathway was also impaired in the skeletal muscle from BPA-treated mice. In the liver, BPA effects were of lesser intensity with decreased insulin-stimulated tyrosine phosphorylation of the insulin receptor β subunit

Amide proton transfer imaging of high intensity focused ultrasound-treated tumor tissue

Purpose: In this study, the suitability of amide proton transfer (APT) imaging as a biomarker for the characterization of high intensity focused ultrasound (HIFU)-treated tumor tissue was assessed. Methods: APT imaging was performed on tumor-bearing mice before (n=15), directly after (n=15) and at 3 days (n=8) after HIFU treatment. A control group (n=7) of nontreated animals was scanned at the same time points. Histogram analysis of the tumor APT-weighted signal distributions was performed to assess HIFU-induced changes in the tumor APT contrast. Results: Distinct regions of decreased APT-weighted signal were observed at both time points after HIFU treatment. Analysis of the tumor APT-weighted signal distribution showed a pronounced shift toward lower APT-weighted signal values after HIFU treatment. A significantly increased fraction of pixels with an APT-weighted signal value between-10 and-2% was observed both directly (0.37±0.16) and at 3 days (0.49±0.16) after HIFU treatment as compared to baseline (0.2260.16). Conclusion: The presented results show that APT imaging is sensitive to HIFU-induced changes in tumor tissue and may thus serve as a new biomarker for monitoring the response of tumor tissue to HIFU treatment

Automatic segmentation of subcutaneous mouse tumors by multiparametric MR analysis based on endogenous contrast

Object: Contrast-enhanced T1-weighted imaging is usually included in MRI procedures for automatic tumor segmentation. Use of an MR contrast agent may not be appropriate for some applications, however. We assessed the feasability of automatic tumor segmentation by multiparametric cluster analysis that uses intrinsic MRI contrast only. Materials and methods: Multiparametric MRI consisting of quantitative T1, T2, and apparent diffusion coefficient (ADC) mapping was performed in mice bearing subcutaneous tumors (n = 21). k-means and fuzzy c-means clustering with all possible combinations of MRI parameters, i.e. feature vectors, and 2–7 clusters were performed on the multiparametric data. Clusters associated with tumor tissue were selected on the basis of the relative signal intensity of tumor tissue in T2-weighted images. The optimum segmentation method was determined by quantitative comparison of automatic segmentation with manual segmentation performed by three observers. In addition, the automatically segmented tumor volumes from seven separate tumor data sets were quantitatively compared with histology-derived tumor volumes. Results: The highest similarity index between manual and automatic segmentation (SImanual,automatic = 0.82 ± 0.06) was observed for k-means clustering with feature vector {T2, ADC} and four clusters. A strong linear correlation between automatically and manually segmented tumor volumes (R2 = 0.99) was observed for this segmentation method. Automatically segmented tumor volumes also correlated strongly with histology-derived tumor volumes (R2 = 0.96). Conclusion: Automatic segmentation of mouse subcutaneous tumors can be achieved on the basis of endogenous MR contrast only

MRI methods for the evaluation of high intensity focused ultrasound tumor treatment: current status and future needs

Thermal ablation with high intensity focused ultrasound (HIFU) is an emerging noninvasive technique for the treatment of solid tumors. HIFU treatment of malignant tumors requires accurate treatment planning, monitoring and evaluation, which can be facilitated by performing the procedure in an MR-guided HIFU system. The MR-based evaluation of HIFU treatment is most often restricted to contrast-enhanced T1-weighted imaging, while it has been shown that the non-perfused volume may not reflect the extent of nonviable tumor tissue after HIFU treatment. There are multiple studies in which more advanced MRI methods were assessed for their suitability for the evaluation of HIFU treatment. While several of these methods seem promising regarding their sensitivity to HIFU-induced tissue changes, there is still ample room for improvement of MRI protocols for HIFU treatment evaluation. In this review article, we describe the major acute and delayed effects of HIFU treatment. For each effect, the MRI methods that have been - or could be - used to detect the associated tissue changes are described. In addition, the potential value of multiparametric MRI for the evaluation of HIFU treatment is discussed. The review ends with a discussion on future directions for the MRI-based evaluation of HIFU treatment

Cluster analysis of DCE-MRI data identifies regional tracer-kinetic changes after tumor treatment with high intensity focused ultrasound

Evaluation of high intensity focused ultrasound (HIFU) treatment with MRI is generally based on assessment of the non-perfused volume from contrast-enhanced T1-weighted images. However, the vascular status of tissue surrounding the non-perfused volume has not been extensively investigated with MRI. In this study, cluster analysis of the transfer constant K(trans) and extravascular extracellular volume fraction ve , derived from dynamic contrast-enhanced MRI (DCE-MRI) data, was performed in tumor tissue surrounding the non-perfused volume to identify tumor subregions with distinct contrast agent uptake kinetics. DCE-MRI was performed in CT26.WT colon carcinoma-bearing BALB/c mice before (n = 12), directly after (n = 12) and 3 days after (n = 6) partial tumor treatment with HIFU. In addition, a non-treated control group (n = 6) was included. The non-perfused volume was identified based on the level of contrast enhancement. Quantitative comparison between non-perfused tumor fractions and non-viable tumor fractions derived from NADH-diaphorase histology showed a stronger agreement between these fractions 3 days after treatment (R(2) to line of identity = 0.91) compared with directly after treatment (R(2) = 0.74). Next, k-means clustering with four clusters was applied to K(trans) and ve parameter values of all significantly enhanced pixels. The fraction of pixels within two clusters, characterized by a low K(trans) and either a low or high ve , significantly increased after HIFU. Changes in composition of these clusters were considered to be HIFU induced. Qualitative H&E histology showed that HIFU-induced alterations in these clusters may be associated with hemorrhage and structural tissue disruption. Combined microvasculature and hypoxia staining suggested that these tissue changes may affect blood vessel functionality and thereby tumor oxygenation. In conclusion, it was demonstrated that, in addition to assessment of the non-perfused tumor volume, the presented methodology gives further insight into HIFU-induced effects on tumor vascular status. This method may aid in assessment of the consequences of vascular alterations for the fate of the tissu

Automatic segmentation of subcutaneous mouse tumors by multiparametric MR analysis based on endogenous contrast

\u3cp\u3eObject: Contrast-enhanced T\u3csub\u3e1\u3c/sub\u3e-weighted imaging is usually included in MRI procedures for automatic tumor segmentation. Use of an MR contrast agent may not be appropriate for some applications, however. We assessed the feasability of automatic tumor segmentation by multiparametric cluster analysis that uses intrinsic MRI contrast only. Materials and methods: Multiparametric MRI consisting of quantitative T\u3csub\u3e1\u3c/sub\u3e, T\u3csub\u3e2\u3c/sub\u3e, and apparent diffusion coefficient (ADC) mapping was performed in mice bearing subcutaneous tumors (n = 21). k-means and fuzzy c-means clustering with all possible combinations of MRI parameters, i.e. feature vectors, and 2–7 clusters were performed on the multiparametric data. Clusters associated with tumor tissue were selected on the basis of the relative signal intensity of tumor tissue in T\u3csub\u3e2\u3c/sub\u3e-weighted images. The optimum segmentation method was determined by quantitative comparison of automatic segmentation with manual segmentation performed by three observers. In addition, the automatically segmented tumor volumes from seven separate tumor data sets were quantitatively compared with histology-derived tumor volumes. Results: The highest similarity index between manual and automatic segmentation (SI\u3csub\u3emanual,automatic\u3c/sub\u3e = 0.82 ± 0.06) was observed for k-means clustering with feature vector {T\u3csub\u3e2\u3c/sub\u3e, ADC} and four clusters. A strong linear correlation between automatically and manually segmented tumor volumes (R\u3csup\u3e2\u3c/sup\u3e = 0.99) was observed for this segmentation method. Automatically segmented tumor volumes also correlated strongly with histology-derived tumor volumes (R\u3csup\u3e2\u3c/sup\u3e = 0.96). Conclusion: Automatic segmentation of mouse subcutaneous tumors can be achieved on the basis of endogenous MR contrast only.\u3c/p\u3

MRI methods for the evaluation of high intensity focused ultrasound tumor treatment:current status and future needs

\u3cp\u3eThermal ablation with high intensity focused ultrasound (HIFU) is an emerging noninvasive technique for the treatment of solid tumors. HIFU treatment of malignant tumors requires accurate treatment planning, monitoring and evaluation, which can be facilitated by performing the procedure in an MR-guided HIFU system. The MR-based evaluation of HIFU treatment is most often restricted to contrast-enhanced T\u3csub\u3e1\u3c/sub\u3e-weighted imaging, while it has been shown that the non-perfused volume may not reflect the extent of nonviable tumor tissue after HIFU treatment. There are multiple studies in which more advanced MRI methods were assessed for their suitability for the evaluation of HIFU treatment. While several of these methods seem promising regarding their sensitivity to HIFU-induced tissue changes, there is still ample room for improvement of MRI protocols for HIFU treatment evaluation. In this review article, we describe the major acute and delayed effects of HIFU treatment. For each effect, the MRI methods that have been - or could be - used to detect the associated tissue changes are described. In addition, the potential value of multiparametric MRI for the evaluation of HIFU treatment is discussed. The review ends with a discussion on future directions for the MRI-based evaluation of HIFU treatment.\u3c/p\u3

Dual-targeting of alpha(v)beta(3) and galectin-1 improves the specificity of paramagnetic/fluorescent liposomes to tumor endothelium in vivo

Molecular imaging of angiogenesis requires a highly specific and efficient contrast agent for targeting activated endothelium. We have previously demonstrated that paramagnetic and fluorescent liposomes functionalized with two angiogenesis-specific ligands, the galectin-1-specific anginex (Anx) and the a vß 3integrin-specific RGD, produce synergistic targeting effect in vitro. In the current study, we applied Anx and RGD dual-conjugated liposomes (Anx/RGD-L) for angiogenesis-specific MRI in vivo, focusing on the specificity and efficacy of liposome association with tumor endothelium. The targeting properties, clearance kinetics and biodistribution of Anx/RGD-L were investigated in B16F10 melanoma-bearing mice, and compared to liposomes functionalized with either Anx (Anx-L) or RGD (RGD-L). The contrast enhancement produced by dual- and single-targeted nanoparticles in the tumor was measured using in vivo T 1-weighted MRI, complemented by ex vivo immunohistochemical evaluation of tumor tissues. Blood clearance kinetics of Anx/RGD-L was three-fold more rapid than for RGD-L, but comparable to Anx-L. Both dual- and single-targeted liposomes produced similar changes in MRI contrast parameters in tumors with high inter-tumor variability (¿R 1 = 0.04 ± 0.03 s -1, 24 h post-contrast). Importantly, however, the specificity of Anx/RGD-L association with tumor endothelium of 53 ± 6%, assessed by fluorescence microscopy, was significantly higher compared to 43 ± 9% (P = 0.043) and 28 ± 8% (P = 0.0001) of Anx-L and RGD-L, respectively. In contrast, long-circulating RGD-L were on average 16% more efficient in targeting tumor endothelium compared to Anx/RGD-L. Significant differences were also found in the biodistribution of investigated contrast agents. In conclusion, synergistic targeting of a vß 3 and galectin-1 improved the specificity of the association of the liposomal contrast agent to tumor endothelium in vivo, providing therefore a more reliable MRI readout of the angiogenic activity. © 2011 Elsevier B.V. All rights reserved