Magnetic resonance tumor regression grade (MR-TRG) to assess pathological complete response following neoadjuvant radiochemotherapy in locally advanced rectal cancer

This study aims to evaluate the feasibility of a magnetic resonance (MR) automatic method for quantitative assessment of the percentage of fibrosis developed within locally advanced rectal cancers (LARC) after neoadjuvant radiochemotherapy (RCT). A total of 65 patients were enrolled in the study and MR studies were performed on 3.0 Tesla scanner; patients were followed-up for 30 months. The percentage of fibrosis was quantified on T2-weighted images, using automatic K-Means clustering algorithm. According to the percentage of fibrosis, an optimal cut-off point for separating patients into favorable and unfavorable pathologic response groups was identified by ROC analysis and tumor regression grade (MR-TRG) classes were determined and compared to histopathologic TRG. An optimal cut-off point of 81% of fibrosis was identified to differentiate between favorable and unfavorable pathologic response groups resulting in a sensitivity of 78.26% and a specificity of 97.62% for the identification of complete responders (CRs). Interobserver agreement was good (0.85). The agreement between P-TRG and MR-TRG was excellent (0.923). Significant differences in terms of overall survival (OS) and disease free survival (DFS) were found between favorable and unfavorable pathologic response groups. The automatic quantification of fibrosis determined by MR is feasible and reproducible

Quantitative assessment of diffuse myocardial fibrosis in II-type diabetes mellitus patients using T1 mapping technique: preliminary data

In diabetic cardiomyopathy (DCM), ventricular remodelling consists in a progressive impairment of myocardial contraction (evolving from diastolic to combined diastolic-systolic dysfunction) occurring regardless of ischemic heart disease, hypertension or other macrovascular complications, which ultimately leads to heart failure. Early stages of DCM are asymptomatic and characterised by initial contractile disfunction and various degree of myocardial fibrosis, that may not be recognised by traditional cardiology tests. Our purpose was to detect myocardial fibrotic infiltration in DM-II patients by using T1-mapping technique with extracellular volume fraction (ECV) measurement

Magnetic resonance of rectal cancer response to therapy: an Image quality comparison between 3.0 and 1.5 Tesla

Purpose. To evaluate signal intensity (SI) differences between 3.0 T and 1.5 T on T2-weighted (T2w), diffusion-weighted imaging (DWI), and apparent diffusion coefficient (ADC) in rectal cancer pre-, during, and postneoadjuvant chemoradiotherapy (CRT). Materials and Methods. 22 patients with locally advanced rectal cancer were prospectively enrolled. All patients underwent T2w, DWI, and ADC pre-, during, and post-CRT on both 3.0 T MRI and 1.5 T MRI. A radiologist drew regions of interest (ROIs) of the tumor and obturator internus muscle on the selected slice to evaluate SI and relative SI (rSI). Additionally, a subanalysis evaluating the SI before and after-CRT (ΔSI pre-post) in complete responder patients (CR) and nonresponder patients (NR) on T2w, DWI, and ADC was performed. Results. Significant differences were observed for T2w and DWI on 3.0 T MRI compared to 1.5 T MRI pre-, during, and post-CRT (all P<0.001), whereas no significant differences were reported for ADC among all controls (all P>0.05). rSI showed no significant differences in all the examinations for all sequences (all P>0.05). ΔSI showed significant differences between 3.0 T and 1.5 T MRI for DWI-ΔSI in CR and NR (188.39±166.90 vs. 30.45±21.73 and 169.70±121.87 vs. 22.00±31.29, respectively, all P 0.02) and ADC-ΔSI for CR (-0.58±0.27 vs. -0.21±0.24P value 0.02), while no significant differences were observed for ADC-ΔSI in NR and both CR and NR for T2w-ΔSI. Conclusion. T2w-SI and DWI-SI showed significant differences for 3.0 T compared to 1.5 T in all three controls, while ADCSI showed no significant differences in all three controls on both field strengths. rSI was comparable for 3.0 T and 1.5 T MRI in rectal cancer patients; therefore, rectal cancer patients can be assessed both at 3.0 T MRI and 1.5 T MRI. However, a significant DWI-ΔSI and ADC-ΔSI on 3.0 T in CR might be interpreted as a better visual assessment in discriminating response to therapy compared to 1.5 T. Further investigations should be performed to confirm future possible clinical application

Characterization of Ku702–NLS as Bipartite Nuclear Localization Sequence for Non-Viral Gene Delivery

Several barriers have to be overcome in order to achieve gene expression in target cells, e.g. cellular uptake, endosomal release and translocation to the nucleus. Nuclear localization sequences (NLS) enhance gene delivery by increasing the uptake of plasmid DNA (pDNA) to the nucleus. So far, only monopartite NLS were analysed for non-viral gene delivery. In this study, we examined the characteristics of a novel bipartite NLS like construct, namely NLS Ku70. We synthesized a dimeric structure of a modified NLS from the Ku70 protein (Ku702-NLS), a nuclear transport active mutant of Ku702-NLS (s1Ku702-NLS) and a nuclear transport deficient mutant of Ku702-NLS (s2Ku702). We examined the transfection efficiency of binary Ku702-NLS/DNA and ternary Ku702-NLS/PEI/DNA gene vector complexes in vitro by using standard transfection protocols as well as the magnetofection method. The application of Ku702-NLS and s1Ku702-NLS increased gene transfer efficiency in vitro and in vivo. This study shows for the first time that the use of bipartite NLS compounds alone or in combination with cationic polymers is a promising strategy to enhance the efficiency of non-viral gene transfer

Comparison of Gene-Transfer Efficiency in Human Embryonic Stem Cells

Technologies designed to allow manipulation and modification of human embryonic stem (hES) cells are numerous and vary in the complexity of their methods, efficiency, reliability, and safety. The most commonly studied and practiced of these methods include electroporation, lipofection, nucleofection, and lentiviral transduction. However, at present, it is unclear which protocol offers the most efficient and reliable method of gene transfer to hES cells. In this study, a bi-fusion construct with ubiquitin promoter driving enhanced green fluorescent protein reporter and the firefly luciferase (pUb-eGFP-Fluc) along with neomycin selection marker was used for in vitro and in vivo studies. In vitro studies examined the transfection efficiency and viability of each technique using two hES cell lines (male H1 and female H9 cells). Lentiviral transduction demonstrated the highest efficiency (H1: 25.3 ± 4.8%; H9: 22.4 ± 6.5%) with >95% cell viability. Nucleofection demonstrated transfection efficiency of 16.1 ± 3.6% (H1) and 5.8 ± 3.2% (H9). However, minimal transfection efficiency was observed with electroporation (2.1 ± 0.4% (H1) and 1.9 ± 0.3% (H9)) and lipofection (1.5 ± 0.5% (H1) and 1.3 ± 0.2% (H9); P < 0.05 vs. lentiviral transduction). Electroporation also demonstrated the highest cell death (62 ± 11% (H1) and 42 ± 10% (H9)) followed by nucleofection (25 ± 9% (H1) and 30 ± 15 (H9)). Importantly, lentiviral transduction generated a greater number of hES cell lines stably expressing the double-fusion reporter gene (hES-DF) compared to other transfection techniques. Finally, following subcutaneous transplantation into immunodeficient nude mice, the hES-eGFP-Fluc cells showed robust proliferation as determined by longitudinal bioluminescence imaging. In summary, this study demonstrates that lentiviral transduction and nucleofection are efficient, simple, and safe techniques for reliable gene transfer in hES cells. The double-fusion construct provides an attractive approach for generating stable hES cell lines and monitoring engraftment and proliferation in vitro and in vivo

Imaging edema in immune checkpoint inhibitor myocarditis: a moving target

We greatly enjoyed reading the recent paper by Thavendiranathan et al. reporting on cardiac magnetic resonance findings in patients with immune checkpoint inhibitor-myocarditis (ICI-M), and we would be glad to share few considerations. Extensive myocardial edema can be found in the acute phase of ICI-M, leading to the concomitant increase of native T1 and T2