tbuckley/cabot-search huggingface dataset:
Highly Efficient Retroviral Gene Transfer Based on Centrifugation-Mediated Vector Preloading of Tissue Culture Vessels
Authors: Klaus Kühlcke, Boris Fehse, Andrea Schilz, Sonja Loges, Carsten Lindemann, Francis Ayuk, Friederike Lehmann, Norbert Stute, Axel A. Fauser, Axel R. Zander, Hans-Georg Eckert
Journal: Molecular Therapy (2002)

Efficient retroviral gene transfer into primary cells is a prerequisite for various gene therapeutic strategies. We have developed a transduction protocol based on the preloading of tissue culture vessels with retroviral particles by low-speed (1000<i>g</i>) centrifugation. We show that vector-preloaded tissue culture vessels allow highly efficient gene transfer into various target cells. We obtained transduction rates of up to 85% for primary T lymphocytes after just a single round of transduction. Under clinically relevant conditions using a vector developed for suicide gene therapy and produced under good manufacturing practice (GMP) conditions, the described method allowed generation of large numbers (>2×10<sup>9</sup>) of gene-modified T cells. The preloading concept ensures transduction of target cells in their optimal growth medium regardless of the medium used for vector production. This facilitated highly efficient gene transfer into quite different target cells such as CD34<sup>+</sup> and AC133<sup>+</sup> bone marrow progenitor as well as mesenchymal stem cells. The presented method combines high gene-transfer rates with a great potential for standardization in accordance with GMP guidelines and is consequently well suited for both research and clinical applications.
The Aggressiveness and Extent of Meningioma Imaged by Combined Fluorine-18 FDG and Positron Emission Tomography
Authors: C. OLIVER WONG, MAXIMO V. BLEZA, HOWARD J. DWORKIN
Journal: Clinical Nuclear Medicine (2000)

A 52-year-old woman with recurrent left cavernous sinus meningioma had undergone partial resection 5 years earlier. She had progressive symptomatic enlargement of the residual tumor and was considering further resection and radiotherapy. Results of the initial fluorine-18 fluorodeoxyglucose positron emission tomograph (F-18 FDG PET) obtained to judge tumor aggressiveness was negative, suggesting low-grade tumor. Sodium fluoride ion (18F−) was subsequently injected with a dose equal to the calculated residual F-18 FDG in the body. The combined F-18 FDG/18F− scan, performed 1 hour after 18F− ion injection, revealed intense uptake by the deposition of 18F− ion in the calcified tumor against background activity of F-18 FDG. This combined imaging protocol not only ensures exact coregistration of 18F− and F-18 FDG but also increases the sensitivity for detecting recurrent tumor with low metabolism by the ubiquitous presence of calcification in this tumor. The combined protocol would be useful to show the extent and aggressiveness of this disease as it measures tumor metabolism and calcification.
Abstract 4672: Overexpression of NR5A2/LRH1 in pancreatic cancer and possible association with clinical outcome
Authors: Xiaoqun Dong, Yanan Li, William E. Fisher, Ping Chang, James L. Abbruzzese, Donghui Li
Journal: Cancer Research (2011)

Abstract Pancreatic cancer is the 4th leading cause of cancer deaths in the United States and it has the lowest 5-year survival rate in all tumors. Genome-wide association study (GWAS) has identified a significant association between NR5A2/LRH1 gene variants and risk for pancreatic cancer. LRH1, a direct target of PDX1, governs liver and pancreas differentiation in early embryonic development and controls cholesterol/bile-acid homeostasis and steroidogenesis in adulthood. LRH1 promotes cell proliferation by interacting with β-catenin/Tcf4 signaling and upregulating estrogen metabolic genes. We hypothesize that LRH1 acts as an oncogene in pancreatic cancer. We measured LRH1 protein expression by immunohistochemistry (IHC) in 129 resected pancreatic tumors and LRH1 mRNA expression by RT-PCR in 15 frozen paired tumor and adjacent normal tissues. We examined the differences in expression levels between tumor and normal tissues using paired t-test. We evaluated the correlation of protein or mRNA expression levels with overall survival (OS) or tumor characteristics by Cox regression or χ2 test, respectively. IHC showed both nuclear and cytoplasm staining for LRH1. Tumors had a higher level of LRH1 expression than normal tissue did (Mean ± SD of staining score: 5.46 ± 1.64 vs. 3.73 ± 1.77, respectively, p&amp;lt;0.001). Patients with a higher level of nuclear LRH1 expression had worse survival (HR = 2.41, 95% CI=0.97-6.00, p=0.059) but patients with a higher level of cytoplasmic LRH1 expression had better survival (HR = 0.36, 95% CI = 0.09-1.46, p=0.15). Eight out of 8 tumors (100%) with lower nuclear LRH1 expression (staining score &amp;lt;3) were well or moderately differentiated, while 44 out of 121 (36.4%) of tumors with higher nuclear LRH1 expression (staining score ≥ 3) were poorly differentiated (p=0.03). The LRH1 mRNA expression level was 8-fold higher in tumors than in normal tissues (p&amp;lt;0.001). The observed association between LRH1 overexpression and worse clinical outcome and more aggressive tumor type supports the hypothesis that LRH1 acts as an oncogene in pancreatic cancer. Whether LRH1 can be a putative therapeutic target for the treatment of pancreatic cancer needs further investigation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4672. doi:10.1158/1538-7445.AM2011-4672
PTH-077 The Changing Face of Clostridium Difficile Infection: Abstract PTH-077 Table 1
Authors: N M Joshi, J Seale, S Das
Journal: Gut (2013)

<h3>Introduction</h3> C.<i>difficile</i> infection (CDI) is the most common identified cause of antibiotic associated diarrhoea and carries a significant mortality. Several reports have demonstrated that exogenous infection plays an important role in the spread of CDI. Reports show that ribotype 027 has been responsible for large outbreaks of CDI and is associated with a poorer outcome. <h3>Methods</h3> All cases of CDI over a 9 month period (ending August 2012) were cultured and typed by the London reference laboratory. For each case, retrospective data on patient demographics, admission dates, ward and clinical team were analysed. <h3>Results</h3> 32 new cases of CDI occurred of which 22(69%) could be ribotyped. All cases had had antibiotic exposure. Average age: 67 years, 27% of cases were from patients admitted to critical care and 13% were under elderly care. 27% of all cases were community and 73% hospital acquired. 12 ribotypes were seen (table 1), 1 case of type 027. There were no cases of CDI of the same ribotype originating in the same clinical area or under the care of the same clinical team within 30 days of each other. 1 patient (ribotype 015) underwent colectomy for colonic perforation secondary to extensive pseudomembranous colitis with co-existing diverticular disease. There was 16% overall mortality on index admission with 1 death indirectly attributable to CDI (ribotype 020). <h3>Conclusion</h3> In contrast to previous literature, type 027 was not the predominant ribotype seen in our cohort. The case requiring colectomy was type 015 and the death indirectly attributed to CDI was also not caused by type 027. This demonstrates a possible shift in the epidemiology of CDI. The groups most at risk were patients admitted to critical care and those under the care of the elderly care physicians with an overall 16% mortality whilst still admitted. There was little evidence of cross-infection and most cases were endogenously acquired indicating that infection prevention and control methods being practised at our Trust are effective. These findings also suggest that the main cause of CDI in this study arises from selection pressure secondary to antimicrobial use and emphasises the importance of antibiotic stewardship in the prevention and control of this infection. <h3>Disclosure of Interest</h3> None Declared.
Abstract P4-12-04: Association of single-strand breaks (SSBs) in normal breast DNA with estimates of breast cancer risk
Authors: RT Chatterton, M Sahadevan, RE Heinz, S Sukumar, V Stearns, MJ Fackler, O Lee, I Sivaraman, K Kenney, SA Khan
Journal: Cancer Research (2012)

Abstract Background: Formation of single strand breaks in DNA is a constant process, estimated to occur 10,000 times per day in each cell, either from endogenous biosynthetic errors or from interference by endogenous or exogenous agents. Base excision repair in some cases may be impaired or may be exceeded by the rate of DNA damage, leading to cancer. Nick translation of with labeled nucleotides can be used as a quantitative indicator of SSBs. Methods: Healthy women were recruited through the Love-Avon Army of Women and breast clinics of Northwestern and John Hopkins Universities. Digital or digitized mammograms were evaluated for percent density using Cumulus software. The medians (ranges) were age 51 (36 to 60); BMI 28.2 (18.7 to 51.3); life-time Gail estimate 12.5 (5.6 to 28.3); % breast density 16.5 (2.4 to 52.5); Masood score 13 (0 to 18). Breast tissue was obtained by random fine needle aspiration (rFNA). Specimens were rinsed into ice-cold phosphate buffered saline and were stored at −80°C. Thawed samples were centrifuged at 2200 g for 60 min. A kit from Norgen Inc., was used to separate DNA, RNA, and protein from the pellet. The lipids were extracted with ethyl acetate-hexane (3:2) from the supernatant fluid, and triglycerides were precipitated from cold 90% methanol, leaving a purified lipid fraction containing the steroids. Steroids were then fractionated by HPLC on a C18 column, and estradiol was analyzed by a radioimmunoassay. Blood was obtained at the same time as the rFNA samples and was frozen prior to analysis. An aliquot containing 200 ng of DNA (260/280 ratio 1.3 to 2.3) was taken for the nick translation assay. Incorporation of free nucleotides at SSBs with dCTP labeled with3H was catalyzed by Polymerase I from E. coli. A quality control preparation prepared from calf thymus DNA and a reagent blank without DNA was included with each set of 6 samples. Incorporation of3H-dCTP was linear with dose of DNA and reached a maximal at 30 min. Separation of free from incorporated3H-dCTP was accomplished by gel chromatography on an 80 × 15 mm column. The concordance of interassay values was 0.96. Results: Incorporation of nucleotides into DNA ranged from 0.03 to 8.59 pmol/μg, median 0.63 pmol/μg DNA. The association with other factors associated with breast cancer is shown in the Table. A significant correlation, was found with % breast density, life-time risk by the Gail model, but not serum estradiol concentrations. Conclusions: Assessment of SSBs by the nick translation procedure may be a useful indicator of breast cancer risk. Future studies will relate this method with actual risk as assessed by analysis of pre-diagnosis specimens with subsequent occurrence of breast cancer. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-12-04.
Abstract P4-06-15: Mifepristone modifies the tumor microenvironment increasing the therapeutic efficiency of low doses of Doxorubicin liposomes or paclitaxel-albumin nanoparticles in a murine model of breast cancer.
Authors: GR Sequeira, SI Vanzulli, CA Lamb, PA Rojas, C Lanari
Journal: Cancer Research (2012)

Abstract Chemotherapy is the standard treatment for metastatic breast cancer today. To overcome undesired side effects and improve drug efficacy in tumor cells, different nanoparticle formulations have been developed. Paclitaxel has been bound with human albumin (Nab-paclitaxel), and pegylated liposomes have been developed for doxorubicin delivery (PEG-LD). The aim of our study was to evaluate the effect of these therapeutic agents in experimental mammary carcinomas expressing different ratios of progesterone receptor (PR) isoforms and to evaluate the effect of combined treatments of chemotherapeutic agents with the antiprogestin mifepristone (MFP). Nab-paclitaxel (Abraxane, Abraxis), at a dosage of 30 and 60 mg/kg body weight administered in three doses every four days, completely inhibited the growth of the antiprogestin-resistant C4-2-HI mouse mammary carcinoma, which is associated with high levels of PR isoform B (PR-B) expression. C4-HI, an antiprogestin-responsive mouse tumor that shows high levels of PR isoform A (PR-A) expression, was not inhibited by the low Nab-paclitaxel dose. However, the combination of MFP (6 mg silastic pellet) with Nab-paclitaxel improved the efficacy of both single treatments. Similar experiments were conducted using PEG-LD (Doxopeg; Laboratorios Raffo). In this case, all tumors tested (C4-HI, CC4-3-HI, 59–2-HI and C4-2-HI) were all highly responsive to high (18 mg/kg/week) or low (9 mg/kg/week) PEG-LD doses. When PEG-LD concentrations were lowered to 4.5 mg/kg/week and they were combined with MFP, an improved response was observed with combined treatments only in tumors with high PR-A levels (p &amp;lt;0.001). Using BALB/c-GFP mice, we demonstrated that MFP treatment induced tissue remodeling in PR-A positive tumors, increasing the stromal/tumor cell ratio and the number of vessels (as shown by CD31 positive cells; p &amp;lt; 0.001). Taking advantage of doxorubicin's red autofluorescence, we showed increased levels of drug accumulation in tumor cells surrounding the stromal tissue. Using a human albumin antibody we demonstrated nab-paclitaxel accumulation in similar locations. However, because increased areas (p &amp;lt; 0.001) of stromal tissue were observed in the antiprogestin-treated mice, the total amount of PEG-LD or Nab-paclitaxel was much higher in the combined-treated tumors. We propose that antiprogestins have the potential to enhance the efficacy of nanotherapeutics in patients with breast tumors with a high PR-A/PR-B ratio. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-15.
SP0178 Gout and Crystal-Induced Diseases: Year in Review 2013-2014
Authors: F.P. Perez-Ruiz
Journal: Annals of the Rheumatic Diseases (2014)

<h3></h3> In recent times, increasing interest has been raised on crystal-induced arthritis, especially regarding gout and the development of new medications to treat hyperuricaemia and inflammation, what has been nicely demonstrated by bibliometric studies. Nevertheless, pyrophosphate arthritis (PPA) remains as the “orphan crystal disease”. Risk factors for developing PPA have been recently published. In the last year, new mechanistic definitions for hyperurcaemia and gout have been proposed Genomics have shown different polymorphisms of renal and intestinal transporters to be associated with increased risk of hyperuricaemia and gout, the metabolic origin of hyperuricemia (overproduction) seeming to be marginal nowadays. In addition, the impact of environmental factors, such as diet and medications has been also investigated. Imaging is emerging as a useful tool to evaluate the presence of urate deposits and the burden of disease, and the clinical applicability of each of the new imaging techniques, especially ultrasonography and dual energy computed tomography, coming more close to focus. The importance of inadequate or absence of treatment on the appearance of flares during in-hospital admissions has been recently highlighted. To this point, a proof-of-concept educational intervention on patients, by Rees and coworkers at Nottingham University, has shown to be extremely productive regarding to achievement of therapeutic goals. A bunch of new medications targeting hyperuricemia of gout are in the pipeline, and some results became available recently. Others, but restricted to a patient-profiled indications such as pegloticase and canakinumab, have been approved by EMA in the last year, others currently used with either empiric or off-label indications. <h3>Disclosure of Interest</h3> F. Perez-Ruiz Grant/Research support from: Ministerio de Sanidad, Gobierno de España, Asociacion de Reumatόlogos de CRuces, Consultant for: Astra-Zeneca, Menarini, Metabolex, Novartis, Pfizer, SOBI, Conflict with: Menarini, Astra-Zeneca <h3>DOI</h3> 10.1136/annrheumdis-2014-eular.6166
Abstract MS1-2: Long Noncoding RNAs in Breast Cancer Progression
Authors: Chang HY
Journal: Cancer Research (2010)

Abstract Long intergenic noncoding RNAs (lincRNAs) are pervasively transcribed in the genome, yet their potential involvement in human disease is not well understood. Recent studies of dosage compensation, imprinting, and homeotic gene expression suggest that individual lincRNAs can function as the interface between DNA and specific chromatin remodeling activities. We found that many lincRNAs become systematically dysregulated during breast cancer progression. A lincRNA termed HOTAIR is increased in expression in primary breast tumors and metastases, and HOTAIR expression level in primary tumors is a powerful predictor of eventual metastasis and death. Enforced expression of HOTAIR in epithelial cancer cells induced genome-wide re-targeting of Polycomb Repressive Complex 2 (PRC2) to an occupancy pattern more resembling embryonic fibroblasts, leading to altered histone H3 lysine 27 methylation, gene expression, and increased cancer invasiveness and metastasis in a manner dependent on PRC2. Conversely, loss of HOTAIR can inhibit cancer invasiveness, particularly in cells that possess excessive PRC2 activity. A second long noncoding RNA, named PANDA, is part of the DNA damage response mediated by p53 and controls the balance between cell death and cell cycle arrest. Collectively, these findings suggest that lincRNAs play active roles in modulating the cancer epigenome and may be important targets for cancer diagnosis and therapy. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr MS1-2.
SP0055 Licensing New Drugs in SLE: The EMA Draft Guideline
Authors: B. Heelan
Journal: Annals of the Rheumatic Diseases (2013)

<h3></h3> The European Medicines Agency has released the draft Guideline on clinical investigation of medicinal products for the treatment of systemic lupus erythematosus, cutaneous lupus and lupus nephritis. A request for input and comments to this draft guideline by September 2013 is made. Guidelines are intended to provide a basis for practical harmonisation of the manner in which the EU Member States and the European Medicines Agency interpret and apply the detailed requirements for the demonstration of quality, safety and efficacy contained in the Community directives. Guidelines direct drug developers in their clinical programme such that: - Ø The requirements of and the options for how to demonstrate quality safety and clinically relevant efficacy are clear - Ø Provide an EU agreed position on the requirements This presentation will give an overview of the process of guideline development and specifically address the main issues required for the demonstration of efficacy in SLE, cutaneous lupus and lupus nephritis. It is highlighted that in order to demonstrate a reduction disease activity, patients need to have a clinically important and sufficient level of disease activity prior to treatment in order to demonstrate a significant change. The use of composite scores such as BILAG, ECLAM, LAI, SLEDAI and SLAM, ideally a combination of these, is considered appropriate and may be complemented with global patient assessments of the disease with visual analogue scales and health related quality of life. The efficacy aims can be considered as - reduction in disease activity (induction) - reduction in flares (maintenance) - prevention of long-term damage. Efficacy should be demonstrated preferably through validated composite indexes in which the effect seen in an objective measure of reduction in global disease activity is not offset by worsening of the subject's condition overall or worsening in any specific organ system. Additional guidance on cutaneous disease and lupus nephritis as well as endpoints for juvenile SLE are provided. While many external comments received on the original concept paper for SLE released by the EMA have been incorporated, there are limitations in the guidance specifically in relation to certain complications of SLE such as CNS disease. The absence of specific guidance on some features of SLE reflects a lack of validated diagnostic and assessment criteria which could serve as reliable efficacy endpoints. The draft guideline is published on the EMA website for external comments. Once the consultation period ends all external comments received will be reviewed and the draft guideline will be amended as required before release of the final guideline. <h3>Disclosure of Interest</h3> None Declared
1327 STEPWISE COMBINATION OF SIMPLE NON-INVASIVE FIBROSIS SCORING SYSTEMS INCREASES DIAGNOSTIC ACCURACY IN NON-ALCOHOLIC FATTY LIVER DISEASE
Authors: M. Demir, S. Lang, D. Nierhoff, U. Drebber, I. Wedemeyer, T. Goeser, U. Töx, H.-M. Steffen
Journal: Journal of Hepatology (2013)
SP0182 Can We Prevent Bone Loss in Rheumatoid Arthritis by Modern Treatment with Biologics?
Authors: G. Haugeberg
Journal: Annals of the Rheumatic Diseases (2014)

<h3></h3> In this lecture we want to explore the role of inflammation for development of bone damage in rheumatoid arthritis (RA) and illuminate if biologic DMARDs can prevent against RA bone loss mainly focusing on osteoporosis. In RA bone involvement presents as joint erosions and periarticular and generalized osteoporosis. Generalized osteoporosis, an extraarticular manifestation of the RA disease, causes an increased risk of fractures and increases the burden of the RA disease. Inflammation is the driving mechanism behind bone damage in RA. In the absence of synovitis erosions do not occur, further markers of inflammation have been shown to be associated with development of both periarticular and generalized osteoporosis. These bone damage processes starts from the beginning of the disease. To suppress inflammation in RA may not only be important to prevent joint damage but also to avoid generalized inflammatory bone loss and may thereby reduce future fracture risk. The field of osteoimmunology has over the last years increased our knowledge and given us insight into mechanisms involved in the bone damage processes in inflammatory joint disorders. In RA osteoclast recruitment and activation have been shown to be the dominant pathophysiological mechanisms involved in development of erosions and osteoporosis. These processes are driven by macrophage colony stimulating factors (MCSF) and receptor activator of NF-kB ligand (RANKL), and pro-osteoclastiogenic cytokines like e.g. tumor necrosis factor (TNF) and interleukins (IL)- 6 support this process. Targeted anti inflammatory treatment e.g. blocking the effect of TNF and IL-6 is thus a logical treatment principle to prevent bone damage in RA. The importance of RANKL in the development of bone damage including erosions in RA has been shown in a study using the osteoporosis drug denosumab. In this study the inhibition of RANKL by denosumab, a monoclonal antibody against RANKL, was shown to protect against development of erosions in RA, despite ongoing inflammation. From the early beginning of the biologic treatment era in RA it has been known that anti TNF treatment is superior to synthetic DMARDs in reducing radiographic joint damage. Later on other biologic drugs with other modes of action have also shown to reduce the rate of radiographic joint damage (e.g. IL-6 inhibition, T-cell inhibition and B cell depletion). There are also data, however more scarce, that shows that biologic treatment also can reduce the development of generalized bone loss. These data highlights the importance of suppressing inflammatory mechanisms involved in bone damage to maintain bone health in RA to reduce fracture risk. <h3>Disclosure of Interest</h3> None declared <h3>DOI</h3> 10.1136/annrheumdis-2014-eular.6178