1) and was characterized by 1H NMR (Fig. 2012;57:549-555, 9. Molecular architecture of intermediate filaments. This site needs JavaScript to work properly. [Abstract] eCollection 2018 Jun. Lymphocyte activation induces rapid changes in nuclear and cytoplasmic glycoproteins. 2000 Nov;19(11):647-77. doi: 10.1089/10445490050199054. Epub 2011 Mar 21. Li F, Song Z, Li Q. et al. Next, we labeled untargeted PEI-1800 with 99mTc using the same method as 99mTc-GlcNAc-PEI for comparison. We have demonstrated that vimentin, although virtually absent in normal human muscle fibers, is expressed in regenerating fibers in different neuromuscular disorders. var thismailbody2 = ""; There were several other radiotracers reported to image liver fibrosis in mouse or rat models. Scale bar: 200 μm. 2012;33:2154-2164, 17. 2008;134(6):1670-1681, 8. 2008;134:1655-1669, 2. Puche JE, Saiman Y, Friedman SL. In humans, desmin related myopathies might be caused by mutations in the corresponding desmin gene or in proteins with which desmin var thismailbody11 = ""; Sci Rep. 2016;6:28812, 26. This panel serves to rule out rhabdomyosarcoma, leiomyosarcoma, sarcomatoid carcinoma (or other cytokeratin positive sarcoma), and spindle cell melanoma respectively. Zhang D, Guo Z, Zhang P. et al. The purpose of this study was to assess the potential usefulness of 99mTc-GlcNAc-PEI for SPECT/CT monitoring of the progression and prognosis of liver fibrosis in a mouse model. (1993).  |  3A. Extracellular matrix (ECM) accumulation in liver fibrosis is caused by the activation of hepatic stellate cells (HSCs). Nestin, Vimentin, and Desmin Expression after In Situ Necrosis The results after in situ necrosis were overall similar, but the regeneration seemed to advance slightly ahead of that seen in the shearing-type injury. Helps differentiate smooth muscle tumors (desmin+) from GIST (c-kit+, almost always desmin-) In serous effusion, more common in reactive mesothelial cells (84%) than mesothelioma (8%) or carcinoma (2%, Am J Surg Pathol 2001;25:1405)  |  At the transition from linear to nonlinear viscoelastic response, only desmin shows characteristics of nonaffine network deformation. Results: 99mTc-GlcNAc-PEI is a hydrophilic compound with high radiochemical purity (>98%) and good stability. Histologic analysis of the Sirius Red staining results demonstrated that the collagen fiber was reduced evidently in fibrotic mice after clodronate liposome treatment (Fig. S2A). After the standard curve of GlcNAc was plotted (Fig. Compared with control, clodronate liposome treatment reduced the LUV of 99mTc-GlcNAc-PEI (4.62 ± 0.352 vs 2.133 ± 0.414%/cc; P ˂ 0.05). Differences were considered significant when P values were less than 0.05. These results suggested that 99mTc-GlcNAc-PEI SPECT/CT imaging could be used as a potent method to monitor fibrosis progression and evaluate the effect of anti-fibrotic drugs. (B) Hepatic uptake of control and fibrotic mice by drawing the ROI of the whole liver. Meanwhile, vimentin expression was observed in the tensor veli palatine muscle and surrounding tissue at ED 12.5, and this level of expression did not change up to ED 15.5. There were strong correlations between the hydroxyproline level and Sirius Red quantification (R = 0.88, Fig. When applying whole cell extracts of cell lines expressing vimentin plus glial fibrillary acidic protein (GFAP), and vimentin plus desmin, respectively, the antibody labels specifically the 57 kDa vimentin band. 4A), respectively, indicating non-specific uptake. Gastroenterology. A similar protein, vimentin, is present in higher amounts during embryogenesis while desmin is present in higher amounts after differentiation. 18F-FDG PET/CT imaging and 99mTc-GSA SPECT/CT imaging confirmed the pathologic process of fibrosis in the CCl4 mice (Fig. Immunofluorescent assay showed strong colocalization of anti-desmin (Fig. In aortic tissue some smooth muscle cells contain both types of intermediate filament proteins, vimentin and desmin. The RCP of 99mTc-GlcNAc-PEI was still greater than 92% when incubated in PBS or serum for 4 h, as determined by TLC (shown in Fig. Gene delivery system based on highly specific recognition of surface-vimentin with N-acetylglucosamine immobilized polyethylenimine. Characterization of a CCl4-induced mouse model of liver fibrosis. (A) SPECT/CT imaging of CCl4-induced fibrotic mice with 99mTc-GlcNAc-PEI. Pharmacological JNK2 inhibition produced results qualitatively similar to Blocking studies of 99mTc-GlcNAc-PEI in fibrotic mice. Control group was administered with same doses of olive oil (n = 5 for each group). The necrosis forging ahead from the transection site within the basal lamina had been halted by contraction bands. Desmin is stiffer than vimentin (l p desmin ≈ 900 nm; l p vimentin ≈ 400 nm under LS conditions), but both IFs are much more flexible than microfilaments. 1996;24:301-307, 4. Localization and distribution of desmin and vimentin have been studied in different neuromuscular disorders using monoclonal antibodies. By contrast, co-injection of an excess dose of GlcNAc-PEI had no effect on the hepatic uptake of 99mTc-GlcNAc-PEI in the liver of normal mice. Biomaterials. Strong correlations were observed between hepatic uptake measured by SPECT/CT and Sirius Red quantification (R = 0.92, Fig. document.write(thismailbody4 + thismailbody3+thismailbody2) COVID-19 is an emerging, rapidly evolving situation. To gain insight into the role of IF proteins in podocytes, we investigated the expression of nestin, vimentin, and desmin in puromycin aminonucleoside (PAN) nephrosis. Liquid chromatographic determination of hydroxyproline in tissue samples. The percentage of the injected dose per gram (%ID/g) of organs and tissues was calculated. *P <0.05 and **P <0.01. The data were analyzed using GraphPad Prism 5. The time-activity curves and representative images derived from the 50-min dynamic SPECT imaging (Fig. … Liver fibrosis is the result of repeated damage to the liver in conjunction with sustained accumulation of extracellular matrix (ECM) proteins1. In the SPECT imaging study, the spleen uptake values in the control and blocking groups were 28.46 ± 2.54%/cc vs 25.40 ± 1.58%/cc (P = 0.15, Fig. The animals were anesthetized by 2% isoflurane during SPECT imaging. The reaction mixture was then allowed to incubate at room temperature for 30 min to obtain the resulting radiotracer 99mTc-GlcNAc-PEI. Fluorescent cell uptake of GlcNAc-PEI-Cy5.5 was decreased obviously when blocked with excess GlcNAc-PEI in HSCs isolated from the liver of fibrotic mice (Fig. Compared with these previously reported radiotracers, we developed a new agent of 99mTc-GlcNAc-PEI by specifically targeting desmin and vimentin to activate HSCs, reflecting liver fibrosis directly and could be used to detect liver fibrosis as early as 4 weeks after CCl4 treatment. The ability of SP-A to interact with desmin and vimentin, and to prevent polymerization of desmin monomers, shed The ability of SP-A to interact with desmin and vimentin, and to prevent polymerization of desmin monomers, shed light on unexpected and wider biological roles of this collectin. The expression pattern of desmin in smooth muscle is much more heterogenous. Both changes in radioactivity accumulation in the liver do not reflect the degree of fibrosis directly and therefore cannot be used for liver fibrosis diagnosis. Next, 185~370 MBq of freshly Na99mTcO4 obtained from Zhongshan Hospital affiliated with Xiamen University was added to the mixture. Peroxisome proliferator-activated receptors and hepatic stellate cell activation. In the absence of vimentin networks (MCF-7 cells), the initiation of desmin filament formation seems to start on the preexisting keratin filaments. S6) staining (FITC) with Cy5.5-labeled GlcNAc-PEI (GlcNAc-PEI-Cy5.5), confirming that GlcNAc-PEI could target desmin and vimentin on the surface of activated HSCs. Kim SJ, Ise H, Kim E. et al. 2011;32:3471-3480, 31. The Glu108 position is fully conserved among desmin, vimentin, neurofilament L protein, cytokeratins 8 and 18, and nuclear lamins A and B1. Proc Natl Acad Sci U S A. Tolstonog GV, Wang X, Shoeman R, Traub P. Tolstonog GV, et al. Unlike 99mTc-GlcNAc-PEI, the increased uptake of 18F-FDG (Fig. 1999;29:520-527, 16. J Hepatol. World J Gastroenterol. There is an urgent need to develop non-invasive ways to assess liver fibrosis in chronic liver disease and monitor fibrosis progression and the treatment response. (B) Hepatic uptake of 99mTc-GlcNAc-PEI by drawing the ROI of the whole liver. S7). *P <0.05 and **P <0.01. After imaging, the livers of the control and fibrotic mice were frozen in CRYO GLUE tissue embedding medium (SLEE medical GmbH, Mainz, GER) and were stored in a -80 °C freezer. 99mTc-GlcNAc-PEI SPECT/CT static imaging could detect the difference between these two groups of mice in the evaluation of liver fibrosis. Komura K, Ise H, Akaike T. Dynamic behaviors of vimentin induced by interaction with GlcNAc molecules. All cats were European short hairs. Static SPECT/CT imaging of fibrotic (CCl4-4 W and CCl4-8 W) and control mice (n = 4 for each group) were performed at 30 min after tail-vein injection of 18.5 MBq of 99mTc-GlcNAc-PEI. creased,26 whereas levels of desmin and vimentin are in- creased.22,26 We have recently reported that activated stellate cells also produce nestin, another IF protein.27 Human quies- cent stellate cells contain vimentin; a subpopulation of acti-vated stellate cells also express desmin.28 The biological sig- nificance of the simultaneous occurrence of different IF (A) 99mTc-GlcNAc-PEI SPECT/CT imaging of fibrotic mice with clodronate liposome treatment. た。しかしdesminは顎舌骨筋において,メッケル軟骨に近接する筋束に強い発現が観察された。さらに胎生 16日ではvimentinの発現傾向は変わらないものの,desminの発現は顎舌骨筋全体に拡がっていた … We obtained a similar result with autoradiography of the liver tissue of the control and fibrotic mice (Fig. Desmin is a myogenic marker expressed in early development that forms a network of filaments that extends across the myofibril and surrounds Z discs. J Biol Chem. Radioactivity of the cell was measured with a gamma counter (WIZARD 2480, Perkin-Elmer, Waltham, MA, USA). The lack of non-invasive ways to monitor fibrosis progression limited the development of anti-fibrotic drugs20. Blue: DAPI; green: anti-CD68 staining FITC; Red: GlcNAc-PEI-Cy5.5. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics & Center for Molecular Imaging and Translational Medicine, School of Public Health, Xiamen University, Xiamen 361102, China.2. vimentin and desmin. 285 g of iodine. in Fibroblasten, Chondrozyten, Endothelzellen und glatten Muskelzellen. Influence of abstinence, sex and extent of hepatic damage. (F) Immunofluorescence co-localization analysis of desmin and GlcNAc-PEI-Cy5.5 in the liver tissue of CCl4-induced fibrotic mice. However, the high uptake of those radiotracers in the kidney influenced the quality of liver images and was not good for the quantitation of hepatic uptake. Furthermore, desmin and vimentin overexpressions did not alter BSM contractility and MLC 20 phosphorylation in strips isolated from JNK2 knockout mice. 2003;791:427-430. Interactions of vimentin- or desmin-expressing liver cells with N-acetylglucosamine-bearing polymers. Rockey DC, Caldwell SH, Goodman ZD. Mechanical Properties of the Cytoskeleton and Cells. [Abstract] Blocking experiments were performed with the co-injection of an excess dose of GlcNAc-PEI (500 μg). Polasek M, Fuchs BC, Uppal R. et al. 1986;2:33-42, 27. Norlén L, Masich S, Goldie KN, Hoenger A. Exp Cell Res. Necrosis was induced by hot Ringer solution. To confirm the desmin and vimentin specificity of 99m Tc-GlcNAc-PEI in the fibrotic liver, blocking experiments were performed. 4B). Posey D, Blaisdell-Pijuan P, Knoll SK, Saif TA, Ahmed WW. NIH 2010;44:637-645, 13. Subsequently, the specimens were cut into 7-μm slices. 2G). Furthermore, there was a very high binding affinity and lower kidney uptake of 99mTc-GlcNAc-PEI ensuring the high quality of SPECT/CT image which might be beneficial to diagnose early and mild hepatic fibrosis in the clinical setting. Desmin is a normal constituent of skeletal muscle fibers; vimentin is contained in myoblasts and connective tissue cells. The labeling efficiency of 99mTc-GlcNAc-PEI was greater than 95% as determined by TLC. Liver fibrosis gradually progressed with time and gradually developed into progressive hepatic fibrosis after the administration of CCl4 for 8 weeks, as was expected and verified by Sirius Red staining (Fig. Conclusion: 99mTc-GlcNAc-PEI SPECT/CT was useful in assessing liver fibrosis and monitoring the treatment response. Noninvasive SPECT imaging with 99mTc-GlcNAc-PEI was used to assess liver fibrosis in a carbon tetrachloride (CCl4) mouse model. (E) Hydroxyproline level in the liver of CCl4-induced mice after treatment with clodronate liposomes. (C) Biodistribution of 99mTc-GlcNAc-PEI in CCl4-induced fibrotic mice (n = 3). (F) Correlation between the total Ishak score and red area of Sirius Red staining. These results suggest that portal myofibroblasts have a significant role in the process of liver fibrosis in dogs. Quantitative data are presented with error bars as the means ± SD. Desmin knockout mice develop cardiomyopathy, skeletal and smooth muscle defects (5). Arginase 2 deficiency results in spontaneous steatohepatitis: a novel link between innate immune activation and hepatic de novo lipogenesis. Veterinary Pathology 2016 29: 3, 260-262 Download Citation. 2020 Apr 7;142(10):100806. doi: 10.1115/1.4046863. Confocal immunofluorescent analysis of choroid plexus in mouse brain using Desmin (D93F5) XP ® Rabbit mAb (green).After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900, the tissue was then labeled using Ras (E4K9L) Rabbit mAb (Alexa Fluor ® 647 Conjugate) #37182 (red). S1D), the mass percent of sugar in GlcNAc-PEI was calculated as 42.3%. Nanotheranostics 2021; 5(1): 113-124. 1999 Mar 12;286(5):1403-20. doi: 10.1006/jmbi.1999.2528. The LUV of the CCl4 groups was compared with that of the olive oil group. 細胞はKeratin,Vimentinに 陽性を示し,ま た,卵 巣癌の腫瘍マーカーとして知られるCA 125に も高率に陽性を示した.そ の他,Desmin,EMA,S-100蛋 自,腫 瘍マーカーであるCEA,CA 19-9, AFPに ついては,中 皮細胞 … Extracellular matrix (ECM) accumulation in liver fibrosis is caused by the activation of hepatic stellate cells (HSCs). 1. 99mTc-GlcNAc-PEI SPECT/CT imaging of liver fibrosis mice with clodronate liposome treatment. GlcNAc-PEI was successfully radiolabeled with 99mTc using the multi-amino structure of PEI at room temperature with a high radiolabeling efficiency and radiochemical stability. Vimentin is the main intermediate filament protein in mesenchymal cells. Diagnosis and quantitation of fibrosis. (C) Hydroxyproline level in the liver of CCl4-induced mice was calculated by HPLC analysis. Molecular imaging of hepatic stellate cell activity by visualization of hepatic integrin alphavbeta3 expression with SPECT in rat. Es handelt sich um ein Intermediärfilament vom Typ 3 - eine Gruppe, zu der auch das Desmin zählt. Popov Y, Schuppan D. Targeting liver fibrosis: Strategies for development and validation of antifibrotic therapies. In addition at ED 12.5, desmin was observed to accumulate in the area of the myotendinous junction, and this accumulation remained unchanged up to ED 15.5. 2012;22:1741-1759, 15. Golde T , Huster C , Glaser M , Händler T , Herrmann H , Käs JA , Schnauß J . Bovine aorta contains, besides cells in which vimentin and desmin seem to co-exist, distinct bundles of smooth muscle cells, located in outer regions of the tunica media, et al. Desmin may be used as part of a very useful panel for initial work up of a high grade spindle cell neoplasm: desmin, Smooth Muscle Actin (SMA), Pancytokeratin , and S100 Protein. Simultaneous SPECT imaging of multi-targets to assist in identifying hepatic lesions. Combined SPECT/CT imaging provides accurate anatomic data on CT, as well as functional information on SPECT images, facilitating the accurate staging of liver fibrosis19. Hepatology. Twenty-four hours later, the cells were washed twice with binding buffer (phosphate buffer solution with 0.5% BSA) and for 1 h at 37 °C with 1.85 kBq of 99mTc-GlcNAc-PEI. SMA, vimentin and desmin-positive cells in the portal triad was in correlation with the degree of fibrosis, which cannot be said for HSC. S4) showed that the hepatic uptake of fibrotic mice was higher than that of control mice and peaked within 25~30 min after injection. Desmin is a normal constituent of skeletal muscle fibers; vimentin is contained in myoblasts and connective tissue cells. Friedrich-Rust M, Ong MF, Martens S. et al. thismailbody3 = thismailbody3+"2e;"; It is believed to be involved with the intracellular transport of proteins between the nucleus and plasma membrane. Here, we developed a method based on SPECT imaging with 99mTc-GlcNAc-PEI to monitor the therapeutic efficacy of liver fibrosis. S9), a finding that was different from that with 99mTc-GlcNAc-PEI. Therefore, 99mTc-GlcNAc-PEI SPECT imaging could be a useful method in monitoring the response to treatment in live fibrosis. Arthur MJP. Scale bar: 50 μm.