Endothelial Monocyte-activating Polypeptide Research Articles

The effect of eight weeks of high-intensity interval training on the expression of mir-126, EMAP-II, and levels of TGF-β in the heart tissue of young rats with type 2 diabetes

Background and aims: Diabetes affects some cardiac angiogenic and antiangiogenic markers. Thus, the aim of this study was to investigate the effect of eight weeks of high-intensity interval training (HIIT) on mir-126, endothelial monocyte-activating polypeptide II (EMAP-II), and transforming growth factor beta (TGF-β) in young rats with type 2 diabetes (T2D). Methods: The statistical population of this experimental study included young rats (8 weeks old) with T2D. In this research, 24 young rats (8 weeks old) were divided into healthy+HIIT, diabetic+HIIT, and healthy control groups. The enzyme-linked immunosorbent assay method was used to measure TGF-β and the real-time polymerase chain reaction for mir-126 and EMAP-II. A one-way analysis of variance and the least significant difference post hoc test were used to compare the two groups. Results: The results showed diabetes+HIIT and healthy+HIIT had more mir-126 compared to healthy control (P=0.001). Healthy+HIIT, in comparison to diabetes+HIIT, had higher mir-126 expression (P=0.001). In addition, diabetes+HIIT, compared to the healthy control, had a higher EMAP-II expression (P=0.006). On the other hand, healthy+HIIT had lower EMAP-II in comparison to healthy control (P=0.001). Healthy+HIIT had a lower EMAP-II compared to diabetes+HIIT (P=0.001). Diabetes+HIIT, in comparison to the healthy control, had higher TGF-β levels (P=0.005). However, healthy+HIIT, compared to healthy control, had a lower expression of TGF-β (P=0.001). Finally, the healthy+HIIT groups, compared to the diabetes+HIIT group, had a lower TGF-β expression (P=0.001). Conclusion: Interval exercise could increase angiogenesis in diabetic samples through the increase of mir-126 and the reduction of TGF-β1 and EMAP-II.

Altered Smooth Muscle Cell Histone Acetylome by the SPHK2/S1P Axis Promotes Pulmonary Hypertension.

Epigenetic regulation of vascular remodeling in pulmonary hypertension (PH) is poorly understood. Transcription regulating, histone acetylation code alters chromatin accessibility to promote transcriptional activation. Our goal was to identify upstream mechanisms that disrupt epigenetic equilibrium in PH. Human pulmonary artery smooth muscle cells (PASMCs), human idiopathic pulmonary arterial hypertension (iPAH):human PASMCs, iPAH lung tissue, failed donor lung tissue, human pulmonary microvascular endothelial cells, iPAH:PASMC and non-iPAH:PASMC RNA-seq databases, NanoString nCounter, and cleavage under targets and release using nuclease were utilized to investigate histone acetylation, hyperacetylation targets, protein and gene expression, sphingolipid activation, cell proliferation, and gene target identification. SPHK2 (sphingosine kinase 2) knockout was compared with control C57BL/6NJ mice after 3 weeks of hypoxia and assessed for indices of PH. We identified that Human PASMCs are vulnerable to the transcription-promoting epigenetic mediator histone acetylation resulting in alterations in transcription machinery and confirmed its pathological existence in PH:PASMC cells. We report that SPHK2 is elevated as much as 20-fold in iPAH lung tissue and is elevated in iPAH:PASMC cells. During PH pathogenesis, nuclear SPHK2 activates nuclear bioactive lipid S1P (sphingosine 1-phosphate) catalyzing enzyme and mediates transcription regulating histone H3K9 acetylation (acetyl histone H3 lysine 9 [Ac-H3K9]) through EMAP (endothelial monocyte activating polypeptide) II. In iPAH lungs, we identified a 4-fold elevation of the reversible epigenetic transcription modulator Ac-H3K9:H3 ratio. Loss of SPHK2 inhibited hypoxic-induced PH and Ac-H3K9 in mice. We discovered that pulmonary vascular endothelial cells are a priming factor of the EMAP II/SPHK2/S1P axis that alters the acetylome with a specificity for PASMC, through hyperacetylation of histone H3K9. Using cleavage under targets and release using nuclease, we further show that EMAP II-mediated SPHK2 has the potential to modify the local transcription machinery of pluripotency factor KLF4 (Krüppel-like factor 4) by hyperacetylating KLF4 Cis-regulatory elements while deletion and targeted inhibition of SPHK2 rescues transcription altering Ac-H3K9. SPHK2 expression and its activation of the reversible histone H3K9 acetylation in human pulmonary artery smooth muscle cell represent new therapeutic targets that could mitigate PH vascular remodeling.

Endothelin-1 gene polymorphism (G8002A) and endothelial monocyte-activating polypeptide II: Role in vascular dysfunction in pediatric patients with β-thalassemia major.

Endothelin-1 (ET-1), a potent endogenous vasoconstrictor, stimulates production of reactive oxygen species. Endothelial monocyte-activating polypeptide-II (EMAP-II) is a multifunctional polypeptide. To assess ET-1 gene polymorphism (G8002A) in pediatric patients with β-thalassemia major (β-TM) as a potential genetic marker for vascular dysfunction and its possible relation to EMAP II, oxidative stress and vascular complications. β-TM patients (n=95) without symptomatic cardiac or renal disease were compared with 95 healthy controls. Markers of hemolysis, serum ferritin, urinary albumin-to-creatinine ratio, serum EMAP II, malondialdehyde (MDA) and antioxidant enzymes; superoxide dismutase (SOD), glutathione peroxidase (GPx), reduced glutathione (GSH), glutathione reductase and catalase were measured. ET-1 gene polymorphism (G8002A) was determined using polymerase chain reaction‑restriction fragment length polymorphism. β-TM patients had significantly higher EMAP II than healthy controls. EMAP II was significantly higher among patients with cardiac disease, pulmonary hypertension (PH) risk, nephropathy, poor compliance to therapy and ferritin≥2500μg/L. There were significant correlations between EMAP II and transfusion index, LDH, ferritin and oxidative stress markers. The AA genotype of ET-1 gene polymorphism (G8002A) was significantly higher among β-TM patients than controls. The number of patients with cardiac disease, PH risk or nephropathy was significantly higher among AA genotype compared with GG and GA genotypes. Lactate dehydrogenase (LDH), serum ferritin, EMAP II, MDA, SOD and GPx were significantly higher in AA genotype. ET-1 gene polymorphism (G8002A) could be a possible genetic marker for prediction of increased susceptibility to cardiopulmonary and renal complications among pediatric patients with β-TM.

PROGNOSTICATION OF COMPLICATED COURSE OF ACUTE MYOCARDIAL INFARCTION WITH CONCOMITANT TYPE 2 DIABETES MELLITUS BASED ON USE OF ENDOTHELIAL MONOCYTE ACTIVATING POLYPEPTIDE-ІІ

The aim: To determine the prognostic value of endothelial monocytic peptide ІІ in the course of AMI (acute myocardial infarction) in combination with type 2 diabetes mellitus (DM) Materials and methods: The study involved 120 patients divided in 2 groups: Group 1 – patients with acute myocardial infarction (AMI) with concomitant type 2 DM (n=69); Group 2 – patients with AMI without concomitant type 2 DM (n=51). Control group was composed with 20 almost healthy persons. Results: It was determined that in patients with AMI in combination with type 2 diabetes, the level of endothelial monocyte-activating polypeptide II (EMAP-II) was 1.65 times higher than in patients without concomitant type 2 DM (p <0.05). The patients with AMI and type 2 DM who were included in the group of the 3rd tertile according to level of EMAR-II had the complicated course compared to patients in the groups of the 1st and 2nd tertiles. Q-positive AMI was found in 100% of patients who belonged to the group of the 3rd tertile; recurrence of AMI occurred only in patients whose EMAR-II index was included in the 3rd tertile. Conclusions: According to the results of endothelial function analysis with usу ща the marker of EMAP-II endothelial dysfunction in patients with AMI and concomitant type 2 DM, the increased level of this parameter was characteristic of pronounced violation of dilatation properties of the vascular wall and of the other indicators of complicated comorbid conditions.

Маркери ендотеліальної дисфункції як предиктори ускладненого перебігу гострого інфаркту міокарда у поєднанні з цукровим діабетом 2 типу

Today, the endothelium is considered to be a multifunctional organ and a non-specific marker in the pathogenesis of diseases of the cardiovascular system. The course of diseases of the cardiovascular system depends on the presence of comorbid pathology, where type 2 diabetes is one of the possible representations of such pathology. The purpose of the study was to determine the diagnostic role of marker of endothelial dysfunction in patients with acute myocardial infarction with concomitant type 2 diabetes mellitus with regard to the clinical course of comorbid conditions. Material and methods. The study design consisted of 120 patients. They were divided into 2 groups: Group 1 consisted of patients with acute myocardial infarction and concomitant type 2 diabetes (n=69), Group 2 consisted of patients with acute myocardial infarction without concomitant type 2 diabetes (n=51). Patients of both groups matched on age and sex (60 men (50%) and 60 women (50%); their average age was 66.35±0.91 years, р<0,05). The control group consisted of 20 almost healthy people, among them 12 women (60%) and 8 men (40%) (average age was equal to 45.17±2.88 years). Examination of patients was conducted on the basis of Municipal Non-profit Enterprise "City Clinical Hospital No. 27" of Kharkiv City Council in the department of cardiology for patients with acute myocardial infarction. Diagnoses were determined according to the current criteria. All patients enrolled in the study signed a voluntary informed consent to participate in it. All patients underwent general clinical and instrumental examinations. The level of endothelial monocyte-activating polypeptide II in participants of the study was determined on the first day of acute myocardial infarction by using a commercial test system "Human Endothelial monocyte activating polepeptide II ELISA KIT". Results and discussion. According to the results of the study, in patients with acute myocardial infarction in combination with type 2 diabetes the levels of endothelial monocyte-activating polypeptide II was higher than in patients without concomitant type 2 diabetes by 1.65 times (p <0.05). The course of the study revealed the level of endothelial monocyte-activating polypeptide II, which was marked by high risk of cardiogenic shock and acute left ventricular failure. Having analyzed the parameter of endothelial dysfunction in terms of mortality, the researchers made the following conclusion: the endothelial monocyte-activating polypeptide II values greater than 3.44 ng/ml were the predictor of cardiovascular death probability during 6 months. Conclusion. The results of analysis of endothelial function made with use of the marker of endothelial monocyte-activating polypeptide II in patients with acute myocardial infarction with concomitant type 2 diabetes showed that such indicator can be deemed as predictor in complicated comorbid conditions

Endothelial Monocyte-Activating Polypeptide II Mediates Macrophage Migration in the Development of Hyperoxia-Induced Lung Disease of Prematurity.

Myeloid cells are key factors in the progression of bronchopulmonary dysplasia (BPD) pathogenesis. Endothelial monocyte-activating polypeptide II (EMAP II) mediates myeloid cell trafficking. The origin and physiological mechanism by which EMAP II affects pathogenesis in BPD is unknown. The objective was to determine the functional consequences of elevated EMAP II levels in the pathogenesis of murine BPD and to investigate EMAP II neutralization as a therapeutic strategy. Three neonatal mouse models were used: (1) BPD (hyperoxia), (2) EMAP II delivery, and (3) BPD with neutralizing EMAP II antibody treatments. Chemokinic function of EMAP II and its neutralization were assessed by migration in vitro and in vivo. We determined the location of EMAP II by immunohistochemistry, pulmonary proinflammatory and chemotactic gene expression by quantitative polymerase chain reaction and immunoblotting, lung outcome by pulmonary function testing and histological analysis, and right ventricular hypertrophy by Fulton's Index. In BPD, EMAP II initially is a bronchial club-cell-specific protein-derived factor that later is expressed in galectin-3+ macrophages as BPD progresses. Continuous elevated expression corroborates with baboon and human BPD. Prolonged elevation of EMAP II levels recruits galectin-3+ macrophages, which is followed by an inflammatory state that resembles a severe BPD phenotype characterized by decreased pulmonary compliance, arrested alveolar development, and signs of pulmonary hypertension. In vivo pharmacological EMAP II inhibition suppressed proinflammatory genes Tnfa, Il6, and Il1b and chemotactic genes Ccl2 and Ccl9 and reversed the severe BPD phenotype. EMAP II is sufficient to induce macrophage recruitment, worsens BPD progression, and represents a targetable mechanism of BPD development.

Distribution and characterization of tumor-associated macrophages/microglia in rat C6 glioma

Immunity responses and immunotherapy are novel areas of research for the pathological development and treatment of glioma, the most common brain cancer. Characterization of the subpopulations of infiltrated immune cells may aid in our understanding of the tumor immune response and contribute to the identification of cellular targets for selective immunotherapy. Using a rat C6 glioma model, the present study observed a significant heterogeneity of active macrophages and microglia, including cluster of differentiation 8 (CD8)+, endothelial monocyte-activating polypeptide II (EMAPII)+ and ED1+ cells, mostly in the areas of compact tumor growth and inside or around the pannecrosis. Moreover, the CD8+ cells were similar to reactive ED1+ and EMAPII+ microglia/macrophages in morphology and distribution, but different from the W3/13+ T cells. These observations suggest that different subtypes of macrophages and microglia are involved in glioma development and thus, may be potential targets for immunotherapeutic antitumor strategies.

Endothelial monocyte activating polypeptide II in children and adolescents with type 1 diabetes mellitus: Relation to micro-vascular complications

Objectives: Endothelial monocyte-activating polypeptide II (EMAP II) is a multifunctional polypeptide with proinflammatory and antiangiogenic activity. Hyperglycemia and dyslipidemia appears to be significant factors contributing to increased EMAP-II levels. We determined serum EMAP II in children and adolescents with type 1 diabetes as a potential marker for micro-vascular complications and assessed its relation to inflammation and glycemic control. Methods: Eighty children and adolescents with type 1 diabetes were divided into 2 groups according to the presence of micro-vascular complications and compared with 40 healthy controls. High-sensitivity C-reactive protein (hs-CRP), hemoglobin A1c (HbA1c) and EMAP II levels were assessed. Results: Serum EMAP II levels were significantly increased in patients with micro-vascular complications (1539±321.5pg/mL) and those without complications (843.6±212.6pg/mL) compared with healthy controls (153.3±28.3pg/mL; p<0.001). EMAP II was increased in patients with microalbuminuria than normoalbuminuric group (p<0.001). Significant positive correlations were found between EMAP II levels and body mass index, fasting blood glucose, HbA1c, serum creatinine, triglycerides, total cholesterol, urinary albumin creatinine ratio (UACR) and hs-CRP (p<0.05). A cutoff value of EMAP II at 1075pg/mL could differentiate diabetic patients with and without micro-vascular complications with a sensitivity of 93% and specificity of 82%. Conclusions: We suggest that EMAP II is elevated in type 1 diabetic patients, particularly those with micro-vascular complications. EMAP II levels are related to inflammation, glycemic control, albuminuria level of patients and the risk of micro-vascular complications.

HIV envelope protein gp120-induced apoptosis in lung microvascular endothelial cells by concerted upregulation of EMAP II and its receptor, CXCR3

Chronic lung diseases, such as pulmonary emphysema, are increasingly recognized complications of infection with the human immunodeficiency virus (HIV). Emphysema in HIV may occur independent of cigarette smoking, via mechanisms that are poorly understood but may involve lung endothelial cell apoptosis induced by the HIV envelope protein gp120. Recently, we have demonstrated that lung endothelial apoptosis is an important contributor to the development of experimental emphysema, via upregulation of the proinflammatory cytokine endothelial monocyte-activating polypeptide II (EMAP II) in the lung. Here we investigated the role of EMAP II and its receptor, CXCR3, in gp120-induced lung endothelial cell apoptosis. We could demonstrate that gp120 induces a rapid and robust increase in cell surface expression of EMAP II and its receptor CXCR3. This surface expression occurred via a mechanism involving gp120 signaling through its CXCR4 receptor and p38 MAPK activation. Both EMAP II and CXCR3 were essentially required for gp120-induced apoptosis and exposures to low gp120 concentrations enhanced the susceptibility of endothelial cells to undergo apoptosis when exposed to soluble cigarette smoke extract. These data indicate a novel mechanism by which HIV infection causes endothelial cell loss involved in lung emphysema formation, independent but potentially synergistic with smoking, and suggest therapeutic targets for emphysema prevention and/or treatment.

Monoclonal Antibodies Against Tyrosyl-tRNA Synthetase and Its Isolated Cytokine-Like Domain

Tyrosyl-tRNA synthetase (TyrRS) is one of the key enzymes of protein biosynthesis. In addition to its basic role, this enzyme reveals some important non-canonical functions. Under apoptotic conditions, the full-length enzyme splits into two fragments having distinct cytokine activities, thereby linking protein synthesis to cytokine signaling pathways. The NH2-terminal catalytic fragment, known as miniTyrRS, binds strongly to the CXC-chemokine receptor CXCR1 and, like interleukin 8, functions as a chemoattractant for polymorphonuclear leukocytes. On the other hand, an extra COOH-terminal domain of human TyrRS has cytokine activities like those of a mature human endothelial monocyte-activating polypeptide II (EMAP II). Moreover, the etiology of specific diseases (cancer, neuronal pathologies, autoimmune disorders, and disrupted metabolic conditions) is connected to specific aminoacyl-tRNA synthetases. Here we report the generation and characterization of monoclonal antibodies specific to N- and C-terminal domains of TyrRS. Recombinant TyrRS and its N- and C-terminal domains were expressed as His-tag fusion proteins in bacteria. Affinity purified proteins have been used as antigens for immunization and hybridoma cell screening. Monoclonal antibodies specific to catalytic N-terminal module and C-terminal EMAP II-like domain of TyrRS may be useful as tools in various aspects of TyrRS function and cellular localization.

Unusual domain architecture of aminoacyl tRNA synthetases and their paralogs from Leishmania major

BackgroundLeishmania major, a protozoan parasite, is the causative agent of cutaneous leishmaniasis. Due to the development of resistance against the currently available anti-leishmanial drugs, there is a growing need for specific inhibitors and novel drug targets. In this regards, aminoacyl tRNA synthetases, the linchpins of protein synthesis, have received recent attention among the kinetoplastid research community. This is the first comprehensive survey of the aminoacyl tRNA synthetases, their paralogs and other associated proteins from L. major.ResultsA total of 26 aminoacyl tRNA synthetases were identified using various computational and bioinformatics tools. Phylogenetic analysis and domain architectures of the L. major aminoacyl tRNA synthetases suggest a probable archaeal/eukaryotic origin. Presence of additional domains or N- or C-terminal extensions in 11 aminoacyl tRNA synthetases from L. major suggests possibilities such as additional tRNA binding or oligomerization or editing activity. Five freestanding editing domains were identified in L. major. Domain assignment revealed a novel asparagine tRNA synthetase paralog, asparagine synthetase A which has been so far reported from prokaryotes and archaea.ConclusionsA comprehensive bioinformatic analysis revealed 26 aminoacyl tRNA synthetases and five freestanding editing domains in L. major. Identification of two EMAP (endothelial monocyte-activating polypeptide) II-like proteins similar to human EMAP II-like proteins suggests their participation in multisynthetase complex formation. While the phylogeny of tRNA synthetases suggests a probable archaeal/eukaryotic origin, phylogeny of asparagine synthetase A strongly suggests a bacterial origin. The unique features identified in this work provide rationale for designing inhibitors against parasite aminoacyl tRNA synthetases and their paralogs.

Cigarette Smoke–Induced CXCR3 Receptor Up-Regulation Mediates Endothelial Apoptosis

Endothelial monocyte-activating polypeptide II (EMAP II) and interferon-inducible protein (IP)-10 are proinflammatory mediators, which in addition to their chemokine activities, selectively induce apoptosis in endothelial cells and are up-regulated in the lungs of cigarette smoke-exposed humans. Previously, we showed that EMAP II is an essential mediator of cigarette smoke-induced lung emphysema in mice linking endothelial cell apoptosis with inflammation. Here we addressed the role of the CXCR3 receptor in EMAP II-induced and IP-10-induced apoptosis in endothelial cells and its regulation by cigarette smoke. We found that both neutralizing antibodies and small inhibitory RNA to CXCR3 abrogated EMAP II-induced and IP-10-induced endothelial caspase-3 activation and DNA fragmentation. CXCR3 receptor surface expression in human lung microvascular endothelial cells and in lung tissue endothelium was up-regulated by exposure to cigarette smoke. In tissue culture conditions, EMAP II-induced and IP-10-induced apoptosis was enhanced by preincubation with cigarette smoke extract. Interestingly, serum starvation also induced CXCR3 up-regulation and enhanced EMAP II-induced endothelial apoptosis. Signal transduction via p38 mitogen-activated protein kinase activation was essential for CXCR3-induced cell death, but not for CXCR3 receptor up-regulation by cigarette smoke. In turn, protein nitration was required for CXCR3 receptor up-regulation by cigarette smoke and consequently for subsequent CXCR3-induced cell death. In conclusion, the concerted up-regulation of proinflammatory EMAP II, IP-10, and CXCR3 by cigarette smoke could sustain a cascade of cell death that may promote the alveolar tissue loss noted in human emphysema.

Regulation of SFRP-1 Expression in the Rat Dental Follicle

Tooth eruption requires osteoclastogenesis and subsequent bone resorption. Secreted frizzled-related protein-1 (SFRP-1) negatively regulates osteoclastogenesis. Our previous studies indicated that SFRP-1 is expressed in the rat dental follicle (DF), with reduced expression at days 3 and 9 close to the times for the major and minor bursts of osteoclastogenesis, respectively; but it remains unclear as to what molecules contribute to its reduced expression at these critical times. Thus, it was the aim of this study to determine which molecules regulate the expression of SFRP-1 in the DF. To that end, the DF cells were treated with cytokines that are maximally expressed at days 3 or 9, and SFRP-1 expression was determined. Our study indicated that colony-stimulating factor-1 (CSF-1), a molecule maximally expressed in the DF at day 3, down-regulated SFRP-1 expression. As to endothelial monocyte-activating polypeptide II (EMAP-II), a highly expressed molecule in the DF at day 3, it had no effect on the expression of SFRP-1. However, when EMAP-II was knocked down by siRNA, the expression of SFRP-1 was elevated, and this elevated SFRP-1 expression could be reduced by adding recombinant EMAP-II protein. This suggests that EMAP-II maintained a lower level of SFRP-1 in the DF. TNF-α is a molecule maximally expressed at day 9, and this study indicated that it also down-regulated the expression of SFRP-1 in the DF cells. In conclusion, CSF-1 and EMAP-II may contribute to the reduced SFRP-1 expression seen on day 3, while TNF-α may contribute to the reduced SFRP-1 expression at day 9.

1H, 13C, and 15N chemical shifts assignments for human endothelial monocyte-activating polypeptide EMAP II

Endothelial and monocyte-activating polypeptide II (EMAP II) is a cytokine that plays an important role in inflammation, apoptosis and angiogenesis processes in tumour tissues. Structurally, the EMAP II is a 169 amino acid residues long C-terminal domain (residues 147-312) of auxiliary tRNA binding protein p43. In spite of existence in pdb databank of two X-ray structures there are some important aspects of EMAP II cytokine function which are still not fully understood in detail. To obtain information about 3D structure and backbone dynamic processes in solution we perform structure evaluation of human EMAP II cytokine by NMR spectroscopy. The standard approach to sequence-specific backbone assignment using 3D NMR data sets was not successful in our studies and was supplemented by recently developed 4D NMR experiments with random sampling of evolution time space. Here we report the backbone and side chain (1)H, (13)C, and (15)N chemical shifts in solution for recombinant EMAP II cytokine together with secondary structure provided by TALOS+software.

Endothelial-monocyte activating polypeptide II disrupts alveolar epithelial type II to type I cell transdifferentiation

BackgroundDistal alveolar morphogenesis is marked by differentiation of alveolar type (AT)-II to AT-I cells that give rise to the primary site of gas exchange, the alveolar/vascular interface. Endothelial-Monocyte Activating Polypeptide (EMAP) II, an endogenous protein with anti-angiogenic properties, profoundly disrupts distal lung neovascularization and alveolar formation during lung morphogenesis, and is robustly expressed in the dysplastic alveolar regions of infants with Bronchopulmonary dysplasia. Determination as to whether EMAP II has a direct or indirect affect on ATII→ATI trans-differentiation has not been explored.MethodIn a controlled nonvascular environment, an in vitro model of ATII→ATI cell trans-differentiation was utilized to demonstrate the contribution that one vascular mediator has on distal epithelial cell differentiation.ResultsHere, we show that EMAP II significantly blocked ATII→ATI cell transdifferentiation by increasing cellular apoptosis and inhibiting expression of ATI markers. Moreover, EMAP II-treated ATII cells displayed myofibroblast characteristics, including elevated cellular proliferation, increased actin cytoskeleton stress fibers and Rho-GTPase activity, and increased nuclear:cytoplasmic volume. However, EMAP II-treated cells did not express the myofibroblast markers desmin or αSMA.ConclusionOur findings demonstrate that EMAP II interferes with ATII → ATI transdifferentiation resulting in a proliferating non-myofibroblast cell. These data identify the transdifferentiating alveolar cell as a possible target for EMAP II's induction of alveolar dysplasia.

Endothelial monocyte-activating polypeptide- II and tumor

Endothelial monocyte-activating polypeptide-Ⅱ (EMAP-Ⅱ ) is a novel proinflammatory cytokine with proinflammatory,proapoptotic and antiangiogenic properties.It is associated with many tumorassociated proteins,such as tumor necrosis factor,vascular endothelial growth factor,hypoxia inducible factor-1α,arginyl-tRNA synthetase,and insulin-like growth factor- Ⅰ.EMAP-Ⅱ has anti-tumor properties and has a good prospect in cancer prevention and treatment. Key words: Neoplasms; Neoplasm proteins; Endothelial monocyte-activating polypeptide-Ⅱ

Necessity for Re-Vascularization after Spinal Cord Injury and the Search for Potential Therapeutic Options

Disruption of the blood-spinal cord barrier (BSCB) and microvascular changes leading to reduction of blood supply represent hallmarks of spinal cord secondary injury causing further deterioration of the traumatized patient. Injury to the blood vessels starts with prominent hemorrhage and generation of inflammation. Furthermore, spinal cord ischemia and extravasation of blood components contribute to edema formation resulting in death of neural cells. Endogenous attempts of re-vascularization have been observed although these newly formed vessels display morphological and functional abnormalities. The unfavorable regulation of angiogenic and counterregulatory anti-angiogenic factors during the complicated course of vessel remodeling after SCI is suspected to participate in the failure of re-vascularization and vessel stabilization. Repression of the expression of angiogenic factors such as vascular endothelial growth factor-A (VEGF-A), placental growth factor (PlGF), angiopoietin-1 (Ang1), and platelet-derived growth factor-BB (PDGF-BB) contributes to vessel regression. Therefore, therapeutic applications of angiogenic factors following SCI are promising strategies to restore blood flow in the lesion.

Role of RhoA/ROCK Signaling in Endothelial-Monocyte-Activating Polypeptide II Opening of the Blood–Tumor Barrier

The purpose of the present study was to determine the potential for RhoA/ROCK signaling to play a role in endothelial-monocyte-activating polypeptide (EMAP) II-induced increase in blood-tumor barrier (BTB) permeability in rat brain microvascular endothelial cells (RBMECs). In the present study, we used an in vitro BTB model, a RhoA inhibitor (C3 exoenzyme) and a ROCK inhibitor (Y27632) to determine whether RhoA/ROCK pathway play a role in the process of TJ disassembly, stress fiber formation, MLC and cofilin phosphorylation, as well as increase of BTB permeability induced by EMAP II. The results revealed that BTB permeability was increased by EMAP II induction, and C3 exoenzyme or Y27632 could partially inhibit the EMAP II-induced increase of BTB permeability. The significant down-regulations in tight junction (TJ)-associated proteins occludin, claudin-5 and ZO-1 and stress fiber formation by EMAP II administration were observed, which were partly prevented by C3 exoenzyme or Y27632 pretreatment. Moreover, the significant increases in RhoA activity, myosin light chain (MLC) and cofilin phosphorylation by EMAP II administration were observed, MLC and cofilin phosphorylation were partly inhibited by C3 exoenzyme or Y27632 pretreatment. The present study demonstrates that the activation of RhoA/ROCK signaling in RBMECs was required for the increase of BTB permeability and these effects are related with the ability for RhoA/ROCK to mediate TJ disassembly and stress fiber formation by phosphorylating cofilin and MLC.

Enhancing cytotoxic agent activity in experimental pancreatic cancer through EMAP II combination therapy

Gemcitabine has limited benefits as single agent or in combination for pancreatic ductal adenocarcinoma (PDAC). Endothelial monocyte-activating polypeptide II (EMAP) enhances gemcitabine effects in PDAC. We evaluated the combination effects of EMAP, doxorubicin, and docetaxel in experimental PDAC. Cell proliferation, protein expression, and apoptosis were analyzed by WST-1 assay, Western blotting, and FACS analysis. Tumor growth and survival experiments were performed in murine xenografts. PDAC cell proliferation in vitro was not affected by EMAP, compared to a small inhibition through doxorubicin, docetaxel, and gemcitabine. EMAP addition to these agents did not increase the antiproliferative effects. In endothelial cells, EMAP, doxorubicin, docetaxel, and gemcitabine all had antiproliferative effects. Addition of EMAP to these cytotoxic agents had additive effects. In PDAC cells, no agent induced measurable apoptosis, whereas in endothelial cells, all agents either alone or in combination did. Doxorubicin, docetaxel, gemcitabine, and EMAP all decreased tumor growth. EMAP addition increased inhibitory effects of docetaxel and gemcitabine, but not of doxorubicin. However, compared to controls (median survival: 17days), EMAP (14days) had no survival benefit, while docetaxel (29days), gemcitabine (25days), and docetaxel followed by gemcitabine sequence (37days) extended animal survival. Addition of EMAP to docetaxel (35days), gemcitabine (28days), and docetaxel gemcitabine sequence (41days) extended the survival. Doxorubicin effects were not enhanced by EMAP. The antiendothelial combination therapy benefit through EMAP is not limited to gemcitabine and may facilitate the development of more effective alternative cytotoxic therapy strategies against PDAC.

Lesional Expression of EMAPII in Macrophages/Microglia Following Cerebral Ischemia in Rats

ABSRACTObjective: Cerebral ischemia triggers acute inflammation, which exacerbates primary brain damage. Characterization of cytokine expression to the early damaged tissue might aid in further understanding of lesion development and contribute to definition of molecular targets for selective immunotherapy. Endothelial monocyte-activating polypeptide II (EMAPII) is a proinflammatory, antiangiogenic cytokine whose expression following cerebral ischemia remained unknown. Therefore, we studied the spatiotemporal expression of EMAPII in early brain lesions after cerebral ischemia–reperfusion injury. Methods: Unilateral transient focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 1 hr followed by different reperfusion periods using male Sprague-Dawley rats. Subsequently, rats were sacrificed on Day 1, 3, 5, or 7 following surgery. EMAPII expression was investigated by immunohistochemistry. Results: EMAPII-positive cell accumulation was observed as early as Day 1 postreperfusion and increased steadily. Significant accumulation of EMAPII-positive cells was seen in lesion and penumbra areas but not in the translateral hemisphere. Both amoeboid and ramified EMAPII-positive cells were observed and mainly localized to lesion and penumbra areas, respectively. Conclusion: The known pathological functions together with abundant cellular accumulation in cerebral ischemia lesions suggest that EMAPII might contribute to pathophysiological consequences of cerebral ischemia.