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Objective The aim of this study was to demonstrate the immunohistochemical profile of oral inflammatory myofibroblastic tumors (IMTs) along with morphologic analysis. Study design Three cases diagnosed as oral IMTs were selected to compile an immunohistochemical panel constituted by calponin, caldesmon, Bcl-2, desmin, fibronectin, CD68, Ki-67, S100, anaplastic lymphoma kinase (ALK), α–smooth muscle actin, cytokeratins AE1/AE3, muscle-specific actin, CD34, and vimentin. An oral squamous cell carcinoma with a focal area of desmoplastic stroma was used as control for the stained myofibroblastic cells. Results All oral IMTs were positive for calponin, revealing a strong and diffuse expression in the spindle-shaped cells. The lesions were also positive for vimentin (3/3), fibronectin (3/3), α–smooth muscle actin (3/3), and muscle-specific actin (1/3) and negative for h-caldesmon, Bcl-2, desmin, CD68, Ki-67, S100, ALK, cytokeratins AE1/AE3, and CD34. Conclusions Within the results encountered, the present panel should be of great assistance in the diagnosis of oral IMTs.

We have previously reported that actin undergoes a conformational transition (which we named “maturation”) during polymerization, and that the actin-binding protein, caldesmon (CaD), when added at an early phase of polymerization, interferes with this process (Huang et al. J Biol Chem 2010; 285:71). The pre-transition filament is characterized by relatively low pyrene-fluorescence intensity when pyrene-labeled actin is used as a reporter of subunit assembly into filaments, whereas the mature filament emits a characteristic enhanced fluorescence. Previously reported co-sedimentation experiments suggest that filament formation is not inhibited by the presence of CaD, despite blocking the transition associated with filament maturation. In this study we visualized structural effects of CaD on the assembly of actin filaments by TIRF and electron microscopy. CaD-free actin forms “rough” filaments with irregular edges and indistinct subunit organization during the initial phase (∼20 min under our conditions) of polymerization as reported previously by others (Steinmetz et al. J Cell Biol 1997; 138:559; Galinska-Rakoczy et al. J Mol Biol 2009; 387:869), which most likely correspond to the pre-transition state preceding the maturation step. Later during the polymerization process “mature” filaments exhibit a smoother F-actin appearance with easily detectible double helically arranged actin subunits. While the inclusion of the actin-binding domain of CaD during actin polymerization does not affect the elongation rate, it is associated with a prolonged pre-transition phase, characterized by a delayed alteration (rough to smooth) of the appearance of filaments, consistent with a later onset of the maturation process.

Summary Malignant nasopharygeal neoplasms are relatively rare. Anaplastic nasopharyngeal carcinoma is the most common and the most malignant one. In this paper, we present a case of 56-year old patient with high-grade leiomyosarcoma of nasopharynx. The final diagnosis was achieved by using immunohistochemical methods – SMA (+), Caldesmon (+/−), Desmin (+/−), bcl 2 (+/−), EMA (−), S-100 (−), CD 34 (−), MIB 1 – 70% cells. The patient was successfully treated with radical radiotherapy IMRT technique (total dose 7000 cGy in 7 weeks). Currently, the patient has no symptoms and signs of the disease (oncological follow-up; CT imaging, PET-CT). This case is the first nasopharyngeal leiomyosarcoma reported in Poland, and probably the second one in the world.

The author has recently reported the distribution of the cytoskeleton-associated protein caldesmon in spleen and lymph nodes detected with different antibodies against caldesmon ( J Histochem Cytochem 58:183–193, 2010). Here the author reports the distribution of caldesmon in the CNS and ganglia of the mouse using the same antibodies. Western blot analysis of mouse brain and spinal cord showed the preponderance of l-caldesmon and suggested at least two l-caldesmon isoforms in the brain. Immunostaining revealed the predominant reactivity of smooth muscle cells and cells resembling pericytes of many large and small blood vessels, ependymocytes, and secretory cells of the pineal gland and pituitary gland. Neuronal perikarya and neuropil in general displayed no or weak immunoreactivity, but there was stronger labeling of neuronal perikarya in dorsal root and trigeminal ganglia. In the brain, staining of the neuropil was stronger in the molecular layers of the dentate gyrus and cerebellum. Results show that caldesmon is expressed in many different cell types in the CNS and ganglia, consistent with the notion that l-caldesmon is ubiquitously expressed, but it appears most concentrated in smooth muscle cells, pericytes, epithelial cells, secretory cells, and neuronal perikarya in dorsal root and trigeminal ganglia.

Muscle tissues are generally classified into three types, skeletal, cardiac, and smooth muscle. Smooth muscle cells (SMCs) include all muscle cells other than skeletal and cardiac muscles. There are various types of smooth muscle tissues, and they play important roles in many physiological functions. SMCs are localized in digestive tract (such as esophagus, stomach, ileum, small intestine, large intestine), blood vessels, uterus, airway, and bladder. Response to agonists of these SMCs varied from one muscle type to other type. The main mechanism of activation of contraction is transient increment of cytoplasmic concentration of Ca2+, followed by phosphorylation of myosin light chain. Some actin-binding proteins activate or inhibit the above actomyosin-based contraction. This regulatory mechanism of the contraction is thought to be universal among various SMCs. Since SMCs of these tissues are small and furthermore surrounded by large amount of extra cellular matrix or mucous membrane, we can isolate only a small amount of SMCs purely. For biochemical study of muscle proteins, selected SMCs, such as avian gizzard, bovine stomach, bovine aorta, are usually used for convenient starting materials. Most of genes of major muscle proteins in SMCs are isolated so far. Therefore, these genes can be expressed in bacterial, insect, or cultured animal cells. However, myosin or other muscle proteins have been purified from animal tissues still now, because of complexity of construction of expression vectors or economical reasons of these expression systems. In this report we describe improved and convenient methods to prepare myosin and related regulatory proteins.

Introduction: For the past decades long-term survival of oral squamous cell carcinoma (OSCC) patients remain unchanged which provides a wide platform for research. Caldesmon (CaD), an actin-myosin binding protein, has a contributing role in cytoskeleton modulation and cell motility. The aim of this study is to evaluate the expression of CaD in metastatic and non-metastatic OSCC and to discuss the possible role of CaD in epithelial mesenchymal transition. Materials and Methods: Archival blocks of 25 metastatic and 25 non-metastatic OSCC patients are included where CaD expression is evaluated immunohistochemically. Results: Overall expression and staining intensity of CaD were statistically significant in different grades of metastatic and non-metastatic OSCC. In summary, high expression of CaD is observed in increasing grades of OSCC but the metastatic potential of the tumour doesn’t seem to have any relation with CaD.

Mizuno N, Niitani M, Shiba H, Iwata T, Hayashi I, Kawaguchi H, Kurihara H. Proteome analysis of proteins related to aggressive periodontitis combined with neutrophil chemotaxis dysfunction. J Clin Periodontol 2011; 38: 310–317. doi: 10.1111/j.1600-051X.2010.01693.x. Abstract Aim: Some patients suffering from aggressive periodontitis (Ag-P) also display neutrophil chemotaxis dysfunction. In this study, we attempted to identify the proteins involved in Ag-P associated with neutrophil chemotaxis dysfunction using proteome analysis. Material and Methods: A two-dimensional fluorescence difference gel electrophoresis system was used to detect differences in protein expression between neutrophils from four patients suffering from Ag-P combined with neutrophil chemotaxis dysfunction and those from four controls. Moreover, the mRNA levels of the proteins identified by the above method were examined in neutrophils from four types of subjects using the real-time polymerase chain reaction: twenty patients suffering from Ag-P with or without the dysfunction, 15 patients with chronic periodontitis, and 15 controls. Results: Four proteins, lactoferrin, caldesmon, heat shock protein 70, and stac, displayed a higher protein expression level in the neutrophils from the patients suffering from Ag-P combined with the neutrophil dysfunction than in those from the control group. The caldesmon mRNA levels in the neutrophils from the patients suffering from Ag-P combined with the neutrophil dysfunction were high compared with those in the neutrophils from the patients suffering from the other two types of periodontitis and those from the control group. Conclusion: Caldesmon may be a marker of Ag-P combined with neutrophil chemotaxis dysfunction.

Caldesmon (CaD) is an actin-binding protein that is capable of stabilizing actin filaments. Phosphorylation of CaD is widely accepted in the actin cytoskeletal modeling and promotion of cell migration. In this study, we show that CaD is a downstream phosphorylation substrate of PFTK1, a novel Cdc-2-related ser/thr protein kinase. Our study stemmed from an earlier investigation where we demonstrated that PFTK1 kinase conferred cell migratory advantages in human hepatocellular carcinoma (HCC) cells. Here, we showed that PFTK1-knockdown cells exhibited much reduced CaD phosphorylation and consequently caused dissociation of CaD from the F-actin fibers. The cellular localization of CaD was also altered in the absence of PFTK1. Immunofluorescence analysis revealed that PFTK1-abrogated cells exhibited a diffused and blurred appearance of CaD localization, whereas intact co-localization with F-actins was apparent in PFTK1-expressing cells. Without the binding of CaD to actin, disappearance of actin stress fibers was also evident in PFTK1-abrogated cells. In addition, we found that CaD is also commonly up-regulated in HCC tumors when compared to adjacent non-malignant liver (P = 0.022). Taken together, our results highlight a novel biological cascade that involved the phosphorylation activation of CaD by PFTK1 kinase in promoting formation of actin stress fibers.

We have recently shown that Src induces the formation of podosomes and cell invasion by suppressing endogenous p53, while enhanced p53 strongly represses the Src-induced invasive phenotype. However, the mechanism by which Src and p53 play antagonistic roles in cell invasion is unknown. Here we show that the Stat3 oncogene is a required downstream effector of Src in inducing podosome structures and related invasive phenotypes. Stat3 promotes Src phenotypes through the suppression of p53 and the p53-inducible protein caldesmon, a known podosome antagonist. In contrast, enhanced p53 attenuates Stat3 function and Src-induced podosome formation by upregulating the tumor suppressor PTEN. PTEN, through the inactivation of Src/Stat3 function, also stabilizes the podosome-antagonizing p53/caldesmon axis, thereby further enhancing the anti-invasive potential of the cell. Furthermore, the protein phosphatase activity of PTEN plays a major role in the negative regulation of the Src/Stat3 pathway and represses podosome formation. Our data suggest that cellular invasiveness is dependent on the balance between two opposing forces: the proinvasive oncogenes Src-Stat3 and the anti-invasive tumor suppressors p53-PTEN.

Ianez R F, Buim M E, Coutinho-Camillo C M, Schultz R, Soares F A & Lourenco S V (2010) Histopathology 57, 410–417 Human salivary gland morphogenesis: myoepithelial cell maturation assessed by immunohistochemical markers Aims: Myoepithelial cells are important components of salivary gland structure, aiding the expulsion of saliva from acinar lobules. The aim was to evaluate the expression of smooth muscle actin (SMA), calponin, caldesmon, CD10, CD29, S100 protein, glial fibrillary acidic protein (GFAP) and p63 in myoepithelial cells during salivary gland morphogenesis to understand the maturation process of these cells and their possible use in the diagnosis of salivary gland lesions. Methods and results: Major and minor human salivary glands at various stages of development, derived from fetuses at 8–26 weeks of gestation, were studied immunohistochemically. Fully developed salivary glands were used as controls. The protein p63 was present in all stages of salivary gland morphogenesis from initial bud to terminal bud stage. CD29, S100 and calponin were detected increasingly as salivary gland structure matured and in fully developed salivary gland. Proteins GFAP, CD10 and caldesmon were not observed in myoepithelial cells of salivary glands. Conclusions: The proteins SMA, calponin, CD29, S100 and p63, which are present from the earliest stages of salivary gland maturation, are valuable myoepithelial markers but, although very specific, are not exclusive markers for this cell type.

Bladder regeneration studies have yielded inconclusive results possibly due to the use of unfavorable cells and primitive scaffold design. We hypothesized that human mesenchymal stem cells seeded onto poly(1,8-octanediol-co-citrate) elastomeric thin films would provide a suitable milieu for partial bladder regeneration. POCfs were created by reacting citric acid with 1,8-octanediol and seeded on opposing faces with human MSCs and urothelial cells; normal bladder smooth muscle cells and UCs, or unseeded POCfs. Partial cystectomized nude rats were augmented with the aforementioned POCfs, enveloped with omentum and sacrificed at 4 and 10 weeks. Isolated bladders were subjected to Trichrome and anti-human gamma-tubulin, calponin, caldesmon, smooth muscle gamma-actin, and elastin stainings. Mechanical testing of POCfs revealed a Young's modulus of 138 kPa with elongation 137% its initial length without permanent deformation demonstrating its high uniaxial elastic potential. Trichrome and immunofluorescent staining of MSC/UC POCf augmented bladders exhibited typical bladder architecture with muscle bundle formation and the expression and retention of bladder smooth muscle contractile proteins of human derivation. Quantitative morphometry of MSC/UC samples revealed muscle/collagen ratios approximately 1.75x greater than SMC/UC controls at 10 weeks. Data demonstrate MSC seeded POCfs support partial regeneration of bladder tissue in vivo.

Context: Studies have shown that labor occurs primarily in the night/morning hours. Recently, we identified the human myometrium as a target for melatonin (MEL), the neuroendocrine output signal coding for circadian night. Objective: The purpose of this study was to determine the signaling pathway underlying the effects of MEL on contractility and the contractile machinery in immortalized human myometrial cells. Design: To ascertain the signaling pathway of MEL leading to its effects on myometrial contractility in vitro, we performed gel retraction assays with cells exposed to iodo-MEL (I-MEL) with or without oxytocin and the Rho kinase inhibitor Y27632. I-MEL effects on inositol trisphosphate (IP3)/diacylglycerol (DAG)/protein kinase C (PKC) signaling were also investigated. Additionally, we assayed for caldesmon phosphorylation and ERK1/2 activation. Results: I-MEL was found to activate PKCα via the phospholipase C/IP3/DAG signaling pathway, which was confirmed by PKC enzyme assay. I-MEL did not affect myosin light chain phosphatase activity, and its effects on contractility were insensitive to Rho kinase inhibition. I-MEL did increase phosphorylation of ERK1/2 and caldesmon, which was inhibited by the MAPK kinase inhibitor PD98059 or the PKC inhibitor C1. Conclusions: MEL sensitizes myometrial cells to subsequent procontractile signals in vitro through activation of the phospholipase C/IP3/DAG signaling pathway, resulting in specific activation of PKCα and ERK1/2, thereby phosphorylating caldesmon, which increases actin availability for myosin binding and cross-bridging. In vivo, this sensitization would provide a mechanism for the increased nocturnal uterine contractility and labor that has been observed in late-term human pregnancy.

In this article, we supplement the few published articles by describing the clinical and pathologic features of pleomorphic and dedifferentiated leiomyosarcoma from 41 patients (27 women and 14 men) with an age range of 25 to 75 years (mean, 56.5 years), representing the largest cohort reported to date. The typical leiomyosarcoma component accounted for

The glucocorticoids (GCs), the most downstream effectors of the hypothalamic-pituitary-adrenal (HPA) axis, are the main mediators of stress response. Stress-triggered GCs as well as acute and chronic GC treatment can impair the structural plasticity and function of the brain. The exposure of perinatal animals and humans to excess stress or GCs can affect the brain development, resulting in altered behaviors in the adult offspring of animals and an increased risk of psychiatric disorders in humans. Despite the numerous studies documenting these effects, the underlying mechanism remains unclear. In this commentary we will focus on the effect of excess GCs on cortical development. We have recently showed that excess-GC-dependent retardation of the radial migration of neural progenitor cells (NPCs) is caused by the dysregulation of actin-myosin interaction via upregulation of caldesmon (CaD), an actin-linked regulatory protein. The elucidation of the molecular mechanisms that underlie the detrimental action of GCs on cortical development will expand our understanding of how stress/GCs alter the formation of neural networks and affect behaviors later in life.

The purpose of this study is to characterize the smooth muscle differentiation of purified human muscle-derived cells (hMDCs). The isolation and purification of hMDCs were conducted by modified preplate technique and Dynal CD34 cell selection. Smooth muscle cell differentiation was induced by the use of smooth muscle induction medium (SMIM) and low-serum medium. The gene expressions at the mRNA and protein levels of undifferentiated and differentiated hMDCs were tested by RT-PCR, Western blot and immunofluorescence studies. Western blot and immunofluorescence studies demonstrated the purified hMDCs cultured in SMIM for 4 weeks and expressed significant amount of smooth muscle myosin heavy chain (MHC) and α-smooth muscle actin (ASMA). The cells cultured in low-serum medium for 4 weeks also expressed ASMA, while the control group did not. RT-PCR analysis showed increased gene expression of smooth muscle markers, such as ASMA, Calponin, SM22, Caldesmon, Smoothelin and MHC when purified hMDCs were exposed to SMIM for 2 and 4 weeks when compared to the controls. In conclusion, we confirmed the smooth muscle differentiation capability of purified hMDCs. The gene expression of smooth muscle differentiation of purified hMDCs was characterized. These cells may be potential biomaterials for human tissue regeneration.

We report a locally aggressive collagenous myofibroblastic neoplasm of the mandible in an 18-year-old male. Clinically, the lesion presented with rapid growth and irregular mandibular bone destruction. Grossly, the tumor was 10 cm in greatest dimension, light-tan, firm, and involving the posterior one-thirds of the body and inferior half of the left mandibular ramus. Histologically, the lesion was composed of a loose spindle cell proliferation interspersed with periodic dense bands of collagen. The spindle cells reacted positively to smooth muscle actin, calponin, and focally to desmin and were negative for S-100, pan-cytokeratin, CD99, CD34 and caldesmon, supporting myofibroblastic derivation. At our 4 year follow-up, the patient remained free of local recurrence and surgery related complications. The clinicopathologic findings and the differential diagnosis of this lesion is presented and discussed.

To investigate the existence of pericryptal fibroblasts sheath (PCFS) in normal gastric mucosa, intestinal metaplasia (IM), indefinite for dysplasia (I-Dys), low grade dysplasia (L-Dys), high grade dysplasia (H-Dys) and gastric cancer (GC), and its association with gastric carcinogenesis. In this study, we examined the existence of PCFS in normal gastric mucosa (N=10), IM (N=26), I-Dys (N=16), L-Dys (N=13), H-Dys (N=21) and GC (N=145) using immunohistochemical staining for two smooth muscle markers, alpha smooth muscle actin(α-SMA) and high molecular weight caldesmon (h-CD). The significance of PCFS was discussed, especially in association with gastric carcinogenesis. The PCFS was recognized in 65.4%(17/26) of IM, 62.5%(10/16) of I-Dys and 23.1% (3/13) of L-Dys respectively. No PCFS was detected in H-Dys and GC. The PCFS was gradually reduced in IM, Dys and GC in sequence (P

Reticular cells and follicular dendritic cells (FDCs) build up a framework that underlies the compartmentalization of spleens and lymph nodes. Subpopulations of reticular cells express the smooth-muscle isoform of actin, indicative of a specialized contractile apparatus. We have investigated the distribution of the actin-binding protein caldesmon in spleen and lymph nodes of mice and rats. Caldesmon modulates contraction and regulates cell motility. Alternative splicing of transcripts from a single gene results in high-molecular-mass isoforms (h-caldesmon) that are predominately expressed by smooth-muscle cells (SMCs), and low-molecular-mass isoforms (l-caldesmon) that are thought to be widely distributed in non-muscle tissues, but the distribution of caldesmon in spleen and lymph nodes has not been reported. We have performed Western blot analysis and immunohistochemistry using four different antibodies against caldesmon, among these a newly developed polyclonal antibody directed against recombinant mouse caldesmon. Western blot analysis showed the preponderance of l-caldesmon in spleen and lymph nodes. Our results from immunohistochemistry demonstrate caldesmon in SMCs, as expected, but also in reticular cells and FDCs, and suggest that the isoform highly expressed by reticular cells is l-caldesmon. In spleen of SCID mice, caldesmon was expressed by reticular cells in the absence of lymphocytes. (J Histochem Cytochem 58:183–193, 2010)

Proprotein convertase 5/6 (PC6), a member of the proprotein convertase (PC) family, is a critical regulator in the uterus for embryo implantation. In particular, PC6 is essential for the differentiation of uterine stromal fibroblasts into decidual cells (decidualization). Knockdown of PC6 in the mouse uterus leads to complete failure of decidualization and implantation. It has been envisaged that PC6 functions by proteolytically activating a number of important growth factors and other precursor proteins. However, to date, the precise mechanism of PC6 action in the uterus, particularly during decidualization, is unknown. In this study, we aimed to investigate the mechanisms of PC6 action in decidualization by identifying its physiological substrates using a proteomic approach. Primary human endometrial stromal cells were decidualized and treated with or without recombinant human PC6 (rhPC6). The proteins cleaved by rhPC6 were identified by 2-dimensional fluorescent differential gel electrophoresis. The candidate proteins were validated as PC6 substrates by a number of approaches, including determining their coexpression with PC6 in vivo in decidual cells in the human uterus. A total of 18 protein spots were significantly altered by rhPC6 treatment, 8 of which were assigned clear identities by mass spectrometry. One of these was confirmed to be caldesmon, a key protein involved in organizing the actin microfilaments and regulating cytoskeletal structure. This study demonstrates a novel approach to identify PC-regulated proteins of physiological relevance, and provides important insight into the mechanism by which PC6 regulates decidualization.

BACKGROUND: Leiomyosarcoma (LMS) can be categorized into uterine, retroperitoneal, nonretroperitoneal soft tissue, cutaneous, visceral, and osseous anatomic subtypes. The differential expression of smooth muscle markers, estrogen receptor (ER), progesterone receptor (PR), and Wilms tumor-1 protein (WT1) by anatomic subtype and gender was explored. METHODS: A total of 78 LMS comprised of 30 uterine and 48 nonuterine tumors were studied. Nonuterine tumors were comprised of 17 soft tissue, 16 retroperitoneal, 7 cutaneous, 5 visceral, and 3 osseous subtypes. Immunohistochemical staining intensity on tissue microarray slides was scored as 0, 1+, or 2+, and cluster analysis was performed on the data. RESULTS: Smooth muscle actin was the most sensitive antibody (95%), followed by muscle-specific actin (91%), calponin (88%), desmin (73%), caldesmon (66%), and myosin (64%). Caldesmon and myosin were usually coexpressed, and were highest in retroperitoneal tumors (94%). There was no discernable correlation noted between histologic differentiation and smooth muscle marker expression. ER was much more common in women, with the highest frequencies noted in female retroperitoneal (86%) and uterine (63%) tumors. Nuclear WT1 was expressed in 11% of all tumors, and was limited to ER-positive uterine and female retroperitoneal tumors. Cluster analysis segregated 4 groups, most notably 1 driven by ER and PR, with the vast majority being uterine and female retroperitoneal tumors. CONCLUSIONS: Smooth muscle markers demonstrated variable sensitivities in LMS, with a tendency for anatomic subtypes to segregate based on expression patterns of these markers. ER defined a subgroup of uterine and female retroperitoneal tumors, and WT1 was limited to such tumors, suggesting a common line of differentiation as well as potential therapeutic targets.

Adenomatoid tumors of the female and male genital tracts are well characterized as mesothelial in origin, but a detailed histological and immunohistochemical analysis comparing both traditional and newer mesothelial markers across gender and site has not been formally conducted. A variety of morphologic features previously described as characteristic of adenomatoid tumors were evaluated in 44 adenomatoid tumors from the male and female genital tracts. Immunohistochemical analysis with pankeratin (AE1/CAM5.2), WT-1, calretinin, CK5/6, D2-40, and caldesmon was also performed. The extent and intensity of staining were scored semiquantitatively on one representative section per case and mean value for each parameter was calculated. All (n=44) the adenomatoid tumors from both the female and male genital tracts demonstrated a distinctive thread-like bridging strand pattern. Lymphoid aggregates were seen in all 12 adenomatoid tumors of male patients, but in only 4 of 32 (13%) tumors in female patients (P

Human adipose stem cells were cultured in smooth muscle inductive media and seeded into synthetic bladder composites to tissue engineer bladder smooth muscle. 85:15 Poly-lactic-glycolic acid bladder dome composites were cast using an electropulled microfiber luminal surface combined with an outer porous sponge. Cell-seeded bladders expressed smooth muscle actin, myosin heavy chain, calponinin, and caldesmon via RT-PCR and immunoflourescence. Nude rats (n = 45) underwent removal of half their bladder and repair using: (i) augmentation with the adipose stem cell-seeded composites, (ii) augmentation with a matched acellular composite, or (iii) suture closure. Animals were followed for 12 weeks post-implantation and bladders were explanted serially. Results showed that bladder capacity and compliance were maintained in the cell-seeded group throughout the 12 weeks, but deteriorated in the acellular scaffold group sequentially with time. Control animals repaired with sutures regained their baseline bladder capacities by week 12, demonstrating a long-term limitation of this model. Histological analysis of explanted materials demonstrated viable adipose stem cells and increasing smooth muscle mass in the cell-seeded scaffolds with time. Tissue bath stimulation demonstrated smooth muscle contraction of the seeded implants but not the acellular implants after 12 weeks in vivo. Our study demonstrates the feasibility and short term physical properties of bladder tissue engineered from adipose stem cells.

Objectives: Ischemia/reperfusion (I/R) has been shown to be the major etiologic factor in animal models in a variety of bladder dysfunctions. Herein we investigate the direct effect of I/R on rabbit bladder contractile regulatory proteins. Methods: Male rabbit bladders were subjected to IR. The contractile responses were recorded and the protein levels of rho-kinase (ROK), caldesmon (CaD) and myosin light chain kinase (MLCK) were analyzed in both bladder muscle and mucosa. Results: For the mucosa layer, ROK was unchanged after ischemia, but increased significantly after 1 week of reperfusion. MLCK increased after ischemia and then significantly increased after 1 and 2 weeks of reperfusion. In the muscle layer, ROK increased at 2 h of reperfusion, decreased significantly following 1 week of reperfusion, then returned to control level by 2 weeks of reperfusion. MLCK expression significantly decreased as early as ischemia alone and did not recover after reperfusion. For CaD expression in the mucosa layer, both CaD isoforms significantly increased at 1 week of reperfusion, then progressively decreased at 2 weeks of reperfusion. In the muscle layer, both CaD isoforms had different expressions. In the muscle layer, smooth muscle caldesmon (h-CaD) decreased at ischemia alone and at 2 h of reperfusion, but significantly increased at 2 weeks of reperfusion; whereas non-muscle caldesmon (l-CaD) significantly decreased at 2 weeks of reperfusion. Conclusion: As bladder muscle and mucosa have markedly different purposes, it is not surprising that they respond differently to I/R. Bladder mucosa and muscle have different kinds of alternations on contractile regulatory proteins.

Distinguishing bladder muscularis propria from muscularis mucosae can be problematic especially in transurethral resection specimens performed for bladder carcinoma. Moreover, bladder carcinoma can be associated with a proliferative/desmoplastic myofibroblastic response that can resemble smooth muscle and potentially lead to overdiagnosis of muscularis propria invasion. The aim of this study was to investigate the potential role of immunohistochemistry in staging bladder carcinoma by evaluating the expression of different markers in myofibroblasts and nonvascular smooth muscle cells in 15 cases of invasive bladder carcinoma. Reactive myofibroblasts were consistently positive for vimentin and smooth muscle actin, consistently negative for caldesmon, desmin, and smoothelin, and had variable expression of actin and CD10. Nonvascular smooth muscle cells of the bladder were consistently positive for smooth muscle actin, actin, desmin, and caldesmon, and consistently negative for CD10. In contrast to smooth muscle cells of the muscularis propria, which displayed strong smoothelin expression in all 15 cases, the smooth muscle cells of the muscularis mucosae displayed moderate smoothelin expression in only 1 (9%) of 11 cases (P=10(-7)). Surprisingly, although strongly highlighting endothelial and endomysial cells, the smooth muscle cells of the muscularis propria weakly expressed vimentin in only 1 (7%) of 15 cases, whereas smooth muscle cells of the muscularis mucosae had moderate or strong expression in 9 (82%) of 11 cases (P=0.00016). The sensitivity and specificity of desmin or caldesmon expression for smooth muscle cells were 100%. The sensitivity and specificity of strong smoothelin expression for muscularis propria were 100%, whereas those of absent vimentin expression were 93 and 82%, respectively. Although morphology remains the gold standard, the findings suggest that immunohistochemistry, using a panel composed of desmin, smoothelin, and vimentin, may be potentially useful for staging of bladder carcinoma. Confirmatory larger-scale studies, especially on transurethral resection specimens, are warranted.

Objectives To detect the effect of ischemia/reperfusion (I/R) injury on rabbit bladder, using physiological study and immunoblotting techniques. Methods Twelve male New Zealand White rabbits were separated into three groups of 4 rabbits each. Group 1 served as control. Group 2 rabbits (ischemia-alone group) underwent in vitro bilateral ischemia surgery for 2 hours. In group 3 (I/R group), bilateral ischemia was similarly induced, and the rabbits were allowed to recover for 2 weeks. The contractile responses to electrical field stimulation, adenosine triphosphate, carbachol, and KCl were recorded. Expression levels of the signaling targets, Rho-kinase (ROK), protein kinase C potentiated inhibitor (CPI-17), caldesmon (CaD), and calponin (CaP) were analyzed by Western blotting. Results Ischemia alone resulted in significant reductions in the contractile responses, whereas I/R resulted in further decreases after all forms of stimulation. In muscle layer, ROK expression increased immediately after ischemia and recovered to the control level after 2 weeks' recovery. However, in mucosa layer, ROK expression showed no significant change after ischemia but significantly increased after reperfusion. After ischemic damage, CPI-17, the functional protein involved in smooth-muscle Ca 2+ sensitization, was significantly increased and then decreased after 2 weeks of reperfusion. The expression of CaP significantly increased after ischemia and decreased after reperfusion. Levels of high-molecular-weight CaD significantly decreased after ischemia and remained very low after reperfusion. Conclusions This study provides further understanding of the role of regulatory proteins in detrusor muscle after ischemia and I/R-induced contractile dysfunction.

The potential roles of the regulatory proteins actin, tropomyosin (Tm), and caldesmon (CaD), i.e., the components of the thin filament, in smooth muscle have been extensively studied in several types of smooth muscles. However, controversy remains on the putative physiological significance of these proteins. In this study, we intended to determine the functional roles of Tm and CaD in the regulation of smooth muscle contraction by using a reconstitution system of the thin filaments. At appropriate conditions, the thin (actin) filaments within skinned smooth muscle strips of taenia caeci in guinea pigs could be selectively removed by an actin-severing protein, gelsolin, without irreversible damage to the contractile apparatus, and then the thin filaments were reconstituted with purified components of thin filaments, i.e., actin, Tm, and CaD. We found that the structural remodeling of actin filaments or thin filaments was functionally linked to the Ca2+-induced force development and reduction in muscle cross-sectional area (CSA). That is, after the reconstitution of the gelsolin-treated skinned smooth muscle strips with pure actin, the Ca2+-dependent force development was partially restored, but the Ca2+-induced reduction in CSA occurred once. In contrast, the reconstitution with actin, followed by Tm and CaD, restored not only the force generation but also both its Ca2+sensitivity and the reversible Ca2+-dependent reduction in CSA. We confirmed that both removal of the thin filaments by gelsolin treatment and reconstitution of the actin (thin) filaments with Tm and CaD caused no significant changes in the level of myosin regulatory light chain phosphorylation. We thus conclude that Tm and CaD are necessary for the full regulation of smooth muscle contraction in addition to the other regulatory systems, including the myosin-linked one.

The purpose of this study is to compare vaginal caldesmon expression in women with and without anterior vaginal wall prolapse.Vaginal tissues were sampled in women with (n = 11) or without (n = 11) vaginal wall prolapse. Caldesmon messenger RNA (mRNA) expression was assessed by quantitative real-time polymerase chain reaction. Immunohistochemistry and digital image analysis were used to determine caldesmon protein expression in the histologic sections.There were no significant differences in demographic data between the two groups. Caldesmon mRNA expression was significantly decreased in the vaginal tissue from women with anterior vaginal wall prolapse compared to women without prolapse [(caldesmon mean +/- SD mRNA expression in relative units) 0.03 +/- 0.03 vs 0.17 +/- 0.17, P = 0.02]. The fractional area of nonvascular caldesmon staining in the vagina of women with anterior vaginal wall prolapse was significantly decreased compared to women without prolapse [mean +/- SD (0.09 +/- 0.04 vs 0.16 +/- 0.09, P = 0.03)].Vaginal caldesmon expression is significantly decreased in women with anterior vaginal wall prolapse compared to normal subjects.

The zebrafish homologue of caldesmon is similar to the mammalian low molecular weight caldesmon (l-CaD). In this study, we explored the effects of caldesmon knockdown on vertebrate heart development in vivo. In a zebrafish model caldesmon was knocked down resulting in defective cardiac morphogenesis, muscularization and function. The data provide the first functional assessment of the role of caldesmon in cardiac development in vivo, and indicate that caldesmon is essential for proper cardiac organogenesis and function. Because caldesmon expression remarkably influences cardiac muscularization, the findings are relevant for designing future therapeutic strategies in the regeneration of cardiac damage. (C) 2008 Elsevier Inc. All rights reserved

Previous studies have shown that microgroove-initiated contact guidance can induce bone formation in osteoprogenitor cells (OPGs) and produce changes in the cell proteome. For proteomic analysis, differential in-gel electrophoresis (DIGE) can be used as a powerful diagnostic method to provide comparable data between the proteomic profiles of cells cultured in different conditions. This study focuses on the response of OPGs to a novel nanoscale pit topography with osteoinductive properties compared with planar controls. Disordered near-square nanopits with 120 nm diameter and 100 nm depth with an average 300 nm centre-to-centre spacing (300 nm spaced pits in square pattern, but with ±50 nm disorder) were fabricated on 1×1 cm2polycaprolactone sheets. Human OPGs were seeded onto the test materials. DIGE analysis revealed changes in the expression of a number of distinct proteins, including upregulation of actin isoforms, beta-galectin1, vimentin and procollagen-proline, 2-oxoglutarate 4-dioxygenase and prolyl 4-hydroxylase. Downregulation of enolase, caldesmon, zyxin, GRASP55, Hsp70 (BiP/GRP78), RNH1, cathepsin D and Hsp27 was also observed. The differences in cell morphology and mineralization are also reported using histochemical techniques.

The effect of the suppression of expression of the actin-binding protein caldesmon on the motility of nonmuscle cells has been studied. A more than a fivefold decrease in the content of this protein in cells by RNA interference led to the disturbance of the formation of actin stress fibers and acceleration of cell migration to the zone of injury of the monolayer. A stimulation of stationary cells by serum induced more than 1,5-fold accumulation of stress fibers only in control cells, but not in caldesmon-deficient cells. Similarly, the accumulation of actin filaments was observed in actively migrating cells of only wild type, but not in the cells with low caldesmon content. These changes occurred mainly at the leading edge of the migrating cell where the distinct structure of actin filaments was not seen in the absence of caldesmon. It was assumed that caldesmon inhibits cell migration due to the stabilization of actin in filaments and a decrease in the dynamics of monomeric actin at the leading edge of the migrating cell.

We have previously described differential effects of physiologic (5%) and pathologic (18%) cyclic stretch (CS) on agonist-induced pulmonary endothelial permeability. This study examined acute and chronic effects of CS on agonist-induced intracellular signaling and cell morphology in the human lung macro- and microvascular endothelial cell (EC) monolayers. Endothelial permeability was assessed by analysis of morphological changes, parameters of cell contraction and measurements of transendothelial electrical resistance. Exposure of both microvascular and macrovascular EC to 18% CS for 2 – 96 hrs increased thrombin-induced permeability and monolayer disruption. Interestingly, the ability to promote thrombin responses was present in EC cultures exposed to 48–96 hrs of CS even after replating onto non-elastic substrates. In turn, physiologic CS preconditioning (72 hrs) attenuated thrombin-induced paracellular gap formation and MLC phosphorylation in replated EC cultures. Long term preconditioning at 18% CS (72 hrs) increased the content of signaling and contractile proteins including Rho GTPase, MLC, MLC kinase, ZIP kinase, PAR1, caldesmon and HSP27 in the pulmonary microvascular and macrovascular cells. We conclude that short term CS regulates EC permeability via modulation of agonist-induced signaling, whereas long term CS controls endothelial barrier at both post-translational level and via magnitude-dependent regulation of pulmonary EC phenotype, signaling and contractile protein expression.

Smooth muscle contractile activity is a major regulator of function of the vascular system, respiratory system, gastrointestinal system and the genitourinary systems. Malfunction of contractility in these systems leads to a host of clinical disorders, and yet, we still have major gaps in our understanding of the molecular mechanisms by which contractility of the differentiated smooth muscle cell is regulated. This review will summarize recent advances in the molecular understanding of the regulation of smooth muscle myosin activity via phosphorylation/dephosphorylation of myosin, the regulation of the accessibility of actin to myosin via the actin-binding proteins calponin and caldesmon, and the remodelling of the actin cytoskeleton. Understanding of the molecular ‘players’ should identify target molecules that could point the way to novel drug discovery programs for the treatment of smooth muscle disorders such as cardiovascular disease, asthma, functional bowel disease and pre-term labour.

Purpose The internal anal sphincter (IAS) plays an important role in the pathophysiology of constipation and incontinence. We hypothesized that functional bowel obstruction in premature infants is because of a poorly developed IAS. We investigated the neuromuscular development of IAS in fetal, newborn, and adolescent pigs. Methods Paraffin sections of IAS from 5 different age groups, E60, E90, 1 day, 4, and 12 weeks old, were stained with protein gene product 9.5 (PGP9.5), α -smooth muscle actin ( α -SMA), caldesmon (CALD), calponin (CALP), and desmin (DES) antibodies. Quantification of results was performed by grading the density of immunostaining. Results The PGP9.5-positive ganglion cells were observed in the myenteric and submucosal region of the entire length of the IAS at E60. An increase in ganglion cell size and density was observed with increasing age. There were striking differences in the density of PGP9.5, α -SMA, DES, CALD, and CALP immunoreactive fibers between prenatal and postnatal period with gradient increase in the number of fibers from after birth to 12 weeks of age. Conclusion This study shows for the first time that there are age-related differences in the distribution of neurons and smooth muscle cell components in the IAS. The decreased expression of contractile and cytoskeleton proteins in smooth muscle cells together with decreased expression of neurons in the IAS in the perinatal period may lead to motility dysfunction causing functional intestinal obstruction seen in premature infants.

Lysenin, a toxin present in the coelomic fluid of the earthworm Eisenia foetida, is known to cause a long-lasting contraction of rat aorta smooth muscle strips. We addressed the mechanisms underlying its action on smooth muscle cells and present the first report demonstrating a completely new property of lysenin unrelated to its basic sphingomyelin-binding ability. Here we report lysenin enhancement effect on smooth muscle actomyosin ATPase activity and the ability of networking the actin filaments. The maximum enhancement of the ATPase activity of actomyosin at 120 mM KCl was observed at a molar ratio of lysenin to actin of about 1:10(5), while at 70 mM KCl at the ratio of about 1:10(6). The effect of lysenin became most pronounced only when both smooth muscle regulatory proteins, tropomyosin and caldesmon, were present. Co-sedimentation experiments indicated that lysenin did not displace neither tropomyosin nor caldesmon from the thin filament. Thus, the lysenin-dependent abolishment of the inhibitory effect of caldesmon on the ATPase activity was related rather to the modification of the filament structure. The ability of the toxin to exert its stimulatory effect at extremely low concentrations (as low as one molecule of lysenin per 10(6) actin molecules) may result from the long-range cooperative transitions in the entire thin filament with an involvement of smooth muscle tropomyosin, while the role of caldesmon may be limited exclusively to the inhibition of ATPase activity.

Diabetic nephropathy (DN) develops in about 40% of insulin-dependent type 1 diabetes mellitus (T1DM) patients, and is associated not only with diabetes duration and metabolic control, but also with a genetic predisposition. Constitutive alterations of cytoskeletal proteins may play a role in the development of DN. We investigated the expression of these proteins in cultured skin fibroblasts, obtained from long-term T1DM patients with and without DN but comparable metabolic control, and from matched healthy subjects, by means of 2-DE electrophoresis and MS-MALDI analyses. In T1DM with DN, compared to the other two groups, quantitative analyses revealed an altered expression of 17 spots (p

Smooth muscle thin filaments are made up of actin, tropomyosin, caldesmon, and a Ca2+-binding protein and their interaction with myosin is Ca2+-regulated. We suggested that Ca2+ regulation by caldesmon and Ca2+-calmodulin is achieved by controlling the state of thin filament through a cooperative-allosteric mechanism homologous to troponin-tropomyosin in striated muscles. In the present work, we have tested this hypothesis. We monitored directly the thin filament transition between the ON and OFF state using the excimer fluorescence of pyrene iodoacetamide (PIA)-labeled smooth muscle αα-tropomyosin homodimers. In steady state fluorescence measurements, myosin subfragment 1 (S1) cooperatively switches the thin filaments to the ON state, and this is exhibited as an increase in the excimer fluorescence. In contrast, caldesmon decreases the excimer fluorescence, indicating a switch of the thin filament to the OFF state. Addition of Ca2+-calmodulin increases the excimer fluorescence, indicating a switch of the thin filament to the ON state. The excimer fluorescence was also used to monitor the kinetics of the ON-OFF transition in a stopped-flow apparatus. When ATP induces S1 dissociation from actin-PIA-tropomyosin, the transition to the OFF state is delayed until all S1 molecules are dissociated actin. In contrast, caldesmon switches the thin filament to the OFF state in a cooperative way, and no lag is displayed in the time course of the caldesmon-induced fluorescence decrease. We have also studied caldesmon and Ca2+-calmodulin-caldesmon binding to actin-tropomyosin in the ON and OFF states. The results are used to discuss both caldesmon inhibition and Ca2+-calmodulin-caldesmon activation of actin-tropomyosin.

Backgrounds: Immature teratoma (IT) of vulva (V) has been reported only once. Case report: A 15-year-old girl presented a vulvar tumor, and tumorectomy was performed. The tumor measured 4 cm × 4 cm × 5 cm and multiply cystic. The cysts contained mucinous fluid. The tumor was well demarcated from surrounding tissues and tumorectomy was easy. Microscopically, the tumor was multicystic neoplasm composed of mucin-bearing columnar mature epithelial cells, cellular immature mesenchymal zone beneath the columnar epithelial cells, immature cartilage, and mature connective tissue containing vasculatures. Immunohistochemically, the immature mesenchymal elements of the tumor were positive for p53, Ki67 (labeling = 30%) and vimentin, but negative for smooth muscle actin, myoglobin, HHF-35 caldesmon, and S100 element. No immature neural elements were seen. Since three germ elements were seen and some showed immature features, a diagnosis of IT was made. Conclusions: A very rare case of IT of V is reported. This is the second case of IT in this location.

Background: The primary risk factor for primary-open angle glaucoma (POAG) is increased intraocular pressure (IOP). In POAG patients, the outflow resistance through the trabecular meshwork (TM) is abnormally elevated. One of the important glaucoma-associated pathological changes in the TM is formation of excessive cross-linked actin networks (CLANs). CLANs are web-like polygonal structures found in confluent glaucoma TM cells and tissues. Glaucoma-associated factors transforming forming growth factor beta 2 (TGFβ2) and glucocorticoids induce CLAN formation in non-glaucoma TM cells (NTM). CLANs may increase cell stiffness and alter homeostasis, and thereby contribute to elevated IOP. Methods: We used a proteomic approach to identify CLAN-associated proteins. We treated confluent primary NTM cells with 0.1% ethanol (EtOH; vehicle), 100 nM dexamethasone (DEX), or 5 ng/ml TGFβ2 plus 0.1% EtOH for 7 days to induce CLANs. The Triton insoluble fraction containing cytoskeleton was extracted for two-dimensional differential in-gel electrophoresis (2D-DIGE). Mass spectrometry (MS) was used to identify the differential expressed proteins in the 2D-gels. Co-localization of identified proteins with CLANs was confirmed using immunocytofluorescence (ICF) microscopy. Results: 2D-DIGE revealed 103 differentially expressed proteins in both treatment groups. MSidentified 23 of the most enriched proteins. ICF showed that caldesmon, calponin, myosin light chain, and tropomyosin were colocalized with CLANs. Conclusions: We identified a subset of proteins differentially expressed in NTM cells with DEX or TGFβ2- induced CLAN formation. The potential involvement of these proteins in CLAN formation and/or maintenance requires further investigation. These proteins may provide new insights into the pathogenesis of glaucoma.

Tamoxifen therapy resistance constitutes a major cause of death in patients with recurrent estrogen receptor (ER) positive breast cancer. Through high resolution mass spectrometry (MS), we previously generated a 4-protein predictive signature for tamoxifen therapy outcome in recurrent breast cancer. ANXA1 and CALD1, which were not included in the classifier, were however the most differentially expressed proteins. We first evaluated the clinical relevance of these markers in our MS cohort, followed by immunohistochemical (IHC) staining on an independent set of tumors incorporated in a tissue microarray (TMA) and regression analysis in relation to time to progression (TTP), clinical benefit and objective response. In order to assess which mechanisms ANXA1 and CALD1 might been involved in, we performed Ingenuity pathway analysis (IPA) on ANXA1 and CALD1 correlated proteins in our MS cohort. ANXA1 (Hazard ratio [HR] = 1.83; 95% confidence interval [CI]: 1.22-2.75; P = 0.003) and CALD1 (HR = 1.57; 95% CI: 1.04-2.36; P = 0.039) based patient stratification showed significant association to TTP, while IHC staining on TMA showed that both ANXA1 (HR = 1.82; 95% CI: 1.12-3.00; P = 0.016) and CALD1 (HR = 2.29; 95% CI: 1.40-3.75; P = 0.001) expression was associated with shorter TTP independently of traditional predictive factors. Pearson correlation analysis showed that the majority of proteins correlated to ANXA1 also correlated with CALD1. IPA indicated that ANXA1 and CALD1 were associated with ER-downregulation and NF kappa B signaling. We hereby report that ANXA1 and CALD1 proteins are independent markers for tamoxifen therapy outcome and are associated to fast tumor progression.

Çalışmada 21 yazar bulunmaktadır. Bu yazarlardan sadece Bursa Uludağ Üniversitesi mensuplarının girişleri yapılmıştır. Vascular malformations are non-neoplastic expansions of blood vessels that arise due to errors during angiogenesis. They are a heterogeneous group of sporadic or inherited vascular disorders characterized by localized lesions of arteriovenous, capillary, or lymphatic origin. Vascular malformations that occur inside bone tissue are rare. Herein, we report loss-of-function mutations in ELMO2 (which translates extracellular signals into cellular movements) that are causative for autosomal-recessive intraosseous vascular malformation (VMOS) in five different families. Individuals with VMOS suffer from life-threatening progressive expansion of the jaw, craniofacial, and other intramembranous bones caused by malformed blood vessels that lack a mature vascular smooth muscle layer. Analysis of primary fibroblasts from an affected individual showed that absence of ELMO2 correlated with a significant downregulation of binding partner DOCK1, resulting in deficient RAC1-dependent cell migration. Unexpectedly, elmo2-knockout zebrafish appeared phenotypically normal, suggesting that there might be human-specific ELMO2 requirements in bone vasculature homeostasis or genetic compensation by related genes. Comparative phylogenetic analysis indicated that elmo2 originated upon the appearance of intramembranous bones and the jaw in ancestral vertebrates, implying that elmo2 might have been involved in the evolution of these novel traits. The present findings highlight the necessity of ELMO2 for maintaining vascular integrity, specifically in intramembranous bones. CRANIRARE consortium - R07197KS Strategic Positioning Fund for Genetic Orphan Diseases Agency for Science Technology & Research (A*STAR) Hacettepe Üniversitesi- 00-02-101-009 / 03-D07-101-001

Background and aims Proprotein convertase subtilisin kexin type 9 (PCSK9) induces degradation of the low-density lipoprotein-receptor (LDLR). Smooth muscle cells (SMCs) in human atherosclerotic plaques and cultured SMCs express PCSK9. The present study aimed at defining the role of PCSK9 on vascular response to injury. Methods Carotid neointimal lesions were induced by positioning a non-occlusive collar in PCSK9 knock-out ( PCSK9 −/− ) and wild type littermate ( PCSK9 +/+ ) mice. Results In PCSK9 −/− mice, we observed a significantly less intimal thickening ( p p PCSK9 +/+ mice. When compared with PCSK9 −/− , lesions of PCSK9 +/+ mice had a higher content of SMCs ( p p PCSK9 −/− , when compared to PCSK9 +/+ cells, showed higher levels of α-smooth muscle actin ( α-SMA ; 2.24 ± 0.36 fold; p MHC-II ; 8.65 ± 1.55 fold; p caldesmon mRNA(−54 ± 14%; p PCSK9 −/− cells also showed a slower proliferation rate, and an impaired migratory capacity and G1/S progression of the cell cycle. The reconstitution of PCSK9 expression, by retroviral infection of PCSK9 −/− SMCs, led to a downregulation of α-SMA (−56 ± 2%; p MHC-II (−45% ± 25.5 fold: p = 0.06) and calponin (−25% ± 0.8 fold: p caldesmon mRNA (1.46 ± 0.3 fold; p PCSK9 −/− was significantly lower compared to PCSK9 reconstituted cells. Conclusions Taken together, the present results suggest that PCSK9, by sustaining SMC synthetic phenotype, proliferation, and migration, may play a pro-atherogenic role in the arterial wall.

Venöz invazyon, tüm tümörlerde bağımsız bir prognostik faktör olup özellikle evre II hastalarda adjuvan kemoterapi kararını etkileyebilmektedir.Bu çalışmada, CD31, SMM, transgelin ve smoothelin antikorlarının, gastrik, pankreatik ve kolorektal adenokarsinomlarda venöz invazyonun gösterilmesinde birbirlerine üstünlüklerinin karşılaştırılması amaçlanmıştır.Gazi Üniversitesi Tıbbi Patoloji Anabilim Dalı'nda 2007- 2015 yılları arasında tanı alan 11 Whipple, 5 total gastrektomi, 3 segmental kolektomi ve 1 low anterior rektum rezeksiyon materyaline ait 79 blokta bulunan toplam 254 odak ele alınmıştır. Yirmi hastaya ait yaş, cinsiyet, tümör yerleşim yeri, tümör çapı, histolojik tipi ve derecesi, T, N, M değerleri incelenmiştir. Olası Vİ odaklarının anatomik lokalizasyonu, histolojik özellikleri gözden geçirilmiş ve bu odaklar 3 tipe ayrılmıştır: A tipi `orphan arteriol` paterni, F tipi, eşlik eden arter yapısı olmadan, organ çevresindeki yağ dokuda serbest tümör depozitleri, X tipi, uygulanan immünhistokimyasal yöntem sonrası ortaya çıkan odaklar.Çalışmamızda 254 odağın %59,8'i A tipi, %29,1'i X tipi ve %11'i F tipidir. A tipi odakların %87,5'i, X tipi odakların %13,5'i ekstramuraldir. F tipi odaklar tanımı gereği tümü ile ekstramuraldir. İmmünhistokimyasal belirteçler öncelikle negatif (0) ve pozitif (1) olarak değerlendirilmiş, pozitif olanlar, damar yapısının çembersel çevresini pozitif boyama yeteneğinin niceliğine göre +1, +2 ve +3 olarak skorlanmıştır.Çalışmamızın sonucunda, olası Vİ odaklarının hiçbirinde smoothelin ve CD31 ile ekspresyon saptanmamıştır. Tüm odaklar toplu halde ve A ve F odakları ayrı olarak incelendiğinde transgelinin SMM'ye göre daha üstün bir belirteç olduğu görülmüştür (sırasıyla p