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Showing papers on "Smoothelin published in 2014"


Journal ArticleDOI
TL;DR: The structure of the nondilated and dilated aortic wall in bicuspidy and tricuspid are intrinsically different, with the latter having more aspects of aging, including a defective smooth muscle cell differentiation possibly linked to lowered lamin A/C expression.

66 citations


Journal ArticleDOI
TL;DR: By using UV-assisted capillary force lithography (CFL) to engineer a polyurethane substratum of defined nanotopography and stiffness, this study demonstrates that CFL can facilitate the differentiation of cultured vSMCs, reduce their inflammatory signature, and potentially promote the optimal functioning of the vS MC contractile and cytoskeletal machinery.
Abstract: Vascular smooth muscle cells (vSMCs) retain the ability to undergo modulation in their phenotypic continuum, ranging from a mature contractile state to a proliferative, secretory state. vSMC differentiation is modulated by a complex array of microenvironmental cues, which include the biochemical milieu of the cells and the architecture and stiffness of the extracellular matrix. In this study, we demonstrate that by using UV-assisted capillary force lithography (CFL) to engineer a polyurethane substratum of defined nanotopography and stiffness, we can facilitate the differentiation of cultured vSMCs, reduce their inflammatory signature, and potentially promote the optimal functioning of the vSMC contractile and cytoskeletal machinery. Specifically, we found that the combination of medial tissue-like stiffness (11 MPa) and anisotropic nanotopography (ridge width_groove width_ridge height of 800_800_600 nm) resulted in significant upregulation of calponin, desmin, and smoothelin, in addition to the downregul...

49 citations


Journal ArticleDOI
TL;DR: The smooth muscle content (as a proportion of the vascular cross-sectional area) of endometrial blood vessels remained unchanged during the normal menstrual cycle and in menorrhagia; however, expression of the VSMC differentiation markers, smoothelin and calponin, was dysregulated in endometrian blood vessels in samples from women with menor rhagia compared with controls.
Abstract: analysed in relation to total number of blood vessels by double immunostaining for endothelial cell markers. main results and the role of chance: Study of VSMC differentiation markers revealed decreased expression of calponin both in aSMA + vessels (P ¼ 0.008) and in relation to total number of vessels (P ¼ 0.001) in late secretory phase endometrium in menorrhagia compared with controls. Smoothelin expression inaSMA + vessels was increased (P ¼ 0.03) in menorrhagia, although this was not significant in relation to the total number of vessels. In normal endometrium, the proportion of blood vessels expressing aSMA increased from 63% in proliferative endometrium to 81% in the late secretory phase (P ¼ 0.002). The overall arterial muscle content did not differ between control and menorrhagia at any phase of the menstrual cycle, occupying 78–81% of gross vascular cross-sectional area during the different menstrual cycle phases. limitations, reasons forcaution: This study included both straight and spiral arterioles and analysed only stratum functionalis. The VSMC differentiation with respect to aSMA expression is an observational study and the data are presented as presence or absence of the differentiation markers in each field of view, corresponding with the vascular cross sections included in the study of vascular muscle content. wider implications of the findings: Smoothelin and calponin have been widely implicated as important regulators of vascular tone, vascular contractility and rate of blood flow. Our results have uncovered a disparate pattern of calponin expression, potentially indicating a dysfunctional contraction mechanism in the endometrial blood vessels in menorrhagia, thus implicating calponin as a potential therapeutic target.

25 citations


Journal ArticleDOI
TL;DR: Preliminary results suggest global SMTNL1 deletion is associated with greater myogenic reactivity of cerebral arterioles, although the precise mechanism accounting for this finding remains to be defined.
Abstract: Vascular smooth muscle contraction and the myogenic response regulate blood flow in the resistance vascular and contribute to systemic blood pressure. Three pathways are currently known to contribute to the development of the myogenic response: (i) Ca(2+) -dependent phosphorylation of LC20; (ii) Ca(2+) sensitization through inhibition of myosin phosphatase; and (iii) cortical actin polymerization. A number of regulatory smooth muscle proteins are integrated with these pathways to fine tune the response and facilitate adaptations to vascular (patho)physiologies. Of particular interest is the SMTN family of proteins, consisting of SMTN-A, SMTN-B, and the SMTN-like protein, SMTNL1. The SMTN-B and SMTNL1 proteins are both implicated in regulating smooth muscle contractility and contributing to vascular adaptations associated with hypertension, pregnancy, and exercise training. In the case of SMTNL1, the protein plays multiple roles in regulating contraction through functional interactions with contractile regulators as well as transcriptional control of the contractile phenotype and Ca(2+) -sensitizing capacity. For the first time, preliminary results suggest SMTNL1 is involved in the myogenic response of the cerebral resistance vasculature. In this regard, global SMTNL1 deletion is associated with greater myogenic reactivity of cerebral arterioles, although the precise mechanism accounting for this finding remains to be defined.

24 citations


Journal ArticleDOI
TL;DR: Focal cavity lining of the cavity was observed in almost all cases, and the immunophenotype was identical to that of GC wall cells, suggesting features of early muscle differentiation, without ruling out a myofibroblastic origin.
Abstract: 3378047The aim of this study was to characterize the morphology and immunophenotype of ganglion cysts (GCs) and explore their histogenetic origin. Material and Methods: A cross-sectional morphological and immunohistochemical study of 354 GCs used the following antibody panel: vimentin, specific actin, s-actin, smooth-muscle actin, smoothelin, h-caldesmon, s-catenin, desmin, calponin, podoplanin, keratins 5/6, E-cadherin, cyclooxygenase 2 (COX-2), lysozyme, CD10, CD31, CD33, CD34, CD68, Ki-67, and PCNA. Double-blind semi-quantitative analyses were conducted to evaluate the immunopositivity on a 4-point scale. Samples from 10 synovial membranes and 10 scapholunate ligaments were compared. GCs showed a hyalinized wall with mesenchymal spindle cells and were intensely positive for vimentin, actins, h-caldesmon, calponin in all cases and for podoplanin in 53% of cases, suggesting features of early muscle differentiation, without ruling out a myofibroblastic origin. Focal cavity lining of non-synovial flat or raised cells (CD34/CD31/CD10/E-cadherin-negative and podoplanin-positive in 34% of cases) was detected in 93% of cases, showing differential expression with synovial membrane and scapholunate ligament cells. Nuclear positivity for proliferative markers was observed in GC wall cells (258.1±255; 1019.3±316 positive cells/mm2, Ki-67 and PCNA, respectively) but positivity for these markers was significantly lower (p<0.001 Mann Whitney U-test) in scapholunate ligament samples. Conclusion: In this first immunohistochemical study of GCs, focal cellular lining of the cavity was observed in almost all cases, and the immunophenotype was identical to that of GC wall cells. These cells are immunohistochemically different from synoviocytes and scapholunate ligament cells and show characteristics of myofibroblasts or mesenchymal cells undergoing early muscle differentiation.

10 citations


Journal Article
TL;DR: Compared to SMA, caldesmon and smoothelin are more specific stains that allow better delineation of the muscularis propria from the desmoplastic stromal reaction which provides a critical aide for proper staging of colonic adenocarcinoma and subsequent patient care.
Abstract: An accurate distinction between deep muscularis propria invasion versus subserosal invasion by colonic adenocarcinoma is essential for the accurate staging of cancer and subsequent optimal patient management. However, problems may arise in pathologic staging when extensive desmoplasia blurs the junction between deep muscularis propria and subserosal fibroadipose tissue. To address this issue, forty-three (43) cases of colonic adenocarcinoma resections from 2007-2009 at The Methodist Hospital in Houston, TX were reviewed. These cases were selected to address possible challenges in differentiating deep muscularis propria invasion from superficial subserosal invasion based on HE 12 pT2 tumors; and 31 pT3 tumors. All 51 (100%) had diffuse, strong (3+) immunoreactivity for caldesmon and smoothelin in the muscularis propria with a granular cytoplasmic staining pattern. However, the desmoplastic areas of these tumors, composed of spindled fibroblasts and myofibroblasts, showed negative immunostaining for caldesmon and smoothelin (0/35). SMA strongly stained the muscularis propria and weakly (1+) or moderately (2+) stained the spindled fibroblasts in the desmoplastic areas (the latter presumably because of myofibroblastic differentiation). Compared to SMA, caldesmon and smoothelin are more specific stains that allow better delineation of the muscularis propria from the desmoplastic stromal reaction which provides a critical aide for proper staging of colonic adenocarcinoma and subsequent patient care.

10 citations


Journal ArticleDOI
TL;DR: N-cadherin can play a role as a discriminatory marker for interstitial cells in the human bladder, as the interstitial compartment of the human urinary bladder houses a population of cells from mesenchymal origin, immunopositive for N-c cadherin, vimentin, and C-kit.
Abstract: Aims. Interstitial cells, also called myofibroblasts, most probably play a major role in the pathogenesis of the overactive bladder. However, no specific phenotypic marker has been identified. We investigated whether N-cadherin could play a role as a discriminatory marker for interstitial cells in the human bladder. Methods. Bladder biopsies (n = 16) were collected from macroscopically nonpathological locations during cystectomy which was performed because of bladder cancer. Tissue was analyzed for expression of N-cadherin. N-cadherin+ cells were phenotyped using antibodies against PGP9.5, smoothelin, vimentin, and C-kit. Findings were related to bladder tissue histology and ultrastructure of myofibroblastic cells. Results. N-cadherin+/vimentin+ cells with branched cell bodies were found in the lamina propria and detrusor layer. They were closely associated with neurons and showed no colocalization of PGP9.5 or smoothelin. A second type of N-cadherin+ cells was found at the boundary of detrusor bundles and in the lamina propria. These cells colocalization C-kit. We assumed that N-cadherin+/vimentin+ cells are similar to the ultrastructurally defined myofibroblasts. Conclusions. N-cadherin can play a role as a discriminatory marker for interstitial cells in the human bladder, as the interstitial compartment of the human bladder houses a population of cells from mesenchymal origin, immunopositive for N-cadherin, vimentin, and C-kit.

5 citations


Dissertation
01 Mar 2014
TL;DR: Results show cyclic strain at 5% and 10% levels of amplitude increases the abundance of contractile phenotype markers and how the cellular response to this stimulus is orchestrated via miRNA is highlighted.
Abstract: Vascular smooth muscle cells (VSMCs) are major contributors to regulation of vascular tone through their contractile flexibility. Along with other stimulus, they are exposed to mechanical cyclic strain. It is thought such stimulus can be a major regulator of phenotypic modulation in smooth muscle cells. Differentiated SMCs are characteristically quiescent and adapt elongated spindle morphology. In this state the cell expresses an extensive range of contractile proteins such as SM α-actin, Smoothelin and Calponin. SMCs in the synthetic end of the phenotype spectrum adapt a rhomboid morphology and display increased growth rate and higher migratory activity. Expression of the contractile proteins are significantly decreased and replaced with organelles involved in protein synthesis. Studies have shown that several non coding micro RNA (miRNA), which negatively regulate gene expression, are differentially expressed in altered SMC phenotypes. However, work into the effect of mechanical stimulus such as shear stress or cyclic strain on miRNA expression is scarce with only a handful so far identified. This study hypothesizes that “cyclic mechanical strain modulates vascular smooth muscle phenotype and ultimately function, mediated by miRNA regulation” This hypothesis was tested by exposing primary derived human aortic smooth muscle cells (HAoSMCs) to cyclic strain in an in vitro model. Results show cyclic strain at 5% and 10% levels of amplitude increases the abundance of contractile phenotype markers. A temporal study of the miRNA profile of SMCs exposed to 10% strain revealed a significant change in expression was evident after 24 hours of strain. Novel chemotaxis experiments demonstrated cyclic strain reduced cell migration, the effect of which was most evident at 24 hours where migration of the cells was less than half that of the static culture. To implicate miRNA as key coordinators in the reduced cell migration observed after cyclic strain, siRNA were targeted against an essential miRNA biogenesis molecule, Argonaute2 (Ago2). Cells transfected with siRNA for Ago2 negated the physiological stimulus of strain and had a marked increase in migration even over the static control. These investigastions highlight the physiological importance of cyclic strain on SMC phenotype and function how the cellular response to this stimulus is orchestrated via miRNA.

5 citations


Journal ArticleDOI
TL;DR: Investigation of preliminary data suggesting that loss of the α5 chain of collagen IV (α5(IV), placental acid phosphatase (PLAP) expression and smoothelin expression can be used diagnostically as markers of gastrointestinal SMNs found that α5( IV) immunohistochemistry with the A7 antibody is not adiagnostically useful marker of gastrointestinalSMNs.
Abstract: Aims The histological distinction between gastrointestinal smooth muscle neoplasms (SMNs) and their differential diagnoses, especially gastrointestinal stromal tumours (GISTs), has important clinical management implications. This study aimed to investigate preliminary data suggesting that loss of the α5 chain of collagen IV (α5(IV)), placental acid phosphatase (PLAP) expression and smoothelin expression can be used diagnostically as markers of gastrointestinal SMNs. To aid this investigation, these potential markers were directly compared against caldesmon. Methods 31 SMNs and 111 potential differential diagnoses (16 different neoplasm types) were immunostained for caldesmon, PLAP and smoothelin. Results A pilot study indicated that loss of α5(IV) positivity was neither a specific nor sensitive marker of SMN. Caldesmon, PLAP and smoothelin were expressed by all 31 SMNs though leiomyosarcomas showed some loss of staining proportion and/or intensity. Caldesmon positivity was commonly shown by GISTs, glomus tumours and angiomyolipomas. Cytoplasmic smoothelin positivity was commonly shown by glomus tumours and angiomyolipomas, whereas PLAP positivity was shown by one desmoplastic small round cell tumour studied and only infrequently shown by angiomyolipomas. Nuclear smoothelin positivity was seen among four leiomyosarcomas but also a wide range of non-SMNs. Conclusions α5(IV) immunohistochemistry with the A7 antibody is not a diagnostically useful marker of gastrointestinal SMNs. Both PLAP and smoothelin (cytoplasmic expression only) are as sensitive as but are more specific than caldesmon as such a marker. Further, PLAP and smoothelin immunostainings are technically reliable and can be reproducibly assessed.

5 citations


Journal ArticleDOI
TL;DR: In presence of an atypical imaging of the gallbladder, diagnosis of this group of congenital anomalies should be considered in order to adequately plan surgical intervention if necessary.
Abstract: Gallbladder duplication is a rare congenital anomaly, with an incidence of 1 in 3,800 autopsies. The correct diagnosis and treatment of this type of entity is important in clinical practice, because it may cause some clinical and surgical problems. In this report, we present the clinical case of a 28-year-old female with abdominal pain. Ultrasound of the upper abdomen showed a distended gallbladder with the presence of a septum that could suggest a congenital anomaly of the extrahepatic biliary system. During surgery, a distended and inflamed gallbladder with a lithiasis was found. In addition, a complete septum and double cystic duct were observed. The gross and histopathological evaluation of the surgical specimen allowed us to confirm the diagnosis of a Y- shaped type gallbladder duplication according to Boyden�s classification. In conclusion, in presence of an atypical imaging of the gallbladder, diagnosis of this group of congenital anomalies should be considered in order to adequately plan surgical intervention if necessary

3 citations


Journal ArticleDOI
TL;DR: Exposure to exogenous H2O2 or incubation of dispersed gastric muscle cells with SIN-1 significantly increased the expression of calponin, caldesmon, and smoothelin proteins, suggesting an important role in gastrointestinal motility disorders associated with oxidative stress.
Abstract: Thin filament-associated proteins such as calponin, caldesmon, and smoothelin are believed to regulate acto-myosin interaction and thus, muscle contraction. Oxidative stress has been found to affect the normal contractile behavior of smooth muscle and is involved in the pathogenesis of a number of human diseases such as diabetes mellitus, hypertension, and atherosclerosis. However, very little is known about the effect of oxidative stress on the expression of smooth muscle contractile proteins. The aim of the current study is to investigate the effect of oxidative stress on the expression of thin filament-associated proteins in rat gastric smooth muscle. Single smooth muscle cells of the stomach obtained from Sprague–Dawley rats were used. Muscle cells were treated with hydrogen peroxide (H2O2) (500 μM) for 30 min or the peroxynitrite donor 3-morpholinosydnonimine (SIN-1) (1 mM) for 90 min to induce oxidative stress. Calponin, caldesmon, and smoothelin expressions were measured via specifically designed enzyme-linked immunosorbent assay. We found that exposure to exogenous H2O2 or incubation of dispersed gastric muscle cells with SIN-1 significantly increased the expression of calponin, caldesmon, and smoothelin proteins. In conclusion: oxidative stress increases the expression of thin filament-associated proteins in gastric smooth muscle, suggesting an important role in gastrointestinal motility disorders associated with oxidative stress.

Journal ArticleDOI
TL;DR: The results indicate that there are distinct functional differences between different EGFR mutations, which argue for the development of mutation specific targeted therapies.
Abstract: BACKGROUND: EGFR is frequently mutated in various types of cancer. Although all oncogenic mutations are considered activating, different tumor types have different mutation spectra. It is possible that functional differences underlie this tumor-type specific mutation spectrum. METHODS: We have determined whether specific mutations in EGFR (EGFR, EGFRvIII and EGFR-L858R) have differences in binding partners, differences in downstream pathway activation (gene expression and phosphoproteins), and have functional consequences on cellular growth and migration. RESULTS: Using biotin pulldown and subsequent mass spectrometry we were able to detect mutation specific binding partners for EGFR. Differential binding was confirmed using a proximilty ligation assay and/or Western Blot for the dedicator of cytokinesis 4 (DOCK4), UDP-glucose glycoprotein glucosyltransferase 1 (UGGT1), MYC binding protein 2 (MYCBP2) and Smoothelin (SMTN). We also demonstrate that each mutation induces the expression of a specific set of genes, and that each mutation is associated with specific phosphorylation patterns. Finally, we demonstrate using stably expressing cell lines that EGFRvIII and EGFL858R display reduced growth and migration compared to EGFR wildtype expressing cells. CONCLUSION: Our results indicate that there are distinct functional differences between different EGFR mutations. The functional differences between different mutations argue for the development of mutation specific targeted therapies.

Journal ArticleDOI
TL;DR: Despite the importance of SMTL-B in vascular tone, it is not a target of MMP-2 in LPS-induced hypocontractility and was unaltered in L PS aortas and further unaffected by ONO-4817.
Abstract: Smoothelin-B (SMTL-B) and calponin-1 are important regulators of vascular contraction. SMTL-B contains a calponin-homology domain and is structurally similar to cardiac troponin T. As calponin-1 and troponin T are proteolyzed by intracellular matrix metalloproteinase (MMP)-2 in oxidative stress injury, we hypothesized that SMTL-B is also cleaved by MMP-2 and contributes to lipopolysaccharide (LPS)-induced vascular hypocontractility. Rats received ONO-4817 (an MMP inhibitor) or its vehicle, 2 h prior to being administered lipopolysaccharide (LPS). LPS-induced aorta hypocontractility to potassium chloride or phenylephrine, and reduction of calponin-1 levels, were abolished by ONO-4817 at 6 but not 3 h after LPS. However, the level of SMTL-B was unaltered in LPS aortas and further unaffected by ONO-4817. Despite the importance of SMTL-B in vascular tone, it is not a target of MMP-2 in LPS-induced hypocontractility.