http://hdl.handle.net/10027/7428 Fri, 24 May 2013 00:30:03 GMT 2013-05-24T00:30:03Z http://hdl.handle.net/10027/8554 Interaction of an anticancer peptide fragment of azurin with p53 and its isolated domains studied by atomic force spectroscopy Bizzarri, Anna Rita; Santini, Simona; Coppari, Emilia; Bucciantini, Monica; Di Agostino, Silvia; Yamada, Tohru; Beattie, Craig W; Cannistraro, Salvatore p28 is a 28-amino acid peptide fragment of the cupredoxin azurin derived from Pseudomonas aeruginosa that preferentially penetrates cancerous cells and arrests their proliferation in vitro and in vivo. Its antitumor activity reportedly arises from post-translational stabilization of the tumor suppressor p53 normally downregulated by the binding of several ubiquitin ligases. This would require p28 to specifically bind to p53 to inhibit specific ligases from initiating proteosome-mediated degradation. In this study, atomic force spectroscopy, a nanotechnological approach, was used to investigate the interaction of p28 with full-length p53 and its isolated domains at the single molecule level. Analysis of the unbinding forces and the dissociation rate constant suggest that p28 forms a stable complex with the DNA-binding domain of p53, inhibiting the binding of ubiquitin ligases other than Mdm2 to reduce proteasomal degradation of p53. This is a copy of an article published in the International Journal of Nanomedicine © 2011 Bizzarri et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited. DOI: 10.2147/IJN.S26155 Wed, 23 Nov 2011 06:00:00 GMT http://hdl.handle.net/10027/8554 2011-11-23T06:00:00Z http://hdl.handle.net/10027/8525 Knockdown of MADD and c-FLIP overcomes resistance to TRAIL-induced apoptosis in ovarian cancer cells Li, Liang-Cheng; Jayaram, Shankar; Ganesh, Lakshmy; Qian, Lixia; JacobRotmensch, Jacob; Maker, Ajay V.; Prabhakar, Bellur S. Objective The clinical utility of tumor necrosis factor–related apoptosis–inducing ligand (TRAIL) in the treatment of established human malignancies is limited by the development of resistance to TRAIL. We hypothesized that knockdown of map-kinase activating death domain containing protein (MADD), a TRAIL-resistance factor, may overcome TRAIL resistance in ovarian cancer cells. Study Design MADD expression in resected ovarian cancer specimens and cell lines was quantified with the use of polymerase chain reaction. Sensitivity of ovarian cancer cell lines to TRAIL, with or without MADD knockdown, was assessed. Results MADD is expressed at relatively higher levels in human malignant ovarian cancer tissues and cell lines, compared with normal ovarian tissues. The cell lines OVCA429 and OVCAR3 were susceptible, and cell lines CAOV-3 and SKOV-3 were resistant to TRAIL. MADD knockdown in CAOV-3 cells, but not in SKOV-3 cells, conferred TRAIL sensitivity. Knockdown of cellular Fas-associated death domain–like interleukin-1 beta-converting enzyme–inhibitory protein (c-FLIP) in SKOV-3 cells increased spontaneous and TRAIL-induced apoptosis, which was further increased on MADD knockdown. Conclusion MADD/c-FLIPL knockdown can render TRAIL-resistant ovarian cancer cells susceptible to TRAIL. NOTICE: this is the author’s version of a work that was accepted for publication in American Journal of Obstetrics and Gynecology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in American Journal of Obstetrics and Gynecology, [Vol 205, Issue 4, October 2011] DOI: 10.1016/j.ajog.2011.05.035. The original publication is available at www.elsevier.com. Sat, 01 Oct 2011 05:00:00 GMT http://hdl.handle.net/10027/8525 2011-10-01T05:00:00Z http://hdl.handle.net/10027/8443 Chromatin remodeling resets the immune system to protect against autoimmune diabetes in mice Patel, Tejas; Patel, Vasu; Singh, Rajvir; Jayaraman, Sundararajan Epigenetic alteration of the genome has been shown to provide palliative effects in mouse models of certain human autoimmune diseases. We have investigated whether chromatin remodeling could provide protection against autoimmune diabetes in NOD mice. Treatment of female mice during the transition from prediabetic to diabetic stage (18-24 weeks of age) with the well-characterized histone deacetylase inhibitor, Trichostatin A effectively reduced the incidence of diabetes. However, similar treatment of overtly diabetic mice during the same time period failed to reverse the disease. Protection against diabetes was accompanied by histone hyperacetylation in pancreas and spleen, enhanced frequency of CD4+ CD62L+ cells in the spleen, reduction in cellular infiltration of islets, restoration of normoglycemia and glucose-induced insulin release by beta cells. Activation of splenic T lymphocytes derived from protected mice in vitro with pharmacological agents that bypass the antigen receptor or immobilized anti-CD3 antibody resulted in enhanced expression of Ifng mRNA and protein without altering the expression of Il4, Il17, Il18, Inos, and Tnfa genes nor the secretion of IL-2, IL-4, IL-17, and TNF-α proteins. Consistently, expression of the transcription factor involved in Ifng transcription, Tbet/Tbx21 but not Gata3 and Rorgt, respectively required for the transcription of Il4 and Il17, was upregulated in activated splenocytes of protected mice. These results indicate that chromatin remodeling can lead to amelioration of diabetes by employing multiple mechanisms including differential gene transcription. Thus, epigenetic modulation could be a novel therapeutic approach to block the transition from benign to frank diabetes. Post print version of article may differ from published version. The definitive version is available through Nature Publishing Group at DOI:10.1038/icb.2010.144 link:http://www.nature.com/icb/journal/v89/n5/full/icb2010144a.html Fri, 01 Jul 2011 05:00:00 GMT http://hdl.handle.net/10027/8443 2011-07-01T05:00:00Z http://hdl.handle.net/10027/8387 Robotic Roux-en-Y Duodenojejunostomy for Superior Mesenteric Artery Syndrome: Operative Technique Ayloo, Subhashini M.; Masrur, Mario A.; Bianco, Francesco M.; Giulianotti, Pier C. Background: Superior mesenteric artery (SMA) syndrome, also known as Wilkie's syndrome, is a rare condition characterized by vascular compression of the duodenum that leads to intestinal obstruction. While there have been a few recent case reports of laparoscopic duodenojejunostomy performed as an option for surgical treatment, the role of the da Vinci® robot in superior mesenteric syndrome has been underestimated. The authors report a robotic Roux-en-Y duodenojejunostomy for the treatment of SMA syndrome. Materials and Methods: A 39-year-old man with a history of Amyotrophic lateral sclerosis presented with an upper gastrointestinal obstruction with distended abdomen. A computed tomography scan showed a transition in the third portion of the duodenum where the SMA vessels crossed over, with a decompressed jejunum. He was identified as a candidate for a duodenojejunostomy. The da Vinci Surgical System was used to mobilize the colon and duodenum, and a Roux-en-Y duodenojejunostomy was performed with hand-sewn anastomosis. Results: There were no intraoperative complications. The blood loss was minimal and operative time was 120 minutes. The postoperative course was uneventful with resolution of intestinal obstruction. Conclusion: Robotic Roux-en-Y duodenojejunostomy as a surgical option for treatment of SMA syndrome is safe, feasible, and a valid alternative to open surgery with the added benefits of a minimally invasive approach. This is a copy of an article published in the Journal of Laparoendoscopic and Advanced Surgical Techniques © 2011. Copyright Mary Ann Liebert, Inc.; Journal of Laparoendoscopic and Advanced Surgical Techniques is available online at: http://www.liebertonline.com. DOI: 10.1089/lap.2011.0070 Fri, 05 Aug 2011 05:00:00 GMT http://hdl.handle.net/10027/8387 2011-08-05T05:00:00Z