The BGJ398 mouse grades with less than 2+ were considered as low reactivity for VEGF, otherwise as high reactivity. Evaluation of microvessel density Microvessels were identified by immunostaining endothelial cells with the mouse anti-human

monoclonal antibody CD34. Microvessel density (MVD) was assessed according to the international consensus [12]. The entire section was scanned systematically at low magnification (× 100) in order to identify the most intense areas of neovascularization (“”hotspots”") within the tumor. After five hotspots areas with the highest number of capillaries and small venules were identified, microvessels were counted at high power magnification (× 400), and the average of count in five fields was calculated. MVD was quoted as a continuous variable [13, 14]. Statistical analysis The selleck chemicals Chi-square test or Fisher’s exact probability test for proportion was used to analyze the relationship between SPARC and VEGF expression, and clinicopathologic characteristics. One-way ANOVA test and Linear regression analysis was used to assess the correlations among the continuous variables. Spearman rank correlation coefficient test analysis was performed to examine the correlations among different variables.. Survival curves were plotted by the Kaplan-Meier method, and compared by the log-rank test. To identify independent prognostic factors, including cancer recurrence,

distant metastasis or death from disease, the Cox regression analysis was performed with the endpoints for disease-free survival (DFS) and overall survival (OS), respectively. A P-value of less than 0.05 was considered statistically significance. SPSS 11.5 was used for the statistical analysis. Results Expression of SPARC, VEGF, and CD34 in colon cancer and normal colon mucosa tissue Expression of SPARC protein was determined by immunohistochemistry staining in 114 cases of paraffin-embedded colon cancer tissues and their corresponding non-diseased colon tissue. SPARC was mainly localized in the cytoplasm and was detected in the normal colonic epithelial cells (Fig 1a), the colon

cancer cells and the mesenchymal and stromal Methocarbamol cells (MSC) of colon cancer (Fig 1b). In this study, the degree of the expression of SPARC showed that 81 cases (71.1%) with low reactivity and 33 cases (28.9%) with high reactivity in tumor cells, 61 cases (53.5%) with low reactivity and 53 cases (46.5%) with high reactivity in the MSC surrounding the tumor, and 84 cases (73.7%) with low reactivity and 30 cases (26.3%) with high reactivity in the normal colon mucosa tissue, respectively. SPARC expression was no significant difference between the reactivity in tumor cells and in their corresponding non-diseased colon mucosa (P > 0.05), but was statistically significant difference between that in MSC and in tumor cells (P < 0.05), and between that in MSC and normal mucosa in colon tissue (P < 0.05), respectively.

The mouse anti-EfTu antibody was a kind gift from Dr. YX Zhang, Boston, USA. Rat anti-HA antibody was from Roche and the TRITC-conjugated

anti-rat antibody was from Jackson Immuno Research. Cy™-5-conjugated goat anti-mouse antibody was purchased from Amersham. INPs Two SCH772984 salicylidene acylhydrazides, namely INP0400 and INP0341, were provided by Innate Pharmaceuticals AB, Umeå, Sweden. The compounds were dissolved in dimethyl sulfoxide (DMSO, Sigma) as 10 mM stock solutions and used at the concentrations indicated. Chlamydia entry assay HeLa cells were infected with C. trachomatis L2 or C. caviae GPIC in the presence or absence of 60 μM INP0400 or INP0341 and centrifuged for 5 minutes at 770 g at room temperature. Cells were fixed 2.5 h later and extracellular and intracellular bacteria were labelled as described [11]. In brief, extracellular bacteria were labelled with anti-Chlamydia antibody followed by anti-mouse Cy™-5 antibody. The cells were then permeabilized in

PBS containing 0.05% saponin and 1 mg/ml BSA and intracellular bacteria were labelled with FITC-conjugated anti-Chlamydia antibody. The number of extracellular and intracellular bacteria was counted in 15 fields, with an average of 75 bacteria per field, in two independent experiments. The efficiency of entry is expressed as the ratio of intracellular to total cell-associated bacteria (intracellular and extracellular). Immunofluorescence selleck antibody microcopy To visualize the effect of the drugs on Chlamydia development, HeLa cells infected with C. trachomatis L2 or C. caviae GPIC were grown in presence of INPs (or DMSO for control) for 24 h, fixed, and labelled with anti-EfTu antibody followed by Alexa488-coupled

goat anti-mouse antibody. DNA was stained with 0.5 μg/ml Hoechst 33342 in the mounting medium. Recruitment of actin to bacterial entry sites was visualized with Alexa546-phalloidin in HeLa cells infected with FITC-labelled C. caviae in the presence or absence of 60 μM INP0341 as described [11]. To visualize Arf6 and Rac distribution, Arachidonate 15-lipoxygenase cells were transfected with HA-tagged Arf6 or GFP-tagged Rac. Hela cells were infected with C. caviae GPIC 24 h after transfection and spun for 5 minutes at 770 g at room temperature. At 10 minutes p.i. cells were fixed and labelled with Alexa546-phalloidin (GFP-Rac transfected cells) or Alexa488-phalloidin (Arf6-HA transfected cells). Arf6 was labelled with a rat anti-HA antibody (Roche, clone 3F10) followed by a TRITC-conjugated anti-rat antibody (Jackson Immuno Research). Immunofluorescence microcopy was performed with an epifluorescence microscope (Axiophot, Zeiss, Germany) attached to a cooled CDD camera (Photometrics, Tucson, AZ), using a 63× Apochromat lens. Acknowledgements This work was supported by the European Marie Curie program European Initiative for basic research in Microbiology and Infectious Diseases and by the Agence Nationale pour la Recherche (ANR-06-JCJC-0105).

Biomark Med 2013, 7:779–790.PubMedCrossRef 43. Szeto CC: Urine miRNA in nephrotic syndrome. Clin Chim Acta 2014, 436C:308–313.PubMedCrossRef Competing interests The authors declare no competing financial interests. Authors’ contributions Y-GF conceived the project; designed the experiments and carried out the majority

of the experiments; JL conducted the bioinformatics analysis; Y-MK, YH and BL helped to collect clinical samples. PY and ZY helped to culture cells; all authors discussed the results; Y-GF and JL wrote the manuscript. All authors read and approved the final manuscript.”
“Background Gastric cancer (GC) is the second leading cause of global cancer mortality, accounting for 700,000 deaths annually [1,2]. More than 70% of countries worldwide have learn more a mortality-to-incidence ratio greater than 0.8, suggesting that prevention of late presentation and modified Selinexor in vivo treatment strategies are required to improve clinical outcomes [3]. In particular, distant metastases including peritoneal dissemination have been recognized as important prognostic determinants for GC patients [4,5]. Identifying genes relevant to the malignant behavior of GC could aid clinicians in tailoring treatments by identifying high-risk patients and proposing novel molecular targets [6]. Recently, technological advances such as microarrays and next-generation sequencing have allowed for the exhaustive

genomic characterization of malignancies, enhancing our understanding of cancer initiation and progression [7-9]. With these techniques, numerous genetic and epigenetic alterations relevant to gastric carcinogenesis and GC progression have been reported [10].

However, understanding the clinical significance of individual genes remains insufficient, despite the accumulating array data. Dihydropyrimidinase-like 3 (DPYSL3) is a cell-adhesion molecule [11,12] and actively expressed in normal tissues of cardiac myocytes, brain, pineal body, retina and smooth muscle, and moderately expressed in Protein kinase N1 various tissues including gastric tissues [13]. DPYSL3 has been reported to be involved in the metastatic process of tumor cells [14,15]. Gao et al. conducted expression and functional analyses of DPYSL3 in prostate cancer and found that DPYSL3 is a metastasis suppressor that is inversely associated with the expression of vascular endothelial growth factor (VEGF) [14]. In contrast, Kawahara et al. reported that DPYSL3 facilitates pancreatic cancer cell metastasis via a strong interaction with other cell adhesion factors, including ezrin (EZR), focal adhesion kinase (FAK) and c-SRC [15]. Thus, DPYSL3 has attracted attention as a metastatic modulator; however, the role of DPYSL3 expression in GC initiation and progression has not been investigated. Here, we focused on DPYSL3 as a potential facilitator of malignant behavior in GC. The aim of this study was to evaluate the clinical significance of DPYSL3 expression in GC.

Ecological implications of anaerobic

nitrate turnover by isolate An-4 Aspergillus terreus is a common and globally occurring soil fungus that is also known from substrates as diverse as air, salterns, capybara droppings, lung of pocket mice, corn, cotton plants, milled rice, muesli, and wall paint [39]. The species has been reported from marine and associated habitats, such as mangroves and soft corals, and isolates from these habitats have been widely investigated for the production of bioactive compounds [40–42]. A. terreus has also been isolated from the hypersaline water of the Dead Sea [43, 44]. The species is an important human pathogen causing bronchopulmonary aspergillosis and disseminated infections [45]. Dissimilatory NO3 – reduction by human-associated microorganisms has been demonstrated [46, 47], but it is not known whether fungi are selleck chemicals involved. A. terreus is also of considerable biotechnological interest because it produces a wide diversity

of secondary metabolites that find pharmaceutical applications, biotechnologically Adriamycin clinical trial relevant compounds such as itaconic acid and itatartaric acid, as well as mycotoxins that are important for food safety ( [39] and references therein). The wide habitat spectrum of A. terreus might be significantly expanded by the ability for dissimilatory NO3 – reduction in the absence of O2. This fungus has the potential to survive hypoxic or anoxic conditions that prevail in aquatic sediments mostly just a few millimeters below the surface [48] or even

directly at the surface when O2 concentrations are low in the water column [12, 49]. In contrast, NO3 – originating from the water column and/or the nitrification layer at the sediment surface diffuses deeper into the sediment than O2 does [50]. In shallow sediments, NO3 –rich water is introduced into even deeper layers by mixing forces such as bioturbation, bioirrigation, and ripple movement [51, 52]. The sediment habitat in which A. terreus can thrive is further expanded by its NO3 – storage capability. The maximum intracellular NO3 – content of 8 μmol g-1 protein theoretically sustains dissimilatory NO3 – reduction without extracellular NO3 – supply for 2–4.5 days (calculated from rates measured in the 15N-labeling experiment). Survival and mTOR inhibitor growth beyond this time frame will depend on the ability of A. terreus to repeatedly access NO3 – in its natural sediment habitat, which is currently unknown. The dissimilatory NO3 – reduction activity of An-4 leads to the production and release of NH4 +, N2O, and NO2 -. Thus, unlike the denitrification and anammox activities of other microorganisms, the anaerobic NO3 – metabolism of An-4 cannot directly lead to fixed nitrogen removal. Since the major product of NO3 – reduction is NH4 +, An-4 merely converts one form of fixed nitrogen into another one.

McClung JP, Karl JP, Cable SJ, STAT inhibitor Williams KW, Young AJ, Lieberman HR: Longitudinal decrements in iron status during military training

in female soldiers. Br J Nutr 2009, 102:605–609.CrossRefPubMed 5. Ruohola JP, Laaksi I, Ylikomi T, Haataja R, Mattila VM, Sahi T, Tuohimaa P, Pihlajamaki H: Association between serum 25(OH)D concentrations and bone stress fractures in Finnish young men. J Bone Miner Res 2006, 21:1483–1488.CrossRefPubMed 6. Jones BH, Thacker SB, Gilchrist J, Kimsey CD Jr, Sosin DM: Prevention of lower extremity stress fractures in athletes and soldiers: a systematic review. Epidemiol Rev 2002, 24:228–247.CrossRefPubMed 7. Friedl KE, Evans RK, Moran DS: Stress fracture and military medical readiness: bridging basic and applied beta-catenin mutation research. Med Sci Sports Exerc 2008,40(Suppl 11):S609-S622.PubMed 8. Vieth R, Cole DE, Hawker GA, et al.: Wintertime vitamin D insufficiency is common in young Canadian women, and their vitamin D intake does not prevent it. Eur J Clin Nutr 2001, 55:1091–1097.CrossRefPubMed 9. Harris SS: Vitamin D and African Americans. J Nutr 2006, 136:1126–1129.PubMed 10. Karl JP, Lieberman HR, Cable SJ, Williams KW, Glickman EL, Young AJ, McClung JP: Poor iron status is not associated with overweight or overfat in non-obese pre-menopausal women.

J Am Coll Nutr 2009, 28:37–42.PubMed 11. McClung JP, Karl JP, Cable SJ, Williams KW, Nindl BC, Young AJ, Lieberman HR: Randomized, double-blind, placebo-controlled trial of iron supplementation in female soldiers during military training: effects on iron status, physical performance, and mood. Am J Clin Nutr 2009, 90:1–8.CrossRef 12. Knapik JJ, Darakjy

S, Hauret KG, Canada S, Marin R, Jones BH: Ambulatory physical activity during United States Army Basic Combat Training. Int J Sports Med 2007, 28:106–115.CrossRefPubMed 13. Vieth Org 27569 R, Bischoff-Ferrari, Boucher BJ, Dawson-Hughes B, Garland CF, Heaney RP, Holick MF, Hollis BW, Lamberg-Allardt C, McGrath JJ, Norman AW, Scragg R, Whiting SJ, Willett WC, Zittermann A: The urgent need to recommend an intake of vitamin D that is effective. Am J Clin Nutr 2007, 85:649–650.PubMed 14. Dawson-Hughes B, Heaney RP, Holick MF, Lips P, Meunier PJ, Vieth R: Estimates of optimal vitamin D status. Osteoporos Int 2005, 16:713–716.CrossRefPubMed 15. Looker AC, Dawson-Hughes B, Calvo MS, Gunter EW, Sahyoun NR: Serum 25-hydroxyvitamin D status of adolescents and adults in two seasonal subpopulations from NHANES III. Bone 2002, 30:771–777.CrossRefPubMed 16. Nesby-O’Dell S, Scanlon KS, Cogswell ME: Hypovitaminosis D prevalence and determinants among African American and white women of reproductive age: third National Health and Nutrition Examination Survey, 1988–1994. Am J Clin Nutr 2002, 76:187–192.PubMed 17. Dawson-Hughes B: Racial/ethnic considerations in making recommendations for vitamin D for adult and elderly men and women. Am J Clin Nutr 2004,80(Suppl 6):S1763-S1766. 18.

Respondents were similar to non-respondents in terms of fracture history, osteoporosis diagnosis, and osteoporosis treatment [9], as determined by self-reported data collected at baseline [10]. Data sources and measures Inhibitor Library supplier Study questionnaire

(self-report of drug use and DXA testing) As part of the standardized telephone interview completed in 2003/2004, we asked participants if they had ever had a bone density test and recorded information regarding osteoporosis pharmacotherapy (bisphosphonates, calcitonin, and raloxifene) and the use of other agents that may impact bone density (estrogen therapy, glucocorticoids, and thyroid medication) as current, past, or never. Question wording is included in the “Appendix.” DXA confirmation and DXA—documented osteoporosis DXA results were sought from participants who reported having had a DXA test and who completed a signed release of information form. For these patients, physicians were contacted to confirm that a DXA was completed and to obtain a copy of the most recent DXA report. We previously reported that the positive predictive value for self-report of having had a DXA was 93% when using physician responses as the gold standard [5]. Among those with a DXA report, we categorized

bone mineral density according to the lowest T-score at the lumbar spine (L1-4 or L2-4) or hip (femoral neck or total hip) as normal (T-score ≥ −1), osteopenic (−1 < T-score > −2.5), or osteoporotic (T-score ≤ −2.5) [11]. Healthcare utilization data—medical claims In Canada, physician and hospital services are funded through publicly financed comprehensive universal

health insurance. BIBW2992 In Ontario, claims for physician services are documented in the Ontario Health Insurance Plan (OHIP) Claims History Database. Information about inpatient services are captured in the Canadian Institutes of Health Information Discharge Abstract Database, and information about emergency department services are documented in the National Ambulatory Care Reporting System. Prior to April 1, 2002, hospital and emergency department records were coded using the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM). Mirabegron Since then, they have been coded using ICD-10-Canada (CA). July 1991 is the earliest date for which individual level data are available. DXA tests were identified using OHIP claim codes: J654, J655, J656, J688, J854, J855, J856, J888, X145, X146, X149, X152, X153, X155, and X157. These include codes for dual-photon absorptiometry, which predates DXA technology and was used prior to April 1998 [12]. We considered claims back to July 1991 when individual level claims data were first recorded in Ontario. Osteoporosis diagnosis was identified by any OHIP diagnosis code of 733 or any hospitalization or emergency department visit code of ICD-9-CM = 733.0 or ICD-10-CA = M80, M81, or M82. We considered diagnosis within 1 year pre- and post-DXA, as well as within 1 to 5 years before questionnaire completion.

World J Surg 2004, 28:301–306.CrossRef 7. Wain J, Diep TS, Ho VA, Walsh AM, Hoa NTT, Parry CM: Quantitation of bacteria in blood of typhoid fever patients and relationship between counts and clinical features, transmissibility, and antibiotic resistance. J Clin Microbiol 1998, 36:1683–1687. 8. Stewart PS, Costerton JW: Antibiotic resistance of bacteria in biofilms. Lancet 2001, 358:135–138.CrossRef 9. Hetrick EM, Shin JH, Stasko NA, Johnson CB, Wespe DA, Holmuhamedov E, Schoenfisch MH: Bactericidal efficacy of

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CC, McMullin B, Ghaharya A: Potential application of gaseous nitric oxide as a topical antimicrobial agent. Nitric Oxide 2006, 14:21–29.CrossRef 20. Marxer SM, Rothrock AR, Nablo BJ, Robbins ME, Schoenfisch MH: Preparation of nitric oxide (NO)-releasing sol - gels for biomaterial applications. Chem Mater 2003, 15:4193–4199.CrossRef 21. Carpenter AW, Slomberg DL, Rao KS, Schoenfisch MH: Influence CYTH4 of scaffold size on bactericidal activity of nitric oxide-releasing silica nanoparticles. ACS Nano 2011, 5:7235–7244.CrossRef 22. Hetrick EM, Shin JH, Paul HS, Schoenfisch MH: Anti-biofilm efficacy of nitric oxide-releasing silica nanoparticles. Biomaterials 2009, 30:2782–2789.CrossRef 23. Friedman AJ, Han G, Navati MS, Chacko M, Gunther L, Alfieri A, Friedman JM: Sustained release nitric oxide releasing nanoparticles: characterization of a novel delivery platform based on nitrite containing hydrogel/glass composites. Nitric Oxide 2008, 19:12–20.CrossRef 24.

The cytokine encoded by this gene may also play a role in mediating homing of lymphocytes to secondary lymphoid organs. CSF3 (granulocytes colony stimulation factor 3) is a cytokine that controls the production, differentiation, and function of granulocytes. We may speculate

that the specific expression of the last two genes might contribute to severity of the inflammation at later stages of infection as caused by this pathogen in vivo. Conclusion We employed DNA expression microarrays to study the early transcriptional response of naïve human peripheral monocytes infected with a set of three important gram-positive bacterial pathogens: Staphylococcus aureus, Streptococcus pneumoniae and Listeria monocytogenes. Upregulation of chemokine rather Neratinib than interleukin genes was characteristic for the early response with the exception of the prominent expression of IL23, marking it as the lead early cytokine. An important finding was the observed activation of genes regulating angiogenesis and endothelial cell function together with genes involved in managing pathogen induced cytoplasmic stress and counteracting apoptosis. This transcription program seems to be characteristic for the first events in monocyte activation and points to induction of cytokine

signalling rather than to a program change of naïve monocytes to pathogen eliminating effector cells. Methods Isolation of CD14 positive WBCs from human peripheral blood Blood

concentrates (buffy coats) were obtained routinely at Wnt inhibitor Dapagliflozin the transfusion center, clinic of JLU Gießen. Approval for the use of clinical material in this study was in compliance with procedures laid down by the Helsinki Declaration and approved by the Ethics Study Board of the University Hospital of Giessen (File number 79/01). For the isolation of monocytes, only fresh (1 to 1.5 hour old) buffy coats from phenotypic healthy donors (3 males + 2 females) were used. The isolation of the mononuclear leucocytes was done by centrifugation trough a ficol cushion (Ficol-Plaque-TM, Amersham Biosciences). After the centrifugation the interphase was collected and the cells were washed twice with PBS. The cells were reconstituted in PBS and kept on ice. Anti-CD14 antibody labeled magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany) were added to the cells in a ratio of 20 μl/107 cells (ca. 5 Abs./cell). After 15 min. incubation at 4°C unbound beads were separated by a short centrifugation step and the labeled cells were loaded and purified on a LS positive selection column using the MidiMACS magnetic separator (Miltenyi Biotec, Bergisch Gladbach, Germany) following the manufacturers instruction. The CD14+ cells were eluted in PBS and an aliquot was used for cell counting.

Ann Surg Oncol 2007, 14:258–269.PubMedCrossRef 7. Petrowsky H, Roberts GD, Kooby DA, Burt BM, Bennett JJ, Delman KA, Stanziale SF, Delohery HM781-36B cell line TM, Tong WP, Federoff HJ, Fong Y: Functional interaction

between fluorodeoxyuridine-induced cellular alterations and replication of a ribonucleotide reductase-negative herpes simplex virus. J Virol 2001, 75:7050–7058.PubMedCentralPubMedCrossRef 8. Cunningham D, Allum WH, Stenning SP, Thompson JN, Van de Velde CJ, Nicolson M, Scarffe JH, Lofts FJ, Falk SJ, Iveson TJ, et al.: Perioperative chemotherapy versus surgery alone for resectable gastroesophageal cancer. N Engl J Med 2006, 355:11–20.PubMedCrossRef 9. Vaha-Koskela MJ, Heikkila JE, Hinkkanen AE: Oncolytic viruses Cisplatin concentration in cancer therapy. Cancer Lett 2007, 254:178–216.PubMedCrossRef 10. Chen N, Zhang Q, Yu YA, Stritzker J, Brader P, Schirbel A, Samnick S, Serganova I, Blasberg R, Fong Y, Szalay AA: A novel recombinant vaccinia virus expressing the human norepinephrine transporter retains oncolytic potential and facilitates deep-tissue imaging. Mol Med 2009, 15:144–151.PubMedCentralPubMedCrossRef 11. Zhang Q, Yu YA, Wang E, Chen N, Danner RL, Munson PJ, Marincola FM, Szalay AA: Eradication of solid human breast

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Brader P, Kelly KJ, Chen N, Yu YA, Zhang Q, Zanzonico P, Burnazi EM, Ghani RE, Serganova I, Hricak H, et al.: Imaging a Genetically Engineered Oncolytic Vaccinia Virus (GLV-1 h99) Using a Human Norepinephrine Transporter Reporter Gene. Clin Cancer Res 2009, 15:3791–3801.PubMedCrossRef 14. Crew KD, Neugut AI: Epidemiology of gastric cancer. World J Gastroenterol 2006, 12:354–362.PubMed 15. Yamada E, Miyaishi S, Nakazato H, Kato K, Kito T, Takagi H, Yasue M, Kato T, Morimoto much T, Yamauchi M: The surgical treatment of cancer of the stomach. Int Surg 1980, 65:387–399.PubMed 16. Khan FA, Shukla AN: Pathogenesis and treatment of gastric carcinoma: “”an up-date with brief review”". J Cancer Res Ther 2006, 2:196–199.PubMedCrossRef 17. Liu TC, Kirn D: Gene therapy progress and prospects cancer: oncolytic viruses. Gene Ther 2008, 15:877–884.PubMedCrossRef 18. Shen Y, Nemunaitis J: Fighting cancer with vaccinia virus: teaching new tricks to an old dog. Mol Ther 2005, 11:180–195.PubMedCrossRef 19. B M: Poxviridae: the Viruses and Their Replication. 4th edition. Philadelphia: Lippincort Williams & Wilkins; 2001. 20.

1H NMR (DMSO-d 6) δ (ppm): 7.60 (t, 3H, CHarom., J = 3.6 Hz), 7.56–7.55 (m, 1H, CHarom.), 7.53–7.48 (m, 2H, CHarom.), 7.47–7.44 (m, 6H, CHarom.), 7.40–7.31 (m, 3H, CHarom.), 7.20–7.08 (m, 2H, CHarom.), 6.23 (d, 1H, CHarom., J = 7.8 Hz), 3.51–3.28 (m, 6H, CH2), 3.19–3.07 (m, 6H, CH2), 1.70–1.68

(m, 2H, CH2), 1.58–1.53 (m, 6H, CH2). 13C NMR (CDCl3) δ (ppm): 190.30, 165.71, 165.49, 149.83, 148.79, 141.26, 137.44, 135.86, 134.92, 134.77, 134.51, 133.34 (2C), 132.58 (2C), 130.93 (2C), 129.81 (2C), 129.79 (2C), 128.73 (3C), 128.52 (3C), 128.39 (2C), 127.04 (2C), 124.82, 123.17, 58.14, 58.07, 52.58, 52.47, 35.97, 34.06, 29.74, 26.11. ESI MS: m/z = 652.4 [M+H]+ (100 %). Synthesis of 2-4-[4-(2-metoxyphenyl)piperazin-1-yl]butyl-4,10-diphenyl-1H,2H,3H,5H-indeno[1,2-f]isoindole-1,3,5-trione Fer-1 (19) Yield: 79 %, m.p. 245–246 °C. 1H NMR (DMSO-d 6) δ (ppm): 7.61 (t, 3H, CHarom., J = 3.6 Hz), 7.56–7.44 (m, 8H, CHarom.), 7.41–7.31 (m, 2H, CHarom.), 7.05–6.87 (m, 4H, CHarom.), 6.23 (d, 1H, CHarom., J = 6.9 Hz), 3.79 (s, 3H, OCH3), 3.47–3.44 Idasanutlin research buy (m, 6H, CH2), 3.07–2.97 (m, 6H, CH2), 1.69–1.67 (m, 2H, CH2), 1.59–1.52 (m, 2H, CH2). 13C NMR (CDCl3) δ (ppm):

192.35, 165.07, 164.79, 149.81, 148.96, 141.13, 137.77, 135.42, 134.37, 134.26, 134.08, 133.11 (2C), 132.66 (2C), 130.72 (3C), 129.86, 129.72 (2C), 128.91 (3C), 128.54 (2C), 128.21 (3C), 127.75 (2C), 124.11, 123.59, 62.00, 58.84, 58.71, 52.97, 52.84, 35.06, 34.26, 29.59, 26.91. ESI MS: m/z = 648.3 [M+H]+ (100 %). 3-4-[4-(2-Metoxyphenyl)piperazin-1-yl]butyl3-azatricyclo[7.3.1.05,13]trideca-(12),5,7,9(13),10-pentaene-2,4-dione (20) was obtained according to method presented previously (Hackling et al., 2003) Yield: 63 %, m.p. 279–282 °C. 1H NMR (DMSO-d 6) δ (ppm): 8.59–8.48 (d, 2H, CHarom., J = 8.1 Hz), 8.11 (d, 2H, CHarom., J = 7.8 Hz), 7.64 (t, 2H, CHarom., J = 7.6 Hz), 7.08–6.76 (m, 4H, CHarom.) 4.56–4.17 (m, STK38 2H, CH2), 3.87 (s, 3H, OCH3), 3,41–2.98 (m, 5H, CH2), 2.93–2.32

(m, 5H, CH2), 2.04–1.42 (m, 4H, CH2). 13C NMR (CDCl3) δ (ppm): 165.72, 159.08, 158.97, 140.62, 134.22, 134.17, 134.09, 133.74, 132.25, 130.14, 129.64, 129.53, 128.47, 128.38, 128.09, 127.48, 124.02, 123.61, 61.13, 60.95, 57.53, 51.27, 51.13, 41.37, 41.29, 26.96, 26.87. ESI MS: m/z = 344.6 [M+H]+ (100 %). Biological assays Cell-based assays Cell-based assays were performed at Dipartimento di Scienze e Tecnologie Biomediche, Università di Cagliari, Monserrato, Italy. Test compounds Compounds were dissolved in DMSO at 100 mM and then diluted in culture medium. Cells and viruses Cell line and viruses were purchased from the American Type Culture Collection (ATCC).