tbuckley/cabot-search huggingface dataset:
307. A Mechanism for AZT Mediated Enhancement of Transgene Expression
Authors: Fayaz R. Khazi, Andrew C. Edmondson, Katherine A. High
Journal: Molecular Therapy (2004)

Recombinant adeno-associated viral (rAAV) vectors have been used to treat hemophilia B by efficient transfer of F.IX into liver and muscle in animals; human trials are currently underway. In a recent dose-escalation study investigating muscle directed F.IX gene transfer using rAAV vectors, an HIV-positive subject who received the lowest vector dose showed highest F.IX expression levels. Coadministration of Zidovudine (3'-Azido-3'-deoxythymidine or AZT) during the trial was suggested as an explanation for this observation (Manno et al, Blood 101:8, 2003). AZT is a nucleoside analog that inhibits HIV reverse transcriptase activity. It has also been implicated in inhibiting DNA replication, in generation of oxygen radicals and in induction of transcription factors. We hypothesized a role for the AZT induced nuclear transcription factor, NF-kappaB, in enhancing gene expression by binding to the four consensus NF-kappaB binding sites in the CMV promoter of the vector. Previously, we have shown an AZT dependent increase (30-fold) in expression of a CMV promoter-driven transgene following transduction of HepG2 cells with an AAV vector. Similar experiments using a vector with an EF1alpha promoter showed no change in levels of transgene expression in response to AZT, indicating a CMV promoter-specific response. Western analysis of total cellular extracts from transduced cells cultured for up to 72h in the presence of different levels of AZT showed an increase in NF-kappaB (p65) levels with increasing AZT. NF-kappaB consensus sequence binding assays using nuclear extracts from AZT treated (0.4, 4 and 40mM) cells showed 1.5, 4.1 and 4.3-fold increase, respectively, in NF-kappaB levels over untreated samples. Reporter assays were performed using plasmids with a luciferase gene driven by either a TK (thymidine kinase) minimal promoter or an NF-kB4-TK promoter. Our results show increasing reporter expression (8-fold in 40mM AZT treatment) with increasing concentrations of AZT with NF-kB4-TK promoter, but not with the TK-minimal promoter. We further confirmed our findings by silencing the ATM gene, an activator of NF-kappaB, using siRNA. ATM silenced cells produced significantly lower levels of active NF-kappaB in nuclear extracts of cells treated with AZT. Moreover, the activity of the NF-kB4-TK luciferase promoter decreased 7-fold, indicating a specific role for ATM in CMV driven gene expression via NF-kappaB activation pathway. Our findings thus provide a molecular explanation for enhanced CMV-promoter driven gene expression in a clinical trial subject treated with Zidovudine and provide a novel avenue towards exploring the use of host transcription factors to significantly enhance gene transfer treatment strategies.
Emergency room use of valved holding chambers (VHCs) — Can it be improved?
Authors: Mark Sanders, Ronald Bruin, Cuong Tran, Simon Warren
Journal: European Respiratory Journal (2014)

Background : VHC-use to facilitate pressurised metered dose inhaler (pMDI) drug delivery is recommended in emergencies. Static depletion of the lung dose occurs with first use of VHCs. Anti-static (A-S) measures are not practicable in emergencies (surfactant washing—no time for drying, and priming puffs—wasteful, time-consuming and arbitrary).

Methods : Next Generation Impactor™ (NGI) comparison of delivery of salbutamol sulphate (SS) pMDI via A2A Spacer® VHC (A2A), with A2A used a) detergent-washed and dried per Instructions for Use (IfU), b) unwashed, straight from supplier bag, and c) A-S cloth wiped and A-S bag used during manufacture. SS lung dose was calculated from NGI sum of stages 3-5.

Results : A2A washed and dried per IfU provided the best lung dose, Fine Particle Fraction (FPF) and Fine Particle Dose (FPD) data. The unwashed, ex-bag A2A was poorest, A2A used from the A-S bag gave an intermediate performance, delivering 45% and 66% respectively of the FPD obtained for A2A used per IfU. All paired differences were significant (P<0.05).

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Conclusion : The unwashed, ex-bag data confirm undesirable drug delivery per probable use in emergency situations, and that the adoption of A-S pre-treatment and bagging contributes to improved performance. Optimisation work is being conducted to facilitate use straight from the packaging, without washing or priming the VHC. The data suggest that provision of an A-S pre-treated VHC product is desirable for the emergency room, including ambulance and school use.
Abstract P1-16-06: Margin Probe device is able to reduce re-excision rate of breast conserving surgery in invasive and pre-invasive breast cancer independent from any patient or tumor related factors
Authors: Jens Uwe Blohmer, Julia Tanko, Ragna Voelker, Julia de Grahl, Jessica Gross
Journal: Cancer Research (2015)

Abstract Background: A positive margin status after breast conserving surgery (BCS) is one of the strongest predictors of local recurrence of intraductal and invasive carcinoma. As much as 20-50% of all patients with BCS need to undergo a second operation in order to receive free margins. In this study we tested the clinical performance of Margin Probe (Dune Medical Devices), a novel device for intraoperative margin evaluation. Methods: A prospective clinical trial was performed: The device was applied to 150 lumpectomy specimen from consecutive patients with BCS treated during the first three months in 2013. The re-excision rate was compared to the re-excision rate of a historical group of 156 patients treated with BCS during the first three months in 2012, without the application of the device. We analyzed whether Margin Probe is affected by tumor morphology, grading, size of the tumor, breast density, age, body-mass-index or the use of wire-marker. Results: Due to the application of Margin Probe the re-excision rate decreased significantly by 51% from 39.7% to 14.6%. In the subgroup of intraductal carcinoma (DCIS) the re-excision rate was reduced about two thirds from 66.7% to 23.1%. In the subgroup of invasive lobular carcinomas the re-excision rate decreased from 37.0% to 19.0%. Margin Probe results are not affected by grading, tumor size, breast-density, age, body-mass-index or wire-marker application. Conclusion: Margin Probe is an effective tool for detection of positive margins during BCS and significantly decreases the re-excision rate. It is not limited to invasive carcinoma but also detects involved margins in DCIS as well as in invasive lobular carcinoma. It does not interfere with any of the factors we examined. Citation Format: Jens Uwe Blohmer, Julia Tanko, Ragna Voelker, Julia de Grahl, Jessica Gross. Margin Probe device is able to reduce re-excision rate of breast conserving surgery in invasive and pre-invasive breast cancer independent from any patient or tumor related factors [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-16-06.
Abstract 17205: The Value of QS-Pattern in Leads V1 to V3 for Predicting Improvement of Left Ventricular Function After ST-elevation Myocardial Infarction
Authors: Tetsuo Horimatsu, Kenichi Fujii, Masashi Fukunaga, Takahiro Imanaka, Kojiro Miki, Hiroto Tamaru, Akinori Sumiyoshi, Ten Saita, Machiko Nishimura, Masanori Takada, Yoshiharu Nishibori, Takao Maruyama, Tohru Masuyama
Journal: Circulation (2014)

Background:  Previous studies have reported value of the electrocardiogram (ECG) at admission in predicting infarct size and prognosis after ST-elevation myocardial infarction (STEMI). Despite admitting to the hospital immediately after onset of chest symptoms, some patients’ ECG revealed QS-pattern in anterior chest leads after STEMI. We investigated the impact of QS-pattern on the initial ECG on recovery of left ventricular function in anterior STEMI patients.   Method:  We examined 127 consecutive first anterior STEMI patients with normal preprocedural CK-MB levels who admitted within 6 h from onset of symptoms. All patients underwent primary percutaneous coronary intervention (PCI) for the left anterior descending artery (LAD) lesions immediately after admission. The patients were divided into two groups according to the presence of QS-pattern in V1 to V3 on preprocedural ECG (QS-pattern group: n=30, non-QS-pattern group: n=97). An echocardiogram was obtained within 24 h of presentation and at 3 months after the acute event. A wall motion score (WMS) was calculated using a 16-segment model. Thrombolysis in myocardial infarction (TIMI) grade score at the end of initial procedure was also recorded.   Result:  There were no differences in preprocedural troponin level and time interval from onset to revascularization between two groups. However, the incidence of TIMI grade    Conclusions:  Among patients reached to the hospital soon after the onset of infarction, ECGs revealed QS-pattern in lead V1 to V3 in approximately 20% of patients with first anterior STEMI. These patients showed a greater rise of biomarkers of myocardial necrosis and poor recovery of left ventricular function. This easily assessable ECG pattern is useful in clinical risk stratification for anterior STEMI patients.
In vitro and in vivo characterization of a monoclonal antibody, ATN-658, targeting the uPA system
Authors: Jennifer A. Callahan, Ivy Beck, Todd W. Bauer, Lee M. Ellis, Andrew P. Mazar, Graham C. Parry
Journal: Cancer Research (2005)

6179  A substantial body of in vitro and in vivo data has implicated the urokinase plasminogen activator system, comprised of the serine protease urokinase (uPA), its cell surface receptor uPAR and plasminogen activator inhibitor-1 (PAI-1), in the neo-vascularization, invasion and metastasis of many solid tumors. More recent work has suggested that the interaction of uPAR with other cell surface receptors, particularly integrins, may play a significant role in these processes - independent of uPA catalytic activity. The important role of uPA-uPAR in tumor growth combined with its abundant expression within tumor, but not normal tissue makes it an attractive diagnostic and therapeutic target. We previously described the production and characterization of monoclonal antibodies specific for the D2D3 domain of uPAR. We have extensively characterized one of these antibodies, ATN-658, both in vitro and in vivo. ATN-658 bound immobilized recombinant uPAR and uPAR expressed on the surface of HeLa cells with a KD of ∼ 1 nM. Importantly, ATN-658 bound recombinant suPAR in which all non-essential glycosylation sites were mutated with a similar affinity - indicating that the antibody is specific for the protein portion of uPAR. Interestingly, although [I-125]-labeled ATN-658 bound to HeLa cells with high affinity, its binding was not competed by uPA. Similarly, binding of [I-125]-labeled scuPA to HeLa cells was unaffected by pre-incubation with unlabeled ATN-658. Unlike other antibodies we have tested, ATN-658 was not internalized upon binding to HeLa cells or MDA MB231 cells. To characterize the functional activity of the antibody, migration assays were performed. Although ATN-658 does not disrupt uPA binding to its receptor, ATN-658 inhibited uPA-induced migration of CHO cells expressing uPAR. Similarly, using a modified Boyden chamber model, ATN-658 inhibited the growth factor (HGF and IGF) induced migration of tumor cells. This data suggests that ATN-658 disrupts the interaction of uPAR with other cell surface receptors. The ability of ATN-658 to inhibit primary tumor growth was investigated in a number of in vivo models. To identify an appropriate dose and schedule for antibody treatment the circulating levels of ATN-658 were measured using a modified ELISA assay. Twice weekly treatment IP at 10 mg/kg resulted in a steady state plasma concentration of antibody of ∼0.4 μM, or 100-fold higher than the KD measured by in vitro binding assays. Using this dose and treatment schedule, ATN-658 significantly inhibited tumor growth in an A549 NSCLC early treatment model. Similarly, ATN-658 inhibited tumor growth in a staged A2780 ovarian carcinoma model. Taken together these data suggest that ATN-658, a monoclonal antibody specific for the D2D3 domain of uPAR, significantly inhibits tumor growth by a mechanism independent of uPA binding to its receptor.
Abstract 13304: Insulin-like Growth Factor I (IGF-1) Reduces Lipid Oxidation and Foam Cell Formation via Downregulation of 12/15-lipoxygenase
Authors: Sergiy Sukhanov, Patricia Snarski, Patricia Lobelle-Rich, Charlotte Vaughn, Catherine Kim, Patrick Delafontaine
Journal: Circulation (2014)

We have previously shown that IGF-1 infusion in Apoe-/- mice decreased atherosclerotic plaque size, macrophages (MF) and lipids suggesting that IGF-1 suppressed MF-derived foam cell formation. 12/15-lipoxygenase (LOX) promotes oxidation of low density lipoprotein (LDL) and MF uptake of oxidized LDL (OxLDL) to form foam cells.

We found that IGF-1 infusion (1.5 mg/kg/d) in Apoe-/- mice fed a Western diet decreased plaque LOX expression (immunostaining, 67±6% decrease) and aortic LOX mRNA (36±4% decrease). IGF-1 reduced serum OxLDL (19.7% decrease) and malondialdehyde (a marker of lipid oxidation, 22% decrease), suggesting that IGF-1 suppressed lipid oxidation. To determine the effects of IGF-1 on lipid oxidation and the role of LOX we exposed human THP-1 MF to 0-50 ng/ml IGF-1 for 0-24 h. IGF-1 dose- and time-dependently reduced LOX protein and mRNA levels (49±5% and 48±6% decrease at 24 h, Western blotting and RT-PCR, respectively) and decreased expression of STAT-6 transcription factor (38±4% decrease). STAT6-specific siRNA reduced both STAT6 and LOX protein levels indicating STAT6 involvement in LOX transcriptional regulation. IGF-1-induced LOX decrease was prevented by PI3 kinase inhibition (LY2940032, 50 uM). Incubation of IGF-1-treated THP-1 MF with LDL (150 ug/ml, 48h) showed that IGF-1 markedly suppressed the ability of MF to oxidize lipids (TBARS assay, 65±4% decrease) and to form foam cells (Oil Red staining, 64±4% decrease). Peritoneal MF from LOX-/- mice had a basal reduction in cell-mediated LDL oxidation and foam cell formation vs. wild type (WT) MF. IGF-1 reduced LDL oxidation and formation of foam cells by WT MF but these effects were blocked in LOX-/- MF suggesting that the ability of IGF-1 to reduce LDL oxidation and foam cell formation was dependent on its ability to downregulate LOX.

Thus, IGF-1 reduced LOX expression in atherosclerotic plaques and suppressed lipid oxidation in Apoe-/- mice. IGF-1 decreased LOX expression in cultured MF via a PI3 kinase- and -STAT6 dependent transcriptional mechanism, resulting in marked reduction in MF-mediated LDL oxidation and foam cell formation. These data strongly suggest that IGF-1-induced MF LOX downregulation plays a major role in the ability of IGF-1 to exert anti-atherosclerotic effects.
Abstract 1982: Invading cancer cells are mostly in G0/G1 and resist chemotherapy demonstrated by real-time FUCCI imaging of cell-cycle kinetics in Gelfoam® histoculture
Authors: Shuya Yano, Shinji Miwa, Sumiyuki Mii, Yukihiko Hiroshima, Fuminari Uehara, Hiroyuki Kishimoto, Hiroshi Tazawa, Ming Zhao, Michael Bouvet, Toshiyoshi Fujiwara, Robert M. Hoffman
Journal: Cancer Research (2014)

Abstract Invasive cancer cells are a critical target for chemotherapy in order to prevent metastasis. In the present report, we demonstrate real-time visualization of cell-cycle kinetics of invading cancer in three-dimensional (3D) Gelfoam® histoculture. A fluorescence ubiquitination cell cycle indicator (FUCCI) was used whereby G1/G0 cells express a red fluorescent protein and S/G2/M cells express a green fluorescent protein. With FUCCI imaging, we observed that cancer cells in G0/G1 phase in Gelfoam® histoculture migrated more rapidly and further than the cancer cells in S/G2/M phase. Cancer cells ceased migrating when they entered S/G2/M phases and restarted migrating after division and re-entry into G1/G0. Migrating cancer cells also were resistant to chemotherapy since they were preponderantly in G1/G2 and cytotoxic chemotherapy targets cells in S/G2/M phase. The results of the present report suggest that novel therapy targeting G1/G2 cancer cells is necessary to prevent metastasis. Citation Format: Shuya Yano, Shinji Miwa, Sumiyuki Mii, Yukihiko Hiroshima, Fuminari Uehara, Hiroyuki Kishimoto, Hiroshi Tazawa, Ming Zhao, Michael Bouvet, Toshiyoshi Fujiwara, Robert M. Hoffman. Invading cancer cells are mostly in G0/G1 and resist chemotherapy demonstrated by real-time FUCCI imaging of cell-cycle kinetics in Gelfoam® histoculture. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1982. doi:10.1158/1538-7445.AM2014-1982
Abstract 19811: Circulating Soluble ST2 Levels are Independent of Cardiac Production after Acute Myocardial Injury
Authors: Vincent Y See, Jessica Fugate, Gagandeep Gurm, Dawn Ament, Timm-Michael Dickfeld, Stephen R Shorofsky, Anastasios P Saliaris, Thomas Klein, Benjamin Remo, Robert Christenson, Christopher R deFilippi
Journal: Circulation (2014)

Background: Soluble ST2 (sST2) is a decoy receptor that modulates the anti-hypertrophic and anti-fibrotic IL-33/ST2 pathway. sST2 is associated with adverse prognosis in HF and ACS; both states are associated with cardiac injury. However, sST2 levels are also increased in non-cardiac disease. To determine the extent of cardiac production of sST2 both chronically and after acute myocardial injury, we measured fractional cardiac production of sST2 compared to the cardiac specific injury marker high sensitive (hs) cTnT before and after elective cardiac ablation. Methods: Twenty-three patients undergoing ablation of atrial fibrillation were enrolled [Caucasian 22 (96%); Male 17 (74%); Age 57.7±10.4 years; eGFR 58.7 ± 0.6 mL/min/1.73m2). Of the 23, 15 (65%) had paroxysmal atrial fibrillation. At baseline, 14 (61%) were in sinus rhythm. Samples were obtained from the radial artery and coronary sinus (CS) pre- and post-ablation. sST2 was measured by the Presage assay. Post ablation samples were measured 201±44 minutes after the first ablation. Results: At baseline, in contrast to cTnT, there was no cardiac production of sST2. Both arterial and CS levels of sST2 increased after ablation but not nearly to the extent of cTnT. No net cardiac production of sST2 was noted (Table). Conclusion: Cardiac production of sST2 is not observed at baseline. Systemic sST2 levels modestly increase following acute myocardial injury. Despite direct myocardial injury, no cardiac production of sST2 is observed. The source of sST2 is extra-cardiac both chronically and after acute injury.
Studies on the Effects of Surgery on Transplanted Animal Tumors and Their Metastases
Authors: Ketcham, Alfred S., Wexler, Hilda, Mantel, Nathan
Journal: JNCI Journal of the National Cancer Institute (1961)

Journal Article Studies on the Effects of Surgery on Transplanted Animal Tumors and Their Metastases Get access Alfred S. Ketcham, M.D., Alfred S. Ketcham, M.D. The Surgery Branch and the Biometry Branch, National Cancer Institute,3Bethesda, Maryland Search for other works by this author on: Oxford Academic PubMed Google Scholar Hilda Wexler, M.A., Hilda Wexler, M.A. The Surgery Branch and the Biometry Branch, National Cancer Institute,3Bethesda, Maryland Search for other works by this author on: Oxford Academic PubMed Google Scholar Nathan Mantel, M.A. Nathan Mantel, M.A. The Surgery Branch and the Biometry Branch, National Cancer Institute,3Bethesda, Maryland Search for other works by this author on: Oxford Academic PubMed Google Scholar JNCI: Journal of the National Cancer Institute, Volume 27, Issue 6, December 1961, Pages 1311–1321, https://doi.org/10.1093/jnci/27.6.1311 Published: 01 December 1961 Article history Received: 09 June 1961 Published: 01 December 1961
Administration of Control-Released Hepatocyte Growth Factor Enhances the Efficacy of Skeletal Myoblast Transplantation in Rat Infarcted Hearts by Greatly Increasing Both Quantity and Quality of the Graft
Authors: Keiichi Tambara, Goditha U. Premaratne, Genichi Sakaguchi, Naoki Kanemitsu, Xue Lin, Hiroyuki Nakajima, Yutaka Sakakibara, Yu Kimura, Masaya Yamamoto, Yasuhiko Tabata, Tadashi Ikeda, Masashi Komeda
Journal: Circulation (2005)

We investigated whether simultaneous administration of control-released hepatocyte growth factor (HGF) enhances the efficacy of skeletal myoblast (SM) transplantation (Tx) through its antiapoptotic, angiogenic, and antifibrotic effects in myocardial infarction (MI).Forty-eight Lewis rats with chronic MI were divided into 4 groups. In Group I (n=14), neonatal SMs (5 x 10(6)) were transplanted in the MI area with a gelatin sheet incorporating 40 microg (1 g/L) of HGF applied. Group II (n=14) had SM Tx and placement of a saline sheet. Groups III (n=10) and IV (n=10) had culture medium injection plus HGF and saline sheet application, respectively. Four rats each from Groups I and II were sacrificed at day 1 for TUNEL assay on donor SMs. The percentage of TUNEL-positive donor cells was much lower in Group I than in Group II (P<0.05). At 4 weeks, in Group I, left ventricular diastolic dimension was smallest in echocardiography, end-systolic elastance was highest, and tau was the lowest (both P<0.0005 in ANOVA) in cardiac catheterization. Vascular density inside the graft was higher in Group I than in Group II (P<0.0001). The percentage of fibrotic area inside the graft was smaller in Group I than in Group II (P<0.001). The graft volume as estimated by fast skeletal myosin heavy chain-positive areas was approximately 7-fold larger in Group I than in Group II (P<0.0001).In SM Tx, HGF can greatly increase the graft volume and vascularity and reduce fibrosis inside the graft, which enhances the efficacy of SM Tx to infarcted hearts.